Abstract Insulin is an important biotherapeutic protein, and it is also a model protein used to study amyloid diseases, such as Alzheimer's and Parkinson's. The preparation of the protein can lead to small amounts of aggregate in the solution, which in turn may lead to irreproducible in vitro results. Using several pre‐treatment methods, we have determined that pH cycling and diafiltration of the insulin removes microaggregates that may be present in the solution. These microaggregates were not detectable with traditional biochemical methods, but using small‐angle neutron scattering, we were able to show that pH cycling reduces the radius of gyration of the insulin. Diafiltration removes the aggregates by size and pH cycling dissolves the aggregates by adjusting the pH through the pI of the protein. Pre‐treating the insulin with either pH cycling or diafiltration allowed reproducible kinetics of fibrillation for the insulin protein. Microaggregates are a common problem in protein production, formulation, and preparation; here we show that they are the main cause for inconsistent behavior and how pH cycling and diafiltration can mitigate this problem. Biotechnol. Bioeng. 2011; 108:237–241. © 2010 Wiley Periodicals, Inc.
Abstract BACKGROUND: The efficacy of common fungicides to control downy mildew ( Plasmopara viticola ) in vines decreases when the same commercial formulations are applied over several years. For this reason, new commercial formulations (containing new fungicides) have to offer to grape‐growers to increase the efficacy of antifungal treatments. The efficacy of five new commercial formulations (Cabrio Top, Equation Pro, Fantic M, Mikal Premium and Fobeci) to control downy mildew in an experimental vineyard producing white grapes ( Vitis vinifera cv. Godello) was studied. All these products have in common a new fungicide (pyraclostrobin, famoxadone, benalaxyl‐M, iprovalicarb and benalaxyl, respectively) in their chemical formulation. RESULTS: Two commercial formulations, Cabrio Top and Fobeci, gave the highest efficacy to control this fungal disease. Once treated white grapes were harvested, the fate of these fungicides from vine to wine was studied. The white winemaking process allowed the dissipation of fungicide residues in high percentages (∼95%), except for iprovalicarb (45%). Concentrations in final filtered white wines were close to or lower than 0.01 mg L −1 for all detected fungicides except iprovalicarb and fludioxonil. The following dissipation rate was found: famoxadone > cyprodinil > pyraclostrobin > benalaxyl > folpet > fludioxonil > benalaxyl‐M > penconazole > iprovalicarb. CONCLUSION: From the data obtained in the vinification process, estimated maximum residue levels for white wines were proposed for future EU legislation to control the presence of pesticides in white wines. Thereby one helps to assure also the wine quality and protection of consumer health. Copyright © 2009 Society of Chemical Industry
An ion chromatographic (IC) method for the determination of six organic acids and three inorganic anions in Bayer liquors was proposed. Formic, acetic, propionic, oxalic, succinic, glutaric acid, F−, Cl−, and SO 4 2− were separated and determined within 33 min. For the first time, repeatability, reproducibility, and recoveries for the determination of these acids in Bayer liquors were estimated. The analytes were removed from a Bayer liquor by using an ion-exchange resin column. The chromatographic separation was achieved with only one IonPac AS11-HC column thermostated at 30°C. Organic acids and inorganic anions were detected with a suppressed conductance detector. The precision results showed that the relative standard deviations of the repeatability and reproducibility were <2.94 and <1.37%, respectively. The accuracy of the method was confirmed with an average recovery ranging between 86.3 and 105.6%. Under optimum conditions the detection limits ranged from 0.008 to 0.053 mg/L.
The genetic management of small populations, irrespective of the objective pursued, it is conditioned by their census. Actually the distinction between conservation and selection programmes tends to vanish because in both types of programmes attention should be paid to the increase of inbreeding and to the maintenance or improvement of some traits of interest. To maintain genetic diversity the criterion to follow is to maximize the effective population size. For this purpose several simple rules have been developed during the last decades although there exist also a sophisticated solution, probably optimal. For selection programmes there are also simple rules and an optimal solution although now the desired genetic progress should be included. The design of optimal mating systems, although it presents more problems from a theoretical point of view, it has also a reasonable practical solution.
