Investigations on circumferential microfilament bundles in rat retinal pigment epithelium

The presence of contractile proteins in normal rat retinal pigment epithelium has been studied using fluorescence and electron microscopy. Investigations using the F-actin binding toxin, phallacidin, coupled to the fluorochrome nitrobenzoxadiazole, revealed a band of fluorescence at or near the cell membrane. Immunofluorescent observations with anti-myosin and anti-alpha-actinin antisera gave similar results. Electron microscopy employing glutaraldehyde-8% tannic acid fixation revealed the presence of a circumferential microfilament band beneath the pigment epithelial apical surface that is closely associated with the plasma membrane and junctional complexes. Freeze-fracture studies confirmed the relationship of this band to the junctional complexes. The microfilament band measures approximately 0.5 micron +/- 0.2 micron in width and is composed of numerous 6 to 7 nm filaments. Some microtubules are seen in regions around the band, but no organelles appear to be associated with this structure. In en face sections through the zonula adherens, the circumferential microfilament band is associated with 30-nm electron-dense particles that are bound to the internal side of the membrane. Morphological evidence suggests that these may serve in anchoring the band to the membrane and assist in aligning the microfilament bands of adjoining cells. In the subapical cytoplasm, a microfilament bundle network was detected that interfaced with the circumferential microfilament band. In some cases, pigment epithelium was incubated in media-199 containing 25 to 50 ng/ml phallacidin prior to fixation. Circumferential microfilament bands of tissues treated in this manner exhibited a striated appearance.     

Temperature regulation in a burrow-nesting tropical seabird,the Wedge-tailed Shearwater (Puffinus pacificus)

Summary At low air temperatures (2.3–13.9°C), Wedge-tailed Shearwaters (Puffinus pacificus) shivered and their oxygen consumption increased to as much as 283% of the mean value (0.77 ml O₂/g·h) within the thermoneutral zone of air temperature (23–34°C). The minimal thermal conductance of the tissues and plumage was similar to the value predicted from the body mass (320.5 g). The oxygen consumption of the birds within their thermoneutral zone was lower than predictions based on body mass. At elevated air temperatures, the shearwaters panted at respiratory frequencies as high as 260 respirations/min; maximal respiratory frequencies were not invoked until the birds had become hyperthermic. During exposure to a hot environment, the oxygen consumption of the birds increased and in most instances the shearwaters were not able to lose heat equivalent to their concurrent metabolic heat production.Symbols and abbreviations BMR basal metabolic rate - C _total total thermal conductance - f respiratory frequency - TEWL total evaporative water loss - T _st stomach temperature - T _re rectal temperature     

Genetic study on indole-3-acetic acid production by ectomycorrhizal Hebeloma species: inter- and intraspecific variability in homo- and dikaryotic mycelia

Summary Interspecific variability of indole-3-acetic acid (IAA)-synthesizing activity was examined within 12 wild strains of different Hebeloma species. Interstrain variability was studied within 11 wild strains of Hebeloma cylindrosporum (Romagnési) and intrastrain variability was considered by using 20 homokaryotic and 50 controlled dikaryotic mycelia belonging to the progeny of one laboratory fruiting strain of this species.The range of variation of IAA-synthesizing activity was of the same order of magnitude within the four groups considered. No correlation was detected between, on one hand, the IAA-synthesizing activity of the mycelia and, on the other hand, their taxonomic position, their geographic origin, or their host plant.Within the progeny of one H. cylindrosporum fruiting strain, 15 of the 50 controlled dikaryons presented an activity higher than that of the original dikaryon. The variation among dikaryons could not be strictly related to the variation in parental homokaryons, indicating that genetic control of this activity probably involves a nonadditive component. Significant additive and nonadditive components of the genetic variation were detected, each of them representing about 50% of the total variation. The nonadditive heritable component could not be explained by a model involving only dominance.     

