Settling strategy of stalked echinoderms from the Kaili Biota (middle Cambrian), Guizhou Province,South China

A total of 124 articulated echinoderms, including Sinoeocrinus lui (n = 30), “S. globus” (n = 92), one indeterminate gogiid, and one indeterminate edrioasteroid, from the Kaili Formation (Middle Cambrian), eastern Guizhou, China was examined. Among them, 73% of gogiid echinoderms are preserved attached to skeletal substrates. Suitable skeletal substrates for gogiid holdfasts in the Kaili Biota are organophosphatic brachiopods; large trilobite fragments, including cranidia, free cheeks, thoracic segments, and pygidia; hyoliths; and Scenella shells. The high frequency of “S. globus” attached to organophosphatic brachiopods is due to the high richness of organophosphatic brachiopods rather than a host-specific association in the Kaili Biota. However, the possible discrimination against attachment to small trilobites, such as Pagetia, requires further investigation. Based on 11 hand samples, echinoderm population density (EPD) varies from 0.3 to 4.5 echinoderms per 50 grind points (0.06 to 0.96 echinoderms per cm²), depending on the percent coverage of skeletal debris. Crowding effect is evident when EPD is approximately 0.32 echinoderms per cm² and the skeletal coverage is less than 20%.     

Multiple 2D approaches to human sexual dimorphism of the distal end of femur

Various studies on the difference of distal femoral condyles between genders have been reported recently in order to provide anatomic information for knee prosthesis design and surgical planning in total knee arthroplasty.They also had the objective to be used as a sex recognition character,as may be needed in forensic medicine.Except for a recent 3D approach on the distal femur,most of the studies used dimensional information or aspect ratio but not shape.Our 2D study aimed to determine the size and shape variation of femoral condyles in Thais,considering age,sex and sides.One hundred and twenty-four cadaveric femurs (male 84 legs and female 40 legs) were dissected.The specimens were photographed by digital camera and images were analyzed using three geometric techniques:(i) the landmark-based method (5 landmarks),(ii) with or without addition of 23 sliding semilandmark and (iii) the outline-based methods.From the resulting geometric coordinates,size and shape were extracted for comparisons between genders and sides.Between sides,directional asymmetry could be detected only for shape variation,and only when introducing curves in the analyses (either through the semilandmarks technique or through the outline-based one).Non-directional asymmetry,probably fluctuating asymmetry,was detected for size,as well as for shape,in both genders.Sex discrimination was performed for each geometric technique using two classification methods:the Mahalanobis distance classification and the Maximum likelihood classification.The latter provided much more satisfactory gender validated reclassification (87％) than shape (72％).     

Principles for the virtual reconstruction of hominin crania

Fossils are usually discovered broken or distorted, therefore reconstruction is inevitably the first step towards any comparative analysis. We outline a general methodological framework by which missing information about biological specimens can be estimated using geometric morphometric methods and discuss how this relates to effective paleoanthropological use of incomplete and distorted crania.Combining digital data resources with geometric morphometrics, we go beyond the assembly of fragments on the computer. As in a three-dimensional jigsaw puzzle, we first assemble the virtual pieces manually. Then we use landmarks, several hundred semilandmarks, and information from complete specimens to estimate missing coordinates and correct for distortion simultaneously. One can thus incorporate information from incomplete specimens in a comparative morphometric analysis while keeping track of the uncertainties that result from partial preservation or deformation. We exemplify our approach by reconstructing the fossil crania Arago XXI, Taung, and KNM-WT 15000. As different assumptions and algorithms lead to different estimations, there exists no “all-purpose” reconstruction. Instead one creates multiple reconstructions—a posterior distribution in a Bayesian sense. This distribution reflects uncertainty due to missing data values and sensitivity to prior assumptions. While there will typically be shape differences among equally plausible reconstructions, these different estimates might still support a single conclusion.     

