Aided Cortical Auditory Evoked Potentials in Infants With Frequency-Specific Synthetic Speech Stimuli: Sensitivity, Repeatability, and Feasibility.

OBJECTIVES: The cortical auditory evoked potential (CAEP) test is a candidate for supplementing clinical practice for infant hearing aid users and others who are not developmentally ready for behavioral testing. Sensitivity of the test for given sensation levels (SLs) has been reported to some degree, but further data are needed from large numbers of infants within the target age range, including repeat data where CAEPs were not detected initially. This study aims to assess sensitivity, repeatability, acceptability, and feasibility of CAEPs as a clinical measure of aided audibility in infants. DESIGN: One hundred and three infant hearing aid users were recruited from 53 pediatric audiology centers across the UK. Infants underwent aided CAEP testing at age 3 to 7 months to a mid-frequency (MF) and (mid-)high-frequency (HF) synthetic speech stimulus. CAEP testing was repeated within 7 days. When developmentally ready (aged 7-21 months), the infants underwent aided behavioral hearing testing using the same stimuli, to estimate the decibel (dB) SL (i.e., level above threshold) of those stimuli when presented at the CAEP test sessions. Percentage of CAEP detections for different dB SLs are reported using an objective detection method (Hotellings T 2 ). Acceptability was assessed using caregiver interviews and a questionnaire, and feasibility by recording test duration and completion rate. RESULTS: The overall sensitivity for a single CAEP test when the stimuli were ≥0 dB SL (i.e., audible) was 70% for the MF stimulus and 54% for the HF stimulus. After repeat testing, this increased to 84% and 72%, respectively. For SL >10 dB, the respective MF and HF test sensitivities were 80% and 60% for a single test, increasing to 94% and 79% for the two tests combined. Clinical feasibility was demonstrated by an excellent >99% completion rate, and acceptable median test duration of 24 minutes, including preparation time. Caregivers reported overall positive experiences of the test. CONCLUSIONS: By addressing the clinical need to provide data in the target age group at different SLs, we have demonstrated that aided CAEP testing can supplement existing clinical practice when infants with hearing loss are not developmentally ready for traditional behavioral assessment. Repeat testing is valuable to increase test sensitivity. For clinical application, it is important to be aware of CAEP response variability in this age group.

Gross, computed tomographic, and endoscopic anatomy of the equine carpal extensor tendon sheaths.

OBJECTIVE: (1) To describe the computed tomography (CT) and gross anatomy of the equine extensor carpi radialis sheath (ECRS) and common digital extensor sheath (CDETS); (2) to describe a single-portal endoscopic examination of the ECRS and CDETS. STUDY DESIGN: Ex vivo experimental. SAMPLE POPULATION: Thirty clinically normal cadaver thoracic equine limbs severed at the humeral diaphysis. METHODS: Ten limbs underwent plain and intrathecal contrast CT examinations and gross dissection of the ECRS and CDETS. Single-portal endoscopic examination of ECRS and CDETS was attempted in 4 limbs and endoscopic examination was performed on 16 limbs. Endoscopic video recordings were reviewed by 3 observers for quality of visualization before dissection and examination for iatrogenic damage. Interobserver agreement for ECRS and CDETS visualization was determined with Fleiss' κ agreement. RESULTS: Extensor carpi radialis sheath and CDETS anatomy was consistent between gross dissection and CT examinations. The ECRS endoscopic portal was medial at the level of the intersection between the extensor carpi obliquus and extensor carpi radialis tendon. The CDETS endoscopic portal was lateral, 5 cm proximal to the lateral styloid process of the ulna. The ECRS and CDETS were well visualized and interobserver agreement was substantial (κ = .73; P < .0001) and moderate (κ = .53; P < .0001), respectively. CONCLUSION: Computed tomography examinations provided useful anatomical information, consistent with gross dissection of the ECRS and CDETS. The described single-portal endoscopic techniques allowed consistent tenoscopic examination of the majority of the ECRS and CDETS. CLINICAL SIGNIFICANCE: Awareness of the intrathecal anatomy of the ECRS and CDETS should facilitate the treatment of these tendon sheaths. The proposed portals provide good to excellent single-site endoscopic visualization of the majority of the ECRS and CDETS.

Equine odontogenic tumors: Clinical presentation, CT findings, and outcome in 11 horses.

