Epidermal proliferation characteristics are similar in the pilary canal of mouse hair follicles and in interfollicular epidermis

Proliferation characteristics of basal cells in the pilary canal of resting hair follicles were investigated and compared with corresponding parameters in interfollicular epidermis of hairless mice. The mitotic rates had similar 24-h means at both locations. Distinct circadian rhythms which showed phasing and amplitudes similar to that in interfollicular epidermis, were demonstrated by the 3H-TdR labelling index, the mitotic rate and the mitotic index. Influx of cells to and efflux of cells from the S phase were measured in the early morning and in the evening by a 3H-TdR double labelling method. The influx values were similar at both times of both locations. The efflux values recorded in the morning were more than twice the values seen in the evening in both the pilary canal and in interfollicular epidermis. The epidermal motitic rate in the pilary canal was depressed by epidermal extracts, and increased after adhesive tape stripping in the same way as in interfollicular epidermis. The results indicate no heterogeneity in cell proliferation characteristics between the two locations, and suggest that similar mechanisms are responsible for maintainance of growth equilibrium at both sites.     

Detection of circulating Dirofilaria immitis antigens in random source laboratory dogs: evaluation of two commercial serodiagnostic tests

Two commercially available serodiagnostic tests for Dirofilaria immitis antigens were evaluated for sensitivity, specificity, predictive values, and reliability using serum from 110 random source dogs. Both tests were performed in two separate laboratories on serum samples randomized in five blocks of 22 samples each. Dogs were examined for microfilariae using the modified Knott's technique, and for adult parasites by necropsy. Forty-eight of the 110 dogs (43.6%) had either adult or juvenile parasites within the cardiopulmonary vasculature or microfilariae in the peripheral blood. Of those 48, 26 (54.2%) were amicrofilaremic and had cardiopulmonary parasite populations ranging from one to greater than 50. In both laboratories, both commercial tests failed to detect infection in eight of the 26 amicrofilaremic dogs. Three amicrofilaremic dogs were positive by both tests in both laboratories. Four dogs (3.6%) had microfilariae without adults. Two of those four dogs were negative by both commercial tests in both laboratories. One commercial test had 38 false negatives in one laboratory, 13 of which were also negative in the second laboratory. The other test had 21 false negatives in one laboratory and 20 in the other laboratory. Fourteen of these samples were falsely negative in both laboratories. False positives were low in both laboratories for both tests.     

Biochemical study of cardiac valvular tissue. Biosynthesis in vitro of hexosamine-containing substances in bovine heart valve

1. Two distinct hexosamine-containing substances have been obtained from bovine cardiac valvular tissue which was incubated with labelled glucosamine. These were identified as mucopolysaccharides and glycopeptides respectively, both by the elution pattern from a Sephadex G-50 column and by chemical analysis. 2. In the mucopolysaccharide fraction about 80% of both the total hexosamine and total radioactivity were present; the galactosamine/glucosamine ratio of the amounts was about 1.36. 3. The glycopeptide fractions had about 20% of both total hexosamine and total radioactivity; the galactosamine/glucosamine ratio of amounts was about 0.44. In this fraction, over 15% of radioactivity was present in sialic acid. 4. In contrast with the concentrations of the several chemical components, there were remarkable differences in the biosynthetic activities among the four valves; the tricuspid valve had highest specific radioactivity in all components of both substances, followed then in turn by mitral, aortic and pulmonary valves. 5. Glucosamine in mucopolysaccharides had a rapid rate of turnover, followed then in turn by turnover rates of both glucosamine and galactosamine in glycopeptides, and of galactosamine in mucopolysaccharides. The possible significance of these findings is discussed.     

The cleavage of the Met-Lys bond in a bradykinin derivative by glandular kallikreins

The synthetic tridecapeptide Gly-Leu-Met-Lys-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg was used as a model substrate for horse urinary and porcine pancreatic kallikreins. The Met-Lys bond is hydrolyzed selectively by both enzymes. Oxidation of the methionine residue to sulfoxide made the peptide resistant to both kallikreins. Substitution of either the methionine or lysine residues by norleucine led to peptides in which the Nle-Lys or the Met-Nle bonds, respectively, were susceptible to the urinary kallikrein. The esterolytic and Met-Lys bond-splitting activities of both enzymes were inhibited similarly by phenylmethanesulfonyl fluoride. Both activities of the pancreatic kallikrein were inhibited by the chloromethane derivative Ala-Leu-Lys-CH2Cl. Inhibition by benzamidine of Met-Lys hydrolysis by both kallikreins was observed.     

