Genetic engineering of Bacillus subtilis for the commercial production of riboflavin

Recombinant DNA engineering was combined with mutant selection and fermentation improvement to develop a strain of Bacillus subtilis that produces commercially attractive levels of riboflavin. The B. subtilis riboflavin production strain contains multiple copies of a modified B. subtilis riboflavin biosynthetic operon (rib operon) integrated at two different sites in the B. subtilis chromosome. The modified rib operons are expressed constitutively from strong phage promoters located at the 5′ end and in an internal region of the operon. The engineered strain also contains purine analog-resistant mutations designed to deregulate the purine pathway (GTP is the precursor for riboflavin), and a riboflavin analog-resistant mutation in ribC that deregulates the riboflavin biosynthetic pathway. Received 22 June 1998/ Accepted in revised form 6 November 1998     

Molecular dynamics of synthetic leucine-serine ion channels in a phospholipid membrane

Molecular dynamics calculations were carried out on models of two synthetic leucine-serine ion channels: a tetrameric bundle with sequence (LSLLLSL)(3)NH(2) and a hexameric bundle with sequence (LSSLLSL)(3)NH(2). Each protein bundle is inserted in a palmitoyloleoylphosphatidylcholine bilayer membrane and solvated by simple point charge water molecules inside the pore and at both mouths. Both systems appear to be stable in the absence of an electric field during the 4 ns of molecular dynamics simulation. The water motion in the narrow pore of the four-helix bundle is highly restricted and may provide suitable conditions for proton transfer via a water wire mechanism. In the wider hexameric pore, the water diffuses much more slowly than in bulk but is still mobile. This, along with the dimensions of the pore, supports the observation that this peptide is selective for monovalent cations. Reasonable agreement of predicted conductances with experimentally determined values lends support to the validity of the simulations.     

Total fertilization failure and idiopathic subfertility

Background  

To gain more insight in whether failure of intrauterine insemination (IUI) treatment in patients with idiopathic subfertility could be related to diminished fertilization, the aim of this study is to compare the fertilization of an initial IVF procedure after six cycles of IUI and the fertilization of an initial IVF procedure without preceding IUI cycles in couples with idiopathic subfertility.     

Uncertainty in ensemble forecasting of species distribution

Species distribution modelling has been widely applied in order to assess the potential impacts of climate change on biodiversity. Many methodological decisions, taken during the modelling process and forecasts, may, however, lead to a large variability in the assessment of future impacts. Using measures of species range change and turnover, the potential impacts of climate change on French stream fish species and assemblages were evaluated. Our main focus was to quantify the uncertainty in the projections of these impacts arising from four sources of uncertainty: initial datasets (Data), statistical methods [species distribution models (SDM)], general circulation models (GCM), and gas emission scenarios (GES). Several modalities of the aforementioned uncertainty sources were combined in an ensemble forecasting framework resulting in 8400 different projections. The variance explained by each source was then extracted from this whole ensemble of projections. Overall, SDM contributed to the largest variation in projections, followed by GCM, whose contribution increased over time equalling almost the proportion of variance explained by SDM in 2080. Data and GES had little influence on the variability in projections. Future projections of range change were more consistent for species with a large geographical extent (i.e., distribution along latitudinal or stream gradients) or with restricted environmental requirements (i.e., small thermal or elevation ranges). Variability in projections of turnover was spatially structured at the scale of France, indicating that certain particular geographical areas should be considered with care when projecting the potential impacts of climate change. The results of this study, therefore, emphasized that particular attention should be paid to the use of predictions ensembles resulting from the application of several statistical methods and climate models. Moreover, forecasted impacts of climate change should always be provided with an assessment of their uncertainty, so that management and conservation decisions can be taken in the full knowledge of their reliability.     

