Surface flow of freshwater on Adriatic islands is rare due to the extreme permeability of the karst terrain. Hence, most helminthological studies of freshwater fishes in the Adriatic drainage have focused on mainland freshwater systems, while data from islands are scarce. We collected minnow, Phoxinus lumaireul (Schinz, 1840), specimens in the Suha Ričina stream on Krk Island and screened them for helminth ectoparasites. Identification of metacercariae cysts (black spots) was carried out by sequencing part of the 28S rDNA gene, and a single monogenean worm was identified by sequencing part of the internal transcribed spacer 2 (ITS2). To estimate the level of infection, the number of black spots was counted to calculate prevalence, mean abundance, and mean intensity. Phylogenetic inference based on maximum likelihood and Bayesian inference revealed a rare black spot disease taxon, genus Uvulifer, which represents one of the first records in Europe on fish (second intermediate host), a first record from a Phoxinus host, a first record for the Adriatic drainage and Croatia, and a first record for Southern Europe in general. Furthermore, the monogenean was identified as Gyrodactylus sp., potentially representing a new species.

Background: Antibody panels are one diagnostic tool within the comprehensive evaluation of suspected autoimmune encephalitis (AIE). Over-reliance on antibody panels contributes to misdiagnosis and inflated healthcare costs. Methods: Inpatients or outpatients who had AIE antibody testing ordered from one of four adult hospitals in Calgary between January 2018 – January 2020 were included. Medical records of 150 individuals were reviewed, including those with positive antibodies or testing sent to Mayo Clinic, plus a random sample. Results: AIE antibody panels were sent for 469 individuals during the 2-year period; 42 were positive (9.0%) of which 10 were pathogenic. Of 150 individuals included in chart review, 27 (18.0%) met criteria for possible AIE at presentation and 16 (10.8%) met criteria for definite AIE at final diagnosis. Overall, antibody testing was ordered in both serum and CSF in 36.3% (versus 69.2% meeting possible AIE criteria); MRI brain was performed in 92.7% (possible AIE 92.6%), EEG in 78.7% (possible AIE 100.0%), and lumbar puncture in 66.7% (possible AIE 96.3%). A sizable proportion did not receive malignancy screening (overall 48.7%; possible AIE 29.6%). Conclusions: Antibody panels are overemphasized in the assessment for AIE and often performed unnecessarily, while other recommended clinical tests are not consistently completed.

Background: Autoantibody testing for suspected autoimmune encephalitis (AIE) in Alberta is commonly performed by Mitogen Dx (MDx) using cell-based assays (CBAs) for cell surface antibodies and line immunoassay (LA) for intracellular antibodies without confirmatory tissue immunofluorescence/immunohistochemistry (TIFF/IHC). Duplicate testing is often sent to Mayo Clinic (MC) verify, resulting in increased costs. Methods: Antibody panel results were obtained for all patients who had testing sent to both MC and MDx from adult hospitals in Calgary between 2018 and 2020. Positive antibodies were evaluated to be pathogenic/non-pathogenic by chart review and expert consensus. Results: Thirty-four individuals had antibody panels completed at both labs. Overall agreement (positive/negative panel) was fair (κ = 0.24, p =.08), even after excluding low-titre GAD65 antibodies through MC (n=9, 26.5%). MDx reported more non-pathogenic serum results, including: anti-SOX1 (n=3), anti-NMDAR (n=2) and anti-GABA(B)R (n=1). All pathogenic antibodies (n=3) were positive in both laboratories. Conclusions: No new pathogenic antibodies were identified by sending duplicate testing to MC; however, a larger number of non-pathogenic antibodies were reported by MDx, likely due to lack of confirmatory TIFF/IHC. Antibody testing for AIE should be done in labs performing confirmatory TIFF/IHC on all CBA/LA results to avoid unnecessary investigations and/or treatments.

Background: The Canadian Registry for Amyloidosis Research (CRAR) is a nationwide disease registry of transthyretin (ATTR) and light-chain (AL) amyloidosis. Recent advances in disease-modifying therapy have improved prognosis, however there is a critical need for real-world evidence to address knowledge gaps, particularly longer-term therapeutic outcomes and surveillance strategies. Methods: A multi-stakeholder process was undertaken to develop a consensus dataset for ATTR- and AL-amyloidosis. This process included surveys to rank the importance of potential data items, and a consensus meeting of the CRAR steering committee, (comprised of multidisciplinary clinical experts, and patient organization representatives). Patients and patient organizations supported the development and implementation of a patient-reported dataset. Results: Consensus data items include disease onset, progression, severity, treatments, and outcomes, as well as patient-reported outcomes. Both prospective and retrospective (including deceased) patient cohorts are included. Further baseline data will be presented on an initial cohort of patients. Conclusions: CRAR has been established to collect a longitudinal, multidisciplinary dataset that will evaluate amyloidosis care and outcomes. CRAR has launched at multiple specialty amyloidosis centers nationally and is continually expanding. The growth of this program will promote opportunities to assess real-world safety and efficacy and inform the cost-effectiveness of therapies while supporting patient recruitment for research.

