Authentic Specimens Research Articles

Medicinal plants used with Thai Traditional Medicine in modern healthcare services: A case study in Kabchoeng Hospital, Surin Province, Thailand

Ethnopharmacological relevanceThai Traditional Medicine (TTM) is available in many modern hospitals in Thailand. However, there have been difficulties in integrating TTM, particularly the practices of the use of herbal medicines, into modern healthcare services. Kabchoeng Hospital is one hospital that has been able to overcome these difficulties. Thus, this study aimed to document the successful utilization of herbal medicine at Kabchoeng Hospital. The documentation focused on both the knowledge of medicinal plants and the success factors that facilitated the utilization of herbal medicine in the context of a modern hospital in Thailand. Materials and methodsKabchoeng Hospital was intentionally selected for this case study. Participatory observation was used for the data collection. There were six groups of key informants: three applied Thai Traditional Medicine practitioners (ATTMPs), a pharmacist, two physicians, two folk healers, the head of an herbal cultivation and collection group, and 190 patients. The plant specimens were collected and identified based on the botanical literature and a comparison with authentic specimens; these identifications were assisted by microscopic and thin layer chromatography (TLC) techniques. ResultsEighty-nine medicinal plants were used for the herbal preparations. The ATTMPs used these plants to prepare 29 standard herbal preparations and occasional extemporaneous preparations. Moreover, in this hospital, seven herbal preparations were purchased from herbal medicine manufacturers. In total, 36 preparations were used for 10 groups of symptoms, such as the treatment of respiratory system disorders, musculo-skeletal system disorders, and digestive system disorders. Four success factors that facilitated the utilization of herbal medicine at Kabchoeng Hospital were determined. These factors included a proper understanding of the uses of herbal medicines, the successful integration of the modern and TTM healthcare teams, the support of an herbal cultivation and collection group, and the acceptance of the local people. ConclusionsThe practices that support the use of herbal medicine at Kabchoeng Hospital illustrated the successful application of TTM and also represented a model for the integration of TTM, and particularly the use of herbal medicine, into modern hospitals. This integration will be beneficial for sustainable healthcare systems in Thailand and in other countries where modern medicine is the mainstream medical system.

Immunoassay Screening of Diphenhydramine (Benadryl(R)) in Urine and Blood Using a Newly Developed Assay

Diphenhydramine (DPH) is a common over the counter antihistamine that produces drowsiness and has the potential to cause driving under the influence of drugs-related accidents. To date there are no commercially available immunoassay screening kits for its detection in biological fluids such as urine and/or blood. We describe a newly developed enzyme-linked immunosorbent assay (ELISA) screen and report on its utility in the analysis of authentic specimens taken from volunteers. The assay is specific for detection of DPH and does not detect closely related antihistamines like brompheniramine, chlorpheniramine, and doxylamine. There is a varying amount of cross-reactivity seen with certain tricyclic compounds, due to similarities in side chain structure with DPH. Intra- and interday precision of the assay were determined to be less than 10%. The assay is highly sensitive and has a working range from 1 to 500 ng/mL for urine and 1 to 250 ng/mL for blood. The assay was further validated with authentic urine and blood specimens obtained from volunteers and coroner's laboratories.

The Detection of 1-Palmitoyl-2-oleoyl-<i>sn</i>-glycero-3-phosphoethanol and Ethyl Glucuronide in Human Umbilical Cord

