The use of an optical brightener in the study of plant structure.

An optical brightener Calcoflour White M2R New has been used to stain cell walls of higher plants. It can be used either as a vital stain for intact plants or for hand sections and plastic-embedded thin sections. Walls are brilliantly fluorescent while most cytoplasmic components are normally unstained. The brightener binds strongly to cellulose, carboxylated polysaccharides, and callose. Staining for 20 sec to 2 min in a 0.01% solution of the brightener is preferred for most purposes.     

Conversion of methionine to homocysteine thiolactone in liver

Since hemocysteinemia is associated with arteriosclerosis, the conversion of methione to homocysteine thiolactone was studied in guinea pig liver in vivo. 60 min after intraperitoneal injection of [¹⁴C]methione, [¹⁴C]homocystein thiolactone was found to constitute 9.1% ± 0.2 of the lipid bound ¹⁴C and 20% ± 1.0 of the acid soluble ¹⁴C. This conversion is the first step of a new pathway by which the sulfur of methionine is transferred to phosphoadenosine phosphosulfate.     

Characteristics of lengthening contractions associated with injury to skeletal muscle fibers

Lengthening (eccentric) contractions result in injury to skeletal muscle fibers. Two hypotheses were tested through lengthening contractions of an in situ muscle preparation: the extent of injury increases with increases in the duration; and the extent of injury increases with increases in the peak force. Mice were anesthetized, and distal tendons of the extensor digitorum longus muscles were attached to a servomotor. Muscles were stimulated at 150 Hz and lengthened 20% of fiber length (Lf). Lengthening contractions were performed at 0.2, 0.5, or 1.0 Lf/s with durations of 0.5-15 min. Peak force during lengthening contractions at 1.0 Lf/s was decreased by inducing fatigue with isometric contractions, stimulating at 70-100 Hz, or 3) lengthening 10% of Lf. Injury was assessed 3 days after lengthening contractions by histological appearance and maximum force (Po) development. Injury increased with duration up to 5 min. After 5 min, fatigue appeared to prevent further injury. Results for 0.2 and 0.5 Lf/s were similar to those for 1.0 Lf/s but with less injury. A high correlation was observed between histological appearance of injury and the decrease in Po. The extent of injury was related to the peak force developed during the lengthening contractions.     

Muscle metabolism in track athletes, using 31P magnetic resonance spectroscopy.

We tested whether preferred running event in track athletes would correlate with the initial rate of phosphocreatine (PCr) resynthesis following submaximal exercise. PCr recovery was measured in the calf muscles of 16 male track athletes and 7 male control subjects following 5 min of repeated plantar flexion against resistance. Pi, PCr, and pH were measured using phosphorus magnetic resonance spectroscopy (31P MRS) with an 8-cm surface coil in a 1.8-T magnet. During exercise, work levels were gradually increased to deplete PCr to 50-60% of the initial value. No drop in pH was seen in any of the subjects during this exercise. The areas of the PCr peaks following exercise were fit to monoexponential curves. Two or three tests were performed on each subject and the results averaged. Athletes were divided into three groups based on their primary event: sprinters running 400 m or less, middle-distance athletes running 400-1500 m, and long-distance athletes running farther than 1500 m. The maximal rates of PCr resynthesis (mmol.min-1.kg-1 muscle weight) were 64.8 +/- 8.6, for long-distance runners; 41.4 +/- 11, for middle-distance runners; 32.0 +/- 7.0, for sprinters; and 38.6 +/- 10, for controls (mean +/- SE). The faster PCr recovery rates seen in long-distance runners compared with sprinters indicate greater oxidative capacity, which is consistent with the known differences between athletes in these events.     

