Human gene therapy.

Somatic cell gene therapy for the correction of many human genetic diseases is now technically possible. We review several methods of gene transfer that have been successfully used in animal studies, and discuss the promise and potential limitations of these methods in the treatment of human genetic diseases.     

Human somatic cell gene therapy

The prelude to successful human somatic gene therapy, i.e. the efficient transfer and expression of a variety of human genes into target cells, has already been accomplished in several systems. Safe methods have been devised to do this using non-viral and viral vectors. Potentially therapeutic genes have been transferred into many accessible cell types, including hematopoietic cells, hepatocytes and cancer cells, in several different approaches to ex vivo gene therapy. Successful in vivo gene therapy requires improvements in tissuetargeting and new vector design, which are already being sought. Gene-transfer protocols have been approved for human use in inherited diseases, cancer and acquired disorders. Althouth the results of these trials to date have been somewhat disappointing, human somatic cell gene therapy promises to be an effective addition to the arsenal of approaches to the therapy of many human diseases in the 21st century if not sooner.     

Chemical cues, defence metabolites and the shaping of pelagic interspecific interactions

Several observations and model calculations suggest that chemically mediated interactions can structure planktonic food webs. However, only recently have improvements in chemical methods, coupled with ecological assays, led to the characterization of chemical cues that affect the behaviour and/or physiology of planktonic organisms. We are currently beginning to elucidate if or how chemical signals can directly affect the interactions between species and even shape complex community structures in aquatic systems. Here, we highlight recent research on the nature and action of chemical signals in the pelagic marine and freshwater environments, with an emphasis on kairomones and defence metabolites.     

Chemical defence in chrysomelid eggs and neonate larvae

ABSTRACT. Eggs and neonate larvae of chrysomelid beetles (sub-tribes Chrysomelina and Phyllodectina) were investigated for the presence of defensive substances.
The two isoxazolinone glucosides (compounds 1 and 2), characteristic of the adult defence secretion, were detected in the eggs of all studied species. Compound 2, containing a nitropropionate, is always present in concentrations (above 10^-2 M), which are highly deterrent to the ant Myrmica rubra. This compound is not at all or only slightly toxic to ants at 10^-2 M. Compound 1, devoid of nitropropionate, is a minor constituent, and is neither deterrent nor toxic to ants.
The five Chrysomela species studied and Phratora vitellinae also sequester salicin in their eggs in amounts highly deterrent and toxic to ants. A single Chrysomela egg often contains enough salicin to kill an ant. While the isoxazolinones are discarded with the egg shells, salicin is used by neonate larvae as a precursor for the production of salicylaldehyde in the thoracic defence glands, already functional at hatching. No salicin could be detected in the eggs of those species whose larvae produce cyclopentanoid monoterpenes, even if they feed on Salicaceae. No larva of any species seems to be able to produce detectable amounts of monoterpenes at birth. A very early defence, possible only in those species using salicin as the precursor for their defensive secretion, could be highly advantageous in protecting the clustered larvae during the long process of hatching and in avoiding cannibalism between siblings.
Only trace amounts of oleic acid were found in the eggs of Gastrophysa viridula , in contrast to previous reports on its presence in large quantities in the American G. cyanea.     

Chemical defence,offence and alliance in ants–aphids–ladybirds relationships

Chemicals, which mediate the interactions between aphids, ladybirds and ants, are reviewed. Special emphasis is laid on autogenous and plant-derived chemical defence in aphids and ladybirds. Evidences for chemical cues used during foraging and oviposition in ladybirds are assessed. Possible mutualistic interaction between plants and the third trophic level is illustrated by the as yet few reports of indirect plant-defence volatiles induced by aphids or coccids attracting parasitoids or ladybirds. The use of chemical signals allowing aphid parasitoids or ladybirds to squeeze into ant–aphid mutualistic association is briefly described. Questions are raised and hypotheses suggested which could stimulate further research on aphid host-plant influence on ladybird foraging behaviour and fitness, and on the cues used by aphid-web partners for their mutual recognition.     

Gamma-glutamyl transferases: A structural,mechanistic and physiological perspective

Gamma glutamyl transferases （GGT） are highly conserved enzymes that occur from bacteria to humans. They remove terminal y-glutamyl residue from peptides and amides. GGTs play an important role in the homeostasis of glutathione （a major cellular antioxidant） and in the detoxification of xenobiotics in mammals. They are implicated in diseases like diabetes, inflammation, neurodegenerative diseases and cardiovascular diseases. The physiological role of GGTs in bacteria is still unclear. Nothing is known about the basis for the strong conservation of the enzyme across the living system. The review focuses on the enzyme＇s physiology, chemistry and structural properties of the enzyme with emphasis on the evolutionary relationships. The available data indicate that the members of the GGT family share common structural features but are functionally heterogenous.     

