Mutations in the Lipopolysaccharide Biosynthesis Pathway Interfere with Crescentin-Mediated Cell Curvature in Caulobacter crescentus

Bacterial cell morphogenesis requires coordination among multiple cellular systems, including the bacterial cytoskeleton and the cell wall. In the vibrioid bacterium Caulobacter crescentus, the intermediate filament-like protein crescentin forms a cell envelope-associated cytoskeletal structure that controls cell wall growth to generate cell curvature. We undertook a genetic screen to find other cellular components important for cell curvature. Here we report that deletion of a gene (wbqL) involved in the lipopolysaccharide (LPS) biosynthesis pathway abolishes cell curvature. Loss of WbqL function leads to the accumulation of an aberrant Opolysaccharide species and to the release of the S layer in the culture medium. Epistasis and microscopy experiments show that neither S-layer nor O-polysaccharide production is required for curved cell morphology per se but that production of the altered O-polysaccharide species abolishes cell curvature by apparently interfering with the ability of the crescentin structure to associate with the cell envelope. Our data suggest that perturbations in a cellular pathway that is itself fully dispensable for cell curvature can cause a disruption of cell morphogenesis, highlighting the delicate harmony among unrelated cellular systems. Using the wbqL mutant, we also show that the normal assembly and growth properties of the crescentin structure are independent of its association with the cell envelope. However, this envelope association is important for facilitating the local disruption of the stable crescentin structure at the division site during cytokinesis

The effect of scale-free topology on the robustness and evolvability of genetic regulatory networks

We investigate how scale-free (SF) and Erdos-Renyi (ER) topologies affect the interplay between evolvability and robustness of model gene regulatory networks with Boolean threshold dynamics. In agreement with Oikonomou and Cluzel (2006) we find that networks with SFin topologies, that is SF topology for incoming nodes and ER topology for outgoing nodes, are significantly more evolvable towards specific oscillatory targets than networks with ER topology for both incoming and outgoing nodes. Similar results are found for networks with SFboth and SFout topologies. The functionality of the SFout topology, which most closely resembles the structure of biological gene networks (Babu et al., 2004), is compared to the ER topology in further detail through an extension to multiple target outputs, with either an oscillatory or a non-oscillatory nature. For multiple oscillatory targets of the same length, the differences between SFout and ER networks are enhanced, but for non-oscillatory targets both types of networks show fairly similar evolvability. We find that SF networks generate oscillations much more easily than ER networks do, and this may explain why SF networks are more evolvable than ER networks are for oscillatory phenotypes. In spite of their greater evolvability, we find that networks with SFout topologies are also more robust to mutations than ER networks. Furthermore, the SFout topologies are more robust to changes in initial conditions (environmental robustness). For both topologies, we find that once a population of networks has reached the target state, further neutral evolution can lead to an increase in both the mutational robustness and the environmental robustness to changes in initial conditions.Comment: 16 pages, 15 figure

The Effect of Acute Consumption of Overtime Essential Amino Acids Sports Drink on Delayed Onset Muscle Soreness in the Older Sedentary Population

Essential amino acids are necessary for nutrition whether obtained by digestion of proteins or by oral supplementation of amino acids. The elderly experience loss of skeletal muscles and decrease in their strength and function, which can lead to poor quality of life. Increased quantity and quality of proteins stimulates muscle protein synthesis that can help combat this natural aging process. Purpose: The aim of this study was to see if consumption of essential amino acid drink high concentrations of leucin (2040 mg/serving) during exercise will attenuate the condition of Delayed Onset Muscle Soreness in the elderly. Also, we aimed to assess the degree of muscle flexibility and endurance followed the three-day exercise protocol. We hypothesized that the older participants acutely ingesting the essential amino acid supplement during the exercise regime will have increased physical performance and diminished symptoms of DOMS. Methods: In this study, 16 participants (6 P, 10 EAA-O) completed a health screening visit and an exercise routine (sit, stretch and reach, shoulder flexibility distance, MVC isometric handgrip, push-ups (reps), flexed arm hang (time), cable triceps extension (50% of one repetition max until failure), and a 1.5-mile run, with intermittent consumption of the sports drink) for three consecutive days. The study participants were randomly assigned to either the EAA-O group (6.6g of EAA-O + Gatorade) or the control group (Gatorade-only). The study design is a double blinded study as neither the recording analysis researchers nor study participants were aware of the assigned group. Results: The EAA-O group improved significantly from day one to two in flexed arm hang (p = 0.036) and the 1.5-mile run (p = 0.040). The EAA-O group improved significantly from day two to three in push-ups (p = 0.002), flexed arm hang (p = 0.035), and 1.5-mile run (0.001). The EAA-O group improved significantly from day one to three in push-ups (p = 0.045), flexed arm hang (p = 0.006), 1.5-mile run (p = 0.0003), and the top speed (p = 0.026). The placebo group did not improve significantly in any of the exercise parameters. Conclusions: Results suggest that the Overtime essential amino acid supplementation combined with training improves overall athletic performance in the older sedentary population. Research reported in this publication was supported by a research contract with Calwood Nutritionals and was approved by the ENMU IRB

