Mechanism of Nitrite-Induced Germination of Clostridium perfringens Spores

A study has been undertaken to understand the mechanism(s) of the nitrite-induced germination of Clostridium perfringens S40 spores. An increase in germination rates of the spores in response to increasing NaNO2 concentrations was entirely dependent on both pH and temperature of incubation. Low pH and high temperature were effective in accelerating the germination rate, the maximal germination level being reached at pH 4.0 and 60°C in the presence of 0.5 M NaNO₂. On the basis of germination rate, the activation energy (μ) for the nitrite-induced germination calculated was approximately 9.9 kcal/mol. Germination was greatly stimulated after pretreatment of spores with DTT at pH 10.5 to remove the coats. Furthermore, cortical fragments prepared from spores of the same organism were lysed not only by lysozyme but also by NaNO_2. Hexosamine-containing material was also solubilized by these reagents. However, nitrite, unlike lysozyme, released a considerable amount of free hexosamine as well. These results suggest that nitrite-induced germination may involve an interaction of sodium nitrite as nitrous acid with some component of the cortex. A possible mechanism of nitrite-induced germination is discussed.     

Methemoglobin reductase activity and in vitro sensitivity towards oxidant induced methemoglobinemia in Swiss mice and Beagle dogs erythrocytes

The NADH methemoglobin-reductase (EC 1.6.2.2) is mainly responsible for the maintenance of hemoglobin in its reduced and active state. The present study reveals the comparative status of this enzyme in normal Beagle dogs, rats, mice, mastomys and hamsters erythrocytes. The spectrophotometric and electrophoretic determinations showed that the above mentioned enzyme was deficient in the Beagle dog's erythrocytes. Furthermore, in vitro studies on the sensitivity of these rodents and Beagle dogs hemolysate towards oxidants, like primaquine and sodium nitrate, depicted a higher level of methemoglobin formation in the Beagle dogs hemolysate as compared to that of the rodent species. The deficiency of methemoglobin reductase in Beagle dogs erythrocytes could be responsible for their increased sensitivity towards oxidant induced methemoglobinemia.     

Successful Treatment of Epilepsy with Serotonin Reuptake Inhibitors: Proposed Mechanism

The widely used antidepressants Specific Serotonin Reuptake Inhibitors (SSRI) have been tried with success as anticonvulsants in cases of nonsymptomatic epilepsy. This attempt was performed on the basis of experimental data suggesting the involvement of impairments of the serotonin system in the genesis of epilepsy. This overview summarizes the clinical data and presents biochemical and neurochemical evidences suggesting the mechanism of the therapeutic effects of SSRI in nonsymptomatic epilepsy. In particular, studies on blood-borne neutral amino acids and platelet serotonin transporter (SERT) in epileptics suggest: (a) That a decreased brain availability of tryptophan may be related to some types of epilepsy. (b) That reduction of the density of SERT may be a homeostatic reaction in the brain following epileptic seizures.     

Enlargement of Escherichia coli After Bacteriophage Infection II. Proposed Mechanism

Division of Escherichia coli was stopped and mean cellular volume was increased after infection with T-even phage. This host cell enlargement was temperature-dependent, cyanide-sensitive, and stable in the presence of hypertonic medium. Enlargement ceased at about the same time that energy metabolism ceased. Initially, enlargement was accompanied by a decrease in mean cell density. Tritiated 2, 6-diaminopimelic acid was accumulated and incorporated into cold acid-insoluble material at the preinfection rate. These findings suggest that the effect on host cell size is only in part an osmotic phenomenon and that it also reflects continued growth of the surface of the infected cell in the absence of cell division.     

Proton Hopping: A Proposed Mechanism for Myelinated Axon Nerve Impulses

Myelinated axon nerve impulses travel 100 times more rapidly than impulses in non‐myelinated axons. Increased speed is currently believed to be due to ‘hopping’ or ‘saltatory propagation’ along the axon, but the mechanism by which impulses flow has never been adequately explained. We have used modeling approaches to simulate a role for proton hopping in the space between the plasma membrane and myelin sheath as the mechanism of nerve action‐potential flow.     

Fluorescence Decay Time Distribution Analysis Reveals Two Types of Binding Sites for 1,8-Anilinonaphthalene Sulfonate in Native Human Oxyhemoglobin

Binding of 1,8-anilinonaphthalene sulfonate (1,8-ANS) with native human oxyhemoglobin (Hb) in 50 mM potassium phosphate buffer (pH 7.4) was studied by steady state fluorescence spectroscopy and by laser spectrofluorimetry with subnanosecond time resolution. The distribution of fluorescence decay times and parameters of two- and three-exponential deconvolution of the fluorescence kinetics of 1,8-ANS in Hb solution demonstrate that the emission at wavelengths _em of 455-600 nm is not single-exponential and has components with mean decay times <0.5, 3.1-5.5, and 12.4-15.1 nsec with the amplitudes depending on the emission wavelength. Analysis of time-resolved fluorescence spectra shows that the shortest-lived component should be assigned to 1,8-ANS molecules in the aqueous medium, whereas the two longer-lived components are assigned to two types of binding sites for 1,8-ANS in the Hb molecule characterized by different polarity and accessibility to water molecules.     

