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Supplementary Table from EGFR Inhibition Potentiates FGFR Inhibitor Therapy and Overcomes Resistance in FGFR2 Fusion–Positive Cholangiocarcinoma

Supplementary Figure from EGFR Inhibition Potentiates FGFR Inhibitor Therapy and Overcomes Resistance in FGFR2 Fusion–Positive Cholangiocarcinoma

FGFR inhibitors are approved for the treatment of advanced cholangiocarcinoma harboring FGFR2 fusions. However, the response rate is moderate, and resistance emerges rapidly due to acquired secondary FGFR2 mutations or due to other less-defined mechanisms. Here, we conducted high-throughput combination drug screens, biochemical analysis, and therapeutic studies using patient-derived models of FGFR2 fusion–positive cholangiocarcinoma to gain insight into these clinical profiles and uncover improved treatment strategies. We found that feedback activation of EGFR signaling limits FGFR inhibitor efficacy, restricting cell death induction in sensitive models and causing resistance in insensitive models lacking secondary FGFR2 mutations. Inhibition of wild-type EGFR potentiated responses to FGFR inhibitors in both contexts, durably suppressing MEK/ERK and mTOR signaling, increasing apoptosis, and causing marked tumor regressions in vivo. Our findings reveal EGFR-dependent adaptive signaling as an important mechanism limiting FGFR inhibitor efficacy and driving resistance and support clinical testing of FGFR/EGFR inhibitor therapy for FGFR2 fusion–positive cholangiocarcinoma.Significance:We demonstrate that feedback activation of EGFR signaling limits the effectiveness of FGFR inhibitor therapy and drives adaptive resistance in patient-derived models of FGFR2 fusion–positive cholangiocarcinoma. These studies support the potential of combination treatment with FGFR and EGFR inhibitors as an improved treatment for patients with FGFR2-driven cholangiocarcinoma.This article is highlighted in the In This Issue feature, p. 1171

FGFR inhibitors are approved for the treatment of advanced cholangiocarcinoma harboring FGFR2 fusions. However, the response rate is moderate, and resistance emerges rapidly due to acquired secondary FGFR2 mutations or due to other less-defined mechanisms. Here, we conducted high-throughput combination drug screens, biochemical analysis, and therapeutic studies using patient-derived models of FGFR2 fusion–positive cholangiocarcinoma to gain insight into these clinical profiles and uncover improved treatment strategies. We found that feedback activation of EGFR signaling limits FGFR inhibitor efficacy, restricting cell death induction in sensitive models and causing resistance in insensitive models lacking secondary FGFR2 mutations. Inhibition of wild-type EGFR potentiated responses to FGFR inhibitors in both contexts, durably suppressing MEK/ERK and mTOR signaling, increasing apoptosis, and causing marked tumor regressions in vivo. Our findings reveal EGFR-dependent adaptive signaling as an important mechanism limiting FGFR inhibitor efficacy and driving resistance and support clinical testing of FGFR/EGFR inhibitor therapy for FGFR2 fusion–positive cholangiocarcinoma. Significance: We demonstrate that feedback activation of EGFR signaling limits the effectiveness of FGFR inhibitor therapy and drives adaptive resistance in patient-derived models of FGFR2 fusion–positive cholangiocarcinoma. These studies support the potential of combination treatment with FGFR and EGFR inhibitors as an improved treatment for patients with FGFR2-driven cholangiocarcinoma. This article is highlighted in the In This Issue feature, p. 1171

Identifying adaptive mechanisms of metastatic cancer cells remains an elusive question in the treatment of metastatic disease, particularly in pancreatic cancer (PDA), where the majority of patients present with metastatic lesions at the time of diagnosis. A loss-of-function shRNA targeted screen in metastatic-derived cells identifiedGstt1, a member of the glutathione S-transferase superfamily, as uniquely required for metastasis and dissemination however dispensable for primary tumor growth.Gstt1is expressed in early disseminated tumor cells (DTCs), is retained within a subpopulation of slow-cycling cells within established metastases and its inhibition led to a regression of macrometastatic lesions. This distinct Gstt1highpopulation is highly metastatic and retains slow-cycling phenotypes, EMT features, and DTC characteristics compared to the Gstt1lowpopulation. Mechanistic studies indicate that in this subset of cells, Gstt1 maintains metastases by binding to and modifying intracellular fibronectin, regulating Fibronectin secretion from cancer cells and deposition into the metastatic microenvironment. We identified Gstt1 as a novel mediator of metastasis, highlighting the importance of metastatic heterogeneity and its influence on the metastatic tumor microenvironment.