Background: Several case series describe patients with refractory acetylcholine receptor antibody-positive (AChR) myasthenia gravis (MG) treated with hematopoietic stem cell transplant (HSCT). In this report, we describe four patients with anti-muscle-specific kinase (MuSK)MG treated with HSCT. Methods: We reviewed the records of all patients undergoing HSCT for MG in the Alberta Blood and Bone Marrow Transplant Program and identified 4 patients with anti-MuSK MG. Results: All 4 patients had severe disease (Myasthenia Gravis Foundation of America score IVb-V) and were refractory to multiple treatments, including rituximab. 3 patients improved with no clinical manifestations or mild symptoms and remained as such for 2, 3.5, and 5.5 years. In these 3 patients, adverse events ranged from treatable infections and transient dyspnea to persistent fatigue and premature menopause. The average worst Myasthenia Gravis Activities of Daily Living (MG-ADL) scores improved from 14.7 before to 0.3 after HSCT while their mean worst Myasthenia Gravis Quality of Life Questionnaire (MG-QoL15) scores improved from 26.7 to 0. The fourth patient developed pneumonia and passed away from respiratory failure 8 weeks post-transplant. Conclusions: In patients with severe refractory anti-MuSK MG, it may be reasonable to consider HSCT but with an appreciation of the associated risks.

OBJECTIVES/GOALS: Patients suffering from respiratory failure have few options to support oxygenation and carbon dioxide removal aside from mechanical ventilation. Our objective was to test a novel extrapulmonary mechanism of gas exchange via peritoneal oxygenated perfluorocarbon (PFC) in a large animal model. METHODS/STUDY POPULATION: Using two 50 kg swine, hypoxia was modeled with subatmospheric oxygen and hypercarbia induced with acute hypoventilation. Through a midline laparotomy, cannulas were placed into the peritoneal space to allow for PFC infusion and circulation. After abdominal closure, these cannulas were connected to a device capable of draining, oxygenating, and infusing PFC. One animal was subjected to acute hypoxia (12% FiO2) and another animal to acute hypoventilation (4 breaths per minute). Primary outcomes were times for SpO2 to reach 75 mmHg, respectively. Trials were performed without PFC and with PFC dwelling or circulating through the peritoneal space, during which abdominal and bladder pressures were monitored and maintained under 20 mmHg by regulation of the PFC volume contained in the animal. RESULTS/ANTICIPATED RESULTS: In the animal subjected to acute hypoxia (12% FiO2), survival time improved from 5:55 to 20:00 (min:sec) after 2.5 liters of oxygenated PFC was instilled in the peritoneal space. Oxygen percent saturation of PFC before and after dwelling in the peritoneal space was measured at 100% before and 70% after dwelling in the animal during this hypoxic period corresponding with a gas transfer of 300 mL of oxygen over the 20-minute trial (i.e., 15 mL/min). Continual PFC circulation did not further extend the survival time during hypoxic conditions over PFC dwelling in the abdomen. In the animal that was acutely hypoventilated, there were no detectable differences in the rate of CO2 accumulation as measured by EtCO2 or direct blood pCO2 measurements with PFC dwelling or circulating through the peritoneal space. DISCUSSION/SIGNIFICANCE: Oxygenated PFC dwelling in the peritoneal space increased the duration of systemic arterial blood saturation remaining greater than 50% during normobaric hypoxic (12% FiO2) conditions but did not appreciably clear blood carbon dioxide during hypoventilation. Future experiments will focus on maximizing the rate of systemic oxygen uptake.

