bluuebunny/crossref_metadata_2025 huggingface dataset:
Vegetationsökologische Untersuchungen auf nährstoffreichen Feuchtgrünland‐Brachen Sukzessionsverlauf und dynamisches Verhalten von Einzelarten
Authors: Jensen K.
Published: August 2010

Bis weit in unser Jahrhundert bestanden große Teile des Grünlandes des nordwestdeutschen Tieflandes aus extensiv bewirtschafteten artenreichen Feuchtwiesen. Eine Nutzungsaufgabe führt in diesen Systemen zu Verschiebungen im Konkurrenzgefüge der Pflanzenarten und löst Veränderungen in der Vegetationszusammensetzung und ‐struktur aus. Die Sukzession nach der Nutzungsaufgabe ist durch die Abfolge charakteristischer Brachestadien gekennzeichnet: Nach einer Initialphase (Brachestadium I) kommt es zur Ausbildung artenarmer Dominanzbestände hochwüchsiger Pflanzenarten, die bereits im Ausgangsbestand vorhanden waren (Brachestadium II). Der weitere Sukzessionsverlauf wird durch das Einwandern und die Etablierung von krautigen Arten ausgelöst, die in den Beständen zur Kodominanz gelangen (Brachestadium III). Schließlich kommt es zur Einwanderung und Etablierung von Gehölzen, so daß sich Gehölzbestände ausbilden (Brachestadium IV). Im Laufe der untersuchten Sukzession (Brachestadium I bis III) geht die Artenzahl ebenso zurück wie die Diversität und die Evenness. Es kommt zur Ausbildung hochproduktiver Bestände sowie zur Ausbildung einer mächtigen Streuschicht (bis zu 500 g/m2), so daß die Keimung und Etablierung vieler Arten behindert werden.

Die in der Sukzession auftretenden Arten können nach ihrem dynamischen Verhalten vier Sukzessionskategorien zugeordnet werden: Die Deckungsanteile der Arten der Sukzessionskategorie A und B gehen im Laufe der Sukzession zurück. Während Arten der Sukzessionskategorie A (z. B.Lychnis flos‐cuculi, Anthoxanthum odoratum) in späteren Sukzessionsstadien lokal aussterben, kommen Arten der Sukzessionskategorie B (z. B.Cirsium oleraceum. Ranunculus repens) in diesen regelmäßig in geringer Deckung vor. Die Deckungsanteile der Arten der Sukzessionskategorie C (z. B.Angelica sylvestris, Equisetum fluviatile) bleiben im Laufe der Sukzession weitgehend unverändert. Arten der Sukzessionskategorie D (Carex acutiformis, Galeopsis tetrahitagg.) erhöhen ihre Deckungsanteile in späten Brachestadien. Die Zuordnung der Arten zu den Sukzessionskategorien kann mit Wuchseigenschaften der Arten korreliert werden: Arten der Sukzessionskategorien A und B sind durch niedrige Wuchshöhen gekennzeichnet. Weiterhin sind viele der Arten auf eine regelmäßige Fortpflanzung durch Samen angewiesen. Arten der Sukzessionskategorie D verfügen über große Wuchshöhen, ihre Wuchsform ist oft durch unterirdische Ausläufer (Rhizome) gekennzeichnet. Die Arteigenschaften der Arten der Sukzessionskategorie C sind intermediär. einige Arten entgehen der Konkurrenz der hochwüchsigen Arten der Sukzessionskategorie D durch phänologische Differenzierung (z. B.Caltha palustris).

Abschließend werden die der Sukzession zugrunde liegenden Mechanismen und die Anwendbarkeit bestehender Sukzessionsmodelle auf die Vegetationsveränderungen auf Feuchtgrünland‐Brachen diskutiert.
Calvatia booniana(Lycoperdaceae) new from Europe and Asia
Authors: Moreno G. , Alté A. , Kreisel H.
Published: August 2010

Calvatia boonianaA. H. SM. (Lycoperdaceae) is recorded for the first time from Europe (Spain) and Asia (Iran and Nepal). Material from these countries is described and compared with the type collection from Oregon, U.S.A. That means the presence of a second species of “Giant Puffball” in Europe. The most important characters are illustrated by photographs with optic and SE microscope. Distribution and ecology of this and closely related species are compared.
Die GattungChamaesycein Europa. Bestimmungsschlüssel mit taxonomisch‐nomenklatorischen Anmerkungen
Authors: Hügin G.
Published: August 2010

Zwei Schlüssel ermöglichen die Bestimmung aller in Europa nachgewiesenen Arten der GattungChamaesyce(Euphorbiaceae); auch irrtümlich angegebene, nicht zweifelsfrei nachgewiesene und noch zu erwartende Taxa sind berücksichtigt.

Bestimmungsmerkmale sowie taxonomischnomenklatorische Probleme werden diskutiert.
Analysis of floristical and environmental gradients in the longitudinal profile of the Loire (France). The use of riparian plant species for a biotypological river‐zone classification
Authors: Wisskirchen R. , Kisteneich S. , Krause S.
Published: August 2010

The composition of riverbank vegetation at the French Loire was sampled in regular intervals along the longitudinal river profile. Distribution and abundance if 58 selected species of the willow and poplar woodlands, reed beds and adjacent therophyte communities are described from source to estuary.

The longitudinal profile of the Loire can be divided by key species groups into four floristically and ecologically defined sections. Their boundaries mainly fall together with changes of water regime characteristics.

