Immune Response to Combination Therapy for Non-Hodgkin Lymphomas

A parametric model of tumor response to combination therapy in the presence of an immune system is described. Synergistic mechanisms which induce tumor regression are simulated with a coupled set of equations. The simulations are first compared to tumor history data obtained with a SCID mouse model to determine key parameters; predictions are then made for an immune-competent animal. The minimum immune cell birth rate relative to malignant B-cell birth rate necessary to induce tumor regression is determined, and optimization of drug combinations in the presence of an immune response is explored. The delayed effect of an immune response relative to drug scheduling is examined, and a mechanism for disease transformation in heterogeneous tumors is proposed.     

Short-term activity cycles in ants: A phase-response curve and phase resetting in worker activity

Single, isolated worker ants are known to become spontaneously active and to respond to interactions with other, active ants. Here I explore the consequences of an interaction between two worker ants on the timing of activity. Isolated worker ants become active after an interval that is characteristic for each individual. The effect of an interaction between two worker ants is strongly dependent on when the interaction takes place. The effect of an interaction is always to decrease the expected interval until the onset of activity. By studying the effect of an interaction on subsequent intervals of activity, it is possible to reject the hypothesis that the change in timing of activity is due to a change in the characteristic period of activity. Rather, the data are consistent with the hypothesis that an interaction causes a phase shift in the normal activity oscillation. The phase-response curve is derived from the observational data. A knowledge of the dynamics of the interactions of individual ants is necessary in order to begin to reconstruct the patterns of colony behavioral activity.     

The structure of the ostracode carapace

The carapace ultrastructure of three Recent [Heterocypris incongruens (Ramdohr), Cypridopsis uidua (Müller), and Conchoecia belgica Müller] and one Cretaceous ostracode species [Cypridea propunctata Sylvester-Bradley] has been studied by means of standard electron microscope techniques. The carapace is shown to be an organic structure divisible into an outer epicuticle, a median exocuticle and an inner endocuticle in the three cypridacean ostracodes examined. In Conchoecia belgica only an outer epicuticle and an inner endocuticle were observed, but this may be due to preservation. The chitin structure of the carapace of calcareous ostracodes is shown to be essentially an interlocking lattice, more coarsely developed in the exocuticle; layered (lamellar) chitin is only present in the selvage and in the connective tissue joining the valves at the hinge. The non-calcareous Conchoecia belgica possesses the layered chitin structure common to other crustaceans and to insects. The relationship of the carapace structure to that present in the decapod and insect exoskeleton is discussed. The technical problems met with during this study are considered in order to outline the difficulties associated with the study of organisms of this size.     

Characterization of melatonin-induced fos-like immunoreactivity in the hypothalamic suprachiasmatic nucleus of the rat.

The hypothalamic suprachiasmatic nucleus (SCN) is primarily responsible for the regulation of circadian rhythmicity. Melatonin, the pineal-derived neurohormone, modulates the rhythmic output of the SCN. Property timed exposure to melatonin is able to induce changes in rhythmic function and thereby entrain circadian rhythms of activity. c-fos is an immediate early gene that is transiently expressed in neurons in response to receptor activation. The ventrolateral portion of the SCN (vSCN) is activated in response to phase-shifting stimuli, an event which is marked by an increase in the expression of c-fos.     

An oxygen-dependent coproporphyrinogen oxidase encoded by the hemF gene of Salmonella typhimurium.

