Nutrient responses to the liming of lake Gårdsjön

The acidified lakes Lake Gårdsjön and Lake Stora Hästevatten the reference lake have been monitored since 1979 and 1980 respectively. The lakes are situated in SW Sweden; in an area severly affected by acid deposition. Lake Gårdsjön was limed in spring 1982. This paper analyses changes in nutrient concentrations upon liming of Lake Gårdsjön. The liming of Lake Gårdsjön was followed by a slight increase in ammonium, nitrate, and dissolved organic nitrogen concentrations. A drastic decrease occurred in particulate nitrogen and particulate carbon, whereas dissolved organic carbon increased. Total phosphorus and particulate phosphorus concentrations were similar to pre-limed conditions. The long-term decrease in phosphorus concentration, exhibited by the reference lake, was not identified in Lake Gårdsjön after liming, but total phosphorus concentration was still less than half compared to Lake Gårdsjön in the early 1970's. Additional measures such as phosphorus fertilization, should in certain cases be considered in addition to liming if the goal is to restore lakes to their pre-acidic conditions.     

Heterotopic microvascular growth plate transplantation of the proximal fibula: an experimental canine model

The reconstructive potential of microvascular transplantation of skeletal growth plates was investigated through heterotopic transfers. The distal radius was resected in two series of puppies of a known large breed and substituted with a microsurgically revascularized transplant from the proximal fibula. Evaluation was conducted through serial roentgenograms, goniometric registration of joint mobility, volume measurements, histology, and fluorescent bone labeling. In the first series, development of neuropathic-like destruction of the weight-bearing graft ensued in the majority of the animals. In the second series, prolonged protection from weight bearing inhibited this destruction and resulted in hypertrophy of the revascularized epiphyseal end of the transplant but clearly reduced longitudinal growth, with only one transplant exhibiting longitudinal growth that exceeded 50 percent of the value for the control. This experiment demonstrates that skeletal growth plates possess a capacity for hypertrophy under the influence of increased loads. Whether this adaptability is sufficient to allow microvascular transplantation of growth plates to become a clinically useful procedure in children remains unclear. Further laboratory investigations are mandatory prior to clinical application of microvascular transfers of epiphyseal growth plates.     

Adoptive immunotherapy of murine cytomegalovirus adrenalitis in the immunocompromised host: CD4-helper-independent antiviral function of CD8-positive memory T lymphocytes derived from latently infected donors.

The ability of memory T lymphocytes derived from latently infected mice to control murine cytomegalovirus disease in the immunocompromised host was studied by adoptive transfer experiments. At a stage of pathogenesis when virus had already colonized target tissues, a therapeutic antiviral function could be ascribed to the CD8+ subset. This in vivo function was not restricted to sites in which intravenously infused lymphocytes usually are trapped or home in, such as the lungs or the spleen, respectively, but was also evident in the adrenal glands, a site to which antiviral effector cells have to specifically migrate. Specific infiltration of adrenal gland cortical tissue by donor-derived CD8+ memory T lymphocytes was demonstrated. CD4+ memory T lymphocytes had no antiviral effect by themselves and also were not required for the function of the CD8+ effector cells in this short-term immunotherapy model. These findings should help settle the debate about which subset of T lymphocytes comprises the effector cells that can directly control cytomegalovirus infection in the murine model system.     

Vascularized bone allografts: in vitro assessment of cell-mediated and humoral responses

The immunologic consequences of transplantation of vascularized bone allografts have not been previously characterized. In this study, knee allografts, both vascularized and nonvascularized, were transplanted from Lewis rats to Brown Norway rats across a strong histocompatibility barrier. A total of 66 transplants and 8 control animals were evaluated. The vascularized knee grafts consisted of 1 cm of proximal tibia and distal femur with a minimal muscular cuff isolated on the femoral vessels, and these were transplanted to a heterotopic, subcutaneous position on the abdominal wall of the recipient rat. Nonvascularized allografts (identical but without anastomoses) were transplanted for comparison. The cell-mediated response was measured by lymphocytotoxicity assay, and the humoral response was measured by cytotoxic antibody assay, both employing 51Cr-labeled target cells. The timing and intensity of the immune response differed according to the type of graft. The vascularized bone allografts generated significant cell-mediated and humoral responses as early as 5 days posttransplant. A significant humoral response in nonvascularized bone allografts was not apparent until day 14, while cell-mediated response in these grafts was variable. These findings were correlated with the histologic appearance of the grafted tissue. Cyclosporine, which was administered to one group of vascularized bone allografts, resulted in the suppression of both types of immune responses. The histologic appearance of this group resembled that of isografts transplanted as controls. The clinical application of vascularized bone allografts may offer significant advantages over nonvascularized allografts in the reconstruction of massive bone defects. Complications such as nonunion, fracture, and collapse of articular segments seen in nonvascularized allograft transplantation may be avoided by preservation of the blood supply to the graft. Characterization of the immune response to vascularized bone allografts may subsequently allow the manipulation of the host and/or graft tissue and promote graft incorporation.     