The association between spontaneous pyelonephritis and maturity-onset diabetes mellitus in male MM mice

Blood glucose and glucose tolerance tests demonstrated that many male MM mice are diabetic. Serial urine sampling showed that the diabetes occurred only in mature MM males and consisted of a single self-limiting episode. Histological examination of the pancreas, together with measurements of body weight, glycosylated haemoglobin and plasma insulin, revealed that the diabetes was of the maturity-onset insulin-resistant type. Bacteriological examination of the urine samples showed that urinary tract infection, a known feature of male MM mice, occurred in the diabetics but only after the onset of hyperglucosuria. It was concluded that the high urinary glucose levels of diabetic MM males are of prime importance in the aetiology of the renal infection which occurs rarely in non-diabetic MM males or in other strains in the colony. An infectious aetiology for the diabetes per se was excluded by the existence of diabetes in germfree MM males.     

Antitrypanosomal action of cis-diamminedichloroplatinum (II) analogs

The present report deals with the alterations produced by cis-diamminedichloroplatinum (II) (DDP), and 2 of its analogs: cis-Pt(II)(tranylcypromine)2Cl2 and cis-Pt(II)(benzothiazole)2Cl2 in cultured epimastigote forms of Trypanosoma cruzi. Studies have been performed at the ultrastructural level and the inhibitory effect of these complexes on macromolecule synthesis, evaluated by 3H-thymidine, 3H-uridine, and 3H-leucine incorporation, has been investigated. DDP at concentrations of 50 and 100 micrograms/ml does not inhibit significantly the incorporation of radioactive precursors, but a clear decrease was observed with the 2 analogs. Eight hours of treatment at a concentration of 10 micrograms/ml rendered in all 3 cases an increase in autophagic vacuoles and lipids as well as an abnormal condensation of the nucleus chromatin.     

Biochemical investigation of tissue growth: towards definition of "standard" tissue samples

This paper presents an animal model [the kangaroo], a quantitative anatomical dissection procedure, and a mathematical model [two-phase linear regression] which illustrate that body tissues grow at varying rates relative to each other. An argument is developed that biochemists interested in tissue chemical activity need to be able to sample tissue of known [predicted] growth rate. It is assumed that the ability to select, say muscle tissue samples, from any one animal at a stage of its growth where the individual selected pieces of tissue have known [predicted] low, average and high growth rates would allow comparisons to be made between the sampled tissues that may elucidate the underlying biochemical mechanisms involved in the growth process. It is asserted that to establish standards for tissue samples used in biochemical growth studies, the growth rate of the sampled tissue should be one of the criteria incorporated into the definition of what is "standard" for a tissue sample.     

Alteration of the anterior acrosome of motile bovine spermatozoa by fructose and hydrogen ion concentration

Storing cauda epididymal spermatozoa in seminal plasma or in defined media at 1 x 10(9) spermatozoa/ml for 24 h at 4 degrees C caused swelling of the apical ridge on motile spermatozoa (SAR) provided concentrations of fructose in the range normally found in seminal plasma or comparable levels of glucose were present. Evaluation of these conditions indicated that, with glycolysable sugars in the media, pH dropped from 6.6-6.7 to 5.7-6.0. Most of the pH decrease occurred during the first 2 h of slow cooling from 37 to 4 degrees C. pH decrease was undoubtedly due to sperm organic acid production which overwhelmed the relatively weak buffering capacity of the defined media and/or seminal plasma. Inducing pH decreases with HCl in fructose-free conditions, and using NaOH to prevent a pH decrease when fructose was included in media, demonstrated that exposing spermatozoa to pH values of 5.7-6.0 and not a specific response to fructose was the major cause of SAR.     

Effects of prostaglandins E-2 and F-2 alpha on the metabolism of [U-14C]glucose by mouse morulae-early blastocysts in vitro

During 5-h culture in the presence of radioactive glucose, PGE-2 (10 micrograms/ml) significantly inhibited incorporation of glucose into the acid-soluble glycogen pool. PGE-2 at 1 and 10 micrograms/ml and PGF-2 alpha at 1 microgram but not 10 micrograms/ml stimulated incorporation of glucose into non-glycogen macromolecules during culture. However, the utilization of acid-soluble glycogen and other biochemical pools was not affected by the presence of PGs in the medium during 24-h chase culture of pulse-labelled embryos. Carbon dioxide production was significantly suppressed in the presence of PGs but accumulation of lactate was not affected. The results indicate that PGE-2 and PGF-2 alpha, in physiological concentrations, directly influence the metabolism of glucose by preimplantation embryos.