Objectives: Flexor hallucis longus (FHL) tendon transfer is the method of choice in reconstructing chronic neglected Achilles tendon rupture. We performed a retrospective study to assess the incidence and degree of fatty degeneration of FHL. Material and Methods: Two hundred and twenty-five consecutive MR of ankles were reviewed retrospectively and assessed for fatty atrophy based on Goutallier classification. Results: About 42.7% had Grade 1, 8.4% had Grade 2, 3.1% had Grade 3, and 1.8% had Grade 4 fatty atrophy of FHL. Other lesions identified included posterior ankle impingement, tenosynovitis, loose bodies, and giant cell tumors of the tendon sheath. Conclusion: We suggest pre-operative radiological assessment of the FHL to establish that the FHL muscle and tendon are normal and intact and suitable for transfer surgery. We also discuss the spectrum of pathologies affecting FHL.

ObjectiveTo determine the rates of asymptomatic viral carriage and seroprevalence of SARS-CoV-2 antibodies in healthcare workers.DesignA cross-sectional study of asymptomatic healthcare workers undertaken on 24/25 April 2020.SettingUniversity Hospitals Birmingham NHS Foundation Trust (UHBFT), UK.Participants545 asymptomatic healthcare workers were recruited while at work. Participants were invited to participate via the UHBFT social media. Exclusion criteria included current symptoms consistent with COVID-19. No potential participants were excluded.InterventionParticipants volunteered a nasopharyngeal swab and a venous blood sample that were tested for SARS-CoV-2 RNA and anti-SARS-CoV-2 spike glycoprotein antibodies, respectively. Results were interpreted in the context of prior illnesses and the hospital departments in which participants worked.Main outcome measureProportion of participants demonstrating infection and positive SARS-CoV-2 serology.ResultsThe point prevalence of SARS-CoV-2 viral carriage was 2.4% (n=13/545). The overall seroprevalence of SARS-CoV-2 antibodies was 24.4% (n=126/516). Participants who reported prior symptomatic illness had higher seroprevalence (37.5% vs 17.1%, χ2=21.1034, pConclusions and relevanceWe identify differences in the occupational risk of exposure to SARS-CoV-2 between hospital departments and confirm asymptomatic seroconversion occurs in healthcare workers. Further investigation of these observations is required to inform future infection control and occupational health practices.

OBJECTIVE: Degenerative thoracic stenosis has been shown to most frequently involve the lower thoracic segments (T9-T12) where there is greater mobility and vulnerability due to flexion, extension and rotation of the spine. The thoracolumbar junction is considered anatomically to be T12-L1; the anatomical transition between the relatively immobile thoracic spine and relatively mobile lumbar spine. From anecdotal experience at our institution, we hypothesise that the true thoracolumbar junction is higher, at T10-11; the point of transition from floating to false ribs resulting in increased mobility at T10-11. METHODS AND MATERIALS: A retrospective review was performed of MRI lumbar and whole spine performed on patients aged 10–40 years in our institution over a 5-year period. Patients with previous surgery, chronic spinal disorders and congenital abnormalities were excluded from the study. Intervertebral discs from T8-9 to L1-2 were assessed for evidence of degeneration using the Pfirrmann grading system. Data obtained from a study using computer-based models to assess mean resultant loads in flexion, sitting and standing from T8-9 to L1-2 on patients aged 18–35 years was also analysed. The mean load gradients between two consecutive discs from T8 to L2 were analysed. Statistical analysis was performed with SPSS (p

Objectives: Trans-foraminal epidural injections have been used successfully to aid in the management of cervicobrachialagia. The aim of this study was to assess the cross sectional area of the vertebral artery during transforaminal cervical nerve root injections (TFCNRI) when the head is typically rotated and to compare with the neutral position cross sectional area. We hypothesize that head rotation does not lead to a change of vertebral artery calibre at the neural foramen, thus this technique can be performed relatively safely during TFCNRI. Material and Methods: A pilot study involving a retrospective review of 16 computed tomography guided TFNRI was performed and cross sectional area of vertebral artery with head tilted and head in neutral position was performed. Results: There was no correlation between the degree of head rotation and change in the area of the ipsilateral or contralateral vertebral artery. Conclusion: We suggest that head can safely be rotated to varied degrees while performing TFCNRI.

ObjectivesImproved bovine tuberculosis (bTB) diagnostics with higher sensitivity and specificity are urgently required. A better understanding of the peripheral blood transcriptional response ofMycobacterium bovis-infected animals after bovine purified protein derivative (PPD-b) stimulation of whole blood—an important component of current bTB diagnostics—will provide new information for development of better diagnostics.MethodsRNA sequencing (RNA-seq) was used to study the peripheral blood transcriptome after stimulation with PPD-b across four time points (-1 wk pre-infection, and +1 wk, +2 wk, and +10 wk post-infection) from a 14-weekM. bovisinfection time course experiment with ten age-matched Holstein-Friesian cattle.ResultsIn vitro PPD-b stimulation of peripheral blood fromM. bovis-infected and non-infected cattle elicited a strong transcriptional response. Comparison of PPD-b stimulated, and unstimulated samples revealed higher expression of genes encoding cytokine receptors, transcription factors, and interferon-inducible proteins. Lower expression was seen for genes encoding proteins involved in antimicrobial activity, C-type lectin receptors, inhibition of signal transduction, and genes encoding metal ion transporters.ConclusionsA transcriptional signature associated with the peripheral blood response to PPD-b stimulation consisting of 170 genes was identified exclusively in the post-infection time points. Therefore, this represents a panel of potential biomarkers ofM. bovisinfection.

