THE REMOVAL OF BACTERIA FROM GLASS SURFACES WITH CALCIUM ALGINATE, GAUZE AND ABSORBENT COTTON WOOL SWABS

SUMMARY: Calcium alginate wool swabs were no more efficient or reliable than gauze or absorbent cotton wool swabs when tested on drinking glasses infected with known numbers of bacteria. The number of bacteria left on the glass after swabbing was determined by a modified roll tube method.     

Levels of IL-17F and IL-33 correlate with HLA-DR activation in clinical-grade human bone marrow–derived multipotent mesenchymal stromal cell expansion cultures

Background aims

Multipotent mesenchymal stromal cell (MSC)-based medicines are extensively investigated for use in regenerative medicine and immunotherapy applications. The International Society for Cell and Gene Therapy (ISCT) proposed a panel of cell surface molecules for MSC identification that includes human leukocyte antigen (HLA)-DR as a negative marker. However, its expression is largely unpredictable despite production under tightly controlled conditions and compliance with current Good Manufacturing Practices. Herein, we report the frequency of HLA-DR expression in 81 batches of clinical grade bone marrow (BM)-derived MSCs and investigated its impact on cell attributes and culture environment.

Mesenchymal stromal cell therapy: progress in manufacturing and assessments of potency

Mesenchymal stromal cell (MSC) therapies have been pursued for a broad spectrum of indications but mixed reports on clinical efficacy have given rise to some degree of skepticism regarding the effectiveness of this approach. However, recent reports of successful clinical outcomes and regulatory approvals for graft-versus-host disease, Crohn's disease and critical limb ischemia have prompted a shift in this perspective. With hundreds of clinical trials involving MSCs currently underway and an increasing demand for large-scale manufacturing protocols, there is a critical need to develop standards that can be applied to processing methods and to establish consensus assays for both MSC processing control and MSC product release. Reference materials and validated, uniformly applied tests for quality control of MSC products are needed. Here, we review recent developments in MSC manufacturing technologies, release testing and potency assays. We conclude that, although MSCs hold considerable promise clinically, economies of scale have yet to be achieved although numerous bioreactor technologies for scalable production of MSCs exist. Additionally, rigorous disease-specific product testing and comprehensive understanding of mechanisms of action, which are linked to relevant process and product release potency assays, will be required to ensure that these therapies continue to be successful.     

Impact of intentionally injected carbon dioxide hydrate on deep-sea benthic foraminiferal survival

Sequestration of carbon dioxide (CO₂) in the ocean is being considered as a feasible mechanism to mitigate the alarming rate in its atmospheric rise. Little is known, however, about how the resulting hypercapnia and ocean acidification may affect marine fauna. In an effort to understand better the protistan reaction to such an environmental perturbation, the survivorship of benthic foraminifera, which is a prevalent group of protists, was studied in response to deep-sea CO₂ release. The survival response of calcareous, agglutinated, and thecate foraminifera was determined in two experiments at ∼3.1 and 3.3 km water depth in Monterey Bay (California, USA). Approximately 5 weeks after initial seafloor CO₂ release, in situ incubations of the live–dead indicator CellTracker Green were executed within seafloor-emplaced pushcores. Experimental treatments included direct exposure to CO₂ hydrate, two levels of lesser exposure adjacent to CO₂ hydrate, and controls, which were far removed from the CO₂ hydrate release. Results indicate that survivorship rates of agglutinated and thecate foraminifera were not significantly impacted by direct exposure but the survivorship of calcareous foraminifera was significantly lower in direct exposure treatments compared with controls. Observations suggest that, if large scale CO₂ sequestration is enacted on the deep-sea floor, survival of two major groups of this prevalent protistan taxon will likely not be severely impacted, while calcareous foraminifera will face considerable challenges to maintain their benthic populations in areas directly exposed to CO₂ hydrate.     

Biological activity of aryl hydrocarbon receptor ligands in sediments from remote European lakes

1. Sediment extracts from 63 European remote lakes were tested for the presence of ligands of the aryl hydrocarbon receptor (AhR), using a yeast-based bioassay. The lakes belong to different European mountain regions in Central Norway, Scotland, Piedmont (Italy), Tyrol (Austria and Italy) and the Julian Alps (Slovenia).
2. AhR ligand activity varied about 40-fold among the examined lakes, with Scotland having the highest proportion of lakes with high AhR ligand activity, whereas samples with low AhR ligand activity predominate in the Julian Alps.
3. AhR ligand activity in sediment samples correlated with the concentration of several environmentally relevant compounds, including polycyclic aromatic hydrocarbons, hexachlorobenzene and several polychlorobiphenyl ether congeners. These correlations suggest an anthropogenic origin for the detected AhR ligands, rather than being generated from natural processes.
4. We conclude that pollutants deposited in remote lakes by atmospheric processes are active AhR ligands at the concentrations found in these lakes. Given the high toxicity associated with ectopic activation of AhR in different animal systems, this activity may represent a significant risk for the exposed fauna.     

