Identification of anchor points for chemical modification of a small cysteine-rich protein by using a cysteine scan

Chemical modifications of proteins are increasingly important in the development of protein drugs with fine-tuned properties. Regioselective modification, such as the chemoselective alkylation of an unpaired cysteine residue, is a prerequisite for obtaining homogenous protein products. The introduction of an unpaired Cys into the Cys-rich protein, insulin, was investigated by using a Cys scan. This was challenging as the introduced Cys could interfere with insulin's three existing disulfide bonds. However, eight insulin precursors were expressed in Saccharomyces cerevisiae with good yields. Although extensive post-translational modifications of the unpaired Cys were observed, the majority could be removed by selective reduction. An example Cys(7) insulin analogue was modified with a PEGylated maleimide moiety. The new variant was active in in vitro and in vivo models. Our results show that even small Cys-rich proteins can be expressed with additional unpaired Cys in meaningful yields and further chemically modified, while maintaining their biological activity.     

Fish Oil Supplementation During Late Pregnancy Does Not Influence Plasma Lipids or Lipoprotein Levels in Young Adult Offspring

Nutritional influences on cardiovascular disease operate throughout life. Studies in both experimental animals and humans have suggested that changes in the peri- and early post-natal nutrition can affect the development of the various components of the metabolic syndrome in adult life. This has lead to the hypothesis that n-3 fatty acid supplementation in pregnancy may have a beneficial effect on lipid profile in the offspring. The aim of the present study was to investigate the effect of supplementation with n-3 fatty acids during the third trimester of pregnancy on lipids and lipoproteins in the 19-year-old offspring. The study was based on the follow-up of a randomized controlled trial from 1990 where 533 pregnant women were randomized to fish oil (n = 266), olive oil (n = 136) or no oil (n = 131). In 2009, the offspring were invited to a physical examination including blood sampling. A total of 243 of the offspring participated. Lipid values did not differ between the fish oil and olive oil groups. The relative adjusted difference (95% confidence intervals) in lipid concentrations was −3% (−11; 7) for LDL cholesterol, 3% (−3; 10) for HDL cholesterol, −1% (−6; 5) for total cholesterol,−4% (−16; 10) for TAG concentrations, 2%(−2; 7) for apolipoprotein A1, −1% (−9; 7) for apolipoprotein B and 3% (−7; 15) in relative abundance of small dense LDL. In conclusion, there was no effect of fish oil supplementation during the third trimester of pregnancy on offspring plasma lipids and lipoproteins in adolescence.     

国内外简讯14则

△据中国橡胶工业协会轮胎分会统计 ,其 47家轮胎生产企业会员单位 2 0 0 3年共生产轮胎 1 0 24 83662条 ,与上年相比增长 1 0 68%,其中子午线轮胎产量为 485 88744条 ,同比增长38 1 2 %,而全钢载重子午线轮胎产量为1 0 1 1 91 1 6条 ,同比增长 64 7%;实现销售收入42 0 7769万元 ,同比增长 2 3 94%;出口交货量为35 72 695 3套 ,出口交货值为 9945 93万元 ,同比分别增长 1 3 32 %和 2 4 37%;实现利润总额和利税总额分别为 1 5 80 78万和 40 65 79万元 ,同比分别增长 1 9 69%和 5 5 2 %。(中国橡胶工业协会轮胎分会　蔡为民供稿 )…     

Gustatory sweating recurring on peritoneal dialysis but resolving during periods of hemodialysis

Gustatory sweating is a rare disorder characterized by profuse sweating on the forehead, face, scalp, and neck occurring soon after ingesting food, which has been reported in diabetic patients. The mechanism is thought to be triggered by taste buds and not gastric stimulation. We report a case where gustatory sweating repeatedly developed on peritoneal dialysis that resolved on periods of hemodialysis. A 32-year-old woman with diabetic end-stage renal disease developed gustatory sweating shortly after beginning continuous ambulatory peritoneal dialysis despite excellent clearances. After 5 months, she changed to hemodialysis for 2 months and noticed complete resolution of her gustatory sweating; however, after her return to peritoneal dialysis 2 months later, her gustatory sweating recurred. While on peritoneal dialysis, she was treated with clonidine, which resulted in improvement but not resolution of her symptoms as had occurred on hemodialysis. Another period on hemodialysis resulted in the resolution of her symptoms that returned again after restarting peritoneal dialysis. Clonidine provided incomplete relief while topical glycopyrrolate was effective and without complications. We report recurrent gustatory sweating on peritoneal dialysis that resolved with hemodialysis. We have no data to suggest that intra-abdominal stimulation played a role, but rather that despite excellent clearances neuropathy may have played a role. Treatment with topical glycopyrrolate may be safe and effective given every third day if clonidine is ineffective.     

Timing of onset of somatotropin supplementation on reproductive performance in dairy cows

The objective of this field trial was to determine whether delaying the start of recombinant bovine somatotropin (rbST) supplementation from 9 to 10 wk postpartum to 17 to 18 wk postpartum would improve reproductive performance in lactating dairy cows. Cows from nine herds (n = 798 cows; 766 Holsteins, 32 Jerseys) were assigned at random to receive rbST supplementation at 14-d intervals beginning during wk 9 to 10 (n = 399) or wk 17 to 18 (n = 399) after calving. Effects of herd, season of calving, parity, and onset of rbST supplementation (9 to 10 wk vs. 17 to 18 wk) on days to first service and days open were determined. In primiparous but not multiparous cows, there tended to be fewer days to first service and fewer days open when onset of rbST supplementation was delayed. Percentages of cows pregnant at 150, 200, and 250 d postpartum were also examined. Time of onset of rbST did not affect percentages of multiparous cows pregnant at 150, 200, and 250 d postpartum. However, there appeared to be a slight tendency for percentages of pregnant primiparous cows to be greater at 200 and 250 d postpartum for those receiving rbST supplementation beginning at 17 to 18 wk compared to those receiving rbST starting at 9 to 10 wk. In conclusion, delaying the start of rbST supplementation to wk 17 to 18 postpartum had no beneficial effect on reproductive performance of multiparous cows but tended to improve some measures of reproductive performance in primiparous cows.     

