Eosinophil peroxidase produces hypobromous acid in the airways of stable asthmatics

Eosinophil peroxidase and myeloperoxidase use hydrogen peroxide to produce hypobromous acid and hypochlorous acid. These powerful oxidants may damage the lungs if they are produced as part of the inflammatory response in asthma. The aim of this study was to determine if peroxidases generate hypohalous acids in the airways of individuals with stable asthma, and if they affect lung function. Sputum was induced from patients with mild to moderate asthma and from healthy controls. Eosinophil peroxidase, myeloperoxidase, chlorinated and brominated tyrosyl residues, and protein carbonyls were measured in sputum supernatants. Eosinophil peroxidase protein was significantly elevated in asthmatic subjects whereas myeloperoxidase protein was not. There was significantly more 3-bromotyrosine (Br-Tyr) in proteins from the sputum of asthmatics compared to controls (0.79 vs. 0.23 mmol Br-Tyr/mol Tyr; medians p < .0001). Levels of 3-chlorotyrosine (0.23 vs. 0.14 mmol Cl-Tyr/mol Tyr; medians p = .11) and protein carbonyls (0.347 vs. 0.339 nmol/mg protein; medians p = .56) were not significantly increased in asthmatics. Levels of 3-bromotyrosine were strongly correlated with eosinophil peroxidase protein (r = 0.79, p < .0001). There were no significant correlations between the markers of oxidative stress and lung function. We conclude that eosinophil peroxidase produces substantial amounts of hypobromous acid in the airways of stable asthmatics. Although this highly reactive oxidant is a strong candidate for exacerbating inflammatory tissue damage in the lung, its role in asthma remains uncertain.     

The testicular sperm ducts and genital kidney of male Ambystoma maculatum (Amphibia,Urodela, Ambystomatidae)

The ducts associated with sperm transport from the testicular lobules to the Wolffian ducts in Ambystoma maculatum were examined with transmission electron microscopy. Based on the ultrastructure and historical precedence, new terminology for this network of ducts is proposed that better represents primary hypotheses of homology. Furthermore, the terminology proposed better characterizes the distinct regions of the sperm transport ducts in salamanders based on anatomy and should, therefore, lead to more accurate comparisons in the future. While developing the above ontology, we also tested the hypothesis that nephrons from the genital kidney are modified from those of the pelvic kidney due to the fact that the former nephrons function in sperm transport. Our ultrastructural analysis of the genital kidney supports this hypothesis, as the basal plasma membrane of distinct functional regions of the nephron (proximal convoluted tubule, distal convoluted tubule, and collecting tubule) appear less folded (indicating decreased surface area and reduced reabsorption efficiency) and the proximal convoluted tubule possesses ciliated epithelial cells along its entire length. Furthermore, visible luminal filtrate is absent from the nephrons of the genital kidney throughout their entire length. Thus, it appears that the nephrons of the genital kidney have reduced reabsorptive capacity and ciliated cells of the proximal convoluted tubule may increase the movement of immature sperm through the sperm transport ducts or aid in the mixing of seminal fluids within the ducts. © J. Morphol., 2013. © 2012 Wiley Periodicals, Inc.     

Exploitation of Mycobacterium tuberculosis Reporter Strains to Probe the Impact of Vaccination at Sites of Infection

Mycobacterium tuberculosis (Mtb) remains a major public health problem, with an effective vaccine continuing to prove elusive. Progress in vaccination strategies has been hampered by a lack of appreciation of the bacterium''s response to dynamic changes in the host immune environment. Here, we utilize reporter Mtb strains that respond to specific host immune stresses such as hypoxia and nitric oxide (hspX′::GFP), and phagosomal maturation (rv2390c′::GFP), to investigate vaccine-induced alterations in the environmental niche during experimental murine infections. While vaccination undoubtedly decreased bacterial burden, we found that it also appeared to accelerate Mtb''s adoption of a phenotype better equipped to survive in its host. We subsequently utilized a novel replication reporter strain of Mtb to demonstrate that, in addition to these alterations in host stress response, there is a decreased percentage of actively replicating Mtb in vaccinated hosts. This observation was supported by the differential sensitivity of recovered bacteria to the front-line drug isoniazid. Our study documents the natural history of the impact that vaccination has on Mtb''s physiology and replication and highlights the value of reporter Mtb strains for probing heterogeneous Mtb populations in the context of a complex, whole animal model.     

