Identification of thiol group at the estrogen-binding site on the cytoplasmic receptor from rabbit uterus by affinity labeling

The interaction of estrogen with its receptor from rabbit uterus was studied using the affinity labeling technique. An affinity labeling reagent, 3-hydroxy-17β-(p-nitrophenyldithio)-1,3,5(10)-estratriene (Reagent A) was synthesized. The compound was designed to meet two requirements: (1) it binds specifically to the steroid-binding site of the receptor; (2) it has functional group capable of forming a covalent bond with an amino acid residue at or near the binding site. The first requirement was demonstrated by competitive binding assay and sedimentation analysis. If the binding affinity of 17β-estradiol is defined as 100, that of Reagent A is found to be 0.05. It was shown that the binding was specific for the estradiol receptor. The second requirement was examined by extraction of Reagent A with simple model compounds. 10 μM Reagent A gave rise to approx. 1.3 μM p-nitrothiophenol in the presence of a large excess of l-cysteine or reduced l-glutathione under the experimental conditions employed, indicating that a covalent bond was formed between Reagent A and the model compounds. Furthermore, I have presented evidence by using polyacrylamide gel electrophoresis that ³H-labeled Reagent A is linked covalently to the estrogen-binding site on the receptor of rabbit uterus. The experiment also fulfilled the first requirement. These results indicate that the thiol group present at the binding site is directly involved in the estradiol-receptor binding.     

Ontogenetic change in the relationship between metabolic rate and body mass in a sea bream Pagrus major (Temminck & Schlegel)

Ontogenetic changes in the relationship between resting rate of oxygen consumption and wet body mass were examined at 20° C with the sea bream Pagrus major ranging from 0.00020 g (weight just after hatching) to 270 g (weight at 530 days old). There was a triphasic relationship between oxygen consumption of an individual fish M (μl min⁻¹) and body mass W (g). During a very early stage (weight 0.00020–0.00025 g), corresponding to the pre-larval stage and with the transitional period to the post-larval stage, there was no substantial change in body mass. The mass–specific metabolic rate M/W (μl g⁻¹ min ¹) showed no clear relationship to body mass as expressed by the equation M/ W =4.86 + 1.47 D , where D is age in days. During the post-larval stage (weight 0.00031–0.005 g), M/W remained almost constant independent of body mass following the expression M = 12.5 W0 .949. During the juvenile and later stages (weight 0.005–270 g), M/ W decreased with increasing body mass following the expression M = 6.3 W ^0.821 which is significantly different from the expression for the post-larval stage ( P < 0.001). Ontogenetic changes in the metabolism-body mass relationship are discussed from the viewpoint of relative growth of organs with different metabolic activities.     

Latitudinal variation in blue tit and great tit nest characteristics indicates environmental adjustment

Aim The laying of eggs and the building of a nest structure to accommodate them are two of the defining characteristics of members of the class Aves. Nest structures vary considerably across avian taxa and for many species the structure of the completed nest can have important consequences both for parents and their offspring. While nest characteristics are expected to vary adaptively in response to environmental conditions, large‐scale spatial variation in nest characteristics has been largely overlooked. Here, we examine the effects of latitudinal variation in spring temperatures on nest characteristics, including insulatory properties, and reproductive success of blue tits, Cyanistes caeruleus, and great tits, Parus major. Location Great Britain. Methods Nests and reproductive data were collected from seven study sites, spread over 5° of latitude. The nest insulatory properties were then determined before the nests were separated into nest base material and cup lining material. Results As spring temperatures increased with decreasing latitude, the mass of the nest base material did not vary in either species, while the mass of the cup lining material and nest insulatory properties decreased in both species. This suggests that in response to increasing temperatures the breeding female reduces the mass of the cup lining material, thereby maintaining an appropriate microclimate for incubating and brooding. The mean first egg date of both species advanced with decreasing latitude and increasing spring temperatures, although clutch size and brood size at hatching and fledging did not vary. Main conclusions This is the first study to demonstrate that the nest‐construction behaviour of birds varies in response to large‐scale spatial variation in ambient temperatures. Therefore, nest composition reliably indicates environmental conditions and we suggest that studies of nest structure may be sentinels for the early signs of rapid climate change.     

Delayed egg production and a possible group effect in the blowfly Calliphora vicina

Abstract .Protein-fed Calliphora vicina , F₁ offspring of wild flies in two cages with lower and higher fly densities showed variable delay in starting oocyte vitellogenesis at ambient semi-natural temperatures in warm July–August weather in 1996 and 1997 at Durham in northern England (54°45' N). The high-density flies in 1996 showed no delay, in that the thermal sum (degree-days) experienced was 133, comparable to 18°C constant, assuming the lower threshold for egg maturation to be 5°C. Low-density cages and flies in a large outdoor cage (2 m³) in both years showed delays in production of first eggs of 34 days (thermal sum 293 degree-days) in 1996 and 32 days (396 degree-days) in 1997, and longer delays for other individuals. Delays in egg production at low densities relative to high densities seem to be a group effect of unknown mechanism.     

Electric field-induced orientation of l- and dl-phosphatidylcholine bilayers

Membrane orientation induced by an alternating electric field has been examined for the l-enantiomer and racemic dipalmitoylphosphatidylcholine (DPPC) bilayers. The orientation effect was measured by bending curvature of hairpin-like deformation of the multilamellar cylindrical tubes with varying field-strength, frequency and tube size. It has been observed that both l- and dl-DPPC tubes are similar in the profiles of field-strength dependence and frequency dependence on the curvature deformation, but different in the deformed curvatures. dl-DPPC tubes deform largely as compared with l-DPPC tubes. The square of the deformed curvature of dl-DPPC tubes is larger than that of l-DPPC by about 37% on average. The result indicates that the racemic membrane is responsive to the electric field as compared with the l-enantiomer membrane. This suggests that a hybrid arrangement of head groups of the racemic lipid leads an effective response of the membrane due to the head group orientation.     

