Epratuzumab (humanised anti-CD22 antibody) in primary Sjögren's syndrome: an open-label phase I/II study

This open-label, phase I/II study investigated the safety and efficacy of epratuzumab, a humanised anti-CD22 monoclonal antibody, in the treatment of patients with active primary Sjögren's syndrome (pSS). Sixteen Caucasian patients (14 females/2 males, 33–72 years) were to receive 4 infusions of 360 mg/m² epratuzumab once every 2 weeks, with 6 months of follow-up. A composite endpoint involving the Schirmer-I test, unstimulated whole salivary flow, fatigue, erythrocyte sedimentation rate (ESR), and immunoglobulin G (IgG) was devised to provide a clinically meaningful assessment of response, defined as a ≥20% improvement in at least two of the aforementioned parameters, with ≥20% reduction in ESR and/or IgG considered as a single combined criterion. Fourteen patients received all infusions without significant reactions, 1 patient received 3, and another was discontinued due to a mild acute reaction after receiving a partial infusion. Three patients showed moderately elevated levels of Human anti-human (epratuzumab) antibody not associated with clinical manifestations. B-cell levels had mean reductions of 54% and 39% at 6 and 18 weeks, respectively, but T-cell levels, immunoglobulins, and routine safety laboratory tests did not change significantly. Fifty-three percent achieved a clinical response (at ≥20% improvement level) at 6 weeks, with 53%, 47%, and 67% responding at 10, 18, and 32 weeks, respectively. Approximately 40%–50% responded at the ≥30% level, while 10%–45% responded at the ≥50% level for 10–32 weeks. Additionally, statistically significant improvements were observed in fatigue, and patient and physician global assessments. Further, we determined that pSS patients have a CD22 over-expression in their peripheral B cells, which was downregulated by epratuzumab for at least 12 weeks after the therapy. Thus, epratuzumab appears to be a promising therapy in active pSS, suggesting that further studies be conducted.     

Seed addition experiments are more likely to increase recruitment in larger-seeded species

Colonisation theory predicts that large-seeded species are more likely to show increased seedling recruitment in response to seed addition than are small-seeded species. This is both because their seedlings tend to have better survivorship potential, and because their background density of germinating seedlings tends to be lower. We tested this hypothesis by combining data from a recent review of seed addition studies with seed mass data. Logistic regressions showed positive relationships between seed mass and propensity to increase seedling establishment in response to seed addition in experiments in which establishment success was assessed within 6 months or a year, but not in experiments in which establishment success was assessed more than a year after seed addition. When data for all time periods were combined, a generalised linear model including terms for seed mass, time and an interaction term showed a significant positive relationship between seed mass and species response to seed addition. Thus, knowing a species' seed mass significantly increased our ability to predict its response to seed supplementation.     

Identification of contractile vacuole proteins in Trypanosoma cruzi

Contractile vacuole complexes are critical components of cell volume regulation and have been shown to have other functional roles in several free-living protists. However, very little is known about the functions of the contractile vacuole complex of the parasite Trypanosoma cruzi, the etiologic agent of Chagas disease, other than a role in osmoregulation. Identification of the protein composition of these organelles is important for understanding their physiological roles. We applied a combined proteomic and bioinfomatic approach to identify proteins localized to the contractile vacuole. Proteomic analysis of a T. cruzi fraction enriched for contractile vacuoles and analyzed by one-dimensional gel electrophoresis and LC-MS/MS resulted in the addition of 109 newly detected proteins to the group of expressed proteins of epimastigotes. We also identified different peptides that map to at least 39 members of the dispersed gene family 1 (DGF-1) providing evidence that many members of this family are simultaneously expressed in epimastigotes. Of the proteins present in the fraction we selected several homologues with known localizations in contractile vacuoles of other organisms and others that we expected to be present in these vacuoles on the basis of their potential roles. We determined the localization of each by expression as GFP-fusion proteins or with specific antibodies. Six of these putative proteins (Rab11, Rab32, AP180, ATPase subunit B, VAMP1, and phosphate transporter) predominantly localized to the vacuole bladder. TcSNARE2.1, TcSNARE2.2, and calmodulin localized to the spongiome. Calmodulin was also cytosolic. Our results demonstrate the utility of combining subcellular fractionation, proteomic analysis, and bioinformatic approaches for localization of organellar proteins that are difficult to detect with whole cell methodologies. The CV localization of the proteins investigated revealed potential novel roles of these organelles in phosphate metabolism and provided information on the potential participation of adaptor protein complexes in their biogenesis.     

Asexual plants change just as often and just as fast as do sexual plants when introduced to a new range

Some of the world's most economically and environmentally damaging introduced species reproduce asexually. While sexually reproducing introduced herbs have proven capable of rapid evolution, no previous study has quantified morphological changes in multiple obligatory asexually reproducing introduced species, or asked whether their potential for change differs from that of sexual species. We measured herbarium specimens of eight asexual species introduced to Australia and/or New Zealand. We quantified changes in leaf area, leaf shape, leaf mass per area, plant height and internode length through time since the species’ introduction. Half of the asexual species demonstrated significant change in at least one trait. The observed changes were substantial – up to 561% per 100 years. The proportion of species showing at least one significant morphological change did not differ significantly between our asexual species, and data measured in the same manner for 34 sexually reproducing introduced species. There was also no significant difference in the rate of morphological change in sexual versus asexual species. That is, we found that sexual species have not changed more often, or more quickly than their asexual counterparts. Our results show that asexually reproducing species can undergo rapid morphological change at a rate matching that of sexually reproducing species when introduced to a new range. Our results also suggest that asexually reproducing plants may be able to respond to changing conditions better than was previously appreciated.     

