Epithelial-stromal crosstalk and fibrosis in eosinophilic esophagitis

Abstract

Eosinophilic esophagitis (EoE) is a food allergen-induced inflammatory disorder. EoE is increasingly recognized as a cause of swallowing dysfunction, food impaction and esophageal stricture. Inflammation of the esophageal mucosa involves immune cell infiltrate, reactive epithelial changes and fibroblast activation, culminating in robust tissue remodeling toward esophageal fibrosis characterized by excess collagen deposition in the subepithelial lamina propria. Fibrosis contributes to a unique mechanical property of the EoE-affected esophagus that is substantially stiffer than the normal esophagus. There is a great need to better understand the processes behind esophageal fibrosis in order to foster improved diagnostic tools and novel therapeutics for EoE-related esophageal fibrosis. In this review, we discuss the role of esophageal inflammatory microenvironment that promotes esophageal fibrosis, with specific emphasis upon cytokines-mediated functional epithelial-stromal interplays, recruitment and activation of a variety of effector cells, and tissue stiffness. We then explore the current state of clinical methodologies to detect and treat the EoE-related esophageal stricture.

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Tissue Acquisition in Pancreatic Cystic Lesions

Despite the progress achieved by scientific research in recent years, pancreatic cystic lesions (PCLs) remain a challenging clinical problem. A significant percentage of benign PCLs are still wrongly sent to surgery, with all the related risks of a high number of surgery-related complications and mortality. Diagnosis of the type of PCL, and risk stratification for malignancy are essential for a correct management of these lesions. Several guidelines have identified some clinical and morphological aspects suggesting the need for more accurate exams.

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Tissue Acquisition in Pancreatic Cystic Lesions

Despite the progress achieved by scientific research in recent years, pancreatic cystic lesions (PCLs) remain a challenging clinical problem. A significant percentage of benign PCLs are still wrongly sent to surgery, with all the related risks of a high number of surgery-related complications and mortality. Diagnosis of the type of PCL, and risk stratification for malignancy are essential for a correct management of these lesions. Several guidelines have identified some clinical and morphological aspects suggesting the need for more accurate exams.

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Impact of effective microorganisms on the transfer of radioactive cesium into lettuce and barley biomass

Publication date: December 2018

Source: Journal of Environmental Radioactivity, Volume 192

Author(s): Aleksander Nikolaevich Nikitin, Ihar Anatoljevich Cheshyk, Galina Zenonovna Gutseva, Elena Aleksandrovna Tankevich, Masaki Shintani, Shuichi Okumoto

Abstract

Soil microorganisms play an important role in determining the physical and chemical properties of soils. Soil microorganisms have both direct and indirect effects on the physical and chemical states of radionuclides and their availability for uptake by plant roots. Controlling the soil microorganisms to immobilize radionuclides is a promising strategy to reduce the content of radionuclides in the food chain. In this study, we evaluated the impact of effective microorganisms (EM) comprising lactic-acid bacteria, photosynthetic bacteria, and yeast on the transfer of ¹³⁷Cs into the aboveground biomass of barley and lettuce. The application of EM or fermented organic fertilizer (bokashi) alone to sod-podzolic sandy-loam soil significantly reduced the aggregated transfer factor of ¹³⁷Cs in barley by 37% and 44%, respectively. The combination of EM with bokashi or potassium fertilizer produced the largest reductions in ¹³⁷Cs transfer into barley biomass (50% and 63%, respectively). EM had a stronger effect on ¹³⁷Cs transfer into barley compared to lettuce. Laboratory experiments suggested that the effect of microorganisms on ¹³⁷Cs uptake can be attributed to a reduction in the proportion of bioavailable physicochemical forms of ¹³⁷Cs in the soils treated with EM and bokashi. This study, to the best of our knowledge, is the first to report the mechanism by which microbial fertilizers reduce the transfer of ¹³⁷Cs into plants.

