Intact Head Research Articles

The Influence of Three Commercial Soy Lecithin-Based Semen Extenders and Two Spermatozoa Concentrations on the Quality of Pre-Freeze and Post-Thaw Ram Epididymal Spermatozoa.

There are limited studies on the factors affecting the success of ram epididymal spermatozoa (REPS) cryopreservation. On this note, the current study assessed the influence of three commercial soy lecithin-based semen extenders, AndroMed® (AND), BioXcell® (BIO), and OviXcell® (OVI), and two concentrations (400 × 106 vs. 200 × 106 spermatozoa/mL) on the pre-freeze and post-thaw quality of REPS. The REPS were retrieved from nine adult rams' testes and diluted with each of the three extenders to both concentrations. Straws were frozen manually. Standard motility (SMP) and kinematic parameters (KPs) were assessed via a CASA, while spermatozoa viability, morphology, and acrosomal integrity were assessed via the Kovács-Foote staining technique. The concentration did not significantly affect the pre-freeze and post-thaw SMP and KPs of REPS. BIO and OVI had significantly higher pre-freeze and post-thaw BCFs, post-thaw VAP, and the percentage of all intact heads than AND. In contrast, AND had a significantly lower percentage of REPS with tail defects than BIO and OVI. The 400 × 106 spermatozoa/mL concentration resulted in a significantly higher percentage of all intact heads than the 200 × 106 spermatozoa/mL concentration. Freezing significantly increased tail defects and decreased the percentage of REPS with distal cytoplasmic droplets. The cryopreservation of REPS at the 400 × 106 spermatozoa/mL concentration is recommended. All three extenders must be optimized to preserve the viability, membrane integrity, and better normal morphology of REPS; the reason for increased tail abnormality after the freezing/thawing process needs to be studied.

Single‐domain antibody‐based non‐invasive in vivo imaging of α‐synuclein or tau pathology

AbstractBackgroundSynucleinopathies and tauopathies are characterized by progressive deposition of α‐synuclein (α‐syn) and tau proteins, respectively. These diseases have been linked to proteins mutations or excess α‐syn gene, indicating prominent roles of these proteins in their etiology and pathogenesis. Consequently, being able to non‐invasively image these aggregates would allow early and accurate diagnosis to facilitate clinical trials on drugs targeting these lesions and eventually for prophylactic therapies. Recently, several dye‐based imaging probes with selectivity for tau aggregates have been developed but suitable imaging biomarkers for synucleinopathies are still unavailable. However, these β‐sheet binders bind to different amyloids and thereby lack specificity. Compared to β‐sheet dyes, single domain antibodies (sdAbs), found in camelids and a few other species, are highly specific and their small size allows better brain entry and distribution than whole antibodies.MethodTo develop anti‐α‐syn or anti‐tau sdAbs, we immunized llamas with α‐syn and tau preparations, respectively. Phage display libraries were generated from peripheral blood mononuclear cells and screened for binding to α‐syn or tau proteins using various assays, resulting in prototype sdAbs deemed promising for in vivo imaging. Subsequently, these anti‐α‐syn or anti‐tau sdAbs were labeled with a near‐infrared tag and injected intravenously in synucleinopathy, tauopathy or control mice, followed by imaging in an In Vivo Imaging System (IVIS). Subsequently, brains were extracted for tissue analysis.ResultIntravenous injection of labeled anti‐α‐syn or anti‐tau sdAbs resulted in strong in vivo brain signal detected through the intact head in synucleinopathy and tauopathy mice, respectively, but not in control mice. Notably, anti‐α‐syn sdAbs did not give signal in tauopathy mice and vice versa. The in vivo brain signal correlated strongly with insoluble, soluble, and phospho‐antigen within the brain. Postmortem analysis revealed extensive co‐localization of the sdAb imaging probe with α‐syn or tau aggregates within neurons in the endosomal‐lysosomal system, indicating their interaction in these degradation pathways.ConclusionThese anti‐α‐syn or anti‐tau sdAb probes allow non‐invasive and specific in vivo imaging of α‐synuclein vs tau pathology in mice, with brain signals correlating strongly with lesion burden. These small antibody derivatives have great potential for in vivo diagnosis of these diseases.

Head and body cues guide eye movements and facilitate target search in real-world videos.