Ankle sprains are the most frequent sport related injuries with involvement of the lateral collateral ligament complex occurring in 85% of cases. Isolated anterior talofibular ligament injury is by far the commonest followed by combined anterior talofibular and calcaneofibular ligament strain. The posterior talofibular ligament is the strongest component of the lateral collateral ligament complex and is injured in severe ankle injury along with the other lateral collateral ligaments. While isolated calcaneofibular ligament strain has been reported, calcaneofibular ligament and posterior talofibular ligament strains with an intact anterior talofibular ligament are rare and reported in cadaveric studies. We present a case of radiologically diagnosed calcaneofibular ligament and posterior talofibular ligament injury and will discuss the anatomy, stress radiography, and magnetic resonance image findings and the mechanism of this particular injury.

Xanthoma arising in bone is a very rare occurrence with only a few case reports in literature. Xanthoma arising in the spine is extremely rare with only four cases previously reported. We report a case of xanthoma of the thoracic spine in a 75-year-old male with hyperlipidemia. The imaging appearances and a review of literature will also be provided.

Groin pain can be caused by a myriad of pathologies. Abnormalities of the rectus femoris are a very rare cause of groin pain; calcific tendinopathy of the direct head is particularly so, with only two case reports in the literature. We report the first case of calcific tendinopathy of the direct head of the rectus femoris that was treated effectively with ultrasound-guided percutaneous irrigation of calcific tendinopathy (USPICT). The anatomy of the rectus femoris and the technique for US-PICT of the rectus femoris are also described.

INTRODUCTION: An intra-neural ganglion cyst of the lateral dorsal cutaneous branch of the sural nerve is rare, with only a few cases reported in the literature. MATERIALS AND METHODS: We carried out a retrospective investigation of patients with an intraneural ganglion cyst of the lateral dorsal cutaneous nerve. RESULTS: We present a case series of four patients with intra-neural ganglion cysts of the lateral dorsal cutaneous nerve, the distal continuation of the sural nerve at the lateral aspect of the foot. CONCLUSION: Intra-neural ganglion cysts of the lateral dorsal cutaneous nerve are rare. They represent a relatively uncommon source of lateral ankle pain, which can easily be diagnosed with ultrasound (US) and magnetic resonance imaging and managed effectively with US-guided aspiration or surgical excision.

Improved bovine tuberculosis (bTB) diagnostics with higher sensitivity and specificity are urgently required. A better understanding of the peripheral blood transcriptional response of Mycobacterium bovis-infected animals after bovine purified protein derivative (PPD-b) stimulation of whole blood-an important component of current bTB diagnostics-will provide new information for development of better diagnostics.RNA sequencing (RNA-seq) was used to study the peripheral blood transcriptome after stimulation with PPD-b across four time points (-1 wk pre-infection, and +1 wk, +2 wk, and +10 wk post-infection) from a 14-week M. bovis infection time course experiment with ten age-matched Holstein-Friesian cattle.In vitro PPD-b stimulation of peripheral blood from M. bovis-infected and non-infected cattle elicited a strong transcriptional response. Comparison of PPD-b stimulated, and unstimulated samples revealed higher expression of genes encoding cytokine receptors, transcription factors, and interferon-inducible proteins. Lower expression was seen for genes encoding proteins involved in antimicrobial activity, C-type lectin receptors, inhibition of signal transduction, and genes encoding metal ion transporters.A transcriptional signature associated with the peripheral blood response to PPD-b stimulation consisting of 170 genes was identified exclusively in the post-infection time points. Therefore, this represents a panel of potential biomarkers of M. bovis infection.

