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Biomedical subjects

G Hong

Publications and source records attributed to G Hong.

10 recordsLinked to original sources

In vitro replication of adeno-associated virus DNA.

An in vitro assay for adeno-associated virus (AAV) DNA replication has been developed. The substrate is a plasmid containing the duplex form of AAV DNA in pBR322. The AAV insert is excised or rescued from the plasmid by extracts of uninfected cells. Replication was assayed by production of full-length excised AAV DNA resistant to Dpn I digestion. The following results were obtained. (i) Only extracts of cells coinfected with AAV and adenovirus replicated the excised insert. (ii) Density label experiments showed semiconservative replication. (iii) Only the excised AAV insert was replicated; pBR322 sequences were not. (iv) Replication was dependent on the presence of the AAV terminal repeat. (v) If the terminal 55 bases were deleted from both ends of the AAV insert, no rescue took place: replication occurred and both AAV and pBR322 sequences were replicated. We conclude that the AAV terminal repeat is essential for DNA replication but that under some conditions an initiation mechanism that does not involve hairpin priming may be used.

Adenoviruses, Human

Training in basic microsurgical techniques without experiments involving animals.

Until recently, anaesthetised rats have been the usual material employed for learning basic microvascular and microneurosurgical techniques. However, ethical considerations, the costs involved and legislation controlling experiments with animals allow training in microsurgery for extended periods of time at a few medical centres only. This paper reports on our experience of an alternative training programme largely conducted without using live animals. As the basic material we selected legs of slaughtered pigs. According to the developing skill of the trainee, basic microsurgical techniques can be practised on arteries and veins of various sizes in these legs. To verify positive results, the vessels are subsequently perfused with human blood under pressure. The model described is particularly suited to the acquiring of skills in microneurosurgery. Mono-, oligo- and polyfascicular nerves, structurally similar to the configurations found in human extremities, are found in pig legs. The ever-increasing importance of microsurgery in modern medicine requires more and more surgeons and orthopaedists to familiarize themselves with these techniques. The model we propose for teaching and training substantially facilitate such further professional training in an efficient way, and at the same time allows a substantial reduction in the number of experiments conducted on animals.

Animal Testing Alternatives

Intragenomic movement and concerted evolution of satellite DNA in Peromyscus: evidence from in situ hybridization.

Four DNA probes isolated from Peromyscus leucopus were used to examine intra- and interspecific variation in the chromosomal location of satellite DNA in the genus Peromyscus. All four probes hybridized to the centromeric regions of all chromosomes in all species of Peromyscus examined but did not hybridize to the majority of heterochromatic regions in closely related non-Peromyscus species. One probe contains a nonsatellite repetitive sequence. The implications of these data to the evolution of genome organization are discussed.

Animals

The reverse DNA sequencing using Bst DNA polymerase.

The reverse DNA sequencing (RDS) [1] is a rapid method used to check the DNA sequences by sequencing them from the opposite orientation. Because the RDS is basically a double stranded sequencing, the quality of the sequence patterns so obtained generally is not as good as those obtained by the single stranded sequencing, and extra bands and higher background are produced more frequently. This paper shows that the RDS could now generate as good sequence patterns as those obtained by sequencing on the single stranded DNA template if Bst DNA polymerase instead of the conventional enzymes, such as the Klenow enzyme, was used in the RDS. Bst DNA polymerase is heat stable (optimum reaction temperature 65 degrees C) and has recently been successfully used in the conventional DNA sequencing. The RDS has recently been further simplified to meet the need of large DNA sequencing projects such as the human genome project. The combination of the simplified RDS and the use of Bst polymerase should be expected to facilitate greatly the work on sequence confirmation and correction.

Base Sequence

[A realistic practice model for learning microneural surgery without experiments on live animals].

The increasing importance of microneurosurgery in hand and plastic surgery makes it necessary for surgeons to familiarize themselves with these techniques. In most training programs, anesthetized rats have been used to learn basic microneurosurgical techniques. Ethical considerations, the costs involved, and the legislation concerning experiments with live animals permit extensive training in microneurosurgery in only a few medical centers. This paper relates our experience with an alternative training program not involving live animals. Instead, we chose the legs of slaughtered pigs. Mono-, oligo-, and poly-fascicular nerves, structurally similar to the configurations found in human extremities, can be found in pig legs. Neurolysis, primary nerve sutures, and nerve transplantations can be practiced.

Animal Testing Alternatives

Effects of Chinese medicinal herbs on hemorheology in diabetics.

There was a statistically significant increase in whole blood viscosity, plasma viscosity, erythrocyte filterability index and hematocrit in diabetics with "Qi Yin deficiency and blood stasis" syndrome. After treatment with "Yi Qi Yang Yin Hou Xue" prescription there was a statistically significant improvement in hemorheological properties and a slight decrease in fasting blood glucose.

Adult

Logical memory subtest of the Wechsler Memory Scale: age and education norms and alternate-form reliability of two scoring systems.

The Logical Memory (LM) subtest of the Wechsler Memory Scale has been characterized by imprecise scoring instructions which can make data interpretation and study comparisons difficult. A total of 339 adults, from 18 to 83 years old, took either Form I or Form II of the LM. Verbal recall of the story passages was evaluated using gist and verbatim scoring systems. Interrater reliability was very high for both scoring approaches. The two forms were equivalent for gist recall. However, verbatim recall of Form I was more difficult than Form II because the former consists of more words to remember. Recall was related more to educational level than to age. For both gist and verbatim scoring, age and education norms were generated for immediate, delayed, and 24-h recall.

Adolescent

Cardiac glycosides. 6. Gitoxigenin C16 acetates, formates, methoxycarbonates, and digitoxosides. Synthesis and Na+,K+-ATPase inhibitory activities.

A series of 17 gitoxigenin 16 beta-formates, acetates, and methoxycarbonates was synthesized, including their 3 beta-acetates, formates, and digitoxosides. A 16 beta-formate group was generally found to increase activity 30 times, a 16 beta-acetate group 9-12 times, while a 16 beta-methoxycarbonate decreased activity by two-thirds. 3 beta-Formates and acetates had little effect on activity by themselves, but sometimes reduced the activity-increasing properties of 16 beta-formates and acetates. A 3 beta-digitoxoside increases the activity of gitoxigenin by 15 times, but the effect is less if the 16 beta-group is esterified. And finally, a 16-one decreases activity dramatically. These data suggest an important role for C16 esters and possibly the presence of a separate binding site on Na+,K+-ATPase corresponding to the cardenolide C16 position.

Animals

Quality of life following intensive care.

Evaluations of intensive care have largely focused on survival, cost, or functional status; however, these may not be the only outcomes of interest to patients, families, and health care providers. Quality of life is an important consideration in evaluating intensive care because it influences decisions about the use of life-sustaining treatment. A study was conducted to assess the quality of life of survivors of intensive care to determine the relationship of perceived quality of life to other outcomes of intensive care. Patients at least 55 years old who received medical intensive care during 1983 (n = 69) were interviewed to determine social characteristics, functional status, psychological well-being, and their perceived quality of life using a new 11-item Perceived Quality of Life scale. Functional status correlated only moderately with perceived quality of life (r = 0.49, p = 0.0001). Objective measures of patients' material and social resources did not predict satisfaction. To evaluate outcome adequately, it is necessary to assess perceptions of life quality from patients who have received life-sustaining treatment.

Adaptation, Psychological