Starch-based extruded plastic films and evaluation of their biodegradable properties

Plastic formulations containing up to 40% starch were prepared and blown into thin films using extrusion methods. Extruded films were evaluated for their biodegradability by exposing them to a consortium of starch degrading bacteria in the laboratory for 45 days and in the ‘La Silla’ river located in Monterrey, N.L. Mexico for up to 60 days. Biodegradability was assessed by measuring changes in weight loss and chemical composition of the films using Fourier transform infrared (FTIR) spectroscopy. While little or no degradation was apparent in control films made up of 100% low density polyethylene (LDPE) or 100% poly-(ethylene-co-acrylic acid) (EAA), most of the starch was depleted in starch-containing films exposed in the river. Starch degradation in films exposed to amylolytic bacteria in the laboratory was relatively slower. Also, increasing the amount of EAA from 25% to 50% substantially reduced starch depletion in these films.     

Fate of starch-containing plastic films exposed in aquatic habitats

Plastic films containing cornstarch (40% dry weight) in combination with polyethylene (PE) and polyethylene-co-acrylic acid (EAA) were exposed to river and pond habitats for up to 60 days. Fourier transform infrared (FTIR) spectroscopic analysis of the starch-plastic films indicated that up to about 40% of starch in the films disappeared after 60 days' exposure. PE and EAA components were unchanged during this period. A diverse microbial biofilm quickly accumulated on each of the plastic films in both environments. Although the amount of biofilm accumulation was greater in the river environment than in the pond environment, there was little quantitative difference in the extent of biofilm formation on plastic films composed of PE, EAA, or starch-PE-EAA in either environment. All three types of plastic films were heavily colonized by a midge-fly larva (Dicrotendipes sp.) by 60 days of exposure in the aquatic environments, but only starch-containing plastic films showed any evidence of physical damage that could have been a result of ingestion of the plastic by the larvae (pond>river), suggesting that, in addition to microbial starch degradation, biodisintegration may also be a significant factor influencing the environmental fate of starch-containing plastics.     

A study of cornstarch granule digestion by an unusually high molecular weightα-amylase secreted byLactobacillus amylovorus

The cells ofLactobacillus amylovorus (NRRL B-4540), grown in a medium containing 2% cornstarch as the sole carbon source, secreted an amylase activity that rapidly solubilized cornstarch. Fourier transform infrared (FTIR) spectroscopic analyses showed that 80–90% of starch was consumed by bacteria in a 10-day-old culture medium. The remnant of starch granules digested in the culture medium inoculated with the cells ofL. amylovorus have also lost their characteristic iodine-binding capacity as visualized by starch dye-binding microplate assay and light microscopy. Scanning electron miscroscopy (SEM) of granules from a 48-h culture medium showed hollow and fragmented granules with a pitting phenomenon characteristically produced by-amylase activity. Analysis of an enzyme preparation from a culture medium ofL. amylovorus revealed that the putative enzyme appears to be a single protein band of unusually high M_r (150,000) on SDS gels stained for amylase activity. Analysis of starch digestion products by thin layer chromatography (TLC) showed enzyme activity typical of-amylase.The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

Starch conversion by amylases fromAureobasidium pullulans

Summary A color variant strain (NRRL Y-12974) ofAureobasidium pullulans produced a saccharifying -amylase and two forms of glucoamylase extracellularly when grown on starch at 28°C for 4 days. A sugar syrup containing DP1 (degree of polymerization) and DP2 (31) was made from maltodextrin DE (dextrose equivalent) 10 (35%, w/w) at 55°C and pH 4.5 using the amylase preparation (40 U g^–1 DS (dry substance). The syrup composition was highly dependent upon substrate concentration but nearly independent of enzyme dose. Glucose syrup containing 93% glucose was made from maltodextrin DE 10 (35%, w/w) at 65°C and pH 4.5 using the same enzyme preparation at 100 U g^–1 DS. The enzyme preparation (100 U g^–1 DS) produced 98–100% glucose from raw corn starch at pH 4.5 and 50°C.The mention of firm names or trade products does not imply that they are endorsed or recommended by the US Department of Agriculture over other firms or similar products not mentioned. Abbreviations: DE, dextrose equivalent (an indication of polymerization; reducing sugars as percentage glucose); DP, degree of polymerization; DP1, glucose; DP2, disaccharide; DP3, trisaccharide; DP4, tetrasaccharide; DS, dry substance.     