Crypto-helical body plan in partially disarticulated gogiids from the Cambrian of South China

All living echinoderms have a pentaradial symmetry that is unique within the Bilateria. However, the Cambrian origin of echinoderm radial/pentaradiate symmetry is a long-standing problem. During the Cambrian (542–488 Ma), gogiids were the most common stalked echinoderm characterized by an “irregularly” plated body. Based on recently discovered material from the Balang Formation (Cambrian Series 2), eastern Guizhou, China, three unusual, partially disarticulated specimens of Guizhoueocrinus have clear evidence for a helical body plan. This helical plating is only evident in partially disarticulated specimens, thus a crypto-helical body construction is present. Crypto-helical construction in a gogiid raises the possibility of a phylogenetic connection among helicoplacoids, gogiids, and Helicocystis. The crypto-helical body construction may be an important evolutionary innovation among pre-radiate echinoderms.     

Geometric and traditional morphometrics for the assessment of character state identity: multivariate statistical analyses of character variation in the genus Arrenurus (Acari,Hydrachnidia, Arrenuridae)

Geometric and traditional morphometric approaches are tested to describe and reveal taxonomic characters and character states in variation of shape and size of idiosoma morphology in the Arrenuridae. Patterns of variation of idiosoma and glandularia features of males from 11 Mexican species of Arrenurus (Megaluracarus) and two species of subgenus Dadayella were explored with five landmark configurations and three sets of interlandmark distances. Separate principal component analyses (PCA) and canonical variate analyses (CVA) were performed for each data set. The eight multivariate analyses of variance among 13 a priori groups (species) detected significantly different morphometric variants, which were interpreted as different taxonomic character states. Patterns of character state similarity among species were examined with unweighted‐pair grouping method using averages (UPGMA) cluster analyses on Mahalanobis distances. Analyses of five landmark configurations revealed important taxonomical variation in the anterior idiosoma outline (10 character states), the outline of the posterior region or cauda (13 states), the distribution of postocularia, and the second and third pairs of dorsoglandularia (nine states), the fourth pair of dorsoglandularia (three states), and ventroglandularia on the posterior side of idiosoma (nine character states). Multivariate analyses of three sets of distance measurements also resulted in the detection of potential taxonomic characters related to idiosoma size (12 character states), postocularia and dorsoglandularia (13 states), and ventroglandularia (nine character states). Morphometric analyses of distances and shapes provide a formal basis for the interpretation of taxonomic characters, and for the discovery of character states. These characters should be investigated further in a wider sample of species for the phylogenetic systematics of these water mites. In the meantime, idiosoma regions and structures were tested for congruence in a phylogenetic analysis, and were proposed as homologous among the species sampled.     

Geometric morphometrics on Greek house mouse populations (Mus musculus domesticus) with Robertsonian and all-acrocentric chromosomal arrangements

This work aims to give the first comprehensive morphometric analysis of intraspecific variation for the different populations of the western house mouse (Mus musculus domesticus), in the Robertsonian (Rb) system of the N-NW Peloponnisos. Furthermore, we study all-acrocentric karyotype populations (2n = 40) of the species coming from several localities of Greece. We apply 2D shape analysis, i.e. landmark analysis and Elliptic Fourier Analysis, on the dorsal and ventral side of skull and the occlusal view of the first upper molar (M¹), respectively. Although significant genetic divergence between typical and Rb populations and even ongoing speciation processes have been reported for this species, this was not the case for the Greek populations studied. However, our analyses herein reveal morphologically differentiated chromosome groups in N-NW Peloponnisos Rb system and a clear geographical discrimination of the all-acrocentric (2n = 40) populations for all characters studied. We suggest that in all-acrocentric (2n = 40) karyotype mice the geographical distance drives their differentiation while within the Rb system of N-NW Peloponnisos, karyotype is the key factor that acts on their phenotypic variation.     

Analyse et modélisation statistique linéaire de la structure osseuse crânienne à partir de données morphométriques et anthropométriques pour la prédiction de paramètres mécaniques

The aim of this study is to establish and quantify the relationship between quasi-static stiffness and anthropometric and morphometric parameters of cranial osseous bones. The purpose is to predict the fracture parameters of cranial bone in crashworthiness situation. Within an experimental protocol, 289 specimens are taken from 17 unembalmed human skulls; the subjects are aged between 53 and 82. The specimen are tested under quasi-static three-points bending tests, at 10 mm/min. Morphometric data are got by analysing specimens’ photography. Values are studied by statistical analysis based on linear methods. Data are resumed by Component Principal Analysis. Groups of specimen, in function of the area of extraction of the specimen, are detected. The relationship between the stiffness and the morphometric parameters of the specimen is studied by using multiple linear regression; the input data are the weight, the density and the curvature indices of each specimen. The regression equation for the frontal and parietal areas is confirmed by residuals study; the relative error between predicted values and observed values is 3 ± 25%. There is no linear regression for the specimen taken from the temporal area of the skull.     

