Post-dispersal seed removal in four Mediterranean oaks: species and microhabitat selection differ depending on large herbivore activity

Quercus forest regeneration is limited by different factors, post-dispersal predation being one of the most critical stages. We analysed seed removal of four coexisting Quercus species (Q. ilex, Q. suber, Q. faginea and Q. pyrenaica) in a Mediterranean forest located in Southern Spain. Marked and weighed acorns from each of the species were placed in experimental units with or without exclusion of large herbivores and in two microhabitat types (Q. ilex shade or open). Acorn removal was monitored for 120?days in order to test the effect of exclusion of large herbivore and microhabitat type on seed removal rates and species selection. Interestingly, the results of microhabitat and species selection differed depending on the presence of large herbivores. Removal was faster in sites without exclusion, where most seeds (??85%) disappeared during the first 9?days. In these sites, no differences in seed removal were found between different microhabitats, but seeds of two species, those with higher seed mass (Q. suber and Q. pyrenaica) were most preferred. However, under exclusion of large herbivores, seed removal was affected by the microhabitat, this being greater in Q. ilex shaded microhabitats, which showed a higher structural diversity. Also, species selection was completely different under exclusion of large herbivores, and seeds of Q. ilex and Q. faginea were removed faster. These results highlight the importance of large herbivore activity on seed removal and its effect on microhabitat and species selection. Therefore, specific selection by seed consumers may modify seedling recruitment and may have an important influence on the relative abundance of coexisting Quercus species.     

Hyaluronan receptors in the human ocular surface: a descriptive and comparative study of RHAMM and CD44 in tissues, cell lines and freshly collected samples

The purpose of this study was to demonstrate the presence of the receptor for hyaluronan-mediated motility (RHAMM) in human conjunctival epithelium and in two widely used cell lines from human corneal (HCE) and conjunctival (IOBA-NHC) epithelia. We compared the distribution of RHAMM proteins and mRNAs in human ocular surface tissues (corneal, limbal and conjunctival), HCE and IOBA-NHC cell lines, and corneal and conjunctival epithelia primary samples from healthy donors with the previously identified hyaluronan receptor CD44. We also aimed to determine if soluble CD44 (sCD44) was present in human tears, as it could have a role in the interaction of the tear fluid with hyaluronan. Protein expression was evaluated by Western blots and immunofluorescence microscopy. mRNA expression was evaluated by RT-PCR and Q-PCR. sCD44 was analyzed by ELISA in culture supernatants and in human tears. We describe the expression of RHAMM in human healthy conjunctiva and in HCE and IOBA-NHC cells at both protein and mRNA levels, and the presence of sCD44 in human tears. Furthermore, we detected CD44 and sCD44 expression variations in in vitro inflammatory conditions. This study also focused on the necessary caution with which the conclusions extracted from cell lines should be made, and in the great value of using primary samples as often as possible.     

Lipoteichoic acid is an important microbe-associated molecular pattern of Lactobacillus rhamnosus GG

Background

Receptors with a single transmembrane (TM) domain are essential for the signal transduction across the cell membrane. NMR spectroscopy is a powerful tool to study structure of the single TM domain. The expression and purification of a TM domain in Escherichia coli (E.coli) is challenging due to its small molecular weight. Although ketosteroid isomerase (KSI) is a commonly used affinity tag for expression and purification of short peptides, KSI tag needs to be removed with the toxic reagent cyanogen bromide (CNBr).

Result

The purification of the TM domain of p75 neurotrophin receptor using a KSI tag with the introduction of a thrombin cleavage site is described herein. The recombinant fusion protein was refolded into micelles and was cleaved with thrombin. Studies showed that purified protein could be used for structural study using NMR spectroscopy.

Conclusions

These results provide another strategy for obtaining a single TM domain for structural studies without using toxic chemical digestion or acid to remove the fusion tag. The purified TM domain of p75 neurotrophin receptor will be useful for structural studies.     

Regulation of 17β-hydroxysteroid dehydrogenases in cancer: regulating steroid receptor at pre-receptor stage

Increased intracelullar hormone concentration levels have been shown to be the cause of several endocrine-related cancers including breast, prostate, endometrial, ovarian, cervix, testicular, thyroid, and osteosarcoma. Deregulated expression of steroidogenic enzymes in these tumors seems to be the source of a positive balance in active steroids that bind to the corresponding nuclear receptor, thus ultimately stimulating cell proliferation. Among these enzymes, 17β-hydroxysteroid dehydrogenases catalyze the interconversion between 17-ketosteroids and 17-hydroxysteroids on the last steps of sex hormones biosynthesis and metabolism. To date, 14 isoforms have been identified in vertebrates although only 13 are present in humans. Development and clinical evaluation of specific inhibitors to block their activity is currently under progress especially against the best characterized members 1 to 5. Selectivity and potency of these drugs constitute the main challenge in this new approach to cancer and steroid-dependent diseases treatment at the "pre-receptor level". Here we review the current state of knowledge regarding the deregulation of the expression of some of these enzymes in endocrine-related tumors.     

Characterization of a Bacillus subtilis 64-kDa DNA polymerase X potentially involved in DNA repair

Bacillus subtilis gene yshC encodes a 64-kDa family X DNA polymerase (PolXBs), which contains all the critical residues involved in DNA and nucleotide binding as well as those responsible for catalysis of DNA polymerization, conserved in most family X members. Biochemical analyses of the purified enzyme indicate that PolXBs is a monomeric and strictly template-directed DNA polymerase, preferentially acting on DNA structures containing gaps from one to a few nucleotides and bearing a phosphate group at the 5' end of the downstream DNA. The fact that PolXBs is able to conduct filling of a single-nucleotide gap, allowing further sealing of the resulting nick by a DNA ligase, points to a putative role in base excision repair during the B. subtilis life cycle.     

