Authentic Specimens Research Articles

LC-MS/MS confirmation of 11-nor-9-carboxy-tetrahydrocannabinol (Δ8, Δ9, Δ1°) and hexahydrocannabinol (HHC) metabolites in authentic urine specimens.

Recently, tetrahydrocannabinol (THC) isomers and other semi-synthetic cannabinoids have been introduced into the consumer market as alternatives to botanical cannabis. To assess the prevalence of these potential new analytical targets, a liquid chromatography-tandem mass spectrometry confirmation method was developed for the quantitation of seven cannabinoid metabolites and the qualitative identification of four others in urine. The validated method was applied to authentic urine specimens that screened positive by immunoassay (50 ng/mL cutoff; n=1300). The most commonly observed analytes were 11-nor-9-carboxy-Δ8- and Δ9-THC (Δ8- and Δ9-THCCOOH), with the combination of the two seen as the most prominent analyte combination found. In addition to these metabolites, Δ1°-THCCOOH was observed in 77 specimens. This is the first study to report Δ1°-THCCOOH in authentic urine specimens, with this analyte always appearing in combination with Δ9-THCCOOH. Cross-reactivity studies were performed for (6aR,9R)-Δ1°-THCCOOH using the Beckman Coulter Emit® II Plus Cannabinoid immunoassay and demonstrated cross reactivity equivalent to the Δ9-THCCOOH cutoff, providing added confidence in the reported prevalence and detection patterns. Additionally, 11-nor-9(R)-carboxy-hexahydrocannabinol (9(R)-HHCCOOH) was the most abundant stereoisomer (n=12) in specimens containing HHC metabolites alone (n=14). This is in contrast to 9(S)-HHCCOOH, which was the predominant stereoisomer in specimens containing Δ8- and/or Δ9-THCCOOH. Although HHC and Δ1°-THC metabolites are emerging toxicology findings, based on these specimens collected between April 2022 and May 2024, an analytical panel containing Δ8- and Δ9-THCCOOH appears to be sufficient for revealing cannabinoid exposure within workplace monitoring and deterrence programs.

The Collection of typical and authentic specimens of vascular plants in the Herbarium of the Federal Scientific Center for Biodive rsity, Far Eastern Branch of the Russian Academy of Sciences (VLA)

The Collection of typical and authentic specimens of vascular plants in the Herbarium of the Federal Scientific Center for Biodive rsity, Far Eastern Branch of the Russian Academy of Sciences (VLA)

B-149 Diverse glycosylation of circulating alpha fetoprotein in chronic liver diseases profiled by intact mass high resolution mass spectrometry

Abstract Background Alpha fetoprotein (AFP) is a circulating tumor marker utilized clinically to detect liver cirrhosis, liver cancer and testicular seminomas. AFP molecule contains one single N-glycosylation site, with highly variable glycan structures, resulting in diverse AFP glycoforms in circulating blood. The core fucosylation of AFP (AFP-L3) is a clinical biomarker to monitor and predict risks of liver cancer progression and recurrence. Clinical testing for AFP-L3 utilizes lectin-binding and gel-shift electrophoresis. We developed a novel method to measure AFP glycoforms, based on immune-enrichment and liquid chromatography with high resolution mass spectrometry (LC-HRMS). Methods Immune enrichment of AFP was achieved by anti-AFP antibody, immobilized to magnetic beads. After binding and enrichment, the magnetic beads were washed, AFP was eluted, and the samples were analyzed by LC-HRMS (Exploris 480 MS and Vanquish UHPLC, Thermo Fisher Scientific), using full scan, positive polarity acquisition mode. Chromatographic separation was performed on PLRP column (Agilent); flow rate 200 µL/min. The HRMS data were processed and deconvoluted with intact mass analysis workflow in Biopharma Finder 5.1 (Thermo Fisher Scientific) to derive the intact AFP masses. The method’s sensitivity and imprecision were assessed by replicate analysis of serial diluted patient serum samples. AFP and its associated glycoforms were matched and identified with an in-house developed bioinformatics workflow. The deconvoluted intact masses and the sum intensity of detected charged species for each AFP glycoform were used to quantify relative abundance of AFP glycoforms. Results The method specificity for AFP was confirmed by bottom-up analysis after immune-enrichment with observed peptide sequence coverage over 35%. The recovery of the immune enrichment was shown to be independent of the AFP glycoform compositions. The identified AFP glycoforms and their relative abundance were consistent among replicates between AFP concentration 100 ng/mL and 5000 ng/mL. Analysis of authentic serum specimens from patients with different chronic liver diseases and liver cancer (n=102), revealed multiple complex biantennary branched AFP glycan modifications, with different numbers of terminal sialic acid and core fucose. Four AFP glycoforms were identified in over 60% patient samples with elevated AFP concentrations. Although occurring consistently, relative abundance of these AFP glycoforms varied significantly among patients. Assessment of the AFP fucosylation and sialylation features suggested that the two are orthogonal processes. Clinical correlations suggested that fewer terminal sialic acids were associated with impaired liver functions, while the fucosylation was linked to inflammatory responses and presence of liver malignancy. Strong linear correlation (Spearman’s rs=0.952, N=35) was observed between AFP-L3% quantitated by the LC-HRMS method, and AFP-L3% determined by the lectin-binding based isotachophoresis assay (Wako Diagnostics). Discrepancies between the two methods were observed in specimens containing < 400 ng/mL of AFP, in which the LC-HRMS method detected only a single predominant glycoform. Conclusions We developed and evaluated the LC-HRMS method for profiling circulating AFP glycoforms in patients with chronic liver diseases and liver cancer. The detected AFP glycoforms and their relative abundance were used to establish glycan features, which can be further linked to cancer development, impaired liver function, and inflammation.

