Amaranth seed oil was fractionated in a bench‐scale short‐path distillation unit to obtain fractions rich in squalene. Fractionations were conducted with degummed amaranth oil, alkali‐refined amaranth oil, and simulated amaranth oil. Squalene concentration was increased about sevenfold with a squalene recovery of 76.0% in the distillate when degummed amaranth oil was fractionated at 180°C and 3 mtorr vacuum. Free fatty acids codistilled with squalene, lowering the squalene content of the distillate, and resulted in a semisolid distillate at room temperature. Alkali‐refining was subsequently used to reduce the free fatty acid content before fractionation. A simulated oil (7% squalene/93% soybean oil) and alkali‐refined amaranth oil were fractionated at three temperatures (160, 170, and 180°C) and five vacuum settings (10, 100, 200, 400, and 600 mtorr). The highest squalene recoveries from simulated oil and alkali‐refined amaranth oil were 73.4 and 67.8%, respectively, both at 180°C and 100 mtorr, which translates to 12.1‐and 9.2‐fold increases in squalene concentration, respectively. The squalene recovery of the alkali‐refined amaranth oil at 180°C was not significantly different at 10 mtorr vs. 100 mtorr. The results of this study can be used as a component to assess the economic feasibility of fractionating amaranth seed for starch, oil, meal, and squalene.

No abstract

Microglia are a specialized population of myeloid cells that mediate CNS innate immune responses. Efforts to identify the cellular and molecular mechanisms that regulate microglia behaviors have been hampered by the lack of effective tools for manipulating gene expression. Cultured microglia are refractory to most chemical and electrical transfection methods, yielding little or no gene delivery and causing toxicity and/or inflammatory activation. Recombinant adeno-associated viral (rAAVs) vectors are non-enveloped, single-stranded DNA vectors commonly used to transduce many primary cell types and tissues. In this study, we evaluated the feasibility and efficiency of utilizing rAAV serotype 2 (rAAV2) to modulate gene expression in cultured microglia. rAAV2 yields high transduction and causes minimal toxicity or inflammatory response in both neonatal and adult microglia. To demonstrate that rAAV transduction can induce functional protein expression, we used rAAV2 expressing Cre-recombinase to successfully excise a LoxP-flanked miR155 gene in cultured microglia. We further evaluated rAAV serotypes 5, 6, 8, and 9, and observed that all efficiently transduced cultured microglia to varying degrees of success and caused little or no alteration in inflammatory gene expression. These results provide strong encouragement for the application of rAAV-mediated gene expression in microglia for mechanistic and therapeutic purposes.

No abstract

Fusarium graminearum, a known producer of trichothecene mycotoxins in cereal hosts, has been recently documented as a cause of dry rot of potato tubers in the United States. Due to the uncertainty of trichothecene production in these tubers, a study was conducted to determine the accumulation and diffusion of trichothecenes in potato tubers affected with dry rot caused by F. graminearum. Potato tubers of cv. Russet Burbank were inoculated with 14 F. graminearum isolates from potato, sugar beet, and wheat and incubated at 10 to 12 degrees C for 5 weeks to determine accumulation of trichothecenes in potato tubers during storage. Twelve of the isolates were classified as deoxynivalenol (DON) genotype and two isolates were as nivalenol (NIV) genotype. Trichothecenes were detected only in rotted tissue. DON was detected in all F. graminearum DON genotype isolates up to 39.68 microg/ml in rotted potato tissue. Similarly, both NIV genotype isolates accumulated NIV in rotted potato tissue up to 18.28 microg/ml. Interestingly, isolates classified as genotype DON accumulated both DON and NIV in the dry rot lesion. Potato tubers were then inoculated with two isolates of F. graminearum chemotype DON and incubated up to 7 weeks at 10 to 12 degrees C and assayed for DON diffusion. F. graminearum was recovered from >53% of the isolations from inoculated tubers at 3 cm distal to the rotted tissue after 7 weeks of incubation but DON was not detected in the surrounding tissue. Based in this data, the accumulation of trichothecenes in the asymptomatic tissue surrounding dry rot lesions caused by F. graminearum is minimal in cv. Russet Burbank potato tubers stored for 7 weeks at customary processing storage temperatures.