The 8th step in the 10-step heme biosynthetic pathway of Salmonella typhimurium is the oxidation of coproporphyrinogen III to protoporphyrinogen IX. On the basis of genetic studies, we have suggested that this reaction may be catalyzed by either of two different enzymes, an oxygen-dependent one encoded by hemF or an oxygen-independent enzyme encoded by hemN. Here, we report the cloning of the S. typhimurium hemF gene and its DNA sequence. The predicted amino acid sequence of the HemF protein is 44% identical to that of the coproporphyrinogen oxidase encoded by the yeast HEM13 gene. The wild-type S. typhimurium strain LT-2 produces an oxygen-dependent coproporphyrinogen oxidase activity detectable in crude extracts, which is not found in hemF mutants and is overproduced in strains carrying the hemF gene on a multicopy plasmid. the hemF gene is the second gene in an operon with an upstream gene with an unknown function, whose amino acid sequence suggests a relation to amidases involved in cell wall synthesis or remodeling. The upstream gene and hemF are cotranscribed from a promoter which was mapped by primer extension. A weaker, hemF-specific promoter is inferred from the behavior of an omega-Cm insertion mutation in the upstream gene. Although this insertion decreases expression of beta-galactosidase about 7.5-fold when placed upstream of a hemF-lacZ operon fusion, it still allows sufficient HemF expression from an otherwise wild-type construct to confer a Hem+ phenotype. The hemF operon is transcribed clockwise with respect to the genetic map.     

Characteristic features of electron-ion collisions in strong electric fields

The classical motion of an electron in the Coulomb field of an ion and in a uniform external electric field is analyzed. A nondimensionalization method that makes it possible to study electron motion in arbitrarily strong electric fields is proposed. The possible electron trajectories in the plane of motion in a static field are classified. It is noted that, from a practical standpoint, the most interesting trajectories are snakelike trajectories, which are absent in the problem with a weak external field. An adiabatic approximation for transverse electron motions in quasistatic (strong) fields is constructed. A one-dimensional equation of motion is derived that accounts for transverse electron oscillations and the increase in the effective electron mass as an electron approaches an ion. An analytic model is used to calculate the spectra of bremsstrahlung generated by individual electrons. The calculated results are shown to agree well with the results of direct numerical integration of the basic equations. It is predicted that, at frequencies higher than the frequency of the incident light, pronounced peaks can appear in the spectrum of the transverse dipole moment of an electron; as a result, an electron is expected to effectively emit radiation at these frequencies in the direction of the external field.     

Risk assessment for quantitative responses using a mixture model

A problem that frequently occurs in biological experiments with laboratory animals is that some subjects are less susceptible to the treatment than others. A mixture model has traditionally been proposed to describe the distribution of responses in treatment groups for such experiments. Using a mixture dose-response model, we derive an upper confidence limit on additional risk, defined as the excess risk over the background risk due to an added dose. Our focus will be on experiments with continuous responses for which risk is the probability of an adverse effect defined as an event that is extremely rare in controls. The asymptotic distribution of the likelihood ratio statistic is used to obtain the upper confidence limit on additional risk. The method can also be used to derive a benchmark dose corresponding to a specified level of increased risk. The EM algorithm is utilized to find the maximum likelihood estimates of model parameters and an extension of the algorithm is proposed to derive the estimates when the model is subject to a specified level of added risk. An example is used to demonstrate the results, and it is shown that by using the mixture model a more accurate measure of added risk is obtained.     

Identification and estimation algorithm for stochastic neural system. II

The algorithm for identifying the stochastic neural system and estimating the system process which reflects the dynamics of the neural network are presented in this papar. The analogous algorithm has been proposed in our preceding paper (Nakao et al., 1984), which was based on the randomly missed observations of a system process only. Since the previous algorithm mentioned above was subject to an unfavorable effect of consecutively missed observations, to reduce such an effect the algorithm proposed here is designed additionally to observe an intensity process in a neural spike train as the information for the estimation.The algorithm is constructed with the extended Kalman filters because it is naturally expected that a nonlinear and time variant structure is necessary for the filters to realize the observation of an intensity process by means of mapping from a system process to an intensity process. The performance of the algorithm is examined by applying it to some artificial neural systems and also to cat's visual nervous systems. The results in these applications are thought to prove the effectiveness of the algorithm proposed here and its superiority to the algorithm proposed previously.     