Background Trastuzumab is a targeted therapy for human epidermal growth factor receptor 2 (HER2)-positive breast cancer. However, trastuzumab-induced cardiotoxicity (TIC) has been reported as a single agent or combined with anthracycline. Methods such as biomarkers for early TIC detection is not available. Blood-based protein biomarker prostate-specific antigen (PSA) for diagnosis, screening, prediction of response to therapy, and disease progression has revolutionized management and outcome of prostate cancer. Nevertheless, no blood biomarkers exist for breast cancer diagnosis or screening. Methods We evaluated for the first time the potentials of cardiac myosin light chain 1(cMLC-1) as a biomarker to predict TIC, screen breast cancer and monitor breast cancer progression. Plasma cMCL-1 was measured quantitatively using enzyme-linked immunosorbent assays (ELISA). Archived paired plasma samples collected before and after trastuzumab treatment from 15 HER2 + patients with or without cardiotoxicity, recently collected unpaired plasma samples from 79 breast cancer patients (40 HER2+, 39 HER2- ) and 46 healthy donors were tested for cMLC-1 levels. Results We found that elevated plasma level of cMLC-1 is associated with cardiotoxicity in 43% of trastuzumab-treated patients. In addition, we demonstrated that elevated plasma cMCL-1 is associated with breast cancer. The cutoff cMLC-1 concentration is estimated to be 44.99 ng/mL with a sensitivity of 59.49% (95%CI: 48.47%-69.63%) and specificity of 71.74% (95%CI: 57.45% -82.68%). We also found that plasma cMCL-1 is more elevated in HER2- than in HER2 + breast cancer patients. As a result, improved sensitivity of 79.49% (95%CI: 64.47%-89.22%) with the specificity of 63.04% (95%CI:48.60%-75.48%) were obtained for cMLC-1 to predict HER2- breast cancer with the cutoff at 37.17 ng/mL. Moreover, we determined that cMLC-1 level was significantly higher in patients with metastatic breast cancer than in patients with non-metastatic breast cancer. Conclusions Here we report the first exploratory human study on the potential of cMLC-1 as a blood protein biomarker for predicting TIC. Additionally, we show our findings which shed light on and filled, to some extent, the gap of knowledge of the potential of cMLC-1 as a blood protein biomarker for screening breast cancer, especially for HER2- breast cancer, and disease progression of breast cancer.

Many aspects of photoperception by plants and microorganisms are initiated by the phytochrome (Phy) family of photoreceptors that detect light through interconversion between red light- (Pr) and far-red light-absorbing (Pfr) states. Plants synthesize a small family of Phy isoforms (PhyA to PhyE) that collectively regulate photomorphogenesis and temperature perception through redundant and unique actions. While the selective roles of these isoforms have been partially attributed to their differing abundances, expression patterns, affinities for downstream partners, and turnover rates, we show here from analysis of recombinant Arabidopsis chromoproteins that the Phy isoforms also display distinct biophysical properties. Included are a hypsochromic shift in the Pr absorption for PhyC and varying rates of Pfr to Pr thermal reversion, part of which can be attributed to the core photosensory module in each. Most strikingly, PhyB combines strong temperature dependence of thermal reversion with an order-of-magnitude faster rate to likely serve as the main physiological thermosensor, whereby thermal reversion competes with photoconversion. In addition, comparisons of Pfr occupancies for PhyA and PhyB under a range of red- and white-light fluence rates imply that low-light environments are effectively sensed by PhyA, while high-light environments, such as full sun, are effectively sensed by PhyB. Parallel analyses of the Phy isoforms from potato and maize showed that the unique features within the Arabidopsis family are conserved, thus indicating that the distinct biophysical properties among plant Phy isoforms emerged early in Phy evolution, likely to enable full interrogation of their light and temperature environments.

The SARS-CoV-2 pandemic has caused over 1 million deaths globally, mostly due to acute lung injury and acute respiratory distress syndrome, or direct complications resulting in multiple-organ failures. Little is known about the host tissue immune and cellular responses associated with COVID-19 infection, symptoms, and lethality. To address this, we collected tissues from 11 organs during the clinical autopsy of 17 individuals who succumbed to COVID-19, resulting in a tissue bank of approximately 420 specimens. We generated comprehensive cellular maps capturing COVID-19 biology related to patients’ demise through single-cell and single-nucleus RNA-Seq of lung, kidney, liver and heart tissues, and further contextualized our findings through spatial RNA profiling of distinct lung regions. We developed a computational framework that incorporates removal of ambient RNA and automated cell type annotation to facilitate comparison with other healthy and diseased tissue atlases. In the lung, we uncovered significantly altered transcriptional programs within the epithelial, immune, and stromal compartments and cell intrinsic changes in multiple cell types relative to lung tissue from healthy controls. We observed evidence of: alveolar type 2 (AT2) differentiation replacing depleted alveolar type 1 (AT1) lung epithelial cells, as previously seen in fibrosis; a concomitant increase in myofibroblasts reflective of defective tissue repair; and, putative TP63+ intrapulmonary basal-like progenitor (IPBLP) cells, similar to cells identified in H1N1 influenza, that may serve as an emergency cellular reserve for severely damaged alveoli. Together, these findings suggest the activation and failure of multiple avenues for regeneration of the epithelium in these terminal lungs. SARS-CoV-2 RNA reads were enriched in lung mononuclear phagocytic cells and endothelial cells, and these cells expressed distinct host response transcriptional programs. We corroborated the compositional and transcriptional changes in lung tissue through spatial analysis of RNA profiles in situ and distinguished unique tissue host responses between regions with and without viral RNA, and in COVID-19 donor tissues relative to healthy lung. Finally, we analyzed genetic regions implicated in COVID-19 GWAS with transcriptomic data to implicate specific cell types and genes associated with disease severity. Overall, our COVID-19 cell atlas is a foundational dataset to better understand the biological impact of SARS-CoV-2 infection across the human body and empowers the identification of new therapeutic interventions and prevention strategies.

