A risk-based approach for cell line development, manufacturing and characterization of genetically engineered, induced pluripotent stem cell-derived allogeneic cell therapies.

Advances in cellular reprogramming and gene-editing approaches have opened up the potential for a new class of ex vivo cell therapies based on genetically engineered, induced pluripotent stem cell (iPSC)-derived allogeneic cells. While these new therapies share some similarities with their primary cell-derived autologous and allogeneic cell therapy predecessors, key differences exist in the processes used for generating genetically engineered, iPSC-derived allogeneic therapies. Specifically, in iPSC-derived allogeneic therapies, donor selection and gene-editing are performed once over the lifetime of the product as opposed to as part of the manufacturing of each product batch. The introduction of a well-characterized, fully modified, clonally derived master cell bank reduces risks that have been inherent to primary-cell derived autologous and allogeneic therapies. Current regulatory guidance, which was largely developed based on the learnings gained from earlier generation therapies, leaves open questions around considerations for donor eligibility, starting materials and critical components, cell banking and genetic stability. Here, a risk-based approach is proposed to address these considerations, while regulatory guidance continues to evolve.

MRI criteria for diagnosis and predicting severity of carpal tunnel syndrome.

OBJECTIVE: To study MRI criteria for diagnosing and predicting severity of carpal tunnel syndrome (CTS). METHODS: Sixty-nine wrists in 41 symptomatic CTS patients and 32 wrists in 28 asymptomatic subjects were evaluated by MRI. Circumferential surface area (CSA), flattening ratio, relative median nerve signal intensity, and retinacular bowing were measured. CTS severity was classified as mild, moderate, or severe. Parameters for patients with and without CTS and for the three severity groups were compared. ROC curves were plotted to assess accuracy for CTS diagnosis and severity prediction. RESULTS: Significant differences were found between CTS and control wrists for median nerve CSA, flattening ratio at inlet, relative median nerve signal intensity, and retinacular bowing. ROC curve analysis revealed a sensitivity, specificity, and accuracy of median nerve CSA > 15 mm2 proximal to the tunnel (CSAp) of 85.5, 100, and 90.1%. Using either CSAp or CSAd > 15 mm2 as a diagnostic criterion, MRI could achieve a sensitivity of 100% and specificity of 94% for diagnosis of CTS while overall accuracy was 98%. Significant differences were found among the three severity groups. Sensitivity, specificity, and accuracy of prediction of severe CTS using for CSAp > 19 mm2 were 75.0, 65.9, and 69.6%, respectively. CONCLUSIONS: MRI is highly accurate at diagnosing CTS and moderately accurate at determining CTS severity. We recommend using CSA > 15 mm2 either proximal to or distal to the tunnel as a diagnostic criterion for CTS and CSA > 19 mm2 proximal to the tunnel as a marker for severe CTS.

Magnetic resonance perfusion for differentiating low-grade from high-grade gliomas at first presentation.

