IT has been reported1 that washed human platelets incubated with 14C-labelled cytidine monophpsphate-N-acetyl sialic acid (CMP-NANA) in the presence of homogenized rat liver as the source of sialyltransferase showed an increase in the amount of sialic acid bound to the platelet membrane: illustration
DURING multiple passages, fibroblast cell lines retain the chromosome number and genetic defects of the original donor1,2. Here we show, by comparing the modes of glycolysis in foetal and non-foetal skin cultures, that-fibroblasts retain the expression of their original developmental state. illustration
CARBONIC anhydrase catalyses hydration of aldehydes1–3: illustration I have studied the relevance of hydration of glyceraldehyde-3-phosphate (GAP) in its oxidation by GAP dehydrogenase. GAP dehydrogenase activity was assayed spectrophotometrically by measuring the increase in absorbance at 340 nM of a reaction mixture containing 0.1 M Tris-HCl (pH 8.5), 17 mM sodium arsenate, 5 mM cysteine, 20 mM NaF, 1 mM NAD, 0.1 U/ml. of GAP dehydrogenase (Sigma) and 4 µM GAP.
REDUCED glutathione (GHS) is maintained in substantial concentration (3 × 10?3 M) within neurones1. The availability of new thiol-oxidizing agents like diamide2,3, which freely enter cells and the principal action of which is to reduce the GSH concentration by conversion to GSSG (equation 1), promoted an investigation of the effects of these agents on release of transmitter at frog myoneural junctions. illustration
IT has been shown that rabbit anti-serum against rat serum (RARS) injected intravenously into rats produces fatal anaphylactic shock1. This was interpreted as a reaction of RARS with γ-globulins adsorbed on the cell surface2. It therefore seemed reasonable to investigate the same effect on enhancing antibodies by injecting RARS into rats bearing Yoshida ascites sarcoma (YAS). The result was as expected–delayed death. It has been suggested that the “de-enhanced” tumour cells become more susceptible to lymphocytes3. Although the presence of enhancing antibodies coating YAS cells has not actually been shown, if they are responsible for the observed phenomena, other agents acting on γ-globulins should result in both anaphylaxis and “de-enhancement”. We therefore used cathepsins isolated from white blood cells, which act specifically on β and γ-globulins4. illustration
THE molecular mechanism for initiating DNA replication can be studied using a subcellular system. Rao and Johnson1 found that HeLa cells in the pre-DNA-synthetic (G-1) period of the cell cycle initiate DNA synthesis after fusion with cells that are in the DNA synthetic (S) period. A previous subcellular system of DNA replication from HeLa cells2–4 consisted of intact nuclei, supplemented with the four deoxy-nucleoside triphosphates, salt, ATP and a cytosol factor. The nuclei in this system appeared to be permeable to proteins and DNA synthesis was very similar to that within intact cells. We report here the initiation of DNA synthesis in nuclei isolated from HeLa cells. Our results suggest that, with the synchronization method used, a small percentage of dormant G-1 nuclei can be stimulated by S-phase cytoplasm; this would be the case if the cells were receptive to stimulation for only 30–60 min during the cell cycle. illustration
USEFUL antisera specific for thymocytes and peripheral T lymphocytes have only been widely available in the form of antisera to thymic isoantigens of the mouse1. We describe here the preparation and properties of a heterologous antiserum to guinea-pig thymocytes which is rendered specific for T lymphocytes after absorption with a pure population of B lymphocytes. We have already described2,3 the properties of the transplantable acute lymphatic leukaemia L2C of inbred strain two guinea-pigs. The L2C leukaemia cell is characterized as a B cell by the presence of surface immunoglobulin of the λ2 class, the secretion of a small amount of λ2 immunoglobulin and the presence of a receptor for antigen-antibody-complement (C3) complexes characteristic of the B cell population4. Because, as will be shown, the antiserum is specific both for thymocytes and thymus derived lymphocytes, it will be referred to as anti-thymus derived cell (anti-TDC) serum. The availability of such an antiserum for a species in which the in vivo and in vitro manifestations of delayed hyper-sensitivity are so easily demonstrated may prove to be highly advantageous. illustration
THE ribonuclease, barnase, produced by Bacillus amyloliquefaciens has a molecular weight of 12,382, consisting of 110 amino-acid residues. It is one of the smallest proteins containing neither disulphide bonds nor non-peptide cross-Bnks which nevertheless maintain a well defined tertiary structure1,2. The next smallest reported enzyme of similar nature is the lysozyme of phage T4, with 160 residues. The barnase structure is reversibly destroyed by denaturing solvents or heat2, in what approximates a one step, highly cooperative, transition. Studies of this reaction should be very useful in illustration approaching the general problem of sequence-determined folding in proteins. In particular, thermodynamically meaningful quantitative differences in the stability of various genetic variants and chemically modified, or synthetic, barnases could be measured. Some work has been reported on the effect of various environmental parameters on the transition3 as well as the effects of modification by carboxypeptidases4. Full utilization of such data requires knowledge of both amino-acid sequence and three dimensional structurs. The complete amino-acid sequence is reported here (Fig. 1). The sequence was obtained by conventional procedures involving analysis of peptides isolated after hydrolysis of the native or modified protein by various proteases.
