Expression and function of T cell homing molecules in Hodgkin’s lymphoma

Circulating T lymphocytes enter a tissue if they express appropriate chemokine receptors and adhesion molecules to engage ligands presented at this site. To aid rational development of T cell-based therapies for Hodgkin's lymphoma (HL), we have assessed the expression and function of homing receptors on tumour-infiltrating T cells in HL and compared them with T cells from unaffected lymph nodes and colorectal cancer tissue. Chemokine receptors CXCR3, CXCR4 and CCR7 were expressed on a large proportion of T cells within HL tissue and mediated chemotaxis to purified chemokine. The corresponding ligands (CXCL10, CXCL12, CCL21) were expressed on the malignant cells and/or vascular endothelium. Adhesion molecules including CD62L were widely expressed on HL-derived T cells and their corresponding ligands were detected on vessels within the tumour. This homing phenotype was distinct from T cells isolated from colorectal cancer, but matched closely the phenotype of T cells from unaffected lymph nodes. Thus, T cell recruitment to HL resembles entry of na?ve/central memory T cells into normal lymph nodes. This has important implications for current approaches to treat HL using T cells activated and expanded in vitro that lack CCR7 and CD62L expression.     

A Prospective Study of Transsulfuration Biomarkers in Autistic Disorders

The goal of this study was to evaluate transsulfuration metabolites in participants diagnosed with autism spectrum disorders (ASDs). Transsulfuration metabolites, including: plasma reduced glutathione (GSH), plasma oxidized glutathione (GSSG), plasma cysteine, plasma taurine, plasma sulfate, and plasma free sulfate among participants diagnosed with ASDs (n = 38) in comparison to age-matched neurotypical controls were prospectively evaluated. Testing was conducted using Vitamin Diagnostics, Inc. (CLIA-approved). Participants diagnosed with ASDs had significantly (P < 0.001) decreased plasma reduced GSH, plasma cysteine, plasma taurine, plasma sulfate, and plasma free sulfate relative to controls. By contrast, participants diagnosed with ASDs had significantly (P < 0.001) increased plasma GSSG relative to controls. The present observations are compatible with increased oxidative stress and a decreased detoxification capacity, particularly of mercury, in patients diagnosed with ASDs. Patients diagnosed with ASDs should be routinely tested to evaluate transsulfuration metabolites, and potential treatment protocols should be evaluated to potentially correct the transsulfuration abnormalities observed. An erratum to this article can be found at     

Comparative evaluation of Polymerase Chain Reaction-Restriction Enzyme Analysis (PRA) and sequencing of heat shock protein 65 (hsp65) gene for identification of aquatic mycobacteria

Traditional identification of mycobacteria based on cultural and biochemical tests can take several weeks and may fail to provide a precise identification. Polymerase Chain Reaction-Restriction Enzyme Analysis (PRA) of the gene encoding heat shock protein 65 kDa (hsp65) gene has been proposed as a rapid and inexpensive alternative approach. Despite being widely used for differentiation of mammalian mycobacteria, this method has only been applied in the identification of a small number of aquatic mycobacteria. The present study aimed to evaluate the potential use of PRA of hsp65 for the identification of aquatic mycobacteria compared with sequence analysis. Seventy one mycobacterial isolates including, 10 type/reference strains and the remainder field isolates, were subjected to PRA of a 441 bp fragment of this gene. For 68 representative isolates, sequence analysis was performed. All rapidly and slowly growing mycobacteria had best matches with 99.3% to 100% similarity with their corresponding species in the databanks. PRA proved to be a simple and rapid method for identifying aquatic mycobacteria. However, the incidence of similar or identical restriction patterns for some species of mycobacteria, and in particular, identification of new species of mycobacteria is a major problem using such a method. In contrast, the nucleic acid sequencing of the hsp65 gene yielded unambiguous results.     

Do tropical forest leaves suffer more insect herbivory? A comparison of tropical versus temperate herbivory,estimated from leaf litter

It is generally believed that tropical forests suffer more herbivory, as a proportion of leaf area, than do temperate forests. Reviews so far have compared studies performed by different authors using very different methodologies. Here we carried out studies on 125 samples at 86 localities in eastern North America and on 75 samples taken at five localities in Malaysia and Singapore, including both mature secondary and primary forest. Samples in North America were spread over 3 years. In tropical Asia, the samples were taken at four time slices at least 8 months apart, scattered over a 4-year period. Total herbivore damage during the lifetime of tree leaves was estimated from the percentage area damaged in recently fallen, undecayed leaves from the forest floor, using scanner-linked software. In terms of percentage damage per leaf, the results suggest that lowland tropical forest has significantly higher leaf herbivory (5.82%) than temperate forest (5.48%). This is in accord with the general expectation that aseasonal tropical forests should have more herbivory damage. However, when percentage damage ‘per unit time of growing season’ is calculated based on an estimate of leaf lifetime in the tropics, tropical lowland herbivory damage turns out to be a fraction (about one half) of that in the temperate zone. Thus, these results tend to put in question the widely held view that herbivore damage is markedly more intense in the tropics. Over total leaf lifetime, the intensity of damage in the tropical area is only slightly higher than temperate regions. In terms of intensity of herbivory on leaves per unit of time, the opposite seems to be the case. It is uncertain which index should be taken as more significant in interpreting the selection pressure for anti-herbivore defenses in the tropics.     

