Development of areolae and growth of the peripheral prothallus in the crustose lichen Rhizocarpon geographicum: an image analysis study

Areolae of the crustose lichen Rhizocarpon geographicum (L.) DC., are present on the peripheral prothallus (marginal areolae) and also aggregate to form confluent masses in the centre of the thallus (central areolae). To determine the relationships between these areolae and whether growth of the peripheral prothallus is dependent on the marginal areolae, the density, morphology, and size frequency distributions of marginal areolae were measured in 23 thalli of R. geographicum in north Wales, UK using image analysis (Image J). Size and morphology of central areolae were also studied across the thallus. Marginal areolae were small, punctate, and occurred in clusters scattered over the peripheral prothallus while central areolae were larger and had a lobed structure. The size-class frequency distributions of the marginal and central areolae were fitted by power-law and log-normal models respectively. In 16 out of 23 thalli, central areolae close to the outer edge were larger and had a more complex lobed morphology than those towards the thallus centre. Neither mean width nor radial growth rate (RaGR) of the peripheral prothallus were correlated with density, diameter, or area fraction of marginal areolae. The data suggest central areolae may develop from marginal areolae as follows: (1) marginal areolae develop in clusters at the periphery and fuse to form central areolae, (2) central areolae grow exponentially, and (3) crowding of central areolae results in constriction and fragmentation. In addition, growth of the peripheral prothallus may be unrelated to the marginal areolae.     

Carbohydrates in the hypothallus and areolae of the crustose lichen <Emphasis Type="Italic">Rhizocarpon geographicum</Emphasis> (L.) DC.

Carbohydrate concentrations in the marginal hypothallus and areolae of the crustose lichen Rhizocarpon geographicum (L.) DC. were measured in north Wales, U.K. using gas chromatography. Ribitol, arabitol, and mannitol were the most abundant carbohydrates while α- glucose β-glucose, fructose, sucrose, and trehalose were present in smaller amounts. The concentrations of arabitol, ribitol, mannitol, fructose, and α-glucose were greater in the areolae while the concentration of trehalose was greater in the hypothallus. Concentrations of carbohydrates varied little between sample days. Concentrations of polyols in the hypothallus were not correlated with those in the areolae. These results suggest: 1) the hypothallus has a lower demand for carbohydrates than the areolae or there is limited transport from areolae to hypothallus, 2) increased trehalose in the non-lichenised hypothallus may be an adaptation to withstand stress and desiccation, and 3) polyols are partitioned differently in the hypothallus and areolae.     

Hidradenitis suppurativa of the breast areolae. Case report.

A case of hidradenitis suppurativa of the areolae of the breasts is presented. The treatment was excision and split-skin grafting, and this was successful.     

Scanning electron microscopy of the glomerular filtration membrane in the rat kidney

The rat kidney was perfused with saline and glutaraldehyde, treated with Murakami's tannin-osmium impregnation method, ethanol-freeze cracked and dried by the critical point method. Gold-palladium evaporated specimens were observed in a field-emission scanning electron microscope. The glomerular filtration membrane, fractured in different planes was observed with the following results: 1. Adjacent pedicles originate from different podocytes. No interpedicular bridges of apparent cytoplasmic nature could be found. 2. The basement membrane, in grazing fractures shows a horizontally layered architecture. 3. The attenuated endothelial sheet (lamina fenestrata) is divided into compartments, which we suggest should be called "areolae fenestratae", by cytoplasmic crests radiating from the nucleated portion of the endothelial cell. A crest also occurs along the cell margin, which contacts a similar crest at the margin of the adjacent cell. 4. The pores in the areolae fenestratae are variable in size (30-150 nm diameter). A knob-like projection from the apparently naked basement membrane is found in a portion of the pores. 5. Numerous microvilli may occur on the endothelium. Some of them anastomose and fuse with one another to form a net whose meshes appear identical with the endothelial pores. Domes and shelves formed of a fenestrated cytoplasmic sheet also occur above the ordinary level of the endothelial lining. A hypothesis implicating microvilli in the partial renewal of the endothelial sheet is proposed.     

