Increased elongation of N-acetyllactosamine repeats in doubly glycosylated lysozyme with a particular spacing of the glycosylation sites

Lysozyme is an example of an extensively studied secretory enzyme. Glycosylated mutant human lysozyme has been used as a model in studies on the biosynthesis of N-acetyllactosamine repeats in N-linked oligosaccharides. We examined the biosynthesis of the repeats in two doubly glycosylated mutants and describe here a rapid purification and separation of singly and doubly glycosylated molecules. In one of the mutants, the elongation of the repeats is enhanced if the molecules are doubly glycosylated, but not if the carbohydrate is attached to either site individually. This enhancement is not seen in the other doubly glycosylated mutant. Since lysozyme is not structurally related to glycoproteins bearing carbohydrate with N-acetyllactosamine repeats, we propose that in multivalent substrates the synthesis of the repeats can be promoted by a proper spacing of the elongated carbohydrate antennae in addition to any role of the protein backbone.     

Combined fluorescence and photovoltage studies on chlorosome containing bacteria

Energy transfer kinetics, primary charge separation, antenna size and excitonic connectivity of photosynthetic units (PSU) in whole cells of Chloroflexus aurantiacus were studied at room temperature by ps-fluorescence and ps-photovoltage as well as by stationary fluorescence-spectroscopy and fluorescence induction measurements. The fluorescence decay kinetics measured at different wavelengths are in accordance with the currently accepted sequential energy transfer from the chlorosomes via the baseplates to the B808–866 complexes and the final trapping in the RC with time constants of 19 ± 2 ps, 40 ± 4 ps and 90 ± 9 ps, respectively. However, the quantitative analysis of fluorescence spectra and the occurrence of slow phases in the fluorescence decays reveal that in whole cells a significant fraction of BChl c in the chlorosome and of BChl a in the baseplate is unconnected. The photovoltage kinetics consisted of two electrogenic phases with time constants of 118 ± 5 ps and 326 ± 35 ps and comparable electrogenicities. The first phase is ascribed to trapping from the B808-866 complexes by P+H_A- formation and the second one to charge stabilization on a quinone acceptor. Fluorescence induction curves displayed a pronounced sigmoidicity, indicating efficient lateral energy transfer between neighbored PSUs and a dense packing of 19 reaction centers (RC) beneath one chlorosome. A quantitative analysis of the fluorescence-induction curves at different excitation wavelengths allows the estimation of pigment stoichiometries (i.e. antenna sizes): BChl c/RC 794 and B808/RC 15.     

Damp Rooms and Saying ‘Please’: Mimesis and Alterity in the Host Family Space in Study-Abroad Experiences

Study-abroad programs are a ‘globalizing project’ developed at the intersection of educational institutions seeking new income sources and the ideologies that the world is comprised of a mosaic of cultures and that ‘intercultural experience’ is valuable. While research on study-abroad programs often focuses on their effect on the students’ ‘global competence’ and language proficiency, this article focuses on the space, the space of host family in particular, produced in study-abroad programs. Study-abroad programs here are perceived in a wide sense to include various programs for students to go to another country to study.

Study-abroad guidebooks and researchers portray the host family's life as constituting a quintessential ‘native life’, in which study-abroad students should be immersed. However, based on ethnographic fieldwork in Aotearoa/New Zealand, this article argues that the distinction between ‘cultural difference’ and ‘cultural sameness’ is a construction, resulting in mutual accommodation and intolerant judgments, respectively. That is, when host parents recognised the ‘cultural difference’ of the foreign students’ behaviour, they often sought to understand meanings behind the behaviour, resulting in mutual accommodation. It turned the space into what Michael Taussig calls the ‘space between’, in which it is unclear who is imitating whom. When the host parents recognised ‘cultural sameness’ based on superficial similarity, however, their efforts to understand meanings behind behaviour was halted and, when the behaviour was considered problematic, led to the student being asked to move out. This article details an ironic space of study abroad where alterity inspired tolerance and similarity inspired intolerance.     

ddC- and 3TC-bis(SATE) monophosphate prodrugs overcome cellular resistance mechanisms to HIV-1 associated with cytidine kinase deficiency.