In utero exposure to ethanol continues to be a significant public health issue and neonatal healthcare professionals are in need of objective means to identify exposed newborns. The aim of this study was to fully validate two methods for the detection of two direct alcohol biomarkers, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanol (POPE) and ethyl glucuronide (EtG), in umbilical cord and apply the assays to a group of authentic specimens. The limits of detections were 2 and 1 ng/g for POPE and ETG and the limits of quantitation were 4 and 3 ng/g, respectively. Inter and intra-day precision and accuracy measurements were within 15%. The assays were applied to 308 authentic specimens where we detected POPE in five (1.6%) specimens and EtG in twelve (3.9%) specimens. The mean concentrations were 11.4 ng/g ± 9.4 ng/g and 127.2 ± 227.7 ng/g for POPE and EtG, respectively. This study suggested that umbilical cord was a suitable specimen type for the identification of newborns exposed to ethanol in the womb and the prevalence of POPE and EtG detected in umbilical cord were consistent with the prevalence of self-reported binge drinking reported by the National Birth Defect Prevention Study (NBDPS) and Behavioral Risk Factor Surveillance System (BRFSS). Further studies are required to fully describe the association between the observed concentrations of POPE and EtG in umbilical cord to the level of maternal consumption of ethanol.

Using Museums to Teach Undergraduate Paleontology and Evolution

Museum exhibitions possess a long history of serving as useful tools for teaching both paleontology and evolutionary biology to college undergraduates. Yet, they are frequently under-appreciated and underutilized. However, they remain potentially outstanding resources because they can be used to meet a spectrum of learning objectives related to nature of science, real-world relevance, and student interest. Specifically, even small museum displays can provide: 1) authentic specimens, which often are more diverse, of higher quality, and historically more significant than those in teaching collections; 2) specimens in context, with other specimens and/or geological or biological background available; 3) examples of how fossils connect to virtually all of Earth and life sciences (explaining why they have so frequently been at the center of traditional “natural history”); 4) cross-disciplinary experiences, connecting science, art, technology, and history within a social context; and 5) opportunities for students to learn about teaching. A survey of instructor-developed activities performed within a host of natural history museums—with particular attention devoted to the Museum of the Earth, an affiliate of Cornell University—suggests that natural history exhibitions, regardless of size and scope, can complement and strengthen formal education in an undergraduate setting.

Reassessment of type specimens of <i>Cordyceps</i> and its allies, described by Dr. Yosio Kobayasi and preserved in the mycological herbarium of the National Museum of Nature and Science (TNS). Part 3: <i>Cordyceps</i> s. l. on Cicadidae

Twenty taxa of Cordyceps s. l. (belonging to three genera in the latest taxonomic system) from Cicadidae had been described by Kobayasi and Shimizu. Among them, 13 holotype specimens of Cordyceps were rediscovered from preserved specimens partially without ordering, and their registration numbers (TNS-F-number) were given: viz. (1) Elaphocordyceps inegoensis TNS-F-230289, (2) Elaphocordyceps paradoxa TNS-F-230313, (3) Elaphocordyceps toriharamontana TNS-F-230288, (4) Ophiocordyceps heteropoda TNS-F-230294, (5) Ophiocordyceps longissima TNS-F-230285, (6) Ophiocordyceps prolifica f. terminalis TNS-F-230295, (7)Ophiocordyceps pseudolongissimaTNSF- 197983, (8) Ophiocordyceps takaoensis TNS-F-3026, (9) Cordyceps kanzashiana TNS-F-198015, (10) Cordyceps kobayasii Koval’ (≡C. sinclairii Kobayasi, not C. sinclairii Berk.) TNS-F-212384, (11) Cordyceps minuta TNS-F- 11933, (12) Cordyceps ramosopulvinata TNS-F-197979, and (13) Cordyceps ryogamimontana TNS-F-230292. Two specimens were selected as lectotype: viz. (14) Ophiocordyceps prolifica TNS-F-230300 and (15) Ophiocordyceps yakusimensis TNS-F-230287. A paratype specimen of (16) Cordyceps pleuricapitata TNS-F-197965, and an authentic specimen of (17) Cordyceps imagamiana TNS-F- 197966, were also rediscovered.

