Development of Low-Magnetic Susceptibility Microcoils via 5-Axis Machining for Analysis of Biological and Environmental Samples.

In environmental research, it is critical to understand how toxins impact invertebrate eggs and egg banks, which, due to their tiny size, are very challenging to study by conventional nuclear magnetic resonance (NMR) spectroscopy. Microcoil technology has been extensively utilized to enhance the mass-sensitivity of NMR. In a previous study, 5-axis computer numerical control (CNC) micromilling (shown to be a viable alternative to traditional microcoil production methods) was used to create a prototype copper slotted-tube resonator (STR). Despite the excellent limit of detection (LOD) of the resonator, the quality of the line shape was very poor due to the magnetic susceptibility of the copper resonator itself. This is best solved using magnetic susceptibility-matched materials. In this study, approaches are investigated that improve the susceptibility while retaining the versatility of coil milling. One method involves machining STRs from various copper/aluminum alloys, while the other involves machining ones from an aluminum 2011 alloy and electroplating them with copper. In all cases, combining copper and aluminum to produce resonators resulted in improved line shape and SNR compared to pure copper resonators due to their reduced magnetic susceptibility. However, the copper-plated aluminum resonators showed optimal performance from the devices tested. The enhanced LOD of these STRs allowed for the first 1H-13C heteronuclear multiple quantum coherence (HMQC) of a single intact 13C-labeled Daphnia magna egg (∼4 µg total biomass). This is a key step toward future screening programs that aim to elucidate the toxic processes in aquatic eggs.

Exploring the Applications of Carbon-Detected NMR in Living and Dead Organisms Using a 13C-Optimized Comprehensive Multiphase NMR Probe.

Comprehensive multiphase-nuclear magnetic resonance (CMP-NMR) is a non-invasive approach designed to observe all phases (solutions, gels, and solids) in intact samples using a single NMR probe. Studies of dead and living organisms are important to understand processes ranging from biological growth to environmental stress. Historically, such studies have utilized 1H-based phase editing for the detection of soluble/swollen components and 1H-detected 2D NMR for metabolite assignments/screening. However, living organisms require slow spinning rates (∼500 Hz) to increase survivability, but at such low speeds, complications from water sidebands and spectral overlap from the modest chemical shift window (∼0-10 ppm) make 1H NMR challenging. Here, a novel 13C-optimized E-Free magic angle spinning CMP probe is applied to study all phases in ex vivo and in vivo samples. This probe consists of a two-coil design, with an inner single-tuned 13C coil providing a 113% increase in 13C sensitivity relative to a traditional multichannel single-CMP coil design. For organisms with a large biomass (∼0.1 g) like the Ganges River sprat (ex vivo), 13C-detected full spectral editing and 13C-detected heteronuclear correlation (HETCOR) can be performed at natural abundance. Unfortunately, for a single living shrimp (∼2 mg), 13C enrichment was still required, but 13C-detected HETCOR shows superior data relative to heteronuclear single-quantum coherence at low spinning speeds (due to complications from water sidebands in the latter). The probe is equipped with automatic-tuning-matching and is compatible with automated gradient shimming─a key step toward conducting multiphase screening of dead and living organisms under automation in the near future.

Evaluation of double-tuned single-sided planar microcoils for the analysis of small 13 C enriched biological samples using 1 H-13 C 2D heteronuclear correlation NMR spectroscopy.

Microcoils provide a cost-effective approach to improve detection limits for mass-limited samples. Single-sided planar microcoils are advantageous in comparison to volume coils, in that the sample can simply be placed on top. However, the considerable drawback is that the RF field that is produced by the coil decreases with distance from the coil surface, which potentially limits more complex multi-pulse NMR pulse sequences. Unfortunately, 1 H NMR alone is not very informative for intact biological samples due to line broadening caused by magnetic susceptibility distortions, and 1 H-13 C 2D NMR correlations are required to provide the additional spectral dispersion for metabolic assignments in vivo or in situ. To our knowledge, double-tuned single-sided microcoils have not been applied for the 2D 1 H-13 C analysis of intact 13 C enriched biological samples. Questions include the following: Can 1 H-13 C 2D NMR be performed on single-sided planar microcoils? If so, do they still hold sensitivity advantages over conventional 5 mm NMR technology for mass limited samples? Here, 2D 1 H-13 C HSQC, HMQC, and HETCOR variants were compared and then applied to 13 C enriched broccoli seeds and Daphnia magna (water fleas). Compared to 5 mm NMR probes, the microcoils showed a sixfold improvement in mass sensitivity (albeit only for a small localized region) and allowed for the identification of metabolites in a single intact D. magna for the first time. Single-sided planar microcoils show practical benefit for 1 H-13 C NMR of intact biological samples, if localized information within ~0.7 mm of the 1 mm I.D. planar microcoil surface is of specific interest.