Background In January 2020 reports of unidentified severe respiratory illness were described in Wuhan, China. A rapid expansion in cases affecting most countries around the globe led to major changes in the way people live their daily lives. In the United Kingdom, the Department of Health and Social Care directed healthcare providers to establish additional resources to manage the anticipated surge in cases that could overwhelm the health services. A priority area was testing for SARS-CoV-2 RNA and its detection by qualitative RT-PCR. Design A laboratory workflow twinning research environment with clinical laboratory capabilities was implemented and validated in the University of Birmingham within 4 days of the project initiation. The diagnostic capability was centred on an IVD CE-marked RT-PCR kit and designed to provide surge capacity to the nearby Queen Elizabeth Hospital. The service was initially tasked with testing healthcare workers (HCW) using throat swabs, and subsequently the process investigated the utility of using saliva as an alternative sample type. Results Between the 8th April 2020 and the 30th April 2020, the laboratory tested a total of 1282 HCW for SARS-CoV-2 RNA in throat swabs. RNA was detected in 54 % of those who reported symptoms compatible with COVID-19, but in only 4% who were asymptomatic. Conclusion This capability was established rapidly and utilised a cold-chain free methodology, applicable to a wide range of settings, and which can provide surge capacity and support to clinical laboratories facing increasing pressure during periods of national crisis.

Phytochromes (Phys) encompass a diverse collection of bilin-containing photoreceptors that help plants and microorganisms perceive light through photointerconversion between red light (Pr) and far-red light (Pfr)-absorbing states. In addition, Pfr reverts thermally back to Pr via a highly enthalpic process that enables temperature sensation in plants and possibly other organisms. Through domain analysis of the Arabidopsis PhyB isoform assembled recombinantly, coupled with measurements of solution size, photoconversion, and thermal reversion, we identified both proximal and distal features that influence all three metrics. Included are the downstream C-terminal histidine kinase-related domain known to promote dimerization and a conserved patch just upstream of an N-terminal Period/Arnt/Sim (PAS) domain, which upon removal dramatically accelerates thermal reversion. We also discovered that the nature of the bilin strongly influences Pfr stability. Whereas incorporation of the native bilin phytochromobilin into PhyB confers robust Pfr → Pr thermal reversion, that assembled with the cyanobacterial version phycocyanobilin, often used for optogenetics, has a dramatically stabilized Pfr state. Taken together, we conclude that Pfr acquisition and stability are impacted by a collection of opposing allosteric features that inhibit or promote photoconversion and reversion of Pfr back to Pr, thus allowing Phys to dynamically measure light, temperature, and possibly time.

Introduction Gluteus medius lesions are rare and can present with lateral hip pain. We present a spectrum of pathologies involving the gluteus medius and discuss the imaging features. Material and methods A search of our oncology database at tertiary orthopaedic oncology service was performed. The imaging features of these were analysed. Results There were nine cases involving the gluteus medius, which included soft tissue sarcomas, lipomas, infection and trauma with an average age of 62 years. Conclusion Gluteus medius lesions are rare and one should be aware of these as these can present as lateral hip pain.

Radiological imaging is a routinely used diagnostic tool in the evaluation of the orthopedic patient. Imaging modalities including conventional radiography, ultrasound (US), computed tomography (CT), magnetic resonance imaging (MRI), and nuclear medicine (NM) are routinely used in the assessment of musculoskeletal abnormalities. The choice of imaging modality used should be tailored to the individual, but patient factors, the experience and expertise of the referring clinician and the radiologist, as well as the availability, safety, cost, and invasiveness of the technique should be considered. In this chapter, we review the basic physics of each imaging modality. The role of each modality as well as its advantages and limitations in the orthopedic setting will also be highlighted.

Symmetrospora coprosmae is a red yeast from the subphylum Pucciniomycotina in the phylum Basidiomycota. Here, we present the first genome sequence of S. coprosmae strain UCD350, from an isolate collected from soil in Ireland. The genome size is 20.2 Mb.

