Prospects for insulin delivery by ex-vivo somatic cell gene therapy

The principle of insulin delivery by ex-vivo somatic cell gene therapy involves the removal of non-B-cell somatic cells (e.g. fibroblasts) from a diabetic patient, and genetically altering them in vitro to produce and secrete insulin. The cells can be grown in culture and returned to the donor as a source of insulin replacement. Cells modified in this way could be evaluated before implantation, and reserve stocks could be cryopreserved. By using the patient's own cells, the procedure should obviate the need for immunosuppression and overcome the problem of tissue supply, while avoiding a recurrence of cell destruction. Ex-vivo somatic cell gene therapy requires an accessible and robust cell type that is amenable to multiple transfections and subject to controlled proliferation. Special problems associated with the use of non-B-cell somatic cells include the processing of proinsulin to insulin, and the conferment of sensitivity to glucose-stimulated proinsulin biosynthesis and regulated insulin release. Preliminary studies using fibroblasts, pituitary cells, kidney (COS) cells and ovarian (CHO) cells suggest that these challenges could be met, and that ex-vivo somatic cell gene therapy offers a feasible approach to insulin replacement therapy.     

Smoking and Parkinson''s disease: a dose-response relationship

BACKGROUND: Previous studies questioned the link between early childhood anemia and detrimental child development. OBJECTIVE: A population-based study was conducted to examine the association between early childhood anemia and mild or moderate metal retardation at 10 y of age. DESIGN: The present study linked early childhood nutrition data collected by the Special Supplemental Program for Women, Infants, and Children (WIC) and school records. Hemoglobin values were used to determine the relation between anemia in early life and children's placement in special education classes for mild or moderate mental retardation. Subjects were all participants in the WIC program. A computer program was used to link data from birth, WIC, and school records. RESULTS: Logistic regression showed an increased likelihood of mild or moderate mental retardation associated with anemia, independent of birth weight, maternal education, sex, race-ethnicity, the mother's age, or the child's age at entry into the WIC program. CONCLUSION: These findings support the proposition that efforts to prevent mild and moderate mental retardation should include providing children with adequate nutrition during early childhood.     

TOXICITY AND ANTIBACTERIAL EFFECT OF MACE OF MYRISTICA FRAGRANS USED IN MOROCCAN GASTRONOMY: BIOCHEMICAL AND HISTOLOGICAL IMPACT

The aril of the plant Myristica fragrans is one of the most broadly used spices in Moroccan gastronomy. This research demonstrated its antimicrobial activity against both gram-positive and gram-negative bacterial species. Likewise, its toxicity was investigated on Swiss albino mice. Mice were treated orally with 0.003 and 0.3 mg/day during 7 days. Plasmatic markers and antioxidant defense systems were assessed and histological alterations evaluated. Our results showed a significant increase in creatine phosphokinase level. The microscopic evaluation showed that mace induces morphological perturbation in mice's liver. The results also showed an inhibitory effect of glyceraldehyde 3-phosphate dehydrogenase and an important increase in the level of thiobarbituric acid reactive substances, succinate dehydrogenase activities and no change in catalase activities. All of these results show that M. fragrans at 0.3 mg/g in mice affects energy metabolism and oxidative stress.

PRACTICAL APPLICATIONS

Mace can be used as a natural antioxidant and preservative in the food industry to increase the shelf life of foodstuffs.     

Interkey timing in piano performance and typing

In typing, when the fingers executing two successive movements are on the same hand, the time between keystrokes is longer than when the fingers are on different hands. Biomechanical limitations of the hands are thought to account for this difference. The generality of this finding was explored by investigating skilled pianists' performance of two successive notes. Experiment 1 failed to find comparable differences in timing as a function of the hands involved. Experiment 2, employing both a piano production and a typing task, replicated the previous piano performance results, and revealed that the timing differences in typing were limited to letter sequences requiring fore-aft and lateral finger movements. Experiment 3 extended this finding to piano performance. Together, these findings clarify the nature of biomechanical constraints on skilled manual performance.     