A forty-five year investigation for secular changes in physical maturation

In a stable, economically- and educationally-privileged population of 180 subjects observed from birth to beyond the period of maximum growth in height, no secular changes toward larger size or earlier adolescence were observed between the earlier-born and later-born boys or girls of the Child Research Council study series. The mid-birthdate for the 45 years of data collection was January 1, 1940 for the girls and May 1, 1936 for the boys. With none of the mean differences significant at the 0.05 level of confidence, the earlier-born subjects were both slightly taller and heavier from birth and, for the girls, adolescence was slightly earlier. Forty pairs of like-sexed siblings were included in the data. Coefficients of correlation were statistically significant for birthweight and menarcheal ages for the sisters and for height and weight at the age of maximum increment of growth in height for both sexes. In 12 of the 16 pairs of sisters, the older menstruated at an earlier age than the younger.     

家庭氧疗配合呼吸操治疗慢性阻塞性肺疾病患者的疗效

目的：观察长期家庭氧疗配合呼吸操治疗慢性阻塞性肺疾病（COPD）的临床效果。、方法：选择本院收治的COPD患者80例,随机分为研究组和对照组,各40例,两组均给予常规治疗,研究组同时由医务人员指导给予家庭氧疗和呼吸操训练,随访1年．观察两组治疗前、治疗后6个月和1年肺功能,应用StGeorge’S呼吸问卷（SGRQ）评价生存质量、结果：研究组1年内急性发作1次5人,2次2人,3次及以上1人,对照组1年内急性发作1次9人,2次7人,3次及以上4人（P〈0．05）;研究组治疗后6个月、1年用力肺活量（FVC）、第1秒用力呼气容积（FEVl）和FEVl／FVC均明显升高（P〈0．05）,对照组治疗后6个月、1年FVC、FEVI／FVC与治疗前比较无统计学差异（P〉0．05）;研究组治疗后6个月、1年SGRQ明显降低（P〈0．05）,对照组治疗后6个月、1年SGRQ与治疗前比较无统计学差异（P〉0．05）．结论：家庭氧疗配合呼吸操能明显改善COPD患者肺功能,延缓患者肺功能恶化,提高患者生存质量。     

血清胆红素水平与缺血性卒中的相关性研究

目的：脑卒中是威胁人类健康三大疾病之一,是我国成人致残的首要原因,其中80％是缺血性卒中。本文意在研究血清胆红素水平与缺血性卒中严重程度、发病机理以及颈动脉粥样硬化斑块的关系,以进一步为防治缺血性卒中的发生、发展提供新的途径。方法：选择缺血性脑卒中患者（观察组）150例和同期健康体检者（对照组）150例,分别测定两组的血清总胆红素水平（TBIL）、间接胆红素（DBIL）、直接胆红素（IBIL）,并对病例组进行TOAST分型、NIHSS评分及颈部血管超声检查。比较两组间血清胆红素,及观察组内不同分型组间血清胆红素的差异。结果：缺血性卒中患者TBIL、DBIL水平显著高于正常对照组,差异具有统计学意义（P〈0．05）。缺血性卒中患者按TOAST分型各亚型间血清TBIL、DBIL、IBIL水平差异无统计学意义（P〉0．05）。中重型脑梗死组与轻型脑梗死组比较,血清TBIL、DBIL、IBIL浓度均明显升高,差别具有统计学意义（P〈0．05）。缺血性卒中患者中有动脉粥样硬化斑块形成组血清TBIL、DBIL水平低于颈部动脉内膜光滑、完整者组,差别有统计学意义（P〉0．05）。结论：缺血性卒中患者血清胆红素升高,参与急性应激反应,可能作为衡量缺血性卒中严重程度的指标之一;高水平血清胆红素可能预防颈动脉粥样硬化斑块的形成,从而预防缺血性卒中的发生。     

