Assembly of binding loops on aromatic templates as VCAM‐1 mimetics

The design and synthesis of cyclic mimetics of VCAM‐1 protein that reproduce the integrin‐binding domain are presented. The unprotected peptide precursor 37 – 43 , Thr‐Gln‐Ile‐Asp‐Ser‐Pro‐Leu, was grafted onto functional templates of type naphthalene, biphenyl and benzyl through the chemoselective formation of C‐ and N‐terminal oximes resulting in a mixture of four isomeric forms due to syn–anti isomerism of the oxime bonds. Some isomers could be monitored by HPLC and identified by NMR. The molecule containing a naphthalene‐derived template was found to inhibit the VCAM‐1/VLA‐4 interaction more efficiently than previously reported for sulfur‐bridged cyclic peptides containing similar sequences. The finding confirms the importance of incorporating conformational constraints between the terminal ends of the peptide loop 37 – 43 in the design of synthetic inhibitors of the VCAM‐1/integrin interaction. Copyright © 1999 European Peptide Society and John Wiley & Sons, Ltd.     

Patch size and breeding status influence movement patterns in the threatened Malleefowl (Leipoa ocellata)

Information on the movement ecology of species can assist with identifying barriers to dispersal and appropriate management actions. We focus on the threatened Malleefowl (Leipoa ocellata) whose ability to move and disperse within fragmented landscapes is critical for their survival. We also investigate the possible effects of climate change on Malleefowl movement. We used solar-powered GPS telemetry to collect movement data and determine the influence of breeding status, remnant vegetation patches and environmental variables. Seven Malleefowl were tracked between 1 and 50 months, resulting in 20 932 fixes. While breeding, Malleefowl had significantly smaller home ranges (92 ± 43 ha breeding; 609 ± 708 ha non-breeding), moved shorter daily distances (1283 ± 605 breeding; 1567 ± 841 non-breeding) and stayed closer to the incubation mound (349 ± 324 m breeding; 3293 ± 2715 m non-breeding). Most Malleefowl effectively disassociated from the mound once breeding stopped, with two birds dispersing up to 10.2 km. Movement patterns were significantly correlated with the size of the remnant native vegetation patch, with smaller home ranges being utilized in small patches than in large patches. One male almost exclusively remained within a 107-ha patch for over 4 years, but a female crossed between closely spaced uncleared patches. Long-range movements of nearly 10 km daily displacement were recorded in large remnants almost exclusively when not breeding. Temperature and rain had a significant effect on movement: modelling suggests daily distances decline from 1.3 km at 25°C to 0.9 km at 45°C, with steeper declines over 30°C. The influence of patch size on movement patterns suggests that Malleefowl movement may be governed by the size of remnant patches and that habitat continuity may be important for facilitating recolonization after catastrophic events and maintaining genetic diversity. Climate change may reduce Malleefowl movement during hot, dry periods possibly affecting breeding success.     

Type IV pili are widespread among non-pathogenic Gram-positive gut bacteria with diverse carbohydrate utilization patterns

Type IV pili (T4P) are bacterial surface-exposed appendages that have been extensively studied in Gram-negative pathogenic bacteria. Despite recent sequencing efforts, little is known regarding these structures in non-pathogenic anaerobic Gram-positive species, particularly commensals of the mammalian gut. Early studies revealed that T4P in two ruminal Gram-positive species are associated with growth on cellulose, suggesting possible associations of T4P with substrate utilization patterns. In the present study, genome sequences of 118 taxonomically diverse, mainly Gram-positive, bacterial strains isolated from anaerobic (gastrointestinal) environments, have been analysed. The genes likely to be associated with T4P biogenesis were analysed and grouped according to T4P genetic organization. In parallel, consortia of Carbohydrate Active enZYmes (CAZymes) were also analysed and used to predict carbohydrate utilization abilities of selected strains. The predictive power of this approach was additionally confirmed by experimental assessment of substrate-related growth patterns of selected strains. Our analysis revealed that T4P systems with diverse genetic organization are widespread among Gram-positive anaerobic non-pathogenic bacteria isolated from different environments, belonging to two phylogenetically distantly related phyla: Firmicutes and Actinobacteria.     

A comparison of three methods for planning a census of Tawny Owl (Strix aluco) populations living at high territorial density

Censusing nocturnal species such as the Tawny Owl (Strix aluco) living in a dense population can lead to an overestimate if individual signatures are not available. A technique that separates the individualistic call of the Tawny Owl could be a useful tool for an exhaustive census. Six Tawny Owl males were followed for nine months; 654 vocalizations were analysed. Three methods – Spectrographic Cross Correlation (SPCC), Acoustic Space (AS) and Visual Spectrogram Comparison (VSC) – were tested to assess their ability to classify the typical male call. For SPCC, 10 randomly selected hoots for each male were compared: the distributions of correlation coefficients differed in only 26.7% of the cases when intra and inter individual variability are compared. For AS, all the spectrograms were measured through 13 parameters and intra and inter individual distances were compared: the interval containing 95.4% of intra-individual measures also contained 95.9% of inter-individual comparisons. Both SPCC and AS are considered not to be able to adequately separate subjects. For VSC, 31 randomly selected hoots were visually compared by 5 helpers; their classifications were compared pairwise and with the real situation; operators correctly identified a male in 70% of cases (mean = 70.4 SD = 5.4). If we integrate VSC with information coming from field notes, we have a more powerful tool than the mapping method. It is plausible that this technique can be useful for an exhaustive census of Tawny Owl populations living at high densities.     