The connections between species arrangement and major environmental factors (soil and water parameters) are investigated exemplarily for Therophyte communities colonizing riverbanks. The change of species composition (mathematically expressed as “floristic change index”) along the river shows a highly significant monotounous trend and is correlated with changes of pH and electrical conductivity of the soil.
Petiver's Gazophylacii and the identity ofDorotheanthus gramineus(HAW.) SCHWANTES (Aizoaceae)
Authors: Struck M.
Published: August 2010

A re‐examination of the holotype ofD. gramineus(HAW.) SCHWANTES in PETIVER's Gazophylacii established that this name has traditionally been misapplied. PETIVER's figure portrays a grassy annual with petals being shorter than the calyx lobes which clearly identify it as being conspecific with–and a younger synonym of –D. apetalus(L. f.) N. E. BR. The oldest name available to be accepted under the ICBN for the species hitherto misidentified as“D. gramineus”isMesembryanthemum clavatumHAW. which was likewise founded on a drawing in PETIVER's Gaz. The necessary new combinationD. clavatus(HAW.) STRUCK is published here.
Surface sculpturing ofAlstonia booneiiDE WILD. andA. congensisENGL. (Apocynaceae) and its importance on their taxonomy
Authors: Jayeola A. A.
Published: August 2010

SEM examination of the lower epidermis of mature leaves of the two species ofAlstoniaR. BR. (Apocynaceae) in Nigeria has provided constant characters potentially useful for their identification. The patterns of epicuticular sculpturing inA. booneiiDE WILD. andA. congensisENGL. differ in the size and density of knots, length of internodes, exposure or concealment of stomata, all knitted intricately to suggest their possible influence on wind speed, water loss and habit selection.
Ottleya, a new genus of Papilionaceae — Loteae from North America
Authors: Sokoloff D. D.
Published: August 2010

A new genus, Ottleya gen. nov., is described from North America. The genus Ottleya corresponds to Lotus group Simpeteria of ISELY's (1981) classification and comprises 12 species: O. argyraea comb, nov., O. flexuosa comb. nov., O. grandiflora comb. nov., O. greenei comb. nov., O. intricata comb. nov., O. mearnsii comb. nov., O. nivea comb. nov., O. oroboides comb. nov., O. rigida comb. nov., O. strigosa comb, nov., O. utahensis comb. nov. and O. wrightii comb. nov. restricted to SW part of the USA, N and Central Mexico.
The taxonomy of Fagus in western Eurasia. 2: Fagus sylvatica subsp. sylvatica
Authors: Denk Th.
Published: August 2010

The European beech (Fagus sylvatica subsp. sylvatica) is known to be tremendously variable both morphologically and genetically. By undertaking field studies and morphometrical analyses a number of recurrent morphological features turned out to cause this variability. These are mostly reactions of the genus to environmental conditions as to be found, e.g. in dry and warm sites or in humid gorges etc throughout the whole range of Fagus sylvatica subsp. sylvatica. Morphotypes in Italy, the former Yugoslavia, Hungary and Bulgaria, which were described by CZECZOTT (1933) as Fagus moesiaca, are well within the morphological range of Fagus sylvatica subsp. sylvatica and should be included in Fagus sylvatica s. str. Morphoclinal transitions can be observed in a WE direction, along a vertical belt and to a smaller extent in a NS direction. Only in the easternmost Bulgaria do hybrids with the Oriental beech (Fagus sylvatica subsp. orientalis) occur. Scattered populations of transitional forms, however, extend to North Greece, Central and SW Europe.

Basically, European beech populations are more homogeneous than those occurring from Turkey to Iran. This might be explained by fewer and smaller refugia during the Pleistocene period as well as by the late initiation of postglacial recolonization in Europe.
A synopsis of the genusAtriplexL. (Chenopodiaceae) in Saudi Arabia
Authors: Al‐Turki T. A. , Omer S. , Ghafoor A.
Published: August 2010

The genusAtriplexL. is revised from Saudi Arabia. In all 10 species are reported, out of which seven are native(Atriplex coriacea, A. dimorphostegia, A. farinosa, A glauca, A. halimus, A. leucocladaandA. tatarica), and three are introduced and cultivated/naturalised(A. canescens, A. semibaccataandA. suberecta). The study has introduced the presences of following new taxa:Atriplex dimorphostegiaf.collenetteaAl‐Turki, Omer & A. Ghafoor f. nova, andAtriplex leucocladavar.mandavilleaAl‐Turki, Omer & A.Ghafoor var. nova. Status and history of genera segregated fromAtriplexhas been discussed. A key for identification of taxa recognized and their descriptions are given; notes on nomenclature, variation pattern, ecology, distribution maps, local names and uses are provided. A list of introduced and cultivated species is appended.
A combined top‐down and bottom‐up MS approach for the characterization of hemoglobin variants in Rhesus monkeys
Authors: Hüttenhain Ruth , Hess Sonja
Published: August 2010

Sickle cell disease is caused by one of the 1200 known hemoglobin variations. A single‐point mutation β6(A3)Glu→Val leads to sickling of red blood cells, which in turn causes a lack of oxygen supply to tissue and organs. Although sickle cell disease is well understood, treatment options are currently underdeveloped. The only Food and Drug Administration‐approved drug is hydroxyurea, an inducer of fetal γ‐hemoglobin, which is known to have a higher oxygen affinity than adult hemoglobins and thus alleviates symptoms. In the search for better cures, Rhesus monkeys (Macaca mulatta) serve as models for monitoring success of induction of fetal γ‐hemoglobins and with recent advances in proteomics, MS has become the leading technique to determine globin expression. Similar to humans, Rhesus monkeys possess hemoglobin variants that have not been sufficiently characterized to initiate such a study. Therefore, we developed a combined bottom‐up and top‐down approach to identify and characterize novel hemoglobin variants of the umbilical cord blood of Rhesus monkeys. A total of four different variants were studied: α, β, γ1 and γ2. A new α‐ and β‐hemoglobin variant was identified, and the two previously hypothesized γ‐hemoglobins were identified. In addition, glutathionylation of both γ‐hemoglobin variants at their cysteines has been characterized. The combined approach outperformed either bottom‐up or top‐down alone and can be used for characterization of unknown hemoglobin variants and their PTMs.
Poor outcome following aborted orthotopic liver transplantation due to severe porto‐pulmonary hypertension
Authors: Fukazawa Kyota , Pretto Ernesto A.
Published: August 2010

Background/purpose

The diagnosis of porto‐pulmonary hypertension (PPHN) in patients with end‐stage liver disease at the time of surgery is not an uncommon occurrence. The decision to proceed with orthotopic liver transplantation (OLT) in patients with severe PPHN is associated with high perioperative mortality. We sought to determine the progression of PPHN and patient outcome following aborted OLT.

