Morphological and Seed Set characteristics of Centipedegrass accessions collected in China

Greater than two-fold differences were observed between the lowest and highest means for stolon number and length characteristics. A 60% difference in total length was observed between the longest stolon of accessions. There was a 42% difference between the accessions with the fastest and slowest growing stolons. Accession with the longest and shortest internode differed by 28%. Small differences were observed for leaf length and width. Seed set under selfing was less than 7% indicating high self-incompatibility. Open-pollinated seed set ranged from 37% to 87%. Coefficients of variation for the various characteristics measured indicate that the Chinese accessions should provide more variation for genetic improvement in this species. Data showed that measuring total stolon length at five weeks after transplanting was adequate for evaluating all of the stolon length and number characteristics in this study. The Chinese accession appear to be a valuable source of new germplasm for this species.     

Genetic analysis of the LEW.1AR1-iddm rat: an animal model for spontaneous diabetes mellitus

The LEW.1AR1-iddm/Ztm rat is a new animal model of type 1 diabetes mellitus, which shows an autosomal recessive mode of inheritance for the diabetes-inducing gene. The aim of this study was to define predisposing loci of the diabetic syndrome by linkage analysis using microsatellite markers. A backcross population of 218 rats (BN × LEW.1AR1-iddm) × LEW.1AR1-iddm was analyzed using 157 polymorphic microsatellite markers covering the entire genome. Three genomic regions showed a significant linkage to the diabetic syndrome. The first susceptibility locus on rat Chromosome (RNO) 1 (LOD score 4.13) mapped to the region 1q51–55, which codes for potential candidate genes like Ins1 and Nkx2-3. The second susceptibility locus was also localized on RNO1 in the centromeric region 1p11 (LOD score 2.7) encompassing the Sod2 gene. The third quantitative trait loci (LOD score 2.97) was located on RNO20 within the major histocompatibility complex region. Comparative mapping revealed that the homologous regions in the human genome contain the IDDM loci 1, 5, 8, and 17. The identification of diabetes susceptibility regions of the genetically uniform LEW.1AR1-iddm rat strain will pave the way toward a detailed characterization of the loci conferring diabetes development as well as their functional relevance for the pathogenesis of type 1 diabetes mellitus.     

Differential activation of CC chemokine receptors by AOP-RANTES

RANTES (regulated on activation normal T cell expressed) has been found at elevated levels in biological fluids from patients with a wide range of allergic and autoimmune diseases and is able to attract several subtypes of leukocytes including eosinophils and monocytes into inflamed tissue. Amino-terminal modifications of RANTES produce receptor antagonists which are candidates for blocking this cellular recruitment. Met-RANTES has been shown to modulate inflammation in vivo, while AOP-RANTES is a potent inhibitor of R5 human immunodeficiency virus type 1 (HIV-1) strains and has been shown to down-modulate CCR5 and prevent recycling of the receptor. We have studied the effect of AOP-RANTES in eosinophil activation and have found that it is able to efficiently elicit eosinophil effector functions through CCR3, as measured by the release of reactive oxygen species and calcium mobilization, whereas Met-RANTES is inactive in these assays. AOP-RANTES is found to inhibit CCR3-mediated HIV-1 infection with moderate potency, in contrast to its potent inhibition of CCR5-mediated HIV-1 infection. Furthermore, we have investigated the abilities of these modified proteins to down-modulate CCR1 and CCR3 from the surface of monocytes and eosinophils. We show here that AOP-RANTES is much less effective than RANTES in down-modulation of CCR1. Surprisingly, recycling of CCR1 was minimal after incubation with RANTES while there was complete recycling with AOP-RANTES. In the case of CCR3, no significant difference was found between RANTES and AOP-RANTES in down-modulation and recycling. It therefore appears that trafficking of RANTES receptors follows different patterns, which opens up potential new targets for therapeutic intervention.     

Characterization of synthetic C3a analog peptides on human eosinophils in comparison to the native complement component C3a

The C3a anaphylatoxin is a potent proinflammatory mediator derived from the complement system inducing biologic effects of human eosinophils like Ca2+ transients and the activation of the respiratory burst. These findings support an important role for C3a in diseases typically associated with a peripheral blood or tissue eosinophilia. Synthetic human C3a analogue peptides with variations at the C-terminal effector domain have been evaluated with respect to their binding affinity and signaling potency on human eosinophils. Flow cytometrical analysis and RT-PCR revealed that the C3a receptor is constitutively expressed on human eosinophils. Peptides bearing an N-terminal 9-fluorenylmethoxycarbonyl and the 6-aminohexanoyl motif were the most powerful peptides tested. Amino acid replacements in the conserved C-terminal pentapeptide decreased binding affinity and functional potency substantially. In addition, synthetic C3a analogue peptides induced C3aR internalization, led to transient changes of intracellular Ca2+ concentration, and did release reactive oxygen species in human eosinophils indicating the in vivo relevance of C3a-related sequences. The tripeptide LAR was found to be essential for C3a receptor binding on human eosinophils. Moreover, the putative binding motif of C3a anaphylatoxin is also crucial for the induction of biologic effects in the human system such as changes of intracellular Ca2+ concentration and the release of reactive oxygen species. This study demonstrates that the carboxyl terminus is important for the interaction with the C3aR and the biologic potency of C3a anaphylatoxin in the human system and plays a key role in the activation process of human eosinophils.     