Graphical abstract

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A case of interdigitating dendritic cell sarcoma studied by whole-exome sequencing

Abstract

Interdigitating dendritic cell sarcoma (IDCS) is an aggressive neoplasm and is an extremely rare disease, with a challenging diagnosis. Etiology of IDCS is also unknown and most studies with only case reports. In our case, immunohistochemistry showed that the tumor cells were positive for S100, CD45, and CD68, but negative for CD1a and CD21. This study aimed to investigate the causative factors of IDCS by sequencing the protein-coding regions of IDCS. We performed whole-exome sequencing with genomic DNA from blood and sarcoma tissue of the IDCS patient using the Illumina Hiseq 2500 platform. After that, we conducted Sanger sequencing for validation of sarcoma-specific variants and gene ontology analysis using DAVID bioinformatics resources. Through comparing sequencing data of sarcoma with normal blood, we obtained 15 nonsynonymous single nucleotide polymorphisms (SNPs) as sarcoma-specific variants. Although the 15 SNPs were not validated by Sanger sequencing due to tumor heterogeneity and low sensitivity of Sanger sequencing, we examined the function of the genes in which each SNP is located. Based on previous studies and gene ontology database, we found that POLQ encoding DNA polymerase theta enzyme and FNIP1 encoding tumor suppressor folliculin-interacting protein might have contributed to the IDCS. Our study provides potential causative genetic factors of IDCS and plays a role in advancing the understanding of IDCS pathogenesis.

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Genome-wide analyses of the Jeju, Thoroughbred, and Jeju crossbred horse populations using the high density SNP array

Abstract

The Jeju horse is an indigenous Korean horse breed that is currently registered with the Food and Agriculture Organization of the United Nations. However, there is severe lack of genomic studies on Jeju horse. This study was conducted to investigate genetic characteristics of horses including Jeju horse, Thoroughbred and Jeju crossbred (Jeju × Thoroughbred) populations. We compared the genomes of three horse populations using the Equine SNP70 Beadchip array. Short-range Linkage disequilibrium was the highest in Thoroughbred, whereas r² values were lowest in Jeju horse. Expected heterozygosity was the highest in Jeju crossbred (0.351), followed by the Thoroughbred (0.337) and Jeju horse (0.311). The level of inbreeding was slightly higher in Thoroughbred (− 0.009) than in Jeju crossbred (− 0.035) and Jeju horse (− 0.038). F_ST value was the highest between Jeju horse and Thoroughbred (0.113), whereas Jeju crossbred and Thoroughbred showed the lowest value (0.031). The genetic relationship was further assessed by principal component analysis, suggesting that Jeju crossbred is more genetically similar to Thoroughbred than Jeju horse population. Additionally, we detected potential selection signatures, for example, in loci located on LCORL/NCAPG and PROP1 genes that are known to influence body. Genome-wide analyses of the three horse populations showed that all the breeds had somewhat a low level of inbreeding within each population. In the population structure analysis, we found that Jeju crossbred was genetically closer to Thoroughbred than Jeju horse. Furthermore, we identified several signatures of selection which might be associated with traits of interest. To our current knowledge, this study is the first genomic research, analyzing genetic relationships of Jeju horse, Thoroughbred and Jeju crossbred.

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Poincaré plot indices as measures of sympathetic cardiac regulation: Responses to psychological stress and associations with pre-ejection period