Methods

A retrospective analysis of 2150 OLT cases was performed.

Results

The analysis revealed that six cases (0.3%) were cancelled due to severe PPHN at the time of surgery. The progression to severe PPHN occurred on median over a period of 82 days (range 10–229 days). Following aborted OLT, three patients (50%) expired shortly after.

Conclusions

Cancellation of OLT due to severe pulmonary hypertension was also associated with high mortality. Moreover, this preliminary study reveals that once diagnosed, PPHN can progress to a more severe form over a short period of time. Therefore, we recommend more frequent monitoring of PPHN in this population of patients.
The distance of tumor spread in the main pancreatic duct of an intraductal papillary‐mucinous neoplasm: where to resect and how to predict it
Authors: Okada Ken‐ichi , Imaizumi Toshihide , Hirabayashi Kenichi , Matsuyama Masahiro , Yazawa Naoki , Dowaki Shoichi , Tobita Kosuke , Ohtani Yasuo , Kawaguchi Yoshiaki , Tanaka Makiko , Inokuchi Sadaki , Makuuchi Hiroyasu
Published: August 2010

Background

The surgical decision regarding where to resect the pancreas is an important judgement that is directly linked to the surgical procedure. An appropriate surgical margin to resect intraductal papillary‐mucinous neoplasm (IPMN) of the pancreas based on the distance of tumor spread (DTS) in the main pancreatic duct has not been adequately documented. We analyzed the appropriate surgical margin based on the DTS in the main pancreatic duct of IPMN and the positive rate at the pancreatic cut end margin.

Methods

Forty patients with main duct‐ or mixed‐type IPMN diagnosed histopathologically who underwent surgery at Tokai University Hospital between 1991 and 2008 were retrospectively analyzed. The resection line was determined to achieve a 2‐cm surgical margin in patients with main duct‐ or mixed‐type IPMN and as limited a resection as possible to remove the dilated branch duct in patients with branch duct‐type IPMN according to macroscopic type. The dysplastic state of the epithelium was judged as positive for carcinoma in situ (high‐grade dysplasia) or adenoma (very low to moderate dysplasia) and judged as negative for hyperplasia or normal.

Results

The mean DTS in the main pancreatic duct was 41.6 ± 30.0 mm, and that of the distance of tumor absence was 13.6 ± 12.4 mm. The positive rate at the pancreatic cut end margin in frozen sections was 29.7%. The final positive rate at the pancreatic cut end margin was 26.2%. There has been no evidence of local recurrence in the remnant pancreas. DTS in the main pancreatic duct of IPMN was correlated with the maximum diameter of the duct (R = 0.678).

Conclusion

Distance of tumor spread offered important insights about the appropriate site to resect the pancreas and the positive rate at the cut end margin in IPMN.
Organic–Inorganic Coatings Based on Epoxidised Castor Oil/APTES/TEOS
Authors: Becchi Denize Maria , de Luca Maria Augusta , Martinelli Márcia , Mitidieri Sydnei
Published: August 2010

Two series of organic–inorganic hybrid films were prepared from epoxidised castor oil (ECO) and the inorganic precursor 3‐aminopropyltriethoxysilane (APTES), and the combination of APTES with tetraethoxysilane (TEOS) with different organic to inorganic proportions. Films were pre‐cured at room temperature under inert atmosphere and subsequently submitted to thermal curing. The macro‐ and microscopic properties of the films, including adhesion, hardness, microstructure and thermal properties, were determined as a function of the proportion of ECO to inorganic precursors. Morphologic studies showed that the hybrid films were microscopically homogeneous. The hardness and tensile strength of the films increased with increased concentrations of inorganic precursor. All of the films exhibited good adhesion to an aluminium surface and worked as an efficient barrier against corrosion.
Analysis of Lard's Aroma by an Electronic Nose for Rapid Halal Authentication
Authors: Nurjuliana M. , Che Man Y. B. , Mat Hashim D.
Published: August 2010

An electronic nose was successfully used to detect and discriminate lard from other types of animal body fats and samples containing lard. The results are presented in the form of VaporPrint™, the image of the polar plot of the odor amplitudes from the surface acoustic wave (SAW) detector frequency. In the VaporPrint™, the radial angles representing the sensor provides individual fingerprints of the aroma of different animal body fats. Principal component analysis (PCA) was used to interpret the data and it provided a good grouping of samples, with 61% of the variation accounted for by PC 1 and 29% accounted for by PC 2. All of the lard‐containing samples formed a separate group from the samples that were free from lard. This method can be developed into a rapid method for detecting the presence of lard in food samples for Halal authentication.
Two novel mutations of SCN9A (Nav1.7) are associated with partial congenital insensitivity to pain
Authors: Staudl Roland , Pricel Donald D. , Janickel David , Andradel Edgard , Hadjipanayisl Angela G. , Eatonl Will T. , Kaplanl Lee , Wallacel Margaret R.
Published: August 2010