Solubilized phenyl-pyrazole ureas as potent, selective 5-HT2A inverse-agonists and their application as antiplatelet agents

Potent 5-HT_2A inverse-agonists containing phenyl-pyrazole ureas with an amino side chain were identified. Optimization of this series resulted in selective compounds that proved effective in modulating 5HT-induced amplification of ADP-stimulated human platelet aggregation.     

Banking North American buffalo semen

The purpose of this study was to develop a procedure to collect and preserve semen from wood bison (Bison bison athabascae) and plains bison (Bison bison bison). Semen samples from three wood and three plains bison bulls were collected by electroejaculation from June through October. In addition, sperm was collected from the cauda epididymis of seven plains bison. Semen was cryopreserved using two commercially available cryopreservation media, an egg yolk-based medium (Triladyl), and a medium free of products of animal origin (Andromed). Sperm morphology and motility were recorded on fresh and post-thawed semen samples. Total sperm motility was not different between plains and wood bison for the months of June (50%), July (69%) and October (54%). However, total sperm motility for wood bison was higher (P < 0.05) than plains bison for the months of August and September (August: 80% vs 55%; September: 73% vs 40%). Plains and wood bison did not differ in mean total and mean progressive motility (35 and 15%, respectively) of frozen-thawed sperm samples. The post-thaw motility of Triladyl-treated sperm was higher (P < 0.05) than Andromed-treated sperm (35% vs 13%, respectively). Interestingly, post-thawed epididymal spermatozoa had higher total motility (P < 0.05) than post-thawed electroejaculated sperm when cryopreserved with a medium free of products of animal origin (Andromed; 35% vs 9%, respectively). In conclusion, we used electroejaculation to collect high quality bison semen, and cryopreserved it for future needs.     

Efficient Degradation of Lignocellulosic Plant Biomass,without Pretreatment,by the Thermophilic Anaerobe “Anaerocellum thermophilum” DSM 6725

Very few cultivated microorganisms can degrade lignocellulosic biomass without chemical pretreatment. We show here that “Anaerocellum thermophilum” DSM 6725, an anaerobic bacterium that grows optimally at 75°C, efficiently utilizes various types of untreated plant biomass, as well as crystalline cellulose and xylan. These include hardwoods such as poplar, low-lignin grasses such as napier and Bermuda grasses, and high-lignin grasses such as switchgrass. The organism did not utilize only the soluble fraction of the untreated biomass, since insoluble plant biomass (as well as cellulose and xylan) obtained after washing at 75°C for 18 h also served as a growth substrate. The predominant end products from all growth substrates were hydrogen, acetate, and lactate. Glucose and cellobiose (on crystalline cellulose) and xylose and xylobiose (on xylan) also accumulated in the growth media during growth on the defined substrates but not during growth on the plant biomass. A. thermophilum DSM 6725 grew well on first- and second-spent biomass derived from poplar and switchgrass, where spent biomass is defined as the insoluble growth substrate recovered after the organism has reached late stationary phase. No evidence was found for the direct attachment of A. thermophilum DSM 6725 to the plant biomass. This organism differs from the closely related strain A. thermophilum Z-1320 in its ability to grow on xylose and pectin. Caldicellulosiruptor saccharolyticus DSM 8903 (optimum growth temperature, 70°C), a close relative of A. thermophilum DSM 6725, grew well on switchgrass but not on poplar, indicating a significant difference in the biomass-degrading abilities of these two otherwise very similar organisms.Utilization of lignocellulosic biomass derived from renewable plant material to produce ethanol and other fuels is viewed as a major alternative to petroleum-based energy sources (19). The efficient conversion of plant biomass to fermentable sugars remains a formidable challenge, however, due to the recalcitrance of the insoluble starting materials (13, 21, 36). Thermal and chemical pretreatments must be used to solubilize and release the sugars, but such processes are costly and not very efficient (17, 28). Most pretreatments utilize acids, alkali, or organic solvents (39). Moreover, the plant feedstocks vary considerably in their compositions. The main components of plant biomass and the sources of the fermentable sugars, cellulose and hemicellulose, are combined with lignin, which can occupy 20% (wt/wt) or more of the plant cell wall. The development of technologies to efficiently degrade plant biomass therefore faces considerable obstacles. The discovery or engineering of new microorganisms with the ability to convert the components of lignocellulosic biomass into sugars is therefore of high priority.Not many microorganisms are able to degrade pure crystalline cellulose, and the cellulose in plant biomass has a high order of crystallinity and is even less accessible to microbial or enzymatic attack (1, 12-14). Aerobic cellulolytic microorganisms usually secrete (hemi)cellulolytic enzymes containing carbohydrate-binding modules that serve to bind the catalytic domains to insoluble substrates. On the other hand, some anaerobic bacteria and fungi produce a large extracellular multienzyme complex called the cellulosome. This binds to and efficiently degrades cellulose and other polysaccharides, although it has a limited distribution in nature (3, 7). The rate at which microorganisms degrade cellulose increases dramatically with temperature (20), but the most thermophilic cellulosome-producing bacterium that has been characterized, Clostridium thermocellum, grows optimally near only 60°C (3, 9). A few anaerobic thermophiles are known that are able to grow on crystalline cellulose even though they lack cellulosomes, and in those cases the highest optimum growth temperature is 75°C (4, 32). Biomass conversion by thermophilic anaerobic microorganisms has many potential advantages over fermentation at lower temperatures. In particular, the organisms tend to have high rates of growth and metabolism, and the processes are less prone to contamination (30).The gram-positive bacterium “Anaerocellum thermophilum” strain Z-1320 is among the most thermophilic of the cellulolytic anaerobes (32). It grows optimally at 75°C at neutral pH and utilizes both simple and complex polysaccharides, although it does not grow on xylose or pectin (32). The end products of fermentation are lactate, ethanol, acetate, CO₂, and hydrogen. Although A. thermophilum Z-1320 grows very rapidly on crystalline cellulose (4), surprisingly, it has been studied very little since its discovery (32). We report here on the physiology of a very closely related strain, A. thermophilum DSM 6725, the genome of which was recently sequenced (16). The ability of A. thermophilum DSM 6725 to grow on different types of defined and complex substrates was investigated with a focus on switchgrass and poplar. These high-lignin plants have been selected as models for biomass-to-biofuel conversion by the BioEnergy Science Center (funded by the U.S. Department of Energy; http://bioenergycenter.org/). We show that A. thermophilum DSM 6725 is able to grow efficiently on both types of plant substrate without a chemical pretreatment step.     

