Socio-economic determinants of health and physical fitness in southern Ethiopia

The dependence of health and physical fitness on the socio-economic factors of rural families in southern Ethiopia is investigated, with particular emphasis on the role of inequality. This paper contributes to our knowledge of the effect of inequality on health in several ways: it compares the results of objective and subjective health measures, it distinguishes between wealth inequality and nutrition inequality, and it evaluates the impact of nutrition inequality both at the village level and at the household level. The subjective health measures are the number of days respondents were ill during the last month, their ability to walk distances, their ability to carry heavy loads, and their ability to work in the field. The objective health measure is having Body Mass Index (BMI) lower than 18.5. Males are healthier than females. Height has a positive and significant effect on health and fitness and the same is true for per-capita wealth measured at the village level. Availability of satisfactory health facilities has a negative effect on morbidity. Per-capita wealth inequality is positively associated with morbidity and with a low BMI. Within-household nutrition inequality has a complex effect on health and physical fitness: the effect is negative, but only for household members whose nutritional status is above the household mean. The results indicate a clear positive effect of economic well-being on health and physical fitness. The role of inequality is less clear, and certainly deserves further analyses at both the theoretical and empirical levels.     

Long-term microcarrier suspension cultures of human embryonic stem cells

The conventional method of culturing human embryonic stem cells (hESC) is on two-dimensional (2D) surfaces, which is not amenable for scale up to therapeutic quantities in bioreactors. We have developed a facile and robust method for maintaining undifferentiated hESC in three-dimensional (3D) suspension cultures on matrigel-coated microcarriers achieving 2- to 4-fold higher cell densities than those in 2D colony cultures. Stable, continuous propagation of two hESC lines on microcarriers has been demonstrated in conditioned media for 6 months. Microcarrier cultures (MC) were also demonstrated in two serum-free defined media (StemPro and mTeSR1). MC achieved even higher cell concentrations in suspension spinner flasks, thus opening the prospect of propagation in controlled bioreactors.     

Type 4 pili are dispensable for biofilm development in the cyanobacterium Synechococcus elongatus

The hair‐like cell appendages denoted as type IV pili are crucial for biofilm formation in diverse eubacteria. The protein complex responsible for type IV pilus assembly is homologous with the type II protein secretion complex. In the cyanobacterium Synechococcus elongatus PCC 7942, the gene Synpcc7942_2071 encodes an ATPase homologue of type II/type IV systems. Here, we report that inactivation of Synpcc7942_2071 strongly affected the suite of proteins present in the extracellular milieu (exo‐proteome) and eliminated pili observable by electron microscopy. These results support a role for this gene product in protein secretion as well as in pili formation. As we previously reported, inactivation of Synpcc7942_2071 enables biofilm formation and suppresses the planktonic growth of S. elongatus. Thus, pili are dispensable for biofilm development in this cyanobacterium, in contrast to their biofilm‐promoting function in type IV pili‐producing heterotrophic bacteria. Nevertheless, pili removal is not required for biofilm formation as evident by a piliated mutant of S. elongatus that develops biofilms. We show that adhesion and timing of biofilm development differ between the piliated and non‐piliated strains. The study demonstrates key differences in the process of biofilm formation between cyanobacteria and well‐studied type IV pili‐producing heterotrophic bacteria.     

Versatility of EGF receptor ligand processing in insects

Processing of EGF-family ligands is an essential step in triggering the EGF receptor pathway, which fulfills a diverse set of roles during development and tissue maintenance. We describe a mechanism of ligand processing which is unique to insects, and possibly to other invertebrates. This mechanism relies on ligand precursor trafficking from the ER by a chaperone, Star (S), and precursor cleavage by Rhomboids, a family of intra-membrane protease. Remarkably, the ability of Rhomboids to cleave S as well, endows the pathway with additional diversity. Rhomboid isoforms which also reside in the ER inactivate the chaperone before any ligand was trafficked, thus significantly reducing the level of ligand that will eventually be processed and secreted. ER localization also serves as a critical feature in trafficking the entire ligand-processing machinery to axonal termini, as the ER extends throughout the axon. Finally, examination of diverse species of insects demonstrates the evolution of chaperone cleavability, indicating that the primordial processing machinery could support long-range signaling by the ligand. Altering the intracellular localization of critical components of a conserved signaling cassette therefore provides an evolutionary mechanism for modulation of signaling levels, and diversification of the biological settings where the pathway functions.     

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.     

Cholesterol trafficking and raft-like membrane domain composition mediate scavenger receptor class B type 1-dependent lipid sensing in intestinal epithelial cells

Scavenger receptor Class B type 1 (SR-B1) is a lipid transporter and sensor. In intestinal epithelial cells, SR-B1-dependent lipid sensing is associated with SR-B1 recruitment in raft-like/ detergent-resistant membrane domains and interaction of its C-terminal transmembrane domain with plasma membrane cholesterol. To clarify the initiating events occurring during lipid sensing by SR-B1, we analyzed cholesterol trafficking and raft-like domain composition in intestinal epithelial cells expressing wild-type SR-B1 or the mutated form SR-B1-Q445A, defective in membrane cholesterol binding and signal initiation. These features of SR-B1 were found to influence both apical cholesterol efflux and intracellular cholesterol trafficking from plasma membrane to lipid droplets, and the lipid composition of raft-like domains. Lipidomic analysis revealed likely participation of d18:0/16:0 sphingomyelin and 16:0/0:0 lysophosphatidylethanolamine in lipid sensing by SR-B1. Proteomic analysis identified proteins, whose abundance changed in raft-like domains during lipid sensing, and these included molecules linked to lipid raft dynamics and signal transduction. These findings provide new insights into the role of SR-B1 in cellular cholesterol homeostasis and suggest molecular links between SR-B1-dependent lipid sensing and cell cholesterol and lipid droplet dynamics.