Insensitivity to pain is a rare disorder that is commonly associated with Hereditary Sensory and Autonomic Neuropathies (HSAN I—V) resulting often in autonomic dysfunction and premature death. Very few individuals have been reported with pain insensitivity lacking such autonomic neuropathies. We performed genetic, neurologic, psychological, and psychophysical evaluations in such an individual (OMIM 243000) and her first degree relatives. Sequence analysis of genomic DNA revealed two novel SCN9A mutations in this index case (IC). One was a non‐conservative missense mutation (C1719R) in exon 26 present only in the IC and one parent. Further sequence analysis of the chilD's DNA revealed a 1‐bp splice donor deletion in intron 17 which was also present in the other parent and one sibling. Detailed psychophysical testing was used to phenotypically characterize the IC, her family members, and 10 matched normal controls. Similar to family members and controls the IC showed normal somatosensory functioning for non‐nociceptive mechanoreception and warmth. However, she demonstrated diminished ability to detect cool temperatures combined with profound deficits in heat and mechanical nociception. Congenital insensitivity to pain in our IC was associated with two novel SCN9A mutations which most likely resulted in a Nav1.7 channelopathy. However, in contrast to individuals with other SCN9A mutations, the observed pain insensitivity was relative and not absolute, which may be consistent with hypomorphic effects of one or both mutations. The ability to sense at least some danger signals may be advantageous and ameliorate the otherwise increased morbidity and mortality of some individuals with congenital insensitivity to pain.
POSH2 is a RING finger E3 ligase with Rac1 binding activity through a partial CRIB domain
Authors: Kärkkäinen Satu , van der Linden Maarten , Renkema G. Herma
Published: August 2010

MINT‐7987920: PAK2 (uniprotkb:Q13177) binds (MI:0407) to Rac1 (uniprotkb:P63000) by solid phase assay (MI:0892)
Tankyrase‐1 assembly to large protein complexes blocks its telomeric function
Authors: Hatsugai Kaori , Ohishi Tomokazu , Sugimoto Yoshikazu , Seimiya Hiroyuki
Published: August 2010

MINT‐7987998: Tankyrase‐1 (uniprotkb:O95271) physically interacts (MI:0915) with Tankyrase‐1 (uniprotkb:O95271) by anti tag coimmunoprecipitation (MI:0007).
Structural identity of telomeric complexes
Authors: Giraud-Panis Marie-Josèphe , Pisano Sabrina , Poulet Anaïs , Du Marie-Hélène Le , Gilson Eric
Published: August 2010

A major issue in telomere research is to understand how the integrity of chromosome ends is controlled. Although several nucleoprotein complexes have been described at the telomeres of different organisms, it is still unclear how they confer a structural identity to chromosome ends in order to mask them from DNA repair and to ensure their proper replication. In this review, we describe how telomeric nucleoprotein complexes are structured, comparing different organisms and trying to link these structures to telomere biology. It emerges that telomeres are formed by a complex and specific network of interactions between DNA, RNA and proteins. The fact that these interactions and associated activities are reinforcing each other might help to guaranty the robustness of telomeric functions across the cell cycle and in the event of cellular perturbations. We propose that telomeric nucleoprotein complexes orient cell fate through dynamic transitions in their structures and their organization.
Mast cell signaling: The role of protein tyrosine kinase Syk, its activation and screening methods for new pathway participants
Authors: Siraganian Reuben P. , de Castro Rodrigo O. , Barbu Emilia A. , Zhang Juan
Published: August 2010

The aggregation by antigen of the IgE bound to its high affinity receptor on mast cells initiates a complex series of biochemical events that result in the release of inflammatory mediators. The essential role of the protein tyrosine kinase Syk has been appreciated for some time, and newer results have defined the mechanism of its activation. The use of siRNA has defined the relative contribution of Syk, Fyn and Gab2 to signaling and has made possible a screening study to identify previously unrecognized molecules that are involved in these pathways.
Vegetation development in the Middle Euphrates and Upper Jazirah (Syria/Turkey) during the Bronze Age
Authors: Deckers Katleen , Pessin Hugues
Published: August 2010

Vegetation changes are reconstructed based on more than 51,000 charcoal fragments of more than 380 samples from nine Bronze Age sites in northern Syria and southern Turkey. In addition to fragment proportions, special attention was paid to the frequency of Pistacia relative to Quercus and Populus/Salix relative to Tamarix, fruit-tree ubiquity, and riverine diversity in order to gain an improved understanding of the human versus climatic impact on the vegetation. The results indicate that human impacts first took place within the riverine forest. This phase was followed by land clearing within the woodland steppe, especially in the northern portion of the study area. In the south near Emar, the woodland steppe probably disappeared by the Late Bronze Age. It is uncertain whether this was caused by aridification and/or human clearing. The northward shift of the Pistacia-woodland steppe is very likely a result of climatic drying that occurred throughout the entire period under investigation. Although increased deforestation is evident through time, the small proportions of imported wood indicate that local resources were still available.
Testing the Landscape Reconstruction Algorithm for spatially explicit reconstruction of vegetation in northern Michigan and Wisconsin
Authors: Sugita Shinya , Parshall Tim , Calcote Randy , Walker Karen
Published: August 2010

The Landscape Reconstruction Algorithm (LRA) overcomes some of the fundamental problems in pollen analysis for quantitative reconstruction of vegetation. LRA first uses the REVEALS model to estimate regional vegetation using pollen data from large sites and then the LOVE model to estimate vegetation composition within the relevant source area of pollen (RSAP) at small sites by subtracting the background pollen estimated from the regional vegetation composition. This study tests LRA using training data from forest hollows in northern Michigan (35 sites) and northwestern Wisconsin (43 sites). In northern Michigan, surface pollen from 152-ha and 332-ha lakes is used for REVEALS. Because of the lack of pollen data from large lakes in northwestern Wisconsin, we use pollen from 21 hollows randomly selected from the 43 sites for REVEALS. RSAP indirectly estimated by LRA is comparable to the expected value in each region. A regression analysis and permutation test validate that the LRA-based vegetation reconstruction is significantly more accurate than pollen percentages alone in both regions. Even though the site selection in northwestern Wisconsin is not ideal, the results are robust. The LRA is a significant step forward in quantitative reconstruction of vegetation.
An indirect assessment on the impact of connectivity of conductivity classes upon longitudinal asymptotic macrodispersivity
Authors: Fiori Aldo , Boso Francesca , de Barros Felipe P. J. , De Bartolo Samuele , Frampton Andrew , Severino Gerardo , Suweis Samir , Dagan Gedeon
Published: August 2010