Odontogenic tumors present as locally invasive, slow growing, firm swellings on the face. They are rare in all species and are characterized histologically by the degree of differentiation and dental tissue of origin. Radiographic appearance is not pathognomonic for these lesions. Computed tomographic (CT) examination allows exact determination of tumor extension and aggressiveness. The objectives of this retrospective, case series study were to describe the clinical presentation, CT characteristics, and outcome in horses with histologically confirmed odontogenic tumors, and to identify imaging features suggestive of individual types of tumors. Four ameloblastomas, two ameloblastic carcinomas, three ameloblastic fibromas, and two complex odontomas were included. All but one complex odontoma presented as a single mass. All tumors were associated with maxillary or mandibular bone expansion, alveolar and cortical bone lysis, and cortical bone thinning. The majority also had cortical bone thickening and periosteal proliferation. All tumors contained some degree of mineral attenuation, although only the complex odontomas contained enamel attenuation allowing differentiation from other types of odontogenic tumors in this study. Ameloblastomas were found to have variable CT characteristics likely due to the sub-groups of ameloblastomas. Both ameloblastic carcinomas contained a mixture of mineralized and soft tissue attenuating material whereas ameloblastic fibromas were mainly composed of soft tissue attenuating material. Computed tomographic characteristics of odontogenic tumors generally indicate that they are expansile, aggressive tumors and can occur in a wide range of ages. Further investigation is needed to elucidate differences between each type of equine odontogenic tumor.

Genetic crosses and complementation reveal essential functions for the Plasmodium stage-specific actin2 in sporogonic development.

Malaria parasites have two actin isoforms, ubiquitous actin1 and specialized actin2. Actin2 is essential for late male gametogenesis, prior to egress from the host erythrocyte. Here, we examined whether the two actins fulfil overlapping functions in Plasmodium berghei. Replacement of actin2 with actin1 resulted in partial complementation of the defects in male gametogenesis and, thus, viable ookinetes were formed, able to invade the midgut epithelium and develop into oocysts. However, these remained small and their DNA was undetectable at day 8 after infection. As a consequence sporogony did not occur, resulting in a complete block of parasite transmission. Furthermore, we show that expression of actin2 is tightly controlled in female stages. The actin2 transcript is translationally repressed in female gametocytes, but translated in female gametes. The protein persists until mature ookinetes; this expression is strictly dependent on the maternally derived expression. Genetic crosses revealed that actin2 functions at an early stage of ookinete formation and that parasites lacking actin2 are unable to undergo sporogony in the mosquito midgut. Our results provide insights into the specialized role of actin2 in Plasmodium development in the mosquito and suggest that the two actin isoforms have distinct biological functions.

A perforin-like protein mediates disruption of the erythrocyte membrane during egress of Plasmodium berghei male gametocytes.

Successful gametogenesis of the malaria parasite depends on egress of the gametocytes from the erythrocytes within which they developed. Egress entails rupture of both the parasitophorous vacuole membrane and the erythrocyte plasma membrane, and precedes the formation of the motile flagellated male gametes in a process called exflagellation. We show here that egress of the male gametocyte depends on the function of a perforin-like protein, PPLP2. A mutant of Plasmodium berghei lacking PPLP2 displayed abnormal exflagellation; instead of each male gametocyte forming eight flagellated gametes, it produced gametocytes with only one, shared thicker flagellum. Using immunofluorescence and transmission electron microscopy analysis, and phenotype rescue with saponin or a pore-forming toxin, we conclude that rupture of the erythrocyte membrane is blocked in the mutant. The parasitophorous vacuole membrane, on the other hand, is ruptured normally. Some mutant parasites are still able to develop in the mosquito, possibly because the vigorous motility of the flagellated gametes eventually leads to escape from the persisting erythrocyte membrane. This is the first example of a perforin-like protein in Plasmodium parasites having a role in egress from the host cell and the first parasite protein shown to be specifically required for erythrocyte membrane disruption during egress.

Comparison of ultrasonographic and computed tomographic imaging of equine thoracolumbar articular process joints.

