Methods and approaches to study plant mitochondrial alternative oxidase

The alternative oxidase is a non-proton motive 'alternative' to electron transport through the cytochrome pathway. Despite its wasteful nature in terms of energy conservation, the pathway is likely present throughout the plant kingdom and appears to be expressed in most plant tissues. A small alternative oxidase gene family exists, the members of which are differentially expressed in response to environmental, developmental and other cell signals. The alternative oxidase enzyme possesses tight biochemical regulatory properties that determine its ability to compete with the cytochrome pathway for electrons. Studies show that alternative oxidase can be a prominent component of total respiration in important crop species. All these characteristics suggest this pathway plays an important role in metabolism and/or other aspects of cell physiology. This brief review is an introduction to experimental methods and approaches applicable to different areas of alternative oxidase research. We hope it provides a framework for further investigation of this fascinating component of primary plant metabolism.     

Complementary and alternative therapies as treatment approaches for interstitial cystitis

The management of interstitial cystitis (IC) is predominantly the reduction of the symptoms of frequency, urgency, and pain. Multimodal treatment approaches for IC are helpful in customizing therapy for individual patients. Complementary and alternative therapies are a quintessential addition to the therapeutic armamentarium and frequently include dietary modification, nutraceuticals, bladder training, neuromodulation, stress reduction, and sex therapy. Dietary modification involves elimination of bladder irritants, fluid regulation, and a bowel regimen. Nutraceuticals studied for the treatment of IC include calcium glycerophosphate, L-arginine, mucopolysaccharides, bioflavinoids, and Chinese herbs. Bladder training is effective after pain reduction. The neuromodulation of high-tone pelvic-floor muscle dysfunction is achieved with physical therapy and acupuncture. Stress reduction and sex therapy are best administered by a qualified stress manager and sex therapist. Multimodal, nonconventional management may add efficacy to the treatment of IC.     

A comparison of marine protected areas and alternative approaches to coral-reef management

Marine protected areas (MPAs) have been widely adopted as the leading tool for coral-reef conservation, but resource users seldom accept them , and many have failed to produce tangible conservation benefits [3]. Few studies have objectively and simultaneously examined the types of MPAs that are most effective in conserving reef resources and the socioeconomic factors responsible for effective conservation [4-6]. We simultaneously explored measures of reef and socioeconomic conservation success at four national parks, four comanaged reserves, and three traditionally managed areas in Indonesia and Papua New Guinea. Underwater visual censuses of key ecological indicators [7, 8] revealed that the average size and biomass of fishes were higher in all areas under traditional management and at one comanaged reserve when compared to nearby unmanaged areas. Socioeconomic assessments [6, 9, 10] revealed that this effective conservation was positively related to compliance, visibility of the reserve, and length of time the management had been in place but negatively related to market integration, wealth, and village population size. We suggest that in cases where the resources for enforcement are lacking, management regimes that are designed to meet community goals can achieve greater compliance and subsequent conservation success than regimes designed primarily for biodiversity conservation.     

Peritonitis from Starch Glove Powder

Contamination of surgical wounds by starch glove powders has produced peritoneal inflammation and adhesions in many patients. Some have required surgical operation because of possible infection, others because of obstruction of the intestine. Resolving granulomas and fibrosis have been observed as incidental asymptomatic lesions in some patients during reoperation for other conditions.Starch granules are probably capable of producing inflammation in any patient, and the degree is believed to be related to the amount of starch present.Care in removing glove powder from gloves before they are worn in an operation will diminish or eliminate the problem. Removal can be accomplished by rubbing the gloves with a moist sterile towel or laparotomy tape.     

Mega-phylogeny approach for comparative biology: an alternative to supertree and supermatrix approaches

Background  

Biology has increasingly recognized the necessity to build and utilize larger phylogenies to address broad evolutionary questions. Large phylogenies have facilitated the discovery of differential rates of molecular evolution between trees and herbs. They have helped us understand the diversification patterns of mammals as well as the patterns of seed evolution. In addition to these broad evolutionary questions there is increasing awareness of the importance of large phylogenies for addressing conservation issues such as biodiversity hotspots and response to global change. Two major classes of methods have been employed to accomplish the large tree-building task: supertrees and supermatrices. Although these methods are continually being developed, they have yet to be made fully accessible to comparative biologists making extremely large trees rare.     

Combining palaeolimnological and limnological approaches in assessing lake ecosystem response to nutrient reduction

1. Palaeolimnological data and limnological time‐series data are highly complementary. Sediment records extend time‐scales, integrate subannual variability and expand the range of sites that can be studied, but they suffer from taphonomic biases and occasionally from uncertain chronology. Observational time‐series data, on the other hand, are highly resolved but are very limited in extent both in space and time. 2. Palaeolimnological and observational data‐sets need to be combined in oligotrophication research to establish (i) the past and present status of lakes needed to identify reference conditions; (ii) changes in ecosystem state; (iii) responses to nutrient reduction; and (iv) the potential role of other factors (e.g. additional stressors, climate change) that might confound predictions of future state.     

