Pathological changes in the nephrocytes of the shore crab, Carcinus maenas, following injection of bacteria

The gills of Carcinus maenas were examined by light and electron microscopy following injection of either sterile saline or the bacteria Bacillus cereus and Moraxella sp., to determine any role(s) for the nephrocytes in the host defense reactions. The results showed that although intact bacteria were not sequestered to the nephrocytes, these cells were active in the removal of large quantities of cell debris from the hemolymph. Much of this material was derived from the breakdown of the hemocytes in response to the presence of bacteria and it's accumulation in the central vacuoles of the nephrocytes resulted in the degradation of these cells. It is proposed that while nephrocytes do not phagocytose intact bacteria, they augment the host defenses by clearing much of the hemocyte and associated bacterial debris from the gills, thus preventing blockage of the lamellar sinuses and subsequent impairment of respiration.     

Cellular defense reactions of the shore crab, Carcinus maenas: in vivo hemocytic and histopathological responses to injected bacteria

The cellular defense reactions of the shore crab, Carcinus maenas, were studied, following injections of the bacteria Bacillus cereus and Moraxella sp., by histological and ultrastructural examination of the gills, heart, and hepatopancreas. The majority of the bacteria were sequestered to the gills, but some were also later evident in the heart and hepatopancreas. The presence of the bacteria in the gills initiated the formation of numerous small cell clumps, composed of both refractile and phagocytic cells, which entrapped many microorganisms. The clumps reached a maximum size 6 hr after inoculation and although some were cleared from the gills others persisted for 7 days, becoming more compact and necrotic during this period. Clump formation appears to occur following recognition of the bacteria as foreign and results in the hemocytes becoming sticky and adherent. The response is very effective in rapidly immobilizing the bacteria, thus restraining the spread of infection. It is proposed that this phenomenon may be a significant component of crustacean cellular host defenses.     

Sigma factor is not released during transcription in Bacillus subtilis

Molecular Genetics and Genomics - The relationship between sigma (σ) and delta (δ) factors of Bacillus subtilis RNA polymerase has been analyzed during initiation of RNA synthesis. When...     

Induction by ouabain of hemoglobin synthesis in cultured friend erythroleukemic cells

Induction of erythroid differentiation in ouabain-resistant murine erythroleukemia cells by ouabain is reported. Ouabain induction results in the appearance of hemoglobin-containing cells 12–24 hr earlier than induction of the same clone by dimethyl sulfoxide. The levels of globin mRNA after ouabain induction are similar in amount to the globin mRNA levels observed after induction by dimethyl sulfoxide. The concentration of ouabain required to induce hemoglobin synthesis depends upon the K⁺ ion levels in the culture medium. Lowering the extracellular K⁺ ion concentration 2–4 fold reduced by 10–40 fold the ouabain concentration necessary for the induction of hemoglobin synthesis. In low K⁺ medium (1.8 mM), ouabain is an effective inducer of hemoglobin synthesis at a concentration of 0.02 mM. This K⁺ effect is specific for ouabain induction, since induction by other inducers, such as dimethyl sulfoxide and dimethyl acetamide, does not exhibit this marked sensitivity to the levels of K⁺ ions in the culture medium. These results suggest that the binding of ouabain to the plasma membrane enzyme, $\text{Na}\text{K}$ ATPase, is required for the induction of erythroid differentiation by ouabain. A small but significant proportion of wild-type, ouabain-sensitive cells also can be induced by ouabain, below ouabain concentrations that are toxic to these cells. The observation that the binding of ouabain to the $\text{Na}\text{K}$ ATPase induces hemoglobin synthesis suggests that changes in the intracellular concentration of K⁺ ions may be involved in the control of erythroid differentiation in Friend erythroleukemic cells.     

Ethylene and IAA transport in potato

Although elongation growth is reduced by ethylene, swelling responses do not occur. Ethylene reduces neither transport nor metabolism of applied IAA in either mesocotyl or coleoptile. We propose that maintenance of high auxin levels within these tissues sustains polar transport and contributes to the relative insensitivity of maize to applied ethylene.     

