The role of the yeast cleavage and polyadenylation factor subunit Ydh1p/Cft2p in pre-mRNA 3' end formation

Cleavage and polyadenylation factor (CPF) is a multi‐protein complex that functions in pre‐mRNA 3′‐end formation and in the RNA polymerase II (RNAP II) transcription cycle. Ydh1p/Cft2p is an essential component of CPF but its precise role in 3′‐end processing remained unclear. We found that mutations in YDH1 inhibited both the cleavage and the polyadenylation steps of the 3′‐end formation reaction in vitro. Recently, we demonstrated that an important function of CPF lies in the recognition of poly(A) site sequences and RNA binding analyses suggesting that Ydh1p/Cft2p interacts with the poly(A) site region. Here we show that mutant ydh1 strains are deficient in the recognition of the ACT1 cleavage site in vivo. The C‐terminal domain (CTD) of RNAP II plays a major role in coupling 3′‐end processing and transcription. We provide evidence that Ydh1p/Cft2p interacts with the CTD of RNAP II, several other subunits of CPF and with Pcf11p, a component of CF IA. We propose that Ydh1p/Cft2p contributes to the formation of important interaction surfaces that mediate the dynamic association of CPF with RNAP II, the recognition of poly(A) site sequences and the assembly of the polyadenylation machinery on the RNA substrate.

Functions for S. cerevisiae Swd2p in 3' end formation of specific mRNAs and snoRNAs and global histone 3 lysine 4 methylation

The Saccharomyces cerevisiae WD-40 repeat protein Swd2p associates with two functionally distinct multiprotein complexes: the cleavage and polyadenylation factor (CPF) that is involved in pre-mRNA and snoRNA 3′ end formation and the SET1 complex (SET1C) that methylates histone 3 lysine 4. Based on bioinformatic analysis we predict a seven-bladed β-propeller structure for Swd2p proteins. Northern, transcriptional run-on and in vitro 3′ end cleavage analyses suggest that temperature sensitive swd2 strains were defective in 3′ end formation of specific mRNAs and snoRNAs. Protein–protein interaction studies support a role for Swd2p in the assembly of 3′ end formation complexes. Furthermore, histone 3 lysine 4 di-and tri-methylation were adversely affected and telomeres were shortened in swd2 mutants. Underaccumulation of the Set1p methyltransferase accounts for the observed loss of SET1C activity and suggests a requirement for Swd2p for the stability or assembly of this complex. We also provide evidence that the roles of Swd2p as component of CPF and SET1C are functionally independent. Taken together, our results establish a dual requirement for Swd2p in 3′ end formation and histone tail modification.

Echium oil provides no benefit over linseed oil for (n-3) long-chain PUFA biosynthesis in rainbow trout

The implementation of alternative lipid sources for use in aquaculture is of considerable interest globally. However, the possible benefit of using stearidonic acid (SDA)–rich fish oil (FO) alternatives has led to scientific confusion. Two hundred and forty rainbow trout (Oncorhynchus mykiss) were fed 1 of 4 diets (3 replicate tanks/treatment) containing either FO, linseed oil (LO), echium oil, or mixed vegetable oil (72% LO, 23% sunflower oil, and 6% canola oil) as the dietary lipid source (16.5%) for 73 d to investigate the competition and long-chain PUFA (LC-PUFA) biosynthesis between the fatty acid substrates α-linolenic acid (ALA) and SDA. SDA was more efficiently bioconverted to LC-PUFA compared with ALA. However, when the dietary lipid sources were directly compared, the increased provision of C18 PUFA within the LO diet resulted in no significant differences in (n-3) LC-PUFA content compared with fish fed the other diets. This study therefore shows that, rather than the previously speculated substrate competition, the limiting process in the apparent in vivo (n-3) LC-PUFA biosynthesis appears to be substrate availability. Rainbow trout fed the SDA- and ALA-rich dietary lipid sources subsequently had similar significant reductions in (n-3) LC-PUFA compared with fish fed the FO diet, therefore providing no additional dietary benefit on (n-3) LC-PUFA concentrations.