Publication date: Available online 11 August 2018

Source: International Journal of Psychophysiology

Author(s): Saif Rahman, Mathew Habel, Richard J. Contrada

Abstract

The Poincaré plot is a visual representation of the R-R time series within a Cartesian plane constructed by plotting each R-R interval as a function of the previous one. The plot can be characterized by parameters that quantify the standard deviation (SD) of short- and long-term R-R interval variability (SD1 and SD2, respectively). Claims regarding the use of Poincaré plot analysis as an alternative to conventional time-domain and spectral analytic measures of parasympathetic contributions to heart rate variability (HRV) have been evaluated somewhat extensively. However, its proposed utility for characterizing sympathetic influences on HRV during periods of acute mental stress, using SD2 and the SD1/SD2 ratio, has received minimal attention. In the present study, the electrocardiogram was used to record R-R interval variability in healthy participants during a resting baseline period, while they performed a challenging reaction time task, and then during a recovery period. Simultaneously, the thoracic impedance cardiogram was used to derive the pre-ejection period (PEP), which reflects sympathetic cardiac control. HRV was quantified using time-domain, spectral analytic, and Poincaré plot indices. Task performance elicited significant reductions in HRV and PEP parameters, indicating withdrawal of parasympathetic tone with concomitant sympathetic activation. Following cessation of task performance, all HRV and impedance measures returned to baseline levels, signaling resumption of parasympathetic dominance. Bivariate associations linking SD2 and the SD1/SD2 ratio to PEP were observed; however, these were not large in magnitude, were exceeded in size by associations between the Poincaré metrics and conventional HRV markers of parasympathetic cardiac influences, and were eliminated when the latter were controlled statistically. These observations challenge the proposal that the Poincaré SD2 and SD1/SD2 metrics are suitable markers of sympathetic cardiac regulation in research concerned with mental and physical health.

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Effects of Probiotics on Inflammation and Uremic Toxins Among Patients on Dialysis: A Systematic Review and Meta-Analysis

Abstract

Background/Objectives

We performed this systematic review and meta-analysis to evaluate effects of probiotics on inflammation, uremic toxins, and gastrointestinal (GI) symptoms in end-stage renal disease (ESRD) patients.

Methods

A literature search was conducted utilizing MEDLINE, EMBASE, and Cochrane Database from inception through October 2017. We included studies that assessed assessing effects of probiotics on inflammatory markers, protein-bound uremic toxins (PBUTs), and GI symptoms in ESRD patients on dialysis. Effect estimates from the individual study were extracted and combined utilizing random effect, generic inverse variance method of DerSimonian and Laird. The protocol for this meta-analysis is registered with PROSPERO; No. CRD42017082137.

Results

Seven clinical trials with 178 ESRD patients were enrolled. There was a significant reduction in serum C-reactive protein (CRP) from baseline to post-probiotic course (≥ 2 months after treatment) with standardized mean difference (SMD) of − 0.42 (95% CI − 0.68 to − 0.16, p = 0.002). When compared to control, patients who received probiotics also had a significant higher degree of reduction in CRP level with SMDs of − 0.37 (95% CI − 0.72 to 0.03, p = 0.04). However, there were no significant changes in serum TNF-alpha or albumin with SMDs of − 0.32 (95% CI − 0.92 to 0.28, p = 0.29) and 0.16 (95% CI − 0.20 to 0.53, p = 0.39), respectively. After probiotic course, there were also significant decrease in PBUTs and improvement in overall GI symptoms (reduction in GI symptom scores) with SMDs of − 0.61 (95% CI − 1.16 to − 0.07, p = 0.03) and − 1.04 (95% CI − 1.70 to − 0.38, p = 0.002), respectively.

Conclusion

Our study demonstrates potential beneficial effects of probiotics on inflammation, uremic toxins, and GI Symptoms in ESRD patients. Future large-scale clinical studies are required to assess its benefits on other important clinical outcomes including patient mortality.

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Peak oxygen uptake, ventilatory threshold, and arterial stiffness in adolescents

Abstract

Purpose

To investigate the associations of peak oxygen uptake ( \(\dot {V}_{2{\text{peak}}}}\) ) and \(\dot {V}_2}\) at ventilatory threshold ( \(\dot {V}_2}\) at VT) with arterial stiffness in adolescents.