First mitochondrial DNA analysis of the spectacled porpoise (Phocoena dioptrica) from Tierra del Fuego,Argentina

The spectacled porpoise (Phocoena dioptrica, Lahille, 1912) is one of the cetacean species about which least is known. Most information on the biology and ecology of this species has been obtained from stranded specimens and sightings at sea. In this study, analysis of 380 bp fragment of mitochondrial DNA (mtDNA) control region sequences (N = 50) was performed to provide a preliminary assessment of the genetic variation in spectacled porpoise specimens found stranded or by caught on the coast of Tierra del Fuego, Argentina. Results showed high levels of mtDNA diversity, as expected in large size and stable populations, and similar to other species of porpoises. The star-like shape phylogeny of haplotypes indicates a recent population expansion. This is the first report on the genetic variation of this species. Other lines of evidence (microsatellite loci, single-nucleotide polymorphism (SNPs)) are needed to improve our knowledge on the molecular biology of the spectacled porpoise.     

Sensitive detection of histamine using fluorescently labeled oxido-reductases

A detection scheme is described by which the histamine contents of biological samples can be established. The scheme is based on the use of methylamine dehydrogenase (MADH) which converts primary amines into the corresponding aldehydes and ammonia. The generated reducing equivalents are subsequently transferred to the physiological partner of MADH, amicyanin, which thereby is converted from the oxidized blue-colored form into the reduced colorless form. The change in absorption is detected by monitoring the fluorescence of a covalently attached Cy5 dye label whose fluorescence is (partly) quenched by Förster resonance energy transfer (FRET) to the Cu-site of the amicyanin. The quenching efficiency and, thereby, the label fluorescence, depends on the oxidation state of the amicyanin. When adding histamine to the assay mixture the proportionality between the substrate concentration and the observed rate of the fluorescence increase has enabled this assay as a sensor method with high sensitivity. The MADH and amicyanin composition can be tuned so that the sensor can be adapted over a broad range of histamine concentrations (13 nM–225 μM). The lowest concentration detected so far is 13 nM of histamine. The sensor retained its linearity up to 225 μM with a coefficient of variation of 11% for 10 measurements of 100 nM histamine in a 100 μL sample volume. The use of a label fluorescing around 660 nm helps circumventing the interference from background fluorescence in biological samples. The sensor has been tested to detect histamine in biological fluids such as fish extracts and blood serum.     

A geometric morphometric analysis of the shape of the first upper molar in mice of the genus Mus (Muridae, Rodentia)

Phenotypic variation in the shape of the first upper molar among 595 mice, representing nine extant and three extinct taxa of the genus Mus , was studied with thin-plate spline analysis. The reliability of classification of individual specimens into known groups based on their molars varied from 75 to 100%, depending on group and method used. Including 13 sliding semilandmarks to the analysis improved the detection of different kinds of size and shape variation as well as visualization of shape differences between studied groups. Correlation between phylogenetic and morphometric distances suggested about 80% contribution of phylogenetic inertia to the molar shape variation; moreover, the importance of localized versus global shape changes was similar in the detection of phylogenetic signals. Finally, shape changes along individual evolutionary lineages were revealed, suggesting a few cases of reversals, convergence and/or retention of ancestral shape. The evolution of mouse molars has thus been driven by random effects of drift together with stabilizing selection and convergence.     