AIM To provide a diagnostic approach to pulmonary nodules in patients with chondrosarcoma. MATERIALS AND METHODS A search of the oncology database at a specialist orthopaedic oncology referral centre was performed to identify all patients who were treated surgically for chondrosarcoma between January 2007 and December 2018. Reports from the computed tomography (CT) examinations of the thorax of these patients were reviewed. In patients who had pulmonary nodules/metastases identified on CT, data on the primary chondrosarcoma and pulmonary nodule characteristics were collected. RESULTS Twenty point two percent of patients had a pulmonary nodule identified on either initial or follow-up staging CT of the thorax, of which 8.1% were pulmonary metastases. Patients with grade 3 and dedifferentiated chondrosarcoma were more likely to have pulmonary metastases than patients with grade 1/2 chondrosarcoma. The time interval to developing metastases was shorter in patients with grade 2/3 and dedifferentiated chondrosarcoma versus patients with grade 1 chondrosarcoma. A low proportion of patients with grade 1 chondrosarcoma developed metastases (12.5%), all of which were identified at the time of a local recurrence. Nodules ≥10mm, nodules with lobulate margins, nodules containing irregular or subtle calcification, and nodules seen bilaterally or both centrally and peripherally were more likely to represent pulmonary metastases than benign nodules. CONCLUSION The diagnostic significance of pulmonary nodules (i.e., whether they represent pulmonary metastases or not) can be predicted by taking into account a number of factors, in particular, the histological grade of the patient's chondrosarcoma, the size and margins of the nodules, and the presence of subtle/irregular calcification.

The ring-shaped meniscus (RSM) is a rare meniscal variant in which there is an intermeniscal bridge between the anterior and posterior horns of the meniscus, thus forming a complete ring. The lateral meniscus is more commonly involved than the medial meniscus. Distinguishing an RSM from a bucket handle tear (BHT) with a displaced fragment in the intercondylar notch is challenging and there are reports of negative arthroscopies for suspected BHTs. There are many reports of symptomatic and asymptomatic lateral RSMs diagnosed on MRI in the literature. We report a series of RSMs in six patients, which includes MRI descriptions of two cases of medial RSM and a case of bilateral lateral RSM, which were diagnosed on MRI and have never previously been reported. This study will also describe the various imaging features of a RSM that distinguish it from a displaced BHT. We will also discuss associated rare malformations encountered in our study, such as hypoplastic anterior cruciate ligament and the anomalous insertion of the popliteus tendon onto the posterior horn of a lateral RSM.

The phytochrome (phy) family of bilin-containing photoreceptors are major regulators of plant photomorphogenesis through their unique ability to photointerconvert between a biologically inactive red light-absorbing Pr state and an active far-red light­absorbing Pfr state. While the initial steps in Pfr signaling are unclear, an early event for the phyB isoform after photoconversion is its redistribution from the cytoplasm into subnuclear foci named photobodies (PBs) that dissipate after Pfr reverts back to Pr by far-red irradiation or by temperature-dependent non-photochemical reversion. Here we present evidence that PHOTOPERIODIC CONTROL OF HYPOCOTYL 1 (PCH1) functions both as an essential structural component of phyB-containing PBs and as a direct regulator of thermal reversion that is sufficient to stabilize phyB as Pfrin vitro.By examining the genetic interaction between a constitutively active phyBY276H-YFP allele(YHB-YFP)and PCH1, we show that the loss of PCH1 prevents YHB from coalescing into PBs without affecting its nuclear localization, whereas overexpression of PCH1 dramatically increases PB levels. Loss of PCH1, presumably by impacting phyB-PB assembly, compromises a number of events elicited inYHB-YFPplants, including their constitutive photomorphogenic phenotype, red light-regulated thermomorphogenesis, and input of phyB into the circadian clock. Conversely, elevated levels of both phyB and PCH1 generate stable, yet far red-light reversible PBs that persisted for days. Collectively, our data demonstrate that the assembly of PCHl-containing PBs is critical for phyB signaling to multiple outputs, and suggest that altering PB dynamics could be exploited to modulate plant responses to light and temperature.SignificanceInArabidopsis, phytochrome B (phyB) perceives light and temperature signals to regulate various fundamental morphogenic processes in plants through its interconversion between its active Pfr and inactive Pr states. Upon photoconversion from Pr to Pfr, phyB forms subnuclear foci called photobodies, whose composition and molecular function(s) are unclear. We show here that the phyB-interacting protein PCH1 is a structural component of phyB-photobodies and protects Pfr from thermal reversion back to Pr thus helping maintain phyB signaling. Loss of PCH1 compromises photobody formation, which disrupts a number of downstream events including photo- and thermal perception and signaling into the circadian clock. These results demonstrate that forming PCHl-dependent phyB-photobodies is an essential step connecting light and temperature to controls on plant morphogenesis.

This study examines students’ interest in physics within the culture and context of the Irish education system, at both lower and upper second level. In Ireland, the majority of students (aged 12-15 years) complete an integrated science curriculum at lower second level that encompasses learning outcomes across the disciplines of physics, chemistry, biology and earth and space. Students can choose what subjects they continue with at senior cycle and typically, 13-14% of students (aged 15-18 years) continue to study physics. This study reports on the student’s interest in physics and presents collated responses from 563 lower second level science students and 142 upper second level physics students. The findings of this study denote the differences between males and females interests in studying Physics at each level and in particular, highlight the effect that school type has on females’ interest in physics.

