Recurrent warming to improve cold storage of Trichogrammatids (Hymenoptera: Trichogrammatidae)

Cold storage can be used to slow development, facilitate accumulation of the organisms and accommodate fluctuating demand for augmentative biological control agents. Previous research suggested the possibility of improving cold storage of Trichogrammatids by recurrent warming, so we subjected Trichogramma ostriniae juveniles within Ephestia kuehniella host eggs to either 2°C constant or to 2°C with twice-weekly recurrent warming for 3 h to 20°C. Parasitoid subsamples were allowed to mature for 1–9 days before placement in cold storage for up to 8 weeks. Parasitism by parentals, progeny emergence and fecundity and longevity of progeny were measured weekly for 8 weeks. Relative to constant 2°C, recurrent warming generally improved emergence, fecundity and longevity, and all the response variables were affected by the interaction of temperature regimen, parasitoid maturity class, and cold storage duration. This implies the utility of recurrent warming to improve egg parasitoid performance and for extending the duration of cold storage.     

Dispersal of Trichogramma ostriniae in field corn

Trichogramma ostriniae has shown success as a biological control agent for European corn borer (Ostrinia nubilalis) in sweet corn and the species offers potential for suppression of lepidopteran pests of field corn. Field corn is typically planted at higher densities, is taller, and has greater leaf area than sweet corn, presenting a possible restriction on T. ostriniae dispersal and efficacy. Therefore, parasitoid dispersal in field corn from the centre of a 6.25 ha square grid was determined using sticky cards to capture adult T. ostriniae and sentinel eggs of O. nubilalis to monitor parasitism after releases of ~1 million of T. ostriniae each into four fields of corn. Dispersal was rapid and extensive, achieving distances of ~175 m within 4–7 days after release. The pattern of movement fit well with a diffusion model of dispersal, with the greatest level of dispersal occurring from 7 to 10 days post-release. Parasitism of O. nubilalis sentinel egg masses declined linearly with distance from the release foci, and was also greatest 7–10 days post-release. However, measurement of association showed no significant differences between the spatial distributions of sticky trap captures and percentage parasitism of O. nubilalis egg masses. The distances from the release point that encompassed 98% of re-captured T. ostriniae increased over time and were estimated to range from a low of 100 m at 4 days post-release to 365 m at 14 days post-release. The results of this research suggest that T. ostriniae relies initially on random movement to locate host patches, and that a single release locus per hectare would be sufficient in field corn.     

Evolutionary and functional conservation of the DNA non-homologous end-joining protein, XLF/Cernunnos

Non-homologous end-joining is a major pathway of DNA double-strand break repair in mammalian cells, deficiency in which confers radiosensitivity and immune deficiency at the whole organism level. A core protein complex comprising the Ku70/80 heterodimer together with a complex between DNA ligase IV and XRCC4 is conserved throughout eukaryotes and assembles at double-strand breaks to mediate ligation of broken DNA ends. In Saccharomyces cerevisiae an additional NHEJ protein, Nej1p, physically interacts with the ligase IV complex and is required in vivo for ligation of DNA double-strand breaks. Recent studies with cells derived from radiosensitive and immune-deficient patients have identified the human protein, XLF (also named Cernunnos), as a crucial NHEJ protein. Here we show that XLF and Nej1p are members of the same protein superfamily and that this family has members in diverse eukaryotes. Indeed, we show that a member of this family encoded by a previously uncharacterized open-reading frame in the Schizosaccharomyces pombe genome is required for NHEJ in this organism. Furthermore, our data reveal that XLF family proteins can bind to DNA and directly interact with the ligase IV-XRCC4 complex to promote DSB ligation. We therefore conclude that XLF family proteins interact with the ligase IV-XRCC4 complex to constitute the evolutionarily conserved enzymatic core of the NHEJ machinery.     