Histopathological and parasitological study of the gastrointestinal tract of dogs naturally infected with Leishmania infantum

Background

A nationwide survey on the microbial etiology of cases of subclinical mastitis in dairy cows was carried out on dairy farms in Sweden. The aim was to investigate the microbial panorama and the occurrence of antimicrobial resistance. Moreover, differences between newly infected cows and chronically infected cows were investigated.

Methods

In total, 583 quarter milk samples were collected from 583 dairy cows at 226 dairy farms from February 2008 to February 2009. The quarter milk samples were bacteriological investigated and scored using the California Mastitis Test. Staphylococci were tested for betalactamase production and presence of resistance was evaluated in all specific udder pathogens. Differences between newly infected cows and chronically infected cows were statistically investigated using logistic regression analysis.

Results

The most common isolates of 590 bacteriological diagnoses were Staphylococcus (S) aureus (19%) and coagulase-negative staphylococci (CNS; 16%) followed by Streptococcus (Str) dysgalactiae (9%), Str. uberis (8%), Escherichia (E.) coli (2.9%), and Streptococcus spp. (1.9%). Samples with no growth or contamination constituted 22% and 18% of the diagnoses, respectively. The distribution of the most commonly isolated bacteria considering only bacteriological positive samples were: S. aureus - 31%, CNS - 27%, Str. dysgalactiae - 15%, Str. uberis - 14%, E. coli - 4.8%, and Streptococcus spp. - 3.1%. There was an increased risk of finding S. aureus, Str. uberis or Str. dysgalactiae in milk samples from chronically infected cows compared to findings in milk samples from newly infected cows. Four percent of the S. aureus isolates and 35% of the CNS isolates were resistant to penicillin G. Overall, resistance to other antimicrobials than penicillin G was uncommon.

Conclusions

Staphylococcus aureus and CNS were the most frequently isolated pathogens and resistance to antimicrobials was rare.     

Development of indicators of ecosystem functioning in a temperate shelf sea: a combined fieldwork and modelling approach

A conceptual model of the main carbon and nitrogen flows through pelagic and benthic food webs was used to identify the key biogeochemical processes representing ecosystem functioning, and to select indicators of each of these processes. A combined fieldwork and modelling approach was used to provide the data required to evaluate the indicators in terms of their suitability for assessing and managing the impacts of climate change and demersal trawling. Four of our 16 proposed indicators (phytoplankton production and productivity, near-bed oxygen concentrations and oxygen penetration of the seabed) met the majority of criteria we used for evaluating indicators. Five indicators (depth of anoxic sediment, zoobenthos biomass, production, productivity and bioturbation potential) did not comply with sufficient criteria to be considered as good indicators. Six of our proposed indicators (zooplankton biomass, size structure, production and productivity; ecosystem productivity; ecosystem balance) could not be assessed for sensitivity and specificity using our models, and therefore need to be addressed in future work aimed at improving both the models and the fieldwork. Our results indicate that evaluation of indicators is difficult, because of the number and variety of human pressures which need to be considered in reality, and the interactions between these pressures and the ecosystem components which they affect. The challenge will be to establish if there are indeed any indicators which are able to meet the majority of criteria for good indicators in holistic ecosystem-based assessments.     

The phylogenetic distribution of the ampulla ureter and ampulla urogenital/uriniferous papilla in the Serpentes