Sulfotransferase gene expression in primary cultures of rat hepatocytes

Hepatocyte cultures have been used in pharmacotoxicological studies, and sulfotransferases (ST) are important drug-metabolizing enzymes in liver. The expression of sulfotransferases in hepatocyte cultures has not been examined systematically. In the present study, the mRNA levels of different sulfotransferases in male and female rat hepatocytes were examined by northern-blot analyses. Various culture conditions such as different matrices (collagen, matrigel, collagen sandwich, or co-culture with epithelial cells), medium (Way-mouth's MB 752/1 and Modified Chee's Medium) and glucocorticoid supplementation (dexamethasone, 0.1 microM) were compared. Phenol ST (ST1A1) mRNA levels decreased to about 50% of initial mRNA levels within 10 hr of culture. At 96 hr, ST1A1 mRNA levels were approximately 20% of initial values when cultured on collagen, matrigel or co-culture. The two media did not differ in ability to maintain ST1A1 mRNA levels in the absence of dexamethasone (DEX); however, DEX addition to either medium resulted in ST1A1 mRNA levels greater than 100% of the initial mRNA levels at 96 hr, with the greatest increase observed using the matrigel substratum and Chee's medium. In the absence of DEX, the mRNA levels of N-hydroxy-2-acetylaminoflurene sulfortransferase (ST1C1), estrogen sulfotransferase (ST1E2) and hydroxysteroid sulfotransferase (ST-20/21, ST-40/41, ST-60) fell to approximately 20% of their initial levels within 24 hr, and to less than 5% at 96 hr. The loss of expression of these sulfotransferases was observed with all culture conditions. Addition of DEX to the media resulted in ST-40/41 and ST-60 mRNA expression at 20 and 35% of their initial values, respectively, in cultures maintained on matrigel and Chee's medium at 96 hr. These data suggest that sulfotransferases lose their constitutive expression in hepatocyte culture, but retain their inducibility.     

Indications for the surgical repair of unilateral aural atresia in children

OBJECTIVE: This study aimed to clarify the indications for elective surgical repair of unilateral aural atresia in children and to review the rates of successful repair in the literature. DATA SOURCES: A search of the published English language literature, 1966-1997, was conducted using the key words aural atresia. STUDY SELECTION: Articles were selected on the basis of their inclusion of the authors' indications for surgical repair of aural atresia or the inclusion of surgical series that showed outcomes. DATA EXTRACTION: Articles reviewed in the review had to either provide specific guidelines for surgical repair of unilateral aural atresia or provide postoperative pure-tone averages, air-bone gaps, or speech reception thresholds. DATA SYNTHESIS: The authors compiled the relevant data into summary tables and extracted conclusions from these data. CONCLUSIONS: Elective surgical repair of unilateral aural atresia should only be attempted in children who meet specific anatomic criteria that predict that they are the most likely to benefit from the results of surgery. Otherwise, repair should be delayed until the age at which the patient can make an informed decision, knowing the risks, benefits, and consequences of this difficult surgery. Parents and surgeons must have a realistic expectation of the surgical results and the practiced benefit to be expected with a normal, contralateral ear.     

Saccharide-assisted delivery of cytotoxic liposomes to human malignant cells

The overexpression of lectins by malignant cells was applied for in vitro targeting of liposomes equipped with a saccharide vector and loaded in the lipid phase with a lipid derivative of anticancer agent sarcolysine. The lectin specificity of human leukemia HL-60 and human lung adenocarcinoma ACL cells was revealed by tests with fluorescein-labeled sugar probes. With the help of fluorescent lipid dye it was shown that active saccharide ligands increased the level of the vectored liposome binding to malignant cells by 50-80% as compared to liposomes without vector or with inactive one. The degree of liposome/cell membrane fusion was monitored fluorometrically and was shown to be complete and independent of the vectors. The targeted drug-loaded liposomes had the cytotoxic activity 2-4 times higher as compared to the vector-free ones.