What investigators need to know about the use of animals.

Investigators conducting research with animal subjects have an ethical and legal responsibility to ensure they are treated humanely. The system of animal research oversight in the United States consists of a framework of federal, state, local, and institutional requirements. Institutions supported by the Public Health Service (PHS) are required to follow the guidelines mandated by the PHS Policy on Humane Care and Use of Laboratory Animals and establish institutional animal care and use committees (IACUC) to oversee animal research activities.

Effects of administration of two growth hormone-releasing hormone plasmids to gilts on sow and litter performance for the subsequent three gestations.

Objective: To determine whether a novel optimized plasmid carrying the porcine growth hormone-releasing hormone (GHRH) wild-type cDNA administered at a lower dose was as effective at eliciting physiologic responses as a commercial GHRH plasmid approved for use in Australia.

Animals: 134 gilts.

Procedures: Estrus was synchronized and gilts were bred.

Gene therapy by electroporation for the treatment of chronic renal failure in companion animals.

Background: Growth hormone-releasing hormone (GHRH) plasmid-based therapy for the treatment of chronic renal failure and its complications was examined. Companion dogs (13.1+/-0.

Techniques in aseptic rodent surgery.

Performing aseptic survival surgery in rodents can be challenging. This unit describes some basic principles to assist clinicians, researchers, and technicians in becoming proficient in performing aseptic rodent surgery.

Effects of plasmid growth hormone-releasing hormone treatment during heat stress.

A gene therapy treatment with plasmid-based growth hormone-releasing hormone (GHRH) delivered by electroporation (EP) was investigated during heat stress; 32 primiparous cows received 2.5 mg of a GHRH-expressing myogenic plasmid (pSP-HV-GHRH), while 20 were designated as controls. Offspring of treated animals showed a reduction in mortality (47%; p < 0.

Growth hormone-releasing hormone plasmid treatment by electroporation decreases offspring mortality over three pregnancies.

LifeTideSW5 is a growth hormone-releasing hormone (GHRH)-expressing plasmid delivered by intramuscular (IM) electroporation (EP), and the first therapeutic plasmid delivered by this physical method to be approved for use in food animals. Gestating sows (n = 997) were treated once with a single 5-mg GHRH-plasmid by EP or served as controls. Data on offspring from three parities subsequent to treatment were collected.

Parameters for DNA vaccination using adaptive constant-current electroporation in mouse and pig models.

Enhancing the expression of DNA vaccines requires that specific conditions of delivery are optimized. We describe experiments using adaptive constant-current electroporation (EP) in mice and pigs examining parameters such as target muscle, delay between plasmid delivery and onset of EP pulses and DNA vaccine formulation; our studies show that concentrated formulations result in better expression and immunogenicity. Furthermore, various conditions of EP that limit the amount of muscle damage were measured.

Double-blinded, Placebo-controlled plasmid GHRH trial for cancer-associated anemia in dogs.

The use of growth hormone releasing hormone (GHRH) plasmid-based therapy to treat companion dogs with spontaneous malignancies and anemia receiving a cancer-specific treatment was examined in a double-blinded, placebo-controlled trial. The dogs (age 10.5 +/- 2.

Delivery of DNA into skeletal muscle in large animals.

Increased transgene expression after plasmid transfer to the skeletal muscle is obtained with electroporation in many species, but optimal conditions for individual species and muscle group are not well defined. Using a muscle-specific plasmid driving the expression of a secreted embryonic alkaline phosphatase (SEAP) reporter gene, we have optimized the electroporation conditions in a large mammal model, i.e.

Plasmid growth hormone releasing hormone therapy in healthy and laminitis-afflicted horses-evaluation and pilot study.

Background: In vivo electroporation dramatically improves the potency of plasmid-mediated therapies, including in large animal models. Laminitis and arthritis are common and debilitating diseases in the horse, as well as humans.

Methods: The effects of growth hormone releasing hormone (GHRH) on healthy horses and on horses with laminitis that were followed for 6 months after a single intramuscular injection and electroporation of 2.

Plasmid-based expression technology using growth hormone releasing hormone: a novel method for physiologically stimulating long-term growth hormone secretion.