Opening and closing of a toroidal group II chaperonin revealed by a symmetry constrained elastic network model

Recently, the atomic structures of both the closed and open forms of Group 2 chaperonin protein Mm‐cpn were revealed through crystallography and cryo‐electron microscopy. This toroidal‐like chaperonin is composed of two eightfold rings that face back‐to‐back. To gain a computational advantage, we used a symmetry constrained elastic network model (SCENM), which requires only a repeated subunit structure and its symmetric connectivity to neighboring subunits to simulate the entire system. In the case of chaperonin, only six subunits (i.e., three from each ring) were used out of the eight subunits comprising each ring. A smooth and symmetric pathway between the open and closed conformations was generated by elastic network interpolation (ENI). To support this result, we also performed a symmetry‐constrained normal mode analysis (NMA), which revealed the intrinsic vibration features of the given structures. The NMA and ENI results for the representative single subunit were duplicated according to the symmetry pattern to reconstruct the entire assembly. To test the feasibility of the symmetry model, its results were also compared with those obtained from the full model. This study allowed the folding mechanism of chaperonin Mm‐cpn to be elucidated by SCENM in a timely manner.     

HERITABILITY OF MANTLE AREOLAR CHARACTERS IN AULACOSEIRA SUBARCTICA (BACILLARIOPHYTA)1

Are size and arrangement of valve mantle areolae in Aulacoseira Thwaites adapted to light intensity? To test one criterion demonstrating an adaptation, heritability experiments were run on isolates of Aulacoseira subarctica (Müller) Haworth. Several clones of A. subarctica were isolated from Yellowstone Lake (Wyoming, USA), Lewis Lake (Wyoming), and East Rosebud Lake (Montana, USA). Two to four clones from each lake were grown in batch cultures under three irradiance levels: 2, 11.4, and 115 μmol photons·m^?2·s^?1. Five randomly chosen valves for each of two replicates of each clone were examined using a scanning electron microscope for a total of 300 valves. Size measurements were taken for each valve examined, and images of mantle areolae were captured on film at a magnification of 20,000×. Each image was digitized, and quantitative morphometric areolar characters were measured. A quantitative genetic analysis was performed within each light environment for the mean area of the external opening of mantle areolae, the mean distance between areolae within pervalvar striae, and the mean distance between pervalvar striae. Resulting estimates of heritability from among‐lake and within‐lake analyses indicate that all three mantle areolar characters could presently respond to selection and thus have potentially done so in the past.     

A two-exposure technique for ice-embedded samples successfully reconstructs the chlorocruorin pigment of Sabella spallanzanii at 2. 1 Nm resolution.

A technique for reconstructing ice-embedded macromolecules from electron micrographs taken at two specimen tilts (+/-23 degrees ) has been used to determine the structure of chlorocruorin isolated from the Polychaete annelid Sabella spallanzanii. Images of individual molecules were extracted in couples from two micrographs of the same field of view so each couple consists of two projections of the same molecule. One couple was used as a fixed reference for alignment. Different references yielded reconstructions with different orientations. These were merged to give a model against which the orientation of 1624 first-exposure images was refined to give a final reconstruction at 2.1 nm resolution. The structure of this hematic pigment, essentially the same as that for Lumbricus terrestris, is a bilayer structure with overall symmetry D6, containing six hollow groups per layer. A hollow group is formed by six globular masses and has approximate threefold symmetry. Other structural elements connect the two layers and the hollow groups in a layer. This non-globin material occupies about 15% of the total molecular volume. The results show that the double-exposure strategy, previously described by some of the authors and tested in computer simulations, performs well in real experiments and could be used to obtain preliminary reconstructions in a semiautomatic way.     

The effect of estrogen receptor genotype on litter size and placental traits at term in F2 crossbred gilts

The effect of estrogen receptor (ESR) genotype (two alleles, A and B) on litter size of 275 Large White x Meishan F2 crossbred gilts (73 AA, 126 AB and 76 BB gilts) was tested. In addition, for 63 of these gilts (18 AA, 24 AB, and 21 BB) the effect of ESR genotype on average placental traits at term was tested, since individual placental information was available for 88% of the 628 liveborn piglets. Without affecting average birth weight of the piglets, ESR genotype significantly affected litter size, i.e. AB gilts had larger litters than BB gilts (P < 0.05). Total number born was 11.38+/-0.38, 11.88+/-0.28, and 10.68+/-0.35, while number born alive was 10.45+/-0.39, 11.07+/-0.29, and 9.85+/-0.36 for AA, AB and BB gilts, respectively. Since the B allele in previous research was associated with largest litters, the hypothesis that ESR is a marker rather than the major gene itself is discussed. Average placental length, surface area, and weight including and excluding amnion were not affected by ESR genotype. However, placentae of AB gilts had a significantly lower number of areolae per placenta than BB gilts and had a lower number of areolae/cm2 placenta than AA and BB gilts. Number of areolae was 8945+/-663, 7240+/-619, and 9694+/-633, for AA, AB and BB gilts, respectively. Although the reason for the low number of areolae on placentae in AB gilts is not yet known, the results suggest that the ESR linked major gene for litter size might be involved in the development and activity of endometrial glands.     