Static gaze cues presented in central vision result in observer shifts of covert attention and eye movements, and benefits in perceptual performance in the detection of simple targets. Less is known about how dynamic gazer behaviors with head and body motion influence search eye movements and performance in perceptual tasks in real-world scenes. Participants searched for a target person (yes/no task, 50% presence), whereas watching videos of one to three gazers looking at a designated person (50% valid gaze cue, looking at the target). To assess the contributions of different body parts, we digitally erase parts of the gazers in the videos to create three different body parts/whole conditions for gazers: floating heads (only head movements), headless bodies (only lower body movements), and the baseline condition with intact head and body. We show that valid dynamic gaze cues guided participants' eye movements (up to 3 fixations) closer to the target, speeded the time to foveate the target, reduced fixations to the gazers, and improved target detection. The effect of gaze cues in guiding eye movements to the search target was the smallest when the gazer's head was removed from the videos. To assess the inherent information about gaze goal location for each body parts/whole condition, we collected perceptual judgments estimating gaze goals by a separate group of observers with unlimited time. Observers' perceptual judgments showed larger estimate errors when the gazer's head was removed. This suggests that the reduced eye movement guidance from lower body cueing is related to observers' difficulty extracting gaze information without the presence of the head. Together, the study extends previous work by evaluating the impact of dynamic gazer behaviors on search with videos of real-world cluttered scenes.

Single domain antibody‐based non‐invasive in vivo imaging of tau pathology

AbstractBackgroundNumerous studies have shown that the extent of pathological tau protein in the brain correlates strongly with the severity of Alzheimer’s dementia at the time of death, and similar association with functional phenotypes have been seen in other tauopathies. In recent years, several dye‐based imaging probes with selectivity for tau aggregates have been developed. However, all are β‐sheet binders with varying affinity for different amyloids, and thereby lack specificity for tau lesions. In contrast, single domain antibodies (sdAbs) are specific for their target and because of their small size have much greater access into the brain than standard whole antibodies.MethodTo examine sdAbs’ suitability as non‐invasive in vivo imaging probes for tauopathies, we immunized a llama with a full‐length recombinant tau protein followed by boosters with human brain‐derived pathological tau proteins. Phage display libraries were generated from its peripheral blood mononuclear cells and screened for binding to tau proteins using various assays, resulting in a prototype sdAb deemed promising for in vivo imaging. Subsequently, this anti‐tau sdAb was labeled with a near‐infrared tag (680 nm), and injected intravenously in tauopathy and control mice, followed by imaging using an In Vivo Imaging System (IVIS). After the imaging, brains were extracted for tissue analysis.ResultIntravenous injection of labeled sdAb resulted in a strong in vivo brain signal detected through the intact head in two different models of transgenic tauopathy mice, but not in wild‐type or transgenic α‐synucleinopathy mice. Importantly, the in vivo brain signal correlated strongly with insoluble (r = 0.9839, p < 0.0001) and soluble (human tau (CP27) and phospho‐tau (PHF1): r = 0.9603 and 0.9459, p < 0.0001) tau protein within the brain. Furthermore, postmortem analysis revealed extensive co‐localization of the sdAb imaging probe with tau aggregates within neurons in the endosomal‐lysosomal system, indicating their interaction in these degradation pathways.ConclusionOverall, this specific sdAb imaging ligand has great potential as an in vivo diagnostic marker for Alzheimer's disease and related tauopathies, and sdAb‐based imaging may be applicable to a variety of protein conformational disorders.

Blunt force trauma in the human mandible: An experimental investigation

There exist sparse data associating known points of mandibular impact with resultant fracture patterns. These data may provide an important foundation for interpreting mandible fractures in forensic cases. The current study illustrates results of experimental blunt force impacts to five locations on the mandible: the ramus, posterior body, mid-body, anterior body, and midline. The experimental sample comprised thirteen intact heads from non-edentulous, unembalmed human adult male cadavers. Three key findings resulted from this study: (1) each impact produced a fracture at or adjacent to the impact site; (2) ramus and midline impacts produced more fractures than mandibular body impacts; and (3) fracture locations varied among mandibles impacted at the same location and exhibited similarities among mandibles impacted at different locations, suggesting there are limitations in estimating mandibular impact sites based on fracture location alone. Overall, these results contribute a comparative sample of known blunt force trauma cases for practitioners evaluating mandibular fractures in forensic cases.