Glycomic analyses of ovarian follicles during development and atresia

To examine the detailed composition of glycosaminoglycans during bovine ovarian follicular development and atresia, the specialized stromal theca layers were separated from the stratified epithelial granulosa cells of healthy (n = 6) and atretic (n = 6) follicles in each of three size ranges: small (3–5 mm), medium (6-9 mm) and large (10 mm or more) (n = 29 animals). Fluorophore-assisted carbohydrate electrophoresis analyses (on a per cell basis) and immunohistochemistry (n = 14) were undertaken. We identified the major disaccharides in thecal layers and the membrana granulosa as chondroitin sulfate-derived ?uronic acid with 4-sulfated N-acetylgalactosamine and ?uronic acid with 6-sulfated N-acetylgalactosamine and the heparan sulfate-derived Δuronic acid with N-acetlyglucosamine, with elevated levels in the thecal layers. Increasing follicle size and atresia was associated with increased levels of some disaccharides. We concluded that versican contains 4-sulfated N-acetylgalactosamine and it is the predominant 4-sulfated N-acetylgalactosamine proteoglycan in antral follicles. At least one other non- or 6-sulfated N-acetylgalactosamine proteoglycan(s), which is not decorin or an inter-α-trypsin inhibitor family member, is present in bovine antral follicles and associated with hitherto unknown groups of cells around some larger blood vessels. These areas stained positively for chondroitin/dermatan sulfate epitopes [antibodies 7D4, 3C5, and 4C3], similar to stem cell niches observed in other tissues. The sulfation pattern of heparan sulfate glycosaminoglycans appears uniform across follicles of different sizes and in healthy and atretic follicles. The heparan sulfate products detected in the follicles are likely to be associated with perlecan, collagen XVIII or betaglycan.     

Phylomineralogy of the Coralline red algae: Correlation of skeletal mineralogy with molecular phylogeny

The coralline algae in the orders Corallinales and Sporolithales (subclass Corallinophycidae), with their high degree of mineralogical variability, pose a challenge to projections regarding mineralogy and response to ocean acidification. Here we relate skeletal carbonate mineralogy to a well-established phylogenetic framework and draw inferences about the effects of future changes in sea-water chemistry on these calcified red algae. A collection of 191 coralline algal specimens from New Zealand, representing 13 genera and 28 species, included members of three families: Corallinaceae, Hapalidiaceae, and Sporolithaceae. While most skeletal specimens were entirely calcitic (range: 73–100 wt.% calcite, mean 97 wt.% calcite, std dev = 5, n = 172), a considerable number contained at least some aragonite. Mg in calcite ranged from 10.5 to 16.4 wt.% MgCO₃, with a mean of 13.1 wt.% MgCO₃ (std dev = 1.1, n = 172). The genera Mesophyllum and Lithophyllum were especially variable. Growth habit, too, was related to mineralogy: geniculate coralline algae do not generally contain any aragonite. Mg content varied among coralline families: the Corallinaceae had the highest Mg content, followed by the Sporolithaceae and the Hapalidiaceae. Despite the significant differences among families, variation and overlap prevent the use of carbonate mineralogy as a taxonomic character in the coralline algae. Latitude (as a proxy for water temperature) had only a slight relationship to Mg content in coralline algae, contrary to trends observed in other biomineralising taxa. Temperate magnesium calcites, like those produced by coralline algae, are particularly vulnerable to ocean acidification. Changes in biomineralisation or species distribution may occur over the next few decades, particularly to species producing high-Mg calcite, as pH and CO₂ dynamics change in coastal temperate oceans.     

A three‐dimensional geometric morphometric study of the development of sulcus versus shell outline in Permian neospiriferine brachiopods

The morphological variation of the sulcal development and shell outline in large Permian neospiriferine brachiopods including Fasciculatia Waterhouse, 2004 is investigated using geometric morphometrics. The sulcal tongues of spiriferide brachiopods can be, in a qualitative sense, categorized into three types according to the degree of their development: short sulcal tongue, long sulcal tongue and geniculated sulcal tongue. All three types have been noted within Fasciculatia striatoparadoxa, regardless of the nature of the substrate which they originally inhabited. To quantify its morphological variation both in sulcal development and shell outline, 51 brachiopod shells were scanned with a three‐dimensional (3‐D) surface imaging device, and their 3‐D models were reconstructed. Using two landmarks and 58 semilandmarks designated on the surface of the reconstructed 3‐D models, a landmark‐based morphometric analysis was performed. Our result demonstrates a significant intraspecific variation of sulcal development in F. striatoparadoxa and its relatives. Local environmental factors, especially the intensity of ambient water flow, are invoked as the most likely cause for this intraspecific variation. Additionally, this study also shows that there are considerable interspecific distinctions in shell outline among Fasciculatia species, independent of the high variation in the sulcal development. The strong stability of overall shell outline at species level implies a decoupled morphological development between sulcal tongue and whole shell outline. The 3‐D morphometric approach applied here demonstrates its great utility as a tool for quantifying and analysing the morphological variation of highly convex brachiopod shells.     