BACKGROUND: Osteoarthritis of the equine thoracolumbar articular process joints (APJs) has been linked to back pain. Changes are commonly diagnosed through nuclear scintigraphy, radiography and ultrasonography (US). OBJECTIVES: (1) To assess the agreement of APJ grades between US and computed tomography (CT) images; (2) to assess the effect of location on the agreement of APJ grades between US and CT images. It was hypothesised that: (1) Periarticular modelling and modification of the joint space would have the highest and lowest agreement between US and CT images, respectively; (2) Caudal thoracolumbar APJ grades would have higher agreement between US and CT images than mid thoracic APJs. STUDY DESIGN: Comparative diagnostic imaging study using cadaveric specimens. METHODS: Disarticulated thoracolumbar spines of six equids euthanised for reasons unrelated to back pain, underwent US and CT examination of the APJs. Images were assessed for periarticular modelling, modification of the joint space and enlargement of the APJ. Intra-observer, inter-modality and inter-observer agreement using multilevel weighted kappa statistics to evaluate the effects of location, US/CT characteristic and their interaction. RESULTS: Intra-observer agreement of US and CT grades ranged from moderate to perfect. Between T15 and L1, inter-modality agreement (CT vs. US) for periarticular modelling was moderate, and fair for enlargement and modification of the joint space. Inter-observer agreement (US vs. US) of periarticular modelling was substantial between T15 and L1, moderate between L1 and L5 and fair between T10 and T15. Wide confidence intervals (CIs) seen for most grades apart from inter-observer grades of periarticular modelling at T15-L5, introducing a degree of uncertainty into the results. MAIN LIMITATIONS: Large prevalence index influenced kappa values, small sample size. CONCLUSIONS: Good inter-observer (US vs. US) and inter-modality (CT vs. US) agreement of caudal thoracolumbar APJ periarticular modelling. This US characteristic provides a measure of bone change, therefore supporting the use of diagnostic ultrasound. Inter-modality CIs were wide, highlighting the study and imaging modality limitations.

Critical role for a stage-specific actin in male exflagellation of the malaria parasite.

Male gametogenesis occurs directly after uptake of malaria parasites by the mosquito vector and leads to the release of eight nucleated flagellar gametes. Here, we report that one of the two parasite actin isoforms, named actin II, is essential for this process. Disruption of actin II in Plasmodium berghei resulted in viable asexual blood stages, but male gametogenesis was specifically inhibited. Upon activation, male gametocyte DNA was replicated normally and axonemes assembled, but egress from the host cell was inhibited, and axoneme motility abolished. The major actin isoform, actin I, displayed dual localization to the cytoplasm and the nucleus in male gametocytes. After activation actin I was found to be restricted to the cytoplasm. In actII(-) mutant parasites, this re-localization was abolished and actin I remained in both cellular compartments. These findings reveal vital and pleiotropic functions for the actin II isoform in male gametogenesis of the malaria parasite.

Computed tomographic arthrography, gross anatomy and histology demonstrate a communication between synovial invaginations in the proximal aspect of the third interosseous muscle and the carpometacarpal joint in horses.

This descriptive anatomical study investigates the relationship between the third interosseous muscle, also known as the suspensory ligament, and the carpometacarpal joint in forelimbs of horses, with the hypothesis that there was a direct synovial communication between these structures as shown by computed tomographic arthrography, histology, and gross anatomy sections. Computed tomography of the carpus and metacarpal region was performed on two groups. Group 1 consisted of eight cadaver limbs undergoing computed tomographic arthrography following injection of a mixture of positive contrast medium, saline, and color-pigmented fluid solution into the middle carpal joint. Group 2 consisted of eight forelimbs assessed using plain computed tomography. The images were interpreted subjectively for contrast medium distribution and objectively by comparing Hounsfield values of the proximal suspensory ligament at 0.5 cm intervals starting at the origin and extending 3 cm distal to the proximal subchondral bone plate of the third metacarpal bone. Of the 16 limbs, two were sectioned for gross anatomy and one was documented histologically. The proximal suspensory ligament was visualized with clear margins on computed tomography images. The positive contrast medium was found within the lateral and medial lobes of the suspensory ligament in all eight (100%) limbs. Hounsfield units within the suspensory ligament following contrast injection were significantly higher than in those in the plain CT group between 0.5 and 2.5 cm distal to the proximal subchondral bone plate of the third metacarpal bone (p < 0.05). The gross anatomy sections showed color pigmentation within the suspensory ligament correlating to the contrast medium distribution evident on computed tomography images. Histology confirmed a synovial lined cavity within the suspensory ligament. The demonstration of a direct synovial communication between the internal structure of the proximal suspensory ligament and the carpometacarpal joint in horses offers further explanation for commonly encountered interactions of diagnostic local anesthesia of the carpal and subcarpal regions. When performing diagnostic or therapeutic injections into the middle carpal joint, the likely effect on the proximal suspensory ligament should be considered. Furthermore, as the proximal suspensory ligament was identified clearly on CT images, further studies are needed to elucidate the utility of CT in clinical cases with suspected soft tissue pathology in the subcarpal region.