BACKGROUND: Gliomas are the most common primary brain tumour. They are graded using the WHO classification system, with Grade II-IV astrocytomas, oligodendrogliomas and oligoastrocytomas. Low-grade gliomas (LGGs) are WHO Grade II infiltrative brain tumours that typically appear solid and non-enhancing on magnetic resonance imaging (MRI) scans. People with LGG often have little or no neurologic deficit, so may opt for a watch-and-wait-approach over surgical resection, radiotherapy or both, as surgery can result in early neurologic disability. Occasionally, high-grade gliomas (HGGs, WHO Grade III and IV) may have the same MRI appearance as LGGs. Taking a watch-and-wait approach could be detrimental for the patient if the tumour progresses quickly. Advanced imaging techniques are increasingly used in clinical practice to predict the grade of the tumour and to aid clinical decision of when to intervene surgically. One such advanced imaging technique is magnetic resonance (MR) perfusion, which detects abnormal haemodynamic changes related to increased angiogenesis and vascular permeability, or "leakiness" that occur with aggressive tumour histology. These are reflected by changes in cerebral blood volume (CBV) expressed as rCBV (ratio of tumoural CBV to normal appearing white matter CBV) and permeability, measured by Ktrans. OBJECTIVES: To determine the diagnostic test accuracy of MR perfusion for identifying patients with primary solid and non-enhancing LGGs (WHO Grade II) at first presentation in children and adults. In performing the quantitative analysis for this review, patients with LGGs were considered disease positive while patients with HGGs were considered disease negative.To determine what clinical features and methodological features affect the accuracy of MR perfusion. SEARCH METHODS: Our search strategy used two concepts: (1) glioma and the various histologies of interest, and (2) MR perfusion. We used structured search strategies appropriate for each database searched, which included: MEDLINE (Ovid SP), Embase (Ovid SP), and Web of Science Core Collection (Science Citation Index Expanded and Conference Proceedings Citation Index). The most recent search for this review was run on 9 November 2016.We also identified 'grey literature' from online records of conference proceedings from the American College of Radiology, European Society of Radiology, American Society of Neuroradiology and European Society of Neuroradiology in the last 20 years. SELECTION CRITERIA: The titles and abstracts from the search results were screened to obtain full-text articles for inclusion or exclusion. We contacted authors to clarify or obtain missing/unpublished data.We included cross-sectional studies that performed dynamic susceptibility (DSC) or dynamic contrast-enhanced (DCE) MR perfusion or both of untreated LGGs and HGGs, and where rCBV and/or Ktrans values were reported. We selected participants with solid and non-enhancing gliomas who underwent MR perfusion within two months prior to histological confirmation. We excluded studies on participants who received radiation or chemotherapy before MR perfusion, or those without histologic confirmation. DATA COLLECTION AND ANALYSIS: Two review authors extracted information on study characteristics and data, and assessed the methodological quality using the Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2) tool. We present a summary of the study characteristics and QUADAS-2 results, and rate studies as good quality when they have low risk of bias in the domains of reference standard of tissue diagnosis and flow and timing between MR perfusion and tissue diagnosis.In the quantitative analysis, LGGs were considered disease positive, while HGGs were disease negative. The sensitivity refers to the proportion of LGGs detected by MR perfusion, and specificity as the proportion of detected HGGs. We constructed two-by-two tables with true positives and false negatives as the number of correctly and incorrectly diagnosed LGG, respectively, while true negatives and false positives are the number of correctly and incorrectly diagnosed HGG, respectively.Meta-analysis was performed on studies with two-by-two tables, with further sensitivity analysis using good quality studies. Limited data precluded regression analysis to explore heterogeneity but subgroup analysis was performed on tumour histology groups. MAIN RESULTS: Seven studies with small sample sizes (4 to 48) met our inclusion criteria. These were mostly conducted in university hospitals and mostly recruited adult patients. All studies performed DSC MR perfusion and described heterogeneous acquisition and post-processing methods. Only one study performed DCE MR perfusion, precluding quantitative analysis.Using patient-level data allowed selection of individual participants relevant to the review, with generally low risks of bias for the participant selection, reference standard and flow and timing domains. Most studies did not use a pre-specified threshold, which was considered a significant source of bias, however this did not affect quantitative analysis as we adopted a common rCBV threshold of 1.75 for the review. Concerns regarding applicability were low.From published and unpublished data, 115 participants were selected and included in the meta-analysis. Average rCBV (range) of 83 LGGs and 32 HGGs were 1.29 (0.01 to 5.10) and 1.89 (0.30 to 6.51), respectively. Using the widely accepted rCBV threshold of <1.75 to differentiate LGG from HGG, the summary sensitivity/specificity estimates were 0.83 (95% CI 0.66 to 0.93)/0.48 (95% CI 0.09 to 0.90). Sensitivity analysis using five good quality studies yielded sensitivity/specificity of 0.80 (95% CI 0.61 to 0.91)/0.67 (95% CI 0.07 to 0.98). Subgroup analysis for tumour histology showed sensitivity/specificity of 0.92 (95% CI 0.55 to 0.99)/0.42 (95% CI 0.02 to 0.95) in astrocytomas (6 studies, 55 participants) and 0.77 (95% CI 0.46 to 0.93)/0.53 (95% CI 0.14 to 0.88) in oligodendrogliomas+oligoastrocytomas (6 studies, 56 participants). Data were too sparse to investigate any differences across subgroups. AUTHORS' CONCLUSIONS: The limited available evidence precludes reliable estimation of the performance of DSC MR perfusion-derived rCBV for the identification of grade in untreated solid and non-enhancing LGG from that of HGG. Pooled data yielded a wide range of estimates for both sensitivity (range 66% to 93% for detection of LGGs) and specificity (range 9% to 90% for detection of HGGs). Other clinical and methodological features affecting accuracy of the technique could not be determined from the limited data. A larger sample size of both LGG and HGG, preferably using a standardised scanning approach and with an updated reference standard incorporating molecular profiles, is required for a definite conclusion.

