Physiological stress in decapod crustaceans (Munida rugosa and Liocarcinus depurator) discarded in the Clyde Nephrops fishery

Crustacean discards experience stress during commercial fishing operations, due to increased exercise while in the trawl and aerial exposure during sorting of the catch. Physiological stress and recovery were assessed following trawling of two ecologically important decapod species, regularly discarded in the Clyde Nephrops fishery. Haemolymph samples taken from trawled swimming crabs, Liocarcinus depurator, and squat lobsters, Munida rugosa, had significantly higher concentrations of ammonia (0.308 and 0.519 mmol l(-1)), D-glucose (0.14 and 0.097 mmol l(-1)) and L-lactate (6.2 and 0.87 mmol l(-1)) compared with controls, indicating an impairment of ammonia excretion and a switch to anaerobic metabolism. Concurrently, the haemolymph pH of trawled squat lobsters was low (7.47) compared with controls (7.75); however, the reverse trend was found in L. depurator. Initially elevated lactate (7.98 mmol l(-1)) and glucose (0.73 mmol l(-1)) concentrations of trawled and emersed (1 h) L. depurator were restored, 4 h after re-immersion along with pH (7.54). Crabs that had been emersed for 1 h had significantly higher concentrations of glucose (0.2 mmol l(-1)) and lactate (5.14 mmol l(-1)), and had more acidic blood (7.64) than L. depurator subject to 1 h of exercise, indicating that anoxia was the main cause of physiological stress. Crabs and squat lobsters lost 7% and 9% of their initial body wet weight following 1 h of emersion, although blood osmolarities did not change significantly. While all animals survived aerial exposure in our experiments, sorting of the catch on commercial boats takes up to 300 min, which could lead to mortality or sub-lethal chronic biochemical changes that could compromise fitness.     

A Reciprocal Relationship Between the Genetic Diversity at Two Metabolically-linked Isozyme Loci in Several Conifer Species

In the context of a general survey on genetic variation of isozyme-gene systems which function in the carbohydrate degradation and conversion, we detected a reciprocal relationship between genetic diversity at the hexokinase (HEK-A) and phosphoglucose isomerase (PGI-B) loci in Scots pine populations. Further studies on Norway spruce, Douglas-fir and Siberian stone pine revealed that this relationship appears to be a more general phenomenon in conifers such that increasing diversity at one locus is correlated with a decrease in diversity at the other locus. Since the two gene loci are not structurally linked but are encoding enzymes of two sucessive metabolic steps in the glucose conversion towards glycolysis, it is assumed that some sort of selection, especially during germination and early embryo development, may be the causal explanation. A metabolically-based model incorporating selective advantage and disadvantage of alternate two-locus genotypes at HEK-A/PGI-B was presented in order to elucidate the possible adaptive nature of this reciprocal relationship. This revised version was published online in July 2006 with corrections to the Cover Date.     

Determination of crucial epitopes in the sperm protein calsperin employing synthetic peptides and monoclonal antibodies

The chaperone protein calsperin is exclusively expressed in the testes and is essential for sperm migration from the uterus into the oviduct. During spermatogenesis, calsperin interacts with ADAM3, a spermatozoon membrane protein required for fertilization. In this study, we characterized a calsperin epitope by using two monoclonal antibodies and resin-bound calsperin peptides, which were tested for reactivity using a modified enzyme-linked immunosorbent assay. An epitope located at the C-terminal end of calsperin corresponding to amino acids ²²⁸WEKHFLDAS²³⁷ was identified. Three hot spot amino acids were essential for antibody binding whereas the remaining amino acids in the identified epitope appeared to be essential for bringing the critical contact residues into an α-helix structure. No notable sequence similarity was determined between the identified calsperin epitope and calreticulin, a chaperone homologue with sequence similarity, indicating that the identified epitope was specific for calsperin. Characterization of the calsperin epitope and of the two antibodies tested may be used in assays for further characterization of calsperin, where knowledge about the binding sites is necessary, for example, in sandwich assays. Moreover, studies like these may be used to study the function of calsperin during spermatogenesis and fertilization in detail and to develop new male contraception methods by targeting calsperin and mediating neutralization of its function.     

Widespread seafloor anoxia during generation of the Ediacaran Shuram carbon isotope excursion

Reconstructing the oxygenation history of Earth's oceans during the Ediacaran period (635 to 539 million years ago) has been challenging, and this has led to a polarizing debate about the environmental conditions that played host to the rise of animals. One focal point of this debate is the largest negative inorganic C-isotope excursion recognized in the geologic record, the Shuram excursion, and whether this relic tracks the global-scale oxygenation of Earth's deep oceans. To help inform this debate, we conducted a detailed geochemical investigation of two siliciclastic-dominated successions from Oman deposited through the Shuram Formation. Iron speciation data from both successions indicate formation beneath an intermittently anoxic local water column. Authigenic thallium (Tl) isotopic compositions leached from both successions are indistinguishable from bulk upper continental crust (ε²⁰⁵Tl_A ≈ −2) and, by analogy with modern equivalents, likely representative of the ancient seawater ε²⁰⁵Tl value. A crustal seawater ε²⁰⁵Tl value requires limited manganese (Mn) oxide burial on the ancient seafloor, and by extension widely distributed anoxic sediment porewaters. This inference is supported by muted redox-sensitive element enrichments (V, Mo, and U) and consistent with some combination of widespread (a) bottom water anoxia and (b) high sedimentary organic matter loading. Contrary to a classical hypothesis, our interpretations place the Shuram excursion, and any coeval animal evolutionary events, in a predominantly anoxic global ocean.     

