A spectroscopic study of metal ion and ligand binding to β-lactamase II

β-Lactamase II has two metal-binding sites. The electronic spectra of Cd(II)- and Co(II)-substituted β-lactamase II have been investigated. It is suggested that a thiol ligand is involved in metal binding at the first site. The stoichiometric dissociation constants for Co(II) binding to β-lactamase II were estimated to be 0.13 and 2.66 mM (pH 6.0, 4°C, 1 M NaCl) by equilibrium dialysis. Competition between Zn(II) and Co(II) for the first metal binding site suggests a value of 0.7 μM (pH 6.0, 30°C, 1 M NaCl) for the dissociation constant o Zn(II).The electronic spectra of the Co(II) enzyme lead to the suggestion that the coordination geometries around the metal ions in the first and second sites are related to those of a distorted tetrahedron and octahedron, respectively.     

Inhibition of co-operative enzymes by substrate-analogues: Possible implications for the physiological significance of negative co-operativity illustrated by phenylalanine metabolism in higher plants

The “Hill” equation for co-operative binding-systems has been extended to describe the effect of substrate-analogue on the binding of substrate to an oligomeric protein. It is demonstrated that the more negatively co-operative the binding-system, the more sensitive is the binding of substrate to inhibition by increases in the relative concentration of substrate-analogue. It is proposed that the physiological significance of negative co-operativity for enzymes may be complementary to the physiological significance of positive co-operativity. The effect of negative co-operativity is to make substrate binding more sensitive to inhibition by relative increases in the concentration of substrate-analogue (e.g. for many enzymes product of the reaction) at the expense of decreased sensitivity of substrate binding to relative changes in substrate concentration compared to a system with equivalent, independent substrate binding sites. In contrast, the effect of positive co-operativity is to make the enzyme more sensitive to relative changes in substrate concentration at the expense of decreased sensitivity to inhibition by relative increases in product concentration, compared to an enzyme without co-operative binding.     

The biology of the stoat (Mustela erminea) in the National Parks of New Zealand III. Morphometric variation in relation to growth,geographical distribution,and colonisation

Abstract

A total of 1599 stoats were collected from 14 study areas (including all 10 National Parks) from 1972 to 1976. Samples were larger in summer, and contained more females. Young stoats are born in September-October, and females reach adult weight by the following March, though males not until after August. There was significant geographic variation in the body size of adult stoats sampled: males from lowland podocarp/broadleaved forests averaged 3% smaller than males from upland beech forests in skull length, and 4% smaller in head-and-body length. This pattern was repeated, less clearly, in females and in young (approximately 2–5 months old). In contrast with stoats in Britain, assumed to be still the same size as the colonising stock introduced into New Zealand in 1884 and subsequently, males from lowland podocarp forests were unchanged or possibly smaller, and males from upland beech forests were larger; females were larger in all habitats. In males, the extent of geographic variation is almost as great in New Zealand as in the whole of continental Europe. Possible explanations of this pattern are discussed.     

Metal ion-biomolecule interactions. II. Methylmercuration of the deprotonated amino groups in adenine,guanine, and cytosine derivatives,and its relationship to amino group acidity

Proton nmr spectroscopic evidence is presented for methylmercury(II) binding to the deprotonated amino groups in adenosine, 9-methyladenine, guanosine, 1-methylguanosine, and cytidine under basic conditions. Except for the guanosine case, ¹H nmr spectra of the products from aqueous or ethanolic 1:1 mixtures of substrate and MeHgOH are consistent with methylmercuration of the deprotonated amino groups. Guanosine undergoes initial binding of MeHg to N₁, and a second equivalent of MeHgOH is necessary to effect amino binding. The nmr spectra of the complexed adenine derivatives suggest that different geometrical isomers exist in (CD₃)₂SO solution, reflecting the partial double bond character of the C₆N bond in these systems. Using a correlation relating the magnitude of the ¹⁹⁹Hg-¹H coupling constant (J) for MeHg-ligand complexes with the ligand pK_a (J = ?3.88 pK_a + 248.5, extending over 13 pK units, based on a variety of N and O donor ligands), estimates (± 0.3 pK unit) of the pK_as of the amino groups of the above substrates have been made. In this way, pK_a values of 15.5 (cytidine), 17.0 (adenosine and 9-methyladenine), 15.1 (guanosine), and 14.9 (1-methylguanosine) are obtained. In the cases where comparisons with literature pK_a data can be made, good agreement is found.     