Aggression in shorebirds in relation to flock density and composition

The aggressive interactions of Turnstones and Purple Sandpipers feeding in wintering mixed-species wader flocks could be classified into those involving food and those involving space. All observed interspecific encounters were of short duration and were initiated and won by the larger species; the majority did not involve food and were resolved by low-intensity displays. Intraspecific interactions (a greater proportion of which involved food) were also resolved quickly and were usually won by the initiator. Space-related encounters between conspecifics were more likely to be resolved than food-related encounters just by threat displays. Aggression rates increased with flock densities. However, the increase in aggression with density was dependent on the species composition of the flock: both Turnstones and Purple Sandpipers were more likely to be involved in fights (both over food and over space) with conspecifics than with other species, indicating that the aggression costs of flocking were less in mixed-species flocks.     

Changes in ribosomes associated with spore senescence in the bean rust fungus.

Studies were carried out to idenify the cause of the decline in transferase activity and capacity to bind polyuridylic acid which occurs in ribosomes from germinated uredospores of the bean rust fungus, Uromyces phaseoli (Pers.) Wint., aged longer than 6 h on a water surface. We have shown that such ribosomes lose the capacity to respond to added transferase-I and that both subunits were affected by the ageing process. These changes were not accompanied by a significant alteration in the composition of the ribosome. However, deoxycholate had a greater detergent effect on ribosomes from germinated spores than from nongerminated spores as shown both by loss of capacity to polymerize amino acids and loss of protein. Ribonuclease activity did not increase during germination, but the amount found (Imug/g spores) was easily detectable. It was suggested that loss of response to transferase-I was due to an alteration of ribosomal proteins of both subunits.     

Reproductive rates of tsetse flies in the field in Zimbabwe

Abstract. Tsetse flies Glossina morsitans morsitans Westwood and G. pallidipes Austen were marked and released within 12 h of emergence at Rekomitjie Research Station, Zambezi Valley, Zimbabwe, and on Redcliff Island, Lake Kariba. Ovarian dissections were performed on recaptured flies and on wild collected samples. At Rekomitjie >90% of female G. m. morsitans were inseminated by age 4 days and G. pallidipes by 7 days. For both species at both sites the length of the largest oocyte, for flies in ovarian category zero, increased approximately linearly for about the first 6 days and was ovulated at c. 6–8 days. The largest oocyte grew significantly more slowly in later cycles. For G. m. morsitans , but not for G. pallidipes , the rate increased with temperature; the rates were always higher than observed in the laboratory. At Rekomitjie, for both species and at a mean screen temperature of 22C, the first larva was produced at c. 18 days and subsequent larvae at 11–day intervals; the intervals decreased with temperature by c. 0.5 days/C. On Redcliff Island the intervals for both species were 2 days shorter than at Rekomitjie at any given screen temperature and were sometimes as short as 7 days. The length of the larva in utero increased exponentially during pregnancy.     

Enhanced Microbial Removal of Pyrene in Soils in the Presence of Earthworms

Microbial degradation of pyrene was studied in soils in the presence and absence of earthworms (Eisenia foetida) to demonstrate an integrated innovative strategy for bioremediation of sites lightly polluted by polycyclic aromatic hydrocarbons. Desorption of pyrene and soil microbial respiration were measured to elucidate the mechanism of enhanced microbial degradation. The results showed that both soil properties and contact time could influence pyrene biodegradation. The introduction of E. foetida enhanced pyrene removal significantly both in freshly spiked and aged soils. The percentage pyrene removal in the presence of E. foetida was 45.5–91.0% after 14 d of incubation, which were 2.1 to 2.8 times greater than those without the worms. The enhanced pyrene removal is attributed to both enhanced microbial degradation and uptake by the worms. Microbial degradation of pyrene increased by 1.2 to 1.6 times in the presence of the worms. Overall, the introduction of live worms could improve both pyrene bioavailability and microbial activity, which leads to enhanced microbial degradation of pyrene.     