Concomitant reactivity of the m-Terphenylindium dihydroxide [2,6-Mes2C6H3In(OH)2]4 toward carbon dioxide and ethylene glycol

The stepwise reaction of [2,6-Mes₂C₆H₃In(OH)₂]₄ with carbon dioxide and ethylene glycol proceeded with the formation of (2,6-Mes₂C₆H₃In)₄(CO₃)₂(OH)₄(H₂O)₂ (1) and (2,6-Mes₂C₆H₃In)₄(OCH₂CH₂O)₂(OH)₄ (2), respectively, and eventually produced (2,6-Mes₂C₆H₃In)₄(CO₃)₂(OCH₂CH₂OH)₂(OH)₂ (3). Attempts to liberate ethylene carbonate upon heating of 3 were unsuccessful.

The electrochemical surface forces apparatus : the effect of surface roughness, electrostatic surface potentials, and anodic oxide growth on interaction forces, and friction between dissimilar surfaces in aqueous solutions

We present a newly designed electrochemical surface forces apparatus (EC-SFA) that allows control and measurement of surface potentials and interfacial electrochemical reactions with simultaneous measurement of normal interaction forces (with nN resolution), friction forces (with μN resolution), and distances (with Å resolution) between apposing surfaces. We describe three applications of the developed EC-SFA and discuss the wide-range of potential other applications. In particular, we describe measurements of (1) force–distance profiles between smooth and rough gold surfaces and apposing self-assembled monolayer-covered smooth mica surfaces; (2) the effective changing thickness of anodically growing oxide layers with Å-accuracy on rough and smooth surfaces; and (3) friction forces evolving at a metal–ceramic contact, all as a function of the applied electrochemical potential. Interaction forces between atomically smooth surfaces are well-described using DLVO theory and the Hogg–Healy–Fuerstenau approximation for electric double layer interactions between dissimilar surfaces, which unintuitively predicts the possibility of attractive double layer forces between dissimilar surfaces whose surface potentials have similar sign, and repulsive forces between surfaces whose surface potentials have opposite sign. Surface roughness of the gold electrodes leads to an additional exponentially repulsive force in the force–distance profiles that is qualitatively well described by an extended DLVO model that includes repulsive hydration and steric forces. Comparing the measured thickness of the anodic gold oxide layer and the charge consumed for generating this layer allowed the identification of its chemical structure as a hydrated Au(OH)3 phase formed at the gold surface at high positive potentials. The EC-SFA allows, for the first time, one to look at complex long-term transient effects of dynamic processes (e.g., relaxation times), which are also reflected in friction forces while tuning electrochemical surface potentials.

Leukocyte numbers and function in subjects eating n-3 enriched foods: selective depression of natural killer cell levels

Introduction : While consumption of omega-3 long-chain polyunsaturated fatty acids (n-3 LCPUFA) has been recommended for those at risk of inflammatory disease such as rheumatoid arthritis, the mechanism of their anti-inflammatory effect remains to be clearly defined, particularly in relation to the dose and type of n-3 LCPUFA. The objective of this study was to determine whether varying the levels of n-3 LCPUFA in erythrocyte membrane lipids, following dietary supplementation, is associated with altered numbers and function of circulating leukocytes conducive to protection against inflammation. Methods : In a double-blind and placebo-controlled study, 44 healthy subjects aged 23 to 63 years consumed either standard or n-3 LCPUFA-enriched versions of typical processed foods, the latter allowing a target daily consumption of 1 gram n-3 LCPUFA. After six months, peripheral blood leukocyte and subpopulation proportions and numbers were assessed by flow cytometry. Leukocytes were also examined for lymphoproliferation and cytokine production, neutrophil chemotaxis, chemokinesis, bactericidal, adherence and iodination activity. Erythrocytes were analyzed for fatty-acid content. Results : Erythrocyte n-3 LCPUFA levels were higher and absolute leukocyte and lymphocyte numbers were lower in subjects consuming n-3 enriched foods than in controls. There were no changes in the number of neutrophils, monocytes, T cells (CD3+), T-cell subsets (CD4+, CD8+) and B cells (CD19+). However, natural killer (NK) (CD3-CD16+CD56+) cell numbers were lower in n-3 supplemented subjects than in controls and were inversely related to the amount of eicosapentaenoic acid or docosahexaenoic acid in erythrocytes. No significant correlations were found with respect to lymphocyte lymphoproliferation and production of IFN-γ and IL-2, but lymphotoxin production was higher with greater n-3 LCPUFA membrane content. Similarly, neutrophil chemotaxis, chemokinesis, bactericidal activity and adherence did not vary with changes in erythrocyte n-3 LCPUFA levels, but the iodination reaction was reduced with higher n-3 LCPUFA content. Conclusion : The data show that regular long-term consumption of n-3 enriched foods leads to lower numbers of NK cells and neutrophil iodination activity but higher lymphotoxin production by lymphocytes. These changes are consistent with decreased inflammatory reaction and tissue damage seen in patients with inflammatory disorders receiving n-3 LCPUFA supplementation.