Comparison of Methods for Estimating Bird Abundance and Trends From Historical Count Data

Abstract: The use of bird counts as indices has come under increasing scrutiny because assumptions concerning detection probabilities may not be met, but there also seems to be some resistance to use of model-based approaches to estimating abundance. We used data from the United States Forest Service, Southern Region bird monitoring program to compare several common approaches for estimating annual abundance or indices and population trends from point-count data. We compared indices of abundance estimated as annual means of counts and from a mixed-Poisson model to abundance estimates from a count-removal model with 3 time intervals and a distance model with 3 distance bands. We compared trend estimates calculated from an autoregressive, exponential model fit to annual abundance estimates from the above methods and also by estimating trend directly by treating year as a continuous covariate in the mixed-Poisson model. We produced estimates for 6 forest songbirds based on an average of 621 and 459 points in 2 physiographic areas from 1997 to 2004. There was strong evidence that detection probabilities varied among species and years. Nevertheless, there was good overall agreement across trend estimates from the 5 methods for 9 of 12 comparisons. In 3 of 12 comparisons, however, patterns in detection probabilities potentially confounded interpretation of uncorrected counts. Estimates of detection probabilities differed greatly between removal and distance models, likely because the methods estimated different components of detection probability and the data collection was not optimally designed for either method. Given that detection probabilities often vary among species, years, and observers investigators should address detection probability in their surveys, whether it be by estimation of probability of detection and abundance, estimation of effects of key covariates when modeling count as an index of abundance, or through design-based methods to standardize these effects.     

Localization of cholecystokinin-like immunoreactivity in the central nervous system of Triatoma infestans (Insecta: Heteroptera)

The distribution of cholecystokinin-like immunoreactivity was studied in the central nervous system of the heteropteran insect Triatoma infestans using high-sensitivity immunocytochemistry. In the protocerebrum, CCK-IR somata were observed in the anteromedial, anterolateral and posterior cell-body layers. The neuropils displayed different densities of immunoreactive neurites. Few immunoreactive somata were found in the optic lobe in both the medial and lateral soma rinds, as well as in the proximal optic lobe. Immunoreactive fibers were present in the medulla and lobula neuropils. The sensory deutocerebrum contained a higher number of immunopositive perikarya than the antennal mechanosensory and motor center. The antennal lobe glomeruli displayed a moderate density of immunoreactive fibers. With regard to the subesophageal ganglion, numerous CCK-IR somata were found close to the root of the mandibular nerve; others were present in the soma rind of the remaining neuromeres. CCK-IR perikarya were present in both thoracic ganglia, with the abdominal neuromeres containing the highest number of positive somata. The neuropils of both ganglia showed moderate densities of immunopositive processes. The distribution of CCK-LI in somata and neuropils of central nervous system of T. infestans is widespread suggesting that a CCK-like peptide may act mainly as a neuromodulator in the integration of information from distinct sensory receptors.     

Isolation and characterization of microsatellite markers for the ornamental discus fish Symphysodon discus and cross-species amplification in other Heroini cichlid species

The discus fishes (Symphysodon spp.) are economically important ornamental species. Thirteen microsatellite markers were developed from a CT(12) - and CA(12) -enriched whole genomic DNA library of Symphysodon discus. Allelic variability was tested on 44 individuals of two species (S. discus and S. aequifasciatus). Allelic richness ranged from two to 11 alleles per locus and observed heterozygosities from 0.083 to 0.998. All loci were at Hardy-Weinberg equilibrium, and no pair of loci showed linkage disequilibrium within a species. Cross-species amplification was also successfully performed in the Neotropical cichlids Uaru amphiacanthoides, Hoplarchus psittacus, Hypselecara coryphaenoides, Pterophyllum sp., Mesonauta sp. and Heros sp.     

拉萨河谷上段江孜沙棘Hippophea gyantsensis(Rousi) Lian种群的海拔梯度性变异

江孜沙棘[Hippophea gyantsensis(Rousi)Lian]是青藏高原特有的一种广生态幅的小乔木,在拉萨河谷地区的海拔3500~4200m范围内均有分布。前人工作多集中在江孜沙棘果实的开发利用方面,对其基础生态学研究较少。本研究旨在探讨江孜沙棘沿海拔梯度的群落组成和表型变异的规律。为此,在拉萨河谷上段沿海拔梯度由东向西设置了4个样带:3850m、3950m、4050m和4200m,每个样带设置2至3个10m×10m的样方进行研究。首先,详细记录了每个样方内林下维管植物的物种组成、样方内的沙棘盖度、海拔、样方与河岸的实际距离,并用DCA[detrended correspondence analysis(去势对应分析)]排序方法对群落及其组成物种进行排序分析。随机抽取了每个样方内的20个江孜沙棘植株个体,测定其胸径、基径、株高和叶片长度,用回归分析法分析这些变量和海拔之间的关系。研究结果表明,江孜沙棘在拉萨河谷内的主要生境分为4种类型,即:河边砾石滩地、河阶草滩、河边草甸和河边林缘,样方排序结果主要受海拔的影响;同时,江孜沙棘植株的基径、胸径和高度都随着海拔的升高而显著减小,而叶片长度与海拔之间无显著相关。本文研究结果表明,对江孜沙棘而言,海拔所代表的综合环境因子对其分布和表型有显著的影响,而局部光照可能也是影响其表型特征的重要生态因子。