Isolation and1H-NMR spectroscopic identification of the glucose-containing lipid-linked precursor oligosaccharide ofN-linked carbohydrate chains

The lipid-linked precursor ofN-type glycoprotein oligosaccharides was isolated from porcine thyroid microsomes after in cubation with UDP[³H] Glucose. The carbohydrate was released from dolichol pyrophosphate by mild acid hydrolysis, purified by gel filtration and characterized by 500-MHz¹H-NMR spectroscopy in combination with enzymatic degradation. The parent oligosaccharide was found to be Glc₃Man₉Glc-NAc₂. The three glucose residues are present in the linear sequence Glcα1-2Glα1-3 Glc, the latter being α(1-3)-linked to one of the mannose residues. In order to establish the branch location of the triglucosyl unit, the parent compound was digested with jack-bean α-mannosidase. The oligosaccharide product was purified by gel filtration, and identified by¹H-NMR as Glc₃Man₅GlcNAc₂ lacking the mannose residues A, D₂, B and D₃. Therefore, the structure of the precursor oligosaccharide is as follows: $$\begin{gathered} c b a D_1 C 4 \hfill \\ Glc\alpha 1 - 2Glc\alpha 1 - 3Glc\alpha 1 - 3Man\alpha 1 - 2Man\alpha 1 - 2Man\alpha 1 \hfill \\ 3 \swarrow 3 2 1 \hfill \\ Man\alpha 1 - 2Man\alpha 1 Man\beta 1 - 4GlcNAc\beta 1 - 4GlcNAc \hfill \\ D_{2 } A 3 6 \hfill \\ Man\alpha 1 \hfill \\ 6 \hfill \\ Man\alpha 1 - 2Man\alpha 1 \nwarrow 4 \hfill \\ D_3 B \hfill \\ \end{gathered} $$      

Reprocessing data to form QALYs.

OBJECTIVES--To determine whether reprocessing data from published sources into quality adjusted life years (QALYs), as recommended in The QALY Toolkit, is a useful method of helping purchasing authorities to determine the most cost effective pattern of care to buy for their populations. SETTING--United Kingdom. DESIGN--The method was tested for six elective surgical conditions; data from published studies were reprocessed into the Rosser index, to obtain values for change in quality of life. These were then used to form QALYs. A small validation exercise was carried out. MAIN OUTCOME MEASURES--QALYs formed from the Rosser index. RESULTS--Published data could not be found for three interventions (cataract surgery, inguinal hernia repair, varicose vein surgery). For the remainder (prostatectomy, hip replacement, and knee replacement) data were found which could be reprocessed to form QALYs, though it was often hard to compare data from different studies and many assumptions had to be made. CONCLUSION--The value of QALY results obtained by this method is questionable, given the large number of assumptions which had to be made. For many interventions published data are unlikely to be available.     

Isolation and characterization of a diuretic peptide from Acheta domesticus. Evidence for a family of insect diuretic peptides.

A diuretic peptide (Acheta-DP) has been isolated from extracts of whole heads of the house cricket, Acheta domesticus. The native peptide increases both cyclic AMP production and the rate of fluid secretion by isolated Malpighian tubules in vitro to an extent comparable with those responses obtained with supra-maximal amounts of crude extracts of corpora cardiaca. The primary structure of Acheta-DP was established as a 46-residue amidated peptide: TGAQSLSIVAPLDVLRQRLMNELNRRRMRELQGSRIQQNRQLLTSI-NH2. Acheta-DP has 41% sequence identity with a diuretic peptide isolated from Manduca sexta, providing direct evidence for the presence of a family of diuretic peptides in insects.     

Influence of body size on oxygen consumption during bicycling

Energy in bicycling is primarily expended to overcome air resistance, which is proportional to a cyclist's surface area (SA). Thus we hypothesized that large cyclists should have a lower O2 consumption normalized to body weight (VO2/BW) than small cyclists because of the former's lower SA/BW. We measured the VO2/BW of small (BW = 59.4 +/- 4.1 kg) and large (BW = 84.4 +/- 3.2 kg) cyclists while they bicycled on a flat road at 10, 15, and 20 mph. The large cyclists had a 22% lower VO2/BW than the small cyclists at all speeds. However, the SA/BW ratio of the large cyclists was only 11% lower than that of the small cyclists. We then photographically determined the frontal area (FA) of the cyclists in a racing posture, and found that the large cyclists had a 16% lower FA/BW ratio than the small cyclists. We conclude that large cyclists are at a distinct advantage, in terms of VO2/BW, while bicycling on level roads, and this advantage is principally due to their lower FA/BW ratio.     

