Metal complexes as artificial proteases: toward catalytic drugs

Catalytic cleavage of the backbone of a protein related to a disease may cure the disease. Owing to the catalytic nature of the protein inactivation, the drug dosage and the side effects can be reduced with the catalytic drugs. Catalytic drugs can be designed even for proteins lacking active sites. Effective artificial proteases have been designed for proteins or oligomers of oligopeptides. The Co(III) complex of cyclen has been used as the catalytic center for peptide hydrolysis. Binding sites of the catalysts that recognize the targets have been searched by using various kinds of chemical libraries. Some of the artificial metalloproteases reported till date offer a new therapeutic option for amyloidoses (e.g. Alzheimer's disease, type 2 diabetes mellitus, Parkinson's disease, mad cow's disease, etc.).     

Recruitment of Antarctic marine eukaryotes onto artificial surfaces

Activities related to Antarctic research stations have caused significant local impacts on the marine environment, potentially affecting the recruitment of benthic invertebrates. Herein, we report the community structure of recruiting marine eukaryotes onto artificial substrata using molecular techniques. Slides were deployed at three sites adjacent to McMurdo Station, Scott Base, and Cape Armitage in McMurdo Sound. Denaturing gradient gel electrophoresis (DGGE) analysis revealed complex and diverse eukaryotic communities had established on artificial surfaces deployed at a range of site and depth regimes after 12 months. Analysis of similarity results detected significantly greater variability in community profiles among sites than within sites. The nonmetric multidimensional scaling plot constructed from DGGE banding patterns revealed different benthic communities had established at 12 and 18 m depths. Despite this, the variation in community composition was greater among sites than between depths, especially at Cape Armitage and Scott Base. Sequence analysis of excised DGGE bands revealed a predominance of arthropod and dinoflagellate sequences at Cape Armitage. In contrast, a wide diversity of phyla including cnidaria, bryozoa, protozoa, dinoflagellates, arthropods, platyhelminths, and annelids were present adjacent to the two research stations. The abundance of diatoms detected in Cape Armitage benthic assemblages exceeded the abundance of diatoms from McMurdo Station and Scott Base by almost two orders of magnitude. The discovery that distinct eukaryotic communities recruit at different sites and depths is probably due to complex interactions between multiple factors including water quality, larval supply, and light. The detection of sessile phyla on slides at each of the sites indicates that the pollution profiles present at each site is not an impediment to successful recruitment of these species.     

Toward protein-cleaving catalytic drugs: artificial protease selective for myoglobin

A protein-cleaving catalyst highly selective for a disease-related protein can be used as a catalytic drug. As the first protein-cleaving catalyst selective for a protein substrate, a catalyst for myoglobin (Mb) was designed by attaching the Cu(II) or Co(III) complex of cyclen to a binding site searched by a combinatorial method using peptide nucleic acid monomers as building units. Various linkers were inserted between the catalytic Co(III) center and the binding site of the Mb-cleaving catalyst. Kinetic data revealed catalytic turnover of the Mb cleavage by the Cu(II) or Co(III) complex. MALDI-TOF MS revealed cleavage of the polypeptide backbone of Mb at selected positions. N-Terminal sequencing of the cleavage products identified the cleavage site and provided evidence for the hydrolytic nature of the Mb cleavage. Various chelating ligands were tested as the ligand for the Co(III) center of the Mb-cleaving catalyst. Among the nine chelating ligands examined, only cyclen and its triaza-monooxo analogue manifested catalytic activity.     

Nonfouling behavior of polycarboxybetaine-grafted surfaces: structural and environmental effects

Zwitterionic carboxybetaine (CB) has unique dual functionality for ligand immobilization on a nonfouling background. The properties of CB groups depend on their spacer groups between the positive quaternary amine groups and the negative carboxyl groups and environmental factors (e.g., ionic strengths and pH values). In this work, five polycarboxybetaines were prepared, including one polycarboxybetaine methacrylate (polyCBMA) and four polycarboxybetaine acrylamides (polyCBAAs) with different spacer groups. The polymers were grafted from a gold surface covered with initiators using surface-initiated atom transfer radical polymerization. Fibrinogen adsorption was measured as a function of ionic strengths and pH values using surface plasmon resonance sensors. The responsive protein adsorption on four polyCBAAs was mapped out. Results show that most of these surfaces exhibit high protein resistance in a wide range of ionic strengths and are more effective than zwitterionic self-assembled monolayers. Although protein adsorption tends to increase at low ionic strength and low pH value, it is still very low for polycarboxybetaines with a methylene, an ethylene, or a propylene spacer group but is more evident for polyCBAA with a longer spacer group (i.e., a pentene group). The response to ionic strengths and pH values can be attributed to the antipolyelectrolyte and protonation/deprotonation properties of polycarboxybetaines, respectively. Both of these properties are related to the spacer groups of CBs.     