Methods

The participants were 55 adolescents (36 girls, 19 boys) aged 16–19 years. Aortic pulse wave velocity (PWVao) and augmentation index (AIx%) were measured by non-invasive oscillometric device from right brachial artery level. \(\dot {V}_{2{\text{peak}}}}\) was directly measured during a maximal ramp test on a cycle ergometer. \(\dot {V}_2}\) at VT was determined using the equivalents for ventilation ( \({\dot {V}_{\text{E}}}\) / \(\dot {V}{\text{C}}_2}\) and \({\dot {V}_{\text{E}}}\) / \(\dot {V}_2}\) ). \(\dot {V}_{2{\text{peak}}}}\) and \(\dot {V}_2}\) at VT were normalised for body mass (BM) and lean mass (LM). Data were analysed using linear regression analyses and analysis of covariance adjusted for age and sex.

Results

\(\dot {V}_{2{\text{peak}}}}\) normalised for BM (β = − 0.445, 95% CI − 0.783 to − 0.107) and \(\dot {V}_{2{\text{peak}}}}\) normalised for LM (β = − 0.386, 95% CI − 0.667 to − 0.106) were inversely associated with PWVao. A higher \(\dot {V}_2}\) at VT normalised for BM (β = − 0.366, 95% CI − 0.646 to − 0.087) and LM (β = − 0.321, 95% CI − 0.578 to − 0.064) was associated with lower PWVao. Adolescents in the lowest third of \(\dot {V}_{2{\text{peak}}}}\) by LM (6.6 vs. 6.1 m/s, Cohen's d = 0.33) and \(\dot {V}_2}\) at VT by LM (6.6 vs. 6.0 m/s, Cohen's d = 0.33) had a higher PWVao than those in the highest third of \(\dot {V}_{2{\text{peak}}}}\) or \(\dot {V}_2}\) at VT by LM.

Conclusions

Higher \(\dot {V}_{2{\text{peak}}}}\) and \(\dot {V}_2}\) at VT by BM and LM were related to lower arterial stiffness in adolescents. Normalising \(\dot {V}_{2{\text{peak}}}}\) and \(\dot {V}_2}\) at VT for LM would provide the most appropriate measure of cardiorespiratory fitness in relation to arterial stiffness.

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Risk of Colorectal Polyps and Malignancies Among Predominantly Rural Hispanics

Abstract

Colorectal cancer is the fourth most frequently diagnosed cancer. However, due to variations in diet, it was hypothesized that risk of adenomatous or hyperplastic polyps or malignancies would be lower among Hispanics. Participants (n = 1671) underwent a colonoscopy. Results were grouped into one of four groups: normal, hyperplastic polyps only, adenomatous polyps, and malignancies. As expected, Hispanics had a lower risk of hyperplastic (p = .031, OR = 0.47) and adenomatous polyps (p = .031, OR = 0.66) than non-Hispanic Whites. Comparison between malignancies was not possible as no Hispanics had a malignancy. Contrary to expectations, risk of hyperplastic and adenomatous polyps and malignancies were no different between non-Hispanic Blacks and Whites. Among rural and mostly rural populations, Hispanics had a lower risk of hyperplastic and adenomatous polyps.

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Genetic associations between ADHD and dopaminergic genes ( DAT1 and DRD4 ) VNTRs in Korean children

Abstract

It is well known that dopaminergic genes affect the development of attention deficit hyperactivity disorder (ADHD) in various populations. Many studies have shown that variable number tandem repeats (VNTRs) located within the 3′-untranslated region of DAT1 and in exon 3 of DRD4 are associated with ADHD development; however, these results were inconsistent. Therefore, we investigated the genetic association between two VNTRs and ADHD in Korean children. We determined the VNTRs using PCR. We examined genotype and allele frequency differences between the experimental and control groups, along with the odds ratios, using Chi square and exact tests. We observed a significant association between the children with ADHD and the control group in the 10R/10R genotype of DAT1 VNTRs (p = 0.025). In addition, the 11R allele of DAT1 VNTRs showed a higher frequency in the control group than in the ADHD group (p = 0.023). Also, the short repeat (without 11R) and long repeat alleles (including 11R) were associated with ADHD (p < 0.05). The analysis of DRD4 VNTRs revealed that the 2R allele is associated with ADHD (p = 0.025). A significant result was also observed in long and short repeats (p < 0.05). Additionally, ADHD subtypes showed that the DRD4 VNTRs are associated with combined and hyperactive-impulsive subtype groups (p < 0.05). Therefore, our results suggest that DAT1 VNTRs and DRD4 VNTRs play a role in the genetic etiology of ADHD in Korean children.

