Polymorphonuclear leukocyte migration through human amnion membrane

A new in vitro model has been developed for studying migration of human polymorphonuclear leukocytes (PMN) through living native cellular and matrix barriers. Human amnion membrane consists of a single layer of epithelium bound to a continuous basement membrane interfacing an avascular collagenous stroma. Living amnion was placed in plastic chambers with separate compartments on each side of the membrane. PMN were introduced on the epithelial side of the amnion, and a Millipore filter (Millipore Corp., Bedford, Mass.) was placed against the stromal side. In response to N-formylmethionyl-leucyl- phenylanlanine (FMLP) chemoattractant, PMN penetrated the full thickness of the amnion and were collected and counted on the filter. The rate of PMN traversal of the amnion was dependent on the concentration of FMLP (optimal at 10(-8)M) as well as the slope of the FMLP gradient across the amnion. The route of PMN migration was studied by transmission electron microscopy. PMN first attached to the epithelial surface, then infiltrated between intercellular junctions. PMN migrated around or through tight junction and hemidesmosome attachments. The PMN then penetrated the basement membrane and migrated through the dense collagenous stroma. The present amnion migration system has characteristics of the in vivo inflammatory state not described in any previous method for monitoring PMN migration in vitro. Prior methods have not used native epithelium, whole basement membrane, or collagenous stroma. PMN penetration of these barriers occurs in the normal inflammatory response and probably involves biochemical mechanisms not required for simple migration through the pores of an artificial filter. The amnion system can be useful for future biochemical and morphological studies of PMN penetration of these barriers and possible repair processes that may follow.     

Vergleichende Untersuchungen zum Kontaktverhalten verschiedener Arten der Gattung Tilapia (Cichlidae,Pisces) und ihrer Bastarde

• 1 Tilapia tholloni (Substratbrüter), T. nilotica (weiblicher Maulbrüter) und T. heudeloti macrocephala (männlicher Maulbrüter) wurden künstlich erbrütet und ihre angeborenen Kontaktreaktionen in standardisierten Attrappenversuchen untersucht.
• 2 Das Kontaktverhalten muß während einer kritischen Phase (bei T. nilotica unter den angegebenen Versuchsbedingungen bis ungefähr zum 21. Tag nach dem Ablaichen) aktiviert werden, wenn es längere Zeit andauern soll (= Reaktionsphase). Die Reaktionsphase kann bei T. nilotica mehrere Wochen dauern. In ihr nimmt die Reaktionsstärke (Anzahl und Dauer der Kontakte) zunächst rasch zu, erreicht ein Maximum und nimmt dann allmählich wieder ab. Anstieg, Maximum und Abnahme sind an bestimmte Entwicklungsabschnitte gebunden, weitgehend unabhängig davon, ob die Tiere zuvor schon Kontaktverhalten geäußert haben oder nicht. Die Kontaktreaktionen unterscheiden sich u. a. durch die Dauer der Kontakte: tholloni = 0,7 Sek., nilotica = 86,5 Sek., heudeloti m. = 1,5 Sek. je Tag und Tier (Maximalwerte bei bestimmten, für alle Arten gleichen Versuchsbedingungen).
• 3 Von der Aufzuchttemperatur hängt es ab, in welchem Entwicklungsabschnitt die Reaktionsphase liegt. Das Reaktionsmaximum junger T. nilotica lag bei 24° C am 9., bei 29° C am Tag nach der Eiablage.
• 4 Geblendete nilotica-Jungfische zeigten nur zu Beginn der Reaktionsphase schwaches Kontaktverhalten, normale Tiere äußerten gegenüber einer durchsichtigen Glasattrappe abgeschwächtes, nur kurze Zeit dauerndes Kontaktverhalten. Das Kontaktverhalten wird durch mechanische (Strömung) und optische Reize ausgelöter und gesteuert.
• 5 Junge Maulbrüter aus kleinen Eiern erreichten eine längere Kontaktdauer als solche aus größeren. Nach künstlicher Reduktion der Dottermenge um 10–20%) erhöhte sich die Kontaktdauer bei jungen nilotica um 23,1%, die Zahl der Kontakte nahm gleichzeitig um 7,6% ab. Die ♀♀ der substratbrütenden T. mariae legen große Eier, die Jungen zeigten gegenüber Attrappen intensive Kontaktreaktionen. Es wird die Frage diskutiert, ob das Erreichen einer bestimmten Eigröße eine Voraussetzung für das Entstehen des Kontaktverhaltens gewesen sein könnte.
• 6 Die Kurzkontakte junger T. tholloni werden aus ihrer Orientierungsreaktion abgeleitet und als Vorstufe des Kontaktverhaltens gedeutet.
• 7 7. Mehrere Tilapia-Arten wurden künstlich gekreuzt. Bei Verwendung von tholloni-Sperma wiesen die Bastarde eine erhöhte Sterblichkeit auf. Aus der Kreuzung T. tholloni ♀ ～ T. nilotica ♂ (Substratbrüter ～ weiblicher Maulbrüter) gingen nur ♀♀ hervor. Die Kreuzung T. heudeloti macrocephala ～ T. nilotica (männlicher ～ weiblicher Maulbrüter) erbrachte fertile F₁- und F₂-Generationen sowie alle vier möglichen Rückkreuzungen. Bei der Vererbung des Kontaktverhaltens (gemessen an der Dauer der Kontakte) scheinen relativ wenig Erbfaktoren mitzuwirken, T. heudeloti m. erwies sich gegenüber nilotica als praevalent. Ein Teil der Bastarde aus der Kreuzung T. tholloni ♀ ～ T. nilotica ♂ (F₁) (Substratbrüter ～ weiblicher Maulbrüter) lag auf der Merkmalsskala zwischen den Ausgangsarten, der Rest verteilte sich sowohl auf den Bereich von tholloni als auch auf den Bereich von nilotica.