Platelets and the immune continuum

Platelets are anucleate cells that are crucial mediators of haemostasis. Most immunologists probably don't think about platelets every day, and may even consider these cells to be 'nuisances' in certain in vitro studies. However, it is becoming increasingly clear that platelets have inflammatory functions and can influence both innate and adaptive immune responses. Here, we discuss the mechanisms by which platelets contribute to immunity: these small cells are more immunologically savvy than we once thought.     

GABA and the behavioral effects of anxiolytic drugs

Much recent research has shown that benzodiazepine binding sites in the central nervous system are associated with GABA receptors. It is therefore possible that the pharmacological and therapeutic effects of benzodiazepines and drugs with similar profiles are mediated through GABAergic mechanisms. In this paper the evidence is considered for a possible involvement of GABA in the behavioral effects of anxiolytic drugs. There are a number of reports that the behavioral actions of anxiolytics can be antagonised by GABA antagonists such as bicuculline or picrotoxin but there are many contradictory findings and these drugs are difficult to use effectively in behavioral studies. In general, GABA agonists do not exert anxiolytic-like behavioral effects after systemic injection but intracerebral administration of muscimol has been shown to produce benzodiazepine-like actions. Although a number of questions remain unanswered, current evidence does not provide strong support for a role for GABA in the behavioral effects of anxiolytic drugs.     

激光扫描共聚焦显微镜术检测单个血小板钙波动方法的探讨

目的 探讨血小板贴壁的简易方法,同时,观察单个血小板激活前后细胞内钙波动及形态变化的规律。方法 采用多种粘附剂固定血小板,利用钙荧光探针（Fluo-3/AM)（终浓度10－20μmol)进行染色,在激光扫描共聚焦显同镜检测下加入二磷酸腺苷,观察和分析血小板内Ca^2 浓度及形态变化。结果 多聚赖氨酸促进血小板贴壁固定的效果最佳;单个血小板活后细胞内Ca^2 浓度为激活前的128％,形态圆形或椭形转为不规则形,有空泡,突起形成。结论 多聚赖氨酸是血小板贴壁的理想粘附剂,本方法能简易,快捷地监测血小板激活过程中胞浆内钙离子动态变化及形态改变,有助于血小板功能的深入研究。     

Oscillatory potentials of the cat retina: effects of adrenergic drugs

When reserpine was used as a monoaminergic depletion agent, a diminution or suppression of certain components of the oscillatory potentials of the electroretinogram of the cat was observed. Administration of L-Dopa to previously reserpinized cats restored the altered OP components. The possible interference of systematic disturbances provoked by the drugs was studied by means of artificial variations of blood pressure. Results reported in this paper suggest that reserpine and L-Dopa are acting on catecholamine-dependent retinal structures, probably the dopamine-containing population of amacrine cells. An hypothesis is advanced concerning the possible role of tangential structures in the generation of the oscillatory potentials of the retina.     

Euphorigenic drugs: effects on the reward pathways of the brain.

This report summarizes a number of experiments designed to examine the changes in the threshold for intracranial self-stimulation (ICSS) in the rat after the administration of morphine and a number of narcotic agonist-antagonists, as well as three nonnarcotic drugs that have extensive nonmedical use (cocaine, d-amphetamine, and phencyclidine). The results of these experiments clearly indicate that morphine lowers the threshold for ICSS and, furthermore, there appears to be little or no tolerance to this effect. The only mixed agonist-antagonist that consistently lowered the ICSS threshold was pentazocine. Cocaine, d-amphetamine, and to a lesser degree, phencyclidine also lowered the ICSS threshold. These results suggest that the abuse liability of these agents may be directly related to their ability to sensitize the neural substrate involved with natural reward.     

Respiratory effects of the Jarvik-7 artificial heart

In five anesthetized patients with a Jarvik-7 artificial heart, pulmonary volume displacements generated by cardiogenic oscillations were measured using an indirect spirometric method. Consequences on gas exchange were also evaluated during a 15-min period of apnea by use of a tracheal insufflation of pure O2 at a constant flow rate of 20 l/min. The Jarvik-7 artificial heart generated a mean pulmonary volume displacement of 105 +/- 29 (SD) ml/heart beat. After 15 min of apnea, arterial PCO2 (PaCO2) significantly increased from 29 +/- 5 to 47 +/- 6 (SD) Torr. PaCO2 increased by 0.8 Torr/min from the 5th to the 15th min of apnea. Mean arterial PO2, mean pulmonary shunt, mean O2 consumption, and mean metabolic production of CO2 did not change significantly during the apnea period. Because cardiac output was kept constant during the study, O2 transport was adequately maintained throughout the apnea period. In patient 1, where the period of apnea was continued for 60 min, PaCO2 progressively increased until the 45th min and then remained stable at 61 Torr during the last 15 min of apnea. This "plateau" corresponded to an alveolar ventilation of 3,907 ml/min, representing 69% of the alveolar ventilation calculated during conventional mechanical ventilation. In conclusion, the Jarvik-7 artificial heart provides a potent respiratory support through the cardiogenic oscillations it generates.