THE carboxy-terminal tetrapeptide of gastrin, Trp-Met-Asp-Phe-NH2, has the same biological activity as the parent hormone1. Morley2,3 showed that certain substitutions in the Trp, Met and Phe positions gave active analogues and concluded that these positions are concerned only with binding at the site of action. In contrast, the only substitution in aspartic acid that gave activity was tetrazolyl for the β-carboxyl, thereby maintaining a proton donor of similar size at this position. Morley concluded that the aspartyl residue has a functional rather than a binding role and presumably is indispensable. The carboxy-terminal octapeptide of cholecystokinin (OP-CCK) contains the carboxy-terminal tetrapeptide of gastrin and has gastrin-like activity4. We report here that the synthetic analogue of OP-CCK, in which alanyl is substituted for aspartyl at the position in question illustration, stimulates gastric acid secretion. In conscious cats with gastric fistulas5, dose-response curves were established for gastric acid response to OP-CCK, 7-Ala OP-CCK and pentagastrin6. The peptides were given by continuous intravenous infusion and response is taken as peak 10 min output during a 30 min infusion at each dose level (Fig. 1). Assigning a potency of 1 to OP-CCK the relative molar potency of 7-Ala OP-CCK is about 1/110 and of pentagastrin is about 1/4. We also tested the analogue of gastrin tetrapeptide in which alanyl is substituted for aspartyl illustration and found no detectable stimulation of acid secretion at doses as high as 15 mg/kg h, confirming similar negative findings by Morley2 in rat. This suggests that the weak action of alanyl substituted analogues cannot readily be detected without the enhancement of potency conferred by the sulphated tyrosyl of OP-CCK7. Ondetti et al.7 showed that 7-Ala OP-CCK contracts guinea-pig gallbladder with a potency about 1/150th that of OP-CCK, comparable with that reported here for acid secretion. This suggests that the same part or parts of the molecule are required for cholecystokinetic action and for gastric secretory action; the aspartyl residue in the penultimate position is dispensable for both of these actions. On the assumption that gastrin and CCK act at the same site8, we propose that the corresponding aspartyl residue of gastrin is similarly dispensable. For a direct test of this hypothesis, studies are needed of the synthetic analogue in which alanyl is substituted for the penultimate aspartyl in gastrin hepta-decapeptide, perferably gastrin II with sulphated tyrosyl because it is more potent than gastrin I in certain species9.
The 1A1 ground and the first 1B2 excited states of the methylenecyclopropene (triafulvene) are described by localized wave functions, based on 20 structures valence bond structures. The results are compared to CASSCF(4,4) calculations for both the energetics and the dipole moment. Additional calculations with partial electronic delocalization are presented, and it is shown that the dipole moment modification does not correspond to a situation where the antiaromatic situation prevails (with 4n electrons in the cycle). Part of the analysis uses a “trust factor” that helps to decide if a wave function is appropriate to describe a given state. The trust factor compares the VB wave function to the CASSCF’s with their overlap. Finally, the valence bond density is used to produce density maps that illustrate the electron transfer upon excitation.
Graphical Abstract A projector-based method compares CASSCF wave functions to local wave functions, including Lewis structures as shown in the picture. A “trust factor” (τ) is obtained. Both the ground state and the first excited state of the methylenecyclopropene are discussed
A perfectly planar Al13+ cluster (CI) and a quasi-planar Al13+ cluster (CII) have been found for the first time. Both clusters have a triangular core surrounded by a set of ten Al atoms in the form of a ring. These cationic clusters have substantial aromatic character. The planar CI cluster has local antiaromatic patches within global aromatic sea. It is doubly aromatic having both σ and π aromatic character. The quasi-planar CII cluster is also aromatic but it has more σ-delocalization.
The present study reports the geometries, electronic structures, growth behavior, and stabilities of neutral and ionized copper-doped germanium clusters containing 1–20 Ge atoms within the framework of linear combination of atomic orbitals density functional theory (DFT) under the spin-polarized generalized gradient approximation. It was found that Cu-capped Gen (or Cu-substituted Gen+1) and Cu-encapsulated Gen clusters mostly occur in the ground state at a particular cluster size (n). In order to explain the relative stabilities of the ground-state clusters, parameters such as the average binding energy per atom (BE), the embedding energy (EE), and the fragmentation energy (FE) of the clusters were calculated, and the resulting values are discussed. To explain the chemical stabilities of the clusters, parameters such as the energy gap between the highest occupied and the lowest unoccupied molecular orbitals (the HOMO–LUMO gap), the ionization energy (IP), the electron affinity (EA), the chemical potential (μ), the chemical hardness (η), and the polarizability were calculated, and the resulting values are also discussed. Natural atomic orbital (NAO) and natural bond orbital (NBO) analyses were also used to determine the electron-counting rule that should be applied to the most stable Ge10Cu cluster. Finally, the relevance of the calculated results to the design of Ge-based superatoms is discussed.
