Current status of erythrocyte substitutes.

During the last two decades the search for alternatives to whole blood transfusions has led to promising developments in the field of erythrocyte substitutes. Hemoglobin solutions free of fragments of erythrocyte stroma and fluorocarbon emulsions are not blood-type-specific and appear likely to satisfy some proportion of our blood requirements. Both must be modified before becoming clinically useful. The oxygen affinity of the hemoglobin solution must be reduced and its intravascular persistence improved. Fluorocarbons cannot yet contribute significantly to the oxygen supply unless the patient breathes hyperbaric oxygen. Recent advances are leading to solutions for these problems.     

Occurrence of aflatoxins in oilseeds providing cocoa-butter substitutes.

Four oilseeds providing cocoa-butter substitutes--shea, pentadecima, illipe, and salseed--when tested as substrates for aflatoxin production by two strains of Aspergillus parasiticus, gave varying levels of aflatoxin. Aflatoxins were found at low levels occurring naturally in moldy shea-nuts, but none of 21 commercial shea-nut samples contained greater than 20 micrograms of aflatoxin B1 per kg.     

Ultrasonic baths as substitutes for shaking incubator baths.

An easily assembled incubation bath for enzyme work is described. The bath is made from commercially available units: an ultrasonic bath of the type used for cleaning and a temperature-controlling device. The device is not only much cheaper, quieter, and more compact than a commercially built shaking-type bath, but is also gives superior mixing of heterogeneous enzyme incubation samples, particularly those containing tissue homogenates or subcellular particles.     

Sugar sensing in higher plants.

Sugar repression of photosynthetic genes is likely a central control mechanism mediating energy homeostasis in a wide range of algae and higher plants. It overrides light activation and is coupled to developmental and environmental regulations. How sugar signals are sensed and transduced to the nucleus remains unclear. To elucidate sugar-sensing mechanisms, we monitored the effects of a variety of sugars, glucose analogs, and metabolic intermediates on photosynthetic fusion genes in a sensitive and versatile maize protoplast transient expression system. The results show that sugars that are the substrates of hexokinase (HK) cause repression at a low concentration (1 to 10 mM), indicating a low degree of specificity and the irrelevance of osmotic change. Studies with various glucose analogs suggest that glucose transport across the plasma membrane is necessary but not sufficient to trigger repression, whereas subsequent phosphorylation by HK may be required. The effectiveness of 2-deoxyglucose, a nonmetabolizable glucose analog, and the ineffectiveness of various metabolic intermediates in eliciting repression eliminate the involvement of glycolysis and other metabolic pathways. Replenishing intracellular phosphate and ATP diminished by hexoses does not overcome repression. Because mannoheptulose, a specific HK inhibitor, blocks the severe repression triggered by 2-deoxyglucose and yet the phosphorylated products per se do not act as repression signals, we propose that HK may have dual functions and may act as a key sensor and signal transmitter of sugar repression in higher plants.     

Albuminated dacron protheses as improved blood vessel substitutes.

An improved vascular graft was achieved by treating knitted Dacron prostheses with cross-linked albumin. Eight segments, each six centimeters in length, were coated and implanted in the canine thoracic aorta for periods ranging from 24 hours to six months. The results were compared with the eight untreated grafts implanted for similar periods as control experiments. Albumin coating resulted in a faster healing : cells appeared earlier and collagen development was greater. The most beneficial effect of that treatment was the reduction in embolization, as shown by the considerable decrease in kidney damage.     

Rheological aspects of mucin-containing solutions and saliva substitutes.

In this study rheological properties of aqueous solutions of mucin, albumin and mucin-albumin have been investigated in search for saliva substitutes. They were compared with commercially available saliva substitutes on the one hand and natural human saliva on the other hand. For the latter a few measurements on saliva are reported here in addition to previously reported measurements done in our laboratory. Proteins absorb at the interface and saliva proteins do so strongly and rapidly. Therefore rheological measurements were carried out on the interface and on the bulk underneath the layer. In both cases the flow curve and the complex viscosity was determined. The results show that specific mucin-albumin solutions were rheologically similar to human whole saliva with respect to both bulk liquid and surface properties. The rheological properties of commercial saliva substitutes were essentially different from those of human saliva. It is concluded that mucin-albumin solutions have good perspectives as saliva substitutes.     

Recombinant haemoglobin in the development of red-blood-cell substitutes.

Increasing concern over viral contamination of blood is spurring the development of a blood substitute which can effectively replace the oxygen-carrying capabilities of transfused erythrocytes. Solutions of chemically modified haemoglobin represent one option being evaluated for this role. More recently, recombinant-DNA techniques have enabled production of human haemoglobin in host expression systems, and progress is being made towards the creation of a genetically engineered molecule incorporating the properties required of a blood substitute.     

