Initiation and development of wound tissue and roots on hypocotyl cuttings of Pinus sylvestris in vitro

The rooting of hypocotyl cuttings from 20-day-old seedlings of Pinus sylvestris L. cultured in vitro is discussed. About 40% of the cuttings cultured on medium lacking activated charcoal produced roots during the first two months. When activated charcoal was added to the medium, either root formation (75% formed roots) or wound tissue growth (95% formed large wound tissues) was stimulated in different experiments. These large wound tissues did not develop any roots. The anatomical changes in the basal part of the cuttings were similar during the first two weeks in all the cuttings studied. A vascular cylinder composed of short tracheids with many pores developed. Thereafter the differentiation process became varied. The amount of wound tissue produced and the time for rooting differed among the cuttings. Tracheid nests which were in contact with the vascular system in the hypocotyl via short tracheids were observed after three weeks. Subsequently, roots developed from the tracheid nests. The longer root formation was delayed, the larger the wound tissue became.
Short tracheids were found close to the wound tissue surface. Their ability to adsorb nutrients and water is discussed.     

Effects of hyperthermia on DNA interstrand crosslinking after treatment with BCNU in 9L rat brain tumor cells

The effects of hyperthermia (42 degrees C) on 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU)-mediated DNA interstrand crosslink formation were investigated in 9L rat brain tumor cells using the technique of alkaline elution. When cells were treated with 60 microM BCNU for 1 hr at 37 degrees C and incubated for 6 hr in drug-free medium at 42 degrees C, there was a 50% increase in crosslinking; and when cells were treated at 42 degrees C and incubated at 37 degrees C, there was a 45% increase in crosslinking compared with the results for cells treated and incubated at 37 degrees C. When cells were treated and incubated at 42 degrees C, there was a 129% increase in DNA crosslinking. The same relative order of results was found for cell survival. These results suggest that hyperthermia can increase DNA interstrand crosslink formation and the consequent cell death through two independent mechanisms: an increase in the amount of initial alkylation because of the increased rate of hydrolysis of BCNU at higher temperatures, and the effect of heat on DNA structure that leads to an increase in the number of crosslinks formed.     

Transient enhancement of serum somatomedin levels prior to weaning of growth-impaired vasopressin-deficient Brattleboro rats

Somatomedin serum levels of congenitally vasopressin-deficient Brattleboro rats were determined postnatally between day 1 and 55, and compared with heterozygous control values. Assays were performed with a radioimmunoassay of insulin-like growth factor 1 (IGF-1). A transient enhancement of immunoreactive IGF-1 levels between day 8 and 21 of age and a reduction in adulthood was found. This observation shows that the early growth impairments of the Brattleboro mutant are not due to a deficiency of IGF-1.     

3H-uridine labelling patterns and chromosomal polymorphism inDrosophila subobscura: J and U chromosomes

The³H-uridine labelling patterns in J and U polytene chromosomes ofDrosophila subobscura were determined. The analysis was carried out in two developmental stages and in two strains proceeding from the same geographical origin whose genotypes were: J_st/J_st; U₁₊₂/U₁₊₂ and J₁/J₁; U₁₊₂₊₈/U₁₊₂₊₈ respectively. It was observed that the labelling pattern coincided very approximately with the puffing pattern in the same stages and chromosomal arrangements. Comparison of the³H-Uridine incorporation patterns between chromosomal arrangements showed light quantitative differences. These results are discussed in relation to the inversion effect.     

The thermodynamics of complex formation of cyclic tetra-aza-tetracetic acids

Enthalpy changes for the complexation of alkaline-earth and transition metals with three cyclic tetra-aza-tetracetic acids (cDOTA, cTRITA and cTETA) were obtained by continuous titration calorimetry. From these values and free energy data, the entropy changes for the same reactions were derived. The results show that these complexes are stabilised by both favourable enthalpy and entropy changes, except those of Mg²⁺ and those of Sr²⁺ and Ba²⁺ with cTETA. Generally, the entropy changes for the reactions of the alkaline-earth metals are higher than for the reactions of the non cyclic polyaminocarboxylic acids, but for the reactions of the transition metals the entropy changes are comparable for the cyclic and non cyclic ligands. These results are discussed in terms of a model of ‘cage’ coordination of the metals.The enthalpy changes decrease with the increase in size of the tetra-aza ring (except in the case of Cu²⁺) but no specific cavity size effect is noticeable. Consideration of the temperature-dependent and temperature-independent contributions to ΔH supports the idea that the number of coordinated nitrogen atoms and carboxylate groups vary along the series.     

