Immune Control of Chlamydial Growth in the Human Epithelial Cell Line RT4 Involves Multiple Mechanisms That Include Nitric Oxide Induction,Tryptophan Catabolism and Iron Deprivation

The antimicrobial activity of T cell-derived cytokines, especially interferon (IFN)-γ, against intracellular pathogens, such as Chlamydia trachomatis, involves the induction of 3 major biochemical processes: tryptophan catabolism, nitric oxide (NO) induction and intracellular iron (Fe) deprivation. Since the epithelial cell is the natural target of chlamydial infection, the presence of these antimicrobial systems in the cell would suggest that they may be involved in T cell control of intracellular multiplication of Chlamydia. However, the controversy over whether these 3 antimicrobial processes are present in both mice and humans has precluded the assessment of the relative contribution of each of the 3 mechanisms to chlamydial inhibition in the same epithelial cell from either mice or humans. In the present study, we identified a Chlamydia-susceptible human epithelial cell line, RT4, that possesses the 3 antimicrobial systems, and we examined the role of nitric oxide (NO) induction, and deprivation of tryptophan or Fe in cytokine-induced inhibition of chlamydiae. It was found that the 3 antimicrobial systems contributed to cytokine-mediated inhibition of the intracellular growth of Chlamydia. NO induction accounted for ~20% of the growth inhibition; tryptophan catabolism contributed approximately 30%; iron deprivation was least effective; but the combination of the 3 systems accounted for greater than 60% of the inhibition observed. These results indicate that immune control of chlamydial growth in human epithelial cells may involve multiple mechanisms that include NO induction, tryptophan catabolism and Fe deprivation.     

Effects of temperature and ultraviolet radiation on microbial foodweb structure: potential responses to global change

1. A series of growth experiments were conducted with natural plankton communities from a lake and river in northern Quebec, to evaluate the response of microbial foodweb structure to changes in ambient temperature and solar ultraviolet radiation (UVR). 2. Bioassays were incubated for 6 days at two temperatures (10 and 20 °C) and three near-surface irradiance conditions [photosynthetically active radiation (PAR) + UVA + UVB, PAR + UVA, and PAR only). 3. The concentration of total bacteria showed no net response to temperature, but the percentage of actively respiring bacteria, as measured with the cellular redox probe 5-cyano-2,3-ditolyl tetrazolium chloride (CTC), was up to 57% higher at 20 °C relative to 10 °C. Chlorophyll a concentrations in the < 2 μm size fraction also reacted strongly to temperature, with a net increase of up to 61% over the temperature range of 10–20 °C. 4. The UVR effects were small or undetectable for most of the measured variables; however, the percentage of actively respiring bacteria was significantly inhibited in the presence of UVR at 20 °C, decreasing by 29–48% on day 6 in the lake experiments and by 59% on day 2 in one river experiment. 5. The results show differential sensitivity to temperature among organisms of microbial communities in subarctic freshwaters, and a resilience by the majority of micro-organisms to their present UVR conditions. Microbial foodwebs in northern freshwaters appear to be relatively unresponsive to short-term (days) changes in UVR. However, the observed responses to temperature suggest that climate change could influence community structure, with warmer temperatures favouring picoplanktonic phototrophs and heterotrophs, and a shift in nanoplankton species composition and size structure.     

Effects of fat adaptation and carbohydrate restoration on prolonged endurance exercise.