Quantification of the effectivity of laser therapy shortly following brain injury via dual-wavelength laser imaging

Treatment of traumatic brain injury, within a few minutes and even before transportation to the nearest medical facility, can significantly curtail injury severity and even prevent death. The primary aim of this study was to evaluate the effect of laser irradiation as a therapeutic intervention tool immediately following brain injury. To this end, a dual-wavelength laser speckle contrast imaging (DW-LSCI) system based on two laser sources at two wavelengths 532 and 660 nm was employed to monitor changes in cerebral blood flow, tissue saturation and rate of oxygen consumption in a mouse model of intact head injury. In addition, structural changes of tissue were evaluated using linear approximation to Rayleigh-MIE scattering in the range between the two laser wavelengths. Furthermore, cerebral tissue temperature was imaged by a thermal camera providing additional information on physiological brain tissue condition. Experiments were conducted on anesthetized mice (n = 6, female) subjected to a closed head weight-drop model of focal brain injury. After 5 min of baseline measurement, focal brain injury was induced and measurements were conducted for 10 min. Low-level laser therapy (LLLT) was than administrated for a duration of 15 min with uniform exposure of 45 J/cm2 from a CW diode laser source (810 nm). Concurrently, measurements were carried out over the treatment time interval. Laser illumination was then blocked and measurements continued for another 20 min, followed by euthanasia. In comparison to baseline measurements, noticeable variations were revealed post-injury which indicate the severity of brain damage. The use of LLLT inhibited the development of complications in the injured mice by increasing blood flow and saturation and overall oxygen consumption level over the injured area which highlights its effectiveness as a neuroprotective agent immediately following brain injury. Different doses of low-level laser irradiation were also tested (n = 12) with less effectiveness on cerebral parameters. The results presented here support our hypothesis that a high dose of laser irradiation as a first aid can attenuate the injury and save the brain from further worse outcome. To the best of our knowledge, the implementation of the DW-LSCI system to monitor brain hemodynamic and metabolic response to LLLT shortly after head injury in intact mouse brain has not been previously reported.

Peptide bioregulator amprost® in the correction of pathospermia in various types of secretory infertility

The aim of the study was to evaluate the clinical and laboratory effectiveness of the peptide bioregulator Samprost® in the correction of pathospermia in men with secretory type of infertility caused by chronic nonbacterial prostatitis (CNP) and varicocele in the postoperative period.Materials and methods. The multicenter study included 30 patients between 23 and 51 years of age: 15 patients with CNP (1st group) and 15 men with infertility due to varicocele in the postoperative period (2nd group). Patients of all groups received Samprost® for 20 days in the form of intramuscular injections of 5 mg daily. The study design provided for a traditional clinical, laboratory and specialized examination with the study of the level of antisperm antibodies, the study of sperm DNA fragmentation by the TUNEL method, the assessment of oxidative stress, a cryotolerance test, and an electron microscopic examination of the ejaculate. Statistical processing of the material was carried out using the paired Student's t test for dependent samples, and the Wilcoxon nonparametric T test was also used. The calculations were performed using the SigmaPlot 12.5 program.Results. The average age of the study participants was 36.91 ± 4.5 years (23-51 years). Among the concomitant and past diseases, the most common were patients with recurrent varicocele, May-Turner syndrome, varicose veins of the pelvis. When evaluating the effectiveness, the results of the study showed a more significant effect of the drug Samprost® in the group of patients with CNP. This concerned an increase in ejaculate volume by 11.5 %, an increase in sperm concentration by 19.4 %. Progressive sperm motility increased more in the group of patients with varicocele by 12.2 %, compared with the CNP group - 11.4 %. Similar data were obtained during cryotest.After 20-day use of the Samprost® a significant decrease in the level of free radicals in the ejaculate was noted: by 2.5 times in the group with CNP and by 2.3 in the group of patients with varicocele. Analysis of these indicators may indicate an antioxidant effect on the effect on the sperm cell, which is under serious stress in the fight against free radicals. This assumption is also confirmed by the EMIS data, which showed a positive dynamics in the number of intact heads of normal form with a normal structure of chromatin and acrosome, the number of spermatozoa with insufficiently condensed immature chromatin.Conclusion. Samprost® is an effective and safe drug for patients with secretory type of male infertility. Based on the methods of statistical analysis, including statistically significant differences in the study groups, Samprost® is more effective in patients with secretory type of male infertility in the case of CNP than in patients with varicocele in the postoperative period. More significant changes relate to the volume, concentration of sperm viability and a decrease in the level of free radicalsThe 20-day course of Samprost® increases progressive sperm motility during cryotest in patients with varicocele and CNP and can be used for substantiated preparation of patients for gamete cryopreservation and its use in preparatory assisted reproductive technologies programs using intra cytoplasmic sperm injection. A decrease in DNA fragmentation and the received electron microscopic examination of the sperm data indicate a possible influence of Samprost® on sperm morphology and require further research in this area.