Intrinsic carpal ligaments on MR and multidetector CT arthrography: comparison of axial and axial oblique planes.

PURPOSE: To compare axial and oblique axial planes on MR arthrography (MRA) and multidetector CT arthrography (CTA) to evaluate dorsal and volar parts of scapholunate (SLIL) and lunotriquetral interosseous (LTIL) ligaments. METHODS: Nine cadaveric wrists of five male subjects were studied. The visibility of dorsal and volar parts of the SLIL and LTIL was graded semi-quantitatively (good, intermediate, poor) on MRA and CTA. The presence of a ligament tear was determined on arthrosocopy and sensitivity, specificity and accuracy of tear detection were calculated. RESULTS: Oblique axial imaging was particularly useful for delineating dorsal and volar parts of the LTIL on MRA with overall 'good' visibility increased from 11 % to 78 %. The accuracy of MRA and CTA in revealing SLIL and LTIL tear was higher using the oblique axial plane. The overall accuracy for detecting SLIL tear on CTA improved from 94 % to 100 % and from 89 % to 94 % on MRA; the overall accuracy of detecting LTIL tear on CTA improved from 89 % to 100 % and from 72 % to 89 % on MRA CONCLUSION: Oblique axial imaging during CT and MR arthrography improves detection of tears in the dorsal and volar parts of both SLIL and LTIL. KEY POINTS: • Oblique axial imaging improves SLIL and LTIL visibility and tear detection. • This improvement is greater for the LTIL than for the SLIL ligament. • Overall, CT arthrography performed better than MR arthrography.

Ankle Traction During MRI of Talar Dome Osteochondral Lesions.

OBJECTIVE: The objective of our study was to assess the impact of axial traction during MRI of talar dome osteochondral lesions using a small-FOV coil. SUBJECTS AND METHODS: A prospective study of 33 patients undergoing high-resolution MRI of the ankle using a microscopy coil with and without axial traction was performed. Two radiologists independently measured the tibiotalar joint space width and semiquantitatively graded intraarticular joint fluid dispersion, cartilage surface visibility of the osteochondral lesion, and cartilage surface visibility elsewhere in the tibiotalar joint before and after traction. Patients were instructed to report any discomfort during ankle traction. RESULTS: None of the patients reported discomfort or other symptoms during ankle traction. The tibiotalar joint space significantly increased (increase in cartilage-cartilage distance, 0.5-0.7 mm; all, p < 0.05) after traction compared with before traction. The degree of intraarticular joint fluid dispersion and the cartilage surface visibility at the osteochondral lesion and elsewhere in the tibiotalar joint improved after traction (all, p < 0.05). CONCLUSION: Traction MRI of the ankle is safe and technically feasible. This study is the first to date to investigate the effect of ankle traction on the MRI assessment of talar dome osteochondral lesions. Traction improves cartilage surface visibility of talar dome osteochondral lesions.

CT pre-operative planning of a new semi-implantable bone conduction hearing device.