Prehension synergies during nonvertical grasping,II: Modeling and optimization

This study examines various optimization criteria as potential sources of constraints that eliminate (or at least reduce the degree of) mechanical redundancy in prehension. A model of nonvertical grasping mimicking the experimental conditions of Pataky et al. (current issue) was developed and numerically optimized. Several cost functions compared well with experimental data including energylike functions, entropylike functions, and a motor command function. A tissue deformation function failed to predict finger forces. In the prehension literature, the safety margin (SM) measure has been used to describe grasp quality. We demonstrate here that the SM is an inappropriate measure for nonvertical grasps. We introduce a new measure, the generalized safety margin (GSM), which reduces to the SM for vertical and two-digit grasps. It was found that a close-to-constant GSM accounts for many of the finger force patterns that are observed when grasping an object oriented arbitrarily with respect to the gravity field. It was hypothesized that, when determining finger forces, the CNS assumes that a grasped object is more slippery than it actually is. An operative friction coefficient of approximately 30\% of the actual coefficient accounted for the offset between experimental and optimized data. The data suggest that the CNS utilizes an optimization strategy when coordinating finger forces during grasping.     

Production and characteristics of an intracellularα-glucosidase from a color variant strain ofAureobasidium pullulans

Aureobasidium pullulans produced an intracellular-glucosidase. The enzyme was purified 124-fold by solubilization with Triton X-100, Q-Sepharose treatment, hydroxylapatite, octyl-Sepharose column chromatography, and gel filtration on Sephacryl S-200, and had a specific activity of 316.82 U/mg protein. The enzyme displayed an optimum pH for its action at 4.0 and was fully stable at pH 3.0–6.0 at 50°C. The-glucosidase was completely stable up to 60°C and had an optimum activity at 60°C. The partially purified enzyme preparation hydrolyzed maltose, isomaltose, sucrose, and trehalose at relative rates of 100, 60, 47, and 50, respectively, and had little or no activity on polysaccharides. TheK _m value for maltose hydrolysis at pH 4.0 and 50°C was 1.85mm. The enzyme was not adsorbed onto raw corn starch and showed little raw starch degradation. The-glucosidase did not require any metal ion for activity. This represents the first characterization of intracellular-glucosidase fromA. pullulans.The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

Enzymatic activities and infrared studies of glutamate dehydrogenase immobilized on Langmuir-Blodgett films

Summary A technique is described for the immobilization of active glutamate dehydrogenase (GDH) on behenic acid Langmuir-Blodgett (LB) films. The optimization of the immobilization conditions shows that the activities of GDH bound on hydrophobic and hydrophilic LB films were similar and decreased dramatically when the immobilized enzyme was dried. The GDH binding was followed by Fourier transform infrared (FTIR) spectroscopy. Modifications of GDH conformation and LB film structure were observed during the enzyme binding. After GDH activity test, a partial dissociation of behenic acid occurred and the -sheet band of the enzyme increased by comparison with the -helix band.Abbreviations LB Langmuir-Blodgett - FTIR spectroscopy Fourier transform infrared spectroscopy - GDH glutamate dehydrogenase - TEA triethylamine     

Ultrastructure of the tube foot sensory-secretory complex in Ophiocomina nigra (Echinodermata,Ophiuridea)