Sugar transporters in plant biology.

Sugar transporters are key players in many fundamental processes in plant growth and development. Recent results have identified several new transporters that contribute to a wide array of physiological activities, and detailed molecular analysis has provided exciting insights into the structure and regulation of these essential membrane proteins.     

Sugar transport in Coprinus cinereus.

Two transport systems for glucose were detected: a high affinity system with a Km of 27 muM, and a low affinity system with a Km of 3.3 mM. The high affinity system transported glucose, 2-deoxy-D-glucose (Km = 26 muM), 3-O-methylglucose (Km = 19 muM), D-glucosamine (Km = 652 muM), D-fructose (Km = 2.3 mM) and L-sorbose (Km = 2.2 mM). All sugars were accumulated against concentration gradients. The high affinity system was strongly or completely inhibited by N-ethylmaleimide, quercetin, 2,4-dinitrophenol and sodium azide. The system had a distinct pH optimum (7.4) and optimum temperature (45 degrees C). The low affinity system transported glucose, 2-deoxy-D-glucose (Km = 7.5 mM), and 3-O-methylglucose (Km = 1.5 mM). Accumulation again occurred against a concentration gradient. The low affinity system was inhibited by N-ethylmaleimide, quercetin and 2,4-dinitrophenol, but not by sodium azide. The rate of uptake by the low affinity system was constant over a wide temperature range (30--50 degrees C) and was not much affected by pH; but as the pH of the medium was altered from 4.5 to 8.9 a co-ordinated increase in affinity for 2-deoxy-D-glucose (from 52.1 mM to 0.3 mM) and decrease in maximum velocity (by a factor of five) occurred. Both uptake systems were present insporelings germinated in media containing sodium acetate as sole carbon source. Only the low affinity system could initially be demonstrated in glucose-grown tissue, although the high affinity system was restored by starvation inglucose-free medium. The half-ti me for restoration of high affinity activity was 3.5 min and the process was unaffected by cycloheximide. Addition of glucose to an acetate-grown culture inactivated the high affinity system with a half-life of 5--7.5 s. Addition of cycloheximide to an acetate-grown culture caused decay of the high affinity system with a half-life of 80 min. Regulation is thus thought to depend on modulation of protein activity rather than synthesis, and the kinetics of glucose, 2-deoxy-D-glucose and 3-O-methylglucose uptake would be consistent with there being a single carrier showing negative co-operativity. Analysis of transport defective mutants revealed defects in both transport systems although the mutants used were alleles of a single gene. It is concluded that this gene (the ftr cistron) is the structural gene for an allosteric molecule which serves both transport systems.     

Fluorocarbons as blood substitutes. Toxicity in the rat]

Liquid fluorocarbons, having high solubility for gases (O2, CO2...) have been used as artificial blood substitutes in animals with variable results. The great diversity of these products and the lack of reproductiveness in their composition have not permitted, up to now, a standardization of their utilisation norms. Our work was to study the toxicity, in the rat, of a new kind of fluorocarbon emulsion (E-66, Ugine-Kuhlmann, France) used as an artificial blood substitute during exchange-perfusion. The short survival of the rats is in opposition to the good in vitro results obtained in other experiments (high solubilities of oxygen and dioxide carbon). The toxicity of this fluorinated emulsion is demonstrated by histologic lesions in lungs, liver and kidneys and the great amount of fluor stored in these organs. The mechanism of this toxicity is still to be demonstrated. Hepatic lesions (hyperhemia with dilatation of central veins and sinusoids) and pulmonary lesions (vascular congestion, alveolar oedema) prove a circulatory disturbance leading to right-sided cardiac failure. However, cellular degenerescence lesions, observed in hepatocytes and renal tubular cells, do not permit to exclude formally a cellular toxicity of the E-66 emulsion.     

Biological substitutes for pesticides

In the 20th century an increasing number of pesticides, based on biocidal molecules, were the means for a substantial increase in food and fibre production and quality. Because of health and environmental concerns continued extensive use of such molecules is intensively debated and substitutes are often urgently required. Beside crop plant resistance, various biological control methods based on natural pest suppressing organisms are regarded as main alternatives. Several approaches and concepts also have been tested and commercial organism-based preparations are steadily increasing. However, further biotechnological efforts are required to give them status of being practical substitutes to pesticides. At present they are not comparable to pesticides in meeting efficacy, market and other expectations, but they still have a promising future, especially where genetically modified organisms can be used.