Solute transport takes place in heterogeneous porous formations, with the log conductivity, Y = ln K, modeled as a stationary random space function of given univariate normal probability density function (pdf) with mean 〈Y〉, variance σY2, and integral scale IY. For weak heterogeneity, the above mentioned quantities completely define the first‐order approximation of the longitudinal macrodispersivity αL = σY2IY. However, in highly heterogeneous formations, nonlinear effects which depend on the multipoint joint pdf of Y, impact αL. Most of the past numerical simulations assumed a multivariate normal distribution (MVN) of Y values. The main aim of this study is to investigate the impact of deviations from the MVN structure upon αL. This is achieved by using the concept of spatial correlations of different Y classes, the latter being defined as the space domain where Y falls in the generic interval [Y,Y + ΔY]. The latter is characterized by a length scale λ(Y), reflecting the degree of connectivity of the domain (the concept is similar to the indicator variograms). We consider both “symmetrical” and “non‐symmetrical” structures, for which λ(Y′) = λ(−Y′) (similar to the MVN), and λ(Y′) ≠ λ(−Y′), respectively, where Y′ = Y − 〈Y〉. For example, large Y zones may have high spatial correlation, while low Y zones are poorly correlated, or vice versa. The impact of λ(Y) on αL is investigated by adopting a structure model which has been used in the past in order to investigate flow and transport in highly heterogeneous media. It is found that the increased correlation in the low conductive zones with respect to the high ones generally leads to a significant increase in αL, for the same global IY. The finding is explained by the solute retention occurring in low Y zones, which has a larger effect on solute spreading than high Y zones. Conversely, αL decreases when the high conductivity zones are more correlated than the low Y ones. Dispersivity is less affected by the shape of λ(Y) for symmetrical distributions. It is found that the range of validity of the first‐order dispersivity, i.e., αL = IYσY2, narrows down for non‐symmetrical structures.
The inhibition of monocarboxylate transporter 2 (MCT2) by AR-C155858 is modulated by the associated ancillary protein
Authors: Ovens Matthew J. , Manoharan Christine , Wilson Marieangela C. , Murray Clarey M. , Halestrap Andrew P.
Published: August 2010

In mammalian cells, MCTs (monocarboxylate transporters) require association with an ancillary protein to enable plasma membrane expression of the active transporter. Basigin is the preferred binding partner for MCT1, MCT3 and MCT4, and embigin for MCT2. In rat and rabbit erythrocytes, MCT1 is associated with embigin and basigin respectively, but its sensitivity to inhibition by AR-C155858 was found to be identical. Using RT (reverse transcription)–PCR, we have shown that Xenopus laevis oocytes contain endogenous basigin, but not embigin. Co-expression of exogenous embigin was without effect on either the expression of MCT1 or its inhibition by AR-C155858. In contrast, expression of active MCT2 at the plasma membrane of oocytes was significantly enhanced by co-expression of exogenous embigin. This additional transport activity was insensitive to inhibition by AR-C155858 unlike that by MCT2 expressed with endogenous basigin that was potently inhibited by AR-C155858. Chimaeras and C-terminal truncations of MCT1 and MCT2 were also expressed in oocytes in the presence and absence of exogenous embigin. L-Lactate Km values for these constructs were determined and revealed that the TM (transmembrane) domains of an MCT, most probably TM7–TM12, but not the C-terminus, are the major determinants of L-lactate affinity, whereas the associated ancillary protein has little or no effect. Inhibitor titrations of lactate transport by these constructs indicated that embigin modulates MCT2 sensitivity to AR-C155858 through interactions with both the intracellular C-terminus and TMs 3 and 6 of MCT2. The C-terminus of MCT2 was found to be essential for its expression with endogenous basigin.
PEGylated Gold Nanoparticles Functionalized with β-Cyclodextrin Inclusion Complexes: Towards Metal Nanoparticle - Polymer - Carbohydrate Cluster Biohybrid Materials
Authors: Liu Jingquan , Setijadi Eki , Liu Yingkai , Whittaker Michael R. , Boyer Cyrille , Davis Thomas P.
Published: August 2010

A cholesterol-functional trithiocarbonate reversible addition–fragmentation chain transfer (RAFT) agent was synthesized and employed to generate well-defined poly(polyethylene glycol) acrylate with cholesterol chain termini using RAFT polymerization. Subsequently, the polymers were grafted onto the surface of gold nanoparticles using the trithiocarbonate functionality to bind to the gold surface. The cholesterol moieties were then modified via complexation with β-cyclodextrin. The step-by-step modification of gold nanoparticles was characterized by dynamic light scattering, attenuated total reflection infrared spectroscopy and surface plasmon resonance analysis.
Bovine embryo transfer recipient synchronisation and management in tropical environments
Authors: Baruselli Pietro S. , Ferreira Roberta M. , Filho Manoel F. Sá , Nasser Luiz F. T. , Rodrigues Carlos A. , Bó Gabriel A.
Published: August 2010