A comparison of two FDA approved lamotrigine immunoassays with ultra-high performance liquid chromatography tandem mass spectrometry

BackgroundLamotrigine is an anti-epileptic drug used as adjunct therapy for seizures. Lamotrigine is commonly used in pregnant women with epilepsy, a population in which therapeutic drug monitoring (TDM) is useful to optimize dose. Drug–drug interactions that can induce or inhibit metabolism or elimination and impaired hepatic function are also possible indications for lamotrigine TDM. Chromatographic techniques are currently used for performing most TDM of lamotrigine, but this may change, as automated immunoassays were recently introduced. MethodsImmunoassays available through Seradyn and ARK Diagnostics were validated using a Beckman AU400e automated chemistry analyzer. The intra-day precision was accessed with 5 replicates of three quality control materials, and inter-assay precision was estimated by assaying the same material over 4days. Linearity was evaluated by serially diluting a spiked sample and measuring it in duplicate. The 2 methods were compared with ultra high performance liquid chromatography tandem mass spectrometry (UPLC–MS/MS) using 44 authentic patient specimens. ResultsThe intra-day (n=5) and inter-assay (n=20) coefficients of variation were ≤7.5% for the 3 levels tested. The analytical measurement ranges were confirmed as stated by the manufacturers (0 or 1–40μg/ml). The percent recovery of the quality control materials and Deming regression for the 44 patient results showed good agreement of both immunoassays when compared to the UPLC–MS/MS assay. ConclusionThe lamotrigine assays studied here produced a slightly lower result than UPLC–MS/MS but were precise and easy to perform.

Infrared identification of barbiturates with particular reference to the occurrence of polymorphism.

Abstract Infrared spectra of twelve substituted barbituric acids, in a total of 34 polymorphic forms, have been compared. Comparison of a sample spectrum with that of an authentic specimen provides a reliable means of identification, provided that both are in the same crystalline form. To ensure consistent production of a single form a specific treatment is recommended for each substance exhibiting polymorphism.

Infrared identification of pharmaceutically important sulphonamides with particular reference to the occurrence of polymorphism.

Abstract The infrared absorption spectra of sulphonamides, when compared with the spectra of Authentic Specimens, provide a simple and complete means of identification, provided the effects of polymorphism are excluded. Of 18 substances examined, twelve showed evidence of polymorphism. Limited solubility prevents the use of solution spectra, and specified solvent treatments, details of which are given, may therefore be necessary with these substances to ensure reproducible spectra.

INFRARED IDENTIFICATION OF PHARMACEUTICALLY IMPORTANT STEROIDS WITH PARTICULAR REFERENCE TO THE OCCURENCE OF POLYMORPHISM.

Abstract The infrared absorption spectra of steroids, when compared with the spectra of Authentic Specimens, provide a simple and complete means of identification, provided that the effects of polymorphism are precluded. Of 35 substances examined, 16 showed no evidence of polymorphism and a further twelve were sufficiently soluble to be examined in solution. Specific solvent treatments, details of which are given, may be necessary with the remaining seven substances if the spectra of the sample and of the Authentic Specimen are not identical when first examined.

The detection of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanol in human dried blood spots

Phosphatidylethanol, a series of abnormal phospholipids formed in the presence of ethanol and phospholipase D, has gained popularity as a long-term biomarker of ethanol ingestion. A liquid chromatography tandem mass spectrometric method for a specific, prevalent isomer, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanol, was developed and validated using dried blood spots. Dried blood spots offer numerous advantages over venipuncture including reduced costs, invasiveness and discomfort. Dried blood spots were prepared from authentic whole blood specimens that had been tested using a previously published procedure. Comparison of the results from the two assays demonstrated excellent correlation. The data suggest that dried blood spots may be a useful tool for the detection of alcohol abuse and abstinence monitoring.