We determined the effect of fat adaptation on metabolism and performance during 5 h of cycling in seven competitive athletes who consumed a standard carbohydrate (CHO) diet for 1 day and then either a high-CHO diet (11 g. kg(-1)x day(-1) CHO, 1 g x kg(-1) x day(-1) fat; HCHO) or an isoenergetic high-fat diet (2.6 g x kg(-1) x day(-1) CHO, 4.6 g x kg(-1) x day(-1) fat; fat-adapt) for 6 days. On day 8, subjects consumed a high-CHO diet and rested. On day 9, subjects consumed a preexercise meal and then cycled for 4 h at 65% peak O(2) uptake, followed by a 1-h time trial (TT). Compared with baseline, 6 days of fat-adapt reduced respiratory exchange ratio (RER) with cycling at 65% peak O(2) uptake [0.78 +/- 0.01 (SE) vs. 0.85 +/- 0.02; P < 0.05]. However, RER was restored by 1 day of high-CHO diet, preexercise meal, and CHO ingestion (0.88 +/- 0.01; P < 0.05). RER was higher after HCHO than fat-adapt (0.85 +/- 0.01, 0.89 +/- 0.01, and 0.93 +/- 0.01 for days 2, 8, and 9, respectively; P < 0.05). Fat oxidation during the 4-h ride was greater (171 +/- 32 vs. 119 +/- 38 g; P < 0.05) and CHO oxidation lower (597 +/- 41 vs. 719 +/- 46 g; P < 0.05) after fat-adapt. Power output was 11% higher during the TT after fat-adapt than after HCHO (312 +/- 15 vs. 279 +/- 20 W; P = 0.11). In conclusion, compared with a high-CHO diet, fat oxidation during exercise increased after fat-adapt and remained elevated above baseline even after 1 day of a high-CHO diet and increased CHO availability. However, this study failed to detect a significant benefit of fat adaptation to performance of a 1-h TT undertaken after 4 h of cycling.     

Burrowing owls and burrowing mammals: are ecosystem engineers interchangeable as facilitators?

Terrestrial vertebrates exhibit dynamic, positive interactions that form and dissolve under different circumstances, usually with multiple species as participants. Ecosystem engineers are important facilitators of other species because they cause physical changes in the environment that alter resource availability. Although a species can be associated with more than one partner, facilitators may not be interchangeable if they differ in abundance, behavioral characteristics, or interactions with other factors in ways that condition the outcome of the association. We examined interactions between burrowing owls (Athene cunicularia) and two burrowing mammals, hairy armadillos (Chaetophractus villosus) and plains vizcachas (Lagostomus maximus), and determined whether these ecosystem engineers are interchangeable for owls. We examined reproductive success for owls nesting in these mammal burrows, constructed a logistic regression model to identify habitat characteristics associated with owl nests, and examined the engineering activities of the mammals. Data on reproduction and habitat indicate that armadillos and vizcachas are not interchangeable for owls. Thirty-five percent of the nests in vizcacha burrows produced fledglings; no fledglings were produced from nests outside vizcachas colonies, even though owls nest successfully in armadillo burrows in other parts of Argentina. Vizcachas facilitate burrowing owls by construction of burrows and by producing open understory vegetation through herbivory. In contrast, armadillos do not alter vegetation, and their burrows are suitable for nest sites only when they occur in recently burned areas or areas maintained by anthropogenic disturbance. Our habitat model also suggests that fire plays a key role in maintaining owl populations because fire is the only natural process that reduces shrubs to the level required by owls. Current management practices of eradication of vizcachas and fire suppression in shrublands could have strong negative consequences for burrowing owls.     

HYBRID FITNESS IN THE LOUISIANA IRISES: ANALYSIS OF PARENTAL AND F1 PERFORMANCE

The assumption of hybrid inferiority is central to the two models most widely applied to the prediction of hybrid zone evolution. Both the tension zone and mosaic models assume that natural selection acts against hybrids regardless of the environment in which they occur. To test this assumption, we investigated components of fitness in Iris fulva, I. hexagona and their reciprocal F₁ hybrids under greenhouse conditions. The four cross types were compared on the basis of seed germination, vegetative and clonal growth, and sexual reproduction. In all cases, the hybrids performed as well as, or significantly better than, both of their parents. These results suggest that F₁ hybrids between I. fulva and I. hexagona are at least as fit as their parents. The results of this study are therefore inconsistent with the assumptions of both the tension zone and mosaic models of hybrid zone evolution.     