Spindle disturbances in mammalian cells. IV. The action of some glutathione-specific agents in V79 Chinese hamster cells, changes in levels of free sulfhydryls and ATP, c-mitosis and effects on DNA metabolism

The glutathione-specific agents diamide, diethyl maleate and 1-chloro-2,4-dinitrobenzene were found to induce a low frequency of c-mitosis (15%) at non-toxic concentrations concomitant with a 30-40% decrease of non-protein sulfhydryls. The frequency of c-mitosis did not increase further with increased concentrations until non-protein sulfhydryl levels were obtained suggesting depletion of reduced glutathione. The observed shape of the concentration-response curve for c-mitosis is particular to these 3 agents and caffeine among 22 different compounds being tested under comparable conditions. This suggests a similar mechanism of action and from what is known about caffeine this mechanism probably involves an impaired control of cytoplasmic free Ca2+. It is speculated that this impairment with the glutathione-specific agents is primarily due to depletion of a particular pool of reduced glutathione. Tertiary butylhydroperoxide which is a substrate for glutathione peroxidase(s) also causes c-mitosis when there is no significant decrease of non-protein sulfhydryls. The c-mitotic response was found to be biphasic with maintained control levels at an intermediate concentration. The humps in the concentration-response curve for c-mitosis appeared coincident with a mitogenic response (G1----S). Since the latter type of effect most probably is Ca2+ dependent and since the spindle is sensitive to Ca2+ it is tentatively suggested that the c-mitotic effect of tertiary butylhydroperoxide is due to an increase of cytoplasmic Ca2+. Measurements performed imply that an increase of glutathione disulfide (diamide) is more inhibitory to uptake and incorporation of thymidine than a decrease of reduced glutathione per se (diethyl maleate). This difference is probably due to secondary effects on pertinent protein sulfhydryls with diamide, one possible target being the ribonucleotide reductase. All compounds were found to cause an increase of ATP with some of the applied concentrations. The results with diethyl maleate suggest that an increase of ATP is favored by an attack on mitochondrial reduced glutathione. The possible analogy between this effect and an increase of ATP and Ap4A in bacteria during oxidative stress is considered.     

Simulation of human sleep: ultradian dynamics of electroencephalographic slow-wave activity

The typical declining trend of electroencephalographic (EEG) slow-wave activity (SWA) within a sleep period is represented in the two-process model of sleep regulation by an exponentially decaying process (Process S). The model has been further elaborated to simulate not only the global changes of SWA, but also the dynamics within non-rapid-eye-movement (non-REM) sleep episodes. In this new model, the initial intraepisodic buildup of SWA is determined by the combined action of an exponentially increasing process and a saturation process, whereas its fall at the end of an episode is due to an exponentially decreasing process. The global declining trend of SWA over consecutive episodes results from the monotonic decay of the intraepisodic saturation level. In contrast to Process S in the two-process model, this decay is not represented by an exponential function, but is proportional to the momentary level of SWA. REM sleep episodes are triggered by an external function. The model allows one to simulate the ultradian pattern of SWA for baseline nights as well as changes induced by a prolonged waking period, a daytime nap, a partial slow-wave sleep deprivation, or an antidepressant drug.     

Evolutionarily stable strategies for a finite population and a variable contest size

This paper presents a generalization of Maynard Smith's concept of an evolutionarily stable strategy (ESS) to cover the cases of a finite population and a variable contest size. Both equilibrium and stability conditions are analysed. The standard Maynard Smith ESS with an infinite population and a contest size of two (pairwise contests) is shown to be a special case of this generalized ESS. An important implication of the generalized ESS is that in finite populations the behaviour of an ESS player is "spiteful", in the sense that an ESS player acts not only to increase his payoff but also to decrease the payoffs of his competitors. The degree of this "spiteful" behaviour is shown to increase with a decrease in the population size, and so is most likely to be observed in small populations. The paper concludes with an extended example: a symmetric two-pure-strategies two-player game for a finite population. It is shown that a mixed strategy ESS is globally stable against invasion by any one type of mutant strategist. The condition for the start of simultaneous invasion by two types of mutant is also given.     