Concentration and UV reactivity of total organic carbon in the upper layer in the oligotrophic subtropical western North Pacific

Characteristics of temporal variation in the vertical profile of total organic carbon (TOC) were investigated in the upper layer of the ocean at various latitudes (10–30°N) of 137°E in the western North Pacific subtropical gyre. Bulk TOC was divided into labile and stable fractions by the specific UV irradiation to evaluate their contributions to the temporal fluctuation of TOC. Authentic specimens and dissolved organic carbon collected from coastal waters were used to examine the correspondence between the photochemical reactivity to the specific UV irradiation and biological reactivity. The concentration of TOC in the upper layer was higher in summer when the surface layer is well stratified and was lower in winter when the mixed layer deepened. Although the winter mixed layer is shallower in lower latitudes, the amplitude of the seasonal variations in the top 50 m weighted mean TOC concentration (19.5–27.7 μM) did not vary considerably over the subtropical zones we studied. A major fraction (51.9–89.8%) of TOC was stable against specific UV irradiation and was thought to be mainly composed of biologically stable organic compounds, but the seasonal variation of TOC at each latitude was mainly attributable to the variation of UV labile fraction. This finding suggests that the seasonal variation of TOC in the upper layer is explained by the biological production of TOC in summer surface water and its vertical mixing with low UV, labile TOC in winter. Upward advection of subsurface water due to the passage of cyclonic eddies and changes in the geostrophic current are also likely to affect the temporal TOC variation, in particular, in the lower latitudes where winter mixed layer and permanent thermocline are shallower.

Characterization of in vitro generated metabolites of the selective androgen receptor modulators S‐22 and S‐23 and in vivo comparison to post‐administration canine urine specimens

Selective androgen receptor modulators (SARMs) have great therapeutic potential in various diseases including cancer cachexia, sarcopenia, and osteoporosis, and the number of drug candidates has been growing over the last decade. The SARM drug candidates S-22 and S-23 belong to one of the most advanced groups of androgen receptor modulators and are based on an arylpropionamide-derived core structure. Due to their anabolic effects, SARMs have been prohibited in elite sports and have been a subject of sports drug testing programmes since January 2008. Consequently, the structure of analytically useful urinary metabolites should be elucidated to provide targets for sensitive and retrospective analysis. In the present study, the phase-I and -II metabolism of S-22 and S-23 was simulated using hepatic human enzymes, and resulting metabolites were characterized by means of state-of-the-art mass spectrometric approaches employing high resolution/high accuracy Orbitrap mass spectrometry. Subsequently, the newly defined target compounds including the glucuronic acid conjugates of S-22 and S-23, their corresponding monohydroxylated and bishydroxylated analogs, as well as their B-ring depleted counterparts were implemented into an existing routine doping control procedure, which was examined for its specificity for the added substances. In order to obtain proof-of-concept data for authentic urine specimens, canine urine samples collected up to 72 h after oral administration of S-22 to dogs were analyzed using the established approach outlining the capability of the presented assay to detect the glucuronide of S-22 as well as the B-ring-depleted metabolite (M3) in all samples following therapeutic (31.4 µg/kg) dosing. Finally, M3 was chemically synthesized, characterized by nuclear magnetic resonance spectroscopy and high resolution/high accuracy mass spectrometry, and chosen as primary target for future doping control analyses.

Automated Solid-Phase Extraction-Liquid Chromatography-Tandem Mass Spectrometry Analysis of 6-Acetylmorphine in Human Urine Specimens: Application for a High-Throughput Urine Analysis Laboratory

An automated solid-phase extraction-liquid chromatography- tandem mass spectrometry (SPE-LC-MS-MS) method using the Spark Holland Symbiosis Pharma SPE-LC coupled to a Waters Quattro Micro MS-MS was developed for the analysis of 6-acetylmorphine (6-AM) in human urine specimens. The method was linear (R² = 0.9983) to 100 ng/mL, with no carryover at 200 ng/mL. Limits of quantification and detection were found to be 2 ng/mL. Interrun precision calculated as percent coefficient of variation (%CV) and evaluated by analyzing five specimens at 10 ng/mL over nine batches (n = 45) was 3.6%. Intrarun precision evaluated from 0 to 100 ng/mL ranged from 1.0 to 4.4%CV. Other opioids (codeine, morphine, oxycodone, oxymorphone, hydromorphone, hydrocodone, and norcodeine) did not interfere in the detection, quantification, or chromatography of 6-AM or the deuterated internal standard. The quantified values for 41 authentic human urine specimens previously found to contain 6-AM by a validated gas chromatography (GC)-MS method were compared to those obtained by the SPE-LC-MS-MS method. The SPE-LC-MS-MS procedure eliminates the human factors of specimen handling, extraction, and derivatization, thereby reducing labor costs and rework resulting from human error or technique issues. The time required for extraction and analysis was reduced by approximately 50% when compared to a validated 6-AM procedure using manual SPE and GC-MS analysis.