Piglet and placental traits at term in relation to the estrogen receptor genotype in gilts

Liveborn piglets from gilts with estrogen receptor (ESR) genotype AA (95 AA-AA and 91 AA-AB piglets), AB (88 AB-AA, 118 AB-AB, and 37 AB-BB piglets), and BB (97 BB-AB and 89 BB-BB piglets) were compared after farrowing, to examine whether piglet ESR genotype (ESRp) nested within maternal ESR genotype (ESRm) affected placental traits at term, piglet birth weight, and growth until weaning. Furthermore, the relation of birth weight to various placental traits and the relations between placental traits were evaluated relative to ESR genotype. For this study, 62 Large White x Meishan F2 crossbred gilts (18 AA, 24 AB, and 20 BB) were used. The gilts belonged to a population in which the A allele is favorable for litter size. ESRp nested within ESRm did not affect placental length, weight, surface area and number of areolae. ESRp nested within ESRm affected amnion weight (AA-AA amnions were heavier than AA-AB, AB-AA and BB-AB amnions), placental weight after including placental surface area in the model (AA-AB placentae were lighter than AA-AA, AB-BB and BB-AB placentae), placental efficiency calculated as birth weight divided by placental weight (AB-AA placentae were less efficient than AA-AB placentae), and the relations of birth weight to placental weight and birth weight to number of areolae. The found differences imply an interaction of maternal and fetal ESR genotype on placental traits (especially weight and number of areolae) during fetal development. Furthermore, the found effects on placental and amnion weight might be the result of a difference in thickness or vascularization or both. The favorable ESR allele for litter size, i.e. the A allele, appears to be the unfavorable allele for pre-weaning piglet growth. Therefore, further research on ESR in relation to vascularization, weight and thickness of placentae. uterine size, endometrial gland development, and piglet growth is recommended.     

SCANNING ELECTRON MICROSCOPY OF THECAE OF THE DINOFLAGELLATE GENUS ORNITHOCERCUS1

Members of the genus Ornithocercus are all tropical oceanic species in which the theca is extended in the form of elaborate wing-like projections, the lists, supported by ribs. This paper illustrates the topography of 6 of the species. The theca is penetrated by numerous simple pores opening flush with the outer surface. On the inner side of the thecae the pores have a raised rim. The hypotheca of all species examined except O. splendidus have shallow depressions, the areolae, over most of the surface. Secondary thickening of mature cell walls deepens the appearance of the areolae, and increases their extent over the hypotheca in O. quadratus. The number of pores is not directly correlated with the areolae but seems rather to be a function of cell size. A comparison of the surface features confirms views that O. splendidus occupies a relatively isolated phenetic position. It also confirms the close similarity of O. steinii with O. thumii. Unexpectedly it suggested a phenetic closeness between O. magnificus and O. quadratus on the basis of hypothecal structure and rib features of the left sulcal list. O. heteroporus could not be subjected to the same degree of study and its position cannot be commented on. Some theoretical hydrodynamic and morphogenetic problems in Ornithocercus are discussed.     

Symmetry analysis of cell nuclei

An algorithm is described that is used to analyze the two-dimensional spatial symmetry of cell nuclei. The method provides two symmetry features: the symmetry index (SI), which estimates the precise spatial symmetry of a given chromatin component, Cn, and the quadrant symmetry index (QSI), which estimates the number of quadrants being occupied by Cn. A previous analysis is used to show that age-related change in Malpighian tubule nuclei from the adult housefly is associated with significant alterations in the spatial symmetry of low-, medium-, and high-density chromatin components (LDC, MDC, HDC). This included a seven-fold increase in the spatial symmetry of HDC and a shift in the symmetry profile (from highest to lowest degree of symmetry) from LDC-MDC-HDC to MDC-LDC-HDC. The increased spatial symmetry of HDC suggests that it occurs at new nuclear sites as the fly ages and that these sites are distributed over approximately 60% of the chromosome population.     

