Mediterranean views on epithelial polarity

Developmental and cell biologists viewed polarity through each other's eyes at the EMBO workshop on Epithelial Polarity in Development and Disease, March 27-31 2004, in Carry-le-Rouet, France, a small village west of Marseille on the rocky Mediterranean coast. The presentations highlighted our growing understanding, not only of the molecular mechanisms underlying polarity and the conservation of polarity complexes from worms to mammals, but also the diverse roles that epithelial polarity has during development.     

Differences between subcultures of the Mesorhizobium loti type strain from different culture collections

Because of differences in the reported 16S rRNA gene sequence of the Mesorhizobium loti type strain available from different culture collections, we collected different subcultures of this strain and compared them by 16S rDNA sequencing, SDS-PAGE of whole-cell protein extracts and RAPD-PCR. Our results indicate that the 16S rDNA sequence differences can be explained by the presence of two different organisms in one of the subcultures. In addition, even for subcultures of the type strain that had identical 16S rDNA sequences, small differences could be observed in the protein profiles and in the RAPD-PCR patterns. These latter observations indicate that maintenance procedures necessary for long-term preservation by freeze-drying can cause subcultures of the same original strain to undergo changes, effectively leading to different fingerprints even though 16S rDNA sequences remain identical.     

Exopolysaccharide (EPS) biosynthesis by Lactobacillus sakei 0-1: production kinetics, enzyme activities and EPS yields

AIMS: To determine optimal exopolysaccharide (EPS) production conditions of the mesophilic lactic acid bacterium strain Lactobacillus sakei 0-1 and to detect possible links between EPS yields and the activity of relevant enzymes. METHODS AND RESULTS: Fermentation experiments at different temperatures using either glucose or lactose were carried out. EPS production took place during the exponential growth phase. Low temperatures, applying glucose as carbohydrate source, resulted in the best bacterial growth, the highest amounts of EPS and the highest specific EPS production. Activities of 10 important enzymes involved in the EPS biosynthesis and the energy formation of Lact. sakei 0-1 were measured. The obtained results revealed that there is a clear link for some enzymes with EPS biosynthesis. It was also demonstrated clearly that the presence of rhamnose in the EPS building blocks is due to high activities of the enzymes involved in the rhamnose synthetic branch. CONCLUSION: EPS production in Lact. sakei 0-1 is growth-associated and displays primary metabolite kinetics. Glucose as carbohydrate source and low temperatures enhance the EPS production. The enzymes involved in the biosynthesis of the activated sugar nucleotides play a major role in determining the monomeric composition of the synthesized EPS. SIGNIFICANCE AND IMPACT OF THE STUDY: The proposed results contribute to a better understanding of the physiological factors influencing EPS production and the key enzymes involved in EPS biosynthesis by Lact. sakei.     

On local bifurcations in neural field models with transmission delays

Neural field models with transmission delays may be cast as abstract delay differential equations (DDE). The theory of dual semigroups (also called sun-star calculus) provides a natural framework for the analysis of a broad class of delay equations, among which DDE. In particular, it may be used advantageously for the investigation of stability and bifurcation of steady states. After introducing the neural field model in its basic functional analytic setting and discussing its spectral properties, we elaborate extensively an example and derive a characteristic equation. Under certain conditions the associated equilibrium may destabilise in a Hopf bifurcation. Furthermore, two Hopf curves may intersect in a double Hopf point in a two-dimensional parameter space. We provide general formulas for the corresponding critical normal form coefficients, evaluate these numerically and interpret the results.     

Integrated Community Profiling Indicates Long-Term Temporal Stability of the Predominant Faecal Microbiota in Captive Cheetahs

Understanding the symbiotic relationship between gut microbes and their animal host requires characterization of the core microbiota across populations and in time. Especially in captive populations of endangered wildlife species such as the cheetah (Acinonyx jubatus), this knowledge is a key element to enhance feeding strategies and reduce gastrointestinal disorders. In order to investigate the temporal stability of the intestinal microbiota in cheetahs under human care, we conducted a longitudinal study over a 3-year period with bimonthly faecal sampling of 5 cheetahs housed in two European zoos. For this purpose, an integrated 16S rRNA DGGE-clone library approach was used in combination with a series of real-time PCR assays. Our findings disclosed a stable faecal microbiota, beyond intestinal community variations that were detected between zoo sample sets or between animals. The core of this microbiota was dominated by members of Clostridium clusters I, XI and XIVa, with mean concentrations ranging from 7.5-9.2 log10 CFU/g faeces and with significant positive correlations between these clusters (P<0.05), and by Lactobacillaceae. Moving window analysis of DGGE profiles revealed 23.3-25.6% change between consecutive samples for four of the cheetahs. The fifth animal in the study suffered from intermediate episodes of vomiting and diarrhea during the monitoring period and exhibited remarkably more change (39.4%). This observation may reflect the temporary impact of perturbations such as the animal’s compromised health, antibiotic administration or a combination thereof, which temporarily altered the relative proportions of Clostridium clusters I and XIVa. In conclusion, this first long-term monitoring study of the faecal microbiota in feline strict carnivores not only reveals a remarkable compositional stability of this ecosystem, but also shows a qualitative and quantitative similarity in a defined set of faecal bacterial lineages across the five animals under study that may typify the core phylogenetic microbiome of cheetahs.     