Ifit2 Deficiency Results in Uncontrolled Neurotropic Coronavirus Replication and Enhanced Encephalitis via Impaired Alpha/Beta Interferon Induction in Macrophages

Type I interferons (IFN-α/β) limit viral dissemination prior to the emergence of adaptive immune responses through the concerted action of interferon-stimulated genes (ISGs). Although IFN-α/β induction by coronaviruses is modest, it effectively limits viral spread within the central nervous system (CNS) and protects against mortality. The protective roles of specific ISGs against the mouse hepatitis virus (MHV) members of the coronaviruses are largely unknown. This study demonstrates a protective role of the ISG Ifit2 in encephalitis induced by the dual hepato- and neurotropic MHV-A59. Contrasting the mild encephalitis and 100% survival of MHV-A59-infected wild-type (wt) mice, nearly 60% of infected Ifit2^−/− mice exhibited severe encephalitis and succumbed between 6 and 8 days postinfection. Increased clinical disease in Ifit2^−/− mice coincided with higher viral loads and enhanced viral spread throughout the CNS parenchyma. Ifit2^−/− mice also expressed significantly reduced IFN-α/β and downstream ISG mRNAs Ifit1, Isg15, and Pkr, while expression of proinflammatory cytokines and chemokines was only modestly affected in the CNS. Impaired IFN-α/β induction in the absence of Ifit2 was confirmed by ex vivo mRNA analysis of microglia and macrophages, the prominent cell types producing IFN-α/β following MHV CNS infection. Furthermore, both IFN-α/β mRNA and protein production were significantly reduced in MHV-infected Ifit2^−/− relative to wt bone marrow-derived macrophages. Collectively, the data implicate Ifit2 as a positive regulator of IFN-α/β expression, rather than direct antiviral mediator, during MHV-induced encephalitis.     

Determination of typical patterns from strongly varying signals

Forces measured in human joints vary considerably when an activity such as walking is carried out by different subjects or when it is repeated. 'Typical' standardised force-time patterns are needed to test and improve joint implants. Mechanically most important for their endurance are the magnitudes and times of force maxima and minima. They should equal the arithmetic means from the single measurements. Similar problems exist when evaluating other strongly varying signals, as in gait analysis. The new method to calculate typical signals (TSs) enhances existing dynamic time warping (DTW) procedures. It allows us to combine any number of signals. The sequence of input signals--used for calculating the TS--has only a minor influence. The accuracy of the method was tested numerically on signals for which the typical patterns could be defined exactly, and also on real joint forces that varied to different extents.     

Imbalanced Oxidative Stress Causes Chlamydial Persistence during Non-Productive Human Herpes Virus Co-Infection

Both human herpes viruses and Chlamydia are highly prevalent in the human population and are detected together in different human disorders. Here, we demonstrate that co-infection with human herpes virus 6 (HHV6) interferes with the developmental cycle of C. trachomatis and induces persistence. Induction of chlamydial persistence by HHV6 is independent of productive virus infection, but requires the interaction and uptake of the virus by the host cell. On the other hand, viral uptake is strongly promoted under co-infection conditions. Host cell glutathione reductase activity was suppressed by HHV6 causing NADPH accumulation, decreased formation of reduced glutathione and increased oxidative stress. Prevention of oxidative stress restored infectivity of Chlamydia after HHV6-induced persistence. We show that co-infection with Herpes simplex virus 1 or human Cytomegalovirus also induces chlamydial persistence by a similar mechanism suggesting that Chlamydia -human herpes virus co-infections are evolutionary shaped interactions with a thus far unrecognized broad significance.     

The high prevalence of Torque teno virus DNA in blood donors and haemodialysis patients in southern Brazil

This study investigates the frequency of Torque teno virus (TTV) infection in 150 blood donors and 77 patients requiring haemodialysis in southern Brazil. Plasma samples were screened for TTV DNA using polymerase chain reaction (PCR). The prevalences of TTV among blood donors and patients requiring haemodialysis were 73.3% and 68.8%, respectively. The presence of TTV was correlated with age in the blood donors (p = 0.024). In haemodialysis patients, no association was found between TTV infection and the demographic parameters (age, sex and education), the duration of haemodialysis or a history of blood transfusion. This study is the first to evaluate the prevalence of TTV infection in Brazilian patients requiring haemodialysis.