A 2',3'-dideoxycytidine (ddC)-resistant T-lymphoid cell line (MOLT-4/8rddC250), in which deoxycytidine kinase (dCK) gene-expression was decreased when compared with parental cells, has been selected. Cytotoxic and antiretroviral activity of ddC and 3TC was significantly lower in MOLT-4/8rddC250-than in parental MOLT-4/8 cells. ddC- and 3TC-bis(SATE)phosphotriesters completely overcame cellular resistance mechanisms and showed comparable both cytotoxic and antiretroviral activity in parental and ddC-resistant cells.     

Phenology of Lacanobia subjuncta (Lepidoptera: Noctuidae) in Washington and Oregon apple orchards

The phenology of Lacanobia subjuncta (Grote & Robinson) (Lepidoptera: Noctuidae) was investigated in 30 apple orchards in central Washington state and northeastern Oregon from 1998 to 2001 (57 total orchard-yr). Adult captures in pheromone-baited traps were fit to a Weibull distribution to model emergence of the first and second generations. Initial capture of first generation adults was observed at 216.2 +/- 2.6 degree-days (DD) (mean +/- SEM) from 1 March by using a base temperature of 6.7 degrees C. The model predicted that flight was 5 and 95% complete by 240 and 700 degree-days (DD), respectively. Monitoring of oviposition and hatch was used to establish a protandry plus preoviposition degree-day requirement of 160.0 +/- 7.7 DD, as well as to provide data to describe the entire hatch period. Egg hatch was 5 and 95% complete by 395 and 630 DD, respectively. The start of the second flight was observed at 1217.1 +/- 8.3 DD by using an upper threshold for development of 32 degrees C and a horizontal cutoff. The model indicated that the second flight was 5 and 95% complete by 1220 and 1690 DD, respectively. Second generation hatch was 5 and 95% complete by 1440 and 1740 DD, respectively. A discussion of the potential uses of these detailed phenology data in optimizing management strategies is presented.     

Decreased glycogen mobilization during ochratoxicosis in broiler chickens.

Graded doses of pure ochratoxin A (0, 0.5, 1.0, 2.0, 4.0, and 8.0 microgram of toxin per g of feed) were incorporated into a commercial diet which was fed to chickens from hatching to 3 weeks of age, at which time the experiments were terminated. Liver glycogen levels were elevated significantly (P less than 0.05) by 4.0 and 8.0 microgram/g but not lower doses. Glucagon stimulation of glycogen mobilization was inhibited at the same concentrations. Histopathological examination revealed cytoplasmic but not nuclear deposits of glycogen in cells at the periphery of liver lobes. These data demonstrated that ochratoxin inhibited glycogenolysis. Impaired ability to generate glucose from glycogen could account for the increased susceptibility to cold stress previously reported to occur in ochratoxicosis. Based on present and prior observations, it seems possible that ochratoxin induces a syndrome which mimics the glycogen storage disease of type X which is caused by a deficiency in the cyclic AMP-dependent enzyme of the glycogenolytic enzymatic cascade.     