Ecological gradients and environmental impact in the forest dwelling Haplothrips subtilissimus (Thysanoptera: Phlaeothripidae) phenotypic variability

Classification of phenotypic variability in forest dwelling arthropods, especially in applied bioindication, remains difficult due to the complex and synergic ecological interactions. Ecologically, it corresponds with the concept of alternative ontogenies in order to maintain high population fitness. The research on the geobiont Haplothrips subtilissimus (Thysanoptera: Phlaeothripidae) morphometric plasticity alongside selected ecological gradients (temperature, humidity, food availability) and the environmental impact in the xerothermous hilly oak wood (Martinský les, SW Slovakia), have revealed that (MAN(C)OVA and Kruskal–Wallis test) the head width (p < 0.001) and anterior pronotum width (p ≤ 0.002) as the only statistically significant variables. The Kruskal–Wallis test (non parametric ANOVA) revealed significant variation (more noticeable in males) in ecologically more diverse ecotone stands. A significant correlation (p < 0.001) could be shown between the morphometric parameter pairs “head width” and “anterior pronotum width” for both sexes (0.729 for females and 0.822 for males) and between anterior and posterior pronotum width only for males (0.859). Pillai's statistics revealed an interaction of soil humidity and soil temperature which confirms female body parameters depend on habitat conditions (p = 0.011). Females responded mainly to soil temperature (p = 0.018) and food availability (p = 0.030). Soil humidity interacts with the male's morphology (soil humidity p = 0.040), while the relationship to food availability (Acarina) was not statistically significant (p = 0.350). No body parameter has been significantly affected by environmental stress.     

Purification and characterisation of alliinase produced by Cupriavidus necator and its application for generation of cytotoxic agent: Allicin

Allicin, an extremely active constituent of freshly crushed garlic, is produced upon reaction of alliin with the enzyme alliinase (EC 4.4.1.4). A bacterium Cupriavidus necator with the ability of alliinase production was isolated from a soil sample and was identified by morphological, biochemical and 16S rRNA sequence. Alliinase production was optimised and it was further purified to apparent homogeneity with 103-fold purification and specific activity of 209 U/mg of protein by using DEAE Cellulose and Sephadex G-100 chromatography. The enzyme is a homodimer of molecular weight 110 kDa with two subunits of molecular weight 55 kDa each. The optimum activity of the purified enzyme was found at pH 7 and the optimum temperature was 35 °C. The enzyme exhibited maximum reaction rate (V_max) at 74.65 U/mg and Michaelis–Menten constant (K_m) was determined to be 0.83 mM when alliin was used as a substrate. The cytotoxic activity of in-situ generated allicin using purified alliinase and alliin was assessed on MIA PaCa-2 cell line using MTT assay and Acridine orange–ethidium bromide staining. This approach of in-situ allicin generation suggests a novel therapeutic strategy wherein alliin and alliinase work together synergistically to produce cytotoxic agent allicin.     

Revisiting historical Khoe-San skeletal remains in European collections: A search for identity through craniometric analysis

As the identity of a large number of Khoe-San skeletal material in European collections recently came into question during its documentation, a re-evaluation of the remains by employing a non-invasive method such as craniometrics was done to investigate the biological affinity. For this purpose, gene flow and population diversity present within the group, as well as between the study sample (N = 63) and other modern and historic population groups from southern, central and east Africa were explored. Available comparative groups included the historic Khoe-San from Riet River (N = 31), the Sotho-Tswana from southern Africa (N = 61), the Basuku from central Africa (N = 66) and the Bahutu (N = 53) and Teita (N = 24) from east Africa. Ten craniometric variables were selected and used to perform population structure analysis based on model bound quantitative genetics and multiple discriminant function analysis (MDA). Quantitative genetic distances revealed that the Khoe-San sample was closest to the Riet River group. Residual variance analysis performed on two-sample subsets of the Khoe-San group (Cape KS and Various KS) showed a higher level of heterogeneity in the Cape KS than seen in the Khoe-San from various other areas in southern Africa. MDA revealed that Khoe-San intra-sample variance is relatively high, with 44% of the sample (sexes pooled) classified into the Riet River group. The remaining individuals were classified (in decreasing order) into Bahutu (24%), Basuku (24%) and Sotho-Tswana (8%). Although the Khoe-San specimens are closest to the Riet River group, they are clearly not homogenous. Their high level of phenotypic diversity most likely originated from a complex population history involving many group interactions driven by social and political marginalization.     

