Endogenous relatives of ADP-ribosylating bacterial toxins in mice and men: potential regulators of immune cell function

ADP-ribosylation of proteins, like phosphorylation, is a post-translational modification that can modulate protein function. Bacterial mono (ADP-ribosyl)transferases have been well studied, since potent and clinically important pathogenic exoenzymes such as diphtheria, cholera and pertussis toxins belong to this group. Some of these enzymes interfere with signal transduction mechanisms of host cells, and have become widely used as research tools in cell biology because of their high potency and selectivity. Recently, relatives of these toxins have been cloned from vertebrates. Seven members of a novel multigene family have been identified to date. Surprisingly, all are predicted to be extracellular proteins. Preferred tissues of expression are skeletal and cardiac muscle, testis and hematopoietic cells. ADP-ribosylation of target proteins on the cell surface of T cells and leukocytes have been found to modulate the transmission of extracellular signals to the cell interior.     

Progress in the development of Lactococcus lactis as a recombinant mucosal vaccine delivery system

Statutory authority for retaking absconders from mental hospitals has existed ever since county asylums (the forerunners of mental hospitals) were first built in the nineteenth century. Today under the Mental Health Act, 1983 that 'right' can be exercised by the police, mental hospital staff, approved social workers, etc. This article looks at jurisprudential aspects of that 'right'. It points out that 'right' actually means 'power' (not 'privilege', 'claim' or 'immunity'). In addition it argues that the Mental Health Act, 1983 does only confer a power (rather than impose a duty) to retake absconders from mental hospitals and that there should not be statutory or other imposition of such a duty.     

Progress in recombinant vaccine development against coccidiosis. A review and prospects into the next millennium

The increasing problems encountered by the poultry industry, despite the extensive use of drugs, have emphasised the need for an immunological solution for the economic damage caused by the Eimeria parasite. Although immunity develops relatively fast following a natural infection, to induce protection by using parasite extracts or single antigens appears more difficult. Nevertheless, the development of a vaccine based on defined antigens seems the best solution in the long run. At the VIth International Coccidiosis Conference in 1993 the first promising results were reported from small-scale experiments using recombinant antigens. This review summarises the advances in this field of research from 1993 onwards. Although since then not many reports have been published about the effects of using recombinant. antigens as a vaccine against coccidiosis, a number of interesting new proteins which could be considered good targets for such a vaccine have been described and are referred to herein. Proteins involved in the process of invasion of the host cell by the extracellular parasite are regarded as key components in the developmental cycle of the parasite. These components possibly bind to receptors on the host cell. Interference with this process could be a target of the protective immune response. Progress has also been made in characterising the immune mechanisms activated by infection with the parasite. From experimental mouse models and from studies in chickens, a better insight has been obtained towards the involvement of CD4- and CD8-type T cells in, respectively, the inductor and the effector branch of the immune response, although not all questions have been answered. Several antigens have been selected using T-cell stimulation and cytokine assays and these are reviewed. In a third section, mostly unpublished results of our own experiments dealing with the use of live vectors to present defined antigens such as Ea1A and EaSC2, a parasite refractile body transhydrogenase and a lactate dehydrogenase, respectively, are summarised. Partial protection could be induced using Salmonella typhimurium as a carrier for these antigens, in that the oocyst output was reduced by up to 50% after challenge and weight gain could be improved by 5-10% over non-vaccinated challenged chickens, when tested in a floor-pen trial. Similar results were obtained when these antigens were presented by viral vectors such as Fowlpox virus or Herpes virus of turkey. These data seem to offer good prospects for the accomplishment of a safe and efficaceous vaccine based on recombinant DNA technology. These expectations are corroborated by recent breakthrough in transfection of related parasites such as Plasmodium and Toxoplasma gondii, and by the increasing amount of genomic information becoming available every day, the impact of which cannot even be estimated yet. These new technologies will allow us to solve the complex problems that we once created ourselves.     

Papillomaviruses and cervical cancer: pathogenesis and vaccine development

A subset of human papillomaviruses (HPVs) has been implicated as the principal etiologic agents of cervical cancer. Cervical cancers consistently retain and express two of the viral genes, E6 and E7. Although infection with HPV seems to be necessary, other factors, such as cellular immune function, play an important role in determining whether cervical infection will regress, persist, or progress to cancer. The close relationship between viral infection and cancer makes HPV an attractive target for prophylactic and therapeutic vaccines. Candidate vaccines have been shown to have efficacy in animal models, and human clinical trials are planned or in progress.     

