Endogenous tissue engineering: PTH therapy for skeletal repair

Based on its proven anabolic effects on bone in osteoporosis patients, recombinant parathyroid hormone (PTH_1-34) has been evaluated as a potential therapy for skeletal repair. In animals, the effect of PTH_1-34 has been investigated in various skeletal repair models such as fractures, allografting, spinal arthrodesis and distraction osteogenesis. These studies have demonstrated that intermittent PTH_1-34 treatment enhances and accelerates the skeletal repair process via a number of mechanisms, which include effects on mesenchymal stem cells, angiogenesis, chondrogenesis, bone formation and resorption. Furthermore, PTH_1-34 has been shown to enhance bone repair in challenged animal models of aging, inflammatory arthritis and glucocorticoid-induced bone loss. This pre-clinical success has led to off-label clinical use and a number of case reports documenting PTH_1-34 treatment of delayed-unions and non-unions have been published. Although a recently completed phase 2 clinical trial of PTH_1-34 treatment of patients with radius fracture has failed to achieve its primary outcome, largely because of effective healing in the placebo group, several secondary outcomes are statistically significant, highlighting important issues concerning the appropriate patient population for PTH_1-34 therapy in skeletal repair. Here, we review our current knowledge of the effects of PTH_1-34 therapy for bone healing, enumerate several critical unresolved issues (e.g., appropriate dosing regimen and indications) and discuss the long-term potential of this drug as an adjuvant for endogenous tissue engineering.     

Physiological characterization of opine-utilizing rhizobacteria for traits related to plant growth-promoting activity

Thirty-two strains of opine-utilizing rhizobacteria were evaluated for physiological traits which have been related to plant growth-promoting activity. Tests included antibiosis against two bacterial and eight fungal pathogens of potato (Solanum tuberosum L.), production of hydrogen cyanide and fluorescent pigment production. On average, 71 and 12% of the bacteria inhibited the growth of Erwinia carotovora subsp. carotovora and Agrobacterium tumefaciens, respectively. The growth of Botrytis sp. was inhibited by 62% of the bacteria, and half of these produced an inhibition zone of more than 7 mm in diameter. Fusarium solani, Colletotrichum coccodes, Phoma exigua, Verticillium dahliae, F. oxysporum, V. albo-atrum and F. sambucinum were antagonized by 43, 34, 31, 25, 19, 18, and 12% of the bacteria, respectively. Only four strains produce hydrogen cyanide. The inhibition of a plant pathogen was not correlated to the production of fluorescent pigment. No strain produced a hypersensitive reaction whereas only three strains induced soft-rot and two produced polygalacturonase. Some opine-utilizing rhizobacteria were strong inhibitors of all plant pathogens, while most were active against specific plant pathogens.     

Untargeted mutagenesis induced by UV in the lacI gene of Escherichia coli

Summary Using a nonselective method, we have estimated the proportion of untargeted mutations in the lacI gene of E. coli by transferring either irradiated or unirradiated F pro lac plasmids from an excision deficient donor to an excision deficient pro lac deleted recipient that had been irradiated and allowed to induce recA dependent functions for 30 min. We find that about 10 percent of the mutations induced by either 3.5 Jm^-2 or 7 Jm^-2 UV are untargeted.     

New mutations affecting induced mutagenesis in yeast

Previously isolated mutations in baker's yeast, Saccharomyces cerevisiae, that impair induced mutagenesis were all identified with the aid of tests that either exclusively or predominantly detect base-pair substitutions. To avoid this bias, we have screened 11 366 potentially mutant clones for UV-induced reversion of the frameshift allele, his4–38, and have identified 10 mutants that give much reduced yields of revertants. Complementation and recombination tests show that 6 of these carry mutations at the previously known REV1, REV1 and REV3 loci, while the remaining 4 define 3 new genes, REV4 (2 mutations), REV5 and REV6. The rev4 mutations are readily suppressed in many genetic backgrounds and, like the rev5 mutation, impart only a limited deficiency for induced mutagenesis: it is likely, therefore that the REV4⁺ and REV5⁺ gene functions are only remotely concerned with this process. The rev6 mutants have a more general deficiency, however, as well as marked sensitivity to UV and an increased spontaneous mutation rate, properties that suggest the REV6 gene is directly involved in mutation induction. The REV5 gene is located about 1 cM proximal to CYC1 on chromosome X.     

Fine Structure Mapping in Yeast with Sunlamp Radiation

The X-ray mapping procedure of Manney and Mortimer (1964) is the most widely applicable and convenient method for fine structure analysis in yeast, but suffers the disadvantage that suitable X-ray machines or gamma ray sources are very expensive. Although many other recombinogens are known, none gives a linear dose-response like X-rays and few are as convenient or give as reproducible results. Experiments with Saccharomyces cerevisiae reported in this paper show, however, that the near-ultraviolet radiation emitted by fluorescent sunlamps gives linear dose-response relations, as reproducible results as ionizing radiations, and map distances which correlate highly with those obtained by using (60)Co gamma rays. It is suggested that this convenient recombinogen may be a suitable low-cost substitute for ionizing radiations in fine structure mapping.     

Fungal Catabolism of Crown Gall Opines

This study was conducted to determine the capacities of 37 fungi to utilize various crown gall opines as their sole carbon and nitrogen source. One strain of Fusarium solani, two of Cylindrocarpon destructans, and six of Cylindrocarpon heteronema catabolized octopine, mannopine, octopinic acid, succinamopine, or a combination of these opines. One C. heteronema and one Fusarium dimerum strain grew only on succinamopine. None of the fungal isolates had the ability to grow on nopaline. The catabolism of opines by fungi was confirmed by the disappearance of the opine from the growth medium and by an increase in final mycelial dry weight with rising initial concentration of test substrate. This study thus shows that the catabolism of opines is not restricted to bacteria.     

Use of activated clotting time for monitoring anticoagulation during cardiopulmonary bypass in infants and children with congenital heart disease

The use of a fixed dosage schedule was compared with the use of activated clotting time (ACT) for determining heparin and protamine dosages during and after cardiopulmonary bypass disease. Use of the ACT resulted in a statistically significant increase in heparin dosage and a statistically significant reduction of postoperative blood loss. With ACT use, chest tubes were retained for a shorter period of time, and the incidence of serious postoperative hemorrhage was reduced from 44% to 18%. These results confirm the superiority of the ACT method for monitoring intraoperative anticoagulation in pediatric patients with congenital heart disease.