Site-directed PEGylation of trichosanthin retained its anti-HIV activity with reduced potency in vitro

Trichosanthin (TCS) was the first ribosome inactivating protein found to possess anti-HIV-1 activity. Phase I/II clinical trial of this compound had been done. Antigenicity and short plasma half-life were the major side effects preventing further clinical trial. Modification of TCS is therefore necessary to revive the interest to develop this compound as an anti-HIV agent. Three potential antigenic sites (Ser-7, Lys-173, and Gln-219) were identified by computer modeling. Through site-directed mutagenesis, these three antigenic amino acids were mutated to a cysteine residue resulting in 3 TCS mutants, namely S7C, K173C, and Q219C. These mutants were further coupled to polyethylene glycol with a molecular size of 20 kDa (PEG) via the cysteine residue. This produced another three TCS derivatives, namely PEG(20)k-S7C, PEG(20)k-K173C, and PEG(20)k-Q219C. PEGylation had been widely used recently to decrease immunogenicity by masking the antigenic sites and prolong plasma half-life by expanding the molecular size. The in vitro anti-HIV-1 activity of these mutants and derivatives was tested. Results showed that the anti-HIV-1 activity of S7C, K173C, and Q219C was decreased by about 1.5- to 5.5-fold with slightly lower cytotoxicity. On the other hand, PEGylation produced larger decrease (20- to 30-fold) in anti-HIV activity. Cytotoxicity was, however, weakened only slightly by about 3-fold. The in vitro study showed that the anti-HIV activity of PEGylated TCS was retained with reduced potency. The in vivo activity is expected to have only slightly changed due to other beneficial effects like prolonged half-life. (C) 2004 Elsevier Inc. All rights reserved.

Effects of irradiation on formaldehyde concentration and nutritional changes of formalin treated fish, Pampus chinensis

Formaldehyde is a very reactive compound capable of interacting with many functional groups of proteins including intermolecular and intramolecular cross-links of the molecules. The formation of cross-linking bonds may induce conformational change in proteins that favor further interaction of functional and hydrophobic groups. Formaldehyde which has been using illegally as a chemical preservative by some fish traders in our country. A study was carried out to determine the effects of irradiation (1.5 KGy) on formaldehyde concentration and nutritional (protein and lipid) changes of formalin (37% formaldehyde) treated fish (fresh) samples and found that the concentration of formaldehyde both in treated samples (0.37% formalin and 0.37% formalin with 1.5 KGy irradiation) were 37.0 µg/gm and 36.75 µg/gm. On the other hand, the amount of protein and lipid in treated samples before radiation (14.56% and 3.49%) and after radiation (14.15% and 3.25%). That means, radiation has no effect on the change of protein, lipid and formaldehyde.

The genetic diversity of Acipenser stellatus populations in the Caspian Sea was studied using microsatellite technique

A total of 361 caudal fin samples were collected from adult A. stellatus specimens caught in the north Caspian Sea, including specimens from Kazakhstan (Ural River), Russia (Volga River), Azerbaijan (Kura River), specimens caught in the south Caspian Sea including specimens from Fishery Zone 1 (from Astara to Anzali), Fishery Zone 2 (from Anzali to Ramsar), Fishery Zone 3 (from Nowshahr to Babolsar), Fishery Zone 4 (from Miyankaleh to Gomishan) as well as from specimens caught in Turkmenistan (all specimens were collected during the sturgeon stock assessment survey). About 2 g of fin tissue was removed from each caudal fin sample, stored in 96% ethyl alcohol and transferred to the genetic laboratory of the International Sturgeon Research Institute. Genomic DNA was extracted using phenol-chloroform method. The quality and quantity of DNA was assessed using 1% Agarose gel electrophoresis and Polymerase Chain Reaction (PCR) was conducted on the target DNA using 15 paired microsatellite primer. PCR products were electrophoresed on polyacrylamide gels (6%) that were stained using silver nitrate. Electrophoretic patterns and DNA bands were analyzed with BioCapt software. Allele count and frequency, genetic diversity, expected heterozygosity and observed heterozygosity allele number, and the effective allele number, genetic similarity and genetic distance, FST and RST were calculated. The Hardy Wienberg Equilibrium based on X2 and Analysis of Molecular Variance (AMOVA) at 10% confidence level was calculated using the Gene Alex software. Dendrogram for genetic distances and identities were calculated using TFPGA program for any level of the hierarchy. It is evident from the results obtained that the 15 paired primers studied, polymorphism was observed in 10 pairs in 12 loci, while one locus did not produce DNA bands. Mean allele number was 13.6. Mean observed and expected heterozygosity was 0.86 and 0.642, respectively. It was also seen that specimens from all regions were not in Hardy Wienberg Equilibrium in most of the loci (P≤0.001). Highest Fst (0.063) was observed when comparing specimens from Fishery Zone 2 and Fishery Zone 4 (Nm=3.7) and lowest FST (0.028) was observed when comparing specimens from the Volga River and those from the Ural River (8.7). Significant differences (P<0.01) were observed between RST recorded in the specimens studied. Highest genetic distance (0.604) and lowest genetic resemblance (0.547) were observed between specimens from Fishery zones 2 and 4. Lowest genetic distance (0.311) and highest genetic resemblance (0.733) was observed between specimens from Turkmenistan and specimens from Fishery zone 1. Based on the genetic dendrogeram tree derived by applying UPGMA algorithm, A. stellatus specimens from Fishery zone 2 or in other words specimens from the Sepidrud River belong to one cluster which divides into two clusters, one of which includes specimens from Fishery zones 1, 3 and 4 and specimens from Turkmenistan while the other cluster includes specimens from Ural, Volga and Kura Rivers. It is thus evident that the main population of this species belongs to the Sepidrud River. Results obtained from the present study show that at least eight different populations of A. stellatus are found in the north and south Caspian Sea, four of which are known populations including the Ural River population, the Volga River population, the Kura River population and the Sepidrud River populations. The four other populations identified belonging to Fishery zones 1, 3, and 4 and to Turkmenistan are most probably late or early spawners of the spring run and autumn run of each of the major rivers mentioned. Specific markers were also identified for each of the populations identified. The Ural River population can be identified using primers Spl-68, 54b and Spl-104, 163 170, 173, the Volga River population can be identified using primers LS-54b and Spl-104, 170, 173 113a and similarly the population from the Kura River can be identified using primers LS-34, 54b and Spl-163, 173 and that from the Sepidrud River can be identified using primers LS-19, 34, 54b and Spl-105, 113b. This study gives evidence of the presence of different populations of this species and calls for serious measures to be taken to protect the genetic stocks of these populations. Considering that the population of A. stellatus in Fishery zone 2 is an independent population of the Sepidrud River in the Gilan Province, the catch of these fishes in the region needs to be controlled and regulated in order to restore the declining stocks of this species.