Discovery of a large clonal patch of a social amoeba: implications for social evolution

Studies of genetic population structures of clonally reproducing macro-organisms have revealed large areas where only one clone is found. These areas, referred to as clonal patches, have not been shown to occur in free-living microbes until now. In free-living microbes, high genetic diversity at local scales is usually maintained by high rates of dispersal. We report, however, a highly dense, 12-m clonal patch of the social amoeba Dictyostelium discoideum in a cattle pasture located in a Texas Gulf Coast prairie. We confirm the presence of only one clone by the analysis of 65 samples and amplification of 10 polymorphic microsatellite loci. Samplings of additional cattle pastures nearby showed higher clonal diversity, but with a density of D. discoideum isolates lower than in the clonal patch. These findings show that high rates of microbial dispersal do not always produce genetic diversity at local scales, contrary to the findings of previous studies. The existence of clonal patches may be particularly important for microbial social evolution.     

The ovarian maturation cycle of the Norway lobster Nephrops norvegicus (Linnaeus, 1758) (Crustacea,Decapoda) from the western Mediterranean Sea

Summary

The Norway lobster Nephrops norvegicus lives in the continental margins of the western Mediterranean Sea at depths between 100 and 600 m. It constitutes an important fisheries resource and presents a seasonal reproductive pattern. Female Norway lobsters were obtained each month from a vessel fishing off Barcelona. One hundred females caught in June 2002 were kept in the laboratory. After spawning, ovarian samples were taken every 30 days with the objective of monitoring the first steps in ovarian maturation. The gonado-somatic index (GSI) remained low over the 6 months during which the females carried their eggs, plus for a further 2–3 months. However, this study suggests that ovarian maturation is a continuous process with two different phases, taking at least 6–8 months during which the female carries its eggs. There is an increase in oocyte numbers; the germinal zone produces oogonia; and the oocytes that develop migrate to the periphery, pushing the post-ovulatory follicles to the wall of the ovary and reinforcing it for subsequent spawning. Besides this increase in the number of oocytes, vitellogenesis begins 2–3 months after the eggs hatch. Oocytes then grow and the ovaries gain weight and change from a cream color to a blackish-green. When the GSI reached 10, spawning occurred and, from then on, the ovary is mainly composed of post-ovulatory follicles.     

METAMORPHOSIS OF THE LARVA OF THE POLYCHAETE ARENICOLA CRISTATA. MORPHOLOGY OF THE ‘HOUR-GLASS’ STAGE

The constricted ‘waist’ of the metamorphosing larva of the polychaete Arenicola cristata is described, using light and electron microscopy. The constriction is shown to be the consequence of the discharge and collapse of a post-trochal ring of epithelial cells which remain as functional components of the post-metamorphic juvenile. Morphological differentiation of neuro-effector and interneuronal contacts is initiated at this time. Muscular and neural changes are discussed in terms of their role in effecting metamorphosis.     

Conservation and versatility of a new set of primers for long‐PCR amplification of complete insect mitochondrial genomes based on Haematobia irritans mtDNA sequences

The amplification of complete mitochondrial genomes by long PCR (polymerase chain reaction) has been a major contribution to the large‐scale sequencing of arthropodan mitochondrial genomes. In this work, we designed six conserved long‐PCR primers to successfully recover the entire mitochondrial genome of the horn fly Haematobia irritans (Diptera: Muscidae) in two overlapping fragments. The conservation and versatility of these primers were tested for 17 other species from four major insect orders: Diptera (14), Coleoptera (1), Lepidoptera (1) and Hymenoptera (1). The amplification of complete mitochondrial genomes in orders other than Diptera suggested an even broader application of these primers, especially within the Hexapoda.     

New chromosome counts in the genus Cousinia and the related genus Schmalhausenia (Asteraceae, Cardueae)

Twenty chromosome counts are reported in the genus Cousinia and the monotypic genus Schmalhausenia , which are part of the Arctium group, from Armenia, Iran, Kazakhstan and Uzbekistan. Twelve are new and eight provide confirmation of scarce or disputable previous data. The correlation between karyological data, pollen type and molecular phylogeny is very close, and on this basis two main groups can be defined. One is the arctioid group, which comprises the genera Arctium and Schmalhausenia , and a small part of the genus Cousinia , with x  = 18. The other is the genus Cousinia s.s. , with a dysploid series ranging from x  = 13–11. Some considerations on the chromosomal evolution in the group are made.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Socety , 2003, 143 , 411–418.