Might the kinetic behavior of hormone-sensitive lipase reflect the absence of the lid domain?

Hormone-sensitive lipase (HSL) is thought to contribute importantly to the mobilization of fatty acids from the triacylglycerols (TAGs) stored in adipocytes, providing the main source of energy in mammals. To investigate the HSL substrate specificity more closely, we systematically assessed the lipolytic activity of recombinant human HSL on solutions and emulsions of various vinyl esters and TAG substrates, using the pH-stat assay technique. Recombinant human HSL activity on solutions of partly soluble vinyl esters or TAG was found to range from 35 to 90% of the maximum activity measured with the same substrates in the emulsified state. The possible existence of a lipid-water interface due to the formation of small aggregates of vinyl esters or TAG in solution may account for the HSL activity observed below the solubility limit of the substrate. Recombinant human HSL also hydrolyzes insoluble medium- and long-chain acylglycerols such as trioctanoylglycerol, dioleoylglycerol, and olive oil, and can therefore be classified as a true lipase. Preincubation of the recombinant HSL with a serine esterase inhibitor such as diethyl p-nitrophenyl phosphate in 1:100 molar excess leads to complete HSL inhibition within 15 min. This result indicates that the catalytic serine of HSL is highly reactive and that it is readily accessible. Similar behavior was also observed with lipases with no lid domain covering their active site, or with a deletion in the lid domain. The 3-D structure of HSL, which still remains to be determined, may therefore lack the lid domain known to exist in various other lipases.     

How to assess the prediction accuracy of species presence–absence models without absence data?

It is very common that only presence data are available in ecological niche modeling. However, most existing methods for evaluating the accuracy of presence–absence (binary) predictions of species require presence–absence data. The aim of this study is to present a new method for accuracy assessment that does not rely on absence data. Two new statistics F_pb and F_cpb were derived based on presence–background data. With generated six virtual species, we used DOMAIN, generalized linear modeling (GLM), and maximum entropy (MAXENT) to produce different species presence–absence predictions. To investigate the effectiveness of the new statistics in accuracy assessment, we used F_pb, F_cpb, the traditional F‐measure (F), kappa coefficient, true skill statistic (TSS), area under the receiver operating characteristic curve (AUC), and the contrast validation index (CVI) to evaluate the accuracy of predictions, and the behaviors of these accuracy measures were compared. The effectiveness of F_pb for threshold selection and estimation of species prevalence was also investigated. Experimental results show that F_cpb is an estimate of F. The Pearson's correlation coefficient (COR) between F_cpb and F is 0.9882, with a root‐mean‐square error (RMSE) of 0.0171. In general, F_pb, F_cpb, F, kappa coefficient, TSS, and CVI can sort models by the accuracy of binary prediction, but AUC is not appropriate to evaluate the accuracy of binary prediction. For DOMAIN, GLM, and MAXENT, finding the threshold by maximizing F_pb and by maximizing F result in similar accuracies. In addition, the estimation of species prevalence based on binary output with maximizing F_pb as the thresholding method is significantly more accurate than simply averaging the original continuous output. The best estimate of prevalence is provided by the binary output of MAXENT, with an RMSE of 0.0116. Finally, we conclude that the new method is promising in accuracy assessment, threshold selection, and estimation of species prevalence, all of which are important but challenging problems with presence‐only data. Because it does not require absence data, the new method will have important applications in ecological niche modeling.     

Laboratory rat selection for the trait “the absence of audiogenic seizure proneness”

The hybrids between Krushinsky-Molodkina (KM) inbred strain, selected for high predisposition to audiogenic epilepsy (AE), and Wistar rats non-prone to audiogenic seizure were the initial population for selection. Rats were selected for the trait ??the absence of audiogenic seizure proneness??. The creation of such strain in which the significant proportion of animals develop no AE in response to sound and share partly the genetic background of the KM strain is very important for the correct use of KM strain as the laboratory model of seizure states. As alleles which determine the AE proneness are recessive the selection for the ??opposite?? trait proceeds necessarily slow.     

Stability of recombinant bovine interferon-γ antiviral activity in the absence of stabilizing additives

The stability of recombinant bovine interferon-γ (rbIFN-γ) produced by a baculovirus expression system was investigated under different storage conditions: freezing-thawing and storage for 30 days at temperatures of -80, 4, 25, and 37°C. Antiviral activity was not significantly decreased by freeze-thawing at least five times. Furthermore, although not statistically different, antiviral activity gradually decreased as temperature increased. These findings suggest that rbIFN-γ possesses high thermal and freeze-thaw stability.     

Do Svalbard reindeer regulate standing crop in the absence of predators?