Synthesis and radical-scavenging activity of C-methylated fisetin analogues

The radical-scavenging reaction of fisetin, a natural antioxidant found in strawberries, is known to proceed via hydrogen transfer to produce a fisetin radical intermediate. Thus, introduction of an electron-donating group into the fisetin molecule is expected to stabilize the radical, leading to enhanced radical-scavenging activity. In this study, fisetin derivatives in which methyl substituents were introduced at the ortho positions relative to the catechol hydroxyl groups were synthesized and their radical scavenging activities were evaluated and compared with that of the parent fisetin molecule. Among the methyl derivatives, 5′-methyl fisetin, in which the inherent planar structure of fisetin was retained, exhibited the strongest radical scavenging activity. Introduction of methyl substituents may be effective for the enhancement of various biological activities of antioxidants, particularly radical-scavenging activity.     

The amino acid sequence of the alpha subunit of mouse salivary androgen-binding protein (ABP), with a comparison to the partial sequence of the beta subunit and to other ligand-binding proteins

It has been suggested that three distinct genes,Abpa, Abpb, andAbpg, determine the three subunits of mouse salivary androgen-binding protein (ABP) (Dlouhy, S. R.,et al., Genetics 115, 535, 1987). We report the putative amino acid sequence of the subunit common to all forms of ABP, the Alpha subunit, and the partial amino acid sequence of the Beta subunit. These sequences have little in common, supporting the notion of at least two distinct genes coding for the subunits of the most common form of salivary ABP, the A:B dimer. A search of GenBank showed that these sequences have not been reported previously. The Beta subunit shows significant homology with helospectin, a member of the glucagon superfamily, but not enough homology to assign it to the family. No homology exists between ABP subunits and members of the ligand-binding carrier family of proteins nor does ABP show homology with other androgen-binding proteins. Particularly interesting is the observation that there is no relationship to rat prostatic steroid binding protein (PBP), given the similarities in protein tertiary structure, the numbers of subunits and their genes, and the earlier observation of ABP cross-reactive material in mouse prostate.Partial support for this work was provided by a PHS AREA award and by the Butler Academic Grants program. Both sources of support are greatly appreciated.A portion of this work constituted partial fulfillment of the honors thesis requirement for Butler University.     

Effect of Hyper-Osmotic Stress On Alanine Content of Leishmania Major Promastigotes

ABSTRACT Earlier studies showed that Leishmania major promastigotes are sensitive to osmotic conditions. A reduction in osmolality caused the cells to shorten and to rapidly release most of their large internal pool of alanine. In this study some effects of hyper-osmotic stress were examined. an increase in osmolality of the culture medium from 308 to 625 mOsm/kg caused only a small decrease in growth rate. When cells grown in the usual culture medium (308 mOsm/kg) were washed, resuspended in iso-osmotic buffer, and subjected to acute hyper-osmotic stress by addition of mannitol, the alanine content increased even in the absence of exogenous substrate. Promastigotes, depleted of alanine by a 5-min exposure to hypo-osmotic conditions, also synthesized alanine when resuspended in iso-osmotic buffer. Washed cells resuspended in iso-osmotic buffer consume their internal pool of alanine under aerobic conditions, Rates of consumption decreased on addition of mannitol, becoming zero at about 440 mOsm/kg. At higher osmolalities, alanine synthesis occurred. to estimate whether proteolysis could account for alanine synthesis in the absence of exogenous substrate, cells that had been grown with [1-¹⁴C]leucine were washed and resuspended under hypo-, iso-, and hyper-osmotic conditions and the amounts of ¹⁴CO₂ and ¹⁴C-labelled peptides released in 1 h were measured. Little proteolysis occurred under these conditions, but the possibility that proteolysis was the source of the alanine increase, observed in response to hyper-osmotic stress, cannot be ruled out.     

长江安庆新洲水域鱼类群落结构及多样性

沙洲水域环境良好,饵料资源丰富,栖息生境多样,为鱼类的生长繁殖提供了优良的生存环境。为了解长江安庆新洲水域鱼类群落结构特征,于2017年4月、7月、10月和12月对安庆江段新洲水域鱼类群落进行季节性调查。共采集鱼类64种,分属5目11科48属,其中62.5%为鲤科鱼类。以物种数和多样性指数分析群落多样性特征,结果表明新洲水域鱼类种类多样性水平较高。单因素方差分析表明,该群落多样性季节差异显著(P0.05),空间差异不明显。新洲水域鱼类群落优势种为鳊(Parabramis pekinensis Basilewsky, 1855)、鲤(Cyprinus carpio Linnaeus, 1758)、贝氏■(Hemiculter bleekeri Warpachowsky, 1887)、银鮈(Squalidus argentatus Sauvage et Dabry, 1874)和似鳊(Pseudobrama simoni Bleeker, 1864)。4种摄食功能群中,杂食性(42.19%)和肉食性(35.94%)鱼类物种数比例较高;3种生态类群中,淡水定居性鱼类占绝对优势(84.37%);3种栖息水层类型中,底层鱼类物种数比例较高,占37.50%。大型经济鱼类占总渔获物比例低(0.01%),但个体较大,因而相对重要性指数(IRI)高。总体上,新洲鱼类群落多样性和丰富度指数较高,均匀度指数偏低,个体小型化趋势明显。捕捞强度过大、水利工程建设导致的江湖阻隔及外来物种入侵是新洲水域渔业资源衰退的主要因素。由此,建议持续开展长江渔业资源监测,加强长江干流沙洲水域渔业资源保护。