Anti-HIV-1 property of trichosanthin correlates with its ribosome inactivating activity

Trichosanthin (TCS) is a type I ribosome inactivating (RI) protein possessing anti-tumor and antiviral activity, including human immunodeficiency virus (HIV). The mechanism of these actions is not entirely clear, but is generally attributed to its RI property. In order to study the relationship between the anti-HIV-1 activity of TCS and its RI activity, three TCS mutants with different RI activities were constructed by using site-directed mutagenesis. The anti-HIV-1 activities of the three mutants were tested in vitro. Results showed that two TCS mutants, namely TCSM((120-123)), TCSE160A/E189A, with the greatest decrease in RI activity, lost almost all of the anti-HIV activity and cytopathic effect. Another mutant TCSR122G, which exhibited a 160-fold decrease in RI activity, retained some anti-HIV activity. The results from this study suggested that RI activity of TCS may have significant contribution to its anti-HIV-1 property. (C) 2002 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

Independency of anti-HIV-1 activity from ribosome-inactivating activity of trichosanthin

Trichosanthin (TCS) is a type I ribosome-inactivating (RI) protein possessing multiple biological and pharmacological activities. Its major action is inhibition of human immunodeficiency virus (HIV) replication but the mechanism is still elusive. All evidences showed that this action is related to its RI activity. Previous studies found that TCS mutants with reduced RI activity simultaneously lost some anti-HIV activity. In this study, an exception was demonstrated by two TCS mutants retaining almost all RI activity but were devoid of anti-HIV-1 activity. Five mutants were constructed by using site-directed mutagenesis with either deletion or addition of amino acids to the C-terminal sequence. Results showed that the RI activity of mutants with C-terminal deletion mutants (TCSC2, TCSC4, and TCSC14) decreased by 1.2-3.3-fold with parallel downshifting of its anti-HIV-1 activity (1.4-4.8-fold). Another two mutants, TCSC19aa and TCSKDEL having 19 amino acid extension and a KDEL signal sequence added to the C-terminal sequence, retained all RI activity but subsequently lost most of the anti-HIV-1 activity. These findings suggested that ribosome inactivation alone might not be adequate to explain the anti-HIV action of TCS. (C) 2003 Elsevier Science (USA). All rights reserved.

Notes on the construction of 12 cu m ferrocement maturation tank for prawn broodstock

The study aimed to develop an economical, rational and easy method of constructing ferrocement tank suitable for P. monodon maturation.

Effect of different sex ratios of ablated wild-stock Penaeus monodon Fabricius on maturation, fecundity and hatching rates

The study was conducted to determine other sex ratios of ablated wild-stock Penaeus monodon other than the most commonly practiced 1 male: 1 female. Four different sex ratios, 0:1, 1:1, 1:2, and 1:4 male: female were tested in four 4m diameter circular tanks for a period of 55 days. During the first run the 1 male: 2 females ratio gave: (a) the highest percentage of first (42.20%), second (30.00%) and third (33.33%) spawning; and (b) the highest total and average fecundity (3.9 million eggs and 300,692 eggs, respectively). The 1 male: 2 females ratio is recommended on the basis of highest percentage for the first, second and third spawners, total and average fecundity.

Survival, maturation, fecundity and hatching rates of unablated and ablated Penaeus indicus H.M. Edwards from brackishwater ponds

Penaeus indicus harvested after three months of rearing in brackishwater ponds and averaging 6.9 g for females and 5.6 g for males were stocked in two 12 cu m flowthrough ferrocement tanks at 240 females and 200 males per tank. The females were ablated on one eyestalk in one tank and remained unablated in the other tank; all males were unablated. Ablated females spawned up to 7 times per female; unablated females spawned up to only 3 times during the two month duration of the experiment. Ablated females produced a total number of 17.5 x 10⁶ eggs, 6.6 x 10⁶ nauplii, and an average of 23,480 eggs/spawning and 37.8% hatching rate from a total of 757 spawnings. Unablated females produced a total of 2.0 x 10⁶ eggs, 1.1 x 10⁶ nauplii, and an average of 26.990 eggs/spawning and 53.9% hatching rate from a total of 74 spawnings. Survival of ablated females was 53.5% compared to 69.4% for unablated females; males in both tanks averaged more than 90% survival.