Comprehensive Multiphase NMR Probehead with Reduced Radiofrequency Heating Improves the Analysis of Living Organisms and Heat-Sensitive Samples.

Comprehensive multiphase (CMP) NMR, first described in 2012, combines all of the hardware components necessary to analyze all phases (solid, gel, and solution) in samples in their natural state. In combination with spectral editing experiments, it can fully differentiate phases and study the transfer of chemical species across and between phases, providing unprecedented molecular-level information in unaltered natural systems. However, many natural samples, such as swollen soils, plants, and small organisms, contain water, salts, and ionic compounds, making them electrically lossy and susceptible to RF heating, especially when using high-strength RF fields required to select the solid domains. While dedicated reduced-heating probes have been developed for solid-state NMR, to date, all CMP-NMR probes have been based on solenoid designs, which can lead to problematic sample heating. Here, a new prototype CMP probe was developed, incorporating a loop gap resonator (LGR) for decoupling. Temperature increases are monitored in salt solutions analogous to those in small aquatic organisms and then tested in vivo on Hyalella azteca (freshwater shrimp). In the standard CMP probe (solenoid), 80% of organisms died within 4 h under high-power decoupling, while in the LGR design, all organisms survived the entire test period of 12 h. The LGR design reduced heating by a factor of ∼3, which allowed 100 kHz decoupling to be applied to salty samples with generally ≤10 °C sample heating. In addition to expanding the potential for in vivo research, the ability to apply uncompromised high-power decoupling could be beneficial for multiphase samples containing true crystalline solids that require the strongest possible decoupling fields for optimal detection.

Expanding current applications and permitting the analysis of larger intact samples by means of a 7 mm CMP-NMR probe.

Comprehensive multiphase NMR combines the ability to study and differentiate all phases (solids, gels, and liquids) using a single NMR probe. The general goal of CMP-NMR is to study intact environmental and biological samples to better understand conformation, organization, association, and transfer between and across phases/interfaces that may be lost with conventional sample preparation such as drying or solubilization. To date, all CMP-NMR studies have used 4 mm probes and rotors. Here, a larger 7 mm probehead is introduced which provides ∼3 times the volume and ∼2.4 times the signal over a 4 mm version. This offers two main advantages: (1) the additional biomass reduces experiment time, making 13C detection at natural abundance more feasible; (2) it allows the analysis of larger samples that cannot fit within a 4 mm rotor. Chicken heart tissue and Hyalella azteca (freshwater shrimp) are used to demonstrate that phase-based spectral editing works with 7 mm rotors and that the additional biomass from the larger volumes allows detection with 13C at natural abundance. Additionally, a whole pomegranate seed berry (aril) and an intact softgel capsule of hydroxyzine hydrochloride are used to demonstrate the analysis of samples too large to fit inside a conventional 4 mm CMP probe. The 7 mm version introduced here extends the range of applications and sample types that can be studied and is recommended when 4 mm CMP probes cannot provide adequate signal-to-noise (S/N), or intact samples are simply too big for 4 mm rotors.

5-Axis CNC Micromilling for Rapid, Cheap, and Background-Free NMR Microcoils.