3D structure of Sulfolobus solfataricus carboxypeptidase developed by molecular modeling is confirmed by site-directed mutagenesis and small angle X-ray scattering

Sulfolobus solfataricus carboxypeptidase (CPSso) is a thermostable zinc-metalloenzyme with a M(r) of 43,000. Taking into account the experimentally determined zinc content of one ion per subunit, we developed two alternative 3D models, starting from the available structures of Thermoactinomyces vulgaris carboxypeptidase (Model A) and Pseudomonas carboxypeptidase G2 (Model B). The former enzyme is monomeric and has one metal ion in the active site, while the latter is dimeric and has two bound zinc ions. The two models were computed by exploiting the structural alignment of the one zinc- with the two zinc-containing active sites of the two templates, and with a threading procedure. Both computed structures resembled the respective template, with only one bound zinc with tetrahedric coordination in the active site. With these models, two different quaternary structures can be modeled: one using Model A with a hexameric symmetry, the other from Model B with a tetrameric symmetry. Mutagenesis experiments directed toward the residues putatively involved in metal chelation in either of the models disproved Model A and supported Model B, in which the metal-binding site comprises His(108), Asp(109), and His(168). We also identified Glu(142) as the acidic residue interacting with the water molecule occupying the fourth chelation site. Furthermore, the overall fold and the oligomeric structure of the molecule was validated by small angle x-ray scattering (SAXS). An ab initio original approach was used to reconstruct the shape of the CPSso in solution from the experimental curves. The results clearly support a tetrameric structure. The Monte Carlo method was then used to compare the crystallographic coordinates of the possible quaternary structures for CPSso with the SAXS profiles. The fitting procedure showed that only the model built using the Pseudomonas carboxypeptidase G2 structure as a template fitted the experimental data.     

On the morphology and taxonomy of <Emphasis Type="Italic">Parlibellus Crucicula</Emphasis> (Bacillariophyta)

A study of the phytoplankton of the water bodies and watercourses in the Yarayakha River and Naduiyakha River basins (the Yamal Peninsula) has revealed a wide morphological variability of the quantitative diagnostic features in Parlibellus crucicula: valve length and width, the number of striae and areolae in 10 μm, and the stria density in the middle part of the valve. A number of the features (raphe structure, presence of pseudosepta, and obliquely oriented oval areolae) typical for the genus Prestauroneis have been found in Parlibellus crucicula. This makes it possible to define this species as belonging to the genus Prestauroneis and to form a new combination, Prestauroneis crucicula (W. Smith) Genkal et Yarushina comb. nov.     

The allantoplacenta of Mabuya mabouya (Sauria, Scincidae)

Analysis of placentation in the final stages of development in Mabuya mabouya shows that the placenta is formed by the apposition of the chorioallantois to the uterine mucosa implicating the entire embryonic chamber, because the allantoic vesicle invades all the exocoelom. The chorioallantoic placenta presents the features proper of a type IV allantoplacenta. However, in the mesometrial area peripheral to the placentome, we found that the paraplacentome is an additional zone specialized for histotrophic transfer, and is separated from the rest of the embryonic chamber by a chorionic invagination formed of polymorphic cells. The chorionic areolae are components of the embryonic hemisphere; they are in apposition to an endometrium with columnar epithelial cells and several glands that secrete toward the cavity of the areolae. They are observed only in the preparturition stage, probably operating in maternal-fetal transfer of nutrients during the last embryonic growth stage. The mesometrial hemisphere possesses specializations related to histotrophic nutrition (placentome, paraplacentome, and chorionic areolae), while in the abembryonic hemisphere there is an allantoplacenta of mixed function, with capacity for histotrophic nutrition and for gas exchange. The absorptive plaques are small rounded areas constituted by chorionic cells similar to the paraplacentomal chorionic cells, in intimate apposition with a secretory uterine epithelium. Separating the absorptive plaques are respiratory segments histologically similar to the type I allantoplacenta. The additional histotrophic areas found for this species demonstrate the great specialization of this allantoplacenta, and support the highest degree of matrotrophy among reptiles reached in the Neotropical Mabuya.     

De novo backbone trace of GroEL from single particle electron cryomicroscopy

In this work, we employ single-particle electron cryo-microscopy (cryo-EM) to reconstruct GroEL to approximately 4 A resolution with both D7 and C7 symmetry. Using a newly developed skeletonization algorithm and secondary structure element identification in combination with sequence-based secondary structure prediction, we demonstrate that it is possible to achieve a de novo Calpha trace directly from a cryo-EM reconstruction. The topology of our backbone trace is completely accurate, though subtle alterations illustrate significant differences from existing crystal structures. In the map with C7 symmetry, the seven monomers in each ring are identical; however, the subunits have a subtly different structure in each ring, particularly in the equatorial domain. These differences include an asymmetric salt bridge, density in the nucleotide-binding pocket of only one ring, and small shifts in alpha helix positions. This asymmetric conformation is different from previous asymmetric structures, including GroES-bound GroEL, and may represent a "primed state" in the chaperonin pathway.     