Towards a physical description of habitat: quantifying environmental adversity (abiotic stress) in temperate forest and woodland ecosystems

1. Our aim was to develop a quantitative proxy for environmental adversity (abiotic stress) in temperate Eucalyptus and Nothofagus forest and woodland ecosystems.
2. Samples and measurements were collected at 42 sites across a rainfall gradient in southern Australia, an elevation gradient in south-eastern Australia, and a longitudinal transect (temperature gradient) in Patagonia, Argentina.
3. We compared the ability of (a) abiotic variables (14 soil and 21 climatic variables) and (b) the stable carbon isotope (δ¹³C) values of soil organic matter (SOM), to predict variation in leaf area index (LAI; a forest productivity variable).
4. The δ¹³C of SOM (soil aggregates) explained more variation (57%) in LAI than multivariate statistical models that integrated information on many abiotic variables. W* (a climatic water balance model) was also a powerful predictor variable, explaining 37% of the variability in LAI.
5 Synthesis . The stable carbon isotopic signature of soil aggregates is a powerful explanatory variable that may help us to quantify environmental adversity (abiotic stress) in temperate forest and woodland ecosystems.     

Correlation of C4ST-1 and ChGn-2 expression with chondroitin sulfate chain elongation in atherosclerosis

Subendothelial retention of lipoproteins by proteoglycans (PGs) is the initiating event in atherosclerosis. The elongation of chondroitin sulfate (CS) chains is associated with increased low-density lipoprotein (LDL) binding and progression of atherosclerosis. Recently, it has been shown that 2 Golgi enzymes, chondroitin 4-O-sulfotransferase-1 (C4ST-1) and chondroitin N-acetylgalactosaminyltransferase-2 (ChGn-2), play a critical role in CS chain elongation. However, the roles of C4ST-1 and ChGn-2 during the progression of atherosclerosis are not known. The aim of this study was to analyze the expression of C4ST-1 and ChGn-2 in atherosclerotic lesions in vivo and determine whether their expression correlated with CS chain elongation.Low-density lipoprotein receptor knockout (LDLr KO) mice were fed a western diet for 2, 4, and 8 weeks to stimulate development of atherosclerosis. The binding of LDL and CS PG in this mouse model was confirmed by chondroitinase ABC (ChABC) digestion and apolipoprotein B (apo B) staining. Gel filtration analysis revealed that the CS chains began to elongate as early as 2 weeks after beginning a western diet and continued as the atherosclerosis progressed. Furthermore, quantitative real-time polymerase chain reaction (qRT-PCR) showed that the mRNA levels of C4ST-1 and ChGn-2 increased after 8 weeks of this diet. In contrast, the mRNA levels of their homologs, C4ST-2 and ChGn-1, were unchanged. In addition, immunohistochemical analysis demonstrated that the expression of C4ST-1 and ChGn-2 appeared to have similar site-specific patterns and coincided with biglycan expression at the aortic root.Our results suggested that C4ST-1 and ChGn-2 may be involved in the elongation of CS chains in the arterial wall during the progression of atherosclerosis. Therefore, modulating their expression and activity might be a novel therapeutic strategy for atherosclerosis.     

Shunt in the diagnosis of initial lung lesion in smokers

Cigarette smoking is an important risk factor for all respiratory tract diseases. Unfortunately, the symptoms develop slowly, thus patients feel the consequences of the slowly developing inflammation too late. The inflammation first develops in the area of respiratory bronchioles. In this stage, the disease is asymptomatic. The study included a sample of 31 smokers, mean age 36.38 years, with normal spirometry indices, acid-base status and arterial blood gases. The mean smoking index was 11.28 smoking/years. All subjects were healthy, without any subjective health problems or disease indicators. The aim was to define dead lung area (V/Q) as an early indicator of changes in smokers. Study results demonstrated the mean shunt value in smokers of 8.25%, which showed positive correlation with smoking. The shunt size yielded negative correlation with the forced expiratory volume in one second and midexpiratory flow in smokers. In conclusion, determination of lung shunt is a simple method that is sensitive enough in the diagnosis of initial lung lesion due to cigarette smoking.     

Displacement of a mandibular incisor--an infrequent complication of mandibular fracture

A case of a mandibular fracture with an unusual complication is reported. A 13-year-old boy was admitted four years after conservative treatment of a symphyseal fracture. He complained of recurrent swelling. A radiographic evaluation showed a horizontally laid permanent mandibular left lateral incisor (PMLLI) that had probably slid into the fracture line and provoked repetitive infection episodes. After a surgery, a clinical and radiological analysis showed satisfactory healing.