The ampulla ureter and ampulla urogenital/uriniferous papilla represent differing morphologies of the caudal urogenital ducts in snakes. The ampulla ureter is an enlarged portion of the caudal extremity of the ureter that communicates the cranial regions of the ureter and the ductus deferens/Wolffian duct to the urodaeum. The ampulla urogenital/uriniferous papilla is an enlarged pouch, distinct from the ureter, which communicates the ureter and ductus deferens/Wolffian duct to the urodaeum. Although functional differences of these two structures are unknown, the ampulla urogenital/uriniferous papilla may have evolved for urine storage in males and females, and secondarily evolved a reproductive function in males. The most parsimonious optimization of the ampulla ureter and ampulla urogenital/uriniferous papilla indicates that the ampulla ureter is the ancestral state in snakes. Examining the presence or absence of the ampulla ureter and ampulla urogenital/uriniferous papilla in snakes on conflicting caenophidian phylogenies results in two hypotheses for the evolution of these variant morphologies: (1) The ampulla urogenital/uriniferous papilla evolved from the ampulla ureter independently in the Colubroidea and Elapoidea with subsequent losses of the ampulla urogenital/uriniferous papilla in the Elapoidea and (2) a single transition from the ampulla ureter to the ampulla urogenital/uriniferous papilla on the branch leading to the Colubroidea + Elapoidea with subsequent losses of the ampulla urogenital/uriniferous papilla in the Elapoidea and Colubroidea. The presence of the ampullae urogenital/uriniferous papilla in only the Colubroidea and Elapoidea highlights the affinity of these two taxonomic groups, a relationship that is strongly supported in published cladograms produced with molecular datasets.     

The reproductive biology of the depressed river mussel, Pseudanodonta complanata (Bivalvia: Unionidae), with implications for its conservation

Pseudanodonta complanata is listed as ‘Near Threatened’on the IUCN Red List (IUCN, 2006) and is a species of conservationpriority on the UK Biodiversity Action Plan. The UK is hostto some of the largest populations of this species, but littleis known about their reproductive biology. Two populations werestudied in the Great Ouse catchment and the Waveney and Yarecatchment, East Anglia, UK. Both populations are reproductivelyactive, producing viable glochidia. Small mussels (<30 mmlength) in both catchments indicate that recruitment is occurring.A short non-gravid period in May is followed by three monthsof glochidial formation in June, July and August; by Septembermussels contain mature glochidia ready for release the followingApril. Females brood between 5,000 and 50,000 glochidia, andthis scales with mussel length L (mm) as: n_glochL^2.1. The sexratio is skewed towards females (2.5 females:1 male), and malesare larger than females. No hermaphrodites were found duringthe histological examination of the gonads of 24 mussels. Thein vivo examination of demibranchs is shown to be an effectivenonsacrificial means of determining sex and gravidity. Conservationrecommendations include: minimizing management operations inriver stretches containing large populations; avoiding formsof management which preferentially remove large mussels (i.e.males and the most fecund females); performing management duringthe non-gravid period to avoid causing the premature releaseof glochidia; leaving sufficient time between management operationsfor populations to recover; and temporarily translocating musselsto refugia during management operations. (Received 9 February 2005; accepted 2 May 2007)     

Assessing lek attendance of male greater sage-grouse using fine-resolution GPS data: Implications for population monitoring of lek mating grouse

Counts of males displaying on breeding grounds are the primary management tool used to assess population trends in lekking grouse species. Despite the importance of male lek attendance (i.e., proportion of males on leks available for detection) influencing lek counts, patterns of within season and between season variability in attendance rates are not well understood. We used high-frequency global positioning system (GPS) telemetry data from male greater sage-grouse (Centrocercus urophasianus; n = 67) over five lekking seasons (2013–2017) at eight study sites in Nevada to estimate lek attendance rates. Specifically, we recorded daily locations of sage-grouse in relation to mapped lek boundaries and used generalized additive models to assess temporal variation in attendance rates by age class (subadult vs. adult). Average timing of peak attendance occurred on 16 April but varied from March 16, 2014 to April 21 , 2016. Overall, adult males attended leks at higher rates (0.683 at peak) and earlier in the season (19 March) than subadults (0.421 at peak on April 19). Peak attendance probability was positively related to cumulative winter precipitation. Daily probabilities of lek switching differed between adults (0.019 at peak on March 3) and subadults (0.046 at peak on March 22), and lek switching was negatively related to distance to nearest lek. Our results indicate variable patterns in lek attendance through time, and that lek switching may occur at higher rates than previously thought. We demonstrate the use of generalizable daily attendance curves to date-correct lek counts and derive estimates of male abundance, although such an approach will likely require the incorporation of information on age structure to produce robust results that are useful for population monitoring.