Review of Whole Head Experimental Cochlear Promontory Vibration with Bone Conduction Stimulation and Investigation of Experimental Setup Effects.

Bone conduction sound transmission in humans has been extensively studied using cochlear promontory vibrations. These studies use vibration data collected from measurements in live humans, whole cadavers, and severed cadaver heads, with stimulation applied either at an implant in the skull bone or directly on the skin. Experimental protocols, methods, and preparation of cadavers or cadaver heads vary among the studies, and it is currently unknown to what extent the aforementioned variables affect the outcome of those studies. The current study has two aims. The first aim is to review and compare available experimental data and assess the effects of the experimental protocol and methods. The second aim is to investigate similarities and differences found between the experimental studies based on simulations in a finite element model, the LiUHead. With implant stimulation, the average cochlear promontory vibration levels were within 10 dB, independent of the experimental setup and preparations of the cadavers or cadaver heads. With on-skin stimulation, the results were consistent between cadaver heads and living humans. Partial or complete replacement of the brain with air does not affect the cochlear promontory vibration, whereas replacing the brain with liquid reduces the vibration level by up to 5 dB. An intact head–neck connection affects the vibration of the head at frequencies below 300–400 Hz with a significant vibration reduction at frequencies below 200 Hz. Removing all soft tissue, brain tissue, and intracranial fluid from the head increases the overall cochlear promontory vibration level by around 5 dB.

PSV-39 The relationship between structural integrity and sperm survival during hypothermic storage of stallion sperm

Abstract The aim of this work was to study the relationship between ultrastructural integrity and sperm survival during hypothermic storage of chilled and cryopreserved stallion sperm. Semen from 37 stallions, were collected during the breeding season. The structural integrity of sperm was studied using a Hitachi 700 electron microscope. Progressive motility was evaluated using an Olympus BX 41 phase contrast microscope. Sperm survival was defined as the ability of spermatozoa to maintain progressive motility above 5% during hypothermic (+4C) storage of sperm. Spearman nonparametric correlation analysis was used. Sperm survival at a temperature +4°C in chilled semen averaged 115.4±11.47 hours. After freezing sperm survival decreased on average to 48.9±5.56 hours (P < 0.001). A reliable correlation between the number of sperm with intact heads and sperm survival at a temperature of + 4°C in diluted (r=0.60; P < 0.001) and thawed (r=0.63; P < 0.001) sperm was established. A significant negative correlation between the number of spermatozoa with degradation of the acrosome in native sperm and their survival (r=-0.48; P < 0.01) under the hypothermic storage was found. For cryopreserved semen, a similar dependence has not been established. A significant correlation between the number of spermatozoa with the normal position of the acrosome and the sperm survival rate in native sperm was also established (r=0.52; P < 0.01). A significant negative correlation between sperm with acrosome hypoplasia and sperm survival was found (r=-0.67; P < 0.001). We assume that the structural integrity and ability of spermatozoa to maintain progressive motility for a long time under conditions of hypothermic storage depends on the quality of spermatogenesis and the resistance of cells to low and ultra-low temperatures. Authors acknowledge financial support from Russian Science Foundation, Grant No: 20-16-00101, development program of Bioresource collections “Cryobank of genetic recourses the All-Russian Research Institute for Horse Breeding.”