Assessment of Δ9-THC and Δ9-THCCOOH bias, precision, and ionization suppression/enhancement between solid tissue homogenate and supernatant by LC-MS-MS.

Liquid chromatography-triple quadrupole mass spectrometry (LC-MS-MS) assays are frequently utilized for screening and confirmatory purposes in the forensic toxicology laboratory. While these techniques are excellent for the targeted identification and quantitation of a wide variety of drug classes, validation and determining fit-for-purpose is a significant requirement for each method. In the USA, the American National Standards Institute and Academy Standards Board first edition of Standard 036 currently serves as a primary resource in forensic toxicology method validation and mandates that laboratories evaluate critical performance characteristics to help ensure the production of forensically defensible results. Due to the variability of specimen quality frequently encountered in the discipline of postmortem toxicology, the State of Maryland Office of the Chief Medical Examiner Forensic Toxicology Laboratory routinely analyzes solid tissue specimens as part of the medicolegal death investigation process and evaluates liver as a representative solid tissue matrix during method validation. Authentic postmortem specimens (e.g. liver, kidney, skeletal muscle, and spleen) were used to investigate the effects of analyzing solid tissue homogenate versus solid tissue supernatant on bias, precision, and ionization suppression/enhancement of Δ9-THC and Δ9-THCCOOH. Bias was <20% for Δ9-THC and Δ9-THCCOOH in liver homogenate and supernatant with a single exception of the low QC concentration for Δ9-THC in liver homogenate (-29%). Within-run and between-run CV was <20% for Δ9-THC and Δ9-THCCOOH in liver homogenate and supernatant. Δ9-THC and Δ9-THC-d3 exhibited significant ion suppression in both liver homogenate and supernatant, while Δ9-THCCOOH and Δ9-THCCOOH-d3 showed both ion suppression and enhancement in these matrices. Noticeable quantitative differences were observed in authentic postmortem solid tissue homogenate and supernatant specimens despite evaluating identical tissue samplings. A brief discussion of the results is presented using a validated LC-MS-MS method for the confirmation and quantitation of Δ9-THC and Δ9-THCCOOH in postmortem casework.

Pharmacognostical Study of Root Bark of &lt;I&gt;Erythrina indica&lt;/I&gt; Lam. Paribhadra

Paribhadra, one of the reputed drugs of the indigenous system of medicine, is equated with Erythrina indica Lamk. Syn. E. variegata Linn. var. orientalis Linn., Family Fabaceae. Some other species of Erythrina viz. E. suberosa Roxb. and E. stricta Roxb., have also been mentioned as 'Paribhadra in different text books on Indian Medicine and are used as substitutes for the genuine drug, Erythrina indica.The pharmacognostical study is made of the root and root bark of E. indica Lamk., as based on material collected from authentic specimens and covers macro- and microscopical characters highlighting the diagnostic features, micro-chemical tests, examination of the powdered drug and preliminary phytochemical investigation.