Deep-sequencing of endothelial cells exposed to hypoxia reveals the complexity of known and novel microRNAs

In order to understand the role of microRNAs (miRNAs) in vascular physiopathology, we took advantage of deep-sequencing techniques to accurately and comprehensively profile the entire miRNA population expressed by endothelial cells exposed to hypoxia. SOLiD sequencing of small RNAs derived from human umbilical vein endothelial cells (HUVECs) exposed to 1% O₂ or normoxia for 24 h yielded more than 22 million reads per library. A customized bioinformatic pipeline identified more than 400 annotated microRNA/microRNA* species with a broad abundance range: miR-21 and miR-126 totaled almost 40% of all miRNAs. A complex repertoire of isomiRs was found, displaying also 5′ variations, potentially affecting target recognition. High-stringency bioinformatic analysis identified microRNA candidates, whose predicted pre-miRNAs folded into a stable hairpin. Validation of a subset by qPCR identified 18 high-confidence novel miRNAs as detectable in independent HUVEC cultures and associated to the RISC complex. The expression of two novel miRNAs was significantly down-modulated by hypoxia, while miR-210 was significantly induced. Gene ontology analysis of their predicted targets revealed a significant association to hypoxia-inducible factor signaling, cardiovascular diseases, and cancer. Overexpression of the novel miRNAs in hypoxic endothelial cells affected cell growth and confirmed the biological relevance of their down-modulation. In conclusion, deep-sequencing accurately profiled known, variant, and novel microRNAs expressed by endothelial cells in normoxia and hypoxia.     

Effects of age,replicative lifespan and growth rate of human nucleus pulposus cells on selecting age range for cell-based biological therapies for degenerative disc diseases

Autologous disc cell implantation, growth factors and gene therapy appear to be promising therapies for disc regeneration. Unfortunately, the replicative lifespan and growth kinetics of human nucleus pulposus (NP) cells related to host age are unclear. We investigated the potential relations among age, replicative lifespan and growth rate of NP cells, and determined the age range that is suitable for cell-based biological therapies for degenerative disc diseases. We used NP tissues classified by decade into five age groups: 30s, 40s, 50s, 60s and 70s. The mean cumulative population doubling level (PDL) and population doubling rate (PDR) of NP cells were assessed by decade. We also investigated correlations between cumulative PDL and age, and between PDR and age. The mean cumulative PDL and PDR decreased significantly in patients in their 60s. The mean cumulative PDL and PDR in the younger groups (30s, 40s and 50s) were significantly higher than those in the older groups (60s and 70s). There also were significant negative correlations between cumulative PDL and age, and between PDR and age. We found that the replicative lifespan and growth rate of human NP cells decreased with age. The replicative potential of NP cells decreased significantly in patients 60 years old and older. Young individuals less than 60 years old may be suitable candidates for NP cell-based biological therapies for treating degenerative disc diseases.     

Latitude, seed predation and seed mass

Aim We set out to test the hypothesis that rates of pre‐ and post‐dispersal seed predation would be higher towards the tropics, across a broad range of species from around the world. We also aimed to quantify the slope and predictive power of the relationship between seed mass and latitude both within and across species. Methods Seed mass, pre‐dispersal seed predation and post‐dispersal seed removal data were compiled from the literature. Wherever possible, these data were combined with information regarding the latitude at which the data were collected. Analyses were performed using both cross‐species and phylogenetic regressions. Results Contrary to expectations, we found no significant relationship between seed predation and latitude (log₁₀ proportion of seeds surviving predispersal seed predation vs. latitude, P = 0.63; R² = 0.02; n = 122 species: log₁₀ proportion of seeds remaining after postdispersal seed removal vs. latitude, P = 0.54; R² = 0.02; n = 205 species). These relationships remained non‐significant after variation because of seed mass was accounted for. We also found a very substantial (R² = 0.21) relationship between seed mass and latitude across 2706 species, with seed mass being significantly higher towards the tropics. Within‐species seed mass decline with latitude was significant, but only about two‐sevenths, as rapid as the cross‐species decline with latitude. Results of phylogenetic analyses were very similar to cross‐species analyses. We also demonstrated a positive relationship between seed mass and development time across ten species from dry sclerophyll woodland in Sydney (P < 0.001; R² = 0.77; Standardized Major Axis slope = 0.14). These data lend support to the hypothesis that growing period might affect the maximum attainable seed mass in a given environment. Main conclusions There was no evidence that seed predation is higher towards the tropics. The strong relationship between seed mass and latitude shown here had been observed in previous studies, but had not previously been quantified at a global scale. There was a tenfold reduction in mean seed mass for every c. 23° moved towards the poles, despite a wide range of seed mass within each latitude.     

Does a latitudinal gradient in seedling survival favour larger seeds in the tropics?

The mean size of seeds produced by plants at the equator is two to three orders of magnitude higher than the mean size of seeds produced by plants at 60°. We compiled data from the literature to assess the possibility that this latitudinal gradient in seed size allows species to cope with more difficult seedling establishment conditions in tropical environments. We found no relationship between latitude and seedling survival through 1 week (P = 0.27, n = 112 species). There was also no evidence that a larger seed mass is required to gain a given level of seedling survival in tropical environments (P = 0.37, n = 112 species), and no relationship between latitude and the duration of the juvenile period (P = 0.57, n = 132 species). Thus, our results are not compatible with the idea that seedling establishment is more difficult in the tropics.