Characterization of biocontrol traits in the entomopathogenic nematode Heterorhabditis georgiana (Kesha strain), and phylogenetic analysis of the nematode’s symbiotic bacteria

Our objective was to estimate the biocontrol potential of the recently discovered entomopathogenic nematode species Heterorhabditis georgiana (Kesha strain). Additionally, we conducted a phylogenetic characterization of the nematode’s symbiotic bacterium. In laboratory experiments, we compared H. georgiana to other entomopathogenic nematodes for virulence, environmental tolerance (to heat, desiccation, and cold), and host seeking ability. Virulence assays targeted Acheta domesticus, Agrotis ipsilon, Diaprepes abbreviatus, Musca domestica, Plodia interpunctella, Solenopsis invicta, and Tenebrio molitor. Each assay included H. georgiana and five or six of the following species: Heterorhabditis floridensis, Heterorhabditis indica, Heterorhabditis mexicana, Steinernema carpocapsae, Steinernema feltiae, Steinernema rarum, and Steinernema riobrave. Environmental tolerance assays included Heterorhabditis bacteriophora, H. georgiana, H. indica, S. carpocapsae, S. feltiae, and S. riobrave (except cold tolerance did not include S. carpocapsae or S. riobrave). Host seeking ability was assessed in H. bacteriophora, H. georgiana, S. carpocapsae, and Steinernema glaseri, all of which showed positive orientation to the host with S. glaseri having greater movement toward the host than S. carpocapsae (and the heterorhabditids being intermediate). Temperature range data (tested at 10, 13, 17, 25, 30 and 35 °C) indicated that H. georgiana can infect Galleria mellonella between 13 and 35 °C (with higher infection at 17–30 °C), and could reproduce between 17 and 30 °C (with higher nematode yields at 25 °C). Compared with other nematode species, H. georgiana expressed low or intermediate capabilities in all virulence and environmental tolerance assays indicating a relatively low biocontrol potential. Some novel observations resulted from comparisons among other species tested. In virulence assays, H. indica caused the highest mortality in P. interpunctella followed by S. riobrave; S. carpocapsae caused the highest mortality in A. domesticus followed by H. indica; and S. riobrave was the most virulent nematode to S. invicta. In cold tolerance, S. feltiae exhibited superior ability to cause mortality in G. mellonella (100%) at 10 °C, yet H. bacteriophora and H. georgiana exhibited the ability to produce attenuated infections at 10 °C, i.e., the infections resumed and produced mortality at 25 °C. In contrast, H. indica did not show an ability to cause attenuated infections. Based on the phylogenetic analysis, the bacterium associated with H. georgiana was identified as Photorhabdus luminescens akhurstii.