Variations in competitive mechanisms of captive male hamadryas-like baboons in two feeding situations

Experiments were carried out to determine the influence of food distribution (clumped vs dispersed) on processes of competition among seven captive male hamadryas-like baboons. Spatial cohesion, cohesive behaviours that males performed and received from their females, and aggressive and feeding behaviours of the males were collected during 35 min after food supply. Median values and variability of these variables were compared in both the feeding situations. Food distribution influenced both asymmetries in access and the form of competition. When food was clumped, those with less access fed by means of compensatory or alternative strategies (away from the main source and/or by feeding supplants), and they were more aggressive towards other males. When food was dispersed, all individuals could feed simultaneously, males that were more aggressive were those with less cohesive OMUs, despite the fact that cohesive behaviours remained constant. Females appeared to contribute to variation in the OMU's cohesion through their responses to both food distribution and males' competitive ability.     

Theobromine intoxication in a red fox and a European badger in Sweden

A red fox (Vulpes vulpes) and a European badger (Meles meles) were found dead on a golf-course in October 1997 near Stockholm (Sweden). At necropsy, both animals were obese and the main finding was acute circulatory collapse. Theobromine intoxication was suspected as chocolate waste was available at a nearby farm and no other cause of death could be detected. Gastric contents and samples of liver from both animals were analyzed by reversed-phase high pressure liquid chromatography for the presence of methylxanthines. Theobromine and caffeine were detected in gastric contents and theobromine was identified in the liver samples from both animals. This appears to be the first report of theobromine intoxication in the red fox and the European badger.     

Muscarinic Cholinergic Receptor Subtypes in Hippocampus in Human Cognitive Disorders

Total muscarinic receptor levels, the levels of the subtypes exhibiting high and low affinity for pirenzepine, and the high- and low-affinity agonist states of the receptor were investigated in hippocampal tissue obtained at autopsy from mentally normal individuals and the following pathological groups: Alzheimer's disease, Parkinson's disease, Down's syndrome, alcoholic dementia, Huntington's chorea, and motor-neurone disease. A moderate decrease in the density of both high-affinity pirenzepine and high-affinity agonist subtypes was found in Alzheimer's disease, whereas a trend towards an increase in the overall muscarinic receptor density was apparent in the parkinsonian patients without dementia, mainly due to an increase in the low-affinity agonist state; the differences between the Alzheimer's disease and nondemented parkinsonian cases were highly significant. As previously reported, the levels of both choline acetyltransferase and acetylcholinesterase were markedly reduced in both Alzheimer's disease and Parkinson's disease--with a greater loss of both enzymes in the demented subgroup of parkinsonian patients. Activities of the cholinergic enzymes were also extensively reduced in Down's syndrome, accompanied by a loss of high-affinity pirenzepine binding. There were no significant receptor or enzyme alterations in the other groups studied. These observations suggest that in the human brain, extensive degeneration of cholinergic axons to the hippocampus, as indicated by a loss of cholinergic enzymes, is not necessarily accompanied by extensive muscarinic receptor abnormalities (as might be expected if a major subpopulation were presynaptic). Moreover, the opposite changes in muscarinic binding in Parkinson's and Alzheimer's diseases may be related to the greater severity of dementia in the latter disease.     

Landscape configuration and diversity hotspots in wintering sparrows

Community-level studies with finches have traditionally viewed local resources as the primary constrainst on local diversity. Patches have been considered to be self-contained and embedded in landscapes that were neutral with respect to the ecological processes under investigation. This study uses a factorial design to examine the relative roles of patch content and patch context in determining patterns of species richness. Sparrows were surveyed in small fallow patches that varied in both weed cover and the type of adjacent habitat. Species richness and total sparrow abundance were significantly influenced by both factors. Individual species were also influnced by both factors; however, responses were species-specific. Because occupation of particular plot types was conditional on their association with specific habitat types, the spatial patterning of species assemblages results from the configuration of patch types within the landscape.     

