Plasmin in Milk: Activity Measurement, Effect of Environmental Factors, and Correlation with Milk Coagulation

Bovine plasmin activity was measured on H-D-valyl-L-leucyl-L-lysyl-4-nitroanilide by following absorbance changes at 405 nm. Steady-state kinetic parameters Vmax, Km, KI, and KI\u27 were estimated. Bovine plasmin is competitively inhibited by casein and has a Kcat of .0158 ΔA405/min/nM, Km of .107 mM substrate, and KI of .86 mg/ml casein. Bovine plasmin can be measured directly in bovine milk without interference from casein. A total of 380 milk samples from nineteen Holstein (one herd) and nineteen Jersey (one herd) cows was collected monthly during one lactation period. Samples from each cow were analyzed for fat, protein, plasmin activity, plasminogen, pH, SCC, clotting time, curd firming rate, and final curd firmness. Three age groups form each breed/herd were chosen; first, third, and fourth and later lactations. Plasmin activity in milk was most affected by lactation number, with milk from fourth- and later-lactation cows having higher activity than milk from first- or third- lactation cows. Plasmin activity in milk increased during lactation but was not affected by breed/herd, pH, protein, or fat. Plasminogen averaged 5.4 times the plasmin activity in milk and increased during the first five months of lactation. Plasmin activity was higher in milk collected ruing summer and fall but plasminogen was higher in milk collected during fall and winter. Percentage of the total (plasmin+plasminogen) enzyme activated to plasmin increased in late-lactation milk and in milk from fourth- and later-lactation cows. Plasmin activity did not affect any milk clotting parameters in this study. Increased protein in milk resulted in shorter clotting times. When statistically adjusted for protein content, clotting time was longer in milk from the Holstein herd compared to the Jersey herd. Curd firming rate was increased in milk with higher protein and fat. Milk samples collected in the fall had faster firming rates than milk from other seasons. Firming rates remained constant during lactation but increased with higher protein and fat content. Jersey herd milk produced firmer curd than Holstein herd milk and milk collected in the fall had firmer curd than during the other seasons

A whey-protein supplement increases fat loss and spares lean muscle in obese subjects: a randomized human clinical study

<p>Abstract</p> <p>Background</p> <p>This study evaluated a specialized whey fraction (Prolibra™, high in leucine, bioactive peptides and milk calcium) for use as a dietary supplement to enhance weight loss.</p> <p>Methods</p> <p>This was a randomized, double-blind, parallel-arm, 12-week study. Caloric intake was reduced 500 calories per day. Subjects consumed Prolibra or an isocaloric ready-to-mix beverage 20 minutes before breakfast and 20 minutes before dinner. Body fat and lean muscle tissue were measured by dual-energy x-ray absorptiometry (DEXA). Body weight and anthropometric measurements were recorded every 4 weeks. Blood samples were taken at the beginning and end of the study. Statistical analyses were performed on all subjects that completed (completer analysis) and all subjects that lost at least 2.25 kg of body weight (responder analysis). Within group significance was determined at <it>P </it>< 0.05 using a two-tailed paired t-test and between group significance was determined using one way analysis of covariance with baseline data as a covariate.</p> <p>Results</p> <p>Both groups lost a significant amount of weight and the Prolibra group tended to lose more weight than the control group; however the amount of weight loss was not significantly different between groups after 12 weeks. Prolibra subjects lost significantly more body fat compared to control subjects for both the completer (2.81 vs. 1.62 kg <it>P </it>= 0.03) and responder (3.63 vs. 2.11 kg, <it>P </it>= 0.01) groups. Prolibra subjects lost significantly less lean muscle mass in the responder group (1.07 vs. 2.41 kg, <it>P </it>= 0.02). The ratio of fat to lean loss (kg fat lost/kg lean lost) was much larger for Prolibra subjects for both completer (3.75 vs. 1.05) and responder (3.39 vs. 0.88) groups.</p> <p>Conclusion</p> <p>Subjects in both the control and treatment group lost a significant amount of weight with a 500 calorie reduced diet. Subjects taking Prolibra lost significantly more body fat and showed a greater preservation of lean muscle compared to subjects consuming the control beverage. Because subjects taking Prolibra lost 6.1% of their body fat mass, and because a 5% reduction of body fat mass has been shown to reduce the risk of obesity related disease, the results have practical significance.</p

