Observations on lesser-known flatworms: temnocephala

Temnocephala novae-zealandie, a flatworm epizoic on crayfish, was examined by light and electron microscopy to investigate the variation within rhabdocoel turbellarians and to provide information on the possible structural modifications in the evolution of parasitism. The sucker is clearly glandular; the tentacular glands are eosinophilic and at the surface store and often release mucus as a coiled thread; the epidermis is clearly not reduced in structure and contains septate junctions in the anterior portion. Light microscope studies documented the presence of ten pair of paranephrocytes (athrocytes). In the laboratory, egg deposition, survival on and apart from the host and another crayfish, and behavior during the host molt were observed.     

Effects of capture on biological parameters in free-ranging bighorn sheep (Ovis canadensis): evaluation of normal, stressed and mortality outcomes and documentation of postcapture survival

Blood samples and physiological data were collected from 634 bighorn sheep (Ovis canadensis) captured by four different methods between 1980 and 1986 in the western United States. These parameters were evaluated for selected physiological, biochemical and hematological values. Postcapture biological parameters were compared among bighorn sheep according to four different outcomes; normal, stressed or compromised, capture myopathy (CM) mortality, and accidental mortality. Significant differences (P less than 0.05) were noted between outcome groups relative to certain parameters: temperature, respiration, creatinine phosphokinase (CPK), lactic dehydrogenase (LDH), serum glutamic oxaloacetic transaminase (SGOT), blood urea nitrogen (BUN), glucose, white blood cell count (WBC) and plasma pH. Such differences between groups may help in evaluating the clinical status of bighorn sheep at capture, enabling one to predict those animals that might develop CM at a later date, indicate candidates for preventive medical treatment prior to release, and/or which should be followed closely to determine long-term survival. Evaluation of follow-up data (n = 77) related to outcome status and long-term survival of bighorn sheep indicated that less than 4% (3 of 77) were dead within 1 mo of capture (one of these had been classified as normal and two as stressed or compromised at capture); less than 3% (3 of 77) were dead greater than 1 mo, and less than 6 mo after capture two were classified in the stressed outcome and one as diseased. Eighty-eight percent (68 of 77) were alive from 1 mo to 5 yr after capture (53 were classified as normal, 12 as stressed or compromised and 3 as diseased), and 2% (1 of 77) had chronic CM but was still alive (this animal had been classified as normal). Of 77 sheep in the follow-up group, less than 3% (2 of 77) were not observed following capture (one was classified as normal and one as stressed and diseased). Of the fatalities, less than 3% (2 of 40) had been captured by the net-gun and less than 4% (1 of 27) by drive-net. Those two unobserved in the follow-up group also had been caught with the net-gun, 5% (2 of 40). The single surviving CM case had been captured by the net-gun. Although the net-gun appears to be one of the safest methods of capturing individual bighorn sheep, based on evaluation of capture data and biological parameters, it may not be associated with the best long-term survival in some bighorn sheep.(ABSTRACT TRUNCATED AT 400 WORDS)     

Seasonal occurrence of Campylobacter spp. in surface waters and their correlation with standard indicator bacteria

Campylobacter jejuni, Campylobacter coli, and a Campylobacter-like organism were isolated from a number of natural water sources in central Washington, including ponds, lakes, and small mountain streams at elevations ranging from 1,460 to 5,400 feet (ca. 445 to 1,646 m) above sea level. At the two sites where extensive sampling was done, the bacteria were recovered throughout the year. Generally, the recovery rates were highest in the fall and winter months and lowest during the spring and summer months. Campylobacter density did not show significant correlation with microbiological (plate counts of fecal and total coliforms, fecal streptococci, and heterotrophic bacteria) or physical (water temperature, pH, and conductivity) parameters.     

Cell-mediated immunity in protection and pathology of malaria

The stimulation of protective immunity against malaria is the goal of many research groups. But trials with antigens that stimulate antibodies have yet to fulfil these expectations, and it is increasingly recognized that non-antibody-mediated immunity is also important in immunity to malaria - especially through mediators such as gamma interferon, tumour necrosis factor and reactive forms of oxygen. However, the host can suffer if this type of immune response is too exuberant, and in this review, Ian Clark argues that much of what is recognized as clinical malaria is caused in this way. He suggests that only when discussed in these terms can malaria illness and pathology be seen as a coherent, predictable entity instead of a sea of unconnected surprises. Moreover, these ideas have important implications for vaccine development that, although requiring more basic work, must not be neglected.     

Triphenylethylene antiestrogen binding sites (TABS) specificity

The relative binding affinities (RBA) of various compounds for the triphenylethylene antiestrogen binding sites (TABS) were examined. The ability of tamoxifen to inhibit the binding of [3H]tamoxifen to salt extracted (0.4 M KCl) TABS from rat liver nuclei was used as a standard by which other compounds were compared (tamoxifen RBA, 100; Kd approximately 1 nM). Nafoxidine was the most effective triphenylethylene compound used (RBA 333; Kd approximately 0.3 nM) whereas the RBA of zuclomiphene and enclomiphene was not different from tamoxifen. MER-29 was the weakest inhibitor of the triphenylethylene derivatives (RBA 10; Kd approximately 10 nM). Trifluoperazine, chlorpromazine and the anti-calmodulin drugs W-13 and W-12 had RBA's of 25, 1, 1 and 0.1 respectively. The binding affinities of cholesterol and 7-ketocholesterol were significant (Kd approximately 22 nM) while the steroid hormones, estradiol, testosterone, progesterone and corticosterone displayed not observable affinity. Various compounds obtained from Merrill Dow Pharmaceuticals and the Eli Lilly Company which contained alklaminoethoxy side chains linked to aromatic ring structures had RBA's ranging from 1-0.3. We conclude, as other investigators have also concluded, that the similar binding affinities of various triphenylethylene antiestrogens for TABS and their divergent activities as antiestrogens makes it unlikely that TABS are directly involved in estrogen antagonism. The moderate but significant affinity of TABS for trifluoperazine and other drugs thought to be involved in calmodulin regulation indicates that TABS may be a linked in some way to calmodulin function. The binding of cholesterol and 7-ketocholesterol is also significant and may indicate that TABS are involved in cholesterol metabolism.     

