Molecular structure of extracellular poly(3-hydroxybutyrate) depolymerase fromAlcaligenes faecalis T1

The amino acid sequence of a peptide containing an active serine was examined with poly(3-hydroxybutyrate) (PHB) depolymerase ofAlcaligenes faecalis T1. The sequence Cys-Asn-Ala-Trp-Ala-Gly-Ser-Asn-Ala-Gly-Lys was obtained. This amino acid sequence around the active serine does not fit any reported sequence of other esterases and proteases. On the other hand, a segment of the amino acid sequence of PHB depolymerase ofA. faecalis was homologous to the type III sequence of fibronectin. Similar sequences have been reported in some type of bacterial chitinase and cellulases, and PHB depolymerase seems to have an overall similarity to these bacterial extracellular hydrolases.     

Molecular analysis of genomic DNA allows rapid,and accurate,prenatal diagnosis of peroxisomal D-bifunctional protein deficiency

Prenatal diagnosis was requested for a couple with a previous child affected by the peroxisomal disorder D-bifunctional protein deficiency. Prior analysis of the D-bifunctional protein cDNA sequence from the propositus had shown that it was missing 22 bp. This was subsequently attributed to a point mutation in the intron 5 donor site (IVS5+1G>C) of the D-bifunctional protein gene. Consistent with parental consanguinity, the patient was shown to be homozygous for this mutation, which is associated with loss of a Hph 1 restriction site in the genomic sequence. Prenatal testing of the fetus using genomic DNA isolated from uncultured amniocytes indicated that both alleles of the D-bifunctional protein had the IVS5+1G>C substitution. The peroxisomal defect was later confirmed biochemically using cultured amniocytes, which were found to have elevated levels of very long chain fatty acids (VLCFA). This is the first report of prenatal diagnosis of D-bifunctional protein deficiency using molecular analysis of genomic DNA. Copyright © 2002 John Wiley & Sons, Ltd.     

Maternal serum human chorionic gonadotrophin and pregnancy-associated plasma protein a,markers for fetal down syndrome at 8–14 weeks

Maternal serum levels of human chorionic gonadotrophin and its subunits (intact, α, and free βhCG) and pregnancy-associated plasma protein A (PAPP-A) were measured in 279 women between 8 and 14 weeks' gestation. This group included 23 pregnancies in which the fetus had Down syndrome (DS), diagnosed either at birth or during the second trimester (n=17) or from chorionic villus sampling (CVS) (n=6). Normal medians were determined from the 258 apparently normal pregnancies. The median levels of intact hCG (1·4 MOM) and free βhCG (2·1 MOM) were significantly raised, whereas the median level of PAPP-A (0·39 MOM) was significantly lower in the DS pregnancies when compared with the control group. Levels of αhCG were similar in both the control and the DS pregnancies. Analysis of samples taken prior to 14 weeks' gestation demonstrated that only PAPP-A (0·34 MOM) was significantly altered in DS pregnancies. However, after the exclusion of DS cases diagnosed at CVS, the median intact hCG (1·56 MOM), free βhCG (2·27 MOM), and αhCG (1·8 MOM) were all raised in DS pregnancies. This emphasizes the problem of the interpretation of biochemical markers when DS cases are diagnosed at CVS.     

Effect of sunlight radiation,rainfall and droplet spectra of sprays on persistence of bacillus thuringiensis deposits after application of dipel® 76af formulation onto conifers

Abstract

The effect of sunlight radiation, rainfall and droplet spectra of sprays on per ‐sistence of a Bacillus thuringiensis subspp. kurstaki (Btk) formulation, DiPel^® 76AF, was examined after application onto spruce [Picea glauca (Moench) Voss] foliage. The investigation consisted of three studies: (i) Study I: a laboratory microcosm study to examine the photostability of DiPel 76AF deposits on foliage after different periods of exposure to two radiation intensities, (ii) Study II: a laboratory microcosm study to examine the rainfastness of foliar deposits after exposure to different amounts of rainfall consisting of two separate droplet spectra, and (iii) Study III: a field microcosm study to investigate the influence of two different droplet spectra of DiPel 76AF sprays on foliar persistence of Btk under natural weathering conditions. In all studies, persistence of Btk was investigated both by bioassay [using spruce budworm (Choristoneura fumiferana Clemens)] and total protein assay.