Gay men's estimates of the likelihood of HIV transmission in sexual behaviours

In 3 studies we recorded gay men's estimates of the likelihood that HIV would be transmitted in various sexual behaviours. In Study 1 (data collected 1993, n=92), the men were found to believe that transmissibility is very much greater than it actually is; that insertive unprotected anal intercourse (UAI) by an HIV-infected partner is made safer by withdrawal before ejaculation, and very much safer by withdrawal before either ejaculation or pre-ejaculation; that UAI is very much safer when an infected partner is receptive rather than insertive; that insertive oral sex by an infected partner is much less risky than even the safest variant of UAI; that HIV is less transmissible very early after infection than later on; and that risk accumulates over repeated acts of UAI less than it actually does. In Study 2 (data collected 1997/8, n=200), it was found that younger and older uninfected men generally gave similar estimates of transmissibility, but that infected men gave somewhat lower estimates than uninfected men; and that estimates were unaffected by asking the men to imagine that they themselves, rather than a hypothetical other gay man, were engaging in the behaviours. Comparison of the 1993 and 1997/8 results suggested that there had been some effect of an educational campaign warning of the dangers of withdrawal; however, there had been no effect either of a campaign warning of the dangers of receptive UAI by an infected partner, or of publicity given to the greater transmissibility of HIV shortly after infection. In Study 3 (data collected 1999, n=59), men induced into a positive mood were found to give lower estimates of transmissibility than either men induced into a neutral mood or men induced into a negative mood. It is argued that the results reveal the important contribution made to gay men's transmissibility estimates by cognitive strategies (such as the 'availability heuristic' and 'anchoring and adjustment') known to be general characteristics of human information-processing. Implications of the findings for AIDS education are discussed.

Culture-specific variation in the flavor profile of soymilks

A modified quantitative descriptive analysis (QDA) method was used to determine sensory profiles of 8 soymilk products: 3 manufactured in Australia, 3 manufactured in Singapore, 1 manufactured in Malaysia, and 1 manufactured in Hong Kong. A panel (n= 7) was selected, trained in descriptive profiling of soymilk, and developed a soymilk language that was used to evaluate the flavor attributes of the soymilk products. A repeated-measure ANOVA showed highly reproducible panel performance, and significant differences in soymilk attributes among all soymilks. A principal component analysis (PCA) revealed 2 main groupings among the soymilks that corresponded to cultural origin: Australia and Asia (Singapore and Hong Kong/Malaysia). Products from Australia were significantly stronger in milky, astringent, salty notes and pale in color, while products from Asia were significantly stronger in beany, cooked beans, sweet, and pandan notes (P < 0.05). In addition, the Asian soymilks could be separated into 2 subgroups, with Singaporean soymilks having deeper color, greater viscosity, and less green flavor than Hong Kong/Malaysia soymilks. Australian produced soymilk is bovine-milk-like compared with Asian soymilk, presumably due to bovine milk being the primary source of milk in Australia. We conclude that culture-specific flavor preferences are a determining factor in flavor profiles of soymilks from geographically distinct regions.

Active transport to work in Australia: is it all downhill from here?

Physical inactivity is increasing in Australia and active forms of transportation may be one way to increase the working population’s daily physical activity. We used travel-to-work data from employed persons aged 15 years and over participating in the 1996 (n=7,636,319) and 2001(n=8,298,606) Australian censuses to determine prevalence and trends in walking and cycling to work by state and gender, and differences in prevalence by age. In 2001, 3.8% of Australians walked to work and <1% cycled. Over 64% travelled to work by car. There have been small declines in walking (men and women) and cycling (men) over the 5-years from 1996 to 2001. People were more likely to walk or cycle to work if they lived in the Northern Territory, if they were male or if they were aged 15 to 24 years. They were more likely to travel by car if they lived in the Australian Capital Territory, if they were male, or if they were aged 45-54 years. Few people walk or cycle to work in Australia. Efforts to encourage active transportation are urgently needed.