Computed Tomographic Assessment of Individual Paranasal Sinus Compartment and Nasal Conchal Bulla Involvement in 300 Cases of Equine Sinonasal Disease.

Background: Computed tomographic (CT) imaging has allowed new anatomical studies and detailed clinical imaging of the complex, overlapping equine sinonasal structures. Despite the widespread use of CT, no study has specifically identified which compartments are most commonly affected with sinus disorders. CT has also shown the presence of intercurrent, ipsilateral nasal disorders, especially infection of the nasal conchal bullae (NCB) in many cases of sinus disease, but the frequency of intercurrent NCB infections has not been reported. Objectives: To identify which sinus compartments are most commonly affected in horses with clinical sinus disorders and to record the prevalence of NCB involvement in such cases. Study Design: Retrospective examination of CT images of horses with confirmed unilateral sinus disease. Methods: The CT images of 300 horses, from three different equine hospitals with clinically confirmed sinus disease [mainly dental (53%) and primary sinusitis (25.7%)] were retrospectively examined to determine which sinus compartments and NCBs were affected. Results: The rostral, more dependent sinus compartments were most commonly involved, i.e., the rostral maxillary sinus in 284/300 (94.7% affected) and the ventral conchal sinus (87% affected). The caudal maxillary sinus (65.3%), dorsal conchal sinus (52.7%), frontal sinus (26%), ethmoidal sinus (32%) and sphenopalatine sinus (28.7%) were less commonly affected. There was infection or destruction of the ipsilateral NCBs in 56% of horses with sinus disorders, including the ventral NCBs in 42.3%, dorsal NCBs in 29% and both NCBs in 18% of cases. Main Limitations: The horses with sinonasal disease that underwent head CT imaging include more problematic cases and horses of high value, rather than the general horse population. Conclusions: The more dependant (i.e., the RMS and VCS) sinus compartments are most commonly involved in sinus disorders, with the RMS involved in nearly every case. The more dorsally located sinuses (i.e., caudal group) are less commonly involved. Many horses with sinus disease also have disorders of their nasal conchal bullae and so the term sinonasal disease seems appropriate for these disorders.

Interaction with macrophages attenuates equine fibroblast-like synoviocyte ADAMTS5 (aggrecanase-2) gene expression following inflammatory stimulation.

The joint synovium consists of a heterogeneous cell population, chiefly comprised of macrophages, and fibroblast-like synoviocytes (FLS). An inter-species co-culture model was developed to examine interactions between these cells. Equine FLS and the canine macrophage line DH82 were differentially labeled using fluorescent markers and results from direct co-culture compared with those from both indirect co-culture, and conditioned media experiments. The transcript expression of IL-1ß, IL-6, ADAMTS4, and ADAMTS5 in each cell type were determined using species-specific qPCR assays. Lipopolysaccharide stimulation of EFLS rapidly increased IL-1ß, IL-6, ADAMTS4, and ADAMTS5 mRNAs. The induction of ADAMTS5 was significantly reduced when equine FLS were cultured with DH82 cells directly or indirectly. Exposure of equine FLS to denatured conditioned media also significantly reduced ADAMTS5 induction. DH82 cells increased interleukin-1ß expression substantially following LPS stimulation. However, knockdown of interleukin-1ß in DH82 cells, or inhibition of NF-κB in equine FLS prior to co-culture did not change the inhibitory effect on equine FLS ADAMTS5 gene expression. This work indicates that macrophages can influence FLS gene expression through a soluble mediator, and modulate the expression of an enzyme critical in osteoarthritis pathology during inflammatory stimulation. © 2018 The Authors. Journal of Orthopaedic Research® Published by WileyPeriodicals, Inc. on behalf of the Orthopaedic Research Society. J Orthop Res 9999:1-8, 2018.

Use of force preferences and perceived effectiveness of actions among Crisis Intervention Team (CIT) police officers and non-CIT officers in an escalating psychiatric crisis involving a subject with schizophrenia.