Environmental toxins and behavioral development: A new role for psychological research.

Presents a new multiple-effects model that emphasizes subtle behavioral alteration as an early sign of toxicity and as evidence that a particular chemical agent may produce long-term impairment in susceptible individuals. The permeability of the placenta to a variety of chemical agents and the special sensitivity of the fetus to some of these agents draws attention to prenatal exposure and the need for prospective longitudinal studies of affective, social, and cognitive development in exposed individuals. The multiple-effects model provides a role for the psychologist in teratological diagnosis and research since the measurement of behavioral variation has developed primarily in psychology. Limitations inherent in both experimental animal research and correlational human studies of toxic effects make it necessary for these methodologies to be used in a complementary fashion. The implications of behavioral teratology for the study of human development and the design of protective social policies are discussed. (45 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The optimization of helper T lymphocyte (HTL) function in vaccine development

Helper T lymphocyte (HTL) responses play an important role in the induction of both humoral and cellular immune responses. Therefore, HTL epitopes are likely to be a crucial component of prophylactic and immunotherapeutic vaccines. For this reason, Pan DR helper T cell epitopes (PADRE), engineered to bind most common HLA-DR molecules with high affinity and act as powerful immunogens, were developed. Short linear peptide constructs comprising PADRE and Plasmodium-derived B cell epitopes induced antibody responses comparable to more complex multiple antigen peptides (MAP) constructs in mice. These antibody responses were composed mostly of the IgG subclass, reactive against intact sporozoites, inhibitory of schizont formation in liver invasion assays, and protective against sporozoite challenge in vivo. The PADRE HTL epitope has also been shown to augment the potency of vaccines designed to stimulate a cellular immune response. Using a HBV transgenic murine model, it was found that CTL tolerance was broken by PADRE-CTL epitope lipopeptide, but not by a similar construct containing a conventional HTL epitope. There are a number of prophylactic vaccines that are of limited efficacy, require multiple boosts, and/or confer protection to only a fraction of the immunized population. Also, in the case of virally infected or cancerous cells, new immunotherapeutic vaccines that induce strong cellular immune responses are desirable. Therefore, optimization of HTL function by use of synthetic epitopes such as PADRE or pathogen-derived, broadly crossreactive epitopes holds promise for a new generation of highly efficacious vaccines.     

Anti-insect scorpion toxins: historical account, activities and prospects

Some toxins from scorpion venoms, much more toxic to insects than to other animal classes, possess high affinity to Na+ channels. These anti-insect scorpion toxins have been divided into: 1) alpha toxins which lack strict selectivity for insects, do not compete with following groups of anti-insect toxins, resemble other alpha scorpion toxins by their structure and their ability, as alpha anemone toxins, to prolong insect axonal action potential durations through a drastic slowing down of the Na+ current inactivation, 2) excitatory insect selective scorpion toxins which induce in blowfly larvae an immediate fast paralysis; in isolated cockroach axons, they depolarize and induce a sustained repetitive activity of short (normal) action potentials through a shift of Na+ activation mechanism towards more negative potentials and some decrease of inactivation at these potential values, 3) depressant insect selective neurotoxins which cause a slow progressive flaccid paralysis of larvae, depolarize insect axons and reduce or even suppress evoked action potentials; resting depolarizations which are antagonized by a post-application of TTX, are due to the opening of sodium channels at very negative potential values and to the suppression of their inactivation mechanism. The decrease of the maximal Na+ conductance following flaccid toxin action may be understood if toxin-modified channels opened at very negative potentials values remain open (or re-open) for much longer times than in control conditions and pass by substate less conductant states. Anti-insect scorpion toxins become of major interest into insect neurophysiology and also into insect pest control, due to their specific target sites and to the recent constructions of insecticidal baculovirus expressions of several of these toxins.     

The psychologies of structure, function, and development.