Essential fatty acid deficiency induces fatty acid desaturase expression in rat epididymis, but not in testis

On the molecular level, essential fatty acid deficiency (EFAD) has been associated with induced fatty acid (FA) desaturase expression and activity in several tissues. However, there seem to be exceptions. In the present study, we examine the effects of EFAD in the male rat genital tract, combining FA analysis, gene expression studies, and morphological evaluation of epididymal spermatozoa. When feeding 21-day-old Wistar rats, a fat-free diet for 6 weeks, an increase in 18:1n-9 and 20:3n-9 and a concomitant decrease in the 18:2n-6 and 20:4n-6 species are seen in testis, as well as in liver. However, with regard to desaturase expression the rat testis seems to be unresponsive to EFAD conditions, in contrast to other organs studied. In the sexually mature testis none of the desaturases (SCD1, SCD2, D5D, or D6D) are induced in response to lowered contents of polyunsaturated FAs. This also applies to caput epididymis, while EFAD sensitivity is regained in cauda epididymis, where the desaturases are upregulated. The FA profile of epididymal spermatozoa is increasingly affected by EFAD during the transport from testis to cauda epididymis. Furthermore, a significant increase in the number of abnormal spermatozoa is observed in cauda epididymis.     

A training course on food hygiene for butchers: measuring its effectiveness through microbiological analysis and the use of an inspection checklist

The effectiveness of food hygiene training for a group of retail butchers was evaluated with the aim of verifying whether the butchers modified their behavior in the light of knowledge gained and whether their acquired knowledge or behavior change was sustained over a period of time. Microbiological analysis (enumeration of mesophilic and coliform bacteria and Escherichia coli) of a raw semiprocessed product (stuffed rolled beef) was conducted, and an inspection checklist was issued before the training course (T0). Initial results were later compared with results obtained 1 month (T1) and 6 months (T6) after the training. The checklist comprised 89 items classified into five categories: A, approved suppliers and product reception; B, storage conditions and temperature control; C, flow process, food handling procedures, and conditions of the window display unit; D, facility design and proper cleaning and sanitizing of equipment, utensils, and work surfaces; and E, pest control system, water supply control, and garbage disposal. The inspection results were recorded as "yes" or "no" for each item. Compliance with food safety procedures was recorded as the percentage of "yes" answers. The bacterial counts were significantly higher at T0. At T6, there was no significant increase in bacterial counts. There was a significant improvement in food safety practices at T1 and T6 compared with T0 for all categories. When comparing T0 and T1, the largest increases in the compliance scores were seen within categories C and D. No significant decrease in scores for compliance with food safety practices was observed at T6. Supervision and refresher activities may be necessary to maintain behavioral changes for a longer period of time.     

Effects of photoperiod during the dry period on prolactin, prolactin receptor, and milk production of dairy cows

Cows exposed to short day photoperiod during the dry period produce significantly more milk in their subsequent lactation than cows exposed to long days. The mechanism(s) underlying this effect are unknown. Because concentrations of prolactin (PRL) in circulation are consistently affected by changes in photoperiod, we hypothesized that alterations in the prolactin axis and sensitivity of the mammary gland to prolactin signaling may mediate photoperiodic effects in dry cows. The objective of this study was to determine the effects of exposure to different lengths of daylight during the dry period on circulating PRL and PRL receptor (PRL-R) mRNA expression in lymphocytes and mammary tissue during the transition to lactation. Multiparous Holstein cows were dried off 62 d before calving and assigned to long day (16 h light: 8 h dark) or short day photoperiod (8 h light: 16 h dark). During the dry period, PRL and PRL-R mRNA were analyzed biweekly in plasma and lymphocytes, respectively. Expression of PRL-R mRNA was assessed in mammary biopsies during the dry and periparturient periods. Dry matter intake (DMI) was recorded through 21 d of lactation, and milk yield was recorded until 120 d in milk. Short day photoperiod was associated with reduced PRL, whereas milk yield and expression of PRL-R mRNA in lymphocytes and mammary tissue were increased. Cows on short days had higher DMI during the dry period but did not differ in DMI after parturition. These data support the concept that greater responsiveness and sensitivity to PRL during transition to lactation may be associated with an increase in subsequent milk yield.     

Izumoring: a novel and complete strategy for bioproduction of rare sugars

Starch, whey or hemicellulosic waste can be used as a raw material for the industrial production of rare sugars. D-glucose from starch, whey and hemicellulose, D-galactose from whey, and D-xylose from hemicellulose are the main starting monosaccharides for production of rare sugars. We can produce all monosaccharides; tetroses, pentoses and hexoses, from these raw materials. This is achieved by using D-tagatose 3-epimerase, aldose isomerase, aldose reductase, and oxidoreductase enzymes or whole cells as biocatalysts. Bioproduction strategies for all rare sugars are illustrated using ring form structures given the name Izumoring.