Summary The ultrastructure of the epidermal layer of both the oral and arm podia of the brittle star Ophiocomina nigra is described. Despite external differences, little variation occurs in their internal structure. The podial epidermis, which is overlain by a three-layered cuticle, consists of five cell types: support, mucous, sensory, adhesive secretory and monociliated neurosecretory-like cells. Areas of specialisation are superimposed on this basic plan. These comprise four cells forming cohesive units, made up of two adhesive secretory, one sensory and one monociliated neurosecretory-like cells. The two adhesive secretory cells may be identical or vary in the structure of their secretory packets. The sensory cells are of the normal type bearing a short cilium with a 9+2 microtubular arrangement. The monociliated neurosecretory-like cells contain many small dense vesicles and a short sub-cuticular cilium of irregular microtubular structure. Together, they appear to form a sensory-secretory complex which functions in adhesion both for feeding and locomotion. A system in which the secretion of the monociliated neurosecretory-like cell may control adhesive secretion is proposed.     

Glycerol-plasticized films prepared from starch—poly(vinyl alcohol) mixtures: effect of poly(ethylene-co-acrylic acid)

Equations were obtained from response surface models to show how the ultimate tensile strength (UTS) and percent elongation at break (%E) of solution-cast films vary with relative amounts of starch, poly(vinyl alcohol) (PVA), poly(ethylene-co-acrylic acid) (EAA) and glycerol in the formulation. Equations found from the response surface methodology were used to optimize the relative amounts of the four components with respect to the physical properties of cast films. The model showed that only glycerol content was important to predict the UTS of the films. The model for %E was more complicated, since there was a three-way interaction between EAA, PVA and glycerol. This model also contained two other terms: a two-way interaction evolving glycerol and EAA. and a (PVA)³ term. In general, %E increased as EAA, PVA and glycerol were increased together. However, increased amounts of EAA could decrease %E if EAA was the only component increased. It is believed that EAA forms complexes with both starch and PVA, thereby increasing compatibility of the two polyhydroxy polymers. As %E increases, UTS of the films decreases. All the films produced in this paper were made with starch contents above 50% to insure an optimum film formulation with at least 50% starch. A mixture of 55·6% starch, 2·8% EAA, 28·3% PVA and 13·3% glycerol is believed to be close to the optimum formulation to obtain films having at least 100%E and UTS of 25 MPA, while still maintaining starch concentrations above 50%.     

Quality and biochemical effects of a IBL/IRS wheat-rye translocation in wheat

Wheat (Triticum aestivum L.) breeders world-wide have used rye (Secale cereale L.) as a source of genes for agronomic improvement. The 1BL/1RS wheat-rye chromosomal translocation derived from the Russian cultivars Kavkaz and Aurora has been among the most common means of accessing useful rye genes. Unfortunately, deleterious wheat quality effects are often associated with the presence of 1RS. The identification of genetic backgrounds capable of alleviating the deleterious effects of 1RS is crucial for its continued exploitation. End-use quality parameters and flour protein composition, as measured by size-exclusion high-performance liquid chromatography (SE-HPLC) of 373 wheat lines, derived from seven 1BL/1RS breeding populations, were analyzed. In all populations, significant quality defects were detected in 1BL/1RS lines compared to non-1RS sister lines. The detrimental quality effects resulted from alteration of the ratio of flour protein composition, especially, decreased glutenin concentrations, and increased salt-water soluble protein concentrations. The end-use quality of 1BL/1RS lines, however, was highly dependent on genetic backgrounds. The potential exists for improvement in quality through crosses between 1RS lines with high glutenin, or low salt-water soluble protein concentrations, and non-1RS lines with strong dough properties.Joint contribution of the U.S. Department of Agriculture, Agricultural Research Service and Department of Agronomy, University of Nebraska-Lincoln, as Journal Series Paper No. 10598. Mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture or the University of Nebraska over other firms or products not mentioned     

Pullulan production by color variant strains ofAureobasidium pullulans

Summary Naturally occurring color variant strains ofAureobasidium pullulans are distinguished from typical strains by their brilliant pigmentation, overproduction of secreted enzymes (xylanase), and low DNA relatedness. Color variants have not previously been examined for pullulan secretion. Among five independently isolated color variants, strains NRRL Y-12,974 and YB-4026 made the greatest amounts of pullulan from cornstarch, with conversion efficiencies of about 10%. Neither color variant nor typical strains made significant amounts of pullulan from the unconventional lactose or xylan substrates. Pullulan yields were inversely correlated with biomass production. Pullulan production thus appears to be a variable characteristic of both color variant and typically pigmented strains ofA. pullulans, regulated by specific inducers during growth limitation.The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