     

Using functional groups to investigate community response to environmental changes: two grassland case studies

Functional groups (FG) are an useful generalization to investigate environmental change effects on biotic communities. Assigning species to FGs is a contextual task and carries an arbitrary element, regardless of whether the grouping is obtained a priori or by sophisticated numerical methods. Using two grassland community case studies, we show that even simple FG allocation based on growth form, architecture and longevity (plants and mosses), or foraging characteristics (above-ground invertebrates) can be useful to increase our understanding of community processes. For example, the sensitivity of organisms to climate change increases with trophic rank and is higher in disturbed than in undisturbed communities. Complexity of interaction webs (in terms of web connectance), however, is larger in undisturbed than in disturbed communities. A significant and important relationship is likely to exist between anthropogenic disturbances, community complexity and the ecosystem effects of climate change. Trophic interactions may be disrupted much easier by climate changes in disturbed than in undisturbed communities where complexity may be buffering these effects.     

Variations among cell lines in the synthesis of sphingolipids in de novo and recycling pathways

There are several pathways for the incorporation of sugars into glycosphingolipids (GSL). Sugars can be added to ceramide that contains sphinganine (dihydrosphingosine) synthesized de novo (pathway 1), to ceramide synthesized from sphingoid bases produced by hydrolysis of sphingolipids (pathway 2), and into GSL recycling from the endosomal pathway through the Golgi (pathway 3). We reported previously the surprising observation that SW13 cells, a human adrenal carcinoma cell line, synthesize most of their GSL in pathway 2. We now present data on the synthesis of GSL in four additional cell lines. Approximately 90% of sugar incorporation took place in pathway 2, and 10% or less in pathway 1, in human foreskin fibroblasts and NB41A3 neuroblastoma cells. In contrast, approximately 50-90% of sugar incorporation took place in pathway 1 in C2C12 myoblasts. The C2C12 cells divide more rapidly and synthesize 10-14 times as much GSL as the other three cell lines. In C6 glioma cells, approximately 30% of sugar incorporation occurred in pathway 1 and 60% in pathway 2. There was no relation between the utilization of pathways for GSL and sphingomyelin synthesis in foreskin fibroblasts and C2C12 cells. In both cells pathways 1 and 2 each accounted for 50% of incorporation of choline into sphingomyelin. In five of the six cell lines that we have studied, most GSL synthesis takes place in pathway 2. We suggest that when the need for synthesis is relatively low, as in slowly dividing cells, GSL are synthesized predominantly from sphingoid bases salvaged from the hydrolytic pathway. When cells are dividing more rapidly, the need for increased synthesis is met by upregulating the de novo pathway.      

Parallels,nonparallels, and plasticity in population differentiation of threespine stickleback within a lake

The frequent occurrence of parallel phenotypic divergence in similar habitats is often evoked when emphasizing the role of ecology in adaptive radiation and speciation. However, because phenotypic plasticity can contribute to the observed pattern of divergence, confirmation of divergence at loci underlying phenotypic traits is important for confirming adaptive divergence. In the present study, we examine parallel morphological, neutral, and potentially adaptive genetic divergence of threespine stickleback inhabiting different habitats within a lake. Three genetic clusters best explained the neutral genetic structure within the lake; however, morphological differences were only weakly connected to genetic clusters and there was considerable phenotypic variation within clusters. Among the factors that could contribute to the observed pattern of morphological and genetic divergence are phenotypic plasticity, selective mortality of hybrids, and habitat choice based on morphology. Several loci are identified as outliers indicating divergent selection between the morphs and some parallels in morphological and adaptive genetic divergence are found in stickleback spawning at two lava sites. However, neutral genetic structure indicates considerable genetic connectivity among the two lava sites, and the parallels in morphology may therefore represent selective distribution of phenotypes rather than parallel divergence. © 2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 98 , 803–813.     