Bighead carp - its maturation and ovulation

The paper investigates the effects of intraperitoneal injections of LHRH-a and domperidone (DOM), given singly or in combination at two injections, on oocyte maturation and spawning in bighead carp, Aristichthys nobilis.

CORRELATION OF ZONA-BINDING WITH OOCYTE MATURATION AND SPERM MOTILITY IN RHESUS-MONKEYS BY HEMIZONA ASSAY

The hemizona assay (HZA) in Rhesus monkeys was employed to study the correlation of zona-binding ability with sperm motility or with naturally developing oocytes at various maturational stages. Oocytes from unstimulated ovaries were retrieved within 2 hr from monkeys sacrificed for vaccine production (in reproductive season, but with their menstrual cycles not determined). Oocytes were divided into four groups based on their morphological maturation: 1) Oocytes surrounded by more than one cumulus layer (MC); 2) Oocytes retaining intact germinal vesicle nuclei (GV); 3) Oocytes with germinal vesicle breakdown showing distinct perivitelline space (PVS); and 4) Oocytes extruding the first polar body (PB1). The mean numbers of sperm bound to hemizona for PBI, PVS, GV, and MC groups were 132.9 +/- 12.0, 71.5 +/- 10.1, 36.1 +/- 4.0, and 20.1 +/- 2.9 (Mean +/- SE), respectively. The four groups showed significant differences from each other in sperm/egg binding ability (P < 0.01). The number of bound sperm significantly increased with oocyte maturation. The present study also showed that zona-binding ability was also affected by sperm motility. For sperm with 67.7% motility and sperm with 31.2% motility, the average numbers of bound sperm were 43.5 +/- 2.2 and 25.3 +/- 2.9 (Mean +/- SE), respectively. There was significantly higher binding ability for sperm with higher motility (P < 0.01). The results suggest that: 1)The rhesus monkey model can serve as a very sensitive model for studying sperm/egg interaction by HZA; 2) Sperm motility positively correlated with sperm/egg binding; and 3) Sperm/egg binding ability increases with oocyte maturation. The binding ability is highest when oocytes matured to the PB1 stage, which is also the best opportunity for fertilization. This is strong evidence for the ''zona maturation'' hypothesis. (C) 1994 Wiley-Liss, Inc.

Study on biological characteristics, sexual maturation and reproduction of Liza abu in the water of Khozestan Province

As the most of the fish resources are known and exploited, protecting their generation is of the greatest importance. Aquaculture is one of the efficient procedures in protecting and reviving fish resources and knowing about the reproductive cycle and gonads development has an important role in approaching this aim. Liza abu belongs to the family Mugilidae that according to its resistance to the environmental condition and its fast growth , can be introduced as a fish with economical value. As there is no scientific data on the reproductive biology of this species , study on the reproductive biology and gonad development is considered as the aim of this research . For this purpose , 360 samples of this species were investigated during the period from February 2007 to January 2008 in Khozestan Province . After studing morphological and histological characteristics of gonad specimen , they were prepared through histological method. Samples were prepared through usual histological method and studied under light microscope. According to the results, the maturity stages of male and female Liza abu were separated to six different successive stages. In ovaries , these stages were as follow : In stage І, the oocytes were small , this stage was observed from July to October . In stage ІІ, considerable growth was observed in the oocytes . This stage was observed from October to January . In stage III, due to vitellogenesis, the maximum growth was observed and three layers of theca, granullosa and follicle cells were visible. This stage was observed during January and February . In stage IV, migration of germinal vesicle was observed and due to hydration of the oocytes , their diameter was increased. The ovaries were yellowish and in maximum size and ovules could be easily observed with naked-eye . This stage was observed in February and March . In stage V, spawning occured. This stage was observed in April . In stage VI, ovaries consisted of immature and atretic oocytes and also empty follicles. This stage was observed in May and June. In testes , these stages were as follow : In stage I , the testes were small in size and contained the spermatogonia which were the only cellular components.This stage was observed in August and September . In stage II (maturing virgin ) , the spermatogonia and the primary spermatocytes were visible. This stage was observed in October . In stage III (developing), intensive spermatogenesis was occured and the primary and the secondary spermatocytes were the most visible cells during this stage .This stage was observed from November to January. In stage IV(developed), cells of all stages of spermatogenesis could be seen but the secondary spermatocytes and spermatids were in large number. This stage was observed from January to March. In stage V , the testes were filled with sperms. This stage was observed in March and April .In stage VI, residual spermatozoa and the spermatogonia were visible in the testes. This stage was observed from May to August. According to cyclic changes in GSI, sexual maturation in breeding begins in January and spawning occurs in April. The ova diameter ranged from 30.75 μ in stage I to 472.19 μ in stage IV. In this study , the sex ratio was 1:2.7, and male and female percentage were 27.02% and 72.98% respectively. This means that females predominate males. In this study absolute fecundity was calculated and changing between 30805.44 to 431247.3 was observed and absolute fecundity was calculated 111275.3 in average.