The superior mass sensitivity of microcoil technology in nuclear magnetic resonance (NMR) spectroscopy provides potential for the analysis of extremely small-mass-limited samples such as eggs, cells, and tiny organisms. For optimal performance and efficiency, the size of the microcoil should be tailored to the size of the mass-limited sample of interest, which can be costly as mass-limited samples come in many shapes and sizes. Therefore, rapid and economic microcoil production methods are needed. One method with great potential is 5-axis computer numerical control (CNC) micromilling, commonly used in the jewelry industry. Most CNC milling machines are designed to process larger objects and commonly have a precision of >25 µm (making the machining of common spiral microcoils, for example, impossible). Here, a 5-axis MiRA6 CNC milling machine, specifically designed for the jewelry industry, with a 0.3 µm precision was used to produce working planar microcoils, microstrips, and novel microsensor designs, with some tested on the NMR in less than 24 h after the start of the design process. Sample wells could be built into the microsensor and could be machined at the same time as the sensors themselves, in some cases leaving a sheet of Teflon as thin as 10 µm between the sample and the sensor. This provides the freedom to produce a wide array of designs and demonstrates 5-axis CNC micromilling as a versatile tool for the rapid prototyping of NMR microsensors. This approach allowed the experimental optimization of a prototype microstrip for the analysis of two intact adult Daphnia magna organisms. In addition, a 3D volume slotted-tube resonator was produced that allowed for 2D 1H-13C NMR of D. magna neonates and exhibited 1H sensitivity (nLODω600 = 1.49 nmol s1/2) close to that of double strip lines, which themselves offer the best compromise between concentration and mass sensitivity published to date.

Spinal cord neural network interactions: implications for sympathetic control of the porcine heart.

Inherent and acquired factors determine the integrated autonomic response to cardiovascular stressors. Excessive sympathoexcitation to ischemic stress is a major contributor to the potential for sudden cardiac death. To define fundamental aspects of cardiac-related autonomic neural network interactions within the thoracic cord, specifically as related to modulating sympathetic preganglionic (SPN) neural activity. Adult, anesthetized Yorkshire pigs (n = 10) were implanted with penetrating high-density microarrays (64 electrodes) at the T2 level of the thoracic spinal cord to record extracellular potentials concurrently from left-sided dorsal horn (DH) and SPN neurons. Electrical stimulation of the T2 paravertebral chain allowed for antidromic identification of SPNs located in the intermediolateral cell column (57 of total 1,760 recorded neurons). Cardiac stressors included epicardial touch, occlusion of great vessels to transiently alter preload/afterload, and transient occlusion of the left anterior descending coronary artery (LAD). Spatial/temporal assessment of network interactions was characterized by cross-correlation analysis. While some DH neurons responded solely to changes in preload/afterload (8.5 ± 1.9%) or ischemic stress (10.5 ± 3.9%), the majority of cardiovascular-related DH neurons were multimodal (30.2 ± 4.7%) with ischemia sensitivity being one of the modalities (26.1 ± 4.7%). The sympathoexcitation associated with transient LAD occlusion was associated with increased correlations from baseline within DH neurons (2.43 ± 0.61 to 7.30 ± 1.84%, P = 0.04) and between SPN to DH neurons (1.32 ± 0.78 to 7.24 ± 1.84%, P = 0.02). DH to SPN network correlations were reduced during great vessel occlusion. In conclusion, increased intrasegmental network coherence within the thoracic spinal cord contributes to myocardial ischemia-induced sympathoexcitation.NEW & NOTEWORTHY In an in vivo pig model, we demonstrate using novel high-resolution neural electrode arrays that increased intrasegmental network coherence within the thoracic spinal cord contributes to myocardial ischemia-induced sympathoexcitation.

Ex vivo Comprehensive Multiphase NMR of whole organisms: A complementary tool to in vivo NMR.