Automated extraction and LC-MS-MS analysis of 11-nor-9-carboxy-tetrahydrocannabinol isomers and prevalence in authentic urine specimens.

11-Nor-9-carboxy-Δ9-tetrahydrocannabinol (Δ9-THCCOOH) is the most frequently detected illicit drug metabolite in the military drug testing program. An increasing number of specimens containing unresolved Δ8-THCCOOH prompted the addition of this analyte to the Department of Defense drug testing panel. A method was developed and validated for the quantitative confirmation of the carboxylated metabolites of Δ8- and Δ9-THC in urine samples utilizing automated pipette tip dispersive solid-phase extraction and analysis by liquid chromatography-tandem mass spectrometry (LC-MS-MS). Analytes were separated isocratically over an 8.5-min runtime and detected on an MS-MS equipped with an electrospray ionization source operated in negative mode. A single point calibrator (15 ng/mL) forced through zero demonstrated linearity from 3 to 1,000 ng/mL. Intra- and inter-day precision were ≤9.1%, and bias was within ±14.1% for Δ8-THCCOOH and Δ9-THCCOOH. No interferences were found after challenging the method with different over-the-counter drugs, prescription pharmaceuticals, drugs of abuse and several cannabinoids and cannabinoid metabolites, including Δ10-THCCOOH. Urine specimens presumptively positive by immunoassay (n = 2,939; 50 ng/mL Δ9-THCCOOH cutoff) were confirmed with this analytical method. Δ8-THCCOOH and Δ9-THCCOOH were present together above the 15 ng/mL cutoff in 33% of specimens. However, nearly one-third of the specimens analyzed were positive for Δ8-THCCOOH only. This manuscript describes the first validated automated extraction and confirmation method for Δ8- and Δ9-THCCOOH in urine that provides adequate analyte separation in urine specimens with extreme isomer abundance ratios.

Authentic specimens of Juncaceae names stored in the Herbarium of Ivan Franko National University of Lviv

Background. The Herbarium of Ivan Franko National University of Lviv (LW) is one of the oldest herbaria in Ukraine. It was established in 1783 by Prof. B. Schiwerek. At present, its holdings are estimated at 300,000 specimens, including about 2000 type ones. Scientific processing of herbarium collections, their digitization, and cataloging are in progress. In 2002, the Herbarium of Ivan Franko National University of Lviv was included in the State Register of Scientific Objects of Ukraine that have the national heritage status. The aim of this study was to compile a list of Juncaceae type specimens of the Herbarium LW. Material and Methods. In the Herbarium LW, specimens of Juncaceae were analyzed. The authentic specimens of other herbaria were studied on JSTOR Global Plants. Using SimpleMappr, we created a map of loci classici of the type specimens names housed in this herbarium. Acronyms of herbaria are given according to the Index Herbariorum. Results. In the Herbarium of Ivan Franko National University of Lviv, we found authentic specimens of six names: Juncus bulbosus var. salinus Schur, Juncus czetzii Schur, Juncus kotschyi Boiss., Juncus lampocarpus var. viviparus Schur, Juncus transsilvanicus Schur, Luzula alpigena Schur. For each name, we provide: 1) nomenclatural citation, 2) information from the protologue, 3) the text of the herbarium label of the type specimen kept in the Herbarium LW, 4) herbarium specimen number, 5) notes. Most of the authentic specimens are kept in F. Schur’s nominative collection. Conclusions. Six authentic specimens of Juncaceae are kept in the Herbarium LW: one isolectotype and 5 syntypes. Collectors of type specimens are F. Schur and T. Kotschy. The geographic areas of the Juncaceae type collection include 2 countries: Romania and Iran. At present, all of these names belong to synonyms.

Screening and confirmation of psilocin, mitragynine, phencyclidine, ketamine and ketamine metabolites by liquid chromatography-tandem mass spectrometry.