Amino acid requirements for maturation of rhesus monkey (Macacca mulatta) oocytes in culture

This study evaluated the effects of different amino acid formulations on supporting meiotic and cytoplasmic maturation of rhesus monkey (Macacca mulatta) oocytes in vitro. Five hundred and forty-six cumulus-oocyte complexes (COCs) aspirated from unstimulated adult monkey follicles (greater than or equal to 1000 mum in diameter) were cultured in either modified Connaught Medical Research Laboratories 1066 medium (mCMRL-1066) or in one of eight chemically defined media (modified basic medium 5 supplemented with 5.5 mmol glucose l(-1), 0.003 mmol pantothenic acid l(-1) and different amino acid formulations) as below: (1) modified basic medium 5 (mBM5) containing no amino acid; (2) mBM5 + 0.2 mmol glutamine l(-1); (3) mBM5 + 11 amino acids from hamster embryo culture medium 6 (HECM-6) (11 AA); (4) mBM5 + Eagle's non-essential amino acids (NEA); (5) mBM5 + NEA + 0.2 mmol glutamine l(-1); (6) mBM5 + Eagle's essential amino acids (EA) without glutamine; (7) mBM5 + EA + 0.2 mmol glutamine l(-1); (8) mBM5 + Eagle's 20 amino acids (20 AA) + 0.2 mmol glutamine l(-1); and (9) mCMRL-1066 (control). All media contained FSH, LH, oestradiol and progesterone. After maturation, mature oocytes were subjected to the same fertilization and embryo culture procedures. COCs matured in treatment 5 had greater potential to progress to metaphase II (66%; P < 0.05) than did those in treatments 1 (37.3%), 2 (48.3%)f 3 (41%), 6 (41%) and 9 (43%). Oocytes matured in treatment 8 had the best morula (53%) and blastocyst (18%) developmental responses (P<0.05). The lowest (P<0.05) morula and blastocyst developmental responses were obtained from COCs matured in treatments 1 (0%) and 6 (8%). The other media supported intermediate embryonic development (range 11-38% of morula and blastocyst). These results indicate that the choice of amino acids affects the competence of oocyte maturation and that Eagle's 20 AA with 0.2 mmol glutamine l(-1) is more efficient than the other amino acid formulations for maturation of rhesus monkey oocytes.

Nanobodies raised against monomeric α-synuclein distinguish between fibrils at different maturation stages.