Nuclear Magnetic Resonance (NMR) spectroscopy is a non-invasive analytical technique which allows for the study of intact samples. Comprehensive Multiphase NMR (CMP-NMR) combines techniques and hardware from solution state and solid state NMR to allow for the holistic analysis of all phases (i.e. solutions, gels and solids) in unaltered samples. This study is the first to apply CMP-NMR to deceased, intact organisms and uses 13C enriched Daphnia magna (water fleas) as an example. D. magna are commonly used model organisms for environmental toxicology studies. As primary consumers, they are responsible for the transfer of nutrients across trophic levels, and a decline in their population can potentially impact the entire freshwater aquatic ecosystem. Though in vivo research is the ultimate tool to understand an organism's most biologically relevant state, studies are limited by conditions (i.e. oxygen requirements, limited experiment time and reduced spinning speed) required to keep the organisms alive, which can negatively impact the quality of the data collected. In comparison, ex vivo CMP-NMR is beneficial in that; organisms do not need oxygen (eliminating air holes in rotor caps and subsequent evaporation); samples can be spun faster, leading to improved spectral resolution; more biomass per sample can be analyzed; and experiments can be run for longer. In turn, higher quality ex vivo NMR, can provide more comprehensive NMR assignments, which in many cases could be transferred to better understand less resolved in vivo signals. This manuscript is divided into three sections: 1) multiphase spectral editing techniques, 2) detailed metabolic assignments of 2D NMR of 13C enriched D. magna and 3) multiphase biological changes over different life stages, ages and generations of D. magna. In summary, ex vivo CMP-NMR proves to be a very powerful approach to study whole organisms in a comprehensive manner and should provide very complementary information to in vivo based research.

New records of coral-associated crabs (Decapoda: Brachyura: Carpilioidea, Trapezoidea) from Easter Island.

A review is made of those brachyurans that are symbiotic or otherwise associated with scleractinian corals on Easter Island, southeastern Pacific Ocean. A total of seven species is reported, including three species from two families not previously known from the island. Earlier records of Trapezia are analyzed and, although as many as six species have been previously reported, we conclude that only three species are known to occur on the island with certainty.

Revision of the family Chasmocarcinidae Serène, 1964 (Crustacea, Brachyura, Goneplacoidea).

The family Chasmocarcinidae Serène, 1964, is revised based on the examination of the type material of many of its species as well as unidentified and previously identified material from around the world. The revised family now consists of three subfamilies comprising 16 genera (including eight described as new) and 51 species (including 19 described as new). The subfamily Chasmocarciinae Serène, 1964, consists of Amboplax n. gen. with one species; Angustopelta n. gen. with four species, two of which are new; Camatopsis Alcock & Anderson, 1899, with six species, five of which are new; Chasmocarcinops Alcock, 1900, with one species; Chasmocarcinus Rathbun, 1898, with 11 species, one of which is new; Chinommatia n. gen. with five species, two of which are new; Deltopelta n. gen. with one species; Hephthopelta Alcock, 1899, with two species, one of which is new; Microtopsis Komai, Ng & Yamada, 2012, with two species, one of which is new; Notopelta n. gen. with one species; Statommatia n. gen. with five species, two of which are new; and Tenagopelta n. gen. with three species, two of which are new. The subfamily Megaesthesiinae Stevcic, 2005, consists of Alainthesius n. gen. with two species, both of which are new; Megaesthesius Rathbun, 1909, with four species, one of which is new. The subfamily Trogloplacinae Guinot, 1986, consists of Australocarcinus Davie, 1988, with three species, and Trogloplax Guinot, 1986, with one species. A neotype is selected for Chasmocarcinus cylindricus Rathbun, 1901. Three nominal species were found to be junior subjective synonyms of other species: Chasmocarcinus panamensis Serène, 1964, of C. longipes Garth, 1940; Chasmocarcinus rathbuni Bouvier, 1917, of C. typicus Rathbun, 1898; and Hephthopelta superba Boone, 1927, of Deltopelta obliqua (Rathbun, 1898). Thirteen chasmocarcinid genera are exclusively found in the Indo-West Pacific region, one (Chasmocarcinus) in both the Western Atlantic and Tropical Eastern Pacific regions, and two (Deltopelta n. gen. and Amboplax n. gen.) exclusively in the Western Atlantic. Chasmocarcinids are remarkable for occurring from depths exceeding 1000 m to shallow water and completely freshwater habitats: chasmocarcinines and megaesthesiines are found from shallow to deep water marine ecosystems, whereas trogloplacines live in freshwater streams, including cave systems.