A safe and productive workplace requires a sober workforce, free from substances that impair judgment and concentration. Although drug monitoring programs already exist, the scope and loopholes of standard workplace testing panels are well known, allowing other substances to remain a source of risk. Therefore, a high-throughput urine screening method for psilocin, mitragynine, phencyclidine, ketamine, norketamine and dehydronorketamine was developed and validated in conjunction with a urine and blood confirmation method. There are analytical challenges to overcome with psilocin and mitragynine, particularly when it comes to drug stability and unambiguous identification in authentic specimens. Screening and confirmation methods were validated according to the American National Standards Institute/Academy Standards Board (ANSI/ASB) Standard 036, Standard Practices for Method Validation in Forensic Toxicology. An automated liquid handling system equipped with dispersive pipette extraction tips was utilized for preparing screening samples, whereas an offline solid-phase extraction method was used for confirmation sample preparation. Both methods utilized liquid chromatography-tandem mass spectrometry to achieve limits of detection between 1-5 ng/mL for the screening method and 1 ng/mL for the confirmation method. Automation allows for faster throughput and enhanced quality assurance, which improves turnaround time. Compared to previous in-house methods, specimen volumes were substantially decreased for both blood and urine, which is an advantage when volume is limited. This screening technique is well suited for evaluating large numbers of specimens from those employed in safety-sensitive workforce positions. This method can be utilized by workplace drug testing, human performance and postmortem laboratories seeking robust qualitative screening and confirmation methods for analytes that have traditionally been challenging to routinely analyze.

The genus Artemisia (Asteraceae) in the historical collection of W.S.J.G. Besser (KW-BESS) at the National Herbarium of Ukraine (KW)

Our herbarium inventory revealed that the collection of specimens of the genus Artemisia (Asteraceae) in the memorial collection of W.S.J.G. Besser (KW-BESS) at the National Herbarium of Ukraine (KW) contains at least 1186 herbarium sheets. The collection of specimens of Artemisia is stored in folders Nos. 234–249 of the Besser herbarium. Each folder contains individual paper folders (here termed taxon covers). In total, there are 217 individual taxon folders. Our analysis of protologues and herbarium labels demonstrated that the KW-BESS collection contains types and/or other original specimens in 118 taxon folders, but much more actual types of species-rank and infraspecific names. Additional analysis of their possible type status is needed for specimens in 28 taxon folders. The KW-BESS collection contains authentic and historical specimens of such famous researchers and collectors of the 19th century as A. Bunge, L.K.A. Chamisso, A.P. de Candolle, D. Douglas, J.F.G. Eschscholtz, F.E.L. Fischer, J.É. Gay, A. Gray, J.D. Hooker, V.V. Jacquemont, G.S. Karelin and I.P. Kirilov, K.F. Ledebour, A.F. Marschall von Bieberstein, C.A. Meyer, T. Nuttall, J.D. Prescott, Ch. Steven, N.S. Turczaninow, N. Wallich, etc. We present and discuss here some examples of labels written by Besser and several other famous botanists. Our preliminary assessment of this globally important collection established the foundations for further nomenclatural and taxonomic studies of Artemisia, in particular, those based on historical specimens at KW.

In search of lost ergots: phylogenetic re-evaluation of Claviceps species in Japan and their biogeographic patterns revealed

Claviceps (Clavicipitaceae, Hypocreales) was erected in 1853, although ergotism had been well-known for a much longer time. By 2000, about 70 taxa had been described in Claviceps, of which eight species and six varieties were based on Japanese type or authentic specimens. Most of these Japanese Claviceps taxa are based on lost specimens or have invalid names, which means many species practically exist only in the scientific literature. The ambiguous identities of these species have hindered taxonomic resolution of the genus Claviceps. Consequently, we sought and collected more than 300 fresh specimens in search of the lost Japanese ergot. Multilocus phylogenetic analyses based on DNA sequences from LSU, TEF-1α, TUB2, Mcm7, and RPB2 revealed the phylogenetic relationships between the Japanese specimens and known Claviceps spp., as well as the presence of biogeographic patterns. Based on the phylogenetic analysis, host range and morphology, we re-evaluated Japanese Claviceps and recognised at least 21 species in Japan. Here we characterised 14 previously described taxa and designated neo-, lecto- and epi-types for C. bothriochloae, C. imperatae, C. litoralis, C. microspora, C. panicoidearum and C. yanagawaensis. Two varieties were elevated to species rank with designated neotypes, i.e. C. agropyri and C. kawatanii. Six new species, C. miscanthicola, C. oplismeni, C. palustris, C. phragmitis, C. sasae and C. tandae were proposed and described.