Reviews E. B. Titchner's 1898 analogy between the anatomical, physiological, and morphogenetic aspects of biological research and the structural, functional, and developmental aspects of psychological research. This analogy is applied to contemporary divisions within psychology. In psychology, structural research tends to be described in the cognitive or mentalist vocabulary, and functional research in the behaviorist vocabulary. However, this correlation is accidental. Structural and functional dimensions of psychological problems and their relation to developmental research are examined. It is concluded that much psychological controversy is caused by concern with different problems, and not by different paradigms for studying the same problems. (45 ref.) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Ownership and governance of university teaching hospitals: let form follow function

Under the best of circumstances, the complex decision-making and resource-allocation processes of a state university (and often of a variety of state agencies important to the university) significantly hinder the ability of the university-owned hospital to make changes critical to its financial and, hence, its programmatic success. At worst, as was the case for the University of Maryland Hospital a decade ago, the hospital can become capital-starved and operationally deficient under the bureaucratic mantle of the state and university and find itself unable to respond to the fast-changing market, placing its viability in jeopardy. To remedy this situation at the University of Maryland Hospital, in 1984 the state created a separate not-for-profit corporation, the University of Maryland Medical System ("the Medical system"), governed by its own board of directors, with a mandate to assure sound business practices, outstanding patient care, access to patients from across the state for tertiary care, access for the local disadvantaged community for comprehensive care, and attention to the academic mission of the university and its school of medicine. The results include strong financial performance, the ability to recapitalize outmoded facilities and technology, growth of strong programs, and the recruitment of excellent chairs and faculty. The Medical System's success suggests that university teaching hospitals, which necessarily depend on patient care revenues, may best be served by (1) removing them from university governance, thus allowing them to give primacy to their mission of patient care, and (2) removing them from state ownership, thus allowing them to use sound business practices in the competitive health care environment. The challenge under this arrangement is to ensure that the teaching hospitals can still support the educational and research programs that distinguish them. By establishing its independent, actively involved board of directors, the Medical System has successfully responded to this challenge.     

Aspects of masticatory form and function in common tree shrews, Tupaia glis

Tree shrews have relatively primitive tribosphenic molars that are apparently similar to those of basal eutherians; thus, these animals have been used as a model to describe mastication in early mammals. In this study the gross morphology of the bony skull, joints, dentition, and muscles of mastication are related to potential jaw movements and cuspal relationships. Potential for complex mandibular movements is indicated by a mobile mandibular symphysis, shallow mandibular fossa that is large compared to its resident condyle, and relatively loose temporomandibular joint ligaments. Abrasive tooth wear is noticeable, and is most marked at the first molars and buccal aspects of the upper cheek teeth distal to P2. Muscle morphology is basically similar to that previously described for Tupaia minor and Ptilocercus lowii. However, in T. glis, an intraorbital part of deep temporalis has the potential for inducing lingual translation of its dentary, and the large medial pterygoid has extended its origin anteriorly to the floor of the orbit, which would enhance protrusion. The importance of the tongue and hyoid muscles during mastication is suggested by broadly expanded anterior bellies of digastrics, which may assist mylohyoids in tensing the floor of the mouth during forceful tongue actions, and by preliminary electromyography, which suggests that masticatory muscles alone cannot fully account for jaw movements in this species.     

Avian gut-associated immune system: implication in coccidial vaccine development

Host responses to coccidia are complex and involve both humoral and cellular immune mechanisms. Convincing evidence exists for a major role of cell-mediated immunity in anticoccidial resistance. With an increasing need for the development of coccidial vaccine, understanding of the intestinal immune system is crucial. To provide increased insight into the immune mechanisms involving mucosal immune response to coccidia, cellular events involved in the maturation of intestinal immune system were studied. Postnatal development of various T lymphocyte subpopulations expressing CD3, CD8, CD4, and antigen-specific T cell receptor (TCR) heterodimers expressing gamma delta (TCR1) or alpha beta (TCR2) was investigated in chickens. Intraepithelial lymphocytes (IEL) expressing the CD8 antigen increased gradually following hatching and subsequently declined with age. The CD4+ cells represented a minor subpopulation among the IEL. The ratios of IEL T cells expressing gamma delta (TCR1) or alpha beta (TCR2) in the intraepithelium of chickens gradually increased after hatching. These results indicate maturational changes of intestinal immune system that should be considered in the development of vaccine against coccidia.     