Preeclampsia and gestational hypertension are major contributors to perinatal morbidity and mortality. Several studies aimed to test the effects of low-dose aspirin (ASA) in the prevention of preeclampsia concluded that the beneficial effects of such treatment outweigh adverse ones. Such benefits have not been fully corroborated by larger randomized trials usually carried out in low-risk women, testing a dose of 60 mg/d ASA presumably ingested in the morning, and including women randomized as late as at 26-32 wks of gestation. The authors conducted a prospective, randomized, double-blind, placebo-controlled, chronotherapy trial on 350 high-risk pregnant women (183 nulliparous), 30.7 ± 5.3 (mean ± SD) yrs of age, and 13.5 ± 1.4 wks of gestation at the time of recruitment. Women were randomly assigned to one of six groups, defined according to treatment (placebo or ASA, 100 mg/d) and time of treatment: upon awakening, 8 h after awakening, or at bedtime. Intervention started at 12-16 wks of gestation and continued until delivery. Blood pressure (BP) was measured by ambulatory monitoring (ABPM) for 48-h at baseline, every 4 wks until the 7th month of gestation, every 2 wks thereafter until delivery, and at puerperium. The effects of ASA on ambulatory BP were markedly dependent on administration time: there was no effect on BP, compared with placebo, when ASA was ingested upon awakening, but the BP reduction was highly statistically significant when low-dose ASA was ingested 8 h after awakening and, to a greater extent, at bedtime (p < .001). At puerperium, 6-8 wks after discontinuation of treatment, there was no statistically significant difference in 24-h BP means between the groups of women who ingested ASA at different circadian times. Women ingesting low-dose ASA, compared with placebo, evidenced a significantly lower hazard ratio (HR) of serious adverse outcomes, a composite of preeclampsia, preterm delivery, intrauterine growth retardation (IUGR), and stillbirth (.35, 95% confidence interval [CI]: .22-.56; p < .001). The HR of individual outcome variables, i.e., preeclampsia, preterm delivery, IUGR, and gestational hypertension, were also significantly lower with ASA versus placebo (p always < .041). There were small and nonsignificant differences in outcomes between placebo and low-dose ASA ingested upon awakening. These four groups combined showed highly significant greater event rate of serious adverse outcomes than women ingesting ASA either in the evening or at bedtime (HR: .19, 95% CI: .10-.39; p < .001). There was no increased risk of hemorrhage, either before or after delivery, with low-dose ASA relative to placebo (HR: .57, 95% CI: .25-1.33; p = .194). Results indicate that (i) 100 mg/d ASA should be the recommended minimum dose for prevention of complications in pregnancy; (ii) ingestion of low-dose ASA should start at ≤16 wks of gestation; and (iii) low-dose ASA ingested at bedtime, but not upon awakening, significantly regulates ambulatory BP and reduces the incidence of preeclampsia, gestational hypertension, preterm delivery, and IUGR. ABPM evaluation at the first trimester of pregnancy provides sensitive endpoints for identification of women at high risk for preeclampsia who might benefit most from the cost-effective preventive intervention with timed low-dose ASA.
Abstract — — Studies based on casual blood pressure measurements concluded that both age and parity have significant effects on blood pressure during pregnancy. We have tested these results on clinically healthy normotensive women who were systematically studied by ambulatory blood pressure monitoring during their pregnancies. We analyzed 1254 blood pressure series sampled for 48 consecutive hours every 4 weeks from the first obstetric visit (usually within the first trimester of pregnancy) until delivery in 205 normotensive pregnant women. Data were divided for comparative analysis by parity (nulliparous versus multiparous), age (≤25, 26 to 30, 31 to 35, and ≥36 years), and trimester of gestation. Circadian parameters established by population multiple-component analysis were compared between groups with a nonparametric test. Effects of age and parity on blood pressure were also tested by ANOVA. A highly statistically significant circadian pattern described by a model that includes components with periods of 24 and 12 hours is demonstrated for systolic and diastolic blood pressure for all groups of pregnant women in all trimesters (always P <0.001). There was no significant difference in 24-hour mean among groups divided by parity at any age or stage of pregnancy (always P >0.160). A trend of increasing blood pressure with age was found for diastolic but not systolic blood pressure. Although statistically significant, differences in the 24-hour mean of diastolic blood pressure among groups divided by age were always <1.5 mm Hg. Data obtained from systematic ambulatory monitoring in normotensive pregnant women indicate the lack of differences in blood pressure according to parity. The small, although significant, increase in diastolic blood pressure with age may have little influence in the proper identification of women with gestational hypertension. Reference thresholds for blood pressure to be used in the early identification of hypertensive complications in pregnancy could thus be developed as a function of rest-activity cycle and gestational age, independent of parity or maternal age.