PURPOSE: Accommodating a novel semi-implantable bone conduction hearing device within the temporal bone presents challenges for surgical planning. This study describes the utility of CT in pre-operative assessment of such an implant. METHODS: Retrospective review of pre-operative CT, clinical and surgical records of 16 adults considered for device implantation. Radiological suitability was assessed on CT using 3D simulation software. Antero-posterior (AP) dimensions of the mastoid bone and minimum skull thickness were measured. CT planning results were correlated with operative records. RESULTS: Eight and five candidates were suitable for device placement in the transmastoid and retrosigmoid positions, respectively, and three were radiologically unsuitable. The mean AP diameter of the mastoid cavity was 14.6 mm for the transmastoid group and 4.6 mm for the retrosigmoid group (p < 0.05). Contracted mastoid and/or prior surgery were predisposing factors for unsuitability. Four transmastoid and five retrosigmoid positions required sigmoid sinus/dural depression and/or use of lifts due to insufficient bone capacity. CONCLUSION: A high proportion of patients being considered have contracted or operated mastoids, which reduces the feasibility of the transmastoid approach. This finding combined with the complex temporal bone geometry illustrates the importance of careful CT evaluation using 3D software for precise device simulation. KEY POINTS: • Preoperative temporal bone CT is essential for determining Bonebridge device suitability. • Mastoid under-pneumatisation and prior mastoidectomy predict a retrosigmoid Bonebridge position. • 3D simulation software is recommended for precise device positioning.

Radiological diagnosis and management of idiopathic spontaneous intra-abdominal haemorrhage (abdominal apoplexy): a case series.

PURPOSE: Idiopathic spontaneous intraperitoneal haemorrhage (ISIH), historically known as abdominal apoplexy, is spontaneous haemorrhage due to rupture of an intra-abdominal visceral vessel in the absence of trauma or underlying pathology. It is an exceptionally rare condition, with mostly scattered case reports available. The aim of this study was to describe this rare condition, possible associated risk factors, and usefulness of multislice-CT (MS-CT) angiogram in its diagnosis prior to intervention. METHODS: A retrospective review of patients diagnosed with ISIH. Radiological records of haemoperitoneum from a single tertiary hospital in 2006-2013 were analysed, and the cases of ISIH were identified. Demographics (including pre-morbid hypertension status), abdominal aortic calcification as a measure of atherosclerotic changes, MS-CT angiogram +/- conventional digital subtraction angiograph images, surgical records and outcomes were reviewed. RESULTS: 425 cases of haemoperitoneum were retrieved from hospital radiology database from 2006 to 2013, and 5 patients (1.1%) diagnosed with ISIH were identified (4 males, 1 female, mean age of 64 years). 4 out of 5 patients (80%) had a history of hypertension (mean 150/90 mmHg) and 3 patients had moderate abdominal aortic atherosclerosis. MS-CT angiogram was able to diagnose the bleeding source in 4 out of 5 patients, while the bleeding source remained occult in the last patient even with both MS-CT and traditional DSA angiography. Patients who underwent either embolization or surgery had no further re-bleeding in clinical follow up, ranging from 5 to 8 years. CONCLUSIONS: Hypertension and abdominal aortic atherosclerosis appear to be associated risk factors for ISIH, and MS-CT angiogram has a high sensitivity in detecting the site of haemorrhage. An integrated angiographic and surgical approach is important in managing patients with ISIH.

Harnessing Next-Generation Sequencing as a Timely and Accurate Second-Tier Screening Test for Newborn Screening of Inborn Errors of Metabolism.