Leaf teeth, transpiration and the retrieval of apoplastic solutes in balsam poplar

The rapid flow of the transpiration stream through major veins to leaf teeth was followed in leaves of Populus balsamifera L., using the tracer sulphorhodamine G (SR), which probes for cells with H⁺-extrusion pumps. The tracer accumulated quickly in the hydathodes of the teeth. It was shown by freeze-substitution and anhydrous processing that SR was taken up by phloem parenchyma and epithem cells of the hydathode. When ¹⁴C-labelled aspartate was fed to the leaves in the transpiration stream, it also was taken up most strongly by the same phloem parenchyma and epithem cells. It is proposed that one function of the hydathodes in leaf teeth is the retrieval of solutes from the transpiration stream.     

Diet composition of starry smooth-hound Mustelus asterias and methodological considerations for assessing the trophic level of predatory fish

The stomach contents of 640 starry smooth-hound Mustelus asterias from the north-east Atlantic were examined. The diet was dominated by crustaceans (98.8% percentage of index of relative importance, %IRI), with the two main prey species being hermit crab Pagurus bernhardus (34% IRI) and flying crab Liocarcinus holsatus (15% IRI). Ontogenetic dietary preferences showed that smaller individuals [20–69 cm total length (L_T) n = 283] had a significantly lower diversity of prey than larger individuals (70–124 cm L_T, n = 348); however, 18 prey species were found exclusively in smaller individuals and eight prey taxa were found exclusively in larger individuals. Larger commercially important brachyurans such as edible crab Cancer pagurus and velvet swimming crab Necora puber were more prevalent in the diet of larger individuals. Specimens from the North Sea ecoregion had a lower diversity of prey types for a given sample size than fish from the Celtic Seas ecoregion. Whilst cumulative prey curves did not reach an asymptote, this was primarily due to the high taxonomic resolution utilized and 95% of the diet was described by just seven crustacean taxa. The trophic level (TL) was calculated as 4.34 when species-level prey categories were used. This fine-scale taxonomic resolution resulted in a TL estimate close to a whole level above that estimated using wider taxonomic groupings. This large bias has important methodological implications for TL studies based on categorized prey data, particularly those of predatory fish.     

Proton pump activity in bundle sheath tissues of broad-leaved trees in relation to leaf age

The fluorescent probe sulphorhodamine G (SR) has been previously used as an indicator of low extra-cellular pH and, by inference, of proton extrusion activity in living leaves. In legumes the SR uptake and proton extrusion was characteristic of the extended bundle sheath system (EBS) or paraveinal mesophyll, composed of bundle sheath cells and the related network of bridging cells between veins. This system has been identified as a site of temporary storage of amino carbon in soybean. A tree species. Populus deltoides Bartr. ex Marsh, was known both to have the EBS system in its leaves and to carry organic nitrogen in its xylem sap. It is now shown that P. deltoides also accumulates the SR probe in the EBS system. This association has been explored in 8 other broad-leaved tree species. Seven of the 8 species have EBS systems and accumulate SR in them in early summer. The 8th species, Tilia americana L. has no EBS system and shows weak SR accumulation. The capacity to accumulate SR (and by inference to scavenge solutes from the transpiration stream) disappeared in all species at various stages in late summer. In two species, in addition, SR accumulation is interrupted for several weeks during fruit growth. It is proposed that EBS systems will be found in many dicotyledonous leaves, and will be found to scavenge solutes, especially organic nitrogen, from the xylem sap.     

Construction of cosmid genomic libraries for the normal and myopathic Syrian hamsters

Cosmid genomic libraries from both normal and myopathic Syrian hamsters have been constructed. MboI was used to generate 35- to 50-kilobase DNA fragments which were isolated from a 5-25% NaCl gradient. The 35- to 50-kilobase DNA fragments were ligated to the cosmid vector pCV108 and packaged into Escherichia coli DK1. Approximately 3 X 10(5) - 4 X 10(5) clones were obtained per microgram of ligated DNA. Thirteen clones have been isolated from 2 X 10(5) colonies using a cardiac myosin heavy chain clone as a probe. Restriction maps of two of these clones are presented here.