Research poster: Water source partitioning for shrubland transpiration using innovative field methods

Research poste

Canalization in the Critical States of Highly Connected Networks of Competing Boolean Nodes

Canalization is a classic concept in Developmental Biology that is thought to be an important feature of evolving systems. In a Boolean network it is a form of network robustness in which a subset of the input signals control the behavior of a node regardless of the remaining input. It has been shown that Boolean networks can become canalized if they evolve through a frustrated competition between nodes. This was demonstrated for large networks in which each node had K=3 inputs. Those networks evolve to a critical steady-state at the boarder of two phases of dynamical behavior. Moreover, the evolution of these networks was shown to be associated with the symmetry of the evolutionary dynamics. We extend these results to the more highly connected K>3 cases and show that similar canalized critical steady states emerge with the same associated dynamical symmetry, but only if the evolutionary dynamics is biased toward homogeneous Boolean functions.Comment: 8 pages, 5 figure

Mineral deposition and vascular invasion of hydroxyapatite reinforced collagen scaffolds seeded with human adipose-derived stem cells

Background: Collagen-based scaffolds reinforced with hydroxyapatite (HA) are an attractive choice for bone tissue engineering because their composition mimics that of bone. We previously reported the development of compression-molded collagen-HA scaffolds that exhibited high porosity, interconnected pores, and mechanical properties that were well-suited for surgical handling and fixation. The objective of this study was to investigate these novel collagen-HA scaffolds in combination with human adipose-derived stem cells (hASCs) as a template for bone formation in a subcutaneous athymic mouse model. Methods: Collagen-HA scaffolds and collagen-only scaffolds were fabricated as previously described, and a clinically approved bone void filler was used as a control for the material. Constructs were seeded with hASCs and were pre-treated with either control or osteogenic media. A cell-free group was also included. Scaffolds were implanted subcutaneously in the backs of athymic nude mice for 8 weeks. Mineral deposition was quantified via micro-computed tomography. Histological and immunofluorescence images of the explants were used to analyze their vascular invasion, remodeling and cellularity. Results: Cell-free collagen-HA scaffolds and those that were pre-seeded with osteogenically differentiated hASCs supported mineral deposition and vascular invasion at comparable rates, while cell-seeded constructs treated with the control medium showed lower mineralization after implantation. HA-reinforcement allowed collagen constructs to maintain their shape, provided improved cell-tissue-scaffold integration, and resulted in a more organized tissue when pre-treated in an osteogenic medium. Scaffold type and pre-treatment also determined osteoclast activity and therefore potential remodeling of the constructs. Conclusions: The results of this study cumulatively indicate that treatment medium and scaffold composition direct mineralization and angiogenic tissue formation in an ectopic model. The data suggest that it may be necessary to match the scaffold with a particular cell type and cell-specific pre-treatment to achieve optimal bone formation

Performance of PCA3 and TMPRSS2:ERG urinary biomarkers in prediction of biopsy outcome in the Canary Prostate Active Surveillance Study (PASS).

BackgroundFor men on active surveillance for prostate cancer, biomarkers may improve prediction of reclassification to higher grade or volume cancer. This study examined the association of urinary PCA3 and TMPRSS2:ERG (T2:ERG) with biopsy-based reclassification.MethodsUrine was collected at baseline, 6, 12, and 24 months in the multi-institutional Canary Prostate Active Surveillance Study (PASS), and PCA3 and T2:ERG levels were quantitated. Reclassification was an increase in Gleason score or ratio of biopsy cores with cancer to ≥34%. The association of biomarker scores, adjusted for common clinical variables, with short- and long-term reclassification was evaluated. Discriminatory capacity of models with clinical variables alone or with biomarkers was assessed using receiver operating characteristic (ROC) curves and decision curve analysis (DCA).ResultsSeven hundred and eighty-two men contributed 2069 urine specimens. After adjusting for PSA, prostate size, and ratio of biopsy cores with cancer, PCA3 but not T2:ERG was associated with short-term reclassification at the first surveillance biopsy (OR = 1.3; 95% CI 1.0-1.7, p = 0.02). The addition of PCA3 to a model with clinical variables improved area under the curve from 0.743 to 0.753 and increased net benefit minimally. After adjusting for clinical variables, neither marker nor marker kinetics was associated with time to reclassification in subsequent biopsies.ConclusionsPCA3 but not T2:ERG was associated with cancer reclassification in the first surveillance biopsy but has negligible improvement over clinical variables alone in ROC or DCA analyses. Neither marker was associated with reclassification in subsequent biopsies