Numerous studies have shown that it is possible to manipulate follicular and luteal dynamics, thereby eliminating the need for oestrus detection in embryo transfer (ET) programmes. Fixed-time ET (FTET) protocols are based on the use of gonadotrophin-releasing hormone (GnRH) and prostaglandin (PG) F or progesterone/progestogen (P4)-releasing devices and oestradiol. The FTET protocols increases the proportion of recipients transferred, and therefore pregnancy rates, compared with the use of PGF followed by ET 7 days after oestrus. Furthermore, the addition of equine chorionic gonadotrophin (eCG) to the P4 and oestradiol-based FTET protocols results in an even higher proportion of recipients transferred, and thus higher pregnancy rates. The beneficial effect of eCG treatment may be related to increased growth of the dominant follicle and increased plasma P4 concentrations during the subsequent luteal phase. In Bos taurus × Bos indicus recipients, pregnancy rates were positively correlated with the diameter of the corpus luteum (CL) and the number of CL at ET. When repeat-breeder Holstein cows were used as recipients, FTET protocols increased number of recipients transferred and pregnancy rates compared with the traditional PGF-based synchronisation protocols. In conclusion, the use of FTET protocols eliminates the need for the detection of oestrus and results in a greater proportion of recipients transferred and satisfactory pregnancy rates. Thus, FTET optimises the use of recipients, reducing labour and animal handling and facilitating the use of ET.
Practical applications of new research information in the practice of bovine embryo transfer
Authors: Looney C. R. , Pryor J. H.
Published: August 2010

For more than 40 years, practitioners have sought to improve all aspects of commercial bovine embryo transfer. The development of new technologies for this industry has been substantial, with recent focus on cryopreservation techniques and the in vitro production of embryos fertilised with sexed spermatozoa. When these and other new technologies are developed, the following questions remain: (1) is said technology regulated or does it require licensing; and (2) is it applicable and, if so, is it financially feasible? Computer access to published research and the advancement of data software programs conducive to the industry for data procurement have been essential for helping practitioners answer these questions by enhancing their ability to analyse and apply data. The focus of the present paper is to aid commercial embryo transfer practitioners in determining new technologies that are available and whether they can be implemented effectively, benefiting their programs.
Is the zona pellucida an efficient barrier to viral infection?
Authors: Van Soom A. , Wrathall A. E. , Herrler A. , Nauwynck H. J.
Published: August 2010

Although the transfer of embryos is much less likely to result in disease transmission than the transport of live animals, the sanitary risks associated with embryo transfer continue to be the subject of both scientific investigations and adaptations of national and international legislation. Therefore, the implications are important for veterinary practitioners and livestock breeders. In vivo-derived and in vitro-produced embryos are widely used in cattle and embryos from other species, such as sheep, goats, pigs and horses, are also currently being transferred in fairly significant numbers. Bearing in mind the wide variety of embryos of different species and the correspondingly large number of viruses that are of concern, it is expedient at this time to look again at the importance of the zona pellucida (ZP) as a barrier against viruses and at the susceptibility or otherwise of embryonic cells to viral infection if ever they are exposed. For embryos with an intact ZP, viral infection of the embryo is unlikely to occur. However, the virus may stick to the ZP and, in this case, International Embryo Transfer Society (IETS) washing procedures in combination with trypsin treatment are mandatory. A caveat is the fact that currently more and more types of embryos are becoming available for transfer and scientific data cannot be extrapolated from one species to another. These topics are discussed in the present review.
117 EFFECT OF DIFFERENT CRYOPRESERVATION METHODS ON THE QUALITY OF IN VITRO-PRODUCED BOVINE EMBRYOS
Authors: Stinshoff H. , Brüning K. , Hanstedt A. , Müller D. , Wilkening S. , Wrenzycki C.
Published: August 2010

In vitro production (IVP) of bovine embryos has been greatly improved over the last couple of years. However, only one-third of the total number of embryos transferred worldwide are of in vitro origin. The IVP embryos still show remarkable differences compared with their in vivo-derived counterparts (i.e. bovine embryos produced in vitro are more sensitive to cryopreservation). So far, vitrification seems to be the most promising method to cryopreserve in vitro-produced bovine embryos. The aim of this study was to determine the effect of 2 different cryopreservation methods on the quality of in vitro-produced bovine embryos at the molecular level using a sensitive RT-qPCR assay. Bovine blastocysts were produced using abattoir ovaries and a standard protocol for IVP (Wrenzycki et al. 2001). They were randomly vitrified employing PBS plus ethylene glycol and DMSO or cryopreserved using a programmable freezer and 1.5 M ethylene glycol. After thawing, embryos from both groups were cultured for 48 h. After 24 h of culture re-expansion rates were documented, and after 48 h hatching rates were documented. After hatching, blastocysts were stored at -80°C for subsequent RT-qPCR analysis. The following gene transcripts known to play important roles during preimplantation development were analyzed: HSP70, GLUT-1, GLUT-3, E-CAD, ZO-1, DNMT3a, IFNτ, DCII. Re-expansion rates were 74.7% (68/91) and 75.0% (87/116) for vitrified and conventionally cryopreserved blastocysts, and 57.1% (52/91) and 55.2% (64/116) of re-expanded embryos hatched. The relative abundances of HSP70, GLUT-1, and ZO-1 transcripts were significantly affected in both groups of cryopreservation compared with the control group (hatched blastocysts without cryopreservation). Conventional cryopreservation had a significant effect on the amount of GLUT-3, DNMT3a, and IFNτ mRNA, whereas vitrification significantly affected DCII transcripts. E-CAD mRNA expression was similar in all groups of embryos. These results suggest that not only the cryopreservation process itself but also the method used to freeze the embryos had a significant influence on the mRNA expression of developmentally important genes in hatched bovine blastocysts.