New species of Latreillopsis Henderson, 1888 (Brachyura: Homolidae) and Neopalicus Moosa & Serène, 1981 (Brachyura: Palicidae) from the Hawaiian Islands.

Two new species of brachyuran crabs belonging to Latreillopsis Henderson, 1888 (Homolidae) and Neopalicus Moosa & Serène, 1981 (Palicidae) respectively are described from Maui, Hawai'i. The new species of Latreillopsis is distinguished from its nine congeners by a granular carapace and pereopods, a triangular G1, and by the distinctive ornamentation of its carapace and third maxillipeds; the new species of Neopalicus from its three congeners by the presence of three triangular anterolateral teeth, absence of extensions on the outer margins of the P3 and P4 propodi, dentate inner margins of the P3, P4 dactyli, and absence of ridges on the female abdomen. Also listed is Latreillia metanesa Williams, 1982 (Latreilliidae), recorded for the first time from the archipelago since its description from Albatross material collected in 1902.

Significance of the sexual openings and supplementary structures on the phylogeny of brachyuran crabs (Crustacea, Decapoda, Brachyura), with new nomina for higher-ranked podotreme taxa.

The patterns of complexity of the male and female sexual openings in Brachyura, which have been the source of uncertainties and conflicting opinions, are documented, together with a study of the morphologies of the coxal and sternal gonopores in both sexes, penises, spermathecae, and gonopods. The vulvae, male gonopores and penises are described among selected taxa of Eubrachyura, and their function and evolution examined in the context of a wide variety of mating behaviours. The location of female and male gonopores, the condition of the penis (coxal and sternal openings and modalities of protection), and related configurations of thoracic sternites 7 and 8, which are modified by the intercalation of a wide sternal part (thoracic sternites 7 and 8) during carcinisation, show evidence of deep homology. They represent taxonomic criteria at all ranks of the family-series and may be used to test lineages. Of particular significance are the consequences of the posterior expansion of the thoracic sternum, which influences the condition, shape, and sclerotisation of the penis, and its emergence from coxal (heterotreme) to coxo-sternal, which is actually still coxal (heterotreme), in contrast to a sternal emergence (thoracotreme). The heterotreme-thoracotreme distinction results from two different trajectories of the vas deferens and its ejaculatory duct via the P5 coxa (Heterotremata) or through the thoracic sternum (Thoracotremata). Dissections of males of several families have demonstrated that this major difference not only affects the external surface (perforation of the coxa or the sternum by the ejaculatory duct) but also the internal anatomy. There is no evidence for an ejaculatory duct passing through the articular membrane between the P5 coxa and the thoracic sternum in any Brachyura, even when the sternal male gonopore is very close to the P5 coxa. Trends towards the coxo-sternal condition are exemplified by multistate characters, varying from a shallow depression to a long groove along expanded sternites 7 and 8, and ultimately their complete, extended junction typifying the most derived coxo-sternal condition. The coxo-sternal condition is indicative of a long evolutionary history, as evidenced by the presence of multistate characters (e.g., Dorippidae, Goneplacoidea) or by a single, well-established condition (e.g., Chasmocarcinidae, Ethusidae, Panopeidae Eucratopsinae, Rhizopidae, Scalopidiidae). The penial area proves to be an essential diagnostic feature in Brachyura, with a value comparable to that of the gonopods. Penis protection is ubiquitous in Brachyura irrespective of length, and several modalities of protection prevail, which necessitate different modifications of associated structures. A long penis in a gutter developed from a partial invagination of sternite 8 induces the formation of a new "suture" at the same level as the preceding suture 6/7. Such a "supplementary suture 7/8" exists among unrelated heterotreme families (e.g., Ethusidae, Panopeidae Eucratopsinae, Pseudorhombilidae, Rhizopidae). A fully protected penis, concealed in a groove within a complete invagination of sternite 8 in the form of two contiguous plates, evolved independently (homoplasy) in Palicoidea and Chasmocarcinidae (Goneplacoidea), with sternite 8 present as a single plate in females. In condylar protection, described for the first time and occurring in several heterotreme families, the penis emerges from the extremity of the P5 coxo-sternal condyle or from its anterior border instead of from the coxa itself. A penis precisely lodged in a small excavation on sternite 8, which is lined by a row of stiff setae, is unique to Brachyura, and represents a new synapomorphy of the Homoloidea. Five modalities of penis protection are recognised in Podotremata, eight in Eubrachyura (six in Heterotremata and two in Thoracotremata). Special attention has been paid to Dorippoidea (Dorippidae and Ethusidae), which shows transformation series from coxal to coxo-sternal conditions. The coxo-sternal condition is not an intermediate towards the thoracotreme organisation, and a step in heterotreme evolution is the adoption of the coxo-sternal condition. An extreme carcinisition may also result in the sternal arrangement of male gonopores in the form of a "sternitreme" disposition, as in the case of Hymenosomatoidea, which