Single-cell mass cytometry of differential immune and drug responses across a human hematopoietic continuum

Flow cytometry is an essential tool for dissecting the functional complexity of hematopoiesis. We used single-cell "mass cytometry" to examine healthy human bone marrow, measuring 34 parameters simultaneously in single cells (binding of 31 antibodies, viability, DNA content, and relative cell size). The signaling behavior of cell subsets spanning a defined hematopoietic hierarchy was monitored with 18 simultaneous markers of functional signaling states perturbed by a set of ex vivo stimuli and inhibitors. The data set allowed for an algorithmically driven assembly of related cell types defined by surface antigen expression, providing a superimposable map of cell signaling responses in combination with drug inhibition. Visualized in this manner, the analysis revealed previously unappreciated instances of both precise signaling responses that were bounded within conventionally defined cell subsets and more continuous phosphorylation responses that crossed cell population boundaries in unexpected manners yet tracked closely with cellular phenotype. Collectively, such single-cell analyses provide system-wide views of immune signaling in healthy human hematopoiesis, against which drug action and disease can be compared for mechanistic studies and pharmacologic intervention.     

A high-resolution paleoclimate record spanning the past 25,000 years in southern East Africa

High-resolution profiles of the mass accumulation rate of biogenic silica and other geochemical proxies in two piston cores from northern Lake Malawi provide a climate signal for this part of tropical Africa spanning the past 25,000 years. The biogenic silica mass accumulation rate was low during the relatively dry late Pleistocene, when the river flux of silica to the lake was suppressed. Millennial-scale fluctuations, due to upwelling intensity, in the late Pleistocene climate of the Lake Malawi basin appear to have been closely linked to the Northern Hemisphere climate until 11 thousand years ago. Relatively cold conditions in the Northern Hemisphere coincided with more frequent north winds over the Malawi basin, perhaps resulting from a more southward migration of the Intertropical Convergence Zone.     

Associations between antimicrobial use and the prevalence of antimicrobial resistance in fecal Escherichia coli from feedlot cattle in western Canada

A randomized, controlled, blinded clinical trial was performed at a research feedlot in western Canada. Auction-market-derived steers (n = 288) were randomly assigned to 1 of 3 treatments: 1) no antimicrobials on arrival; 2) oxytetracycline in the starter ration for 14 d; and 3) long-acting oxytetracycline subcutaneously on day 0. Minimal inhibitory concentrations of 7 antimicrobials were determined for 3 generic fecal E. coli isolates per animal on arrival and throughout the feeding period. There was a low prevalence of antimicrobial resistance in generic E. coli isolates from calves on arrival. There were increased proportions of cattle with resistant E. coli isolates early in the feeding period among calves in groups 2 and 3. Individual animal treatments were not associated with increased proportions of cattle with resistant E. coli isolates preslaughter. There was no difference in the proportion of animals with E. coli isolates resistant to tetracycline between the treatment groups preslaughter. However, there were significantly more animals with tetracycline resistant isolates of E. coli preslaughter than at arrival.     

Repellent effects of five monoterpenoid odours against Tribolium castaneum (Herbst) and Rhyzopertha dominica (F.) in Calabar, Nigeria

Secondary plant compounds are recognised as important components of plant defence system against herbivores and pathogens. Five monoterpenoids, (R)-linalool, 1,8-cineole, (S)-2-heptyl acetate, (S)-2-heptanol and citral, which are natural components of the essential oils of Aframomum melegueta (K. Schum) and Zingiber officinale (Roscoe), were tested at the ratios in which they occur naturally for repellent activity against Tribolium castaneum (Herbst.) and Rhyzopertha dominica (F.) in a 4-way olfactometer. The results showed the repellent properties of the compounds as both beetles spent less time in the olfactometer arm containing the test stimuli. (R)-linalool and (S)-2-heptanol were stronger repellent compounds than the others. Linalool showed good repellent activity against T. castaneum (P = 0.001) as the insect spent 1.22 min in the test arm compared to the control arms (2.78 min), and R. dominica (P = 0.001) with 0.89 min in the test arm compared to 2.87 min in the control arms. With (S)-2-heptanol, T. castaneum spent 1.23 min in the test arm compared to 2.83 min in the control arms. R. dominica spent 1.61 min in the test arm and 2.69 min in the control arms. For the number of entries or visits made, while both insects were significantly repelled (P < 0.05) by the linalool-treated arm than the control, only R. dominica was repelled by the (S)-2-heptanol-treated arm (P = 0.038) compared to the control arms. The results indicate that A. melegueta and Z. officinale essential oils and their components could be suitable as safer repellents or fumigants against T. castaneum and R. dominica.     