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Comparison of a Novel Cadaver Model (Fix for Life) With the Formalin-Fixed Cadaver and Manikin Model for Suitability and Realism in Airway Management Training

BACKGROUND: Manikins are widely used in airway management training; however, simulation of realism and interpatient variability remains a challenge. We investigated whether cadavers embalmed with the novel Fix for Life (F4L) embalmment method are a suitable and realistic model for teaching 3 basic airway skills: facemask ventilation, tracheal intubation, and laryngeal mask insertion compared to a manikin (SimMan 3G) and formalin-fixed cadavers. METHODS: Thirty anesthesiologists and experienced residents ("operators") were instructed to perform the 3 airway techniques in 10 F4L, 10 formalin-fixed cadavers, and 1 manikin. The order of the model type was randomized per operator. Primary outcomes were the operators' ranking of each model type as a teaching model (total rank), ranking of the model types per technique, and an operator's average verbal rating score for suitability and realism of learning the technique on the model. Secondary outcomes were the percentages of successfully performed procedures per technique and per model (success rates in completing the respective airway maneuvers). For each of the airway techniques, the Friedman analysis of variance was used to compare the 3 models on mean operator ranking and mean verbal rating scores. RESULTS: Twenty-seven of 30 operators (90%) performed all airway techniques on all of the available models, whereas 3 operators performed the majority but not all of the airway maneuvers on all models for logistical reasons. The total number of attempts for each technique was 30 on the manikin, 292 in the F4L, and 282 on the formalin-fixed cadavers. The operators' median total ranking of each model type as a teaching model was 1 for F4L, 2 for the manikin and, 3 for the formalin-fixed cadavers (P

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Continuous Popliteal-Sciatic Blocks for Postoperative Analgesia: Traditional Proximal Catheter Insertion Superficial to the Paraneural Sheath Versus a New Distal Insertion Site Deep to the Paraneural Sheath

We tested the hypothesis that during a continuous popliteal-sciatic nerve block, postoperative analgesia is improved with the catheter insertion point "deep" to the paraneural sheath immediately distal to the bifurcation between the tibial and common peroneal branches, compared with the traditional approach "superficial" to the paraneural sheath proximal to the bifurcation. The needle tip location was determined to be accurately located with a fluid bolus visualized with ultrasound; however, catheters were subsequently inserted without a similar fluid injection and visualization protocol (visualized air injection was permitted and usually implemented, but not required per protocol). The average pain (0–10 scale) the morning after surgery for subjects with a catheter inserted at the proximal subparaneural location (n = 31) was a median (interquartile) of 1.5 (0.0–3.5) vs 1.5 (0.0–4.0) for subjects with a catheter inserted at the distal supraparaneural location (n = 32; P = .927). Secondary outcomes were similarly negative. Accepted for publication July 2, 2018. Funding: This work was supported by the Department of Anesthesiology, University California San Diego (San Diego, CA). The authors declare no conflicts of interest. Institutional review board: University of California, San Diego (UCSD) Human Research Protections Program at 9500 Gilman Dr, Mail Code 0052, La Jolla, CA 92093–0052; (858) 657–5100. E-mail: tnelson@ucsd.edu. This report describes human research. This study was conducted with written informed consent from the study subjects. This report describes a prospective randomized clinical trial. The authors state that the report includes every item in the Consolidated Standards of Reporting Trials (CONSORT) checklist for a prospective randomized clinical trial. The study was registered before patient enrollment. This was not an observational clinical study. Registry URL: clinicaltrials.gov NCT02523235. Plain Language Summary: We found no evidence that during a "continuous" popliteal nerve block, postoperative analgesia is improved with the perineural catheter tip deep to the paraneural sheath immediately distal to the sciatic bifurcation, compared with a traditional approach with the catheter tip superficial to the paraneural sheath proximal to the bifurcation. The contents of this article are solely the responsibility of the authors and do not necessarily represent the official views of the funding entity. Reprints will not be available from the authors. Address correspondence to Brian M. Ilfeld, MD, MS (Clinical Investigation), Department of Anesthesiology, University California, San Diego, 200 W Arbor Dr, MC 8770, San Diego, CA, 92103. Address e-mail to bilfeld@ucsd.edu. © 2018 International Anesthesia Research Society