displays a broad thoracic sternum and true sternal male gonopores (as in thoracotremes) together with several plesiomorphic traits that are assumed to represent an old, deeply-rooted heterotreme clade. A sternitreme condition evolved independently in the most ancestral heterotreme clades (such as Hymenosomatoidea) and in Thoracotremata. The older the lineage of a heterotreme is, the higher the possibility of having evolved carcinisation. Evidence that "derived" traits may be the consequence of a strong carcinisation, rather than being recently acquired, necessitates reconsidering certain character states in Brachyura. Eubrachyurans can only evolve either the heterotreme or the thoracotreme arrangement, the consistency of the inferred ancestral characters states providing a useful criterion for evaluating ancestral trait reconstructions. A widened thoracic sternum together with sternal gonopores may be present in carcinised heterotremes such as hymenosomatoids. The thoracic sternum provides a reliable complex of characters that must be carefully interpreted. The hypothesis of a coxo-sternal disposition in Cryptochiroidea and Pinnotheroidea, generally considered thoracotremes, is rejected, and an alternative interpretation of their status is discussed. A new interpretation of the phylogeny of Cryptochiroidea is outlined, but the origin of Pinnotheroidea remains puzzling. The sella turcica, frequently regarded a synapomorphy of Eubrachyura, is redefined as the structure formed by the endosternal intertagmal phragma that connects the tagma/thorax and the tagma/abdomen to thoracic interosternite 7/8. It is here termed the "brachyuran sella turcica" and is shown to be synapomorphic to all Brachyura. The Eubrachyura synapomorphically shares the fusion of the thoracic interopleurite 7/8 with the brachyuran sella turcica, forming the "eubrachyuran sella turcica". In contrast, some Podotremata (Cyclodorippoidea and Raninoidea) share a connection between the sella turcica and the thoracic interosternite 6/7. Six main patterns of the thoracic sternum in relation to variations in sutures 4/5-7/8 are recognised in Eubrachyura, whereas several subpatterns that include variations in the median line are distinguished. The evolution of the thoracic sternum and axial skeleton is reassessed in Podotremata and Eubrachyura. A posteriormost location of the male gonopore (coxal or sternal) in relation to sternite 8 characterises many brachyurans (Cryptochiroidea, Hymenosomatoidea, Majoidea, Matutidae, Menippidae, Orithyioidea, Parthenopoidea, Ucididae, Grapsoidea--including Percnidae, Plagusiidae, Varunidae), in contrast to a location close to suture 7/8 in other groups. The thoracic sternum/pterygostome junction, which has multistate characters, is shown to be a valuable taxonomic criterion. The shapes of the sterno-abdominal depression and sterno-abdominal cavity provide diagnostic features that are helpful in suprageneric assignments. The monophyly of Brachyura, Eubrachyura, and Thoracotremata is reaffirmed. The monophyly of Brachyura is supported by the interdependence of the two pairs of gonopods and penis. An abdomen permanently flexed and held by the pereopods and/or the homoloid press button (on sternite 4) or typical eubrachyuran press-button (on sternite 5) may be considered a synapomorphy of Brachyura, the absence of this condition considered a loss. The double abdominal-locking system ("double peg") on sternite 5, a device discovered in three families of the extinct Palaeocorystoidea from the Upper Aptian, is similar to the double hook present in living lyreidids, although it is lost in all other raninoid extant members. New evidence shows that the abdominal holding was an early occurrence for a brachyuran crab. The Raninoidea, sister to Palaeocorystoidea, is characterised by gymnopleurity, a condition that results from the lifting of the carapace and thus the exposure of several pleurites. The narrowing of the body and thoracic sternum, almost certainly associated with their burrowing behaviour, is a diagnostic feature of raninoid evolution, in contrast to the widening observed in the remaining Brachyura. The monophyly of Heterotremata is discussed. Although the correct assignment of the coxal male gonopore and sternal female gonopore (vulva) at the base of Decapoda and Eubrachyura, respectively, left no synapomorphies to support the Heterotremata, the group nevertheless should be regarded as the sister group to Thoracotremata. The controversial monophyly of Podotremata is discussed and arguments are presented against the suppression of this taxon. The distinction of Homoloidia from Dromioidia is argued, and a classification of Podotremata, which considers the fossil record whenever possible, is presented. The earliest brachyurans are re-examined, and a new interpretation of the phylogeny of several basal eubrachyuran groups (Dorippoidea, Inachoididae, Palicoidea, Retroplumoidea) is proposed. Stenorhynchus shares a number of characters with the Inachoididae that differentiate them from Inachidae, and also has some distinctive features that warrants its assignment to a separate inachoidid subfamily, Stenorhynchinae, which is resurrected. The concealment strategies among Brachyura are documented and discussed. Podotremes use carrying behaviour, often combined with burying and concealment under substrates, whereas living within a host, burying, and decoration are used by heterotremes, burrowing being essentially a thoracotreme strategy.(ABSTRACT TRUNCATED)

