The quantitation of sugars, including glucose, the primary fermentable sugar; maltose (DP2); and maltotriose (DP3), is a standard procedure during the corn-to-ethanol fermentation process. The quantitation of glucose by the Megazyme Assay utilizing glucose oxidase and peroxidase enzymes (GOPOD) and UV-Vis detection, high-performance liquid chromatography with refractive index detection (HPLC-RID), and liquid chromatography mass spectrometry (LC-MS) with electrospray ionization (ESI) and selected ion monitoring (SIM) was studied. Three biological flask fermentation replicates were analyzed every 12 h beginning at 14 h of fermentation (T14) until near completion of fermentation (T62). The method comparison results for glucose quantitation showed that the LC-MS SIM analysis had the lowest limit of quantitation (LOQ) at 2 ppm and the widest dynamic range of 2.7 orders of magnitude. The HPLC-RID analysis had a linear dynamic range (LDR) of 1.5 orders of magnitude with an LOQ of 1500 ppm. The Megazyme GOPOD analysis had an LDR of 0.9 orders of magnitude with an LOQ of 120 ppm. The HPLC-RID method was ideal for glucose quantitation when it was present in high concentrations. In contrast, maltose and maltotriose components were found to be present in lower concentrations, such that simultaneous quantitation of the three analytes is difficult during fermentation. The LC-MS method was the only method able to quantify the concentration of glucose successfully and simultaneously with DP2 and DP3 in all the fermentation broth samples collected from T14 through T62 during the corn-to-ethanol fermentation process.
Background and Objectives Antibiotics are used in the corn‐based ethanol production to control the proliferation of bacteria during fermentation of starch to ethanol. Dried distiller's grains with solubles (DDGS) is a by‐product of corn‐based fermentation that can effectively be used as feed for cattle, swine, chicken, and other livestock. High amounts of antibiotic residues in the feed may lead to antibiotic resistant bacteria. Findings Antibiotics were extracted from DDGS samples by liquid‐liquid extraction, purified by solid phase extraction, and quantified by liquid chromatography mass spectrometry in multiple reaction monitoring mode to quantify five antibiotics: penicillin G, erythromycin, tylosin, virginiamycin M1, and virginiamycin S1. DDGS was collected and analyzed quarterly over the course of ten months to obtain a representation of DDGS produced in various parts of the United States in different seasons. Conclusions Penicillin G was detected in a single sample. Tylosin was detected in 28%, erythromycin in 45%, virginiamycin M1 in 53%, and virginiamycin S1 in 83% of the total samples. Significance and Novelty Largest study to date to quantify antibiotics in DDGS (33 ethanol plants in 14 states).
The primary purpose of this study was to determine prevalence of the Male Athlete Triad (MAT) conditions: low energy availability (EA), low bone mineral density (BMD), and low testosterone in male collegiate athletes from different sports. Participants included 44 collegiate male athletes (age, 20.4 ± 0.2 yr; BMI, 25.3 ± 1.3 kg/m2) from seven sports (cross country, soccer, basketball, wrestling, track, golf, and baseball). Resting metabolic rate, three-day food intake, seven-day exercise energy expenditure, body composition, and reproductive and metabolic hormones were assessed. Of the total participants, 15% had low EA, 0% had low BMD, 28% had low total testosterone (TT), and 80% had low calculated free testosterone (cFT). There were no significant correlations between EA, BMD, TT, and cFT. Insulin and sex hormone binding globulin (SHBG) were below and on the upper end of the reference range for healthy male adults, respectively. Insulin was negatively correlated with total (r = -0.330, p = 0.043) and lumbar spine BMD z-scores (r = -0.413, p = 0.010). Low TT and low cFT were the most prevalent MAT conditions among all athletes. Further research should investigate the relationship between insulin and SHBG and the role of these hormones in the MAT. Novelty Bullets • Assessment of energy availability alone is not sufficient to identify physiological disturbances in collegiate male athletes. • Low total and/or free testosterone may be present in some collegiate male athletes, regardless of BMD status. • Low insulin and high SHBG concentration may portray the presence of conditions of the MAT in male collegiate athletes.
Aim: Sex differentiation occurs during the perinatal period (four days before and after birth) in the rat brain. The brain remains a female brain unless exposed to testosterone, which gets metabolized to 17β-estradiol and dihydrotestosterone. While aromatase converts testosterone to 17β-estradiol, 5α-reductase converts testosterone to dihydrotestosterone. 17β-estradiol exerts its effects by binding to an estrogen receptor, whereas dihydrotestosterone via an androgen receptor. The role of dihydrotestosterone and 17β-estradiol during the organizational phase of brain differentiation was tested using adult rat behaviors. The study sought to examine the effects of perinatal manipulations on adult rat behaviors. Materials and methods: Timed-pregnant rats received either the vehicle (5% ethanol and 95% sesame oil), exemestane (4 mg/kg/ml; aromatase inhibitor), or flutamide (20 mg/kg/ml; androgen receptor blocker) subcutaneously during the last four days of pregnancy. Pups from each group continued to receive their treatments during the first four days after birth. From postnatal day 65, these animals were subjected to open field, sexual motivation, and spatial working memory tests. Results: Male rats receiving flutamide exhibited exploratory behavior significantly more compared to exemestane and control groups. Male rats receiving exemestane had substantially higher plasma testosterone and displayed an increased interest in the estrus rat than the control and flutamide group. Conclusion: Androgen receptor blockade resulted in increased exploratory behavior in male rats where 17β-estradiol was testosterone's primary metabolite. The blocking of aromatase using exemestane resulted in higher levels of plasma testosterone and enhanced sexual motivation in male rats. Thus, 17β-estradiol and dihydrotestosterone's presence during the organizational phase is essential for normal socio-sexual and exploratory behaviors.
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