Aim To provide a diagnostic approach to distinguish osteochondroma-induced pseudoaneurysm from sarcoma on imaging and prevent biopsy which may be fatal. Materials and Methods A retrospective study of the orthopaedic oncology database was performed to identify all patients referred to specialist orthopaedic oncology service with a final diagnosis of osteochondroma-induced pseudoaneurysm. The demographics of the cohort, the anatomical location of the osteochondromas and pseudoaneurysms, and the imaging appearances were assessed. Results Seven contemporary and one historical case of osteochondroma-induced-pseudoaneurysm were identified. There were five patients with solitary osteochondroma and three cases of hereditary multiple exostosis. Five cases underwent magnetic resonance imaging (MRI) of which pulsation artefact was present in 40%. On MRI, all the present cases demonstrated a small central pseudoaneurysm surrounded by multiple eccentric layers of thrombus and haemorrhage, which was termed the “eccentric-whorl sign”. Conclusion Osteochondroma-induced pseudoaneurysms are often misdiagnosed as malignant transformation of an osteochondroma or a soft-tissue sarcoma and referred for urgent biopsy. This study presents the largest case series, which includes the first documented case diagnosed in 1934. Differentiating and characterising pseudoaneurysms from sarcomas is imperative in order to prevent a biopsy, which could be fatal.

We describe a novel and safe needle-holding method that we have termed the ‘dual steristrip technique’. This technique can be used to stabilize the bone biopsy needle without the need for the radiologist’s hand to be in close proximity to the X-ray beam during CT-guided intervention. The dual steristrip technique uses steristrips to stabilize the bone biopsy needle and allows for accurate assessment of needle position and trajectory. This involves affixing one end of a steristrip to the skin 2 cm from the needle skin entry point, wrapping the mid-section of the steristrip around the biopsy needle and affixing the other end of the steristrip to the skin at the opposite side of the needle 2 cm from the needle skin entry point. A second steristrip is then applied in a similar fashion at 90° to the first steristrip. In our institution, we have used the dual steristrip technique to stabilize the biopsy needle in certain cases where assessment of needle position/trajectory can be more challenging. This includes cases where there is a paucity of soft tissues overlying the bone or if the bone lesion is located in the superficial cortex. We have found it to be successful in 80% of cases. The dual steristrip technique is a safe and effective needle stabilization method that should be considered by the interventional radiologist in challenging CT-guided bone biopsy cases.

Antecubital pain can be caused by a myriad of pathologies. Abnormalities of the brachialis, in particular, tendinopathy at its insertion, are extremely rare, with no case report in the literature. We report the first case of tendinopathy of the brachialis tendon at its insertion in a 42-year-old male who presented with a year-long history of antecubital pain. The anatomy of the brachialis and the technique for ultrasound-guided steroid injection of the brachialis will also be described.

Radiological imaging is an essential component of the routine evaluation of patients with musculoskeletal disorders. A variety of imaging modalities can be utilized to ensure that a correct diagnosis is reached, and the patient is put on an appropriate treatment path. In recent years, significant technological advances have been made in imaging and specific applications developed for assessing the orthopedic patient. These recent advances show promise in increasing our molecular, functional, and structural understanding of pathological processes in the musculoskeletal setting.

microRNAs (miRNAs) are a class of small non-coding endogenous RNA molecules that regulate a wide range of biological processes by post-transcriptionally regulating gene expression. Thousands of these molecules have been discovered to date, and multiple miRNAs have been shown to coordinately fine-tune cellular processes key to organismal development, homeostasis, neurobiology, immunobiology, and control of infection. The fundamental regulatory role of miRNAs in a variety of biological processes suggests that differential expression of these transcripts may be exploited as a novel source of molecular biomarkers for many different disease pathologies or abnormalities. This has been emphasized by the recent discovery of remarkably stable miRNAs in mammalian biofluids, which may originate from intracellular processes elsewhere in the body. The potential of circulating miRNAs as biomarkers of disease has mainly been demonstrated for various types of cancer. More recently, however, attention has focused on the use of circulating miRNAs as diagnostic/prognostic biomarkers of infectious disease; for example, human tuberculosis caused by infection with Mycobacterium tuberculosis, sepsis caused by multiple infectious agents, and viral hepatitis. Here, we review these developments and discuss prospects and challenges for translating circulating miRNA into novel diagnostics for infectious disease.

Although numerous models are used to evaluate the immunotoxic effects of xenobiotics on cell-mediated immunity (CMI), no holistic model for evaluating such effects on the delayed-type hypersensitivity (DTH) response has gained widespread acceptance. Due to a lack of interference from antigen-specific antibody production, the Candida albicans DTH model has recently been demonstrated to be a more appropriate model for assessing effects on CMI than other DTH models that utilize different sensitizing antigens, such as sheep erythrocytes (SRBC) or keyhole limpet hemocyanin (KLH). The present studies were conducted to validate the C. albicans DTH model for its ability to detect suppression (or the lack thereof) of CMI following exposure for 28 days to well-characterized immunosuppressive drugs, each having a different mechanism of action. The compounds evaluated included azathioprine (AZA), cyclophosphamide (CPS), cyclosporin A (CSA), dexamethasone (DEX), and the non-immunotoxic compound, benzo[e]pyrene (B[e]P)...