X-Linked chronic granulomatous disease: mutations in the CYBB gene encoding the gp91-phox component of respiratory-burst oxidase.

Chronic granulomatous disease (CGD) is a hereditary disorder of host defense due to absent or decreased activity of phagocyte NADPH oxidase. The X-linked form of the disease derives from defects in the CYBB gene, which encodes the 91-kD glycoprotein component (termed "gp91-phox") of the oxidase. We have identified the mutations in the CYBB gene responsible for X-linked CGD in 131 consecutive independent kindreds. Screening by SSCP analysis identified mutations in 124 of the kindreds, and sequencing of all exons and intron boundary regions revealed the other seven mutations. We detected 103 different specific mutations; no single mutation appeared in more than seven independent kindreds. The types of mutations included large and small deletions (11%), frameshifts (24%), nonsense mutations (23%), missense mutations (23%), splice-region mutations (17%), and regulatory-region mutations (2%). The distribution of mutations within the CYBB gene exhibited great heterogeneity, with no apparent mutational hot spots. Evaluation of 87 available mothers revealed X-linked carrier status in all but 10. The heterogeneity of mutations and the lack of any predominant genotype indicate that the disease represents many different mutational events, without a founder effect, as is expected for a disorder with a previously lethal phenotype.     

Do ecto- and ericoid mycorrhizal fungi produce peroxidase activity?

 Several reports attest to the apparent ability of some ectomycorrhizal (ECM) and ericoid (ERM) mycorrhizal fungi to produce peroxidase enzyme activities during growth in axenic culture. In critically reviewing these data, we highlight that apparent peroxidase activities have been observed during growth in media containing 60–70 μM Fe. ECM and ERM fungi are known to produce H₂O₂ via carbohydrate oxidase activity and conditions in common culture media are favourable to the production of hydroxyl radicals, superoxide radicals and ferryl ions via the Fenton reaction. Free radicals so produced can mediate oxidation of substrates commonly used in presumptive peroxidase assays, leading to false-positive results. We argue that there is currently no evidence to support production of peroxidase activity by ECM or ERM fungi, but highlight circumstances in which peroxidase expression might be observed in future work. Accepted: 16 July 1998     

Trophic roles of particle feeders and detritus in a mangrove island prop root ecosystem

Swarms of Dioithona oculata, shoals of Mysidium columbiae, and schools of planktivorous fish were observed among mangrove prop roots on a lagoonal island. Detritus covering epiphytic and benthic macroalgae and invertebrates was a probable food source for pelagic mysids, and copepods, as well as benthic oysters and sponges. Open water blooms of dinoflagellate Amphidinium klebsii were probably only accessible to the swarming dioithonans which spend the night away from the mangrove prop roots. Dominant species were analyzed for stable carbon isotopic composition (expressed as ¹³C%.) to elucidate the origin and fate of detritus. Detrital components (–23.6 to –19.1%.), which included floating detritus, marine snow, prop root detritus, and detritus below prop roots, probably originated from the dominant subtidal macroalgal species (–24.2 to –14.6%.) and A. klebsii (–25.8 to –21.2%.), although other sources which are end-members could theoretically contribute since detrital isotopic ratios are half way between end-members. These other sources include the intertidal red algal Bostrychia spp. (–30.9 to –29.6%.), red mangrove Rhizophora mangle leaves (–28.2 to –27.0%.), and turtle grass Thalassia testudinum (–12.3 to –11.1%.). Particle feeders such as mangrove oysters, sponges, mysids, and dioithonans were usually enriched with ¹³C (–21.0 to –16.2%.) compared to their probable food sources, detritus and A. klebsii. At Anchovy Bay, adult mysids and planktivorous fish were more ¹³C enriched (1–3%.) than their probable prey, the dioithonans. Turtle grass may have a minor role in food webs, since dioithonans and mysids from Outer Twin and Anchovy Bays where turtle grass beds were abundant were consistently more ¹³C enriched (2–7%.) than in the Lair Channel where turtle grass beds were sparse.