Visualization of the bullfrog middle ear

The structure and function of the anuran middle ear has been explored both physiologically and structurally. It is similar to that of some other vertebrates (birds and reptiles) in that it has a two-ossicle system consisting of an extracolumella and a columella. However, there are notable species and gender variations. Peter Narins has made significant contributions to our understanding of this unusual middle ear. One example is that the middle ear of terrestrial frogs has a cartilaginous operculum adjacent to the columellar footplate. Following on Peter's work, to better understand the middle ear in situ, we obtained Micro CT and Ultra High Resolution Helical (UHR CT) scans of the middle ear in intact heads of the bullfrog (Rana catesbeiana) on a Zeiss Xradia 520 Versa and a Siemens Volume Zoom at 10–100 μm isotropic voxel resolutions. Image analyses showed the core of the columella in this species is not a simple column but a twisted shaft. Further, the extracolumella is not constructed of dense cartilage but rather is a complex with exceptional soft, semi-cartilaginous tissues. Implications of these findings for middle ear function will be discussed.

The development of a rapid and reliable method (SAPSWash): For the extraction and recovery of spermatozoa from superabsorbent polymer containing products

This work presents a rapid and reliable method to recover spermatozoa from Super Absorbent Polymers (SAPs) commonly found in sanitary protection products such as nappies and sanitary towels. The use of salt solutions was investigated and a protocol was developed using a calcium chloride (CaCl2) solution to release semen deposited onto a selected SAP containing product. The method was tested on ultra-sanitary towel samples treated with a known amount of semen. A range of treatments were examined; some samples were prepared and immediately frozen for storage and others were allowed to air dry overnight to replicate the condition of similar items recovered for examination in sexual offence cases. The method allowed the collection of low yields of spermatozoa, but these were still sufficient for microscopic identification of intact heads and to obtain ESI17 DNA profiles from all the samples. This report presents the method, the results obtained and discusses prospective adaptations to the method for validation to implement the method into forensic casework.

Feasibility of a label-free X-ray method to estimate brain amyloid load in small animals

BackgroundAmyloid plaque in the brain is associated with a wide range of neurodegenerative diseases such as Alzheimer's and Parkinson's and defined as aggregates of amyloid fibrils rich in β-sheet structures. New methodWe report a label-free method based on small-angle X-ray scattering (SAXS) to estimate amyloid load in an intact mouse head with skull. The method is based on recording and analyzing the X rays elastically scattered from the β-sheets of amyloid plaques in a mouse head at angles smaller than 10° and energies between 30 and 45 keV. The method is demonstrated by acquiring the spectral SAXS data of an amyloid model and an excised head from a wild-type mouse for 600 s. ResultsWe captured the distinct scattering peaks of the amyloid plaques at momentum transfer (q) of 6 and 13 nm−1 associated with β-sheet structure. We first show linear correlation between the mass fraction of the amyloid target and the area under the peak (AUP) of the scattering curve. We report results for estimating amyloid load in a fixed mouse head by recovering the characteristic scattering signal from the amyloid target situated at various locations. The coefficient of variation in the amyloid load estimate is found to be less than 10%. Comparison with existing methodsThere are no previously described label-free X-ray methods for the estimation of amyloid load in an intact head. ConclusionsWe demonstrated the feasibility of a label-free method based on SAXS to potentially estimate brain amyloid in small animals.

Closed Transverse Pinning for Reduction and Fixation of Metatarsal Neck Fractures: Surgical Technique.

Most metatarsal neck fractures can be successfully treated non-operatively in a cast boot. Displaced metatarsal neck fractures tend to be less stable and have a propensity for the distal fragment to angulate, secondary to the strong flexor tendons, which often forces the distal fracture fragment in a plantar direction and leads to relative metatarsal shortening. Most literature is focussed on antegrade fixation of metatarsal neck fractures using pre-bent K wires or thin elastic nails. Apart from the technical challenges, this technique is limited when bones are osteoporotic as the pre-bent distal end of the K-wire may penetrate the plantar cortex of the proximal metatarsal and prevent the wire from entering the medullary canal of the metatarsal and advancing to the fracture site. Furthermore, when the medullary canal is narrow especially in Asian patients, it may be difficult to pass a bent K-wire through the isthmus of the metatarsal shaft. We describe an innovative technique of closed transverse wiring of the metatarsal head necks that has a distinct advantage in Asian population with osteoporotic bones. With percutaneous manipulation using digital pressure, closed reduction of fracture fragments of the most displaced fracture is done under fluoroscopic guidance to achieve a satisfactory alignment followed by closed transverse wiring of the metatarsal heads. With this procedure, adjacent fractures remain stable within an acceptable range because of intermetatarsal ligaments connected to the adjacent intact head. Our technique has a relatively short operating time and allows for early motion of the metatarso-phalangeal joint. This is especially useful for those with osteoporosis, narrow canal, soft tissue compromise, intra-operative failure of ante-grade pinning and in scenarios of limited surgical equipment/expertise.