Mycobacterium bovis, the agent of bovine tuberculosis, causes an estimated $3 billion annual losses to global agriculture due, in part, to the limitations of current diagnostics. Development of next-generation diagnostics requires a greater understanding of the interaction between the pathogen and the bovine host. Therefore, to explore the early response of the alveolar macrophage to infection, we report the first application of RNA-sequencing to define, in exquisite detail, the transcriptomes of M. bovis-infected and non-infected alveolar macrophages from ten calves at 2, 6, 24 and 48 hours post-infection. Differentially expressed sense genes were detected at these time points that revealed enrichment of innate immune signalling functions and transcriptional suppression of host defence mechanisms (e.g., lysosome maturation). We also detected differentially expressed natural antisense transcripts, which may play a role in subverting innate immune mechanisms following infection. Furthermore, we report differential expression of novel bovine genes, some of which have immune-related functions based on orthology with human proteins. This is the first in-depth transcriptomics investigation of the alveolar macrophage response to the early stages of M. bovis infection and reveals complex patterns of gene expression and regulation that underlie the immunomodulatory mechanisms used by M. bovis to evade host defence mechanisms.

Despite sharing >99.9% genome sequence similarity at the nucleotide level, Mycobacterium tuberculosis and Mycobacterium bovis-the causative agents of human and bovine tuberculosis, respectively-exhibit distinct host preferences. M. bovis can cause disease in both cattle and humans yet rarely transmits between immuno-competent human hosts, while M. tuberculosis is a highly successful pathogen of humans that does not sustain in animal populations. Based on the key role played by alveolar macrophages during mycobacterial infection, we hypothesised that the immunological and pathological differences observed in cattle infected with virulent M. bovis and M. tuberculosis may have a basis in innate immune mechanisms; these differences, in turn, would be reflected at the macrophage mRNA and protein level. To investigate this, we have analysed the transcriptional profile of innate immune genes in bovine alveolar macrophages following 24 and 48 h infection with the genome-sequenced strains, M. bovis AF2122/97 and M. tuberculosis H37Rv. A bespoke multiplex ELISA was also used to quantify corresponding cytokine secretion in supernatants from the same infected alveolar macrophages. All cytokines showed similar significant patterns of expression (i.e., up- or down-regulation) at both the mRNA and protein levels in infected macrophages relative to parallel non-infected controls at the two time points (P ≤ 0.05). However, significant upregulation and downregulation of several innate immune genes-including TLR2, FOS, PIK3IP1, CCL4, IL1B, IL6 and TNF-and the CCL-4 protein was observed in the M. bovis-infected macrophages relative to the M. tuberculosis-infected macrophages 48 h post-infection (P ≤ 0.05). These results support the hypothesis that the divergent virulence of M. bovis and M. tuberculosis in cattle has a basis in innate immune mechanisms, which may contribute to host preference within the M. tuberculosis complex of strains.

Nanoparticle titanium dioxide (nano-TiO2) is a white pigment widely used in foods, sunscreens, and other cosmetic products. However, it remains unclear whether exposure to nano-TiO2 results in immunosuppressive effects or induces a contact hypersensitivity response. To address these data gaps, studies were conducted with the hypothesis that nano-TiO2 exposure could alter immune responses. After 28 days of oral gavage, nano-TiO2 (1.25-250 mg/kg in 0.5% methylcellulose) produced no significant effects on innate, humoral, or cell-mediated immune functions in female B6C3F1 mice. Furthermore, there were no effects on the weights of selected organs (spleen, thymus, liver, lung, and kidneys with adrenals). Following dermal exposure on the ears for 3 days, nano-TiO2 (2.5-10% w/v in 4:1 acetone:olive oil) did not affect auricular lymph node cell proliferation, although an irritancy response was observed following treatment with 5% and 10% nano-TiO2. Dermal sensitization (2.5-10%) on the back and subsequent challenge (10%) on the right ear with nano-TiO2 produced no significant effects on percentage ear swelling in the Mouse Ear Swelling Test (MEST). However, when nano-TiO2 was injected subcutaneously along the mid-line on top of the head at 125-250 mg/kg (in 0.5% methylcellulose), significant increases in auricular lymph node cell proliferation resulted. These results demonstrate that immune effects of nano-TiO2 exposure are route-of-exposure dependent, and they suggest that irritancy and/or potential hypersensitivity responses may occur following parenteral exposure or dermal administration of nano-TiO2 to compromised skin.