On the genus Scalopidia Stimpson, 1858 (Crustacea: Brachyura: Goneplacoidea: Scalopidiidae), with the description of one new genus and three new species.

A revision of Scalopidia Stimpson, 1858 (Brachyura: Goneplacoidea: Scalopidiidae) has resulted in the description of two new species of Scalopidia from the Indian Ocean and Papua New Guinea, as well as a new genus and new species from Madagascar. The type species of Scalopidia, S. spinosipes Stimpson, 1858, is redescribed and Hypophthalmus leuchochirus Richters, in Lenz & Richters, 1881, is synonymised with S. spinosipes.

Solid state deuteron relaxation time anisotropy measured with multiple echo acquisition.

The signal to noise ratio of solid state deuteron NMR line shapes can be significantly improved by recording multiple echoes, generated either by a quadrupole Carr-Purcell-Meiboom-Gill pulse train (QCPMG) or by magic angle spinning (MAS). It is shown in this article, theoretically and experimentally, that when these techniques are used to record partially relaxed spectra, the relaxation times of Zeeman order, T(1Z), and quadrupole order, T(1Q), measured for individual side bands in QCPMG experiments preserve relaxation time anisotropy, while rotational side bands in MAS spectra do not. The relaxation times of individual QCPMG sidebands are not identical to those measured at the same frequencies on partially relaxed quadrupole echo powder patterns, and must be computed by explicit simulation.

Mosquito production in stormwater treatment devices in the Lake Tahoe Basin, California.

In response to increasing evidence of mosquito production in structural stormwater Best Management Practices (BMPs), a collaborative project was developed to document the occurrence, species composition, and seasonal abundance of mosquitoes from selected urban and highway BMPs in the Lake Tahoe Basin, California. Structural and environmental factors associated with mosquito production in highway BMPs were identified and analyzed. Ten species of mosquitoes were collected from 47 BMPs, including Culex tarsalis, Culiseta incidens, Cs. inornata, and 7 species of Aedes. In and around South Lake Tahoe, immatures were most abundant in urban BMPs during the warmer summer and fall months, whereas natural water sources in the surrounding area harbored mosquitoes more often during the colder months of early spring. In BMPs installed along Lake Tahoe's perimeter highways, mosquitoes were observed in 11% of site visits conducted during a single season. Larval presence in highway BMPs was positively associated with water temperature and negatively associated with precipitation, sand, and unspecified organic matter. The significance of mosquito production in BMPs of the Tahoe Basin and the potential for increased transmission of mosquito-borne disease are discussed.