The Svalbard reindeer is the only mammalian herbivore in Adventdalen (78°N), Svalbard, where it has no natural predators. To test if herbivores in the absence of predators regulate standing crop to a level independent of productivity, which is one of the predictions of the “exploitation ecosystems” model, herbivore exclosures were set up in 1992 in Salix heath, Luzula heath, Cassiope heath, and Alopecurus meadow in Adventdalen. Standing crop of vascular plants was harvested and measured inside and outside the exclosures in 1994, when the reindeer population was at peak density (ca 5.4 animals km⁻²), and in 1996, when the reindeer density was about 30% lower (ca 3.7 animals km⁻²). Standing dead material was reduced by grazing in the Luzula heath in 1994. However, we found no effect of grazing, year, or interactions between grazing and year on live standing crop. Also contrary to the predictions from the model, differences in standing crop between vegetation types were highly significant. Mean biomass of plant material was lowest in the Alopecurus meadow (36 g m⁻²), two fold higher in the Luzula heath, and about threefold higher in the Salix heath and Cassiope heath, indicating that reindeer do not regulate standing crop to the same level on a local scale. The predictive power of the “exploitation ecosystems” model is low due to lack of recognition of the importance of plant chemistry, plant compensation ability, variation in forage availability during the year, parasites functioning as predators, and adverse weather conditions, which may cause density-independent variations in fecundity and mortality of reindeer. Received: 28 December 1997 / Accepted: 6 April 1998     

E. coli DNA replication in the absence of free β clamps

During DNA replication, repetitive synthesis of discrete Okazaki fragments requires mechanisms that guarantee DNA polymerase, clamp, and primase proteins are present for every cycle. In Escherichia coli, this process proceeds through transfer of the lagging-strand polymerase from the β sliding clamp left at a completed Okazaki fragment to a clamp assembled on a new RNA primer. These lagging-strand clamps are thought to be bound by the replisome from solution and loaded a new for every fragment. Here, we discuss a surprising, alternative lagging-strand synthesis mechanism: efficient replication in the absence of any clamps other than those assembled with the replisome. Using single-molecule experiments, we show that replication complexes pre-assembled on DNA support synthesis of multiple Okazaki fragments in the absence of excess β clamps. The processivity of these replisomes, but not the number of synthesized Okazaki fragments, is dependent on the frequency of RNA-primer synthesis. These results broaden our understanding of lagging-strand synthesis and emphasize the stability of the replisome to continue synthesis without new clamps.     

Anti-Semitism in Britain: continuity and the absence of a resurgence?

Abstract

It has become the orthodoxy in recent years to assume that anti-Semitism globally is not only rising but also taking a new form – it is a ‘new anti-Semitism’ or even a new phenomenon: Judeophobia. This article takes a different perspective. It initially covers approaches to anti-Semitism and how, especially in the light of the Holocaust, it has been viewed academically as no longer the fault of the Jews but as a natural and constant feature of history since antiquity. A critique is provided of the idea of a continuous history of anti-Semitism and of the metaphors used to describe it. There then follows a case study of anti-Semitism in Britain. The British case is valuable as it is seen as a key example of the ‘new anti-Semitism’, and one that is more striking given the alleged absence of previous hostility towards Jews in that country. By employing a comparative approach – both temporal and in relation to responses to other groups – change and continuity are charted through a study of racial violence. Such comparisons, it is argued, allow a more nuanced and balanced analysis of this issue, which has created much alarm and little sober reflection.     

Does the speciation clock tick more slowly in the absence of heteromorphic sex chromosomes?

Squamates may be an attractive group in which to study the influence of sex chromosomes on speciation rates because of the repeated evolution of heterogamety (both XY and ZW), as well as an apparently large number of taxa with environmental sex-determination.     

The absence of the CpNpG methylation at the 5′-terminal region of the human calcitonin gene in norm and leukemias

The inner cytosine methylation was analyzed in the CCWGG sequences of the 5′-terminal region of the human calcitonin gene from peripheral blood and bone marrow cells in various forms of leukemia. Since these sequences remain nonmethylated both in the norm and in various leukemia forms, the CpG dinucleotide hypermethylation of the 5′-end of the human calcitonin gene, characteristic for the development of leukemias, does not spread to adjacent CpNpG sequences.     

Characterization of chicken Mda5 activity: regulation of IFN-β in the absence of RIG-I functionality

In mammals, Mda5 and RIG-I are members of the evolutionary conserved RIG-like helicase family that play critical roles in the outcome of RNA virus infections. Resolving influenza infection in mammals has been shown to require RIG-I; however, the apparent absence of a RIG-I homolog in chickens raises intriguing questions regarding how this species deals with influenza virus infection. Although chickens are able to resolve certain strains of influenza, they are highly susceptible to others, such as highly pathogenic avian influenza H5N1. Understanding RIG-like helicases in the chicken is of critical importance, especially for developing new therapeutics that may use these systems. With this in mind, we investigated the RIG-like helicase Mda5 in the chicken. We have identified a chicken Mda5 homolog (ChMda5) and assessed its functional activities that relate to antiviral responses. Like mammalian Mda5, ChMda5 expression is upregulated in response to dsRNA stimulation and following IFN activation of cells. Furthermore, RNA interference-mediated knockdown of ChMda5 showed that ChMda5 plays an important role in the IFN response of chicken cells to dsRNA. Intriguingly, although ChMda5 levels are highly upregulated during influenza infection, knockdown of ChMda5 expression does not appear to impact influenza proliferation. Collectively, although Mda5 is functionally active in the chicken, the absence of an apparent RIG-I-like function may contribute to the chicken's susceptibility to highly pathogenic influenza.     