3D Carbonate Digital Rock Reconstruction by Self-Attention Network and GAN Structure

Amidst the rapid advancements in digital technology, the pursuit of simulating geologic and mineralogic samples in a digital domain has garnered considerable attention, becoming a linchpin in modern earth science and petrological research. This manuscript intricately explores the deployment of state-of-the-art generative models for the meticulous reconstruction of digital rock core samples. Central to this investigation was the innovative incorporation of the self-attention mechanism—a pioneering endeavor in the domain of digital rock core studies. By harnessing the prowess of this sophisticated model, we endeavored to produce samples that echo the nuanced geological and mineralogical attributes emblematic of authentic rock specimens. Distinguishing our approach, the generative architecture, bolstered by the self-attention mechanism, demonstrated unparalleled proficiency in replicating quintessential rock features, ranging from porosity and granular texture to contiguous core sequences. Additionally, the idiosyncrasies of carbonate rocks were meticulously captured, highlighting phenomena like dissolution. Empirical evaluations, rooted in stringent statistical analyses, attested to the model’s capability to generate outputs that resonate closely with genuine samples. This exploration not only amplifies the potential applications of our proposed model in geoscientific endeavors but also signals a transformative stride in digital rock physics, emphasizing the harmonious amalgamation of innovative computational models with profound geological insights.

In search of lost ergots: phylogenetic re-evaluation of Claviceps species in Japan and their biogeographic patterns revealed.

Claviceps (Clavicipitaceae, Hypocreales) was erected in 1853, although ergotism had been well-known for a much longer time. By 2000, about 70 taxa had been described in Claviceps, of which eight species and six varieties were based on Japanese type or authentic specimens. Most of these Japanese Claviceps taxa are based on lost specimens or have invalid names, which means many species practically exist only in the scientific literature. The ambiguous identities of these species have hindered taxonomic resolution of the genus Claviceps. Consequently, we sought and collected more than 300 fresh specimens in search of the lost Japanese ergots. Multilocus phylogenetic analyses based on DNA sequences from LSU, TEF-1α, TUB2, Mcm7, and RPB2 revealed the phylogenetic relationships between the Japanese specimens and known Claviceps spp., as well as the presence of biogeographic patterns. Based on the phylogenetic analysis, host range and morphology, we re-evaluated Japanese Claviceps and recognised at least 21 species in Japan. Here we characterised 14 previously described taxa and designated neo-, lecto- and epi-types for C. bothriochloae, C. imperatae, C. litoralis, C. microspora, C. panicoidearum and C. yanagawaensis. Two varieties were elevated to species rank with designated neotypes, i.e. C. agropyri and C. kawatanii. Six new species, C. miscanthicola, C. oplismeni, C. palustris, C. phragmitis, C. sasae and C. tandae were proposed and described. Taxonomic novelties: New species: Claviceps miscanthicola E. Tanaka, Claviceps oplismeni E. Tanaka, Claviceps palustris E. Tanaka, Claviceps phragmitis E. Tanaka, Claviceps sasae E. Tanaka, Claviceps tandae E. Tanaka; New status and combination: Claviceps agropyri (Tanda) E. Tanaka, Claviceps kawatanii (Tanda) E. Tanaka; Typifications (basionyms): Lecto- and epitypification: Claviceps yanagawaensis Togashi; Neotypifications: Claviceps purpurea var. agropyri Tanda, Claviceps bothriochloae Tanda & Y. Muray, Claviceps imperatae Tanda & Kawat., Claviceps microspora var. kawatanii Tanda, Claviceps litoralis Kawat., Claviceps microspora Tanda, Claviceps panicoidearum Tanda & Y. Harada; Resurrection: Claviceps queenslandica Langdon. Citation: Tanaka E, Tanada K, Hosoe T, Shrestha B, Kolařík M, Liu M (2023). In search of lost ergots: phylogenetic re-evaluation of Claviceps species in Japan and their biogeographic patterns revealed. Studies in Mycology 106: 1-39. doi: 10.3114/sim.2022.106.01.