Magnetic Entropy as a Proposed Gating Mechanism for Magnetogenetic Ion Channels

Magnetically sensitive ion channels would allow researchers to better study how specific brain cells affect behavior in freely moving animals; however, recent reports of “magnetogenetic” ion channels based on biogenic ferritin nanoparticles have been questioned because known biophysical mechanisms cannot explain experimental observations. Here, we reproduce a weak magnetically mediated calcium response in HEK cells expressing a previously published TRPV4-ferritin fusion protein. We find that this magnetic sensitivity is attenuated when we reduce the temperature sensitivity of the channel but not when we reduce the mechanical sensitivity of the channel, suggesting that the magnetic sensitivity of this channel is thermally mediated. As a potential mechanism for this thermally mediated magnetic response, we propose that changes in the magnetic entropy of the ferritin particle can generate heat via the magnetocaloric effect and consequently gate the associated temperature-sensitive ion channel. Unlike other forms of magnetic heating, the magnetocaloric mechanism can cool magnetic particles during demagnetization. To test this prediction, we constructed a magnetogenetic channel based on the cold-sensitive TRPM8 channel. Our observation of a magnetic response in cold-gated channels is consistent with the magnetocaloric hypothesis. Together, these new data and our proposed mechanism of action provide additional resources for understanding how ion channels could be activated by low-frequency magnetic fields.     

铜型亚硝酸还原酶的电子传递模式及催化机理研究进展

亚硝酸还原酶(NiRs)是反硝化过程中的关键作用酶,它使NO2-转变成NO,减轻了水体中的氮污染.根据辅基的不同,NiRs分为血红素cd1型亚硝酸还原酶(cd1-NiRs)和铜型亚硝酸还原酶(Cu-NiRs)两种.Cu-NiRs呈三聚体结构,每个单体都含有两种类型的铜原子,它们在酶催化过程中起着传递电子的重要作用.在催化过程中,Cu-NiRs中部分残基的结构变化有利于催化反应的进行.此文结合Cu-NiRs的最新研究成果,从整体上对其结构特点、电子传递过程及催化机理等研究作了系统地阐述.     

Catalytic Mechanism of Bleomycin N-Acetyltransferase Proposed on the Basis of Its Crystal Structure

Bleomycin (Bm) N-acetyltransferase, BAT, is a self-resistance determinant in Bm-producing Streptomyces verticillus ATCC15003. In our present study, we crystallized BAT under both a terrestrial and a microgravity environment in the International Space Station. In addition to substrate-free BAT, the crystal structures of BAT in a binary complex with CoA and in a ternary complex with Bm and CoA were determined. BAT forms a dimer structure via interaction of its C-terminal domains in the monomers. However, each N-terminal domain in the dimer is positioned without mutual interaction. The tunnel observed in the N-terminal domain of BAT has two entrances: one that adopts a wide funnel-like structure necessary to accommodate the metal-binding domain of Bm, and another narrow entrance that accommodates acetyl-CoA (AcCoA). A groove formed on the dimer interface of two BAT C-terminal domains accommodates the DNA-binding domain of Bm. In a ternary complex of BAT, BmA₂, and CoA, a thiol group of CoA is positioned near the primary amine of Bm at the midpoint of the tunnel. This proximity ensures efficient transfer of an acetyl group from AcCoA to the primary amine of Bm. Based on the BAT crystal structure and the enzymatic kinetic study, we propose that the catalytic mode of BAT takes an ordered-like mechanism.     

Proposed Mechanism of Inheritance and Expression of the Human Fragile-X Syndrome of Mental Retardation

A mechanism is proposed for the inheritance and expression of the fragile-X-linked syndrome of mental retardation in humans. Two independent events are required for expression of the syndrome: the fragile-X mutation, and X chromosome inactivation in pre-oogonial cells. The fragile-X mutation at site Xq27 has little or no effect until the chromosome is inactivated in a female as part of the process of dosage compensation. At a stage where the inactivated X chromosome would normally be reactivated in preparation for oogenesis, the mutation results in a local block to the reactivation process. This block to reactivation leads to mental retardation in progeny by reducing the level of products from the unreactivated Xq27 region in male cells, and, for a heterozygous female, in somatic cells in which the normal X chromosome has been inactivated. Published data relevant to this proposed mechanism are discussed.     