Cryopreservation of kangaroo spermatozoa using alternative approaches that reduce cytotoxic exposure to glycerol

Alternative techniques for the cryopreservation of kangaroo spermatozoa that reduced or eliminated the need for glycerol were investigated including; (1) freezing spermatozoa with 20% glycerol in pre-packaged 0.25 mL Cassou straws to enable rapid dilution of the glycerol post-thaw, (2) investigating the efficacy of 20% (v/v) dimethyl sulphoxide (DMSO) and dimethylacetamide (DMA—10%, 15% and 20% v/v) as cryoprotectants and (3) vitrification of spermatozoa with or without cryoprotectant (20% v/v glycerol, 20% v/v DMSO and 20% v/v DMA). Immediate in-straw post-thaw dilution of 20% glycerol and cryopreservation of spermatozoa in 20% DMSO produced no significant improvement in post-thaw viability of kangaroo spermatozoa. Spermatozoa frozen in 20% DMA showed post-thaw motility and plasma membrane integrity of 12.7 ± 1.9% and 22.7 ± 5.4%, respectively, while kangaroo spermatozoa frozen by ultra-rapid freezing techniques showed no evidence of post-thaw viability. The use of 10–20% DMA represents a modest but significant improvement in the development of a sperm cryopreservation procedure for kangaroos.     

Comparison of marine spatial planning methods in Madagascar demonstrates value of alternative approaches

The Government of Madagascar plans to increase marine protected area coverage by over one million hectares. To assist this process, we compare four methods for marine spatial planning of Madagascar's west coast. Input data for each method was drawn from the same variables: fishing pressure, exposure to climate change, and biodiversity (habitats, species distributions, biological richness, and biodiversity value). The first method compares visual color classifications of primary variables, the second uses binary combinations of these variables to produce a categorical classification of management actions, the third is a target-based optimization using Marxan, and the fourth is conservation ranking with Zonation. We present results from each method, and compare the latter three approaches for spatial coverage, biodiversity representation, fishing cost and persistence probability. All results included large areas in the north, central, and southern parts of western Madagascar. Achieving 30% representation targets with Marxan required twice the fish catch loss than the categorical method. The categorical classification and Zonation do not consider targets for conservation features. However, when we reduced Marxan targets to 16.3%, matching the representation level of the "strict protection" class of the categorical result, the methods show similar catch losses. The management category portfolio has complete coverage, and presents several management recommendations including strict protection. Zonation produces rapid conservation rankings across large, diverse datasets. Marxan is useful for identifying strict protected areas that meet representation targets, and minimize exposure probabilities for conservation features at low economic cost. We show that methods based on Zonation and a simple combination of variables can produce results comparable to Marxan for species representation and catch losses, demonstrating the value of comparing alternative approaches during initial stages of the planning process. Choosing an appropriate approach ultimately depends on scientific and political factors including representation targets, likelihood of adoption, and persistence goals.     

Linked thioredoxin-glutathione systems in platyhelminth parasites: alternative pathways for glutathione reduction and deglutathionylation

In most organisms, thioredoxin (Trx) and/or glutathione (GSH) systems are essential for redox homeostasis and deoxyribonucleotide synthesis. Platyhelminth parasites have a unique and simplified thiol-based redox system, in which the selenoprotein thioredoxin-glutathione reductase (TGR), a fusion of a glutaredoxin (Grx) domain to canonical thioredoxin reductase domains, is the sole enzyme supplying electrons to oxidized glutathione (GSSG) and Trx. This enzyme has recently been validated as a key drug target for flatworm infections. In this study, we show that TGR possesses GSH-independent deglutathionylase activity on a glutathionylated peptide. Furthermore, we demonstrate that deglutathionylation and GSSG reduction are mediated by the Grx domain by a monothiolic mechanism and that the glutathionylated TGR intermediate is resolved by selenocysteine. Deglutathionylation and GSSG reduction via Grx domain, but not Trx reduction, are inhibited at high [GSSG]/[GSH] ratios. We found that Trxs (cytosolic and mitochondrial) provide alternative pathways for deglutathionylation and GSSG reduction. These pathways are operative at high [GSSG]/[GSH] and function in a complementary manner to the Grx domain-dependent one. Despite the existence of alternative pathways, the thioredoxin reductase domains of TGR are an obligate electron route for both the Grx domain- and the Trx-dependent pathways. Overall, our results provide an explanation for the unique array of thiol-dependent redox pathways present in parasitic platyhelminths. Finally, we found that TGR is inhibited by 1-hydroxy-2-oxo-3-(N-3-methyl-aminopropyl)-3-methyl-1-triazene (NOC-7), giving further evidence for NO donation as a mechanism of action for oxadiazole N-oxide TGR inhibitors. Thus, NO donors aimed at TGR could disrupt the entire redox homeostasis of parasitic flatworms.     

Parasite histories and novel phylogenetic tools: alternative approaches to inferring parasite evolution from molecular markers

Parasitological research is often contingent on the knowledge of the phylogeny/genealogy of the studied group. Although molecular phylogenetics has proved to be a powerful tool in such investigations, its application in the traditional fashion, based on a tree inference from the primary nucleotide sequences may, in many cases, be insufficient or even improper. These limitations are due to a number of factors, such as a scarcity/ambiguity of phylogenetic information in the sequences, an intricacy of gene relationships at low phylogenetic levels, or a lack of criteria when deciding among several competing coevolutionary scenarios. With respect to the importance of a precise and reliable phylogenetic background in many biological studies, attempts are being made to extend molecular phylogenetics with a variety of new data sources and methodologies. In this review, selected approaches potentially applicable to parasitological research are presented and their advantages as well as drawbacks are discussed. These issues include the usage of idiosyncratic markers (unique features with presumably low probability of homoplasy), such as insertion of mobile elements, gene rearrangements and secondary structure features; the problem of ancestral polymorphism and reticulate relationships at low phylogenetic levels; and the utility of a molecular clock to facilitate discrimination among alternative scenarios in host-parasite coevolution.