Administration of valsartan at bedtime as opposed to upon wakening improves the sleep-time relative blood pressure (BP) decline towards a more normal dipper pattern without loss of 24-h efficacy. Amlodipine, however, has been shown to be effective in reducing BP throughout the day and night, independent of dosing time. A large proportion of hypertensive subjects cannot be properly controlled with a single medication. However, no study has yet investigated the potential differing effects of combination therapy depending of the time-of-day of administration. Accordingly, the authors investigated the administration-time-dependent BP-lowering efficacy of valsartan/amlodipine combination. The authors studied 203 hypertensive subjects (92 men/111 women), 56.7 +/- 12.5 yrs of age, randomized to receive valsartan (160 mg/day) and amlodipine (5 mg/day) in one of the following four therapeutic schemes: both medications on awakening, both at bedtime, either one administered on awakening and the other at bedtime. BP was measured by ambulatory monitoring for 48 consecutive hours before and after 12 wks of treatment. Physical activity was simultaneously monitored every min by wrist actigraphy to accurately determine the beginning and end of daytime activity and nocturnal sleep. BP-lowering efficacy (quantified in terms of reduction of the 48-h mean of systolic/diastolic BP) was highest when both hypertension medications were ingested at bedtime, as compared to any one of the three other tested therapeutic schemes (17.4/13.4 mm Hg reduction with both medications on awakening; 15.1/9.6 mm Hg with valsartan on awakening and amlodipine at bedtime; 18.2/12.3 mm Hg with valsartan at bedtime and amlodipine on awakening; 24.7/13.5 mm Hg with both medications at bedtime; p < .018 between groups). The sleep-time relative BP decline was significantly increased towards a more normal dipper pattern only when both medications were jointly ingested at bedtime (p < .001). Bedtime dosing of the combination of the two medications also resulted in the largest percentage of controlled subjects among all the assessed therapeutic schemes (p = .003 between groups). In subjects requiring combination therapy to achieve proper BP control, the association of amlodipine and valsartan efficiently reduces BP for the entire 24 h independent of dosing time. However, the greater proportion of controlled patients, improved efficacy on lowering asleep BP mean, and increased sleep-time relative BP decline suggest valsartan/amlodipine combination therapy should be preferably administered at bedtime.
Abstract Myristica fragrans (Houtt.) is an evergreen tree native to the Maluku Islands, Indonesia. M . fragrans kernel is extensively used in Indian traditional medicines to treat various diseases. Several studies attempt to compile and interpret the pharmacological potential of Myristica fragrans (Houtt.) aqueous and various chemical extracts. Thus, the pharmacological potential of nutmeg essential oil has not been reviewed phytochemically and pharmacologically. Therefore, the present study aimed to share appropriate literature evidence regarding the plant essential oil chemical composition and therapeutic potential of Myristica fragrans essential oil (MFEO). MFEO of leaf, mace, kernel, and seed were used worldwide as potential Ayurvedic medicine and fragrance. MFEO extracted by various methods and oil yield was 0.7–3.2, 8.1–10.3, 0.3–12.5, and 6.2–7.6% in leaf, mace, seed, and kernel. The primary chemical constituents of MFEO were sabinene, eugenol, myristicin, caryophyllene, β‐myrcene, and α‐pinene. Clinical and experimental investigations have confirmed the antioxidant, antimicrobial, antiinflammatory, anticancer, antimalarial, anticonvulsant, hepatoprotective, antiparasitic, insecticidal, and nematocidal activities of MFEO. It is the first attempt to compile oil yield, composition, and the biological activities of MFEO. In future, several scientific investigations are required to understand the mechanism of action of MFEO and their bioactive constituents.
The main aim of the present work was to evaluate the suitability of recycled paperboard coated on its internal face for use in containers that contact fried foods. Suitability was assessed in terms of overall migration into fatty food simulants and in terms of the residue extracted into n-heptane. Testing of the packaging was carried out following the guidelines laid down by the EU for plastic packaging, and also those established by the FDA for the extractive testing of paper and paperboard for use in food contact. With a view to simplifying the test procedures, the results of the official tests were compared with those obtained using alternative fatty food simulants under comparable sets of test conditions. The overall migration test using olive oil as the fatty food simulant was an adaptation of the European Committee for Standardization (CEN) test developed for plastic materials. Two methods for determination of the amount of olive oil absorbed by the paperboard during the tests were compared: a gravimetric method and a modification of the CEN gas chromatographic method. Finally, the packaging materials were extracted with solvents of several different polarities in order to develop a test allowing preliminary identification of major potential migrating components in paperboard contributing to EU overall migration levels and FDA extractive levels. Keywords: Food safety; recycled paperboard coated for food contact; EU overall migration tests; FDA extractive tests; fatty food contact
Abstract Multicolor fluorescent containers : A silica coating procedure has been devised for the preparation of multifluorescent dye‐loaded zeolite L crystals with luminescent anisotropic cores and isotropic shells. The method comprises a combination of the polyelectrolyte layer‐by‐layer technique and Stöber synthesis (see scheme, TEM image, and fluorescence microscopy images). magnified image