Proposed Mechanism of Antibacterial Activity of Glutathione by Inhibition of the d-Alanyl-d-alanine Carboxypeptidase Enzyme

International Journal of Peptide Research and Therapeutics - Several reports have suggested that glutathione (GSH) has antibacterial activity. However, the mechanism by which GSH inhibits microbial...     

Methemoglobinemia and acidosis during the first week of life

Recently attention has been called on the possible role of acidosis in the increased methemoglobin formation in the erythrocyte of newborn infant. In the present paper the relations between acidosis and methemoglobin content in the red cells of newborns has been investigated. No significant differences between the percent of methemoglobin in the normal newborns and percent of methemoglobin in the newborns with acidosis has been found. In addition, no correlations between the base excess and percent of methemoglobin has been observed. On the contrary, two newborns with low glucose-6-phosphate dehydrogenase activity demonstrated a significantly increased methemoglobin content in their red cells. The results of our study do not confirm a key role of acidosis in the mechanism of methemoglobin formation in the neonate. It is likely than impairment of red cell metabolism should be the main factor in the formation of methemoglobin in the first days of life.     

A microtiter-plate assay of nitric oxide synthase activity

We describe here a microtiter-plate assay for measuring nitric oxide synthase (NOS) activity by utilizing the spectral shift in optical absorbence between the wavelengths 405 and 420 nm on conversion of oxyhemoglobin to methemoglobin by nitric oxide (NO). This is a high-throughput assay permitting 96 or 384 simultaneous kinetic measurements and is ideal for the study of NOS inhibitors and their time dependence. It is also possible to measure enzyme rates under different conditions simultaneously for the study of the cofactor and substrate dependence of NOS preparations. The assay requires approximately 10 pmol/min of NOS activity to achieve a 1moD/min rate.     

Kinetics of sickle hemoglobin polymerization. I. Studies using temperature-jump and laser photolysis techniques

Using a combination of laser photolysis and temperature-jump techniques, the kinetics of hemoglobin S polymerization have been studied over a wide range of delay times (10(-3) to 10(5)s), concentrations (0.2 to 0.4 g/cm3) and temperatures (5 to 50 degrees C). A slow temperature-jump technique was used to induce polymerization in samples with delay times between 10(2) seconds and 10(5) seconds by heating a solution of completely deoxygenated hemoglobin S. For samples with shorter delay times, polymerization was induced by photodissociating the carbon monoxide complex in small volumes (10(-9) cm3) using a microspectrophotometer equipped with a cw argon ion laser. The photolysis technique is described in some detail because of its importance in studying hemoglobin S polymerization at physiological concentrations and temperatures. In order, to establish conditions for complete photodissociation with minimal laser heating, a series of control experiments on normal human hemoglobin was performed and theoretically modeled. The concentration dependence of the tenth time is found to decrease with increasing hemoglobin S concentration. In the range 0.2 to 0.3 g/cm3, the tenth time varies as the 36th power of the hemoglobin S concentration, while in the range 0.3 to 0.4 g/cm3 it decreases to 16th power. As the tenth times become shorter, the progress curves broaden, with the onset of polymerization becoming less abrupt. For tenth times greater than about 30 seconds, measurements with the laser photolysis technique on small volumes yield highly irreproducible tenth times, but superimposable progress curves, indicating stochastic behavior. The initial part of the progress curves from both temperature-jump and laser photolysis experiments is well fit with an equation for the concentration of polymerized monomer, delta (t) = A[cosh (Bt) -1], which results from integration of the linearized rate equations for the double nucleation mechanism described in the accompanying paper (Ferrone et al., 1985). The dependence of the parameters A and B on temperature and concentration is obtained from fitting over 300 progress curves. The rate B has a large concentration dependence, varying at 25 degrees C from about 10(-4) S-1 at 0.2 g/cm3 to about 100 s-1 at 0.4 g/cm3.     

Physiology of Rana pipiens Reproductive Behavior: A Proposed Mechanism for Inhibition of the Release Call

SYNOPSIS. Non-gravid femaleRana pipiens clasped by sexuallyactive males emit a release call and thereby signal their unreceptivestatus to the clasping male. The receptive condition of gravidfemales is indicated by absence, or inhibition, of the call.This paper provides evidence that arginine-8 vasotocin (AVT)can inhibit the release call by causing water uptake when afemale is in an aquatic environment, and suggests that watermight exert this effect by means of a mechanical distensionof the body wall or internal pressure. This paper also includesevidence that the ovaries are not necessary for AVT's inhibitionof the call. The literature dealing with the physiological basisof female frog reproductive behavior and the physiological basisof the release call are reviewed.