High-resolution protein structure determination by serial femtosecond crystallography

Structure determination of proteins and other macromolecules has historically required the growth of high-quality crystals sufficiently large to diffract x-rays efficiently while withstanding radiation damage. We applied serial femtosecond crystallography (SFX) using an x-ray free-electron laser (XFEL) to obtain high-resolution structural information from microcrystals (less than 1 micrometer by 1 micrometer by 3 micrometers) of the well-characterized model protein lysozyme. The agreement with synchrotron data demonstrates the immediate relevance of SFX for analyzing the structure of the large group of difficult-to-crystallize molecules.     

Factors influencing pregnancy rate and late embryonic loss in dairy cattle.

This trial was conducted on three Hungarian dairy farms between March 2003 and April 2004. The aims of this study were to examine the effect of milk production and milk contents, the day after calving (DAC) and the sire for the pregnancy rate (PR) and late embryonic loss (LEL) in dairies. Five thousand three hundred and eighty insemination was carried out in this period. Pregnancies (n = 1969) were detected by BioPryn test from 3919 blood samples between 30-36 days post insemination (PI). Retention of pregnancy was determined by rectal palpation on Day 60. LEL has been determined by the optical density (OD) of blood samples/cutoff ratio and the serum progesterone (P4) concentration 30-36 days PI. According to serum progesterone concentration the authors predicted a presumed or possible embryonic loss or maintenance of the pregnancy. The efficiency of the artificial inseminations (AI) was significantly lower 60 days within post-partum. Significant positive correlation was found between the result of early and late PR and DAC and the milk protein/fat ratio. Significant negative correlation was detected between the early and late PR the milk protein and fat content. A later stage (60 days PI) of pregnancy the milk production has a negative effect for retaining of pregnancy. More data would be necessary to state that sires can influence the reproduction ability of their progenies. This is a really important issue, because the AI bulls have a great impact on the herds and the one-way selection may cause decreased fertility on breed level as well.     

Physiological Variation in the Genus Meloidogyne as Determined by Differential Hosts

Four species of root-knot nematodes, namely Meloidogyne incognita (Kofoid et White) Chitwood, M. javanica (Treub) Chitwood, M. arenaria (Neal) Chitwood, and M. hapla Chitwood have a wide host-range and are widely distributed throughout the world. For each of the species there are certain non-hosts and these are different for each species, thus making it possible, through the use of differential hosts, to identify the species as well as detect pathogenic variations within a species. A knowledge of the nature and extent of this variation is needed for the development of varieties which possess a broad base for resistance.     

Performance of Pinus elliottii, Pinus caribaea, their F1, F2 and backcross hybrids and Pinus taeda to 10 years in the Mesopotamia region, Argentina

Performances of Pinus taxa were studied to 10 years of age in two trials in each of Misiones and Entre Ríos provinces across the Mesopotamia region of Argentina. Taxa comprised 22 populations from sources in Argentina, Australia, Brazil and Zimbabwe including Pinus elliottii var. elliottii (Pee), Pinus caribaea var. hondurensis (Pch), their four, inter-specific hybrids (F₁, F₂ and backcrosses from F₁ to Pch and to Pee—all as broadly based bulks); other Pee and Pinus taeda (Pt) comprised narrower or unspecified bulks. Variable numbers of taxa were missing at each site. Mean survival across sites at age 10 years ranged 53.2–91.3% averaging 74.2%. Analysis of variance of plot means indicated population effect was statistically significant (p < 0.05) for all or most growth and quality traits at all sites. However, significant differences from the nominated check seedlot at the Entre Ríos sites (Pee, Australia) were extremely rare, while quite common at the northern, Misiones sites (check seedlot a Pt population). In the Misiones trials, F₁, F₂ and both backcross hybrids showed better stem straightness than Pee and Pt from Argentina, generally with statistically significant differences (p < 0.05). Pt showed lowest forking scores (desirable). Taxon × environment interaction was statistically significant (p < 0.01) for growth traits only (p > 0.05). However, this interaction contributed an average of only 34.1% of the taxon variance suggesting a lack of practical importance. Taxa most suitable for deployment in the Mesopotamia region, Argentina are suggested.     

Improving the Reproductive Efficiency by Zoo-Technical Methods at a Dairy Farm

This trial was conducted on a Hungarian dairy farm between July 2001 and December 2004. The objective of this work was to improve the reproductive efficiency with relatively high milk production. At the beginning of this trial blood and fodder samples were taken for checking the metabolic status of the animals in order to determine their health condition. The nutritive value of the daily ration for all groups met with the requirements of the Hungarian National Standard and almost all serum metabolic parameters differed between the milking and pregnant animals. Early pregnancy detection (by ultrasound) and ovulation synchronizing methods were introduced to optimize reproductive performance. The oestrus cycle was also checked by ultrasound and open cows were treated by the appropriate method in order to inseminate them as soon as possible. Efficiency of artificial insemination (AI) followed by a single prostaglandin F2alpha (PGF2alpha) and Ovsynch treatment was similar (30.8% and 29%) and less effective than AI after natural heat detection (37.1%). Provsynch (Pre-synch followed by Ovsynch) was the most effective ovulation synchronization method (conception rate=42.6%; p<0.01). Although milk production increased between 2002 and 2004 by approximately 600 kg per cow, the calving period decreased by 20 days and the number of AIs per pregnancy was also improved (0.8 AI per conception). These findings are really beneficial economically because the decrease in the calving interval returns profit for the dairy farms; one open day costs euro2.5/cow. Economical analysis showed a high profit ratio from the reproductive 'investment' on this farm. Every invested euro yielded approximately euro10.     

Postpartum reproduction in protein restricted beef cows: effect on the hypothalamic-pituitary-ovarian axis

The influence of dietary CP on circulating LH and anterior pituitary and hypothalamic function was examined. In Exp. 1, 28 cows were randomly assigned to four treatment groups: adequate CP (ADQ; .96 kg/d) or deficient CP (DEF; .32 kg/d) beginning at 90, 60 and 30 d before parturition and continued at a 33% increase in feed consumption after parturition. Cows were bled at 15-min intervals for 8 h on d 20, 40 and 60 after parturition. Pituitaries were collected on d 62 to analyze GnRH receptor numbers and gonadotropin content. Frequency of pulsatile LH release increased (P less than .05) from 20 to 60 d in ADQ cows. Basal and mean LH were not affected (P greater than .10) by CP restriction or by days after parturition. Crude protein did not affect pituitary GnRH receptors (P greater than .10), but it did affect pituitary LH content, FSH content and FSH concentration (P less than .05). In Exp. 2, 28 cows were assigned to treatment groups as in Exp. 1. All cows were challenged with GnRH (.22 micrograms/kg BW) at 20, 40 and 60 d after parturition and were bled every 30 min for 6 h. Responsiveness to GnRH increased with increased time after parturition (P less than .07). Deficient CP decreased GnRH-induced LH release (P less than .05). In Exp. 3, 12 cows were randomly assigned to ADQ or DEF CP beginning 120 d before parturition. All cows received 1 mg estradiol-17 beta (E2) on d 19, 39 and 59 after parturition and were bled every 30 min for 14 h beginning 14 h following E2. Response to E2 was unaffected by CP restriction (P greater than .10), whereas time to E2-induced LH peak decreased as time after parturition increased in ADQ cows (P less than .05). Results suggest that delayed return to estrus in CP-deficient postpartum beef cows might be due to reduced gonadotropin release from the anterior pituitary and decreased anterior pituitary responsiveness to GnRH.