Structure-guided engineering of Lactococcus lactis alcohol dehydrogenase LlAdhA for improved conversion of isobutyraldehyde to isobutanol

We have determined the X-ray crystal structures of the NADH-dependent alcohol dehydrogenase LlAdhA from Lactococcus lactis and its laboratory-evolved variant LlAdhA^(RE1) at 1.9 Å and 2.5 Å resolution, respectively. LlAdhA^(RE1), which contains three amino acid mutations (Y50F, I212T, and L264V), was engineered to increase the microbial production of isobutanol (2-methylpropan-1-ol) from isobutyraldehyde (2-methylpropanal). Structural comparison of LlAdhA and LlAdhA^(RE1) indicates that the enhanced activity on isobutyraldehyde stems from increases in the protein's active site size, hydrophobicity, and substrate access. Further structure-guided mutagenesis generated a quadruple mutant (Y50F/N110S/I212T/L264V), whose K_M for isobutyraldehyde is ∼17-fold lower and catalytic efficiency (k_(cat)/K_M) is ∼160-fold higher than wild-type LlAdhA. Combining detailed structural information and directed evolution, we have achieved significant improvements in non-native alcohol dehydrogenase activity that will facilitate the production of next-generation fuels such as isobutanol from renewable resources

Impact of Cigarette Smoking Status on Pain Intensity Among Veterans With and Without Hepatitis C

Objective: Chronic pain is a significant problem in patients living with hepatitis C virus (HCV). Tobacco smoking is an independent risk factor for high pain intensity among veterans. This study aims to examine the independent associations with smoking and HCV on pain intensity, as well as the interaction of smoking and HCV on the association with pain intensity. Design/Particpants: Cross-sectional analysis of a cohort study of veterans of Operations Enduring Freedom/Iraqi Freedom/New Dawn (OEF/OIF/OND) who had at least one visit to a Veterans Health Administration (VHA) primary care clinic between 2001 and 2014. Methods: HCV was identified using ICD-9 codes from electronic medical records (EMRs). Pain intensity, reported on a 0-10 numeric rating scale, was categorized as none/mild (0-3) and moderate/severe (4-10). Results: Among 654,841 OEF/OIF/OND veterans (median age [interquartile range] = 26 [23-36] years), 2,942 (0.4%) were diagnosed with HCV. Overall, moderate/severe pain intensity was reported in 36% of veterans, and 37% were current smokers. The adjusted odds of reporting moderate/severe pain intensity were 1.23 times higher (95% confidence interval [CI] = 1.14-1.33) for those with HCV and 1.26 times higher (95% CI = 1.25-1.28) for current smokers. In the interaction model, there was a significant Smoking Status x HCV interaction (P = 0.03). Among veterans with HCV, smoking had a significantly larger association with moderate/severe pain (adjusted odds ratio [OR] = 1.50, P \u3c 0.001) than among veterans without HCV (adjusted OR = 1.26, P \u3c 0.001). Conclusions: We found that current smoking is more strongly linked to pain intensity among veterans with HCV. Further investigations are needed to explore the impact of smoking status on pain and to promote smoking cessation and pain management in veterans with HCV