Anatomical study in exposed area and operating space of extended endonasal approach to the ventral region of cranio-cervical junction

Objective To explore the scope, feasibility and indications of extended neuroendoscopic endonasal approach to reveal the region of cranial-cervical junction. Methods Ten (20 sides) intact frozen adult cadaveric heads were used to simulate the extended neuroendoscopic endonasal approach to remove the anterior tubercle of atlas, part of the atlas anterior arch, odontoid and part of the slope. In the cranial-cervical junction area, we observed the exposed range and measured the distance from the nostril to the anterior tubercle of atlas, the height and width of the anterior arch of the atlas, the distance of bilateral medial margin of occipital condyle, the distance of bilateral medial margin of the foramen lacerum, the vertical distance from the occipital condyle to the homolateral foramen lacerum, and the height, anteroposterior diameter and transverse diameter of the odontoid. Results The exposed range of the extended neuroendoscopic endonasal approach was as follows: when the lesion was located at the foramen magnum, the medial margin of the occipital condyle was the boundary of both sides, the root of the odontoid was the lower boundary, and the lower edge of the occipital bone was the upper boundary. When the lesion was located at the region of cranial-cervical junction, the upper edge of the foramen lacerum on both sides was the upper boundary, and the foramen lacerum and the outer edge of the occipital condyle were the boundaries of both sides of the bone window. By measurement, the distance from the nostril to the anterior tubercle of atlas was 8.9-9.7 cm, with an average of 9.3±0.3 cm. The height of the anterior arch of the atlas was 2.3-3.0 cm, with an average of 2.7±0.3 cm; the width was 1.5-2.0 cm, with an average of 1.8±0.2 cm; the distance of bilateral medial margin of occipital condyle was 1.5-2.0 cm, with an average of 1.8±0.2 cm; the distance of bilateral medial margin of the foramen lacerum was 2.1-2.9 cm, with an average of 2.4 ±0.3 cm; the vertical distance from the occipital condyle to the homolateral foramen lacerum was 1.6-2.2 cm, with an average of 1.9±0.3 cm; the height of the odontoid was 0.9-1.3 cm, with an average of 1.1±0.2 cm, its length was 0.8-1.3 cm, with an average of 1.1±0.2 cm; the transverse diameter of the dentate was 0.9-1.2 cm, with an average of 1.0±0.1 cm.The maximum exposed area of cranio-cervical junction was 6.4-9.8 cm2, with an average of 8.6±1.6 cm2. Conclusion The extended neuroendoscopic endonasal approach with removal of the anterior tubercle of atlas and part of the atlas anterior arch could effectively reveal the cranial-cervical junction area which is suitable for the treatment of odontoid deformity, skull base depression and atlantoaxial joint dislocation as well as resection of epidural tumors in the cranial-cervical junction and ventral and ventral midline tumors. Key words: Natural orifice endoscopic surgery; Anatomy; Cranio-cervical junction; Extended endonasal approach

Identification of putative odorant binding proteins in the peach fruit borer Carposina sasakii Matsumura (Lepidoptera: Carposinidae) by transcriptome analysis and their expression profile

The peach fruit borers Carposina sasakii Matsumura (Lepidoptera: Carposinidae) are serious fruit borers in East Asia. Odorants have been reported to elicit their olfactory responses, including sex pheromone and host plant volatiles. However, little is known about their molecular aspects of olfactory perception to these odorants. Odorant binding proteins (OBPs) are believed to play an important role in insect olfaction, which recognize and bind odorants, and transport them to odorant receptors located on olfactory neurons. We identified 12 putative OBPs (CsasOBP1-12) expressed in intact heads (with antennae and mouthparts) of C. sasakii adult males and females by transcriptome analysis, including 4 putative pheromone binding proteins (PBPs) (CsasOBP1-2, 7, 11) and 4 putative general odorant binding proteins (GOBPs) (CsasOBP6, 9–10, 12). A phylogenetic tree was constructed to characterize these OBPs. An analysis using fluorescence quantitative PCR showed that CsasOBP3, 5, 7, 9–12 have the highest expression level in the intact heads, suggesting that among them there may be the OBPs playing key roles in C. sasakii olfaction. In the 7 OBPs, CsasOBP5, 7, 11–12 have higher level of expressions in intact heads of C. sasakii adult males than females, while CsasOBP3, 9–10 are expressed equally in the two sexes. Moreover, CsasOBP2 is expressed dominantly in the intact heads and wings, with an equal level between them. CsasOBP8 expression is highest in the wings, while CsasOBP1, 4, 6 are expressed dominantly in the abdomens. Our study is helpful for understanding C. sasakii OBPs’ functions and C. sasakii olfaction from molecular view.