The findings of Study I indicated that bioactivity of foliar deposits decreased with increasing duration of exposure to radiation, and with increasing radiation intensity. The half‐life (DT₅₀, the exposure period required for 50% of the initial bioactivity to disappear) was 5.1 d for the low intensity, and 3.9 d for the higher intensity. In contrast with the bioassay results, the total protein levels [determined by the bicinchoninic acid (BCA) method] showed no decrease with increasing duration of exposure, or with increasing radiation intensity.

The findings of Study II indicated that bioactivity of foliar deposits decreased with increasing cumulative rainfall. A new term, RF₅₀ [the amount of rain (in mm) required to washoff 50% of the initial deposit], was introduced to understand the relationship between rainfall intensity and reduction in bioactivity. When the same amount of rain was applied in different droplet sizes, the RF₅₀ value was high (5.2 mm) for the small rain droplets, and was low (2.9 mm) for the large rain droplets. Similar to the bioassay results, the total protein concentrations (determined by the BCA method) decreased with increasing amount of rain and with increasing rainfall intensity. The RF₅₀ value (obtained using ng protein /cm²) was 5.4 mm for the small rain droplets, and was 3.4 mm for the large rain droplets.

The field microcosm study indicated that when DiPel 76AF was applied in small droplets (D_v.5 of 65 μm), the persistence of bioactivity was ca 8.0 d, whereas when it was applied in large droplets (D_v.5 of 130 nm) it was ca 11 d. Bioactivity decreased with time after spray, and the DT₅₀ was 1.98 d for the spray of small droplets, and 2.87 d for that of large droplets. Similar to the bioactivity, the total protein concentrations also decreased with time after spray, and the DT₅₀ values for the small and large droplet spectra were 3.45 and 6.07 d respectively.     

Tomato microtubule-associated protein end-binding protein 1 (EB1) mediates tolerance to salt stress北大核心CSCD

The dynamics of microtubules is regulated mainly by microtubule-associated proteins (MAPs) and plays an important role in plant development and response to environmental signals. End-binding protein 1 (EB1) is a MAP specially binding to the microtubule plus end. Blast search of tomato genome showed two EB1 genes, which were named as SlEB1a (Solyc03g116370) and SlEB1b (Solyc02g092950) in this study. Transgenic tomato plants over-expressing SlEB1a or RNA interfering both SlEB1a and SlEB1b were constructed, and their sensitivity to microtubule depolymerization drug propyzamide and salt stress were analyzed. In this study, we determined the role of tomato EB1 (SlEB1) in the response to salt stress. Compared to the wild-type control plants, OE plants were more sensitive to 1 μmol/L propyzamide, whereas RNAi plants were more tolerant to 1 μmol/L propyzamide; in contrast, OE plants were more tolerant to 100 mmol/L NaCl, whereas RNAi plants were more sensitive to 100 mmol/L NaCl. Thus, SlEB1 might positively regulate salt stress response by negatively regulating the dynamics of tomato cortical microtubules. This study forms a basis for how cortical microtubule dynamics plays a role in plant response to salt stress. © 2018 Science Press. All rights reserved.     