Evaluation of Diverse Antisera, Conjugates, and Support Media for Detecting Bradyrhizobium japonicum by Indirect Enzyme-Linked Immunosorbent Assay

We evaluated three antisera and four enzyme conjugates for the detection of Bradyrhizobium japonicum by an indirect enzyme-linked immunosorbent assay in microtiter plates. Nitrocellulose membrane sheets were then evaluated as an alternative support medium by using some combinations. Partially purified immunoglobulin G (IgG) or unpurified antisera to strain USDA 110 raised in rabbits, goats, or sheep was reacted in microtiter plates with alkaline phosphatase conjugated to protein A, goat anti-rabbit (GAR), sheep anti-rabbit (SAR), or rabbit anti-goat (RAG) IgG. Cultures or nodules containing homologous rhizobia were detected with equal sensitivity when protein A, GAR, or SAR was reacted with 5 μg of protein IgG per ml or a 1:800 titer of antisera from rabbits, but not goats or sheep. RAG reacted with IgG or antisera from goats or sheep. The detection limit was 2 × 10⁵ rhizobia per well. Rhizobia were spotted on nitrocellulose sheets as an alternative support medium, followed by soaking in 5 μg of protein per ml as IgG and 1:4,000 dilutions of protein A or GAR conjugate. Rhizobia in serogroup 110 were detected with the dye combination Nitro Blue Tetrazolium-5-bromo-4-chloro-3-indolyl phosphate (NBT-BCIP), and rhizobia in serogroup 122 were detected with fast red-naphthol phosphate (FR-NP). At the conclusion of the 5-h assay, purple (NBT-BCIP) or red (FR-NP) spots were visible in positive reactions. The sensitivity of detection was about 1,000 rhizobial cells or 3 μg of nodules tissue.     

Cell cycle traverse and protein metabolism in human NHIK 3025 cells: the role of anchorage

We have studied the effect of cell anchorage on the human cell line NHIK 3025 in vitro, to see whether the growth regulating effect of cell anchorage primarily affected DNA division cycle or mass growth cycle. It was found that cell to cell anchorage had the same effect on cell cycle progression as anchorage to a solid surface, which indicates that it is anchorage per se and not cell shape that is important for growth control in NHIK 3025 cells. When NHIK 3025 cells were grown without attachment to a solid surface, both G1 and cell cycle duration was prolonged by 6 h, which means that the prolonged cell cycle was due to a prolonged G1. During the first part of the cell cycle the rate of protein synthesis and degradation was constant, and at the same level in cells grown with and without attachment. This means that the prolonged G1 was not due to a reduced protein accumulation or mass growth. Towards the end of the cell cycle protein accumulation was reduced. This effect was either due to a size control before cell division or a secondary effect of the prolonged G1. We therefore conclude that cell anchorage as a growth regulator primarily affects the DNA/cell division cycle.     

Untersuchcungen zum Verhalten von Cryptococcus Neoformans in Erde

Zusammenfassung Innerhalb eines nachgewiesenermaßen mitCryptococcus neoformans (Cr. n.) infizierten Stadtgebietes wurden 170 Erdproben aufCr. n. untersucht. Während aus 6 von 50 Erdproben von Lokalisationen mit TaubenbesiedelungCr. n. nachgewiesen wurde, gelang dies bei keiner von 120 Proben ohne Taubenbesiedelung.Diese Engebnisse werden so gedeutet, daß zumindest unter den hiesigen BedingungenCr. n. als primärer Erdbodensaprophyt praktisch nicht vorkommt. Die Besiedelung von Taubenkot mitCr. n. geschieht vielmehr bereits im Darmkanalspontan infizierter Tauben, oder durch Kontakt mit infiziertem Kot. Hierbei gelangensekundär auch Kryptokokken in den Erdboden.Für künftige epidemiologische Untersuchungen wird eine Nachweismethodik empfohlen, die den Direktausstrich des in phys.-Kochsalzlösung suspendierten Untersuchungsmaterials auf Negersaat-Kreatinin-Antibiotika-Diphenylagar, seine Anreicherung in diphenylhaltiger (0,002 %) Bierwürze und den Tierversuch an der weißen Maus umfaßt. Mit dieser Methodik ist beim Vorhandensein von mindestens 10Cr. n. -Zellen in 1 g untersuchtem Ausgangsmaterial der Nachweis des Erregers zu erwarten.Halbquantitative Antrocknungsversuche vonCr. n. (ein stark und ein nahezu unbekapselter Stamm, zwei verschiedene Antrocknungsmethoden) an sterilen Seesand zeigten, daß selbst die unter direkter Einwirkung von Sonnenlicht angetrockneten Kryptokokken weitgehend überlebten.Die Direktkultur war dem Anreicherungsverfahren nahezu gleichwertig, während sich der Tierversuch im Vergleich dazu als weit weniger leistungsfähig erwies. Unter Hinweis auf frühere Untersuchungen wird betont, daß auf den Tierversuch dennoch nicht verzichtet werden sollte.