A-049 Inter-assay Variation of Insulin-like Growth Factor-1 Remains an Issue

Abstract Background Human insulin-like growth factor-1 (IGF-1) is the main mediator of the somatotropic effects of growth hormone (GH). Accurate measurement of IGF-1 is required for the diagnosis and management of GH secretion disorders. Several commercial vendors and laboratories have standardized their IGF-1 assays to the WHO International Standard 02/254. This standardization does not prohibit the existence of significant inter-assay variability in IGF-1 measurements. Such concentration variance can lead to inaccurate patient case decision making. The IGF-1 testing performed in our laboratory was recently migrated from an Immulite 2000 platform to a CobasPro system. This shift in vendor necessitated an investigation of the inter-assay variability between these standardized tests. This relative variance was established through a validation of the analytical performance of the CobasPro IGF-1 assay. Methods Reportable range, intra-day precision and accuracy of IGF-1 measurements using the CobasPro (Roche Diagnostics) system were determined. Patient correlation studies between the CobasPro and an Immulite 2000 (Siemens Healthcare Diagnostics) instrument was also performed. The concentration of IGF-1 in a subset of these correlation study specimens was also quantified by liquid chromatography mass spectrometry (LC-MS) assays from two different reference laboratories. Results The CobasPro assay had a verified linear range of 7–1443 ng/mL (R2 = 0.9998, slope = 0.9915). At IGF-1 concentrations of 54 and 345 ng/mL the intra-day precision (%CV, N = 10) of the CobasPro assay did not exceed 0.9%. Patient specimen correlations between the CobasPro and Immulite 2000 assays showed significant bias (Deming regression: y = 1.543x − 48.61, N = 128, R = 0.9641). This method bias was exacerbated at Immulite 2000 derived IGF-1 concentrations &amp;gt;100 ng/mL (Deming regression: y = 1.662x − 84.29, N = 93, R = 0.9492, CobasPro range = 77–910 ng/mL, Immulite 2000 range = 103–539 ng/mL). Patient specimen correlations between the CobasPro and LC-MS assay from reference laboratory A showed a positive bias (Deming regression: y = 1.375x − 64.5, N = 19, R = 0.9210). An analogous correlation with reference laboratory B demonstrated a higher level of concordance (Deming regression: y = 1.005x + 10.2, N = 16, R = 0.9499). The CobasPro and Immulite 2000 assays both produced acceptable result when challenged by multiple external quality assurance (EQA) specimens provided by the College of American Pathologists (CAP). However, the observed relative trends in IGF-1 concentrations from these challenges were opposite to our observations with authentic patient specimens. The CobasPro assay consistently produced lower IGF-1 concentrations relative to the Immulite 2000 assay for these EQA specimens. Conclusion The CobasPro IGF-1 assay offered acceptable analytical performance. Although both assays were traceable to a reference standard, a significant positive concentration bias was observed between the CobasPro and Immulite 2000 assays. Standardization efforts have not eliminated inter-assay IGF-1 variability. A careful evaluation of the impact of this variance on the diagnosis and management of GH deficiency and acromegaly must be considered when IGF-1 testing platforms are changed.

Authentic Pathology Specimen Reception: A Valuable Resource for Developing Biomedical Science Student Competencies and Employability.