A steady supply of hosts at the susceptible stage for parasitism is a major component of mass rearing parasitoids for biological control programs. Here we describe the effects of storing 5th instar Plodia interpunctella larvae in dormancy on subsequent host development in the context of host colony maintenance and effects of the duration of host dormancy on the development of Habrobracon hebetor parasitoids reared from dormant hosts. We induced dormancy with a combination of short daylength (12L:12D) and lower temperature (15°C), conditions known to induce diapause in this species, and held 5th instar larvae of P. interpunctella for a series of dormancy durations ranging from 15 to 105 days. Extended storage of dormant 5th instar larvae had no significant impacts on survival, development, or reproductive potential of P. interpunctella, reinforcing that dormant hosts have a substantial shelf life. This ability to store hosts in dormancy for more than 3 months at a time without strong negative consequences reinforces the promise of using dormancy to maintain host colonies. The proportion of hosts parasitized by H. hebetor did not vary significantly between non-dormant host larvae and dormant host larvae stored for periods as long as 105 days. Concordant with a prior study, H. hebetor adult progeny production from dormant host larvae was higher than the number of progeny produced on non-dormant host larvae. There were no differences in size, sex ratio, or reproductive output of parasitoids reared on dormant hosts compared to non-dormant hosts stored for up to 105 days. Larval development times of H. hebetor were however longer when reared on dormant hosts compared to non-dormant hosts. Our results agree with other studies showing using dormant hosts can improve parasitoid mass rearing, and we show benefits for parasitoid rearing even after 3 months of host dormancy.

Background: Transthyretin Amyloidosis (ATTR) is a common cause of both cardiomyopathy and carpal tunnel syndrome, with many patients needing carpal tunnel release (CTR). Although tafamidis is now an approved treatment of ATTR cardiomyopathy, insurers in most provinces require biopsy confirmation of amyloidosis. Endomyocardial biopsy is often the chosen approach due to optimal sensitivity, albeit with risk of serious adverse events such as stroke, cardiac tamponade, and arrhythmias. CTR may present an ideal opportunity for obtaining amyloidosis biopsy confirmation. Methods: ATTR patients undergoing CTR had biopsy of their transverse carpal ligament (TCL) and/or flexor tenosynovium to assess the sensitivity of both sites for biopsy confirmation of amyloidosis. Results: Twelve patients consecutively underwent biopsies during CTR, with 4 (33%) having bilateral CTR and biopsy. Among 16 TCL biopsies and 14 tenosynovium biopsies, 100% demonstrated amyloid deposition. Another patient had isolated tenosynovium biopsy without CTR and also demonstrated amyloidosis. There were no serious adverse events, and 1/13 (8%) had wound dehiscence requiring repeat suturing. Conclusions: Biopsy of the TCL and/or tenosynovium during CTR is safe, cost-effective, and sensitive, and may represent an alternative to endomyocardial biopsy in patients requiring tissue confirmation for tafamidis approval. ATTR patients eligible for tafamidis may benefit from early neurology assessment.

Background: Wild-type transthyretin amyloidosis (wtATTR) is an important cause of infiltrative cardiomyopathy in older adults. Carpal tunnel syndrome (CTS) is one of the most common extra-cardiac manifestations of wtATTR; however, the prevalence, severity, and risk of recurrence following carpal tunnel release (CTR) remain poorly understood. Methods: This retrospective cohort study reports findings from a single-centre experience of routine neurological screening of newly diagnosed wtATTR patients including nerve conduction studies. Consecutive wtATTR patients between 2014 and 2021 were included. Results: Seventy-nine wtATTR patients were included, 73 (92%) males, mean age of 79 years. Seventy-four (94%) had median neuropathy at the wrist (MNW), 50% having a prior diagnosis with the remaining 50% being diagnosed at screening. The majority with MNW were symptomatic (53, 67%) with moderate or severe disease (66, 84%) bilaterally (42, 53%) on electrophysiologic testing. Nineteen (24%) had recurrent CTS despite previous CTR. At the time of screening, 19 (24%) were prescribed wrist splinting and 36 (46%) were referred for CTR. Conclusions: Carpal tunnel syndrome is common in wtATTR. Most have bilateral disease with moderate to severe MNW at the time of wtATTR diagnosis. Recurrence of CTS after CTR is more common in wtATTR patients than in the general population.

Background: Light chain (AL) amyloidosis is a plasma cell disorder characterized by abnormal fibrillary light chain deposition causing cardiac, renal, hepatic, gastrointestinal and peripheral nervous system dysfunction. Muscle disease occurs in 1.5% of individuals causing progressive proximal weakness thus far considered untreatable. Methods: We reviewed two cases of AL amyloidosis associated myopathy at our institution who had robust response to plasmapheresis. Both were at stringent clinical response following CyBorME therapy during peak severity of their myopathy. Results: In case 1, a 70-year-old male with recently diagnosed kappa light chain multiple myeloma and cardiac/renal amyloidosis developed severe subacute proximal weakness preventing ambulation. CK was normal and electromyography was consistent with irritable myopathy. Deltoid biopsy showed perimysial and endomysial amyloidosis. A trial of plasmapheresis in a tapering schedule resulted in robust recovery of strength. In case 2, a 67-year-old female with recently diagnosed kappa light chain multiple myeloma with amyloidosis on fat pad aspirate developed severe subacute proximal weakness requiring prolonged hospital admission. CK was normal and electromyography demonstrated non-irritable myopathy. Bicep biopsy showed perivascular amyloidosis. A trial of plasmapheresis in a tapering schedule resulted in robust recovery of strength. Conclusions: Plasmapheresis is a novel and potentially effective treatment for patients with AL amyloidosis associated myopathy.