Three-dimensional MRI Analysis of Femoral Head Remodeling After Reduction in Patients With Developmental Dysplasia of the Hip.

Previous reports on patients with developmental dysplasia of the hip (DDH) showed that the prereduced femoral head was notably smaller and more nonspherical than the intact head, with growth failure observed at the proximal posteromedial area. We evaluated the shape of the femoral head cartilage in patients with DDH before and after reduction, with size and sphericity assessed using 3-dimensional (3D) magnetic resonance imaging (MRI). We studied 10 patients with unilateral DDH (all female) who underwent closed reduction. Patients with avascular necrosis of the femoral head on the plain radiograph 1 year after reduction were excluded. 3D MRI was performed before reduction and after reduction, at 2 years of age. 3D-image analysis software was used to reconstruct the multiplanes. After setting the axial, coronal, and sagittal planes in the software (based on the femoral shaft and neck axes), the smallest sphere that included the femoral head cartilage was drawn, the diameter was measured, and the center of the sphere was defined as the femoral head center. We measured the distance between the center and cartilage surface every 30 degrees on the 3 reconstructed planes. Sphericity of the femoral head was calculated using a ratio (the distance divided by each radius) and compared between prereduction and postreduction. The mean patient age was 7±3 and 26±3 months at the first and second MRI, respectively. The mean duration between the reduction and second MRI was 18±3 months. The femoral head diameter was 26.7±1.5 and 26.0±1.6 mm on the diseased and intact sides, respectively (P=0.069). The ratios of the posteromedial area on the axial plane and the proximoposterior area on the sagittal plane after reduction were significantly larger than before reduction (P<0.01). We demonstrated that the size of the reduced femoral head was nearly equal to that of the intact femoral head and that the growth failure area of the head before reduction, in the proximal posteromedial area, was remodeled after reduction. Level IV-case series.

Three-dimensional mammalian tooth development using diceCT

ObjectiveThis study aims to develop the Diffusible Iodine-based Contrast-Enhanced CT (diceCT) method for non-destructive imaging of both soft and mineralised tissues. We sought to document the 3D spatio-temporal pattern of mammalian tooth development including multiple tooth classes and generations, using the tammar wallaby (Macropus eugenii) as a model species. DesignWe took microCT scans of developing fetuses and pouch young stained using Lugol’s Iodine (I2KI) contrast agent. Stained versus unstained specimen comparisons were then made to investigate whether staining had improved visualisation of structures. Scan slices were compared to histological sections to confirm the identity of tissues and structures. Tissue layers were digitally segmented to create 3D models. ResultsDiceCT dramatically enhanced visual contrast of soft tissues, allowing differentiation between epithelial and mesenchymal layers. Subvolume scans at higher magnification achieved single-cell layer resolution within relatively large intact heads. We observed in-situ initiating teeth, which progressed through major stages of tooth development including morphogenesis and mineralisation. In addition, we traced the development of other mineralized and unmineralised tissues, such as the cranial bones and the brain, eye and olfactory system. ConclusionsDiceCT was time- and cost-effective in producing complex 3D models of the entire dentition of the tammar wallaby at each developmental stage with tissue-level resolution. The 3D view of soft and mineralised tooth structures allowed us to define tooth class and generation from a developmental perspective. Additionally, the development of other organs can also be documented using the same scans, demonstrating the efficiency and versatility of this technique.