Tiselius'PotamogetonExsiccates: Changes in Taxonomy and Nomenclature from One-Century Perspective

Tiselius' outstanding collection Potamogetones suecici exsiccati is reviewed and re-evaluated from the viewpoint of the present nomenclature and taxonomy. All 150 gatherings issued in this collection are listed and interpreted taxonomically. In a few cases new taxonomic findings are given based on molecular investigation of plant material from the type localities. All new names proposed in this collection are listed and checked for their nomenclatural validity and legitimacy. Lectotypes are designated for 64 valid names and authentic specimens listed for 27 additional invalid designations. Two names (and their types) are of particular importance for contemporary plant taxonomy because these are the correct names for recognized and confirmed Potamogeton hybrids: P. × torssanderi for the triple hybrid P. gramineus × P. lucens × P. perfoliatus and P. × lanceolatifolius for P. gramineus × P. nodosus.

Psorantin, a unique methylene linked dimer of vismin and kenganthranol E, two anthranoid derivatives from the fruits of Psorospermum aurantiacum (Hypericaceae)

Two prenylated anthranoids, psorantin and kenganthranol E, were isolated from the fruit of Psorospermum aurantiacum together with the known compounds ferruginin B, vismin, vismion D, haronginanthrone, kenganthranol B, kenganthraquinone, 1,7-dihydroxyxanthone, paradisiol, fridelin, fridelinol and betulinic acid. Their structures were determined on the basis of spectral data and by comparison with data reported in the literature as well as with authentic specimens for the known compounds. The structures of the new compounds were determined as bis-[3,8,9-trihydroxy-6-methyl-4,4-bis-(3,3-dimethylallyl)anthracenyl]methane ( 1) and 1,8,10-trihydroxy-6-methyl-4,5-bis-(3,3-dimethylallyl)-2,3-(2,2-dimethylpyrano)anthrone ( 2). Psorantin ( 1) is a dimer of vismin formed through a methylene linkage. The two new compounds when tested against the microbial strains Bacillus megaterium, Escherichia coli, Chlorella fusca and Microbotryum violaceum showed no activity.

Determination of eight fatty acid ethyl esters in meconium samples by headspace solid‐phase microextraction and gas chromatography–mass spectrometry

A number of fatty acid ethyl esters (FAEEs) have recently been detected in meconium samples. Several of these FAEEs have been evaluated as possible biomarkers for in utero ethanol exposure. In the present study, a method was optimized and validated for the simultaneous determination of eight FAEEs (ethyl laurate, ethyl myristate, ethyl palmitate, ethyl palmitoleate, ethyl stearate, ethyl oleate, ethyl linoleate and ethyl arachidonate) in meconium samples. FAEEs were extracted by headspace solid-phase microextraction. Analyte detection and quantification were carried out using GC-MS operated in chemical ionization mode. The corresponding D5-ethyl esters were synthesized and used as internal standards. The LOQ and LOD for each analyte were <150 and <100 ng/g, respectively. The method showed good linearity (r(2)>0.98) in the concentration range studied (LOQ-2000 ng/g). The intra- and interday imprecision, given by the RSD of the method, was lower than 15% for all FAEEs studied. The validated method was applied to 63 authentic specimens. FAEEs could be detected in alcohol-exposed newborns (>600 ng/g cumulative concentration). Interestingly, FAEEs could also be detected in some non-exposed newborns, although the concentrations were much lower than those measured in exposed cases.