Embryo-uterine interaction in implantation

Pregnant donor (day 3) and non-pregnant recipient rats were hypophysectomized and injected daily for 6 days with 2 mg of progesterone. A single dose of 20 ng of estradiol-17β in saline was administered via a tail vein to either the donor, the recipient, or to both animals; blastocysts were transferred 60 to 90 minutes after the latter injection. Twenty-four hours later uterine implantation sites were delineated by injection of Chicago Blue-B dye. The results indicate that both the blastocyst and the uterus must be exposed to estrogen to obtain normal implantation rates. While 43.2% of the embryos implanted when both the donor and the recipient received estrogen, only 6.3% implanted when only the recipient was injected with estrogen. No implantations were found in animals in which only the embryos had been exposed to estrogen, suggesting that if this steroid was synthesized by the embryo it was insufficient to induce implantation in the rat.     

Identification of heme-binding proteins in the cell membranes of Vibrio anguillarum

Abstract Two strains of Vibrio anguillarum belonging to O1 and O2 serotypes were examined for their ability to bind hemin and hemoglobin. Whole cells as well as membrane extracts from both strains could clearly bind hemin and hemoglobin constitutively. Hemoglobin binding was completely inhibited by a 100-fold excess of unlabelled hemoglobin and also by hemin, suggesting the existence of specific receptors for heme groups in the cell membranes. Several hemin-binding and hemoglobin-binding bands with similar molecular sizes were detected in polyacrylamide gels as well as in Western blots. Only two of these protein bands in both strains were iron-regulated while the others were independent of the cell iron status. We conclude that both serotypes of V. anguillarum possess heme-binding abilities by means of membrane proteins.     

Cleavage of mitogen-activated protein kinases in human neutrophils undergoing apoptosis: role in decreased responsiveness to inflammatory cytokines

Extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) are major signaling molecules activated in human neutrophils stimulated by cytokines. Both molecules were cleaved at the N-terminal portion in neutrophils undergoing apoptosis induced by in vitro culture alone or treatment with TNF and/or cycloheximide. The cleavage of both molecules was inhibited by G-CSF and benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone, a caspase inhibitor, both of which can inhibit neutrophil apoptosis. In a cell-free system, ERK and p38 MAPK were not cleaved by recombinant caspase-3 or caspase-8 while gelsolin was cleaved by caspase-3 under the same condition. The cleavage of both molecules appears to be specific to mature neutrophils, since it was not detected in immature cells (HL-60 and Jurkat) undergoing apoptosis, indicating that proteases responsible for the cleavage of both molecules may develop during differentiation into mature neutrophils. Concomitant with the cleavage of ERK and p38 MAPK, GM-CSF- and TNF-induced superoxide release, adherence, and phosphorylation of ERK and p38 MAPK were decreased in neutrophils undergoing apoptosis. In addition, GM-CSF- and TNF-induced superoxide release and adherence were inhibited by PD98059 MAPK/ERK kinase inhibitor) as well as SB203580 (p38 MAPK inhibitor), suggesting possible involvement of ERK and p38 MAPK in superoxide release and adherence induced by these cytokines. These findings indicate that ERK and p38 MAPK are cleaved and degraded in neutrophils undergoing apoptosis in a caspase-dependent manner and the cleavage of both molecules may be partly responsible for decreased functional responsiveness to inflammatory cytokines.     

Effects of iso-osmotic NaCl and mannitol on growth, proline content, and antioxidant defense in Mammillaria gracilis Pfeiff. in vitro-grown cultures

Effects of iso-osmotic concentrations of NaCl and mannitol were studied in Mammilaria gracilis (Cactaceae) in both calli and tumors grown in vitro. In both tissues, relative growth rates were reduced under osmotic stress, which were accompanied by a decrease in both tissue water and K⁺ content. However, growth was inhibited to a lesser extent after exposure to NaCl, when accumulation of Na⁺ ions was observed. In calli, only salinity increased proline content, whereas with tumors proline accumulated after both osmotic stresses. Osmotic stresses also induced oxidative damage in both cactus tissues, although higher oxidative injury was caused by mannitol in calli and by salt in tumors. Low iso-osmotic concentrations of NaCl (75 mM) and mannitol (150 mM) increased peroxidase, ascorbate peroxidase, and esterase activities, whereas elevated catalase activity was recorded only after mannitol treatment in both tissues. High osmotic stress generally decreased enzymatic activities. However, in calli, esterase activity increased in response to high salinity, whereas ascorbate peroxidase activity was enhanced after high mannitol stress. In conclusion, both in vitro-grown cactus tissues were found to be sensitive to osmotic stress caused by either mannitol or NaCl, but accumulation of Na⁺ ions in response to salt somewhat contributed to osmotic adjustment. However, more prominent oxidative damage induced by NaCl compared to mannitol in tumor could be related to ion toxicity. The mechanisms that mediate responses to salt- and mannitol-induced osmotic stresses differed and were dependent on tissue type.     

