Structural changes caused by thyrotropin in thyroid cells and in liposomes containing reconstituted thyrotropin receptor

Thyrotropin causes a rapid and significant increase in the fluorescence polarization of DPH when this hydrophobic probe is incorporated into a strain of functioning rat thyroid cells (FRTL5). This increase is ligand-specific and is not related to cAMP production. The phenomenon seems to reflect the interaction of thyrotropin with the glycoprotein component of its membrane receptor, as suggested by experiments in which thyrotropin causes increases in DPH fluorescence polarization in liposomes embedded with this receptor component but not with gangliosides. A strain of nonfunctioning rat thyroid cells (FRT), exhibiting no reactivity with monoclonal antibodies to the glycoprotein component of the thyrotropin receptor, requires two orders of magnitude higher concentrations of thyrotropin to exhibit a comparable phenomenon.     

Molecular drift of the bride of sevenless (boss) gene in Drosophila

DNA sequences were determined for three to five alleles of the bride-of- sevenless (boss) gene in each of four species of Drosophila. The product of boss is a transmembrane receptor for a ligand coded by the sevenless gene that triggers differentiation of the R7 photoreceptor cell in the compound eye. Population parameters affecting the rate and pattern of molecular evolution of boss were estimated from the multinomial configurations of nucleotide polymorphisms of synonymous codons. The time of divergence between D. melanogaster and D. simulans was estimated as approximately 1 Myr, that between D. teissieri and D. yakuba as approximately 0.75 Myr, and that between the two pairs of sibling species as approximately 2 Myr. (The boss genes themselves have estimated divergence times approximately 50% greater than the species divergence times.) The effective size of the species was estimated as approximately 5 x 10(6), and the average mutation rate was estimated as 1-2 x 10(-9)/nucleotide/generation. The ratio of amino acid polymorphisms within species to fixed differences between species suggests that approximately 25% of all possible single-step amino acid replacements in the boss gene product may be selectively neutral or nearly neutral. The data also imply that random genetic drift has been responsible for virtually all of the observed differences in the portion of the boss gene analyzed among the four species.      

The contractile basis of amoeboid movement: V. The control of gelation, solation, and contraction in extracts from dictyostelium discoideum

Motile extracts have been prepared from Dictyostelium discoideum by homogenization and differential centrifugation at 4 degrees C in a stabilization solution (60). These extracts gelled on warming to 25 degrees Celsius and contracted in response to micromolar Ca++ or a pH in excess of 7.0. Optimal gelation occurred in a solution containing 2.5 mM ethylene glycol-bis (β-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA), 2.5 mM piperazine-N-N'-bis [2-ethane sulfonic acid] (PIPES), 1 mM MgC1(2), 1 mM ATP, and 20 mM KCI at ph 7.0 (relaxation solution), while micromolar levels of Ca++ inhibited gelation. Conditions that solated the gel elicited contraction of extracts containing myosin. This was true regardless of whether chemical (micromolar Ca++, pH >7.0, cytochalasin B, elevated concentrations of KCI, MgC1(2), and sucrose) or physical (pressure, mechanical stress, and cold) means were used to induce solation. Myosin was definitely required for contraction. During Ca++-or pH-elicited contraction: (a) actin, myosin, and a 95,000-dalton polypeptide were concentrated in the contracted extract; (b) the gelation activity was recovered in the material sqeezed out the contracting extract;(c) electron microscopy demonstrated that the number of free, recognizable F-actin filaments increased; (d) the actomyosin MgATPase activity was stimulated by 4- to 10-fold. In the absense of myosin the Dictyostelium extract did not contract, while gelation proceeded normally. During solation of the gel in the absense of myosin: (a) electron microscopy demonstrated that the number of free, recognizable F- actin filaments increased; (b) solation-dependent contraction of the extract and the Ca++-stimulated MgATPase activity were reconstituted by adding puried Dictyostelium myosin. Actin purified from the Dictyostelium extract did not gel (at 2 mg/ml), while low concentrations of actin (0.7-2 mg/ml) that contained several contaminating components underwent rapid Ca++ regulated gelation. These results indicated : (a) gelation in Dictyostelium extracts involves a specific Ca++-sensitive interaction between actin and several other components; (b) myosin is an absolute requirement for contraction of the extract; (c) actin-myosin interactions capable of producing force for movement are prevented in the gel, while solation of the gel by either physical or chemical means results in the release of F-actin capable of interaction with myosin and subsequent contraction. The effectiveness of physical agents in producting contraction suggests that the regulation of contraction by the gel is structural in nature.     