Acute exercise and GLUT4 expression in human skeletal muscle: influence of exercise intensity

To examine the influence of exercise intensity on the increases in vastus lateralis GLUT4 mRNA and protein after exercise, six untrained men exercised for 60 min at 39 ± 3% peak oxygen consumption (VO2 peak) (Lo) or 27 ± 2 min at 83 ± 2% VO2 peak (Hi) in counterbalanced order. Preexercise muscle glycogen levels were not different between trials (Lo: 408 ± 35 mmol/kg dry mass; Hi: 420 ± 43 mmol/kg dry mass); however, postexercise levels were lower (P < 0.05) in Hi (169 ± 18 mmol/kg dry mass) compared with Lo (262 ± 35 mmol/kg dry mass). Thus calculated muscle glycogen utilization was greater (P < 0.05) in Hi (251 ± 24 mmol/kg) than in Lo (146 ± 34). Exercise resulted in similar increases in GLUT4 gene expression in both trials. GLUT4 mRNA was increased immediately at the end of exercise (~2-fold; P < 0.05) and remained elevated after 3 h of postexercise recovery. When measured 3 h after exercise, total crude membrane GLUT4 protein levels were 106% higher in Lo (3.3 ± 0.7 vs. 1.6 ± 0.3 arbitrary units) and 61% higher in Hi (2.9 ± 0.5 vs. 1.8 ± 0.5 arbitrary units) relative to preexercise levels. A main effect for exercise was observed, with no significant differences between trials. In conclusion, exercise at ~40 and ~80% VO2 peak, with total work equal, increased GLUT4 mRNA and GLUT4 protein in human skeletal muscle to a similar extent, despite differences in exercise intensity and duration.

Extremely high conservation in the untranslated Region as well as the coding region of CNP mRNAs throughout elasmobranch species

C-type natriuretic peptide (CNP) is a crucial osmoregulatory hormone in elasmobranchs, participating in salt secretion and drinking. In contrast to teleosts and tetrapods in which the NP family is composed of a group of structurally related peptides, we have shown that CNP is the sole NP in sharks. In the present study, CNP cDNAs were cloned from four species of batoids, another group of elasmobranchs. The cloned batoid CNP precursors contained a plausible mature peptide of 22 amino acid residues that is identical to most shark CNP-22s, but five successive amino acids were consistently deleted in the prosegment compared with shark precursors, supporting the diphyletic classification of sharks and rays. In addition, molecular phylogenetic trees of CNP precursors were consistent with a diphyletic interpretation. Except for the deletion, the nucleotide and deduced amino acid sequences of the CNP cDNAs are extremely well-conserved among all elasmobranch species, even between sharks and rays. Surprisingly, high conservation is evident not only for the coding region, but also for the untranslated regions. It is most likely that the high conservation is due to the low nucleotide substitution rate in the elasmobranch genome, and high selection pressure. The 3′-untranslated region of the elasmobranch CNP cDNAs contained three to six repeats of the ATTTA motif that is associated with the regulation of mRNA stability and translation efficiency. Alternative polyadenylation sites were also found; the long 3′-untranslated region contains a core of ATTTA motifs while the short form has only one or no ATTTA motif, indicating that the post-transcriptional modification of mRNA is important for regulation of CNP synthesis. These characteristics in the 3′-untranslated region were conserved among all elasmobranch CNP cDNAs. Since CNP has been implicated as a fast-acting hormone to facilitate salt secretion from the rectal gland, the conserved 3′-untranslated region most likely contributes to rapid regulation of CNP synthesis in elasmobranchs in response to acute changes in internal and external environments.

Insulin signaling after exercise in insulin receptor substrate-2-deficient mice

The period immediately after exercise is characterized by enhanced insulin action in skeletal muscle, and on the molecular level, by a marked increase in insulin-stimulated, phosphotyrosine-associated phosphatidylinositol (PI) 3-kinase activity. Because the increase in PI 3-kinase activity cannot be explained by increased insulin receptor substrate (IRS)-1 signaling, the present study examined whether this effect is mediated by enhanced IRS-2 signaling. In wild-type (WT) mice, insulin increased IRS-2 tyrosine phosphorylation (2.5-fold) and IRS-2-associated PI 3-kinase activity (3-fold). Treadmill exercise, per se, had no effect on IRS-2 signaling, but in the period immediately after exercise, there was a further increase in insulin-stimulated IRS-2 tyrosine phosphorylation (3.5-fold) and IRS-2-associated PI 3-kinase activity (5-fold). In IRS-2-deficient (IRS-2-/-) mice, the increase in insulin-stimulated, phosphotyrosine-associated PI 3-kinase activity was attenuated as compared with WT mice. However, in IRS-2-/- mice, the insulin-stimulated, phosphotyrosine-associated PI 3-kinase response after exercise was slightly higher than the insulin-stimulated response alone. In conclusion, IRS-2 tyrosine phosphorylation and associated PI 3-kinase activity are markedly enhanced by insulin in the immediate period after exercise. IRS-2 signaling can partially account for the increase in insulin-stimulated phosphotyrosine-associated PI 3-kinase activity after exercise.