Mice lacking the mitochondrial exonuclease MGME1 develop inflammatory kidney disease with glomerular dysfunction

Mitochondrial DNA (mtDNA) maintenance disorders are caused by mutations in ubiquitously expressed nuclear genes and lead to syndromes with variable disease severity and tissue-specific phenotypes. Loss of function mutations in the gene encoding the mitochondrial genome and maintenance exonuclease 1 (MGME1) result in deletions and depletion of mtDNA leading to adult-onset multisystem mitochondrial disease in humans. To better understand the in vivo function of MGME1 and the associated disease pathophysiology, we characterized a Mgme1 mouse knockout model by extensive phenotyping of ageing knockout animals. We show that loss of MGME1 leads to de novo formation of linear deleted mtDNA fragments that are constantly made and degraded. These findings contradict previous proposal that MGME1 is essential for degradation of linear mtDNA fragments and instead support a model where MGME1 has a critical role in completion of mtDNA replication. We report that Mgme1 knockout mice develop a dramatic phenotype as they age and display progressive weight loss, cataract and retinopathy. Surprisingly, aged animals also develop kidney inflammation, glomerular changes and severe chronic progressive nephropathy, consistent with nephrotic syndrome. These findings link the faulty mtDNA synthesis to severe inflammatory disease and thus show that defective mtDNA replication can trigger an immune response that causes age-associated progressive pathology in the kidney.     

An allied reprogramming,selection, expansion and differentiation platform for creating hiPSC on microcarriers

ObjectivesInduced pluripotent stem cells (iPSCs) generated by monolayer cultures is plagued by low efficiencies, high levels of manipulation and operator unpredictability. We have developed a platform, reprogramming, expansion, and differentiation on Microcarriers, to solve these challenges.Materials and MethodsFive sources of human somatic cells were reprogrammed, selected, expanded and differentiated in microcarriers suspension cultures.ResultsImprovement of transduction efficiencies up to 2 times was observed. Accelerated reprogramming in microcarrier cultures was 7 days faster than monolayer, providing between 30 and 50‐fold more clones to choose from fibroblasts, peripheral blood mononuclear cells, T cells and CD34+ stem cells. This was observed to be due to an earlier induction of genes (β‐catenin, E‐cadherin and EpCAM) on day 4 versus monolayer cultures which occurred on days 14 or later. Following that, faster induction and earlier stabilization of pluripotency genes occurred during the maturation phase of reprogramming. Integrated expansion without trypsinization and efficient differentiation, without embryoid bodies formation, to the three germ‐layers, cardiomyocytes and haematopoietic stem cells were further demonstrated.ConclusionsOur method can solve the inherent problems of conventional monolayer cultures. It is highly efficient, cell dissociation free, can be operated with lower labor, and allows testing of differentiation efficiency without trypsinization and generation of embryoid bodies. It is also amenable to automation for processing more samples in a small footprint, alleviating many challenges of manual monolayer selection.

We have developed an allied protocol for reprogramming, selecting, expanding and differentiating human pluripotent stem cells on Microcarriers (designated as RepMC). This method allows faster reprogramming, selecting 30‐50‐fold more candidates for characterization and also allows us to find high quality candidates that differentiate to cardiomyocytes and blood lineages. Mechanistically, this method appears to accelerate the induction, maturation and stabilization phases of reprogramming. Our findings help simplify the process of deriving and expanding iPSCs for therapeutic applications, offering a robust and scalable suspension platform for large‐scale generation of clinical grade iPSCs.     

Assessing the impacts of fragmentation on plant communities in New Zealand: scaling from survey plots to landscapes

Aim Few studies have attempted to assess the overall impact of fragmentation at the landscape scale. We quantify the impacts of fragmentation on plant diversity by assessing patterns of community composition in relation to a range of fragmentation measures. Location The investigation was undertaken in two regions of New Zealand – a relatively unfragmented area of lowland rain forest in south Westland and a highly fragmented montane forest on the eastern slopes of the Southern Alps. Methods We calculated an index of community similarity (Bray–Curtis) between forest plots we regarded as potentially affected by fragmentation and control forest plots located deep inside continuous forest areas. Using a multiple nonlinear regression technique that incorporates spatial autocorrelation effects, we analysed plant community composition in relation to measures of fragmentation at the patch and landscape levels. From the resulting regression equation, we predicted community composition for every forest pixel on land‐cover maps of the study areas and used these maps to calculate a landscape‐level estimate of compositional change, which we term ‘BioFrag’. BioFrag has a value of one if fragmentation has no detectable effect on communities within a landscape, and tends towards zero if fragmentation has a strong effect. Results We detected a weak, but significant, impact of fragmentation metrics operating at both the patch and landscape levels. Observed values of BioFrag ranged from 0.68 to 0.90, suggesting that patterns of fragmentation have medium to weak impacts on forest plant communities in New Zealand. BioFrag values varied in meaningful ways among landscapes and between the ground‐cover and tree and shrub communities. Main conclusions BioFrag advances methods that describe spatial patterns of forest cover by incorporating the exact spatial patterns of observed species responses to fragmentation operating at multiple spatial scales. BioFrag can be applied to any landscape and ecological community across the globe and represents a significant step towards developing a biologically relevant, landscape‐scale index of habitat fragmentation.