In this study, we evaluated the implementation of a second-tier genetic screening test using an amplicon-based next-generation sequencing (NGS) panel in our laboratory during the period of 1 September 2021 to 31 August 2022 for the newborn screening (NBS) of six conditions for inborn errors of metabolism: citrullinemia type II (MIM #605814), systemic primary carnitine deficiency (MIM #212140), glutaric acidemia type I (MIM #231670), beta-ketothiolase deficiency (#203750), holocarboxylase synthetase deficiency (MIM #253270) and 3-hydroxy-3-methylglutaryl-CoA lyase deficiency (MIM # 246450). The custom-designed NGS panel can detect sequence variants in the relevant genes and also specifically screen for the presence of the hotspot variant IVS16ins3kb of SLC25A13 by the copy number variant calling algorithm. Genetic second-tier tests were performed for 1.8% of a total of 22,883 NBS samples. The false positive rate for these six conditions after the NGS second-tier test was only 0.017%, and two cases of citrullinemia type II would have been missed as false negatives if only biochemical first-tier testing was performed. The confirmed true positive cases were citrullinemia type II (n = 2) and systemic primary carnitine deficiency (n = 1). The false positives were later confirmed to be carrier of citrullinemia type II (n = 2), carrier of glutaric acidemia type I (n = 1) and carrier of systemic primary carnitine deficiency (n = 1). There were no false negatives reported. The incorporation of a second-tier genetic screening test by NGS greatly enhanced our program's performance with 5-working days turn-around time maintained as before. In addition, early genetic information is available at the time of recall to facilitate better clinical management and genetic counseling.

Computer-assisted patient identification tool in inborn errors of metabolism - potential for rare disease patient registry and big data analysis.

BACKGROUND: Patient registries are crucial for rare disease management. However, manual registry construction is labor-intensive and often not user-friendly. Our goal is to establish Hong Kong's first computer-assisted patient identification tool for rare diseases, starting with inborn errors of metabolism (IEM). METHODS: Patient data from 2010 to 2019 was retrieved from electronic databases. Through big data analytics, patient data were filtered based on specific IEM-related biochemical and genetic tests. Clinical notes were analyzed using a rule-based natural language processing technique called regular expression. The algorithm classified each extracted paragraph as "IEM-related" or "not IEM-related." Pathologists reviewed the paragraphs for curation, and the algorithm's performance was evaluated. RESULTS: Out of 46,419 patients with IEM-related tests, the algorithm identified 100 as "IEM-related." After pathologists' validation, 96 cases were confirmed as true IEM, with 1 uncertain case and 3 false positives. A secondary ascertainment yielded a sensitivity of 92.3% compared to our previously published IEM cohort. CONCLUSIONS: Our artificial intelligence approach provides a novel method to identify IEM patients, facilitating the creation of a centralized, computer-assisted rare disease patient registry at the local and national levels. This data can potentially be accessed by multiple stakeholders for collaborative research and to enhance healthcare management for rare diseases.

Evaluating firearm examiner conclusion variability using cartridge case reproductions.

The forensic science pattern comparison areas, including fingerprints, footwear, and firearms, have been criticized for their subjective nature. While much research has attempted to move these disciplines to more objective methods, examiners are still coming to conclusions based on their own training and experience. To complement this subjectivity, black box studies are necessary to establish the accuracy of these feature-comparison methods. However, when cartridges are fired by a firearm to create cartridge case test sets there may be significant variability within the resulting impressions. This can result in different participants receiving test sets with varying levels of difficulty based on differences in impression quality. Therefore, comparison of accuracy between examiners is not straightforward. To compare accuracy between examiners, a method called double-casting was used to create plastic cartridge case reproductions. Double-casts of twenty-one test sets of master cartridge cases were created and mailed to firearm examiners. The double-casts ensured that all participants were comparing exhibits with the same level of detail. The examiners were tasked with determining if the unknown cartridge case in each set was fired by the same firearm as the three knowns. Automated comparisons were also used to compare the cartridge cases within each set. The results from this study showed that there are differences in examiner conclusions when examining the same evidence. Furthermore, it was shown that automated comparison metrics would benefit examiners as a quality control measure to correct any potential errors and strengthen conclusions.

A comparison of ultrasound-guided rotator interval and posterior glenohumeral injection techniques for MR shoulder arthrography.