BACKGROUND: Few studies have examined police officers' use of force toward individuals with schizophrenia, despite the widely disseminated Crisis Intervention Team (CIT) model of partnership between mental health and law enforcement that seeks to reduce use of force and enhance safety of officers and individuals with mental illnesses. This study tested the hypotheses that CIT-trained officers would select a lower level of force, identify nonphysical actions as more effective, and perceive physical force as less effective in an escalating psychiatric crisis, compared with non-CIT-trained officers. METHODS: Police officers (n = 135)-48 CIT trained and 87 non-CIT trained-completed a survey containing 3 scenario-based vignettes depicting an escalating situation involving a subject with psychosis. Data were analyzed using repeated-measures analyses of variance. RESULTS: Officers escalated their preferred actions across the scenarios. A significant scenario by group interaction indicated that CIT-trained officers chose less escalation (ie, opting for less force at the third scenario) than non-CIT-trained officers. Officers reported decreasing perceived effectiveness of nonphysical action across the 3 scenarios. A significant scenario by group interaction indicated that CIT-trained officers reported a lesser decline in perceived effectiveness of nonphysical actions at the third scenario. CIT-trained officers consistently endorsed lower perceived effectiveness of physical force. CONCLUSIONS: Efforts are needed to reduce use of force toward individuals with psychotic disorders. These findings suggest that CIT may be an effective approach. In addition to clinical and programmatic implications, such findings demonstrate a role for clinicians, advocates, and schizophrenia researchers in promoting social justice through partnerships with diverse social sectors.

ATM-dependent ERK signaling via AKT in response to DNA double-strand breaks.

Ionizing radiation (IR) triggers many signaling pathways primarily originating from either damaged DNA or non-nuclear sources such as growth factor receptors. Thus, to study the DNA damage-induced signaling component alone by irradiation would be a challenge. To generate DNA double-strand breaks (DSBs) and minimize non-nuclear signaling, human cancer cells having bromodeoxyuridine (BrdU) - substituted DNA were treated with the photosensitizer Hoechst 33258 followed by long wavelength UV (UV-A) treatment (BrdU photolysis). BrdU photolysis resulted in well-controlled, dose- dependent generation of DSBs equivalent to radiation doses between 0.2 - 20 Gy, as determined by pulsed-field gel electrophoresis, and accompanied by dose-dependent ATM (ser-1981), H2AX (ser-139), Chk2 (thr-68), and p53 (ser-15) phosphorylation. Interestingly, low levels (≤ 2 Gy equivalents) of BrdU photolysis stimulated ERK phosphorylation whereas higher (> 2 Gy eq.) resulted in ERK dephosphorylation. ERK phosphorylation was ATM-dependent whereas dephosphorylation was ATM-independent. The ATM-dependent increase in ERK phosphorylation was also seen when DSBs were generated by transfection of cells with an EcoRI expression plasmid or by electroporation of EcoRI enzyme. Furthermore, AKT was critical for transmitting the DSB signal to ERK. Altogether, our results show that low levels of DSBs trigger ATM- and AKT-dependent ERK pro-survival signaling and increased cell proliferation whereas higher levels result in ERK dephosphorylation consistent with a dose-dependent switch from pro-survival to anti-survival signaling.

Pro-survival AKT and ERK signaling from EGFR and mutant EGFRvIII enhances DNA double-strand break repair in human glioma cells.

The epidermal growth factor receptor (EGFR) is frequently dysregulated in malignant glioma that leads to increased resistance to cancer therapy. Upregulation of wild type or expression of mutant EGFR is associated with tumor radioresistance and poor clinical outcome. EGFR variant III (EGFRvIII) is the most common EGFR mutation in malignant glioma. Radioresistance is thought to be, at least in part, the result of a strong cytoprotective response fueled by signaling via AKT and ERK that is heightened by radiation in the clinical dose range. Several groups including ours have shown that this response may modulate DNA repair. Herein, we show that expression of EGFRvIII promoted gamma-H2AX foci resolution, a surrogate for double-strand break (DSB) repair, and thus enhanced DNA repair. Conversely, small molecule inhibitors targeting EGFR, MEK, and the expression of dominant-negative EGFR (EGFR-CD533) significantly reduced the resolution of gamma-H2AX foci. When homologous recombination repair (HRR) and non-homologous end joining (NHEJ) were specifically examined, we found that EGFRvIII stimulated and CD533 compromised HRR and NHEJ, respectively. Furthermore, NHEJ was blocked by inhibitors of AKT and ERK signaling pathways. Moreover, expression of EGFRvIII and CD533 increased and reduced, respectively, the formation of phospho-DNA-PKcs and -ATM repair foci, and RAD51 foci and expression levels, indicating that DSB repair is regulated at multiple levels. Altogether, signaling from EGFR and EGFRvIII promotes both HRR and NHEJ that is likely a contributing factor towards the radioresistance of malignant gliomas.