Bacterial dehalogenases: biochemistry, genetics, and biotechnological applications

This review is a survey of bacterial dehalogenases that catalyze the cleavage of halogen substituents from haloaromatics, haloalkanes, haloalcohols, and haloalkanoic acids. Concerning the enzymatic cleavage of the carbon-halogen bond, seven mechanisms of dehalogenation are known, namely, reductive, oxygenolytic, hydrolytic, and thiolytic dehalogenation; intramolecular nucleophilic displacement; dehydrohalogenation; and hydration. Spontaneous dehalogenation reactions may occur as a result of chemical decomposition of unstable primary products of an unassociated enzyme reaction, and fortuitous dehalogenation can result from the action of broad-specificity enzymes converting halogenated analogs of their natural substrate. Reductive dehalogenation either is catalyzed by a specific dehalogenase or may be mediated by free or enzyme-bound transition metal cofactors (porphyrins, corrins). Desulfomonile tiedjei DCB-1 couples energy conservation to a reductive dechlorination reaction. The biochemistry and genetics of oxygenolytic and hydrolytic haloaromatic dehalogenases are discussed. Concerning the haloalkanes, oxygenases, glutathione S-transferases, halidohydrolases, and dehydrohalogenases are involved in the dehalogenation of different haloalkane compounds. The epoxide-forming halohydrin hydrogen halide lyases form a distinct class of dehalogenases. The dehalogenation of alpha-halosubstituted alkanoic acids is catalyzed by halidohydrolases, which, according to their substrate and inhibitor specificity and mode of product formation, are placed into distinct mechanistic groups. beta-Halosubstituted alkanoic acids are dehalogenated by halidohydrolases acting on the coenzyme A ester of the beta-haloalkanoic acid. Microbial systems offer a versatile potential for biotechnological applications. Because of their enantiomer selectivity, some dehalogenases are used as industrial biocatalysts for the synthesis of chiral compounds. The application of dehalogenases or bacterial strains in environmental protection technologies is discussed in detail.     

Mouse T cell membrane proteins Rt6-1 and Rt6-2 are arginine/protein mono(ADPribosyl)transferases and share secondary structure motifs with ADP-ribosylating bacterial toxins

Mono ADP-ribosylation is a posttranslational protein modification that has been implicated in the regulation of key biological functions in bacteria as well as in animals. Recently, the first cDNAs for eucaryotic mono(ADPribosyl)transferases were cloned and found to exhibit significant sequence similarity only to one other known protein, the T cell differentiation antigen Rt6. In this paper we describe secondary structure analyses of Rt6 and related proteins and show conserved structure motifs and amino acid residues consistent with a common ancestry of these eucaryotic proteins and bacterial ADP-ribosyltransferases. Moreover, we have expressed soluble mouse Rt6-1 and Rt6-2 gene products in which C-terminal tags (FLAG-His6) replace the native glycosylphosphatidylinositol anchor signal sequences. Purified recombinant Rt6-2, but not Rt6-1, shows NAD+ glycohydrolase activity, which is inhibited by the arginine analogue agmatine. Immunoprecipitation of recombinant Rt6-1 and Rt6-2 with anti-FLAG M2 antibody followed by incubation with [32P]NAD+ leads to rapid and covalent incorporation of radioactivity into the light chain of the M2 antibody. The bound label is resistant to treatment with HgCl2 but sensitive to NH2OH, characteristic of arginine-linked ADP-ribosylation. These results demonstrate that Rt6-1 and RT6-2 possess the enzymatic activities typical for NAD+-dependent arginine/protein mono(ADPribosyl)transferases (EC 2.4.2.31). They are the first such enzymes to be molecularly characterized in the immune system.     

Boosting with recombinant vaccinia increases immunogenicity and protective efficacy of malaria DNA vaccine