中国翼手类一属、种新纪录

记述了采自中国云南西部和西北部的无尾果蝠属Megaerops及其两个种:泰国无尾果蝠M.niphanae和无尾果蝠M.ecaudatus为翼手类中国属、种薪纪录.标本收藏于中国科学院昆明动物研究所.

Population dynamics and management of yellowfin tuna (Thunnus albacares) of the Oman Sea

The Yellowfin tuna was caught more than all other species in the southern waters of Iran (24000 tons in 1998). In order to come up with the responsible fishing pattern, there was a need to identify some of the biological characteristics and population dynamic parameters. This thesis was the first which covered the whole Yellowfin tuna distribution in the Oman Sea, included the fishing grounds of Berris, Ramin, Chabahar, Pozm and Jask. The data during 1998-99 from different fishing grounds were polled. Based on the exponential relationship between length and weight in the size range 38-173 Cm, the relationship (W=aL^ b) was calculated as W=0.000012L ^ 3.0831). The mean fork length,head length,girth and weight were calculated respectively 84.15 Cm, 23 Cm, 53 Cm, and 11828 g. Length infinity was estimated 189 Cm with growth parameters of 0.42 per year. Growth performance index was 4.18 which was in agreement with the findngs of the other studies in the Indian and Pacific Oceans. The mortality parameters and exploitation rate were estimated as below: Z = 1.75-1.85 M=0.6 F=1.25 E=0.68 Occurence of empty stomach was high (60%) in the speciemens obtained from the Oman Sea. Purpleback flying squid (Sthenoteuthis oualaniensis) was the most dominant prey species observed in the study (57% in females and 60% in males), occurrence of teleost fishes were found to be the second (38% in males and 42% in females). Crabs also were identified in the specimens(1-2%). The study on sex ratio indicated that males were predominant at all sizes above 120 Cm fork length. 50.82% of specimens were males and 49.18% females. The monthly gonadosomatic index was deriven higher values during January to June which could be indicated as spawning period.

Morphine withdrawal affects both delayed-escape behaviour in Morris water maze and hippocampal NR2A/2B expression ratio

Repeated low-dose morphine treatment facilitates delayed-escape behaviour of hippocampus-dependent Morris water maze and morphine withdrawal influences hippocampal NMDA receptor-dependent synaptic plasticity. Here, we examined whether and how morphine wit

金翅雀的繁殖生态

2005年3月-7月对南充市金翅雀的繁殖生态进行了研究.结果表明:金翅雀3月开始繁殖;雌雄共同筑巢,营巢期4-5d;窝卵数:3.67±0.58(n=3),孵化率63.64%;雌孵卵,孵卵期12-13d,孵卵期间食物由雄鸟提供;双亲共同育雏,育雏期14-15d.雏鸟生长曲线呈“S”型.

湖南产尖吻蝮蛇蛇毒出血毒素性质和溶液构象的研究

经sephadex G-75凝胶过滤、QAE-Sephadex A-50和CM-Sephadex C-25离子交换步骤,从湖南产尖吻蝮（Deinagkistrodon acutus）蛇毒中纯化出两个出血毒素,被命名为HaHT-1和HaHT-2。用聚丙烯酰胺凝胶电泳（PAGE）和SDS-聚丙烯酰胺凝胶电泳鉴定呈单一的蛋白染色带。两个出血毒素的分子量相同,均为 23.5kDa。等电聚焦电泳测定它们的等电点分别为5.6和5.2。HaHT-1和HaHT-2均具有较强的出血活性（MHD分别为0.5和0.8μg）,都有酪蛋白水解活力,无精氨酸酯酶、胆碱酯酶和磷脂酶A_2活力。用CD谱测定HaHT-l和HaHT-2的溶液构象,HaHT-1的α-螺旋、β-折叠和无规卷曲分别为36.9％、35.5％和27.6％,而HaHT-2的α-螺旋、β-折叠和无规卷曲分别为23.4％、31.3％和45.3％。pH的变化对它们构象有影响,在pH2-11范围内,酸性比碱性大。随着酸性或碱性的增加其α-螺旋含量减少,无规卷曲增加,β-折叠结构变化不大。