Objective assessment of goat sperm head size by computer-assisted sperm morphometry analysis (ASMA)

The aim of the present study was to identify different objective categories of sperm head size evaluation, using both morphometric and statistical analyses. For this purpose, semen samples (n = 16) were collected from 4 Florida male goats and assessed using a computer-assisted sperm morphometric analysis (ASMA) to obtain sperm head size measurements. The sperm heads were grouped into categories according to 25th and 75th percentiles (the lower and upper quartile, respectively) of their area values. Thereafter, a discriminant analysis was implemented on all the sperm morphometric parameters assessed, to obtain a classification matrix for sperm head size. Sperm heads by this method were classified into 3 categories depending on their size (small, medium and large), with a globally correct assignment of 95.5%. Moreover, significant differences (p < 0.001) were recorded between individual animals for all the sperm head morphometric parameters assessed. In conclusion, by using both statistical and morphometric analyses, it was possible to recognize the three categories of sperm head size. It is expected that research could be useful in defining the relationship between sperm head size measurements and actual fertility data.     

Radiation damage effects at four specimen temperatures from 4 to 100 K

Radiation damage is the primary factor that limits resolution in electron cryo-microscopy (cryo-EM) of frozen-hydrated biological samples. Negative effects of radiation damage are attenuated by cooling specimens to cryogenic temperatures using liquid nitrogen or liquid helium. We have examined the relationship between specimen temperature and radiation damage across a broad spectrum of resolution by analyzing images of frozen-hydrated catalase crystal at four specimen temperatures: 4, 25, 42, and 100 K. For each temperature, “exposure series” were collected consisting of consecutive images of the same area of sample, each with 10 e^?/Ų exposure per image. Radiation damage effects were evaluated by examining the correlation between cumulative exposure and normalized amplitudes or IQ values of Bragg peaks across a broad range of resolution (4.0–173.5 Å). Results indicate that for sub-nanometer resolution, liquid nitrogen specimen temperature (100 K) provides the most consistent high-quality data while yielding statistically equivalent protection from radiation damage compared to the three lower temperatures. At lower resolution, suitable for tomography, intermediate temperatures (25 or 42 K) may provide a modest improvement in cryo-protection without introducing deleterious effects evident at 4 K.     

Pre-cooling with intermittent ice ingestion lowers the core temperature in a hot environment as compared with the ingestion of a single bolus

The timing in which ice is ingested may be important for optimizing its success. However, the effects of differences in the timing of ice ingestion has not been studied in resting participants. Therefore, the purpose of this study was to investigate the effects of differences in the timing of ice ingestion on rectal temperature (Tre) and rating of perceptual sensation in a hot environment. Seven males ingested 1.25 g kg⁻¹ of crushed ice (ICE1.25: 0.5 °C) or cold water (CON: 4 °C) every 5 min for 30 min, or were given 7.5 g kgBM⁻¹ of crushed ice (ICE7.5) to consume for 30 min in a hot environment (35 °C, 30% relative humidity). The participants then remained at rest for 1 h. As physiological indices, Tre, body mass and urine specific gravity were measured. Rating of thermal sensation was measured at 5-min intervals throughout the experiment. ICE1.25 continued to decrease Tre until approximately 50 min, and resulted in a greater reduction in Tre (−0.56±0.20 °C) than ICE7.5 (−0.41±0.14 °C). Tre was reduced from 40 to 75 min by ICE1.25, which is a significant reduction in comparison to ICE7.5 (p<.05). Mean RTS with ICE1.25 at 50–65 min was significantly lower than that with ICE7.5 (p<.05). These results suggest that pre-cooling with intermittent ice ingestion is a more effective strategy both for lowering the Tre and for the rating of thermal sensation.