The Cooperative Breeding System of the Red-cockaded Woodpecker

The breeding system of the red-cockaded woodpecker is described based on data collected over six years from a population of 500 marked individuals in the Sandhills of North Carolina. Male-female pairs were the most common social unit (59%), but 30% of social units contained one or more adult helpers, and 11% consisted of solitary males. Helpers were almost exclusively male: 27% of males remained in their natal group as helpers for at least one year, whereas only four (1%) females did. Most breeding females remained as breeders in the same group from one year to the next (56%), but a surprising number (12%) moved to another group. Many movements were related to incest avoidance or mate death, but 39% involved deserting a mate, usually following successful reproduction. We suggest that females sometimes are forced from groups by immigrants or other group members. The median distance of movements by adult females was only 1.3 km. In contrast to females, no breeding males switched groups. Survival of both breeding (76%) and helper (80%) males was higher than that of breeding females (69%). Males exhibited two distinct life-history strategies. Some remained as helpers on their natal territory for one or more years, and became breeders by inheriting breeding status in the natal group (17% per year) or by replacing a deceased breeder in a nearby group (13% per year, median distance moved 1.0 km). Other males dispersed from their natal group permanently during their first year. Some of these males were floaters at age one year, others were solitary, and a few became helpers in a non-natal group, but many were breeders. In contrast to males that first functioned as helpers, those that dispersed after fledging moved long distances (median dispersal distance 4.5 km), longer even than dispersing female fledglings moved (median distance 3.2 km). The habitat saturation model of the evolution of cooperative breeding is based on selection between the two life-history strategies exhibited by male red-cockaded woodpeckers. The model therefore may be tested directly with this species. Another indication that this model is appropriate for this species is the existence of a resource (cavity trees) that might provide an ecological basis for habitat saturation.     

Levamisole regulates the proliferation of murine liver T cells through Kupffer-cell-derived cytokines

 We have previously shown that levamisole increases the cytotoxic, cytostatic, and proliferative activity of murine nonparenchymal liver cells (NPC) in vitro. We have also shown that the nonadherent subpopulation of NPC, which are composed predominantly of T lymphocytes, is very responsive to this agent when administered to mice. Kupffer cells or immigrant macrophages are also responsive to levamisole but to a lesser extent. These findings prompted us to investigate changes in cytokine production by NPC following-treatment of mice with levamisole (25 mg/kg, i.p.), which may help explain the observed alterations in the immune functions of these cells. We found that levamisole treatment of mice causes a threefold increase in production of interferon (IFN) α/β by adherent NPC (more than 80% – 90% Kupffer cells) in vitro. When IFN α/β was added to cultured cells, it decreased the proliferative capacity of liver T cells in a dose-dependent manner. In contrast, the addition of anti-IFNα/β was shown to augment levamisole-induced proliferation of unfractionated NPC and Kupffer cells. NPC production of interleukin 1 (IL-1) and interleukin-6 (IL-6) in vitro was also increased threefold following treatment of mice with levamisole. IL-6 added in vitro to cells significantly augmented levamisole-induced proliferation of liver T cells while anti-IL-6 reduced proliferative activity to control levels. These findings suggested that IFNα/β, IL-6, and IL-1 play important regulatory roles in controlling the proliferative response of murine liver-associated T lymphocytes to levamisole. Finally, the proliferation of bone marrow cells was increased in mice given 5-fluorouracil (5FU). On the other hand, the proliferation of NPC was dramatically suppressed when 5FU was administered. However, the proliferation of these cells was restored when levamisole was given after 5FU. Received: 27 November 1995 / Accepted: 16 October 1996     

Maximizing the value of systematic reviews in ecology when data or resources are limited

Systematic reviews provide a rigorous, repeatable and quantitative method for assessing and synthesizing all available empirical evidence to evaluate a specific research, management, or policy question. They are particularly well suited for evaluating the effectiveness of environmental management actions, and thus for underpinning evidence‐based adaptive natural resource management. However, their current utility may be limited in countries like Australia, where both the amount of research relative to land area and of well‐monitored, active land management for environmental purposes are relatively low. Based on our experience conducting two of the first ecological systematic reviews in Australia, we have developed a number of recommendations for conducting systematic reviews in situations where resources and/or primary research data are limited. We discuss potential modification or augmentation of most aspects of the systematic review process including selection of a review team, question formulation, search strategy, data analysis, and the communication of results, as well as the inherent tradeoffs between systematic thoroughness and available resources that are involved in these changes. We hope that our recommendations will encourage more ecologists to undertake systematic reviews even if primary research and resources to conduct the review appear to be limited, as even a modified systematic review can provide more defensible evidence‐based guidelines for management of natural resources.