Large oligomeric complex structures can be computationally assembled by efficiently combining docked interfaces

Macromolecular oligomeric assemblies are involved in many biochemical processes of living organisms. The benefits of such assemblies in crowded cellular environments include increased reaction rates, efficient feedback regulation, cooperativity and protective functions. However, an atom‐level structural determination of large assemblies is challenging due to the size of the complex and the difference in binding affinities of the involved proteins. In this study, we propose a novel combinatorial greedy algorithm for assembling large oligomeric complexes from information on the approximate position of interaction interfaces of pairs of monomers in the complex. Prior information on complex symmetry is not required but rather the symmetry is inferred during assembly. We implement an efficient geometric score, the transformation match score, that bypasses the model ranking problems of state‐of‐the‐art scoring functions by scoring the similarity between the inferred dimers of the same monomer simultaneously with different binding partners in a (sub)complex with a set of pregenerated docking poses. We compiled a diverse benchmark set of 308 homo and heteromeric complexes containing 6 to 60 monomers. To explore the applicability of the method, we considered 48 sets of parameters and selected those three sets of parameters, for which the algorithm can correctly reconstruct the maximum number, namely 252 complexes (81.8%) in, at least one of the respective three runs. The crossvalidation coverage, that is, the mean fraction of correctly reconstructed benchmark complexes during crossvalidation, was 78.1%, which demonstrates the ability of the presented method to correctly reconstruct topology of a large variety of biological complexes. Proteins 2015; 83:1887–1899. © 2015 The Authors. Proteins: Structure, Function, and Bioinformatics Published by Wiley Periodicals, Inc.     

High incidence of supernumerary nipples and twins in formosan macaques (Macaca cyclopis) at Mt. Longevity, Taiwan

A population of Formosan macaques at Mt. Longevity exhibits an unusually high incidence of supernumerary nipples (polythelia: between 1-6 accessory nipples and/or areolae on 33% of adults), as well as a high rate of twinning (about 1% of births). The coexistence of these unusual traits suggests a connection, which is further supported by a tendency for mothers of twins to have accessory nipples and for twins to be born in troops with high incidence of polythelia.     

国产合欢属和棋子豆属的叶结构及其分类学意义

本文通过对国产合欢属Albizia 15种及棋子豆属Cylindrokelupha的叶结构观察,发现此两属叶结构特征有明显的区别,是较好的属间区别性状,表现在合欢属(3)4级以上脉至盲脉完全被厚壁的维管柱鞘细胞深深包埋,为包埋脉,且这些包埋脉宽度相等,不逐级递减,不易辨别,多分枝;网眼较小,发育不完善,不易辨别;而棋子豆属则表现为4级以上脉逐级递减,不为较厚的厚壁细胞的维管柱鞘细胞包埋,易于辨别,网眼接近中等大小,易辨。此外,还提供了国产合欢属15种,棋子豆属8种叶结构分析的基础资料,并就其在分类上的意义作了一些探讨。     

The biology of the crustose lichen <Emphasis Type="Italic">Rhizocarpon geographicum</Emphasis>

The crustose lichen Rhizocarpon geographicum (L.) DC. comprises yellow-green lichenized areolae which develop and grow on the surface of a non-lichenized fungal hypothallus, the latter extending beyond the edge of the areolae to form a marginal ring. The hypothallus advances very slowly and the considerable longevity of R. geographicum, especially in Arctic and Alpine environments, has been exploited by geologists in dating the exposure age of rock surfaces (lichenometry). This review explores various aspects of the biology of R. geographicum including: (1) structure and symbionts, (2) lichenization, (3) development of areolae, (4) radial growth rates (RaGR), (5) growth physiology, (6) changes in RaGR with thallus size (growth rate-size curve), (7) maturity and senescence, and (8) aspects of ecology. Lichenization occurs when fungal hyphae become associated with a compatible species of the alga Trebouxia, commonly found free-living on the substratum. Similarly, ‘primary’ areolae develop from free-living algal cells trapped by the advancing hypothallus. The shape of the growth rate-size curve of R. geographicum is controversial but may exhibit a phase of decreasing growth in larger thalli. Low rates of translocation of carbohydrate to the hypothallus together with allocation for stress resistance results in very slow RaGR, a low demand for nutrients, hence, the ability of R. geographicum to colonize more extreme environments. Several aspects of the biology of R. geographicum have implications for lichenometry including early development, mortality rates, the shape of the growth-rate size curve, and competition.     

Reconstruction of the burned ear using a temporalis fascial flap

A case report is presented in which a temporalis fascial flap is used to reconstruct a burned ear complicated by full-thickness skin loss with exposed cartilage. The method provides a simple, reliable means for covering exposed auricular cartilage with readily obtainable vascularized tissue. Donor-site morbidity is minimal because the scar lies within the hairline.