Cooperative regulation of extracellular signal-regulated kinase activation and cell shape change by filamin A and beta-arrestins

beta-Arrestins (betaarr) are multifunctional adaptor proteins that can act as scaffolds for G protein-coupled receptor activation of mitogen-activated protein kinases (MAPK). Here, we identify the actin-binding and scaffolding protein filamin A (FLNA) as a betaarr-binding partner using Son of sevenless recruitment system screening, a classical yeast two-hybrid system, coimmunoprecipitation analyses, and direct binding in vitro. In FLNA, the betaarr-binding site involves tandem repeat 22 in the carboxyl terminus. betaarr binds FLNA through both its N- and C-terminal domains, indicating the presence of multiple binding sites. We demonstrate that betaarr and FLNA act cooperatively to activate the MAPK extracellular signal-regulated kinase (ERK) downstream of activated muscarinic M1 (M1MR) and angiotensin II type 1a (AT1AR) receptors and provide experimental evidence indicating that this phenomenon is due to the facilitation of betaarr-ERK2 complex formation by FLNA. In Hep2 cells, stimulation of M1MR or AT1AR results in the colocalization of receptor, betaarr, FLNA, and active ERK in membrane ruffles. Reduction of endogenous levels of betaarr or FLNA and a catalytically inactive dominant negative MEK1, which prevents ERK activation, inhibit membrane ruffle formation, indicating the functional requirement for betaarr, FLNA, and active ERK in this process. Our results indicate that betaarr and FLNA cooperate to regulate ERK activation and actin cytoskeleton reorganization.     

Atlantic Bluefin Tuna in the Gulf of Maine, I: Estimation of Seasonal Abundance Accounting for Movement, School and School-Aggregation Behaviour

Direct assessment of the abundance of highly migratory pelagic species, such as tuna, is rarely available and most indices are based on catch information. We estimate the seasonal abundance of North Atlantic bluefin tuna, Thunnus thynnus, in the Gulf of Maine (GOM) from a 3-year aerial survey conducted with commercial spotter pilots, while also utilizing findings from analyses of tracking and tagging data. We apply statistical correction and calibration to seasonal abundance estimates accounting for measured changes in horizontal and vertical movement behaviour, size, shape and aggregation of bluefin tuna schools. Our approach relies on ecological knowledge of bluefin tuna to extrapolate survey observations across areas not sampled by correcting survey abundance estimates based on range of movement search pattern and depth preference. We demonstrate how separate findings obtained through the analysis of data collected across different spatial and temporal scales can be integrated to correct and calibrate estimates of population abundance. We obtain fitted estimates of seasonal abundance of bluefin tuna in the GOM during 1994–1996 in the range of 45,000–51,000 individuals. If tuna behaviour is not accounted for, we estimate that the base or residual survey precision would be 4–7% determined from analysis of recent spotter survey data in the study region. We estimate the precision in estimating seasonal abundance accounting for tuna behaviour to lie within a range of 1,301–3,302%. Under hypothetical future improvements in survey design that achieve a precision of 20% in transect length and placement, we calculate net-precision to lie within a range of 82–93%. This calculation assumes reducible uncertainty in school size estimation and irreducible uncertainty in movement and school-aggregation behaviour. We infer that survey precision could be further reduced by 43–32% to attain 10–50% in which a 3–8 years adaptive survey design may reliably detect a seasonal abundance trend.     

Sperm design and function in the redside dace Clinostomus elongatus

A study was undertaken to examine sperm morphometry in relation to sperm velocity and sperm longevity in the redside dace Clinostomus elongatus . There was significant between-male variance in sperm size and shape metrics (total sperm length, sperm head length, flagellum length and sperm head length to width ratio) and positive relationships were found between these morphometrics and sperm velocity. There were no significant relationships found between sperm morphometry and sperm longevity, nor was there a trade-off between sperm velocity and sperm longevity.     

Secondary sexual characters and sperm traits in coho salmon Oncorhynchus kisutch

A study was undertaken to examine secondary sexual characters (spawning colouration and overall body size) in relation to sperm metrics in one alternative reproductive tactic of coho salmon Oncorhynchus kisutch : large hooknose males that spawn in dominance-based hierarchies. Males with less intense red spawning colouration had higher sperm velocities than males with darker red spawning colouration. There was no relationship between male body size and sperm metrics. These results suggest that within an alternative reproductive tactic, variation in sperm competition intensity may select for a trade-off between investment in sexual colouration and sperm quality.     