Appearance of D2O in sweat after oral and oral-intravenous rehydration in men

Intravenous (IV) rehydration is common in athletics, but its thermoregulatory benefits and ergogenicity have not been elucidated. Availability of orally ingested fluid is dependent on gastric emptying and intestinal absorption rate. Deuterium oxide (D2O) has been used to demonstrate that fluid ingested during exercise appears in sweat within 10 minutes. The purpose of this study was to determine the effect of concurrent IV rehydration on D2O appearance in sweat samples after per ora rehydration with D2O labeled fluid. We hypothesized that the combination method would not be superior to the oral method. Ten fit men (age 23 ± 4, VO2max 59.49 ± 4.09 L·min(-1)) underwent 20 hours of fluid restriction resulting in 1.95 ± 0.25% body weight loss before beginning treadmill exercise and cycling. Exercise was performed in an environmental chamber (35.6 ± 0.2° C, 35.0 ± 1.8% relative humidity) for 2 hours at 55% VO2max, and the participants exhibited a mean body weight deficit of 4.50 ± 0.04%. Thermoregulatory measures were recorded while subjects were rehydrated with oral (OR) or oral combined with intravenous (IVO) fluid traced with D2O. After 30 minutes of rehydration and 30 minutes of seated recovery, the subjects began treadmill exercise at 55-60% VO2max. Forehead sweat samples were collected 0, 5, 10, 20, and 75 minutes from the start of rehydration. The samples were analyzed for D2O via isotope ratio mass spectrometry. D2O did not appear in the sweat within 20 minutes of rehydration; however, it did appear during the subsequent exercise bout. There was no significant difference between rehydration modes. Plasma volume increases and decreased volume of orally ingested fluid did not significantly alter transit time from ingestion to appearance in excreted sweat. The IVO method does not appear to be superior to the traditional OR method of rehydration.     

Comparison of body cooling methods on physiological and perceptual measures of mildly hyperthermic athletes

Hyperthermia is common among athletes and in a variety of environments. The purpose of this study was to evaluate the effectiveness of cooling methods on core body temperature, heart rate (HR), and perceptual readings in individuals after exercise. Sixteen subjects (age: 24 ± 6 years, height: 182 ± 7 cm, weight: 74.03 ± 9.17 kg, and body fat: 17.08 ± 6.23%) completed 10 exercise sessions in warm conditions (WBGT: 26.64 ± 4.71°C) followed by body cooling by 10 different methods. Cooling methods included cold water immersion (CWI), shade, Port-a-Cool? (FAN), Emergency Cold Containment System? (ECCS), Rehab. Hood? (HOOD), Game Ready Active Cooling Vest? (GRV), Nike Ice Vest? (NIV), ice buckets (IBs), and ice towels (IT). These cooling modes were compared with a control (SUN). Rectal temperature (T(re)), HR, thermal sensation, thirst sensation, and a 56-question Environmental Symptoms Questionnaire (ESQ) were used to assess physiological and perceptual data. Average T(re) after exercise across all trials was 38.73 ± 0.12°C. After 10 minutes of cooling, CWI (-0.65 ± 0.29°C), ECCS (-0.68 ± 0.24°C), and IB (-0.74 ± 0.34°C) had significantly (p < 0.006) greater decreases in T(re) compared with that in SUN (-0.42 ± 0.15°C). The HR after 10 minutes of cooling was significantly (p < 0.006) lower for CWI (82 ± 15 b·min(-1)), ECCS (87 ± 14 b·min(-1)), and IT (84 ± 15 b·min(-1)) when compared with SUN (101 ± 15 b·min(-1)). The thermal sensation between modalities was all significantly (p < 0.006) lower (CWI: 1.5 ± 0.5; Fan: 3.0 ± 1.0; ECCS: 4.5 ± 1.0; Hood: 4.5 ± 0.5; GRV: 4.0 ± 0.5; NIV: 4.5 ± 1.0; IB: 4.0 ± 1.0; IT: 3.0 ± 1.0) when compared with SUN (5.5 ± 0.5), except for Shade (5.0 ± 1.0). There were no significant differences (p > 0.006) in thirst sensation between modalities. The ESQ scores were significantly (p < 0.006) lower for CWI (1 ± 6), Fan (4 ± 5), and IT (3 ± 8) compared with that for SUN (13 ± 12). In conclusion, when athletes experience mild hyperthermia, CWI, ECCS, and IB resulted in a significantly greater decrease in T(re). These cooling strategies are recommended to decrease T(re) during a brief recovery period between exercise bouts.