Background/Introduction: The pathology specimen reception is fundamental to the services provided by Biomedical Science laboratories worldwide. To ensure patient safety and that samples are of adequate quality to send for analysis, prospective Biomedical Scientists should have a robust knowledge of the processes involved and the acceptance criteria of the pathology specimen reception. This knowledge has been highlighted by employers as a current gap in Biomedical Science graduates and therefore needs to be addressed within higher education settings. To do this, this study aimed to 1) design a practical session to simulate the key processes of the pathology specimen reception and 2) to understand Biomedical Science students' opinions on these activities and the development of transferable skills required for post-graduate employment. Methods: The practical session was designed based on industrial requirements and academic knowledge of student skill sets to ensure suitability. Qualitative information regarding participant demographics and career interests was acquired through open-answer or multiple-choice questions. Quantitative student feedback was acquired via questionnaires utilising a 5-point Likert scale (n = 77). Results: The scenario-based practical session provided students with a positive learning experience with 98.7% of participants enjoying the session, with 87.0% stating they learned a lot by completing the session. It was also identified that participants preferred this style of learning to that of conventional higher education teaching modalities with 97.4% stating they would prefer simulated employment focussed scenarios embedded into the curriculum more often. The majority of participants also thought this session was helpful for the development of their key transferrable skills including teamworking, communication, and confidence. When stratified based on demographic data, there was minimal difference between cohorts and in the majority of cases, those participants from non-traditional university entry backgrounds had a more positive experience and better transferable skill development following the completion of this style of learning experience. Conclusion: This study highlights simulation-based learning as a tool to develop core Biomedical Science knowledge, build student graduate capital, and ensure the preparedness of students for post-graduation employment.

Checklist of the genus Dendrobium Sw. (Orchidaceae) in Manipur, India

An enumeration of the genus Dendrobium Sw. (Orchidaceae) in Manipur, India was attempted. Literature review revealed the occurrence of 67 species under the genus from the state. The study was carried out through repeated field explorations in different natural forests of Manipur since 2012. Out of the total taxa reported from the state, only 42 species could be traced in the field. All these taxa are presented here with their flowering phenology and places of occurrence. Twenty-five species were untraceable in the field; however, nine species out of these 25 are represented by authentic herbarium specimens and supported by published documents. The remaining 16 species were reported only in literature, but no live plants or herbarium specimens were found. Out of the 42 species inventoried from the field, many taxa are under threat owing to habitat loss due to felling of trees, deforestation, and ‘jhum’ cultivation. So, there is an urgent need of conservation of those species through in situ and ex situ means for their sustenance. A photographic plate of some taxa is provided here which might be useful for their easy identification in the field and for taking care of their conservation. Large-scale uses of these species as cut flowers and indoor &amp; outdoor plants may be practised through micropropagation and cultivation in nurseries which may help in revenue generation for the state.

The performance of metagenomic next-generation sequencing in diagnosing pulmonary infectious diseases using authentic clinical specimens: The Illumina platform versus the Beijing Genomics Institute platform.

Introduction: Metagenomic next-generation sequencing (mNGS) has been increasingly used to detect infectious organisms and is rapidly moving from research to clinical laboratories. Presently, mNGS platforms mainly include those from Illumina and the Beijing Genomics Institute (BGI). Previous studies have reported that various sequencing platforms have similar sensitivity in detecting the reference panel that mimics clinical specimens. However, whether the Illumina and BGI platforms provide the same diagnostic performance using authentic clinical samples remains unclear. Methods: In this prospective study, we compared the performance of the Illumina and BGI platforms in detecting pulmonary pathogens. Forty-six patients with suspected pulmonary infection were enrolled in the final analysis. All patients received bronchoscopy, and the specimens collected were sent for mNGS on the two different sequencing platforms. Results: The diagnostic sensitivity of the Illumina and BGI platforms was notably higher than that of conventional examination (76.9% vs. 38.5%, p < 0.001; 82.1% vs. 38.5%, p < 0.001; respectively). The sensitivity and specificity for pulmonary infection diagnosis were not significantly different between the Illumina and BGI platforms. Furthermore, the pathogenic detection rate of the two platforms were not significantly different. Conclusion: The Illumina and BGI platforms exhibited similar diagnostic performance for pulmonary infectious diseases using clinical specimens, and both are superior to conventional examinations.

Optimization of biochip assay for illegal substances on drug abusers’ whole blood: Randox Evidence vs LC-MS/MS

ABSTRACT Immunoassay tests are used in clinical and forensic toxicology laboratories to determine illicit drug use in biological samples. Therefore, this study aims to optimize the cutoff concentrations of DOA I Plus in the blood and compare the LC-MS/MS results. 680 authentic forensic whole blood specimens with Randox Evidence DOA I Plus array were screened for drug of abuse and confirmed by LC-MS/MS. Regarding the manufacturer recommended threshold values, 139 out of 680 authentic blood samples were positive for one or more analytes, while 541 were negative. Nearly all of the 139 positive blood samples confirmed by LC-MS/MS were true positive for one or more analytes, while 522 of the 541 negative blood samples were true negative. The overall sensitivity and the specificity were 87.8% and 99.6%, respectively. THC was considered in detail, and a receiver operator characteristic curve analysis was performed to determine the optimum cutoff for THC, as it accounts for 78% of all positive results according to the manufacturer’s recommended thresholds. The optimal threshold value for THC was determined at a concentration of 23 ng/mL, while these values for other parameters were defined as recommended by the manufacturer.

