Three-dimensional microscale hanging drop arrays with geometric control for drug screening and live tissue imaging.

Existing three-dimensional (3D) culture techniques are limited by trade-offs between throughput, capacity for high-resolution imaging in living state, and geometric control. Here, we introduce a modular microscale hanging drop culture where simple design elements allow high replicates for drug screening, direct on-chip real-time or high-resolution confocal microscopy, and geometric control in 3D. Thousands of spheroids can be formed on our microchip in a single step and without any selective pressure from specific matrices. Microchip cultures from human LN229 glioblastoma and patient-derived mouse xenograft cells retained genomic alterations of originating tumors based on mate pair sequencing. We measured response to drugs over time with real-time microscopy on-chip. Last, by engineering droplets to form predetermined geometric shapes, we were able to manipulate the geometry of cultured cell masses. These outcomes can enable broad applications in advancing personalized medicine for cancer and drug discovery, tissue engineering, and stem cell research.

Effects of crystalline amino acid supplementation to the diet on odor from pig manure.

The objective of this study was to determine the effects of specific crystalline AA supplementation to a diet on odor emission, odor intensity, odor hedonic tone, and ammonia emission from pig manure, and on manure characteristics (pH; ammonia N; total nitrogen; sulfurous, indolic, and phenolic compounds; and VFA concentrations). An experiment was conducted with growing pigs (n = 18) in a randomized complete block design, with 3 treatments in 6 blocks. Treatment groups were (1) a 15%-CP basal diet with 3 times the requirement of sulfur-containing AA (14.2 g/kg of diet, as-fed basis); (2) the basal diet with 2 times the requirement of Trp and Phe+Tyr (2.9 and 20.4 g/kg of diet, respectively, as-fed basis); and (3) the basal diet with AA supplementation to levels sufficient for maximum protein gain. Pigs with an initial BW of 41.2 +/- 0.8 kg were individually penned in partly slatted floor pens and offered a daily feed allowance of 2.8 times the maintenance requirement for NE (293 kJ/kg of BW(0.75)). Feed was mixed with water at 1:2.5 (wt/wt). Feces and urine of each pig was allowed to accumulate in separate manure pits under the slatted floor. After an adaptation period of 2 wk, and after cleaning the manure pits, manure was subsequently collected. In wk 5 of the collection period, separate samples were collected directly from each manure pit for odor, ammonia, and manure composition analyses. Air samples were analyzed for odor concentration and for hedonic tone and odor intensity above the odor detection threshold. Results showed that supplementing crystalline S-containing AA in surplus of the requirement increased odor emission (P < 0.001) and odor intensity (P < 0.05) and reduced odor hedonic tone (P < 0.05) from the air above the manure pits. Supplementing crystalline Trp, Tyr, and Phe in surplus of the recommended requirements did not affect odor emission, odor intensity, or odor hedonic tone. Regardless of dietary treatment, all pigs had similar performance levels. No differences were observed in ammonia emission from manure of pigs fed different levels of AA supplementation (P = 0.20). To reduce odor from pig manure, dietary S-containing AA should be minimized to just meet the recommended requirements.

Characteristics of Pseudomonas aeruginosa isolated from whirlpools and bathers.

Pseudomonas aeruginosa is the most frequently isolated microorganism from whirlpool water and lesions associated with outbreaks of dermatitis and folliculitis related to whirlpool exposure. Strains were selected from 19 outbreaks of P. aeruginosa infections (1977 to 1983) associated with whirlpool use; they were examined to determine if the strains possessed unique virulence factors or characteristics that might aid in their selection in the environment. P. aeruginosa, 011, was the predominant serotype isolated from whirlpool water as well as from bathers with dermatitis or folliculitis, followed by serotypes 09, 04, and 03. Antimicrobial susceptibility patterns were similar for all strains. Strains of P. aeruginosa from bathers and water demonstrated statistically significant differences in extracellular enzyme production compared with control strains. P. aeruginosa, serotypes 09 and 011, were found to be sensitive to low levels of chlorine. These data suggest that, if adequate levels of free available chlorine are maintained, P. aeruginosa should have little opportunity to persist in whirlpools. A bather's risk of P. aeruginosa dermatitis or folliculitis appears to be affected primarily by three factors: immersion in water colonized by P. aeruginosa, skin hydration with altered skin flora, and toxic reactions to extracellular enzyme or exotoxins produced by P. aeruginosa. Although a single virulence factor was not identified from the results of this study, there are some indications that the enzymes produced by these microorganisms play an important role in the pathogenesis of disease associated with whirlpool use.