Variation of annual apparent survival and detection rates with age,year and individual identity in male Weddell seals (Leptonychotes weddellii) from long-term mark-recapture data

Exploring age- and sex-specific survival rates provides insight regarding population behavior and life-history trait evolution. However, our understanding of how age-specific patterns of survival, including actuarial senescence, compare between the sexes remains inadequate. Using 36 years of mark-recapture data for 7,516 male Weddell seals (Leptonychotes weddellii) born in Erebus Bay, Antarctica, we estimated age-specific annual survival rates using a hierarchical model for mark-recapture data in a Bayesian framework. Our male survival estimates were moderate for pups and yearlings, highest for 2-year-olds, and gradually declined with age thereafter such that the oldest animals observed had the lowest rates of any age. Reports of senescence in other wildlife populations of species with similar longevity occurred at older ages than those presented here. When compared to recently published estimates for reproductive Weddell seal females, we found that peak survival rates were similar (males: 0.94, 95% CI = 0.92–0.96; females: 0.92, 95% CI = 0.93–0.95), but survival rates at older ages were lower in males. Age-specific male Weddell seal survival rates varied across years and individuals, with greater variation occurring across years. Similar studies on a broad range of species are needed to contextualize these results for a better understanding of the variation in senescence patterns between the sexes of the same species, but our study adds information for a marine mammal species to a research topic dominated by avian and ungulate species.     

Nasal Discs and the Vital Rates of Lesser Scaup

Nasal discs have been used to identify ducks in studies of survival and reproduction. To date, there has not been a comprehensive assessment of nasal-disc effects on the vital rates of wild ducks. We applied nasal discs to 603 juvenile and 784 adult lesser scaup (Aythya affinis) females from a population breeding in southwest Montana, USA, and released 1,399 juvenile and 71 adult females wearing only metal leg bands between June 2005 and September 2016. Using resighting, recapture, and hunter-recovery data collected from those individuals, we estimated survival and recovery probability with multistate capture-recapture models in Program MARK. We also assessed if recovery distance from our study site and pre-breeding and brood-rearing body condition were diminished for females wearing nasal discs. Model-averaged survival probabilities were 0.231 ± 0.035 (SE) for juveniles and 0.482 ± 0.019 for adults released with nasal discs. Survival was 1.8–3.4 times higher for females released with metal leg bands when compared to those released with nasal discs; survival of these juveniles was 0.433 ± 0.049 and 0.693 ± 0.039 for adults. We did not find evidence for recovery probability or recovery distance varying between females that wore nasal discs and those that did not. During the pre-breeding and brood-rearing seasons, we did not find females wearing nasal discs to be in lower body condition when compared to unmarked females. Our comprehensive assessment of nasal discs on wild lesser scaup suggests that survival probabilities estimated from nasal-marked study populations should be cautiously interpreted as minimum estimates. © 2021 The Wildlife Society.     

Variability in temporary emigration rates of individually marked female Weddell seals prior to first reproduction

In many species, temporary emigration (TE) is a phenomenon, often indicative of life-history characteristics such as dormancy, skipped reproduction, or partial migration, whereby certain individuals in a population are temporarily unobservable at a particular site. TE may be a flexible condition-dependent strategy that allows individuals to mitigate effects of adverse conditions. Consequently, TE rates ought to be highly variable, but sources of variations are poorly understood for most species. We used data from known-aged female Weddell seals (Leptonychotes weddellii) tagged in Erebus Bay, Antarctica, to investigate sources of variation in TE rates prior to reproduction and to evaluate possible implications for age-specific probability of first reproduction. TE rates were near 1 the year after birth, decreased to an average of 0.15 ( $ \widehat{\text{SE}} $  = 0.01) by age 8, and were similar thereafter. TE rates varied substantially from year-to-year and were lower for seals that attended reproductive colonies the previous year than for seals that did not attend (e.g., $ \overline{{\hat{\psi }_{{i,{\text{age}}\,8}}^{\text{UU}} - \hat{\psi }_{{i,\,{\text{age}}\,8}}^{\text{PU}} }} $  = 0.22). Recruitment rates were marginally greater for seals that did attend than for seals that did not attend colonies the previous year. For Weddell seals specifically, our results suggest that (1) motivation to attend colonies varied temporally, (2) as seals grew older they had increased motivation to attend even before reproductive maturity, and (3) seals appear to follow various attendance strategies. More broadly, our results support the idea of TE as a variable, condition-dependent strategy, and highlight the utility of TE models for providing population and life-history insights for diverse taxa.     

Distribution of duck broods relative to habitat characteristics in the Prairie Pothole Region

Conservation programs for breeding ducks in the Prairie Pothole Region (PPR) of the United States and Canada require effective means of evaluating and characterizing breeding habitat across large landscapes. Extensive surveys of the distribution of duck broods in late-summer could help identify wetland basins with greater probabilities of occupancy. Broods are difficult to detect, however, rendering presence–absence data from single-visit surveys difficult to interpret, particularly when probability of detection is related to habitat features. Multiple-visit occupancy surveys offer a potential solution. From 20 July to 5 August 2007–2009, we conducted a 3-visit survey of wetland basins located on 167 10.4-km² study plots in the PPR. Our survey focused on broods of the 5 most common breeding duck species (Anas spp.). Our main objectives were to investigate ecological relationships between occupancy of wetland basins by broods and habitat characteristics and to examine if habitat-specific detection was of enough concern to warrant multi-survey approaches in the future. We surveyed 3,226 wetland basins during the study. Probability of occupancy of a wetland basin by a brood was positively related to the log of wet area for all 5 study species and was greater on wetlands located on plots with a greater proportion of herbaceous perennial cover for 4 of 5 species. For example, the median probability of occupancy for gadwall (Anas strepera) increased from 0.08 (90% Credible Interval [CrI]: 0.07, 0.10) to 0.28 (90% CrI: 0.23, 0.33) as wet area increased from 0.19 ha to 2.12 ha, and increased from 0.12 (90% CrI: 0.09, 0.16) to 0.20 (90% CrI: 0.16, 0.25) as proportion of perennial grass cover on the study plot increased from 0.03 to 0.99. Because occupancy and detection were both related to attributes of wetland basins, we concluded that the multiple-visit survey was a useful approach for identifying habitat relationships of duck broods. Our results indicated that most broods of the study species were found in 10.4-km² landscapes with greater densities of small- to mid-sized wetland basins and a greater proportion of herbaceous perennial vegetation. Our study provided new empirical support that could be used to help target conservation actions to the most productive landscapes for breeding ducks. © 2012 The Wildlife Society.     