Starch characteristics in cultures of normal and mutant maize endosperm

Summary Vigorously growing suspension cultures of normal, amylose-extender (ae) and waxy (wx) maize endosperm were established from near isogenic lines of maize inbred A636. The recovery of the ability to produce vigorous cultures of ae and wx endosperm by backcrossing demonstrate the genetic control of endosperm growth in vitro. Phenotypic expression of the endosperm mutants in culture was studied by examining the properties of starch accumulated in endosperm cultures and starch from developing and mature kernels of the same genotype. After 9 months in culture, the amylose contents of the starch in normal callus tissue and normal endosperm tissue were not significantly different, 28.2% and 31.7%, respectively. Starch granules from normal cultures and endosperm stained blue-black with iodine and were round to polygonal in shape. The starches of wx endosperm and callus cultures contained no amylose, and wx starch granules stained brown-orange with iodine. Although, wx starch granules were primarily round, a few granules with jagged edges were observed in starch samples isolated from cultures and kernels. The percent amylose in starch from ae callus was significantly lower than the amylose content of starch from ae endosperm tissue, 39.9% and 67.7%, respectively. Starch granules from ae endosperm and cultures were smaller than normal and wx starch granules. Irregular starch granules which are typical of ae endosperm were present in ae callus tissue, but were less frequently observed. We conclude that specific endosperm mutant phenotypes are expressed in vitro.Supported in part by the United States Department of Agriculture Competitive Grant 85-CRCR-1-1740. Contribution No. 94, Department of Horticulture. The Pennsylvania State University. Authorized for publication as paper No. 7373 in the journal series of the Pennsylvania Agricultural Experiment Station     

Solution Structures of Casein Peptides: NMR, FTIR, CD, and Molecular Modeling Studies of αs1-Casein, 1–23

To determine its potential for interacting with other components of the casein micelle, the N-terminal section of bovine _s1-casein-B, residues 1-23, was investigated with nuclear magnetic resonance (NMR), Fourier transform infrared (FTIR) and circular dichroism (CD) spectroscopies, and molecular modeling. NMR data were not consistent with conventional -helical or -sheet structures, but changes in N-H proton chemical shifts suggested thermostable structures. Both CD and FTIR predicted a range of secondary structures for the peptide (30–40% turns, 25–30% extended) that were highly stable from 5°C to 25°C. Other conformational elements, such as loops and polyproline II helix, were indicated by FTIR only. Molecular dynamics simulation of the peptide predicted 32% turns and 27% extended, in agreement with FTIR and CD predictions and consistent with NMR data. This information is interpreted in accord with recent spectroscopic evidence regarding the nature of unordered conformations, leading to a possible role of _s1-casein (1–23) in facilitating casein-casein interactions.     

Callus formation from protoplasts of a maize cell culture

Summary A finely dispersed cell suspension culture from the friable callus of the Black Mexican Sweet line of maize was obtained. Protoplasts from this cell culture, when grown in a simplified medium described here, showed sustained cell divisions and gave rise to callus.Abbreviations 2,4-D 2,4-dichlorophenoxyaceticacid - SDS sodium dodecyl sulfate Cooperative Investigation, United States Department of Agriculture and Institute of Food and Agricultural Sciences, University of Florida, Florida Agricultural Experiment Station Journal Series No. 2453. Mention of a trademark, proprietary, product, or vendor does not constitute a guarantee or warrantly of the product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products or vendors that may also be suitable     

Autologous tumor vaccines processed to express α-gal epitopes: a practical approach to immunotherapy in cancer