The Microbiota,Chemical Symbiosis,and Human Disease

Our understanding of mammalian–microbial mutualism has expanded by combing microbial sequencing with evolving molecular and cellular methods, as well as unique model systems. Here, the recent literature linking the microbiota to diseases of three of the key mammalian mucosal epithelial compartments—nasal, lung, and gastrointestinal tract—is reviewed with a focus on new knowledge about the taxa, species, proteins, and chemistry that promote health and impact progression toward disease. The information presented is further organized by specific diseases now associated with the microbiota: Staphylococcus aureus infection and rhinosinusitis in the nasal-sinus mucosa, as well as cystic fibrosis, chronic obstructive pulmonary disorder, and asthma in the pulmonary tissues. For the vast and microbially dynamic gastrointestinal compartment, several disorders are considered, including obesity, atherosclerosis, Crohn's disease, ulcerative colitis, drug toxicity, and even autism. Our appreciation of the chemical symbiosis ongoing between human systems and the microbiota continues to grow and suggests new opportunities for modulating this symbiosis using designed interventions.     

Chemical gene synthesis: strategies, softwares, error corrections, and applications

Chemical synthesis of DNA sequences provides a powerful tool for modifying genes and for studying gene structure, expression and function. Modified genes and consequently protein/enzymes can bridge genomics and proteomics research or facilitate commercial applications of gene and protein technologies. In this review, we will summarize various strategies, designing softwares and error correction methods for chemical gene synthesis, particularly for the synthesis and assembly of long DNA molecules based on polymerase cycling assembly. Also, we will briefly discuss some of the major applications of chemical synthesis of DNA sequences in basic research and applied areas.     

Antiretroviral therapy in the Malawi defence force: access, treatment outcomes and impact on mortality

Background

HIV/AIDS affects all sectors of the population and the defence forces are not exempt. A national survey was conducted in all public and private sectors in Malawi that provide antiretroviral therapy (ART) to determine the uptake of ART by army personnel, their outcomes while on treatment, and the impact of ART on mortality in the Malawi Defence Force.

Methodology/Principal Findings

A retrospective cohort analysis was carried out, collecting data on access and retention on treatment from all 103 public and 38 private sector ART clinics in Malawi, using standardised patient master cards and clinic registers. Observations were censored on December 31^st 2006. Independent data on mortality trends in army personnel from all causes between 2002 and 2006 were available from army records. By December 31^st 2006, there were 85,168 patients ever started on ART in both public and private sectors, of whom 547 (0.7%) were army personnel. Of these, 22% started ART in WHO clinical stage 1 or 2 with a CD4-lymphocyte count of ≤250/mm³ and 78% started in stage 3 or 4. Treatment outcomes of army personnel by December 31^st 2006 were:−365 (67%) alive and on ART at their registration facility, 98 (18%) transferred out to another facility, 71 (13%) dead, 9 (2%) lost to follow-up, and 4 (<1%) stopped treatment. The probability of being alive on ART at 6-, 12- and 18-months was 89.8%, 83.4% and 78.8% respectively. All-cause mortality in army personnel declined dramatically over the five year period from 2002–2006.

Conclusion/Significance

There has been a good access of army personnel to ART during the last five years with excellent outcomes, and this should serve as an example for other defence forces and large companies in the region.     

Bacteriophages and biotechnology: vaccines, gene therapy and antibacterials

In recent years it has been recognized that bacteriophages have several potential applications in the modern biotechnology industry: they have been proposed as delivery vehicles for protein and DNA vaccines; as gene therapy delivery vehicles; as alternatives to antibiotics; for the detection of pathogenic bacteria; and as tools for screening libraries of proteins, peptides or antibodies. This diversity, and the ease of their manipulation and production, means that they have potential uses in research, therapeutics and manufacturing in both the biotechnology and medical fields. It is hoped that the wide range of scientists, clinicians and biotechnologists currently researching or putting phages to practical use are able to pool their knowledge and expertise and thereby accelerate progress towards further development in this exciting field of biotechnology.     

Barn swallow mobbing: Self-defence,collateral kin defence,group defence,or parental care?

We presented stimuli (human and stuffed owl) to a marked population of barn swallows (Hirundo rustica) during the 1980–1982 breeding seasons at the Cranberry Lake Biological Station, Saint Lawrence County, New York. In experiment 1, we presented the stimuli at various distances from active nests during various stages of the reproductive cycle. Both the probability and intensity of mobbing varied during the breeding season, increasing with the probable reproductive value of young of the year and the degree of danger posed to them. Mobbing group sizes varied positively with local nest densities. In experiment 2, we analysed mobbing group structure in greater detail. Barn swallow mobbing groups usually contained active mobbers (those that emitted mobbing calls, and approached the stimulus closely, < 2 m, or even hit it) and passive mobbers (which were silent and flew in circles at greater distances, 2–10 m, from the stimulus). Passive mobbers were a random sample of the local population with respect to sex, age, nest location and reproductive status. This pattern is consistent with a hypothesis that passive mobbing, the less risky type, is a form of self-defence that reduces an actor's chance of being eaten, probably by providing information about the identity or probable behaviour of potential predators. Active mobbers were not a random sample of the source population. Mated birds and especially parents with nestlings were over-represented, while non-breeders, juveniles and incubators rarely mobbed actively. The seasonal changes in mobbing and the identity of active and passive mobbers are inconsistent with hypotheses that mobbing is a form of cooperative group defence or altruism conditioned by reciprocity or kin or group selection. These data are however consistent with other hypotheses, which propose that mobbing benefits the mate or the young. Even though active mobbers may be at risk, they benefit directly by increasing the personal component of their inclusive fitness, probably by alerting mates and young and defending them from predators. In this light the behavioural complex of mobbing appears to be a form of parental care (active) as well as self-defence (passive) and mate defence (active).     