PURPOSE: The aim of this prospective, randomized study was to compare the performance of a rotator interval approach with the posterior glenohumeral approach for ultrasound-guided contrast injection prior to MR shoulder arthrography. METHOD: This study was approved by the institutional review board. One hundred and twenty consecutive patients referred for MR shoulder arthrography were randomized into four groups: rotator interval approach in-plane (n = 30); rotator interval approach out-of-plane (n = 30); posterior approach in-plane (n = 30); and posterior approach out-of plane (n = 30). Outcome measures included procedure time, number of injection attempts, patient-reported pain score (0-10), and radiologist-reported technical difficulty (0-10). MR arthrograms were assessed for adequacy of joint distension, diagnostic utility, and extra-capsular contrast leakage. RESULTS: All 120 patients had a successful ultrasound-guided injection with adequate joint distension and diagnostic utility for MR arthrography. In-plane needle guidance was less technically demanding, quicker, required fewer injection attempts, and had a lower frequency of contrast leakage than out-of-plane needle guidance. The posterior glenohumeral approach was less technically demanding though had a higher frequency of contrast leakage and caused more patient discomfort than the rotator interval approach. CONCLUSION: For ultrasound-guided shoulder joint injection, an in-plane approach is preferable. The posterior glenohumeral approach is less technically demanding though causes more patients discomfort than the rotator interval approach possibly due to the longer needle path.

Cells that produce deleterious autoreactive antibodies are vulnerable to suicide.

It is puzzling how autoreactive B cells that escape self-tolerance mechanisms manage to produce Abs that target vital cellular processes without succumbing themselves to the potentially deleterious effects of these proteins. We report that censorship indeed exists at this level: when the Ab synthesis in the cell is up-regulated in IL-6-enriched environments (e.g., adjuvant-primed mouse peritoneum), the cell dies of the increased intracellular binding between the Ab and the cellular autoantigen. In the case in which telomerase is the autoantigen, mouse hybridoma cells synthesizing such an autoantibody, which appeared to grow well in culture, could not grow in syngeneic BALB/c mice to form ascites, but grew nevertheless in athymic siblings. Culture experiments demonstrated that peritoneal cell-derived IL-6 (and accessory factors) affected the growth and functions of the hybridoma cells, including the induction of mitochondria-based apoptosis. Electron microscopy revealed an abundance of Abs in the nuclear chromatin of IL-6-stimulated cells, presumably piggy-backed there by telomerase from the cytosol. This nuclear presence was confirmed by light microscopy analysis of isolated nuclei. In two other cases, hybridoma cells synthesizing an autoantibody to GTP or osteopontin also showed similar growth inhibition in vivo. In all cases, Ab function was crucial to the demise of the cells. Thus, autoreactive cells, which synthesize autoantibodies to certain intracellular Ags, live delicately between life and death depending on the cytokine microenvironment. Paradoxically, IL-6, which is normally growth-potentiating for B cells, is proapoptotic for these cells. The findings reveal potential strategies and targets for immunotherapy.

Three-Dimensional Analysis of Cartridge Case Double-Casts.

Due to the shot-to-shot variability in tool mark reproduction on fired cartridge cases, a method of replication is needed for the creation of training and testing sets. Double-casting is one method that has been used for this application, but the accuracy and variability of this method needs to be characterized. Three firearms were used to fire 25 cartridges each to create the master cartridge cases. The double-casting method consists of creating a silicone mold of the master cartridge case. A plastic resin mix is then poured into the mold to create the double-cast reproduction. Fifteen double-casts of each of the 75 fired cartridge cases were created across different silicone molds to analyze within- and between-mold variability. The master cartridge cases and double-casts were scanned with a confocal microscope (Sensofar® S neox) to create three-dimensional representations of the surfaces. Two similarity metrics were used for the objective comparison of the double-casts to their master cartridge cases: the areal correlation coefficient (ACCFMAX ) and the number of congruent matching cells (CMC). The ACCFMAX and CMC data, along with visual examinations, showed that the double-casting method produces accurate reproductions. Within-mold variability was found to be minimal, and between-mold variability was low. These results illustrate that double-casting can be applied for training and testing purposes.

Effect of Foslip® mediated photodynamic therapy on 5-fluorouracil resistant human colorectal cancer cells.