Proteomic analysis of secreted membrane vesicles of archaeal Sulfolobus species reveals the presence of endosome sorting complex components

The crenarchaea Sulfolobus acidocaldarius, S. solfataricus and S. tokodaii, release membrane vesicles into the medium. These membrane vesicles consist of tetraether lipids and are coated with an S-layer. A proteomic analysis reveals the presence of proteins homologous to subunits of the eukaryotic endosomal sorting complex required for transport (ESCRT). Immunodetection of one of these homologs suggest a cell surface localization in intact cells. These data suggest that the membrane vesicles in Sulfolobus sp. emerge from a specific budding process with similarity to the endosomal sorting pathway.     

Pesticide exposure: the hormonal function of the female reproductive system disrupted?

Some pesticides may interfere with the female hormonal function, which may lead to negative effects on the reproductive system through disruption of the hormonal balance necessary for proper functioning. Previous studies primarily focused on interference with the estrogen and/or androgen receptor, but the hormonal function may be disrupted in many more ways through pesticide exposure. The aim of this review is to give an overview of the various ways in which pesticides may disrupt the hormonal function of the female reproductive system and in particular the ovarian cycle. Disruption can occur in all stages of hormonal regulation: 1. hormone synthesis; 2. hormone release and storage; 3. hormone transport and clearance; 4. hormone receptor recognition and binding; 5. hormone postreceptor activation; 6. the thyroid function; and 7. the central nervous system. These mechanisms are described for effects of pesticide exposure in vitro and on experimental animals in vivo. For the latter, potential effects of endocrine disrupting pesticides on the female reproductive system, i.e. modulation of hormone concentrations, ovarian cycle irregularities, and impaired fertility, are also reviewed. In epidemiological studies, exposure to pesticides has been associated with menstrual cycle disturbances, reduced fertility, prolonged time-to-pregnancy, spontaneous abortion, stillbirths, and developmental defects, which may or may not be due to disruption of the female hormonal function. Because pesticides comprise a large number of distinct substances with dissimilar structures and diverse toxicity, it is most likely that several of the above-mentioned mechanisms are involved in the pathophysiological pathways explaining the role of pesticide exposure in ovarian cycle disturbances, ultimately leading to fertility problems and other reproductive effects. In future research, information on the ways in which pesticides may disrupt the hormonal function as described in this review, can be used to generate specific hypotheses for studies on the effects of pesticides on the ovarian cycle, both in toxicological and epidemiological settings.     

G protein-coupled receptor kinase 6A phosphorylates the Na(+)/H(+) exchanger regulatory factor via a PDZ domain-mediated interaction.

The Na(+)/H(+) exchanger regulatory factor (NHERF) is constitutively phosphorylated in cells, but the site(s) of this phosphorylation and the kinase(s) responsible for it have not been identified. We show here that the primary site of constitutive NHERF phosphorylation in human embryonic kidney 293 (HEK-293) cells is Ser(289), and that the stoichiometry of phosphorylation is near 1 mol/mol. NHERF contains two PDZ domains that recognize the sequence S/T-X-L at the carboxyl terminus of target proteins, and thus we examined the possibility that kinases containing this motif might associate with and phosphorylate NHERF. Overlay experiments and co-immunoprecipitation studies revealed that NHERF binds with high affinity to a splice variant of the G protein-coupled receptor kinase 6, GRK6A, which terminates in the motif T-R-L. NHERF does not associate with GRK6B or GRK6C, alternatively spliced variants that differ from GRK6A at their extreme carboxyl termini. GRK6A phosphorylates NHERF efficiently on Ser(289) in vitro, whereas GRK6B, GRK6C, and GRK2 do not. Furthermore, the endogenous "NHERF kinase" activity in HEK-293 cell lysates is sensitive to treatments that alter the activity of GRK6A. These data suggest that GRK6A phosphorylates NHERF via a PDZ domain-mediated interaction and that GRK6A is the kinase in HEK-293 cells responsible for the constitutive phosphorylation of NHERF.