Reclassification of Two Peronospora Species Parasitic on Draba in Hyaloperonospora Based on Morphological and Molecular Phylogenetic Data

On the family Brassicaceae, the causal agent responsible for downy mildew disease was originally regarded as a single species, Peronospora parasitica (now under Hyaloperonospora), but it was recently reconsidered to consist of many distinct species. In this study, 11 specimens of Peronospora drabae and P. norvegica parasitic on the genus Draba were investigated morphologically and molecularly. Pronounced differences in conidial sizes (P. drabae: 14-20×12.5-15.5μm; P. norvegica: 20-29×15.5-22μm) and 7.8% sequence distance between their ITS1-5.8S-ITS2 rDNA sequences confirmed their status as distinct species. Based on ITS phylogeny and morphology (monopodially branching conidiophores, flexuous to sigmoid ultimate branchlets, hyaline conidia and lobate haustoria), the two species unequivocally belong to the genus Hyaloperonospora and not to Peronospora to which they were previously assigned. Therefore, two new combinations, Hyaloperonospora drabae and H. norvegica, are proposed. The two taxa are illustrated and compared using the type specimen for H. norvegica and authentic specimens for H. drabae, which is lectotypified.

Effect of hydrolysis on identifying prenatal cannabis exposure

Identification of prenatal cannabis exposure is important due to potential cognitive and behavioral consequences. A two-dimensional gas chromatography-mass spectrometry method for cannabinol, Delta(9)-tetrahydrocannabinol (THC), 11-hydroxy-THC (11-OH-THC), 8beta,11-dihydroxy-THC, and 11-nor-9-carboxy-THC (THCCOOH) quantification in human meconium was developed and validated. Alkaline, enzymatic, and enzyme-alkaline tandem hydrolysis conditions were optimized with THC- and THCCOOH-glucuronide reference standards. Limits of quantification ranged from 10 to 15 ng/g, and calibration curves were linear to 500 ng/g. Bias and intra-day and inter-day imprecision were <12.3%. Hydrolysis efficiencies were analyte-dependent; THC-glucuronide was effectively cleaved by enzyme, but not base. Conversely, THCCOOH-glucuronide was most sensitive to alkaline hydrolysis. Enzyme-alkaline tandem hydrolysis maximized efficiency for both glucuronides. Identification of cannabinoid-positive meconium specimens nearly doubled following alkaline and enzyme-alkaline hydrolysis. Although no 11-OH-THC glucuronide standard is available, enzymatic hydrolysis improved 11-OH-THC detection in authentic specimens. Maximal identification of cannabis-exposed neonates and the widest range of cannabis biomarkers are achieved with enzyme-alkaline tandem hydrolysis.

Politamide, a new constituent from the stem bark of Ficus polita Vahl (Moraceae)

A new cerebroside 1 characterized as (2R,9Z)-2-hydroxy-N-{(1S,2S,3R,4S)-1-[(β-D-glucopyranosyloxy)methyl]-2,3,4-trihydroxyoctacosan-1-yl}-9-pentadecenamide was isolated from the stem bark of Ficus polita Vahl (Moraceae) together with four known compounds identified as sitosterol 3-O-β-D-glucopyranoside 2, betulinic acid 3, stigmasterol 4 and lupeol 5. Their structures were determined on the basis of spectroscopic methods as well as HR-ESI-MS, NMR analyses, chemical transformation, and by comparison of their physical and spectral data with those reported in the literature and with authentic specimens for some known compounds.

Identity and neotypification of &lt;I&gt;Craterellus cinereus&lt;/I&gt; and description of &lt;I&gt;Cantharellus atrofuscus&lt;/I&gt; sp. nov.

The study of authentic specimens of Cantharellus cinereus, present in the Leiden Herbarium (L), has allowed us to ascertain the identity of this species, which has no clamp connections. There exists a species close to Craterellus (Cantharellus) cinereus distinguished by important macroand microscopic features, including an unperforated basidioma and presence of clamp connections. This species is here proposed as new to science with the name Cantharellus atrofuscus. A description and photos of habit and microscopic characters are provided.