This review describes a method by which the human natural anti-Gal antibody can be exploited as an endogenous adjuvant for targeting autologous tumor vaccines to antigen-presenting cells (APCs). Tumor cells remaining in the patient after completion of surgery, radiation, and chemotherapy are the cause of tumor relapse. These residual tumor cells can not be detected by imaging, but their destruction may be feasible by active immunotherapy. Since specific tumor-associated antigens (TAAs) have not been identified for the majority of cancers, irradiated autologous tumor vaccines have been considered as an immunotherapy treatment that may elicit an immune response against the residual tumor cells expressing TAAs. However, tumor cells evolve in cancer patients in a stealthy way, i.e., they are not detected by APCs, even in the form of vaccine. Effective targeting of tumor vaccines for uptake by APCs is a prerequisite for eliciting an effective immune response which requires transport of the vaccine by APCs from the vaccination site to the draining lymph nodes. In the lymph nodes, the APCs transporting the vaccine process and present peptides, including the autologous TAA peptides for activation of the tumor-specific T cells. The required targeting of vaccines to APCs is feasible in humans by the use of anti-Gal. This antibody interacts specifically with the -gal epitope (Gal1-3Gal1-4GlcNAc-R) and is the only known natural IgG antibody to be present in large amounts in all humans who are not severely immunocompromised. The -gal epitope can be synthesized on any type of human tumor cell by the use of recombinant 1,3galactosyltransferase (1,3GT). Solid tumors obtained from surgery are homogenized and their membranes subjected to -gal epitope synthesis. Similarly, -gal epitopes can be synthesized on intact tumor cells from hematological malignancies. Administration of irradiated autologous tumor vaccines processed to express -gal epitopes results in in situ opsonization of the vaccinating cells or cell membranes due to anti-Gal binding to these epitopes. The bound antibody serves to target the autologous tumor vaccine to APCs because the Fc portion of the antibody interacts with Fc receptors on APCs. Since patients receive their own TAAs, the vaccine is customized for autologous TAAs in the individual patient. The repeated vaccination with such autologous tumor vaccines provides the immune system of each patient with an additional opportunity to be effectively activated by the autologous TAAs. In some of the immunized patients this activation may be potent enough to induce an immune-mediated eradication of the residual tumor cells expressing these TAAs.Abbreviations Ab Antibody - Ag Antigen - APC Antigen-presenting cell - DC Dendritic cell - FcR Fc receptor - -gal epitope Gal1-3Gal1-4GlcNAc-R - 1,3GT -1,3-Galactosyltransferase - KO mice Knockout mice for 1,3GT - OVA Ovalbumin - SA Sialic acid - TAA Tumor-associated antigen     

Ethanol and amylase production by a newly isolated Clostridium sp.

A newly isolated, anaerobic, mesophillic bacterium, Clostridium sp. strain YK-3, ferments pentoses, hexoses, oligosaccharides and polysaccharides, such as soluble starch and glycogen, to ethanol and acetate. The potential of this strain for ethanol and amylase production has been examined. Ethanol was the major product and acetate a minor one. The organism could grow with soluble starch in the presence of 40 g ethanol/l. Extracellular -amylase activity was detected when the strain was cultivated with soluble starch, glycogen or dextrin. The optimum pH of this amylase was 5.5 to 7.5 with an optimum temperature of 50°C.The authors are with the Laboratory of Applied Microbiology, Faculty of Agriculture, Yamagata University, Tsuruoka 997, Japan.     

The effect of spectral light deprivation on the spectral sensitivity of the honey bee

Summary The spectral sensitivity of spontaneous phototactic behavior was tested in short wavelength deprived bees and in control bees kept outdoors. Tests were performed with a y-maze with one branch illuminated, the other dark. The relative sensitivities for the control group were: green to blue to UV = 10.272.14. Short wavelength deprived bees show a distinct decrease in their sensitivity to short wavelengths. The relation green to blue to UV here was 10.220.27. Forager bees from outdooors, short wavelength deprived for 10 days, showed a relation of green to blue to UV of 10.261.04. Electrophysiological investigation of the compound eye using electroretinogram recordings showed no difference between deprived and control animals.     