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Status of Women in Academic Anesthesiology: A 10-Year Update

BACKGROUND: Gender inequity is still prevalent in today's medical workforce. Previous studies have investigated the status of women in academic anesthesiology. The objective of this study is to provide a current update on the status of women in academic anesthesiology. We hypothesized that while the number of women in academic anesthesiology has increased in the past 10 years, major gender disparities continue to persist, most notably in leadership roles. METHODS: Medical student, resident, and faculty data were obtained from the Association of American Medical Colleges. The number of women in anesthesiology at the resident and faculty level, the distribution of faculty academic rank, and the number of women chairpersons were compared across the period from 2006 to 2016. The gender distribution of major anesthesiology journal editorial boards and data on anesthesiology research grant awards, among other leadership roles, were collected from websites and compared to data from 2005 and 2006. RESULTS: The number (%) of women anesthesiology residents/faculty has increased from 1570 (32%)/1783 (29%) in 2006 to 2145 (35%)/2945 (36%) in 2016 (P = .004 and P

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Diagnostic Testing and Decision-Making: Beauty Is Not Just in the Eye of the Beholder

To use a diagnostic test effectively and consistently in their practice, clinicians need to know how well the test distinguishes between those patients who have the suspected acute or chronic disease and those patients who do not. Clinicians are equally interested and usually more concerned whether, based on the results of a screening test, a given patient actually: (1) does or does not have the suspected disease; or (2) will or will not subsequently experience the adverse event or outcome. Medical tests that are performed to screen for a risk factor, diagnose a disease, or to estimate a patient's prognosis are frequently a key component of a clinical research study. Like therapeutic interventions, medical tests require proper analysis and demonstrated efficacy before being incorporated into routine clinical practice. This basic statistical tutorial, thus, discusses the fundamental concepts and techniques related to diagnostic testing and medical decision-making, including sensitivity and specificity, positive predictive value and negative predictive value, positive and negative likelihood ratio, receiver operating characteristic curve, diagnostic accuracy, choosing a best cut-point for a continuous variable biomarker, comparing methods on diagnostic accuracy, and design of a diagnostic accuracy study. This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal. Accepted for publication July 3, 2018. Funding: None. The authors declare no conflicts of interest. Reprints will not be available from the authors. Address correspondence to Thomas R. Vetter, MD, MPH, Department of Surgery and Perioperative Care, Dell Medical School at the University of Texas at Austin, Health Discovery Bldg, Room 6.812, 1701 Trinity St, Austin, TX 78712. Address e-mail to thomas.vetter@austin.utexas.edu. © 2018 International Anesthesia Research Society

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Narrative Review of Decision-Making Processes in Critical Care