High density of transmembrane glycoproteins on the flagellar surface of boar sperm cells

Membrane halves of boar sperm flagella were produced by freeze-fracture and labeled in situ with concanavalin A and wheat germ agglutinin; the lectins were visualized with protein-gold complexes. Concanavalin A and wheat germ agglutinin binding sites partition with both protoplasmic and exoplasmic halves of the membrane. A high density of lectin marking was found on protoplasmic membrane halves; we conclude that the label corresponds to transmembrane glycoproteins that, on freeze-fracture, are dragged across the outer (exoplasmic) half of the phospholipid bilayer. Our demonstration of numerous transmembrane proteins in sperm flagella offers the structural setting for previous models on flagellar surface motility that postulate accessibility of motile membrane components to the submembranous cytoskeleton.     

Ovarian response to active immunization against luteinizing hormone in the cow

Two experiments were conducted to determine whether active immunization against luteinizing hormone (LH) could lead to ovarian cyst development in the cow. In Experiment 1, cyclic beef heifers were randomly assigned to receive bovine LH (bLH) conjugated with ovalbumin (LH-immunized; n=4) or ovalbumin alone (control; n=5). Blood samples were collected at monthly intervals from the LH-immunized heifers to determine antibody titers. Heifers were observed for estrous behavior twice daily. All heifers were slaughtered 4 mo after initial immunization and ovaries examined for follicular status. In Experiment 2, mature dairy cows were immunized with bLH (LH-immunized; n=4) or ovalbumin alone (control; n=3). Weekly blood samples were collected from all cows for 26 wk and ovaries were rectally palpated. Sera from all of the LH-immunized heifers and cows had antibodies to LH. All of the LH-immunized animals stopped cycling 1 mo after immunization. In spite of the fact that serum follicle stimulating hormone levels were unaffected, ovarian cysts could not be found in either the LH-immunized heifers or cows.     

Possibility of reducing the luteolytic dose of cloprostenol in cyclic dairy cows

The administration of cloprostenol by intravulvosubmucous (i.v.s.m.) injection at 1 2 and 1 4 of the dose usually given by intramuscular (i.m.) injection, was tested in dairy cows for luteolysis and estrus synchronization. The i.m. injection was used in ten adult cows at the usual dose of 500 mug/animal. Eleven adult cows and 11 heifers were treated i.v.s.m. with a dose equivalent to 250 mug/animal and 125 mug/animal, respectively. Two injections of cloprostenol were administered 11 days apart to the cows not detected in oestrus after a single injection. Forty-three out of the total 46 animals were detected to be in dioestrus at the time of at least one of the injections, as reflected by the plasma progesterone concentrations at the time of treatments. Three out of the 43 animals injected during dioestrus were refractory to the luteolytic effect of cloprostenol; this appeared to be independent of the dosage and the route of administration (refractory cows were: one adult cow treated i.m. and two treated i.v.s.m. with 125 mug of cloprostenol). The mean time interval from injection to the onset of heat was 82.8 hours with a confidence limit for 95% of probability between 67.9 hours and 92.7 hours. The difference between treatments is not significant. The results suggest that in heifers and adult cows cloprostenol can be given i.v.s.m. route at a reduced dose of 1 4 of the usual 500 mug i.m. dosage without affecting the luteolytic effect of the drug or fertility.     

Antibodies to spiroperidol and their anti-idiotypes as probes for studying dopamine receptors

Spiroperidol was covalently conjugated to bovine serum albumin (BSA). Conjugated spiroperidol was almost as efficient as free spiroperidol in its binding capacity to dopamine receptor. Antibodies to spiroperidol were produced in rabbits following repeated immunizations with the conjugate of spiroperidol and BSA. The obtained antibodies have an apparent K_D of 0.02 nM for [³H]-spiroperidol. These antibodies bind also to other butyrophenones with IC₅₀ values three to four orders of magnitude higher than the IC₅₀ obtained with unlabeled spiroperidol. Antibodies were purified from anti-spiroperidol sera by affinity chromatography. Anti-idiotypic antibodies were raised in rabbits by immunization with the purified anti-spiroperidol antibodies. Some rabbits produced anti-idiotypic antibodies which bind to rat and calf striatum.