Breed-associated phenotypes in canine atopic dermatitis

Canine atopic dermatitis is a multifaceted disease, whose clinical presentation may be affected by numerous factors, including the genetic background of the animal, the environment, the offending allergens and flare factors. In particular, breed-associated differences have often been mentioned but never defined precisely. Using a large data set of atopic dogs, we document in this study the clinical presentation of nine often-affected breeds and demonstrate the existence of substantial differences between the clinical phenotype of each breed and the whole population. Some of the differences may be due to genetic differences while others are most likely to be associated with variations in environmental factors.     

Low-Molecular-Weight Fucoidan Induces Endothelial Cell Migration via the PI3K/AKT Pathway and Modulates the Transcription of Genes Involved in Angiogenesis

Low-molecular-weight fucoidan (LMWF) is a sulfated polysaccharide extracted from brown seaweed that presents antithrombotic and pro-angiogenic properties. However, its mechanism of action is not well-characterized. Here, we studied the effects of LMWF on cell signaling and whole genome expression in human umbilical vein endothelial cells and endothelial colony forming cells. We observed that LMWF and vascular endothelial growth factor had synergistic effects on cell signaling, and more interestingly that LMWF by itself, in the absence of other growth factors, was able to trigger the activation of the PI3K/AKT pathway, which plays a crucial role in angiogenesis and vasculogenesis. We also observed that the effects of LMWF on cell migration were PI3K/AKT-dependent and that LMWF modulated the expression of genes involved at different levels of the neovessel formation process, such as cell migration and cytoskeleton organization, cell mobilization and homing. This provides a better understanding of LMWF’s mechanism of action and confirms that it could be an interesting therapeutic approach for vascular repair.     

Genetic Aspects of Reproduction in Sheep

Maintenance of high levels of realized fertility (defined as the percentage of ewes that lamb) and appropriate levels of fecundity are critical for efficient sheep production. The optimal level of fecundity in most situations is well below the maximum attainable level and can be targeted by combining selection among and within breeds with use of an expanding array of single-gene mutations affecting ovulation rate and litter size. The heritability of litter size is approximately 0.10, allowing changes of up to 2%/year from simple mass selection. Mutations in several genes associated with the transforming growth factor β superfamily ( BMPRIB , GDF9 and sex-linked BMP15 ) can increase ovulation rates by 0.7–1.5 ova in heterozygous ewes. However, ewes that are homozygous for BMP15 or GDF9 mutations are sterile, so use of these mutations requires carefully structured breeding programmes. Improvements in fertility may be critical for autumn lambing or programmes that aspire to lamb throughout the year. Selection to improve fertility in spring matings has been successful; selected adult ewes have lambing rates of 80–85% in October and early November. The selected ewes have a dramatically reduced seasonal anestrus, and many ewes continue to cycle during spring and summer. Major genes affecting seasonal breeding have not been identified in sheep. Polymorphisms in the melatonin receptor 1a gene appear to be associated with seasonal breeding in some, but not all breeds. However, functional genomic studies of genes associated with circadian and circannual rhythms have potential to reveal additional candidate genes involved in seasonal breeding.     

Establishment of diagnostic criteria for feline nonflea-induced hypersensitivity dermatitis

Hypersensitivity dermatitides (HD) are commonly seen in cats, and they are usually caused by environmental, food and/or flea allergens. Affected cats normally present with one of the following clinical reaction patterns: head and neck excoriations, usually symmetrical self-induced alopecia, eosinophilic skin lesions or miliary dermatitis. Importantly, none of these clinical presentations is considered to be pathognomonic for HD skin diseases, and the diagnosis of HD is usually based on the exclusion of other pruritic diseases and on a positive response to therapy. The objectives of this study were to propose sets of criteria for the diagnosis of nonflea-induced HD (NFHD). We recruited 501 cats with pruritus and skin lesions and compared clinical parameters between cats with NFHD (encompassing those with nonflea, nonfood HD and those with food HD), flea HD and other pruritic conditions. Using simulated annealing techniques, we established two sets of proposed criteria for the following two different clinical situations: (i) the diagnosis of NFHD in a population of pruritic cats; and (ii) the diagnosis of NFHD after exclusion of cats with flea HD. These criteria sets were associated with good sensitivity and specificity and may be useful for homogeneity of enrolment in clinical trials and to evaluate the probability of diagnosis of NFHD in clinical practice. Finally, these criteria were not useful to differentiate cats with NFHD from those with food HD.     

Dereplication and Chemotaxonomical Studies of Marine Algae of the Ochrophyta and Rhodophyta Phyla

Dereplication and chemotaxonomic studies of six marine algae of the Ochrophyta and one of the Rhodophyta phyla resulted in the detection of 22 separate compounds. All 16 secondary metabolites, including four new compounds (16–19), could be rapidly dereplicated using HPLC-NMR and HPLC-MS methodologies in conjunction with the MarinLit database. This study highlights the advantages of using NMR data (acquired via HPLC-NMR) for database searching and for the overall dereplication of natural products.