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Within the area of a city, well known to be infected withCryptococcus neoformans, 170 soil samples have been for this fungus investigated. While six out of 50 soil samples coming from pigeon locations were positive for the fungus, none of the other 120 soil samples became positive from areas without pigeon locations. For future epidemiologic investigations a special method is suggested. The direct culture of the material was almost equivalent with the method of enrichment, while the animal experiment in comparison was less efficient. It is very probable, thatCryptococcus neoformans is no primary germ of the soil. Soil becomes secondarily contaminated by pigeons and their excrements.

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Die Untersuchungen wurden dankenswerterweise durch Mittel des Bundesministers für Ernährung, Landwirtschaft und Forsten gefördert.     

Population regulation of a tropical damselfly in the larval stage by food limitation,cannibalism, intraguild predation and habitat drying

The relative importance of intraspecific, interspecific, and seasonal causes of larval mortality were investigated for aquatic larvae of the giant damselfly Megaloprepus coerulatus in Panama. These larvae live in water-filled holes in fallen and living trees, where they and three other common odonate species are the top predators. By mid wet season, M. coerulatus larvae were found in nearly half of all tree holes that harbored odonates. Although M. coerulatus were typically, but not always, eliminated from holes inhabited by larger hetero-specifics, M. coerulatus were more likely to encounter conspecifics than other odonate species. Hole with less than 11 of water rarely contained more than a single larva. In large holes where M. coerulatus was the only odonate species present, multiple larvae coexisted at a density of one larva per 1–21 of water. There the absence of 2–4 of the 5 larval size classes, despite a continuous input of eggs, suggested that cannibalism was a common cause of mortality. The size of the final instar, which determined adult body size, was correlated positively with tree hole volume for male, but not female, larvae. Experiments showed that when two larvae were placed together in 0.4–1 holes with abundant tadpole prey, the larger larva killed the smaller one. Often the larva that was killed was not eaten. Small larvae were more tolerant of each other than were pairs of medium or large larvae. Before killing occurred, the presence of larger larvae reduced the growth of smaller individuals, relative to controls. Obligate killing was density-dependent. In 3.0–1 holes with ad libitum prey, conspecific killing occurred until the larval density stabilized at one larva per 1–1.5 I, similar to the density found in large holes under field conditions, For M. coerulatus, cannibalism functions to reduce the number of potential competitors for food in addition to providing nutrition. When interactions between paired larvae in small holes were experimentally prevented, competition for food reduced the growth of one or both larvae relative to controls. Holes that were watered during the dry season supported larval densities similar to those in the wet season. Thus, dry season mortality could not be attributed to a decrease in available prey. Rather, M. coerulatus larvae could not survive more than 1 month of complete drying. Because the dry season typically lasts more than 6 weeks, habitat drying is a secondary source of mortality, affecting second- or third-generation larvae that fail to emerge before tree holes dry out completely.     

Nuclear and chloroplast transformation inChlamydomonas reinhardtii: strategies for genetic manipulation and gene expression

Transformation of the nuclear, chloroplast, and mitochondrial genomes can now be accomplished inChlamydomonas reinhardtii. Many biosynthetic pathways are carried out in the chloroplast, and efforts to manipulate these pathways will require that gene products be directed to this compartment. Chloroplast proteins are encoded in either the chloroplast or nuclear genome. In the latter case they are synthesized in the cytoplasm and imported post-translationally into the chloroplast. Thus, strategies for expressing foreign genes or overexpressing endogenous genes whose products reside in the chloroplast could involve either genome. This paper reviews the present status of transformation methodology for the nuclear and chloroplast genomes inChlamydomonas. Considerations for expressing gene products in the chloroplast are discussed. Experimental evidence for homologous recombination during transformation of the nuclear genome is presented.     

Morphological phylogenetics of theAsteraceae-Asteroideae,with notes on character evolution

The relationships within theAsteraceae-Asteroideae are scrutinized by means of cladistic analyses of morphological and chemical data. New data are presented, and novel interpretations of features previously used in cladistic analyses are made. Examples are insertion of anther filaments in the corolla, and morphology of disc floret styles, respectively. The results include a polyphyletic or paraphyleticInuleae s. l., and a paraphyleticHeliantheae s.l. in which theEupatorieae constantly form a subclade. Comparisons are made with previous cladistic analyses of morphological data, and those acquired from molecular data. Speculations about the evolution of certain organs are presented.