Role of age and injury mechanism on cervical spine injury tolerance from head contact loading

ABSTRACTObjective: The objective of this study was to determine the influence of age and injury mechanism on cervical spine tolerance to injury from head contact loading using survival analysis.Methods: This study analyzed data from previously conducted experiments using post mortem human subjects (PMHS). Group A tests used the upright intact head–cervical column experimental model. The inferior end of the specimen was fixed, the head was balanced by a mechanical system, and natural lordosis was removed. Specimens were placed on a testing device via a load cell. The piston applied loading at the vertex region. Spinal injuries were identified using medical images. Group B tests used the inverted head–cervical column experimental model. In one study, head–T1 specimens were fixed distally, and C7–T1 joints were oriented anteriorly, preserving lordosis. Torso mass of 16 kg was added to the specimen. In another inverted head–cervical column study, occiput–T2 columns were obtained, an artificial head was attached, T1–T2 was fixed, C4–C5 disc was maintained horizontal in the lordosis posture, and C7–T1 was unconstrained. The specimens were attached to the drop test carriage carrying a torso mass of 15 kg. A load cell at the inferior end measured neck loads in both studies. Axial neck force and age were used as the primary response variable and covariate to derive injury probability curves using survival analysis.Results: Group A tests showed that age is a significant (P < .05) and negative covariate; that is, increasing age resulted in decreasing force for the same risk. Injuries were mainly vertebral body fractures and concentrated at one level, mid-to-lower cervical spine, and were attributed to compression-related mechanisms. However, age was not a significant covariate for the combined data from group B tests. Both group B tests produced many soft tissue injuries, at all levels, from C1 to T1. The injury mechanism was attributed to mainly extension. Multiple and noncontiguous injuries occurred. Injury probability curves, ±95% confidence intervals, and normalized confidence interval sizes representing the quality of the mean curve are given for different data sets.Conclusions: For compression-related injuries, specimen age should be used as a covariate or individual specimen data may be prescaled to derive risk curves. For distraction- or extension-related injuries, however, specimen age need not be used as a covariate in the statistical analysis. The findings from these tests and survival analysis indicate that the age factor modulates human cervical spine tolerance to impact injury.

Surgical Landmark of Mandible Angle in the Identification of Cervical Spinal Level.

Mandible angle is considered to be a consistently palpable external landmark for the identification of cervical spinal level by a radiographic study. But this anatomical study aimed to determine the positional relationship between mandible angle and cervical spinal level in cadavers. In this study, the cervical spine of 10 adult cadavers with intact head and neck structure, including 6 males and 4 females, was dissected, and the position of mandible angle (MA) relative to the corresponding cervical spinal level was measured when the head was fixed in the flexion, anatomy position, and extension. The difference between the genders and the sides was analyzed. On the basis of the study of the corpse samples, the reference level of cervical spine was approximated to C2/3 intervertebral disc. The result has confirmed MA as a consistent and convenient landmark in the identification of cervical spinal level. Given some external landmarks do not consistently correspond to the exact level of the cervical spine, MA provides the relatively consistent reference point: C2/3 intervertebral disc. The authors hold that MA is a superior external landmark, which can help surgeons to localize the skin incision before anterior cervical spine surgery.

Evaluation of brainstem disruption following penetrating captive-bolt shot in isolated cattle heads: comparison of traditional and alternative shot-placement landmarks

AbstractCurrently recommended landmarks for captive-bolt euthanasia of cattle often result in failure to penetrate the brainstem. The purpose of this study was to evaluate the ability to disrupt the brainstem by placing the shot at a higher position on the head. Intact heads from euthanased animals or natural mortalities were used for this study. Heads were grouped as adult (&gt; 2 years), young (6-24 months) and neonate (&lt; 1 month) and randomly assigned to either the LOW group (the intersection of two lines drawn from the medial canthus to the top of the opposite ear) or the HIGH group (midline halfway between the top of the poll and an imaginary line connecting each lateral canthus). Each head received a single shot from a CASH penetrating captive bolt with bolt length and power load selected based on manufacturer's recommendations. Computed tomography images of each head were evaluated independently by two veterinary radiologists. Brainstem disruption was assumed to occur if the bolt passed caudal to the presphenoid bone and deep to the third ventricle and was within 1.5 cm of midline. Brainstem disruption occurred in 16/18 adult HIGH and 7/14 adult LOW heads, 13/16 young HIGH and 11/19 young LOW heads, and 11/11 neonate HIGH and 14/14 neonate LOW heads. The higher shot location landmarks used in this study increased the probability of disrupting the brainstem when adult cattle were shot with a penetrating captive bolt which should reduce the risk of regaining sensibility. Reliable brainstem disruption is a precondition for considering penetrating captive bolt as a single-step euthanasia method. Further research is needed to determine if this method will reliably ensure a humane death.