Functional properties of the Cav1.2 calcium channel activated by calmodulin in the absence of α2δ subunits

Voltage-activated Ca_v1.2 calcium channels require association of the pore-forming α_1C subunit with accessory Ca_vβ and α₂δ subunits. Binding of a single calmodulin (CaM) to α_1C supports Ca²⁺-dependent inactivation (CDI). The human Ca_v1.2 channel is silent in the absence of Ca_vβ and/or α₂δ. Recently, we found that coexpression of exogenous CaM (CaM_ex) supports plasma membrane targeting, gating facilitation and CDI of the channel in the absence of Ca_vβ. Here we discovered that CaM_ex and its Ca²⁺-insensitive mutant (CaM₁₂₃₄) rendered active α_1C/Ca_vβ channel in the absence of α₂δ. Coexpression of CaM_ex with α_1C and β_2d in calcium-channel-free COS-1 cells recovered gating of the channel and supported CDI. Voltage-dependence of activation was shifted by ≈ +40 mV to depolarization potentials. The calcium current reached maximum at +40 mV (20 mM Ca²⁺) and exhibited approximately 3 times slower activation and 5 times slower inactivation kinetics compared to the wild-type channel. Furthermore, both CaM_ex and CaM₁₂₃₄ accelerated recovery from inactivation and induced facilitation of the calcium current by strong depolarization prepulse, the properties absent from the human vascular/neuronal Ca_v1.2 channel. The data suggest a previously unknown action of CaM that in the presence of Ca_vβ translates into activation of the α₂δ-deficient calcium channel and alteration of its properties.     

Secretion of gonadotrophins change during the luteal phase of the bovine oestrous cycle in the absence of corresponding changes in progesterone or 17β-oestradiol

The hypothesis in the present study was that changes in circulating luteinizing hormone (LH) and follicle stimulating hormone (FSH) would occur during the luteal phase of the oestrous cycle (Days 4–19; Day 0, day of behaviourial oestrus) that were not related to corresponding changes in concentrations of progesterone and 17β-oestradiol. The stage of the oestrous cycle of cows (n = 18) was synchronised to obtain cows that were on alternate days of the cycle. Blood samples were collected every other day at 15 min intervals for 12 h from all cows: Days 4, 6, 8, 10, 12, 14, 16, 18 (n = 9) and Days 5, 7, 9, 11, 13, 15, 17, 19 (n = 9). Concentrations of LH, FSH, 17β-oestradiol and progesterone were determined in these samples. Data were compared across days to determine when significant changes occurred in concentrations or patterns of secretion of the gonadotrophins and ovarian steroid hormones during the oestrous cycle. There were significant changes in mean concentrations of FSH in circulation between Days 6 and 12. The most striking changes in secretion of gonadotrophins that could not be explained by changes in gonadal steroids were the fluctuations in amplitude of LH pulses between Days 7 and 12. Amplitude of LH pulses increased between Days 7 and 11 and subsequently decreased between Days 11 and 12 of the oestrous cycle. Some changes in gonadotrophin secretion that occurred in the present study can be explained by fluctuations in concentrations of progesterone and 17β-oestradiol in circulation. Other changes cannot be explained by fluctuations in circulating concentrations of these steroids. We accept our hypothesis because the concomitant changes in mean concentration of FSH between Days 6 and 11 and amplitude of LH pulses between Days 7 and 12 of the bovine oestrous cycle cannot be explained by changes in circulating concentrations of progesterone and 17β-oestradiol.     

Does fitness erode in the absence of selection? An experimental test with Tribolium

In the absence of natural selection, average fitness in the population is expected to decline due to the accumulation of deleterious mutations. Replicate populations of flour beetles (Tribolium confusum) were maintained for 22 generations in the virtual absence of selection (random mating, favorable environment, excess of food, and mortality of only 3%). Larva-to-adult survivorship rates were similar in the stock population and selection-free populations. In contrast, starvation resistance of adult beetles from selection-free populations was significantly reduced (by more than 2% per generation). When tested in the favorable environment, beetles in one selection-free population had significantly slower development and smaller sizes of females than beetles from the stock population. Since such changes in these fitness components are usually maladaptive, they indicate possible erosion of fitness under relaxed selection at the rate of 0.1-0.2% per generation. No fitness erosion was detectable in the second selection-free population.