OBJECTIVES/GOALS: For patients suffering from respiratory failure there are limited options to support gas exchange aside from mechanical ventilation. Our goal is to design, investigate, and refine a novel device for extrapulmonary gas exchange via peritoneal perfusion with perfluorocarbons (PFC) in an animal model. METHODS/STUDY POPULATION: Hypoxic respiratory failure will be modeled using 50 kg swine mechanically ventilated with subatmospheric (10-12%) oxygen. Through a midline laparotomy, two cannulas, one for inflow and one for outflow, will be placed into the peritoneal space. After abdominal closure, the cannulas will be connected to a device capable of draining, oxygenating, regulating temperature, filtering, and pumping perfluorodecalin at a rate of 3-4 liters per minute. During induced hypoxia, the physiologic response to PFC circulation through the peritoneal space will be monitored with invasive (e.g. arterial and venous blood gases) and non-invasive measurements (e.g. pulse oximetry). RESULTS/ANTICIPATED RESULTS: We anticipate that the initiation of oxygenated perfluorocarbons perfusion through the peritoneal space during induced hypoxia will create an increase in hemoglobin oxygen saturation and partial pressure of oxygen in arterial blood. As we expect gas exchange to be occurring in the microvascular beds of the peritoneal membrane, we expect to observe an increase in the venous blood oxygen content sampled from the inferior vena cava. Using other invasive hemodynamic measures (e.g. cardiac output) and blood samples taken from multiple venous sites, a quantifiable rate of oxygen delivery will be calculable. DISCUSSION/SIGNIFICANCE: Peritoneal perfluorocarbon perfusion, if able to deliver significant amounts of oxygen, would provide a potentially lifesaving therapy for patients in respiratory failure who are unable to be supported with mechanical ventilation alone, and are not candidates for extracorporeal membrane oxygenation.

Deliberation is widely believed to enhance democracy by helping to refine the ‘public will’, moving its participants' policy attitudes closer to their ‘full-consideration’ policy attitudes – those they would hypothetically hold with unlimited information, to which they gave unlimited reflection. Yet there have also been claims that the social dynamics involved generally ‘homogenize’ attitudes (decreasing their variance), ‘polarize’ them (moving their means toward the nearer extreme), or engender ‘domination’ (moving their overall means toward those of the attitudes held by the socially advantaged) – attitude changes that may often be away from the participants' full-consideration attitudes and may thus distort rather than refine the public will. This article uses 2,601 group-issue pairs in twenty-one Deliberative Polls to examine these claims. Reassuringly, the results show no routine or strong homogenization, polarization, or domination. What little pattern there is suggests some faint homogenization, but also some faint moderation (as opposed to polarization) and opposition (as opposed to domination) – all as is to be expected when the outside-world forces shaping pre-deliberation attitudes are slightly more centrifugal than centripetal. The authors lay out a theoretical basis for these expectations and interpretations and probe the study's results, highlighting, among other things, deliberation's role in undoing outside-world effects on pre-deliberation attitudes and the observed homogenization's, polarization's, and domination's dependence on deliberative design.

Background: Newborns with hypoxic-ischemic encephalopathy (HIE) are at high risk for seizures, the majority of which have no clinical signs and therefore require continuous electroencephalographic (cEEG) monitoring for their detection. We sought to determine which neonates are at highest risk for seizures in order to optimize allocation of scarce cEEG resources. Methods: We identified term neonates diagnosed with HIE who underwent at least 24 hours of protocol-based cEEG monitoring between 2016 and 2019. We quantified seizure incidence, timing and burden, and correlated these with potential risk factors such as HIE severity, use of therapeutic hypothermia, preceding suspected clinical seizures, amplitude-integrated EEG (aEEG) background and patterns suspicious for seizures, and use of anti-seizure drugs. Results: cEEG monitoring was completed in 218 neonates with HIE, of whom 164 (75%) underwent therapeutic hypothermia. Preceding clinical/aEEG seizures occurred in 147 (67%), 99 (67%) of whom had been cooled but only 22 (10%) had cEEG-confirmed seizures. Characterization of seizure burden and correlation with potential risk factors is ongoing. Conclusions: Although seizures are commonly suspected in neonates with HIE, they are infrequently confirmed during cEEG monitoring, creating opportunities for more efficient risk-based allocation of cEEG resources.