High field 207Pb spin-lattice relaxation in solid lead nitrate and lead molybdate.

Spin-lattice relaxation rates of lead have been measured at 17.6 T (156.9 MHz) as a function of temperature in polycrystalline lead nitrate and lead molybdate. Comparing the results with relaxation rates measured at lower fields, it is found that at high fields and low temperature, chemical shift anisotropy (CSA) makes small but observable contributions to lead relaxation in both materials. At 17.6 T and 200 K, CSA accounts for about 15% of the observed relaxation rate. Above 300 K, the dominant relaxation mechanism even at 17.6 T is an indirect Raman process involving modulation of the (207)Pb spin-rotation tensor, as first proposed by Grutzner et al. [J. Am. Chem. Soc. 123, 7094 (2001)] and later treated theoretically in more detail by Vega et al. [Phys. Rev. B 74, 214420 (2006)]. The improved signal to noise ratio at high fields makes it possible to quantify relaxation time anisotropy by analyzing saturation-recovery functions for individual frequencies on the powder pattern line shape. No orientation dependence is found for the spin-lattice relaxation rate of either material. It is argued from examination of the appropriate theoretical expressions, derived here for the first time, that the lack of observable relaxation time anisotropy is probably a general feature of this indirect Raman mechanism.

A large-scale mutagenesis screen to identify seizure-resistant zebrafish.

PURPOSE: Zebrafish are a vertebrate organism ideally suited to mutagenesis screening strategies. Although a genetic basis for seizure susceptibility and epilepsy is well established, no efforts have been made to study seizure resistance. Here we describe a novel strategy to isolate seizure-resistant zebrafish mutants from a large-scale mutagenesis screen. METHODS: Seizures were induced with pentylenetetrazole (PTZ). Zebrafish were analyzed between 3 and 7 days postfertilization (dpf). Genome mutations were induced in founders by using N-ethyl-nitrosourea (ENU). Seizure behavior was monitored by using a high-speed camera and quantified by locomotion-tracking software. Electrographic activity was monitored by using a field-recording electrode placed in the optic tectum of agar-immobilized zebrafish. RESULTS: Short-term PTZ exposure elicited a burst-suppression seizure pattern in 3-dpf zebrafish and more complex activity consisting of interictal- and ictal-like discharges at 7 dpf. Prolonged exposure to PTZ induced status epilepticus-like seizure activity and fatality in wild-type zebrafish larvae. With a PTZ survival assay at 6-7 dpf, we identified six zebrafish mutants in a forward-genetic screen covering nearly 2,000 F(2) families. One mutant (s334) also was shown to exhibit reduced behavioral activity on short-term PTZ exposure and an inability to generate long-duration ictal-like discharge. CONCLUSIONS: Zebrafish offers a powerful tool for the identification and study of a genetic basis for seizure resistance.

Hinged molecular capsules: synthesis and conformational control via temperature, pH, or solvent composition.

Three new covalently linked molecular capsules were synthesized from their resorcinarene cavitand precursors in good yields. The capsules undergo reversible conformational switching between the closed "vase" form and the open "kite" form upon temperature or pH variation. The kite conformation obtained via either method in CDCl(3) switches to vase conformation upon addition of polar solvents such as acetone-d(6) or THF-d(8).

Quantitative analysis of left atrial function during left ventricular ischemia with and without left atrial ischemia: a real-time 3-dimensional echocardiographic study.

Left atrial (LA) systolic and diastolic function were analyzed in an animal model of acute left ventricular ischemia with LA ischemia by proximal left circumflex coronary artery occlusion or without LA ischemia by midleft anterior descending coronary artery occlusion (7 sheep in each group). LA pressures and LA volumes were simultaneously recorded using a catheter and real-time 3-dimensional echocardiography, respectively. LA stroke volume represented 63% of left ventricular stroke volume during left anterior descending coronary artery occlusion, but only 32% during left circumflex coronary artery occlusion. This animal study with 3-dimensional echocardiographic determination of absolute LA volumes demonstrated dramatic depression of LA function during proximal left circumflex coronary artery occlusion as compared with left anterior descending coronary artery occlusion.