Separation of D- and L-amino acids by ion exchange column chromatography in the form of alanyl dipeptides

Summary A method of ion exchange column chromatography was developed for the determination of D- and L-amino acids in the form of diastereomeric dipeptide. First the protein containing samples were hydrolyzed with 6 molar hydrochloric acid, then the single amino acids were separated in an LKB automated amino acid analyzer with the LKB fraction collector. Following lyophilization, the single amino acids were transformed into alanyl dipeptides with tertiary-butyloxycarbonil-L-alanine-N-hydroxy-succinimide (t-BOC-L-Ala-ONSu) active ester. The alanyl dipeptides were easily separated from one another and the initial amino acids. Determination of the D- and L-amino acids in this form is relatively accurate and reproducible but takes some time (33–38 min). Accuracy of the determination is satisfactory. The coefficient of variation amounts to 3–5%. The use of the method is suggested to laboratories having an amino acid analyzer and wish to determine D-and L-amino acids in synthetic-amino acids complements, peptides or natural materials.     

A new fourier transform approach for protein coding measure based on the format of the Z curve

MOTIVATION: At the core of most protein gene-finding algorithms are the coding measures used to make a decision on coding/non-coding. Of the protein coding measures, the Fourier measure is one of the most important. However, due to the limited length of the windows usually used, the accuracy of the measure is not satisfactory. This paper is devoted to improving the accuracy by lengthening the sequence to amplify the periodicity of 3 in the coding regions. RESULTS: A new algorithm is presented called the lengthen-shuffle Fourier transform algorithm. For the same window length, the percentage accuracy of the new algorithm is 6-7% higher than that of the ordinary Fourier transform algorithm. The resulting percentage accuracy (average of specificity and sensitivity) of the new measure is 84.9% for the window length 162 bp. AVAILABILITY: The program is available on request fromC.- T. Zhang. Contact: ctzhang@tju.edu.cn      

Assessment of the efficiency of a hand-held suction device for sampling spiders: improved density estimation or oversampling?

In the study a comparison is made between the results of two sampling procedures, both based on the application of the same hand-held suction apparatus. Sampling was aimed at spiders, and was carried out on two alfalfa fields. In the first method suction sampling was applied to an enclosure of 0.48 m². The enclosed area was sampled intensively, which was facilitated by the removal of the vegetation. The second method was a transect sampling procedure during which the suction apparatus with a 0.01 m² nozzle was applied to single unenclosed sampling points 1 m apart. A linear series of 48 such subsamples comprised a transect, thus the total area covered in a transect equalled the area of the enclosure. In the transect samples three times more spiders were caught than in the enclosures. This result was consistent on different occasions and at both fields. This basic trend was found in all spider families that were present in significant numbers in the samples. Species composition in the samples collected by the two methods was similar, and species abundance ranks were highly correlated across dates. We propose that an “edge effect” can explain higher catches in transect samples. This edge effect is caused by lateral suction at the edges, which inflated the number of animals caught in the transect application.     

Ontogenic Development of Corticotrophs in Fetal Buffalo (Bubalus Bubalis) Pituitary Gland

To evaluate the subpopulation of corticotrophs in developing buffalo (Bubalus bubalis) fetus, pituitary glands were recovered (n=6 per group) from late first, second and third gestational female buffalo dams. The corticotrophs were identified by using specific antibodies against proopiomelanocortin (POMC) and adrenocorticotrophic hormone (ACTH) through immunohistochemistry. There was a significant (P≤0.05) increase of immunoreactive (ir) ir-ACTH cells during late 2^nd trimester while, ir-POMC cells were more (P≤0.05) at late 3^rd trimester of gestation as compared to other age groups. The quantity of co-localized cells for POMC and ACTH was significantly (P≤0.05) greater at the end of 1^st gestation rather than 2^nd and 3^rd gestational fetal adenohypophyseal cells. This study is the first to demonstrate co-localization of POMC+ACTH and the affect of gestational age on the expression of these cells in buffalo fetus adenohypophysis.Key words: buffalo, corticotrophs, fetus, pituitary gland, immunohistochemistry     

Changes in sperm stores, ejaculate size, fertilization success, and sexual motivation over repeated matings in the common toad, Bufo bufo (Anura: Bufonidae)

Fertilization is of central importance in the determination of reproductive success for both males and females. In species where males have the chance to mate repeatedly within a short period of time, sperm stocks may become depleted and males may have to carefully economize on their sperm reserves. Also, intensive intrasexual competition for females and repeated matings may lead to exhaustion on the behavioural level. To determine whether the reproductive potential of males is limited and if such a limitation is due to behavioural exhaustion or sperm depletion, we experimentally investigated changes in sperm stores, sperm expenditure, fertilization success, and sexual motivation over three repeated matings in the common toad, Bufo bufo , where the breeding season is short and sequential polygyny occurs. At the end of the breeding season, the number of sperm stored in the testes of males mated repeatedly was close to 50% lower than in testes of unmated males. Ejaculate size, which was estimated by applying a novel method allowing direct quantification, decreased by 88% from first to third matings. We also observed a drop in fertilization success from the first two to third matings by 65%, which was largest in males that had started the reproductive season in bad body condition. Some of these males also showed a decreased interest in females in the third mating round. Our results suggest that sperm depletion and loss of sexual motivation may together set a limit to the reproductive potential of common toad males. The present study draws attention to a limitation in reproductive potential, which may occur more often than currently anticipated and has the potential to strongly influence several aspects of reproductive behaviour.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 361–371.     

The purification of a bovine kidney enzyme which cleaves melanocyte-stimulating hormone-release inhibiting factor.

An enzyme which catalyzes the hydrolysis of L-prolyl-L-leucylglycinamide, the factor which inhibits the release of melanocyte-stimulating hormone, was purified 189-fold from bovine kidney in a 5% yield. The molecular weight of the enzyme on gel filtration was estimated to be 300 000 and its isoelectric point was found to be pH 4.1. The single component seen on sodium dodecyl sulphate-gel electrophoresis was estimated to have a molecular weight of 56 000, indicating that the native enzyme may be a pentamer or hexamer. The enzyme could clearly be distinguished from other prolyl-cleaving enzymes.     

Purification and partial characterisation of a bovine kidney aminotripeptidase (capable of cleaving prolyl-glycylglycine)

Summary An enzyme was purified 163-fold in an 8.2% yield from bovine kidney. The specific activities of the pure preparation against L-prolyl glycylglycine and L-alanyl glycylglycine were found to be 244.5 and 578 , Moles substrate hydrolyzed per min per mg protein respectively. The molecular weight of the enzyme was estimated on gel filtration to be 55,000. The isoelectric point was recorded to be pH 5.2. A preliminary study of substrate specificity showed that the enzyme preferentially hydrolyzed tripeptides of the type X-glycylglycine. The enzyme was tentatively identified as a tripeptide aminopeptidase ( aminoacyldipeptide hydrolase, EC 3.4.11.4).Abbreviations DEAE- diethylaminoethyl- - QAE- quarternaryaminoethyl- - SP- sulphopropyl- - CM- carboxymethyl- - B_z- -N-benzoyl- - Z- -N-benzyloxycarbonyl-