Dietary Factors Associated with Faecal Consistency and Other Indicators of Gastrointestinal Health in the Captive Cheetah (Acinonyx jubatus)

Gastrointestinal diseases pose significant risks to captive cheetah survival and welfare. Multiple factors are thought to be associated with these diseases, but to date a comprehensive epidemiological survey of disease risk factors has not been conducted. A survey of diet and health parameters was completed for 184 captive cheetahs in 86 international facilities. Comparisons were made among dietary factors with respect to disease status and observed faecal consistency, incidence of vomiting and diarrhoea in the past 4 weeks. Extremely dry faeces were most common in cheetahs fed carcasses, but was still of low incidence (15%). Contrastingly, cheetahs fed commercially prepared diets had the highest prevalence of liquid faeces “always” or “often” (9%). Cheetahs fed raw meat diets had the highest prevalence of soft faeces with no shape (22%), as well as of firm and dry faeces (40%). No broad category of diet exerted any influence on the health parameters investigated. However, feeding of ribs at least once per week reduced the odds of diarrhoea (P = 0.020) and feeding of long bones (limbs) at least once per week was associated with a lower odds of vomiting (P = 0.008). Cheetahs fed muscle meat at least once per week had reduced odds of suffering from chronic gastritis (P = 0.005) or non-specific gastrointestinal disease (P < 0.001). The only factor identified as increasing the odds of chronic gastritis was feeding of horse “often” or “always” (P = 0.023). The findings of the current study build on existing empirical research to support a recommendation towards a greater inclusion of skeletal components. Current husbandry guidelines advocating the use of supplemented raw meat diets are likewise supported, but the use of horse meat, as well as commercially prepared diets for captive cheetahs, warrants caution until further research is conducted.     

Airways resistance and specific conductance for the diagnosis of obstructive airways diseases

Background

Airway resistance (R_AW) and specific airway conductance (sG_AW) are measures that reflect the patency of airways. Little is known of the variability of these measures between different lung diseases. This study investigated the contribution of R_AW and sG_AW to a diagnosis of obstructive airways disease and their role in differentiating asthma from COPD.

Methods

976 subjects admitted for the first time to a pulmonary practice in Belgium were included. Clinical diagnoses were based on complete pulmonary function tests and supported by investigations of physicians’ discretion. 651 subjects had a final diagnosis of obstructive diseases, 168 had another respiratory disease and 157 subjects had no respiratory disease (healthy controls).

Results

R_AW and sG_AW were significantly different (p < 0.0001) between obstructive and other groups. Abnormal R_AW and sG_AW were found in 39 % and 18 % of the population, respectively, in which 81 % and 90 % had diagnosed airway obstruction. Multiple regression revealed sG_AW to be a significant and independent predictor of an obstructive disorder. To differentiate asthma from COPD, R_AW was found to be more relevant and statistically significant. In asthma patients with normal FEV₁/FVC ratio, both R_AW and sG_AW were more specific than sensitive diagnostic tests in differentiating asthma from healthy subjects.

Conclusions

R_AW and sG_AW are significant factors that contribute to the diagnosis and differentiation of obstructive airways diseases.     

Vitamin D deficiency exacerbates COPD-like characteristics in the lungs of cigarette smoke-exposed mice

Background

Chronic obstructive pulmonary disease (COPD) is characterized by excessive inflammation and disturbed bacterial clearance in the airways. Although cigarette smoke (CS) exposure poses a major risk, vitamin D deficiency could potentially contribute to COPD progression. Many in vitro studies demonstrate important anti-inflammatory and antibacterial effects of vitamin D, but a direct contribution of vitamin D deficiency to COPD onset and disease progression has not been explored.

Methods

In the current study, we used a murine experimental model to investigate the combined effect of vitamin D deficiency and CS exposure on the development of COPD-like characteristics. Therefore, vitamin D deficient or control mice were exposed to CS or ambient air for a period of 6 (subacute) or 12 weeks (chronic). Besides lung function and structure measurements, we performed an in depth analysis of the size and composition of the cellular infiltrate in the airways and lung parenchyma and tested the ex vivo phagocytic and oxidative burst capacity of alveolar macrophages.

Results

Vitamin D deficient mice exhibited an accelerated lung function decline following CS exposure compared to control mice. Furthermore, early signs of emphysema were only observed in CS-exposed vitamin D deficient mice, which was accompanied by elevated levels of MMP-12 in the lung. Vitamin D deficient mice showed exacerbated infiltration of inflammatory cells in the airways and lung parenchyma after both subacute and chronic CS exposure compared to control mice. Furthermore, elevated levels of typical proinflammatory cytokines and chemokines could be detected in the bronchoalveolar lavage fluid (KC and TNF-α) and lung tissue (IP-10, MCP-1, IL-12) of CS-exposed vitamin D deficient mice compared to control mice. Finally, although CS greatly impaired the ex vivo phagocytic and oxidative burst function of alveolar macrophages, vitamin D deficient mice did not feature an additional defect.

Conclusions

Our data demonstrate that vitamin D deficiency both accelerates and aggravates the development of characteristic disease features of COPD. As vitamin D deficiency is highly prevalent, large randomized trials exploring effects of vitamin D supplementation on lung function decline and COPD onset are needed.

Electronic supplementary material

The online version of this article (doi:10.1186/s12931-015-0271-x) contains supplementary material, which is available to authorized users.