Spontaneous and UV-induced mutations in Escherichia coli K-12 strains with altered or absent DNA polymerase I.

The induction of mutations to valine resistance and to rifampin resistance occurs after UV irradiation in bacteria carrying a deletion through the polA gene (delta polA), showing that DNA polymerase I (PolI) is not an essential enzyme for this process. The PolI deletion strain showed a 7- to 10-fold-higher spontaneous mutation frequency than the wild type. The presence in the deletion strain of the 5'----3' exonuclease fragment on an F' episome caused an additional 10-fold increase in spontaneous mutation frequency, resulting in mutation frequencies on the order of 50- to 100-fold greater than wild type. The mutator effect associated with the 5'----3' exonuclease gene fragment together with much of the effect attributable to the polA deletion was blocked in bacteria carrying a umuC mutation. The mutator activity therefore appears to reflect constitutive SOS induction. Excision-proficient polA deletion strains exhibited increased sensitivity to the lethal effect of UV light which was only partially ameliorated by the presence of polA+ on an F' episome. The UV-induced mutation rate to rifampin resistance was marginally lower in delta polA bacteria than in bacteria carrying the polA+ allele. This effect is unlikely to be caused by the existence of a PolI-dependent mutagenic pathway and is probably an indirect effect caused by an alteration in the pattern of excision repair, since it did not occur in excision-deficient (uvrA) bacteria. An excision-deficient polA deletion strain possessed UV sensitivity similar to that of an isogenic strain carrying polA+ on an F' episome, showing that none of the functions of PolI are needed for postreplication repair in the absence of excision repair. Our data provide no evidence for a pathway of UV mutagenesis dependent on PolI, although it remains an open question whether PolI is able to participate when it is present.     

NikD, an unusual amino acid oxidase essential for nikkomycin biosynthesis: structures of closed and open forms at 1.15 and 1.90 A resolution

NikD is an unusual amino-acid-oxidizing enzyme that contains covalently bound FAD, catalyzes a 4-electron oxidation of piperideine-2-carboxylic acid to picolinate, and plays a critical role in the biosynthesis of nikkomycin antibiotics. Crystal structures of closed and open forms of nikD, a two-domain enzyme, have been determined to resolutions of 1.15 and 1.9 A, respectively. The two forms differ by an 11 degrees rotation of the catalytic domain with respect to the FAD-binding domain. The active site is inaccessible to solvent in the closed form; an endogenous ligand, believed to be picolinate, is bound close to and parallel with the flavin ring, an orientation compatible with redox catalysis. The active site is solvent accessible in the open form, but the picolinate ligand is approximately perpendicular to the flavin ring and a tryptophan is stacked above the flavin ring. NikD also contains a mobile cation binding loop.     

Physical and biological properties of an epsilon-heavy chain mRNA and a kappa-light chain mRNA coding for rat immunoglobulin E

The mRNA coding for epsilon-heavy chain and kappa-light chain have been highly enriched from a rat IgE-producing myeloma, IR-162. Based on denaturing gel analyses, the 20 S epsilon-heavy chain mRNA has an estimated molecular weight of 850,000, equivalent to about 2500 nucleotides. The 14S rat kappa-light chain mRNA has an estimated molecular weight of 410,000, equivalent to about 1200 nucleotides. Only about two-thirds of the length of these mature cytoplasmic rat mRNA code for protein. The 20 S mRNA stimulates the in vitro synthesis of a single major serologically related protein which is large enough to be epsilon-heavy chain. It is unglycosylated and has an apparent molecular weight of about 62,000. The in vivo unglycosylated epsilon-heavy chain, obtained in the presence of tunicamycin, has an apparent molecular weight of about 59,000, compared with about 76,000 for the glycosylated heavy chain of the secreted rat IgE. Therefore, the in vitro synthesized epsilon-heavy chain protein is about Mr = 3000 larger than the in vivo unglycosylated epsilon-heavy chain, equivalent to about 25 extra amino acids. This is consistent with the synthesis of an epsilon-heavy chain putative precursor. Likewise, the 14 S mRNA stimulates the in vitro synthesis of a single putative precursor protein, which is serologically related to kappa chain, is unglycosylated, and is about an extra 20 amino acids. This is the first report on the physical and biological properties of an epsilon-heavy chain mRNA, as well as any rat immunoglobulin mRNA.     