Syntheses of α- and β-Glycosyl Donors with a Disaccharide β-D-Gal-(1→3)-D-GalNAc Backbone

The synthesis of thioglycoside glycosyl donors with a disaccharide -D-Gal-(1 3)-D-GalNAc backbone was studied using the glycosylation of a series of suitably protected 3-monohydroxy- and 3,4-dihydroxyderivatives of phenyl 2-azido-2-deoxy-1-thio-- and 1-thio--D-galactopyranosides by galactosyl bromide, fluoride, and trichloroacetimidate. In the reaction with the monohydroxylated glycosyl acceptor, the process of intermolecular transfer of thiophenyl group from the glycosyl acceptor onto the cation formed from the molecule of glycosyl donor dominated. When glycosylating 3,4-diol under the same conditions, the product of the thiophenyl group transfer dominated or the undesired (1 4), rather than (1 3)-linked, disaccharide product formed. The aglycon transfer was excluded when 4-nitrophenylthio group was substituted for phenylthio group in the galactosyl acceptor molecule. This led to the target disaccharide, 4-nitrophenyl 2-azido-4,6-O-benzylidene-2-deoxy-3-O-(2,3,4,6-tetra-O-acetyl--D-galactopyranosyl)-1-thio--D-galactopyranoside, in 57% yield. This disaccharide product bears nonparticipating azido group in position 2 of galactosamine and can hence be used to form -glycoside bond. Azido group and the aglycon nitro group were simultaneously reduced in this product and then trichloroacetylated, which led to the -glycosyl donor, 4-trichloroacetamidophenyl 4,6-di-O-acetyl-2-deoxy-3-O-(2,3,4,6-tetra-O-acetyl--D-galactopyranosyl)-1-thio-2-trichloroacetamido--D-galactopyranoside, in 62% yield. The resulting glycosyl donor was used in the synthesis of tetrasaccharide asialo-GM₁.     

DNA polymorphisms associated with a new α1-antitrypsin PI Q0 variant (PI Q0riedenburg)

Summary A new PI Q0 variant (PI Q0_riedenburg) is described; it is caused by a complete deletion of the ₁-antitrypsin (₁AT) gene. The deletion gives rise to four new restriction fragment length polymorphisms (RFLPs) detected with a genomic probe of the 5 region of the gene. Analysis of the RFLPs indicates that the deletion starts immediately upstream of exon Ic. The deletion extends into the 3 flanking region of the gene but does not include the ₁AT-related gene (the PIL gene), which is located 12 kb downstream of the ₁AT gene.     

Comparison of the effects of ACE inhibition with those of Angiotensin II receptor antagonism on systolic and diastolic myocardial stunning in isolated rabbit heart

The aim was to determine whether enalaprilat (0.08 mg/kg/min) or losartan (0.01 mg/kg/min) administration before ischemia can improve postischemic systolic and diastolic dysfunction ('stunned myocardium') and attenuate the hyperfunction phase at the beginning of reperfusion. An isolated isovolumic rabbit heart preparation was subjected to 15 min of ischemia followed by 30 min of reperfusion without (group 1) or with pretreatment with enalaprilat (group 2) or losartan (group 3). Left ventricular developed pressure and end-diastolic pressure (diastolic stiffness) were measured and the time constant of isovolumic relaxation (t, Tau) and the ratio between +dP/dt and –dP/dt were calculated. In comparison to the stunned group (group 1) both enalaprilat (group 2) and losartan (group 3) exerted a significant protective effect on postischemic recovery of contractile state and diastolic stiffness. Only enalaprilat attenuated the hypercontractile phase. However, both enalaprilat and losartan failed to improve myocardial relaxation. In summary, these data strongly suggest a direct deleterious action of the local renin- angiotensin system on ischemic myocardium and diminution of myocardial stunning with its successful blockade. Although, we can not exclude the possibility that bradykinin has some cardioprotective effect, these data suggest that angiotensin exacerbates myocardial injury.