Anomalous subdiffusion is a measure for cytoplasmic crowding in living cells

Macromolecular crowding dramatically affects cellular processes such as protein folding and assembly, regulation of metabolic pathways, and condensation of DNA. Despite increased attention, we still lack a definition for how crowded a heterogeneous environment is at the molecular scale and how this manifests in basic physical phenomena like diffusion. Here, we show by means of fluorescence correlation spectroscopy and computer simulations that crowding manifests itself through the emergence of anomalous subdiffusion of cytoplasmic macromolecules. In other words, the mean square displacement of a protein will grow less than linear in time and the degree of this anomality depends on the size and conformation of the traced particle and on the total protein concentration of the solution. We therefore propose that the anomality of the diffusion can be used as a quantifiable measure for the crowdedness of the cytoplasm at the molecular scale.     

Nestmate recognition in burying beetles: the "breeder's badge" as a cue used by females to distinguish their mates from male intruders

Burying beetles use small vertebrate carcasses as food for theirlarvae and defend these carcasses against intra- and interspecificcompetitors. Breeding associations on carcasses can consistof single females, heterosexual pairs, or various combinationsof males and females. When a heterosexual pair collaborate ina breeding attempt, they do not typically exhibit aggressivebehavior toward each other, but do attack newly arrived conspecificsthat attempt to usurp the carcass. We investigated the cuesinvolved in discrimination between breeding partners and intrudersby female burying beetles. We found that resident females toleratemales that have cared for a brood, as well as males that havenot cared for a brood but have been on a carcass for a day ortwo. Males that have had no prior contact with a carcass areattacked. Females appear to use a chemical cue, the "breeder'sbadge," an apolar substance on the male's cuticle that can beremoved by washing with pentane. This cue is reliably correlatedwith recent male experience with a carcass that is suitablefor reproduction. The breeder's badge develops as a result ofprolonged contact with such a carcass, and disappears on removalfrom the carcass; its presence does not require contact witha female or with larvae. Female recognition of their male partnersin burying beetles thus does not involve individual recognition,but rather recognition of reproductive condition.     

The Irving-Scholander legacy in polar physiology

The pioneer arctic and cold environment studies of Laurence Irving and Per Scholander, undertaken during the middle decades of the 20th century, have had a wide ranging and major influence on the direction and character of experimental research on polar species. Their investigations included comparative studies of metabolism, insulation, and acclimatization of mammals and birds in arctic Alaska and the tropics. Freezing, supercooling, and antifreeze research included fishes, insects, and plants. They examined the special problems of cooling in appendages of mammals and birds and the potential for acclimatization of these structures by repeated cold exposure. Studies of cold exposure in human populations considered possible adaptive reactions. Their research on diving mammals opened a vast new area for studies with attention to asphyxial tolerance and temperature regulation. A lesser-known accomplishment was an innovative approach to recovery of ancient atmospheric gases from polar ice fields, an approach that was stimulated by original investigations of frozen insect larvae and gas permeability through ice.     

Automated assignment of NOESY NMR spectra using a knowledge based method (KNOWNOE)

Automated assignment of NOESY spectra is a prerequisite for automated structure determination of biological macromolecules. With the program KNOWNOE we present a novel, knowledge based approach to this problem. KNOWNOE is devised to work directly with the experimental spectra without interference of an expert. Besides making use of routines already implemented in AUREMOL, it contains as a central part a knowledge driven Bayesian algorithm for solving ambiguities in the NOE assignments. These ambiguities mainly arise from chemical shift degeneration which allows multiple assignments of cross peaks. Using a set of 326 protein NMR structures, statistical tables in the form of atom-pairwise volume probability distributions (VPDs) were derived. VPDs for all assignment possibilities relevant to the assignments of interproton NOEs were calculated. With these data for a given cross peak with N possible assignments A _i(i = 1,...,N) the conditional probabilities P(A _i, a|V ₀) can be calculated that the assignment A _idetermines essentially all (a-times) of the cross peak volume V ₀. An assignment A _kwith a probability P(A _k, a|V ₀) higher than 0.8 is transiently considered as unambiguously assigned. With a list of unambiguously assigned peaks a set of structures is calculated. These structures are used as input for a next cycle of iteration where a distance threshold D _maxis dynamically reduced. The program KNOWNOE was tested on NOESY spectra of a medium size protein, the cold shock protein (TmCsp) from Thermotoga maritima. The results show that a high quality structure of this protein can be obtained by automated assignment of NOESY spectra which is at least as good as the structure obtained from manual data evaluation.