Figure Contributions of the valance orbitals of the Ge and Cu atom(s) to the HOMO of the ground-state icosahedral Ge10Cu cluster obtained from NBO analysis. The numbers below the clusters represent the occupancies of the HOMO orbitals
Using density functional theory (DFT) and molecular dynamics (MD), we studied the interaction of a titanium atom with a half of a C60 fullerene (i.e., C30), formed from the corannulene structure with a pentagonal base. We considered atmospheric pressure and 300 K. We found that the most stable adsorption of the titanium atom on C30 occurs in the concave surface of the molecule. Afterward, we investigated the interaction of the system C30-titanium with carbon monoxide and carbon dioxide molecules, respectively. We found that each of these molecules is chemisorbed, with no dissociation. The value of the adsorption energy for the carbon monoxide molecule varies from ?0.897 to ?1.673 eV, and for the carbon dioxide molecule, it is between ?1.065 and ?1.274 eV. These values depend on the initial orientation of these molecules with respect to TiC30.
The aldol reaction in the presence of L-proline acting as an organocatalyst is a well-known example of asymmetric synthesis. Many theoretical and experimental studies have been carried out to probe the mechanism of this reaction. In this work, two levels of density functional theory in the gas phase and DMSO were used to elucidate the best pathways for this reaction, with the enamine and enol considered intermediates and L-proline considered either a reactant or a facilitator. The calculations indicated that both intermediates are formed simultaneously in the reaction medium. Interestingly, the formation of the enamine intermediate predominates in DMSO at room temperature, whereas the enol becomes the predominant intermediate upon the addition of water.
A topological analysis based on density functional electronic and spin densities of the bonding characteristics in a series of Fe, Ru, Os, Tc and Rh dimers and trimers bridged, respectively, by μ-1,8-naphthyridine (nap) and μ-2,2′-dipyridylamine (dpa) is presented. By this simple qualitative analysis, we were able to determine the electronic ground state and correlated bonding order for a number of complexes potentially involved in extended metal atom chains (EMAC). Furthermore, we showed in the Ru dimer that it was possible to control the spin state simply by changing the bonded counter-anion.
Phospholipase A2 (PLA2) is one of the key enzymes involved in the formation of inflammatory mediators. Inhibition of PLA2 is considered to be one of the efficient methods to control inflammation. In silico docking studies of 160 selected indole derivatives performed against porcine pancreatic PLA2 (ppsPLA2) suggested that, CID2324681, CID8617 (indolebutyric acid or IBA), CID22097771 and CID802 (indoleacetic acid or IAA) exhibited highest binding energies. In silico analysis was carried out to predict some of the ADME properties. The binding potential of these compounds with human non pancreatic secretory PLA2 (hnpsPLA2) was determined using molecular docking studies. In order to corroborate the in silico results, enzyme kinetics and isothermal titration calorimetric analysis of the two selected compounds, IAA and IBA were performed against ppsPLA2. From the analysis, it was concluded that IAA and IBA can act as competitive inhibitors to the enzyme and may be used as anti inflammatory agents.
The serine/threonine protein phosphatase type 5 (PP5) is a promising target for designing new antitumor drugs. This enzyme is a member of the PPP phosphatases gene family, which catalyzes a dephosphorylation reaction: a regulatory process in the signal transduction pathway that controls various biological processes. The aim of this work is to study and compare the inhibition of PP5 by ten cantharidin-like inhibitors in order to bring about contributions relevant to the better comprehension of their inhibitory activity. In this theoretical investigation, we used molecular dynamics techniques to understand the role of key interactions that occur in the protein active site; QM calculations were employed to study the interaction mode of these inhibitors in the enzyme. In addition, atoms in molecules (AIM) calculations were carried out to characterize the chemical bonds among the atoms involved and investigate the orbital interactions with their respective energy values. The obtained results suggest that the Arg275, Asn303, His304, His352, Arg400, His427, Glu428, Val429, Tyr451, and Phe446 residues favorably contribute to the interactions between inhibitors and PP5. However, the Asp271 and Asp244 amino acid residues do not favor such interactions for some inhibitors. Through the QM calculations, we can suggest that the reactional energy of the coordination mechanism of these inhibitors in the PP5 active site is quite important and is responsible for the inhibitory activity. The AIM technique employed in this work was essential to get a better comprehension of the transition states acquired from the mechanism simulation. This work offers insights of how cantharidin-like inhibitors interact with human PP5, potentially allowing the design of more specific and even less cytotoxic drugs for cancer treatments.