Interaction of melanin with proteins--the importance of an acidic intramelanosomal pH.

Melanin is a highly irregular heteropolymer consisting of monomeric units derived from the enzymatic oxidation of the amino acid tyrosine. The process of melanin formation takes place in specialized acidic organelles (melanosomes) in melanocytes. The process of melanin polymerization requires an alkaline pH in vitro, and therefore, the purpose of an acidic environment in vivo remains a mystery. It is known that melanin is always bound to protein in vivo. It is also seen that polymerization in vitro at an acidic pH necessarily requires the presence of proteins. The effect of various model proteins on melanin synthesis and their interaction with melanin was studied. It was seen that many proteins could increase melanin synthesis at an acidic pH, and that different proteins resulted in the formation of different states of melanin, i.e., a precipitate or a soluble, protein-bound form. We also present evidence to show that soluble protein-bound melanin is present in vivo (in B16 cells as well as in B16 melanoma tissue). An acidic pH appeared to be necessary to ensure the formation of a uniform, very high molecular weight melano-protein complex. The interaction between melanin and proteins appears to be largely charge-dependent as evidenced by zeta potential measurements, and this interaction is also increased in an acidic pH. Thus, it appears that an acidic intramelanosomal pH is essential to ensure maximum interaction between protein and melanin, and also to ensure that all the melanin formed is protein-bound.     

The process of dissolving apolipoprotein A-I in an aqueous buffer

Human plasma apolipoprotein A-I (apoA-I) has been studied in an aqueous solution by the techniques of high performance liquid chromatography (HPLC), circular dichroic spectroscopy, and sedimentation equilibrium ultracentrifugation. The results indicate that an oligomer is formed as an intermediate step of dissolving lysophilized apoA-I. The process of further dissolution of this oligomer is an irreversible, temperature-dependent dissociation. The half-life of this intermediate oligomer is 3 min at 37 degrees C and 80 h at 30 degrees C. The completely dissolved apoA-I in an aqueous buffer self-associates with conformational alteration. The self-association equilibrium is too rapid to be demonstrated by HPLC.     

Competition for chiasmata in diploid and teisomic maize

Summary The addition of an extra chromosome, in particular the short arm of chromosome V, inZea Mays is shown to lead to (a) less competition for chiasmata between the chromosomes, (b) an increase in the number of chiasmata formed. These effects are in accordance with the hypothesis of control by an effective upper limit to the number of chiasmata which may be formed in any nucleus. The addition of an extra chromosome raises the upper limit of the number of possible chiasmata. This is further supported by the fact that the increase in chiasma formation is not confined to the trisomic chromosome but is, as far as can be judged, shared by all the bivalents. The bearing of these findings on chiasma frequency studies of polyploid series is briefly discussed.     

Luminol chemiluminescence: Chemistry,excitation, emitter

The mechanism of luminol chemiluminescence is a special case of nucleophilic addition to carbonyl compounds. The breakdown of the key intermediate, an alpha hydroxy hydroperoxide, produces a peracid ortho to an acyl diazene group. After intramolecular addition of the peracid, the energy from nitrogen expulsion is utilized in the formation of an anti-aromatic endoperoxide. Rupture along the O,O bond leaves a substantial part of the ensuing phthalate in its excited state. The emitter is shown to be a mono-protonated phthalate unaccessible by photoexcitation. The dark reaction is a concerted decomposion of the alpha hydroxy hydroperodixe to yield ground-state phthalate.