Genetic structure of Anogeissus dhofarica (Combretaceae) populations endemic to the monsoonal fog oases of the southern Arabian Peninsula

Anogeissus dhofarica (Combretaceae) is an endemic tree of the monsoon affected coastal mountains of the southern Arabian Peninsula, being the character species of the Hybantho durae–Anogeissetum dhofaricae association, a drought deciduous, monsoon forest community found only in the Dhofar region of southern Oman and the eastern Al‐Mahra region of south‐east Yemen. Due to the steep precipitation gradient from the centre to the edges in this monsoon affected area, A. dhofarica is found in two different habitat types: in continuous woodland belts of the Hawf and Dhofar mountains, and in isolated, scattered woodland patches, as found especially in the Fartak Mts (south‐east Yemen). Fifteen populations (212 individuals) from across the whole distribution area of the species were analysed using amplified fragment length polymorphism fingerprinting to: (1) evaluate the consequences of population fragmentation on the genetic diversity harboured in isolated patches versus cohering stands of the species and (2) to reconstruct the phylogeographical pattern of A. dhofarica as a consequence of oscillations in the monsoon activity during the Pleistocene and Holocene. The analysis of among‐population genetic differentiation and within‐population genetic diversity in A. dhofarica populations resulted in a lack of genetic pauperization and genetic differentiation of populations of the distinctly isolated patches of the Fartak Mts compared to the more luxurious forests of the Hawf and Dhofar regions. This is considered to be due to the high buffer capacity against the loss of genetic diversity caused by the long‐lived life‐form of the species combined with the capability to propagate clonally and the relatively recent fragmentation of Anogeissus forests into the described patches rather than due to high values of gene flow among remnant populations caused by bee pollination and anemochorical and hydrochorical diaspore dispersal. The phylogeographical pattern of the species argues for a quite recent fragmentation of a once continuous forest belt of A. dhofarica that is rather connected with climate changes in the Holocene than triggered by aridity–humidity oscillations reported for the Pleistocene. © 2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 97 , 40–51.     

Xiphograptus and the evolution of virgella‐bearing graptoloids

Abstract: The virgellar spine is one of the most consistent features of the graptolite sicula. It is present in a large number of graptoloid groups, but evolved separately and independently in these as it is seen from the presence of the spine in either ventral (Axonophora) or dorsal (Phyllograptus, Xiphograptus) position. The evolution of the virgellar spine in the Pan‐Bireclinata in the Upper Dapingian to Lower Darriwilian time interval is known to follow four main steps, from a simple rutellum, through a lamelliform rutellum and a lanceolate virgella to the true virgellar spine. For the xiphograptids and in Phyllograptus, the origin and early development is less well documented, but appears to follow a similar path. However, the individual stages are condensed, and a true virgellar spine emerges already in the Floian time interval. A virgellar spine was found in Didymograptellus bifidus, necessitating a revision of the diagnosis of the genus Didymograptellus. A number of species of the virgellate genera Xiphograptus, Yutagraptus and Didymograptellus are described from isolated material for the first time. The species are useful for the biostratigraphic correlation of endemic mid‐continent North American faunas with the Pacific Type faunal realm. Xiphograptus artus sp. nov., Didymograptellus primus sp. nov. and Didymograptellus cowheadensis sp. nov. from the Cow Head Group of western Newfoundland are described as new.     

Characterization of novel microsatellite loci for red clover (Trifolium pratense L.) from enriched genomic libraries

Twenty‐seven polymorphic microsatellite markers were isolated from red clover (Trifolium pratense). Allelic variability and cross‐species amplification were assessed on 24 red clover and eight white clover (Trifolium repens) genotypes. The number of alleles detected in red clover ranged from two to 25. Observed and expected heterozygosities were high with average values of 0.71 and 0.88, respectively. Five of the 27 loci were also successfully amplified from white clover, where two to 13 alleles were detected. These highly polymorphic microsatellite loci provide powerful tools for population genetic studies as well as for marker‐assisted selection in this important forage legume species.     

Modeling brewers' yeast flocculation

Flocculation of yeast cells occurs during the fermentation of beer. Partway through the fermentation the cells become flocculent and start to form flocs. If the environmental conditions, such as medium composition and fluid velocities in the tank, are optimal, the flocs will grow in size large enough to settle. After settling of the main part of the yeast the green beer is left, containing only a small amount of yeast necessary for rest conversions during the next process step, the lagering. The physical process of flocculation is a dynamic equilibrium of floc formation and floc breakup resulting in a bimodal size distribution containing single cells and flocs. The floc size distribution and the single cell amount were measured under the different conditions that occur during full scale fermentation. Influences on flocculation such as floc strength, specific power input, and total number of yeast cells in suspension were studied. A flocculation model was developed, and the measured data used for validation. Yeast floc formation can be described with the collision theory assuming a constant collision efficiency. The breakup of flocs appears to occur mainly via two mechanisms, the splitting of flocs and the erosion of yeast cells from the floc surface. The splitting rate determines the average floc size and the erosion rate determines the number of single cells. Regarding the size of the flocs with respect to the scale of turbulence, only the viscous subrange needs to be considered. With the model, the floc size distribution and the number of single cells can be predicted at a certain point during the fermentation. For this, the bond strength between the cells, the fractal dimension of the yeast, the specific power input in the tank and the number of yeast cells that are in suspension in the tank have to be known. Copyright 1998 John Wiley & Sons, Inc.