To enhance the efficacy of DNA malaria vaccines, we evaluated the effect on protection of immunizing with various combinations of DNA, recombinant vaccinia virus, and a synthetic peptide. Immunization of BALB/c mice with a plasmid expressing Plasmodium yoelii (Py) circumsporozoite protein (CSP) induces H-2Kd-restricted CD8+ cytotoxic T lymphocyte (CTL) responses and CD8+ T cell- and interferon (IFN)-gamma-dependent protection of mice against challenge with Py sporozoites. Immunization with a multiple antigenic peptide, including the only reported H-2Kd-restricted CD8+ T cell epitope on the PyCSP (PyCSP CTL multiple antigenic peptide) and immunization with recombinant vaccinia expressing the PyCSP induced CTL but only modest to minimal protection. Mice were immunized with PyCSP DNA, PyCSP CTL multiple antigenic peptide, or recombinant vaccinia expressing PyCSP, were boosted 9 wk later with the same immunogen or one of the others, and were challenged. Only mice immunized with DNA and boosted with vaccinia PyCSP (D-V) (11/16: 69%) or DNA (D-D) (7/16: 44%) had greater protection (P < 0. 0007) than controls. D-V mice had significantly higher individual levels of antibodies and class I-restricted CTL activity than did D-D mice; IFN-gamma production by ELIspot also was higher in D-V than in D-D mice. In a second experiment, three different groups of D-V mice each had higher levels of protection than did D-D mice, and IFN-gamma production was significantly greater in D-V than in D-D mice. The observation that priming with PyCSP DNA and boosting with vaccinia-PyCSP is more immunogenic and protective than immunizing with PyCSP DNA alone supports consideration of a similar sequential immunization approach in humans.     

Host-pathogen interaction in amebiasis and progress in vaccine development

Entamoeba histolytica, the causative organism of invasive intestinal and extraintestinal amebiasis, infects approximately 50 million people each year, causing an estimated 40 to 100 thousand deaths annually. Because amebae only infect humans and some higher non-human primates, an anti-amebic vaccine could theoretically eradicate the organism. Uncontrolled epidemiologic studies indicate that acquired immunity to amebic infection probably occurs and that such a vaccine might be feasible. Application of molecular biologic techniques has led to rapid progress towards understanding how Entamoeba histolytica causes disease, and to the identification of several amebic proteins associated with virulence. These proteins are now being evaluated as potential vaccine components. Parenteral and oral vaccine preparations containing recombinant amebic proteins have been effective in preventing disease in a gerbil model of amebic liver abscess. Although systemic and mucosal cellular and humoral immunity both appear to play a role in protection against Entamoeba histolytica, the relative importance of each in the human immune response remains unknown. No animal model of intestinal amebiasis currently exists, moreover, so it has been impossible to evaluate protection against colonization and colitis. Further investigation of the fundamental mechanisms by which Entamoeba histolytica causes disease and of the human immune response to amebic infection is necessary to assess the true feasibility of an anti-amebic vaccine.     

Optimizing form and function with the direct posterior composite resin: a case report

The human testis determining factor (SRY) has been cloned from the Y chromosome. This gene is a dominant inducer of male differentiation. Mutations in the SRY gene result in an XY individual developing as a sex reversed phenotypic female. Sex reversal in humans can also be caused by mutations located in autosomal or X-linked loci. One such sex-reversing locus (SRAI) is associated with the developmental disorder campomelic dysplasia (CD). Both these syndromes were mapped to human chromosome 17q by the identification of balanced reciprocal translocations in five unrelated patients. The translocation breakpoint of one such XY-female CD patient was mapped and the region surrounding it cloned. The closest distal marker used to map the translocation breakpoint was the SOX9 gene. Because of the close proximity of this gene to the breakpoint, it was subjected to mutation analysis in patients without overt chromosome rearrangements. Analysis of DNA from these patients and their parents identified de novo mutations in the SOX9 gene in patients with both autosomal sex reversal and CD. This showed that mutations in the SOX9 gene are responsible for both syndromes.     

Function follows form: Activation of shape and function features during object identification.

Most theories of semantic memory characterize knowledge of a given object as comprising a set of semantic features. But how does conceptual activation of these features proceed during object identification? We present the results of a pair of experiments that demonstrate that object recognition is a dynamically unfolding process in which function follows form. We used eye movements to explore whether activating one object's concept leads to the activation of others that share perceptual (shape) or abstract (function) features. Participants viewed 4-picture displays and clicked on the picture corresponding to a heard word. In critical trials, the conceptual representation of 1 of the objects in the display was similar in shape or function (i.e., its purpose) to the heard word. Importantly, this similarity was not apparent in the visual depictions (e.g., for the target Frisbee, the shape-related object was a triangular slice of pizza, a shape that a Frisbee cannot take); preferential fixations on the related object were therefore attributable to overlap of the conceptual representations on the relevant features. We observed relatedness effects for both shape and function, but shape effects occurred earlier than function effects. We discuss the implications of these findings for current accounts of the representation of semantic memory. (PsycINFO Database Record (c) 2011 APA, all rights reserved)