Novel DNA-based technologies were recently introduced for various purposes, such as screening of targets identified from genomic projects, shuffled molecules for vaccination, or to direct the in vivo production of hormones and other peptides for therapeutic or preventative applications. We have used a plasmid-based technology to deliver growth hormone releasing hormone (GHRH) to various animal species for screening, toxicology and therapy. A single intramuscular injection of a low dose of plasmid followed by electroporation can ensure that the target species will produce physiological levels of GHRH for extended periods of time, which would replace costly, frequent injections of the recombinant hormone and improve the quality of life and compliance of patients.

Plasmid-based growth hormone-releasing hormone supplementation and its applications.

A single dose of a plasmid expressing growth hormone-releasing hormone (GHRH) has been safely used in a number of animal species and applications to physiologically increase growth hormone and insulin-like growth factor-I for over a year. An array of constructs encoding for analogs of, or species-specific, GHRH has been tested to treat anemia and cachexia associated with cancer and its treatment, and renal failure, as well as to increase immune surveillance and animal welfare. The positive results obtained with plasmid-based GHRH in companion and farm animals may be translated to a number of human applications.

Immune-enhancing effects of growth hormone-releasing hormone delivered by plasmid injection and electroporation.

Growth hormone-releasing hormone (GHRH) is a hypothalamic hormone with both direct and indirect functions in the maintenance of immune status under physiological and pathological conditions. In this study, 52 Holstein heifers were evaluated for the effects of a plasmid-mediated GHRH treatment on their immune function and on the morbidity and mortality of treated animals. In the third trimester of pregnancy, 32 heifers received 2.

Osteoporosis: The dynamic relationship between magnesium and bone mineral density in the heart transplant patient.

Osteoporosis is a common, and potentially severe, complication seen in the heart transplant recipient. Frequently there is loss of bone mineral density prior to transplant that begins the downward spiral to fractures of the femoral neck and vertebrae. Multiple factors are responsible for the development of osteoporosis posttransplant.

Depression and anxiety in the heart transplant patient: a case study.

Depression and anxiety are potential psychological problems that a heart transplant patient may face both before and after transplant. In addition, there is the potential for difficulty in adjusting to life after transplantation. Anxiety and depression and their effects, both pretransplant and posttransplant, are explored.

A case study in Hansen's disease acquired after heart transplant.

Hansen's disease, leprosy, is a chronic infectious disease caused by the acid-fast bacillus Mycobacterium leprae. There are multiple forms of the disease ranging from the relatively benign to the progressive, malignant lepromatous leprosy. There is effective antimicrobial treatment available that is capable of curing the disease.

The management of immunosuppression: the art and the science.

The optimal use of immunosuppressant drugs requires an understanding of their mechanism of action as well as a basic understanding of the biology of transplant rejection and tolerance. The ability to tailor a drug regimen that strikes a fine balance between allograft maintenance and patient well-being demands a sensitivity to the patient's needs and expectations as well. The object of this article is to cover the basic biological principles involved in selecting an immunosuppressant protocol while sharing our experiences with these various regimen.

Optimization of electroporation parameters for the intramuscular delivery of plasmids in pigs.

Increased transgene expression after plasmid transfer to the skeletal muscle is obtained with electroporation in many species, but optimum conditions are not well defined. Using a plasmid with a muscle-specific secreted embryonic alkaline phosphatase (SEAP) gene, we have optimized the electroporation conditions in a large mammal (pig). Parameters tested included electric field intensity, number of pulses, lag time between plasmid injection and electroporation, and plasmid delivery volume.

Maternal GHRH plasmid administration changes pituitary cell lineage and improves progeny growth of pigs.

Previous studies from our laboratory have demonstrated that administration of a myogenic plasmid that encodes a protease-resistant growth hormone-releasing hormone (HV-GHRH) to pregnant rat dams augmented long-term growth in first-generation progeny. In the present study, gilts were injected intra-muscularly at day 85 of gestation with 0, 0.1, 0.

Electrical enhancement of formulated plasmid delivery in animals.

Electroporation has been shown to significantly increase plasmid transfer to the skeletal muscle, but this procedure is also implicated in muscle damage. We are reporting a highly efficient in vivo transfer of a plasmid formulated with poly-(L-glutamate) (PLG) into murine, canine and porcine muscle fibers using electric pulses of low field intensity. In mice and pigs, the use of secreted embryonic alkaline phosphatase (SEAP) as the indicator gene caused increased PLG expression by 2-3 fold compared to naked plasmid; while delivery of a PLG-plasmid formulation to dogs showed a 10-fold increase in serum SEAP levels compared to plasmid alone.