Microbial degradation products of lurasidone and their significance in postmortem toxicology.

Recent research reported that lurasidone degrades in unpreserved ante-mortem human whole blood inoculated with microorganisms known to dominate postmortem blood specimens. In vitro degradation occurred at a similar rate to risperidone, known to degrade in authentic postmortem specimens until below analytical detection limits. To identify the lurasidone degradation products formed, an Agilent 6520 liquid chromatograph quadrupole-time-of-flight mass spectrometer (LC-QTOF-MS) operating in auto-MS/MS mode was used. Numerous degradation products not previously reported in prior in vitro or in vivo pharmacokinetic studies or forced degradation studies were detected. Accurate mass data, mass fragmentation data, acetylation experiments, and a proposed mechanism of degradation analogous to risperidone supports initial identification of the major degradation product as N-debenzisothiazole-lurasidone (calculated m/z [M + H]+ = 360.2646). A standard was unavailable to conclusively confirm this identification. Retrospective data analysis of postmortem cases involving lurasidone identified the presence of the major degradation product in four of six cases where lurasidone was also detected. This finding is significant for toxicology laboratories screening for this drug in postmortem casework. The major postmortem lurasidone degradation product has consequently been added to the LC-QTOF-MS drug screen at Forensic Science SA (FSSA) to indicate postmortem lurasidone degradation in authentic postmortem blood specimens and as a marker of lurasidone administration in the event lurasidone is degraded to concentrations below detection limits.

Integrative Taxonomy of Nitraria (Nitrariaceae), Description of the New Enigmatic Species and Key to All Currently Known Species.

A new species, Nitraria iliensis sp. nov., is described from the Ili basin, Almaty region, Kazakhstan. It belongs to section Nitraria ser. Sibiricae and is morphologically similar to N. sibirica Pall. An integrative taxonomic approach based on molecular, biochemical and morphological analyses, along with palynological data, was used to delimit this new species. The studied species of the genus are illustrated, and photographs of authentic specimens of the new species, as well as a distribution map of the new species and segregate taxa, are provided. Morphological characters were investigated, more important traits for identification were found, and a new key to distinguish between all species of the genus was prepared.

Taxonomic revision of kelp species with rhizome-like holdfast, Laminaria longipes Bory and Laminaria repens Ruprecht, from Russian Far Eastern seas

In this paper, we describe the historical background of one Laminaria J.V. Lamouroux species from the northern Pacific region. In the references, two different Laminaria species were recorded under the same name, Laminaria longipes Bory, due to errors in the labeling of authentic herbarium specimens, a very brief description, confusion of protologues, and the absence of type specimens. One of them, inhabiting the northern Atlantic Ocean, has rhizoids, whereas the second one from the Pacific Ocean has holdfast with rhizomes. The latter species was described by Ruprecht (1850) as Laminaria repens Ruprecht, however, due to various reasons this name was not scientifically accepted and in 19th century, this kelp species has been named differently, until the name L. longipes was universally accepted. None of its previous names were typified. We found original specimens collected in the 18th and 19th centuries that were used to describe these species, designated lectotypes and specified synonyms. We propose to return the name L. repens for the Pacific Laminaria species with rhizomes. Based on molecular- phylogenetic data of our specimens, we confirmed its validity and difference from another species with rhizomes distributed in the Pacific North America, L. sinclairii (Harvey ex JD Hooker &amp; Harvey) Farlow, Anderson &amp; Eaton. We also discuss developmental biology and ecology of L. repens from the Russian Far East, provide data on the geographical distribution of all known forms of this species and show that their areas, except for the Middle Kurile Islands, where they have been known so far, cover almost the entire distribution of this species.