Significance of agitation-induced shear stress on mycelium morphology and lavendamycin production by engineered Streptomyces flocculus

Lavendamycin methyl ester (LME) is a derivative of a highly functionalized aminoquinone alkaloid lavendamycin and could be used as a scaffold for novel anticancer agent development. This work demonstrated LME production by cultivation of an engineered strain of Streptomyces flocculus CGMCC4.1223 ΔstnB1, while the wild-type strain did not produce. To enhance its production, the effect of shear stress and oxygen supply on ΔstnB1 strain cultivation was investigated in detail. In flask culture, when the shaking speed increased from 150 to 220 rpm, the mycelium was altered from a large pellet to a filamentous hypha, and the LME production was almost doubled, while no significant differences were observed among varied filling volumes, which implied a crucial role of shear stress in the morphology and LME production. To confirm this suggestion, experiments with agitation speed ranging from 400 to 1,000 rpm at a fixed aeration rate of 1.0 vvm were conducted in a stirred tank bioreactor. It was found that the morphology became more hairy with reduced pellet size, and the LME production was enhanced threefolds when the agitation speed increased from 400 to 800 rpm. Further experiments by varying initial k _L a value at the same agitation speed indicated that oxygen supply only slightly affected the physiological status of ΔstnB1 strain. Altogether, shear stress was identified as a major factor affecting the cell morphology and LME production. The work would be helpful to the production of LME and other secondary metabolites by filamentous microorganism cultivation.     

Olfactory Cues Used in Host Selection by Frankliniella occidentalis (Thysanoptera: Thripidae) in Relation to Host Suitability

Olfactory responses of Frankliniella occidentalis (Pergande) to odors from six vegetable host plants-cabbage (Brassica oleracea), lettuce (Lactuca sativa), cucumber (Cucumis sativus), eggplant (Solanum melongena), celery (Apium graveolens) and garlic (Allium sativum) - were measured in choice tests in a Y-tube olfactometer, while host suitability of these same plants was assessed based on life table parameters-development time, survival rates, fertility, offspring sex ratio, female longevity, and an index of population change (I) - measured on leaf disks. Olfactory response was measured on both undamaged and mechanically-damaged plants. Regardless of plant damaged, female F. occidentalis responded positively to the odors of cabbage, lettuce, cucumber, and eggplant, but showed negative responses to celery and garlic, when offered a choice of plant odor or clean air. When female thrips were offered choices between each possible pairing of undamaged plants, or pairings of mechanically-damaged plants, the order of preference was cabbage = lettuce = cucumber > eggplant > celery = garlic. Male thrips responded differently, males responded positively only to undamaged lettuce and eggplant compared to clean air, while among mechanically-damaged plants only cucumber elicited a positive response. Given a choice between all possible pairings of undamaged plants, male thrips preferred lettuce to celery, and eggplant to garlic, whereas among pairings of mechanically-damaged plants male thrips preferred lettuce to celery, and cucumber to eggplant. The order of suitability of the six host plants based on life table parameters for F. occidentalis was essentially the same as the order of preference by female thrips. These results indicate that host plant odor is an important cue for female F. occidentalis in recognizing hosts suitable for oviposition. Information on plant attraction and repellency may be useful in thrips pest management. In addition, the correspondence of preference and performance that we found suggests that evolution of local host adaptation and, perhaps, host specialization, is possible.     

Interferon-γ Induces Immunoproteasomes and the Presentation of MHC I-Associated Peptides on Human Salivary Gland Cells

A prominent histopathological feature of Sjögren''s syndrome, an autoimmune disease, is the presence of lymphocytic infiltrates in the salivary and lachrymal glands. Such infiltrates are comprised of activated lymphocytes and macrophages, and known to produce multiple cytokines including interferon-gamma (IFN-γ). In this study, we have demonstrated that IFN-γ strongly induces the expression of immunoproteasome beta subunits (β1i, β2i and β5i) and immunoproteasome activity but conversely inhibits the expression of proteasome beta subunits (β1, β2 and β5) in human salivary gland (HSG) cells. Mass spectrometric analysis has revealed potential MHC I-associated peptides on the HSG cells, including a tryptic peptide derived from salivary amylase, due to IFN-γ stimulation. These results suggest that IFN-γ induces immunoproteasomes in HSG cells, leading to enhanced presentation of MHC I-associated peptides on cell surface. These peptide-presenting salivary gland cells may be recognized and targeted by auto-reactive T lymphocytes. We have also found that lactacystin, a proteasome inhibitor, inhibits the expression of β1 subunit in HSG cells and blocks the IFN-γ-induced expression of β1i and immunoproteasome activity. However, the expression of β2i and β5i in HSG cells is not affected by lactacystin. These results may add new insight into the mechanism regarding how lactacystin blocks the action of proteasomes or immunoproteasomes.     

Contribution of the skin,rectus abdominis and their sheaths to the structural response of the abdominal wall ex vivo

A better understanding of the abdominal wall biomechanics could help designing new treatments for incisional hernia. In the current study, an experimental protocol was developed to evaluate the contributions of the abdominal wall components to the structural response of the anterior part of the abdominal wall. The specimens underwent 3 dissections (removal of (1) skin and subcutaneous fat, (2) anterior rectus sheath, (3) rectus abdominis muscles). After each dissection, they were subjected to air pressure up to 3 kPa. Ultrasound images and associated elastographic maps were collected at 0, 2 and 3 kPa in the intact state and strains on the internal surface were calculated using stereo-correlation in all states. Strains on the rectus abdominis and linea alba were analyzed. After the dissection of the anterior sheath of the rectus abdominis, longitudinal strain was found significantly different on the linea alba (5% at 3 kPa) and on the rectus abdominis area (11% at 3 kPa). The current results highlight the importance of the rectus sheath in the structural response of the anterior part of the abdominal wall ex vivo. Geometrical characteristics such as thicknesses and radii of curvature and mechanical properties (shear modulus of the rectus abdominis, e.g. at 0 pressure the average value is 14 kPa) were provided in order to facilitate future modeling efforts.     

Pain Reduction and Financial Incentives to Improve Glucose Monitoring Adherence in a Community Health Center

Self-monitoring of blood glucose is a critical component of diabetes management. However, patients often do not maintain the testing schedule recommended by their healthcare provider. Many barriers to testing have been cited, including cost and pain. We present a small pilot study to explore whether the use of financial incentives and pain-free lancets could improve adherence to glucose testing in a community health center patient population consisting largely of non-English speaking ethnic minorities with low health literacy. The proportion of patients lost to follow-up was 17%, suggesting that a larger scale study is feasible in this type of setting, but we found no preliminary evidence suggesting a positive effect on adherence by either financial incentives or pain-free lancets. Results from this pilot study will guide the design of larger-scale studies to evaluate approaches to overcome the variety of barriers to glucose testing that are present in disadvantaged patient populations.