Dynamic Mechanical Properties of Soy Protein Filled Elastomers

Dynamic mechanical properties including temperature effect, stress softening, and Payne effect are studied on the elastomer composites filled with soy protein or carbon black. The comparison of protein composite with well-known carbon black composites provides further insight into the protein composites. The elastomers filled with soy protein aggregates give substantial reinforcement effect when compared with the unfilled elastomers. Approximately 400 times increase in shear elastic modulus was observed when 40% by weight of protein is incorporated into the elastomers. The sample films were cast from the particle dispersion of soy protein isolate and carboxylated styrene–butadiene latex. At the higher temperatures, the shear elastic modulus of soy protein-filled composites does not decrease as much as that of the carbon black-filled composites. The behavior of elastic and loss modulus under the oscillatory strain of different magnitude is similar to that of carbon black reinforced styrene–butadiene rubber. However, carbon black composites show a better recovery behavior after eight cycles of dynamic strain. The reduction of shear elastic modulus with dynamic strain (Payne effect) was compared with Kraus model and the fitting parameter related to the aggregate structure of the soy protein. A reasonable agreement between the theoretical model and experiment was obtained, indicating the Payne effect of the protein-related network structure in the elastomers could also be described by the kinetic agglomeration de-agglomeration mechanism.     

Public Health,Ethics, and Functional Foods

Functional foods aim to provide a positive impact on health and well-being beyond their nutritive content. As such, they are likely candidates to enhance the public health official’s tool kit. Or are they? Although a very small number of functional foods (e.g., phytosterol-enriched margarine) show such promise in improving individual health that Dutch health insurance companies reimburse their costs to consumers, one must not draw premature conclusions about functional foods as a group. A large number of questions about individual products’ safety, efficacy, and affordability need to be answered before they might become an important part of the public health agenda. More importantly, though, the costs and benefits of functional foods relative to alternative mechanisms of public health improvement need to be ascertained. Alternative scenarios that warrant investigation are mainly the supply of nutraceutical ingredients in pill form targeting “at risk” groups and consumer education on diet and lifestyle.     

Purification of Aspergillus fumigatus (Pdf1) Poly(β-hydroxybutyrate) (PHB) Depolymerase Using a New, Single-Step Substrate Affinity Chromatography Method: Characterization of the PHB Depolymerase Exhibiting Novel Self-Aggregation Behavior

The extracellular poly(-hydroxybutyrate) (PHB) depolymerase of Aspergillus fumigatus Pdf1 was purified by a new, simple, one-step affinity chromatography method using the substrate PHB. The purified enzyme was glycosylated, with the molecular mass of 40 KD, and exhibited a novel self-aggregation behavior by means of hydrophobic interaction that was resolved by Triton X-100 (TX-100) pretreatment of enzyme and also TX-100 incorporation in the native gel. The apparent K _m value of purified enzyme for PHB was 119 g/mL and 3-hydroxybutyrate was detected as the main endproduct of PHB hydrolysis. The depolymerase was insensitive to phenylmethyl sulfonyl fluoride (PMSF), sodium azide, ethylenediaminetetraacetic acid (EDTA), and para-chloromercuric benzoic acid (PCMB), but was inactivated by dithioerythritol (DTT) and showed specificity for short chain-length poly(-hydroxyalkanoates) (PHAs) such as PHB, poly(hydroxyvalerate) (PHV), and copolymers of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV). Medium-chain-length PHA failed to get hydrolyzed. The enzyme, however, exhibited strong cross reactivity with the Comamonas sp. PHB depolymerase antibodies, but not with PHV depolymerase antibodies of Pseudomonas lemoignei. Southern hybridization and dot blot analysis of A. fumigatus Pdf1 genomic DNA with alkaline phosphatase labeled probes of P. lemoignei PHB and PHV depolymerase genes revealed no homology, although the enzyme hydrolyzed both PHB and PHV.     

Electrosynthesis of κ-Carrageenan Complexes with Gelatin

Electrosynthesis at pH 9, 12, and 11 of -carrageenan–gelatin complexes from electrolytes containing the components at proportions varying from 1:5 to 5:1 provided complexes in the composition varying in a range from 1:10 to 1:1.8. Components were bound mainly electrostatically, although partial insolubility of complexes in 7 M aqueous urea pointed to a part of the complexes bound covalently. None of the performed tests except solubility could confirm formation of covalent complexes. The complexation of -carrageenan to gelatin significantly increased the thermal stability of carrageenan, whereas the thermal stability of gelatin increased rather insignificantly.