The support of the H.W. Schaumann Stiftung (Germany) and Gynemed Medizinprodukte GmbH & Co. KG (Germany) is gratefully acknowledged.
133 MITOCHONDRIAL ACTIVITY OF IN VIVO DEVELOPING PIG EMBRYOS
Authors: Romek M. , Gajda B. , Rolka M. , Smorag Z.
Published: August 2010

Preimplantation pig embryos undergo modifications in their metabolism. Glucose is metabolized via aerobic and anaerobic pathways at early cleavage stages, whereas glycolysis becomes the dominant pathway as development progresses. Moreover, fatty acid oxidation rate increases significantly at the early blastocyst stage. It is possible that mitochondria undergo structural and functional changes in response to altering embryo metabolism. Therefore, the aim of this study was to estimate changes in surface density of inner mitochondrial membranes (Sv) and mitochondrial activity (Av) per unit volume of mitochondria during the maturation of oocyte and early development of porcine embryos derived in vivo. The measurements were carried out on immature and matured oocytes, zygotes, 4 to 8 cell embryos, morula, blastocysts, and late and hatched blastocysts (10 to 39 embryos per stage). To estimate Sv, material was stained with mixture of 1.05 μM 10-N-nonyl Acridine Orange (NOA) and 0.5 μM Mito Tracker Deep Red (MtDR) for 30 min at 20°C. To measure Av, oocytes and embryos were labeled sequentially with 0.5μM Mito Tracker Orange CMTMRos (MtOR) for 30 min at 39°C and 0.5 μM MtDr for 30 min at 20°C. Embryos were then fixed in 3.8% formaldehyde and analyzed by confocal microscopy with a LSM 510 Meta Zeiss. The amounts of fluorescence emitted from embryos labeled as INOA, IMtOR, and IMtDR were measured because these ratios were directly proportional to Sv and Av, respectively. Values Sv and Av calculated for embryos at different stages of development were compared by ANOVA and Tukey’s intervals. During oocyte maturation, Av was low and remained unchanged, whereas Sv significantly decreased (P < 0.05) from 0.26 ± 0.021 f.a.u. (fluorescence arbitrary units) at immature oocytes to 0.11 ± 0.012 f.a.u. at matured oocytes. From the zygote to morula stage, Av was constant and then increased almost 5 times to the value of 0.95 ± 0.036 f.a.u. at the blastocyst stage, whereas late and hatched blastocyst characterized by significantly lower (P < 0.001) Av: 0.51 ± 0.023 and 0.34 ± 0.019 f.a.u., respectively. Surface density of the inner mitochondrial membrane in pig embryo during cleavage significantly (P < 0.05) increased from 0.16 ± 0.013 f.a.u. at zygote to 0.35 ± 0.020 and 0.75 ± 0.059 f.a.u. at morula and blastocyst, respectively. In conclusion, during the early stage of pig embryo development, mitochondria undergo structural and functional changes. Results demonstrate that the area density of inner mitochondrial membranes increases earlier (at 4 to 8 cell stages) than mitochondrial activity, which increase not before blastocyst stage. Furthermore, after blastocoel formation, when energy requirement is significantly lower, Av decrease but Sv remain constant.
149 EFFECTS OF FSH ADDED IN DEFINED MEDIUM MATURATION ON THE PRE-IMPLANTATION DEVELOPMENT AND EXPRESSION OF Bax, Bcl-2, AND CONNEXIN 43 GENES IN BOVINE IVP BLASTOCYSTS
Authors: Gulart L. V. M. , Gabriel L. , Salles L. P. , Gamas G. R. , Souza D. K. , Rosa e Silva A. A. M.
Published: August 2010

FSH at low concentrations affect embryo production. In vitro culture conditions also affect embryo production and embryonic expression of genes and alter oocyte competence to produce embryos. The search for better and less variable culture conditions simulating those in vivo has led to the development of several systems of oocyte in vitro maturation culture. To compare the efficiency of the systems of MIV we utilized 4 groups: (1) TCM-199 control; (2) α-minimal essential medium (MEM); 3) α-MEM + 1 ng of FSH; 4) α-MEM+ 10 ng of FSH. The medium of Group 1 is non-defined by the presence of fetal calf serum (10%). Groups 2, 3, and 4 are defined and polyvinyl alcohol (1%) was used as a macromolecule. Porcine FSH (1 IU mg-1) was used at 1 and 10 ng mL-1 and at 100 ng in defined and non-defined medium, respectively. Bovine ovaries were collected at an abbatoir. Oocytes (n = 1718) with homogeneous cytoplasm and with more than 3 layers of granulosa cells were used. Mature oocytes from the 4 treatments (11 replicates of each treatment) were inseminated with frozen-thawed, motile sperm separated by Percoll, using Sperm TALP HEPES medium. Presumptive zygotes with up to 2 or 3 layers of cumulus cells were cultured in 50-mL drops of SOF medium, supplemented with 10% FCS and 1 mg mL-1 BSA under mineral oil in a humid 5% CO2 atmosphere at 38.5°C after. Cleavage rate was evaluated 72 h post-insemination (hpi), and blastocyst rate was evaluated 168-192 hpi. Cleavage and blastocyst rates were calculated on the basis of number of presumptive zygotes. The expression of the following genes (Bax, Bcl-2, and conexin 43) was evaluated in blastocysts by RT-PCR. One-way ANOVA was used to compare blastocyst number. There was no difference in the proportion of embryos with more than 8 blastomeres in all groups tested, indicating that the rate of development during the first 72 hpi was similar for oocytes matured in chemically defined medium and for oocytes matured in medium containing serum. Bax is a pro-apoptotic marker and Bcl-2 an antiapoptotic marker. Connexin 43 (Cx43) may be a marker of embryo competence. Glyceraldehyde 3-phosphate dehydrogenase was used as internal control. The Bax gene was not expressed in any group. The Bcl-2 and Cx43 genes were expressed, mainly in the α-MEM 10. Although no differences were observed in blastocyst rate among the groups (30% to 40%), the strong expression of Bcl-2 and of Cx43 on the group containing 10 ng mL-1 of FSH may indicate that FSH could improve embryo quality under defined conditions.