Colorectal cancer (CRC) is the commonest cancer in Hong Kong and is often treated with 5-fluorouracil (5-FU). However the clinical application of 5-FU was limited by drug resistance in CRC. Photodynamic therapy (PDT) is a novel treatment combating CRC via the combination of photosensitizer, molecular oxygen and light activation. In this study, 5-FU resistant HT29 (HT29FU) was established and its susceptibility to Foslip® PDT tested. Effect of 5-FU to HT29 cells was measured via qPCR. Efficacy of Foslip® PDT on HT29 and HT29FU cells were measured via photosensitizer uptake, cellular localization, cytotoxicity, cell cycle distribution and signal proteins expression. 5-FU significantly induced ABCB1 mRNA expression in HT29 cells; whereas with a 24 fold increase in HT29FU cells. Both cells responded similarly to Foslip® PDT, with the inhibitory concentration IC20, IC50 and IC70 achieved at 1 ng/mL, 2 ng/mL and 5 ng/mL with 2 J/cm2 light activation respectively. Foslip® PDT triggered apoptosis and reduced JNK protein expression at IC70 on both cells. Effect of Foslip® PDT on HT29 cells was independent to 5-FU resistance properties. Therefore, Foslip® PDT could be a potential treatment for 5-FU resistant cancer patients. Further investigations on the Foslip® PDT mediated molecular changes in HT29FU cells deserve to be explored.

Percutaneous embolization of sporadic lumbar nerve root haemangioblastoma under local anaesthesia.

Pre-operative embolization of spinal tumours are mainly performed using a transarterial approach. Percutaneous embolization of spinal tumours are undertaken much less frequently, though its use has been reported in hypervascular spinal metastases1,2 and spinal paraganglioma.3 We present a patient in whom pre-operative percutaneous embolization has been performed to a recurrent lumbar nerve root haemangioblastoma that had previously been embolized using a transarterial approach. Percutaneous embolization, through targeted percutaneous puncture of the extradural component, helped reduce intraoperative blood loss, and minimize risk of spinal ischaemia.

IgM Antibodies Can Access Cryptic Antigens Denied to IgG: Hypothesis on Novel Binding Mechanism.

Antibodies are well-known protein mediators of immunity. IgM is the primordial member and the neglected sibling of the later-evolved and more proficient IgG in regard to their therapeutic and diagnostic use. Serendipitously, however, we found a paradox: While murine IgM antibodies specific for guanosine triphosphate (GTP) were able to recognize native guanylyl antigens found in primate or rat muscle tissues by immunofluorescence assays (which mimicked the auto-antibodies from autoimmune patients to skeletal or smooth muscle), the murine and human IgG counterparts failed. The results were replicated in cell-free direct binding assays using small latex microspheres decorated densely with GTP. The IgG antibodies could bind, however, if GTP was presented more spaciously on larger particles or as a univalent hapten. Accordingly, oligomerization of GTP (30-mer) destroyed the binding of the IgG antibodies but enhanced that of the IgMs in inhibition ELISA. We reason that, contrary to current belief, IgM does not bind in a lock-and-key manner like IgG. We hypothesize that whereas the intact and rigid antigen-binding site of IgG hinders the antibody from docking with antigens that are obstructed, in IgM, the two component polypeptides of the antigen-binding site can dissociate from each other and navigate individually through obstacles like the ancestral single-polypeptide antibodies found in sharks and camelids, both components eventually re-grouping around the antigen. We further speculate that polyreactive IgMs, which enigmatically bind to more than one type of antigen, use the same modus operandi. These findings call for a re-look at the clinical potential of IgM antibodies particularly in specific areas of cancer therapy, tissue pathology and vaccine design, where IgG antibodies have failed due to target inaccessibility.

Determining the number of test fires needed to represent the variability present within firearms of various calibers.

The Association of Firearm and Toolmark Examiners recommends a minimum of two test fires be performed when an unknown firearm is submitted to a laboratory prior to doing a comparison with a cartridge case collected from a crime scene. Limited research has been performed to determine how many test fires are necessary to be representative of the match distribution of a firearm. Various makes and models of firearms comprising five calibers were tested using a hybrid equivalence test to determine how many cartridge cases were required to represent the match distribution of an unknown firearm based on both breech face and firing pin correlation scores from an IBIS® HeritageTM System. The same general trend was observed for each caliber of firearm where the equivalence percentage increased from 10 to 30 cartridge cases. Overall, 15 cartridge cases are sufficient for above an 80% probability of representing the full match distribution for an unknown firearm. To approach full equivalence, 25 cartridge cases are enough because 30 cartridge cases were not found to be significantly higher in equivalence percentage for any caliber of firearm tested.