Saikosaponin-d,a novel SERCA inhibitor,induces autophagic cell death in apoptosis-defective cells

Autophagy is an important cellular process that controls cells in a normal homeostatic state by recycling nutrients to maintain cellular energy levels for cell survival via the turnover of proteins and damaged organelles. However, persistent activation of autophagy can lead to excessive depletion of cellular organelles and essential proteins, leading to caspase-independent autophagic cell death. As such, inducing cell death through this autophagic mechanism could be an alternative approach to the treatment of cancers. Recently, we have identified a novel autophagic inducer, saikosaponin-d (Ssd), from a medicinal plant that induces autophagy in various types of cancer cells through the formation of autophagosomes as measured by GFP-LC3 puncta formation. By computational virtual docking analysis, biochemical assays and advanced live-cell imaging techniques, Ssd was shown to increase cytosolic calcium level via direct inhibition of sarcoplasmic/endoplasmic reticulum Ca²⁺ ATPase pump, leading to autophagy induction through the activation of the Ca²⁺/calmodulin-dependent kinase kinase–AMP-activated protein kinase–mammalian target of rapamycin pathway. In addition, Ssd treatment causes the disruption of calcium homeostasis, which induces endoplasmic reticulum stress as well as the unfolded protein responses pathway. Ssd also proved to be a potent cytotoxic agent in apoptosis-defective or apoptosis-resistant mouse embryonic fibroblast cells, which either lack caspases 3, 7 or 8 or had the Bax-Bak double knockout. These results provide a detailed understanding of the mechanism of action of Ssd, as a novel autophagic inducer, which has the potential of being developed into an anti-cancer agent for targeting apoptosis-resistant cancer cells.     

Drivers of <Emphasis Type="Italic">Bromus tectorum</Emphasis> Abundance in the Western North American Sagebrush Steppe

Bromus tectorum can transform ecosystems causing negative impacts on the ecological and economic values of sagebrush steppe of the western USA. Although our knowledge of the drivers of the regional distribution of B. tectorum has improved, we have yet to determine the relative importance of climate and local factors causing B. tectorum abundance and impact. To address this, we sampled 555 sites distributed geographically and ecologically throughout the sagebrush steppe. We recorded the canopy cover of B. tectorum, as well as local substrate and vegetation characteristics. Boosted regression tree modeling revealed that climate strongly limits the transformative ability of B. tectorum to a portion of the sagebrush steppe with dry summers (that is, July precipitation <10 mm and the driest annual quarter associated with a mean temperature >15°C) and low native grass canopy cover. This portion includes the Bonneville, Columbia, Lahontan, and lower Snake River basins. These areas are likely to require extreme efforts to reverse B. tectorum transformation. Our predictions, using future climate conditions, suggest that the transformative ability of B. tectorum may not expand geographically and could remain within the same climatically suitable basins. We found B. tectorum in locally disturbed areas within or adjacent to all of our sample sites, but not necessarily within sagebrush steppe vegetation. Conversion of the sagebrush steppe by B. tectorum, therefore, is more likely to occur outside the confines of its current climatically optimal region because of site-specific disturbances, including invasive species control efforts and sagebrush steppe mismanagement, rather than climate change.     

Oxidative DNA damage in peripheral blood cells in type 2 diabetes mellitus: higher vulnerability of polymorphonuclear leukocytes

Since oxidative stress is thought to play an important role in the pathogenesis and complications of diabetes, we used the comet assay (single cell alkaline gel electrophoresis) to evaluate DNA strand breaks and DNA base oxidation, measured as FPG (formamidopyrimidine DNA glycosylase)-sensitive sites, in peripheral blood cells (PBC) from type 2 diabetes patients and healthy controls. Oxidative DNA damage in leukocytes was increased in diabetic compared to normal subjects. However, no differences in the levels of DNA damage in isolated lymphocytes were found between the two groups. These data indicate a higher vulnerability to oxidative damage of polymorphonuclear as compared to mononuclear leukocytes in type 2 diabetes. Thus, the measurement of oxidative DNA damage in leukocytes by means of the comet assay is a suitable marker for the evaluation of systemic oxidative stress in diabetic patients.