Chemical alarm and defence in the oribatid mite Collohmannia gigantea (Acari: Oribatida)

The multicomponent oil gland secretion of Collohmannia gigantea, a middle-derivative mixonomatan oribatid mite, is demonstrated to possess alarm pheromonal and allomonal properties. Four components of the secretion, namely the monoterpenes neryl formate, neral, geranial and the aromatic 2-hydroxy- 6-methyl-benzaldehyde (2,6-HMBD), showed moderate to strong alarm pheromonal activity in adult mites. Naturally elicited response is due to neral (about 50% of the secretion) and probably 2,6-HMBD (only 5% of the secretion, but strong alarm pheromonal activity). This is the second report of an alarm pheromone in Oribatida. Tridecane and pentadecane (=the hydrocarbon fraction of the secretion) did not evoke evident behavioural reactions, and most likely serve as solvents and spreading agents for the pheromonal-active components. Alarm reactions were characterized by a short recognition phase (waving movements with legs I), followed by shrinking back and panic escape from the scent source. In addition, all six components of the oil gland secretion, including the hydrocarbons, exhibited strong allomonal properties against a model oribatid predator, the scydmaenid beetle, Euconnus (Tetramelus) oblongus. Considering the widespread semiochemical properties of oil gland secretions in astigmatid mites (=a highly derivative oribatid group), these results furnish evidence for a phylogenetically early origin of defensive and communicative roles of oil gland secretions in oribatids. These roles include alarm communication, defence and the production of anti-fungal compounds.     

Bench,bedside, boardroom: negotiating translational gene therapy

This article presents ethnographic material from a London-based group of gene therapists who received the opportunity to trial a device that, its makers claimed, would expedite and improve their cell work. The Vanguard cell processor elicits both enthusiasm and ambivalence from group members, which I seek to understand by examining the group’s current manner of working alongside the device and its purported virtues. I show that cell processing currently involves complex practices of recognition, attention, care, and involvement, which answer to both the liveliness of cells and the experimentality of gene therapy. I read these practices as a well-honed configuration of productive engagements and detachments, which the Vanguard would thoroughly rearticulate. I thus argue that translational gene therapy is a site at which private and academic interests meet, and that translation more generally might be seen as a space where the relational format of science is renegotiated.     

Glutathione transferases,regulators of cellular metabolism and physiology

Background

The cytosolic glutathione transferases (GSTs) comprise a super family of proteins that can be categorized into multiple classes with a mixture of highly specific and overlapping functions.

Scope of review

The review covers the genetics, structure and function of the human cytosolic GSTs with particular attention to their emerging roles in cellular metabolism.

Major conclusions

All the catalytically active GSTs contribute to the glutathione conjugation or glutathione dependant-biotransformation of xenobiotics and many catalyze glutathione peroxidase or thiol transferase reactions. GSTs also catalyze glutathione dependent isomerization reactions required for the synthesis of several prostaglandins and steroid hormones and the catabolism of tyrosine. An increasing body of work has implicated several GSTs in the regulation of cell signaling pathways mediated by stress-activated kinases like Jun N-terminal kinase. In addition, some members of the cytosolic GST family have been shown to form ion channels in intracellular membranes and to modulate ryanodine receptor Ca^2 + channels in skeletal and cardiac muscle.

General significance

In addition to their well established roles in the conjugation and biotransformation of xenobiotics, GSTs have emerged as significant regulators of pathways determining cell proliferation and survival and as regulators of ryanodine receptors that are essential for muscle function. This article is part of a Special Issue entitled Cellular functions of glutathione.     

Molecular medicine: Molecular diagnostics, preventive medicine, and gene therapy

New trends in molecular medicine that have emerged owing to the success of the national Human Genome program are characterized. The major attention is paid to molecular diagnostics, preventive medicine, and gene therapy. Preventive medicine is a product of synthesis of the current notions on genetics and biochemistry of human diseases; it comprises pharmacogenetics, presymptomatic diagnosis, and testing of genes of predisposition to the most frequent multifactor diseases. In the Gene Therapy section, advantages and drawbacks of the main methods of delivery of nucleic acids into the cells are considered; diseases that are attempted to be rectified using gene therapy are listed. Exemplified with Duchenne myodystrophy, the problems encountered in correction of a genetic defect with the aid of foreign genes are considered. Results are summarized for assessing the efficiency of various methods of introduction of dystrophin cDNA (gene gun, liposomes, microspheres, viral oligopeptides, and lactoferrin) conducted on the Duchenne myodystrophy model, mdx mice.