Contextual taste cues modulate olfactory learning in C. elegans by an occasion-setting mechanism.

Manipulations of context can affect learning and memory performance across species in many associative learning paradigms. Using taste cues to create distinct contexts for olfactory adaptation assays in the nematode Caenorhabditis elegans, we now show that performance in this associative learning paradigm is sensitive to context manipulations, and we investigate the mechanism(s) used for the integration of context cues in learning. One possibility is that the taste and olfactory stimuli are perceived as a combined, blended cue that the animals then associate with the unconditioned stimulus (US) in the same manner as with any other unitary conditioned stimuli (CS). Alternatively, an occasion-setting model suggests that the taste cues only define the appropriate context for olfactory memory retrieval without directly entering into the primary association. Analysis of genetic mutants demonstrated that the olfactory and context cues are sensed by distinct primary sensory neurons and that the animals' ability to use taste cues to modulate olfactory learning is independent from their ability to utilize these same taste cues for adaptation. We interpret these results as evidence for the occasion-setting mechanism in which context cues modulate primary Pavlovian association by functioning in a hierarchical manner to define the appropriate setting for memory recall.

Determining the number of test fires needed to represent the variability present within 9mm Luger firearms.

Many studies have been performed in recent years in the field of firearm examination with the goal of providing an objective method for comparisons of fired cartridge cases. No published research to support the number of test fires needed to represent the variability present within the impressions left on a cartridge case could be found. When a suspect firearm is submitted to a firearm examiner, typically two to four test fires are performed. The recovered cartridge cases are compared to each other to determine which characteristics from the firearm are reproducing, and then compared to any cartridge cases collected at a crime scene. The aim of this research was to determine the number of test fires examiners should perform when a suspect firearm is submitted to the lab to balance cartridge case acquisition time with performance accuracy. Each firearm in the IBIS® database at West Virginia University® is represented by approximately 100 fired cartridge case entries. Random samples of cartridge cases were taken separately from the breech face match score and firing pin match score lists. This subset was compared to the total match distribution of the firearm using a hybrid equivalence test to determine if the subset of similarity scores were statistically equivalent to the larger distribution of scores. For the sampled distribution to remain above 80% equivalent to the match distribution, a minimum of 15 cartridge cases should be used to model the match distribution, based on IBIS® scores. Thirty cartridge cases is a conservative estimate, allowing one to determine that the location and dispersion of the match and sampling distributions are equivalent with nearly 100% probability.

Estimation of changes in breech face and firing pin marks over consecutive discharges and its impact on an IBIS® Heritage™ System.

When a firearm is discharged, the individual marks of the breech face and firing pin are imprinted onto the primer of the cartridge case. These individual marks are reproducible between shots; however, over a large number of consecutive shots, it has been observed that minute changes in these individual marks may occur. Changes in individual marks may affect an examiner in their ability to identify or eliminate and may change the magnitude of a likelihood ratio, depending on the system used by the laboratory, but the effect that these changes have on the Integrated Ballistic Identification System (IBIS®) is largely unknown. If such changes negatively affect the performance of the IBIS® then consequences may result with respect to the correct matching candidate not be returned in the top results further compared by an examiner. Two hundred consecutive test fires performed in a clean environment (indoor shooting range) were collected in sequence from 24 new Ruger® SR9 9mm pistols and entered into an IBIS® Heritage™ System. The full known match data were extracted for each firearm, and δ sets were created that had a specific number of cartridge cases between the two cartridge cases being compared. Receiver operating characteristic (ROC) area under the curve (AUC) values for these δ sets were compared to the full known match sets to determine if any significant changes in performance resulted. Although there were instances of significant differences, these only occurred in less than 25% of comparisons, and overall no decreasing trends in performance were observed.