诊间自动预约模式在超声检查预约中的应用

目的 描述和分析诊间自动预约模式对超声检查预约的影响。 方法 在诊间自动预约应用前后,分别以187名和160名患者为调查对象,采用两独立样本数据的t检验对数据进行对比。结果 81.0%的超声检查患者减少了1次排队,预约排队等候时间缩短了23.6分钟（P＜0.05）,检查等候时间缩短了7.0分钟（P＜0.05）。结论 诊间自动预约模式优化了超声检查预约服务,缩短了患者的排队等候时间,可推广至其他医技检查。     

Diversity of macrofungi and exploitation of edible mushroom resources in the National Park “Appennino Lucano,Val D'Agri,Lagonegrese” (Italy)

An investigation on the macrofungal diversity of the National Park “Appennino Lucano, Val D'Agri, Lagonegrese” (Basilicata, southern Italy) was carried out, together with an evaluation of wild edible mushrooms in agro-forest ecosystems and their possible exploitation as a new source of food and revenue in rural and under-developed areas of this region. An unpublished list of 249 mushroom taxa (229 Basidiomycota and 20 Ascomycota), mostly belonging to the genera Tricholoma, Tuber, Russula, Amanita, Boletus, Lactarius, Mycena, Agaricus, Clitocybe and Hygrophorus, growing in this important natural area is provided together with ecological characterization of the recorded specimens. Inonotus obliquus is a new record for Basilicata, while Mycena bresadolana is reported for the first time from southern Italy. The presence of Amaurodon mustialaënsis, an extremely rare species of the northern hemisphere reported for the first time on white fir, is noteworthy. Among the previous findings, 116 species of edible mushrooms are presented and discussed in view of their relative abundance and edibility.     

Nuclear magnetic resonance studies of the phenylalanine residues of eukaryotic cytochrome c

The resonances of Phe 82 and Phe 10 in the nuclear magnetic resonance spectra of horse cytochrome c are reassigned using nuclear Overhauser enhancements. The reassignments provide new information about the oxidation state linked conformation change of cytochrome c. The region of the protein now known to be affected by the change extends to the part of the protein close to Phe 10.     

Association Constants for Metal Hexacyanide Binding to Cytochrome c

The binding of[Co(CN)₆]^3?, and that of[Fe(CN)₆]^3? and [Ru(CN)₆]^4? using a competitive method, to horse cytochrome c has been studied by ⁵⁹ Co NMR spectroscopy. At I = 0.07 M, without added salt and in ²H₂O at ph* 7.3 (measured in ²H₂O) and 25°C, there are at least two binding sites on ferricytochrome c and ferrocytochrome c for [Co(CN)₆]^3?. Association constants were determined to be 2.0 ± 0.6 × 10³M^?1 and 1.5 ± 0.5 × 10²M^?1 respectively. with no effect of the oxidation state of the cytochrome. At higher ionic strength (I = 0.12 M adjusted with KCl the binding markedly decreased, and, although it was not possible to determine the precise binding stoichiometry and magnitude of association constants, it is clear that the association constants are ≤ 1.5 × 10^tM^?1 The binding of [Ru(CN)₆]^4? at I = 0.07, without added salt and in ²H₂O at pH 1.3 and 23°C, was not precisely defined, but its binding strength relative to that of [Fe(CN)₆]^3? was determined. Extrapolating this to I = 0.12 (KCl) suggests that under these conditions the association constant for [Ru(CN)₆]^4? binding to ferricytochrome c is ≤ 3 × 10²M^?1.     

Strengthening the Natural and National Park system of Iberia to conserve vascular plants

The diversity of the Iberian vascular flora has been investigated using WORLDMAP versions 3.08 and 3.18. Two data sets scoring plant distributions as presences within the Iberian Peninsula were compiled; one for 2133 species at 50 × 50 km grid and the other for 801 species at 10 × 10 km map grids. Patterns of biodiversity were determined using the diversity measures of species richness, range-size rarity and character richness diversity. Using the diversity measures, combined with an area selection method, maps of priority areas were calculated using iterative procedures. Near minimum sets (NMSs) for both scales were calculated. Comparison of the NMS for the 10 × 10 km grid with the near minimum set for existing reserves (NMSER) showed that at least 2% more of the land surface would be required above and beyond the existing protected area network, currently comprising 6% of the area, to ensure representation of all species at least once as listed within the present data-base. It is demonstrated that reserve systems selected on a variety of different criteria are suboptimal when compared to particular groups of target organisms with a definite goal of representation for conservation. Calculating efficiency of existing reserve systems and accounting for all taxa identifies precisely the extra required areas for the protected area system to satisfy particular goals of representation.