Altered insulin signaling in retinal tissue in diabetic states

Both type 1 and type 2 diabetes can lead to altered retinal microvascular function and diabetic retinopathy. Insulin signaling may also play a role in this process, and mice lacking insulin receptors in endothelial cells are protected from retinal neovascularization. To define the role of diabetes in retinal function, we compared insulin signaling in the retinal vasculature of mouse models of type 1 (streptozotocin) and type 2 diabetes (ob/ob). In streptozotocin mice, in both retina and liver, insulin receptor (IR) and insulin receptor substrate (IRS)-2 protein and tyrosine phosphorylation were increased by insulin, while IRS-1 protein and its phosphorylation were maintained. By contrast, in ob/ob mice, there was marked down-regulation of IR, IRS-1, and IRS-2 protein and phosphorylation in liver; these were maintained or increased in retina. In both mice, Phosphatidylinositol 3,4,5-trisphosphate generation by acute insulin stimulation was enhanced in retinal endothelial cells. On the other hand, protein levels and phosphorylation of PDK1 and Akt were decreased in retina of both mice. Interestingly, phosphorylation of p38 mitogen-activated protein kinase and ERK1 were responsive to insulin in retina of both mice but were unresponsive in liver. HIF-1alpha and vascular endothelial growth factor were increased and endothelial nitric-oxide synthase was decreased in retina. These observations indicate that, in both insulin-resistant and insulin-deficient diabetic states, there are alterations in insulin signaling, such as impaired PDK/Akt responses and enhanced mitogen-activated protein kinases responses that could contribute to the retinopathy. Furthermore, insulin signaling in retinal endothelial cells is differentially altered in diabetes and is also differentially regulated from insulin signaling in classical target tissues such as liver.     

Sex differences in social stress-induced pressor and behavioral responses in normotensive and prehypertensive rats

This study investigated sex differences in chronic social stress-induced pressor and behavioral responses in normotensive and prehypertensive rats. Adult Wistar and borderline hypertensive (BH) rats (offspring of Wistar dams and spontaneously hypertensive sires) of both sexes were exposed to crowding stress (200 cm2/rat, 5 rats/cage) for 6 weeks. Controls were kept 4 rats/cage (480 cm2/rat). Blood pressure (BP) and open field activity were determined before experiment and after 1, 3 and 6 weeks of stress. Basal BP of BH rats was higher than in Wistar (p < 0.001) in both males and females. Horizontal and vertical activity of BH males and females was elevated vs. Wistar (p < 0.01) and females in both phenotypes were more active than the respective males (p < 0.01). Crowding resulted in delayed between-session habituation and significant elevation of BP only in BH males (143 ± 2 vs. 134 ± 3 mmHg in controls after 6-week crowding). No changes of BP were observed in crowded females of both phenotypes regardless of their delayed between-session habituation. Thus chronic social stress produced by crowding seems to represent a significant risk factor for development of stress-related hypertension only in males with genetic predisposition to high blood pressure while females of both phenotypes responded to stress by impaired between-session habituation.     

Chromosomal changes in two strains of human fibroblasts cultured in vitro

The chromosomal changes that occurred in two freshly initiated human cell strains, during their in vitro life were followed. Both strains are characterized by slow growth, and both show a period of delayed growth. In one of the strains the chromosomal composition showed changes just before the period of slow growth. After this period both cell strains showed several kinds of aneuploid chromosome complements, mainly in the hypotetraploid region. Aberrant chromosomes were present at the end of the in vitro life of both strains.The work in this paper was supported by the Foundation for Basic Medical Research (FUNGO), and the Association between Euratom and the University of Leiden, contract No. 052-64-1-BIAN.