Several theories describing the decision-making process in the intensive care unit (ICU) have been formulated. However, none of them appreciate the complexities of the process in an eclectic way by unifying several miscellaneous variables in 1 comprehensive theory. The purpose of this review is to highlight the key intricacies associated with the decision-making process in the ICU, to describe the theoretical frameworks with a special emphasis on gaps of knowledge, and to offer some avenues for improvement. The application of theoretical framework helps us to understand and to modify the structure of the process. Expected utility theory, regret theory, prospect theory, fuzzy-trace theory, construal level theory, and quantum probability theory were formulated over the years to appreciate an increased complexity of the decision-making process in the ICU. However the decision makers engage, these models may affect patient care because each of these frameworks has several benefits and downsides. There are gaps of knowledge in understanding how physicians match the different theoretical frameworks of the decision-making process with the potentially high ICU variability and load, especially when the "best outcome" is often nondiscrete and multidimensional. Furthermore, it is unclear when the preferential application of reflexive, habitual, variable, and biased-prone processes results in patient and staff detriment. We suggest better matching of theoretical frameworks with strengths of the human decision-making process and balanced application computer aids, artificial intelligence, and organizational modifications. The key component of this integration is work to increase the self-awareness of decision-making processes among residents, fellows, and attending physicians. Accepted for publication June 19, 2018. Funding: None. The authors declare no conflicts of interest. Reprints will not be available from the authors. Address correspondence to Krzysztof Laudanski, MD, PhD, AM FCCM, Department of Anesthesiology and Critical Care, Perelman School of Medicine at the University of Pennsylvania, 3620 Hamilton Walk, Philadelphia, PA 19104. Address e-mail to klaudanski@gmail.com. © 2018 International Anesthesia Research Society

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Using the radioligand-receptor binding assay for paralytic shellfish toxins: A case study on shellfish from Morocco

Publication date: December 2018

Source: Journal of Environmental Radioactivity, Volume 192

Author(s): J. Naouli, R. Abouabdellah, A. Bennouna, A. Laissaoui, P.W. Swarzenski, H. Ait Bouh, A. Mesfioui, M.-S. Benbrahim, M.-Y. Dechraoui Bottein

Abstract

Paralytic shellfish poisoning (PSP) events occur regularly along the Mediterranean and Atlantic coast of Morocco, and have been responsible for several severe cases of human intoxication. Along the southern Atlantic coast of Morocco, aquaculture and intensive artisanal fishing practices have recently been particularly heavily impacted, and toxic species have been observed in increasing intensity and frequency. In the 1990's a regulatory monitoring program was established for the coastal waters off Morocco by the National Institute of Fisheries Research (INRH), to reduce the risk of intoxication with biotoxins. The regulatory monitoring is conducted weekly and includes toxic phytoplankton enumeration and identification, as well as saxitoxin (STX) analysis in seafood using the mouse bioassay (MBA). Animal testing remains the most widely used screening method for PSP toxin detection, yet its use is being reconsidered for animal-related ethical issues, as well as for practical considerations. To be able to better evaluate alternatives to animal testing, the performance of a nuclear-based radioligand-receptor binding assay (RBA) for paralytic shellfish toxins was assessed and compared with the MBA using four commercially important shellfish matrices, including cockles Cerastoderma edule, razor shells Solen marginatus, oysters Crassostrea gigas, and mussels Perna perna.

Over 50 samples were collected and analysed as part of the regulatory monitoring framework including a suite of monthly samples from 2017 and all samples identified as toxic by MBA since 2011. Testing of reference material and evaluation of assay-critical parameters (e.g. slope of calibration curve, internal quality control QC and IC50) confirmed the robustness of the RBA methodology. With this RBA method, STX-like activity detected in shellfish samples ranged from 33 to 8500 μg STX equivalents per kg. RBA data were significantly correlated (P < 0.0001, Pearson r = 0.96) with the MBA-derived dataset. Importantly, the RBA method allowed for the detection and quantification of PSP toxins at levels not detectable by using the mouse bioassay. The limits of quantification of the RBA was calculated and found to be 10-fold lower than that of the MBA, respectively 35.24 ± 5.99 and 325 μg STX equivalents per kg of tissue. In addition, the RBA was easier to use and produced reliable results more rapidly than the MBA and without use of live animals.

Considering the increasing risks associated with harmful algal blooms, globally and in Morocco, together with the increased development of aquaculture production and seafood consumption and the difficulties of live animal testing, these findings indicate that the RBA method is a reliable and effective alternative to the MBA method.

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