The authors thank FAP-DF, CNPq, FUNPE, FINATEC, CAPES, and Biovitro Tecnologia de Embrioes Ltda, for laboratory assistance and grants, and Frigorifico Ponte Alta, Brasília-DF, for supplying bovine ovaries.
151 COMPARISON OF CELL NUMBERS OF ZONA-INTACT AND ZONA-FREE PARTHENOGENETICALLY ACTIVATED PORCINE EMBRYOS CULTURED IN VITRO
Authors: Li J. , Vjata G. , Callesen H.
Published: August 2010

Application of an artificial stimulus to activate oocytes and induce development is essential for the successful animal cloning by nuclear transfer (Zhu et al. 2002 Biol. Reprod. 66, 635-641). The embryo’s developmental competence could be further improved with optimal in vitro culture conditions (Du et al. 2007 Theriogenology 68, 1104-1110). Cell number determination is a commonly used and simple criterion to assess developmental quality of pre-implantation stage mammalian embryos (Lagutina et al. 2007 Theriogenology 67, 90-98). Our aim of the study was to investigate porcine embryos activated and cultured in different ways using total cell numbers as the only quality measure. After 43-44 h of in vitro maturation and cumulus cell removal, zona-intact (PAZI) or zona-free oocytes (PAZF; after pronase treatment) were subjected to parthenogenetic activation (Day 0) with a single 80-μs DC pulse of 1.26 kV cm-1 or 0.86 kV cm-1 (Kragh et al. 2005 Theriogenology 64, 1536-1545), followed by a 4-h treatment with 5 μg mL-1 of cytochalasin B and 10 μg mL-1 of cycloheximide. Subsequently, the well of the well system (Vajta in vitro 2000 Mol. Reprod. Dev. 55, 256-264) was used for culture of all PAZF and half of the PAZI embryos (PAZF-WOW and PAZI-WOW groups, respectively), whereas the remaining PAZI embryos were cultured in groups of 25-30 (PAZI group). All cultures were performed in porcine zygote medium 3 (Yoshioka et al. 2002 Biol. Reprod. 66, 112-119). On Day 6, all these in vitro cultured embryos were fixed and stained with Hoechst 33342 and cell numbers were counted on the microscopic pictures taken using fluorescent light. Data analysis was performed using ANOVA. Four replicates were performed with a total of 462 PAZF-WOW, 484 PAZI-WOW, and 467 PAZI group embryos. Embryos of each group were then divided into 5 groups based on their cell number (<5 cells, 5-8 cells, 9-16 cells, 17-32 cells, >32 cells). Percentages of embryos in each group are shown in Table 1. The average cell numbers of zona-intact embryos from PAZI-WOW and the PAZI group were similar to each other (P > 0.05), whereas the cell numbers of PAZF-WOW embryos were significantly different from both PAZI-WOW and PAZI group embryos (P < 0.05), with more embryos having higher cell numbers.The results demonstrate that zona-free parthenogenetically activated embryos cultured in WOW have higher cell numbers than embryos with intact zona pellucida. Accordingly, the presence of zona pellucida may compromise embryo development under certain in vitro culture situations.

Table 1.Distribution (in %) of parthenogenetically activated porcine embryos according to their cell number on Day 6 after activation
169 EMBRYO TRANSFER OF SEXED/VITRIFIED IVF EMBRYOS IN CATTLE: PREGNANCY COMPARISON AFTER SINGLE AND DOUBLE TRANSFERS
Authors: Castro A. S. , Xu J. , Pereira D. C. , Ferre L. , Diaz N. , Moreno J. , Du F.
Published: August 2010

Advancement in sperm sorting technology combined with vitrification of in vitro produced bovine embryos will promote cattle breeding and production. The objective of this study was to evaluate pregnancy and embryo loss after embryo transfer (ET) of sexed/vitrified embryos with one bilaterally (double transfer, 2 embryos) v. ipsilaterally (single transfer, 1 embryo) into the recipient. Bovine oocytes collected from slaughterhouse ovaries or ovum pickup were matured for 20-22 h, then subjected to IVF using Brackett and Oliphant BO procedures with sorted X-sperm, and cultured with our standard culture system. Expanded blastocysts with tight compaction of the inner cell mass (quality 1) were selected on Day 7 for cryopreservation via liquid nitrogen surface vitrification (LNSV; Xu et al. 2006 J. Dairy Sci. 89, 2510-2518). Embryo transfer was performed for 3 replicates in Navasota, Texas, in April 2009. Prior to ET, embryos were warmed and subsequently washed several times in warming, dehydration solution and base medium. Some of sexed/vitrified embryos were cultured for 3 days post-warming to determine the survivability. The treatments were as follows: (1) vitrified-single transfers, 1 embryo was transferred into the horn ipsilateral to CL; (2) vitrified-double transfers, 1 embryo was transferred into each uterine horn by nonsurgical transfer; and (3) fresh-single, 1 fresh embryo was transferred into the horn ipsilateral to CL (control) to a synchronous recipient on Day 7. Pregnancy was determined by ultrasound monitoring on Day 35, and palpation per rectum on Day 75 after transfer. The pregnancy data were analyzed by General Linear Model analysis (SPSS 11.0, SPSS Inc., Chicago, IL, USA). The survival rate of vitrified IVF embryos reached to as high as 97.6% (n = 42) 2 h post-warming, and hatching rate was 85.7% after 3 days culture in vitro. The data (Table 1) showed that there was no difference in Day 35 pregnancy rate among vitrified-double, vitrified-single, and fresh ET control groups. However, on Day 75 post-ET, there was a significantly higher fetal loss found in the vitrified-double transfer group (41.1%) compared to those of vitrified-single transfers (16.6%) and fresh-single group (11.9%) (P < 0.05). The pregnancy rate on Day 75 of 51.4% achieved with vitrified-single transfers was comparable to the 43.3% achieved with the fresh-single control transfers but was significantly higher than the 31.1% of the vitrified-double transfer group. This study demonstrated that double embryo transfers can aggravate high fetal loss and/or abortion when sexed IVF embryos are transferred, and ET with 1 sexed/vitrified embryo per recipient is sufficient to establish satisfactory pregnancy, comparable to that achieved with fresh embryos.

Table 1.Pregnancy and fetal loss of sexed/vitrified bovine IVF embryos following single and double transfers

Supported by USDA/CSREES-SBIR: 2006-03069 Phase II to F. Du.