Nanobodies are single-domain fragments of camelid antibodies that are emerging as versatile tools in biotechnology. We describe here the interactions of a specific nanobody, NbSyn87, with the monomeric and fibrillar forms of α-synuclein (αSyn), a 140-residue protein whose aggregation is associated with Parkinson's disease. We have characterized these interactions using a range of biophysical techniques, including nuclear magnetic resonance and circular dichroism spectroscopy, isothermal titration calorimetry and quartz crystal microbalance measurements. In addition, we have compared the results with those that we have reported previously for a different nanobody, NbSyn2, also raised against monomeric αSyn. This comparison indicates that NbSyn87 and NbSyn2 bind with nanomolar affinity to distinctive epitopes within the C-terminal domain of soluble αSyn, comprising approximately amino acids 118-131 and 137-140, respectively. The calorimetric and quartz crystal microbalance data indicate that the epitopes of both nanobodies are still accessible when αSyn converts into its fibrillar structure. The apparent affinities and other thermodynamic parameters defining the binding between the nanobody and the fibrils, however, vary significantly with the length of time that the process of fibril formation has been allowed to progress and with the conditions under which formation occurs, indicating that the environment of the C-terminal domain of αSyn changes as fibril assembly takes place. These results demonstrate that nanobodies are able to target forms of potentially pathogenic aggregates that differ from each other in relatively minor details of their structure, such as those associated with fibril maturation.

Nanobodies raised against monomeric α-synuclein distinguish between fibrils at different maturation stages

Nanobodies are single-domain fragments of camelid antibodies that are emerging as versatile tools in biotechnology. We describe here the interactions of a specific nanobody, NbSyn87, with the monomeric and fibrillar forms of α-synuclein (αSyn), a 140-residue protein whose aggregation is associated with Parkinson's disease. We have characterized these interactions using a range of biophysical techniques, including nuclear magnetic resonance and circular dichroism spectroscopy, isothermal titration calorimetry and quartz crystal microbalance measurements. In addition, we have compared the results with those that we have reported previously for a different nanobody, NbSyn2, also raised against monomeric αSyn. This comparison indicates that NbSyn87 and NbSyn2 bind with nanomolar affinity to distinctive epitopes within the C-terminal domain of soluble αSyn, comprising approximately amino acids 118-131 and 137-140, respectively. The calorimetric and quartz crystal microbalance data indicate that the epitopes of both nanobodies are still accessible when αSyn converts into its fibrillar structure. The apparent affinities and other thermodynamic parameters defining the binding between the nanobody and the fibrils, however, vary significantly with the length of time that the process of fibril formation has been allowed to progress and with the conditions under which formation occurs, indicating that the environment of the C-terminal domain of αSyn changes as fibril assembly takes place. These results demonstrate that nanobodies are able to target forms of potentially pathogenic aggregates that differ from each other in relatively minor details of their structure, such as those associated with fibril maturation. © 2013 Elsevier Ltd.

Population dynamics and maturation cycle of Camallanus cotti (Nematoda : Camallanidae) in the Chinese hooksnout carp Opsariichthys bidens (Osteichthyes : Cyprinidae) from a reservoir in China

The seasonal population dynamics and maturation cycle of the nematode Camallanus cotti in the posterior intestine of Chinese hooksnout carp Opsariichthys bidens have been studied in the Danjiangkou Reservoir of the Hubei Province in central China from September 2004 to November, 2005. The overall prevalence, mean abundance and intensity of C cotti among fish sampled (n = 700 fish) were 47%, 2.29 +/- 12.38 ( +/- S.D.) and 1-307 (average 4.89 +/- 17.74), respectively. The overall sexual ratio of female to male nematodes (excluding L3 and L4 juveniles) was 1.17:1. Statistical results showed weakly positive correlations betweerl fish length and the number of nematodes per host. The dynamics of infection of the nematode exhibited significant seasonal pattern in changes in mean abundance. A similar pattern was found for changes in nematode prevalence, although this was not statistically significant. Higher levels of infection were observed among fish sampled in summer months and the lower in the winter. Neither the prevalence nor the abundance of the parasite was significantly different between male and female hosts. The pattern of frequency distribution of the parasite in the host was found to be over-dispersed throughout the sampling period. In addition, studies on the development and maturation of the parasite in O. bidens revealed that development (maturation), recruitment of the next generation, and reproduction may be continuous year-round, although reproduction may peak during the winter. (c) 2007 Elsevier B.V. All rights reserved.