A whey protein supplement decreases post-prandial glycemia

<p>Abstract</p> <p>Background</p> <p>Incidence of diabetes, obesity and insulin resistance are associated with high glycemic load diets. Identifying food components that decrease post-prandial glycemia may be beneficial for developing low glycemic foods and supplements. This study explores the glycemic impact of adding escalating doses of a glycemic index lowering peptide fraction (GILP) from whey to a glucose drink.</p> <p>Methods</p> <p>Ten healthy subjects (3M, 7F, 44.4 ± 9.3 years, BMI 33.6 ± 4.8 kg/m²) participated in an acute randomised controlled study. Zero, 5, 10 and 20 g of protein from GILP were added to a 50 g glucose drink. The control (0 g of GILP) meal was repeated 2 times. Capillary blood samples were taken fasting (0 min) and at 15, 30, 45, 60, 90 and 120 minutes after the start of the meal and analyzed for blood glucose concentration.</p> <p>Results</p> <p>Increasing doses of GILP decreased the incremental areas under the curve in a dose dependant manner (Pearson's r = 0.48, p = 0.002). The incremental areas (iAUC) under the glucose curve for the 0, 5, 10, and 20 g of protein from GILP were 231 ± 23, 212 ± 23, 196 ± 23, and 138 ± 13 mmol.min/L respectively. The iAUC of the 20 g GILP was significantly different from control, 5 g GILP and 10 g GILP (p < 0.001). Average reduction in the glucose iAUC was 4.6 ± 1.4 mmol.min/L per gram of ingested GILP.</p> <p>Conclusion</p> <p>Addition of GILP to a oral glucose bolus reduces blood glucose iAUC in a dose dependent manner and averages 4.6 ± 1.4 mmol.min/L per gram of GILP. These data are consistent with previous research on the effect of protein on the glycemic response of a meal.</p

RNAcentral 2021: secondary structure integration, improved sequence search and new member databases

RNAcentral is a comprehensive database of non-coding RNA (ncRNA) sequences that provides a single access point to 44 RNA resources and >18 million ncRNA sequences from a wide range of organisms and RNA types. RNAcentral now also includes secondary (2D) structure information for >13 million sequences, making RNAcentral the world's largest RNA 2D structure database. The 2D diagrams are displayed using R2DT, a new 2D structure visualization method that uses consistent, reproducible and recognizable layouts for related RNAs. The sequence similarity search has been updated with a faster interface featuring facets for filtering search results by RNA type, organism, source database or any keyword. This sequence search tool is available as a reusable web component, and has been integrated into several RNAcentral member databases, including Rfam, miRBase and snoDB. To allow for a more fine-grained assignment of RNA types and subtypes, all RNAcentral sequences have been annotated with Sequence Ontology terms. The RNAcentral database continues to grow and provide a central data resource for the RNA community

RNAcentral 2021: secondary structure integration, improved sequence search and new member databases.

RNAcentral is a comprehensive database of non-coding RNA (ncRNA) sequences that provides a single access point to 44 RNA resources and >18 million ncRNA sequences from a wide range of organisms and RNA types. RNAcentral now also includes secondary (2D) structure information for >13 million sequences, making RNAcentral the world's largest RNA 2D structure database. The 2D diagrams are displayed using R2DT, a new 2D structure visualization method that uses consistent, reproducible and recognizable layouts for related RNAs. The sequence similarity search has been updated with a faster interface featuring facets for filtering search results by RNA type, organism, source database or any keyword. This sequence search tool is available as a reusable web component, and has been integrated into several RNAcentral member databases, including Rfam, miRBase and snoDB. To allow for a more fine-grained assignment of RNA types and subtypes, all RNAcentral sequences have been annotated with Sequence Ontology terms. The RNAcentral database continues to grow and provide a central data resource for the RNA community. RNAcentral is freely available at https://rnacentral.org

The 2.5 m Telescope of the Sloan Digital Sky Survey

We describe the design, construction, and performance of the Sloan Digital Sky Survey Telescope located at Apache Point Observatory. The telescope is a modified two-corrector Ritchey-Chretien design which has a 2.5-m, f/2.25 primary, a 1.08-m secondary, a Gascoigne astigmatism corrector, and one of a pair of interchangeable highly aspheric correctors near the focal focal plane, one for imaging and the other for spectroscopy. The final focal ratio is f/5. The telescope is instrumented by a wide-area, multiband CCD camera and a pair of fiber-fed double spectrographs. Novel features of the telescope include: (1) A 3 degree diameter (0.65 m) focal plane that has excellent image quality and small geometrical distortions over a wide wavelength range (3000 to 10,600 Angstroms) in the imaging mode, and good image quality combined with very small lateral and longitudinal color errors in the spectroscopic mode. The unusual requirement of very low distortion is set by the demands of time-delay-and-integrate (TDI) imaging; (2) Very high precision motion to support open loop TDI observations; and (3) A unique wind baffle/enclosure construction to maximize image quality and minimize construction costs. The telescope had first light in May 1998 and began regular survey operations in 2000.Comment: 87 pages, 27 figures. AJ (in press, April 2006