Differences in how english and german speakers talk and reason about CAUSE

This research identifies how English and German speakers differ in the range of situations they describe as causal and how these difference may influence causal reasoning. In Experiments 1 and 2, English and German speakers described 3D animations of complex events using CAUSE verbs (cause, get) and ENABLE verbs (let, enable). As predicted, English speakers used CAUSE verbs to describe a wider range of events than German speakers. In Experiment 3, English and German speakers viewed 3D animations of CAUSE and ENABLE events and then estimated the likelihood of the effect (E) in the presence of the affector (A), p(E|A), in hypothetical situations similar to the one they just saw. Given the results of Experiments 1 and 2, we predicted that German speakers’ estimates of p(E|A) would be higher than English speakers’ estimates of p(E|A) for ENABLE events, but not necessarily for CAUSE events. The results were as predicted. The findings suggest that English and German speakers differ in the range of situations they describe as causal and that these differences in linguistic coding may lead to differences in causal reasoning.

Biology curriculum for the senior secondary level in Victoria, Australia

In Victoria, the Victorian Certificate of Education(VCE) is most common among certificates required to apply any tertiary institute in the Victoria State. Thus, the number of students who take the VCE course is larger than other courses in senior secondary schools. VCE Biology is one of the subjects in natural science area. The subject consists of 4 units: Unit 1 is ecology oriented, Unit 2 is cell biology oriented, Unit 3 is physiology and developmental biology oriented, and Unit 4 is systematics, genetics and evolution oriented. One of the distinctive features of the VCE Biology is its assignment. Three or four tasks are prepared in each unit of the subject. In order to complete the assignment, students should carry out some laboratory work, field studies and investigations to collect data and information from a number of sources. They also need to analyze data to write some reports. In Unit 3 and 4, Common Assessment Tasks(CATs), which include writing report and paper test, and prepared. Another distinctive feature of the curriculum is that there are some applied biological aspects in the contents of each unit.

Pregnancy and lactation have no long-term deleterious effect on measures of bone mineral in healthy women: a twin study

BACKGROUND: The long-term effects of pregnancy and lactation on measures of bone mineral in women remain unclear.

OBJECTIVE: We studied whether pregnancy or lactation has deleterious long-term effects on bone mineral in healthy women.

DESIGN: We measured bone mineral density (BMD; g/cm(2)) in women aged > or = 18 y. Analyses were performed on 3 data sets: study 1, 83 female twin pairs (21 monozygous and 62 dizygous) aged (x +/- SD) 42.2 +/- 15.5 y who were discordant for ever having been pregnant beyond 20 wk; study 2, 498 twin pairs aged 42.3 +/- 15.0 y; and study 3, 1354 individual twins, their siblings, and family members.

RESULTS: In study 1, there were no significant within-pair differences in unadjusted BMD or BMD adjusted for age, height, and fat mass at the lumbar spine or total-hip or in total-body bone mineral content (BMC; kg) (paired t tests). In study 2, there was no significant within-pair difference in measures of bone mineral or body composition related to the within-pair difference in number of pregnancies. In study 3, subjects with 1 or 2 (n = 455) and > or = 3 pregnancies (n = 473) had higher adjusted lumbar spine BMD (2.9% and 3.8%, respectively; P = 0.001) and total-body BMC (2.2% and 3.1%; P < 0.001) than did nulliparous women (n = 426). Parous women who breast-fed had higher adjusted total-body BMC (2.6%; P = 0.005), total-hip BMD (3.2%; P = 0.04), and lower fat mass (10.9%; P = 0.01) than did parous non-breast-feeders.

CONCLUSION: We found no long-term detrimental effect of pregnancy or breast-feeding on bone mineral measures.