Transactional Sex Risk across a Typology of Rural and Urban Female Sex Workers in Indonesia: A Mixed Methods Study

Context-specific typologies of female sex workers (FSWs) are essential for the design of HIV intervention programming. This study develops a novel FSW typology for the analysis of transactional sex risk in rural and urban settings in Indonesia. Mixed methods include a survey of rural and urban FSWs (n = 310), in-depth interviews (n = 11), key informant interviews (n = 5) and ethnographic assessments. Thematic analysis categorises FSWs into 5 distinct groups based on geographical location of their sex work settings, place of solicitation, and whether sex work is their primary occupation. Multiple regression analysis shows that the likelihood of consistent condom use was higher among urban venue-based FSWs for whom sex work is not the only source of income than for any of the other rural and urban FSW groups. This effect was explained by the significantly lower likelihood of consistent condom use by rural venue-based FSWs (adjusted OR: 0.34 95% CI 0.13–0.90, p = 0.029). The FSW typology and differences in organisational features and social dynamics are more closely related to the risk of unprotected transactional sex, than levels of condom awareness and availability. Interventions need context-specific strategies to reach the different FSWs identified by this study''s typology.     

Comparison of insect kinin analogs with cis-peptide bond motif 4-aminopyroglutamate identifies optimal stereochemistry for diuretic activity

The insect kinins are present in a wide variety of insects and function as potent diuretic peptides, though they are subject to rapid degradation by internal peptidases. Insect kinin analogs incorporating stereochemical variants of (2S,4S)-4-aminopyroglutamate (APy), a cis-peptide bond motif, demonstrate significant activity in a cricket diuretic assay. Insect kinin analogs containing (2R,4R)-APy, (2S,4R)-APy and (2S,4S)-APy are essentially equipotent on an insect diuretic assay, with EC(50) values of about 10(-7)M, whereas the (2R,4S)-APy analog is at least 10-fold more potent (EC(50) = 7 x 10(-9)M). Conformational studies in aqueous solution indicate that the (2R,4S)-APy analog is considerably more flexible than the other three variants, which may explain its greater potency. The work identifies the optimal stereochemistry for the APy scaffold with which to design biostable, peptidomimetic analogs with the potential to disrupt critical insect kinin-regulated processes in insects.     

Beta-amino acid analogs of an insect neuropeptide feature potent bioactivity and resistance to peptidase hydrolysis

Insect neuropeptides of the insect kinin class share a common C-terminal pentapeptide sequence F(1)X(1)(2)X(2)(3)W(4)G(5)-NH(2) (X(2)(3) = P, S, A) and regulate such critical physiological processes as water balance and digestive enzyme release. Analogs of the insect kinin class, in which the critical residues of F(1), P(3), and W(4) were replaced with beta(3)-amino acid or their beta(2)-homo-amino acid variants, have been synthesized by the solid phase peptide strategy. The resulting single- and double-replacement analogs were evaluated in an insect diuretic assay and enzyme digestion trials. Analogs modified in the core P(3) position produce a potent and efficacious diuretic response that is not significantly different from that obtained with the endogenous achetakinin peptides. The analogs also demonstrate enhanced resistance to hydrolysis by ACE and NEP, endopeptidases that inactivate the natural insect neuropeptides. This paper describes the first instance of beta-amino acids analogs of an arthropod peptide that demonstrate significant bioactivity and resistance to peptidase degradation.     

Effect of recovery interval on multiple-bout sprint cycling performance after acute creatine supplementation

The purpose of this study was to examine the effect of varying recovery intervals on multiple-bout, short-duration, high-intensity cycling efforts of adult men supplemented with creatine (Cr) or a placebo (Pl). Thirty subjects underwent 3 trials of a maximal cycling protocol (T(0), T(1), T(2)). T(0) included V(O)2 max testing and familiarization with the sprint cycling protocol. T(1) consisted of 8 15-second bouts of sprint cycling exercise. Subjects were randomly assigned to recovery interval groups (1 minute, 3 minutes, 6 minutes), and Cr or Pl groups (0.3 g x kg(-1) x d(-1)). Posttesting (T(2)) took place 7 days after T(1) and consisted of an identical protocol as during T(1). Changes in mean power (MP), peak power (PP), and fatigue index (FI) were compared between trials. MP was significantly increased in Cr 1-minute, Cr 3-minute, and Pl 6-minute groups (p < 0.05). Significant PP increases were demonstrated in Cr 1-minute and Pl 6-minute groups (p < 0.05), and FI significantly increased in Pl 1-minute group (p < 0.05). Results indicate that Cr supplementation is effective in improving recovery from repeated sprint cycling performances when the recovery interval is of a short (<6 minutes) duration.