The effects of ionizing radiation on the pulmonary vasculature of intact rats and isolated pulmonary endothelium

We studied the effects of ionizing radiation on the morphology of the pulmonary circulation using an in vivo rat model and an in vitro pulmonary artery endothelial cell model. Gamma radiation was given as either an acute (30 Gy) or fractionated (5 X 6 Gy) dose to one hemithorax of rats. An acute 30-Gy dose delivered resulted in a 70% decrease in pulmonary arterial perfusion, using technetium-99m microaggregated albumin (99mTc-MAA), in the irradiated lung by 2-3 weeks after irradiation. Pulmonary microradiographs, using a barium sulfate perfusion method, obtained 2-3 weeks after irradiation demonstrated widespread loss of capillary filling and segmentation of the vessels. Histologic examination demonstrated intact capillaries, suggesting that the alterations in pulmonary perfusion were at the precapillary level. Similar abnormalities in lung perfusion and morphology were found after delivery of fractionated doses of radiation, but the onset of the changes was delayed, occurring 4-6 weeks postirradiation. Using cultured pulmonary endothelial cell monolayers, cell sloughing and retraction from the surface substrate were observed within 24 h after in vitro delivery of 30 Gy. Similar findings occurred in monolayers given fractionated doses (5 X 6 Gy) of radiation 2-3 days after the final dose. The in vivo animal and in vitro endothelial cell models offer a useful means of examining the morphologic alterations involved in radiation lung vascular damage.     

Identification of a novel adhesion molecule in human leukocytes by monoclonal antibody LB-2

Monoclonal antibody LB-2 to a surface antigen on human B cells, lymphoblast, monocytes and vascular endothelial cells largely inhibited adhesion among Epstein Barr virus-immortalized normal B cells (EBV-B) and concanavalin A-stimulated blood mononuclear cells (Con A-BMC) before and after phorbol ester treatment. The antibody inhibited to a lesser extent phorbol ester-induced aggregation of monocytes, U937 cells and fresh BMC and had virtually no inhibitory effect on the adhesion among enriched T cells and granulocytes. A surface glycoprotein band of 84 kDa was obtained from EBV-B cells by immunoprecipitation and gel electrophoresis. Immunological and biochemical studies clearly distinguished this molecule from gp90 and associated glycoproteins which also mediate leukocyte adhesion.     

A Pseudomonas stutzeri outer membrane protein inserts copper into N2O reductase

Among a set of frameshift mutagen (ICR-191; Polysciences, Inc.)-induced mutations that confer inability to grow anaerobically with N2O as the sole electron acceptor, one class was found that produced an inactive N2O reductase which lacked copper. All of these mutant strains failed to produce a 61,000-Mr protein located in the outer membrane. This protein, termed NosA, seems not to be responsible for bringing copper into the cell because the mutant strains and their parent were similarly sensitive to the copper content of the growth medium and no intermediate copper concentration in the medium permitted the mutant strains (nosA) to grow anaerobically with N2O as the sole electron acceptor. We conclude that NosA is necessary to insert copper into N2O reductase or to maintain it there.     

Stimulus-specific induction of phospholipid and arachidonic acid metabolism in human neutrophils

Phospholipid remodeling resulting in arachidonic acid (AA) release and metabolism in human neutrophils stimulated by calcium ionophore A23187 has been extensively studied, while data obtained using physiologically relevant stimuli is limited. Opsonized zymosan and immune complexes induced stimulus-specific alterations in lipid metabolism that were different from those induced by A23187. [3H]AA release correlated with activation of phospholipase A2 (PLA2) but not with cellular activation as indicated by superoxide generation. The latter correlated more with calcium-dependent phospholipase C (PLC) activation and elevation of cellular diacylglycerol (DAG) levels. When cells that had been allowed to incorporate [3H]AA were stimulated with A23187, large amounts of labeled AA was released, most of which was metabolized to 5-HETE and leukotriene B4. Stimulation with immune complexes also resulted in the release of [3H]AA but this released radiolabeled AA was not metabolized. In contrast, stimulation with opsonized zymosan induced no detectable release of [3H]AA. Analysis of [3H]AA-labeled lipids in resting cells indicated that the greatest amount of label was incorporated into the phosphatidylinositol (PI) pool, followed closely by phosphatidylcholine and phosphatidylserine, while little [3H]AA was detected in the phosphatidylethanolamine pool. During stimulation with A23187, a significant decrease in labeled PI occurred and labeled free fatty acid in the pellet increased. With immune complexes, only a small decrease was seen in labeled PI while the free fatty acid in the pellets was unchanged. In contrast, opsonized zymosan decreased labeled PI, and increased labeled DAG. Phospholipase activity in homogenates from human neutrophils was also assayed. A23187 and immune complexes, but not zymosan, significantly enhanced PLA2 activity in the cell homogenates. On the other hand, PLC activity was enhanced by zymosan and immune complexes. Stimulated increases in PLC activity correlated with enhanced superoxide generation induced by the stimulus.