The expression patterns of mRNA-encoding stem cell factor, internal stem cell factor and c-kit in the prepubertal and adult porcine ovary

The receptor, c-kit, and its ligand, stem cell factor (SCF), are important regulators of ovarian follicle growth and development. The aim of this study was to identify the sites of expression of mRNA for c-kit and SCF in prepubertal and mature (pregnant and non-pregnant) animals. Ovaries were recovered from prepubertal animals, non-pregnant sows and five sows at approximately 3 months of gestation. Ovine SCF and c-kit DNA were cloned into plasmid vectors to produce RNA probes. Expression of mRNA encoding SCF and c-kit were detected via in situ hybridization. Both mRNA were detected throughout ovaries from all animals. This study provides evidence that the growth-factor complex is required throughout follicle development, and also for continued maintenance of the corpus luteum (CL) in the mature animal. SCF mRNA was localized to the granulosa cell layer and was also extensively expressed in endothelial tissue and throughout the CL. c-kit mRNA was detected in the theca layer, oocytes and also in CL. In conclusion, expression of SCF and c-kit mRNA in granulosa and theca cells, respectively, indicate an important interaction between somatic cells throughout follicle development and that in the mature animal, SCF and c-kit potentially have a role in maintaining progesterone secretion by the CL. The observations of continued expression of SCF and c-kit throughout development suggest that there may be differences in the role of this receptor-ligand complex between large mono- vs. poly ovulatory species, such as the pig.     

Limitations of a simple technique for movement compensation via movement-modified fluence profiles

The delivery of intensity-modulated radiation therapy (IMRT) through the dynamic multileaf collimator (dMLC) technique is well known and recently it has been shown how this can be modified to deliver fluence to a body which is moving in a regular and totally predictable manner. This involves making the leaves 'breath' in tandem with the body motion. This note presents an entirely alternative suggestion in which the leaves do not 'breath' at all but the fluence profile to be mapped to the unbreathing leaf motions is modified so that, when sampled by the specified motion, the actual fluence delivered is close to the desired fluence. Limitations of the concept are discussed.     

Regaining chondrocyte phenotype in thermosensitive gel culture

Chondrocyte tissue engineering continues to be a challenging problem. When chondrocytes are duplicated in vitro, it is imperative to obtain an adequate number of cells of optimal phenotype. A temperature-sensitive polymer gel, a copolymer of poly(N-isopropylacrylamide) and acrylic acid (PNiPAAm-co-Aac), has the ability of gelling at 37 degrees C (the lower critical solution temperature, LCST) or above and liquefying below that temperature (Vernon and Gutowska, Macromol. Symp. 1996;109:155-167). The hypothesis of this study was that chondrocytes could (1) duplicate in the copolymer gel; (2) regain their chondrocyte phenotype; and (3) be easily recovered from the gel by simply lowering the temperature below 37 degrees C. Chondrocytes from adult rabbit scapular cartilage were harvested and cultured in a monolayer culture until confluency (approximately 2 weeks). Next, the cells were harvested and seeded into the copolymer gel and cultured for 2-4 weeks. The phenotype of the cultured cells was then characterized. Two groups of control cultures, monolayer and agarose gel, were used to compare their ability to maintain chondrocyte phenotype. The results showed that chondrocytes isolated from rabbit scapula can re-express chondrocyte phenotype in agarose culture and polymer gel culture but not in monolayer culture. Also, cultured chondrocytes can be easily recovered from polymer gel culture by simply lowering the temperature. This new in vitro method of chondrocyte culture is recommended for chondrocyte propagation and regaining chondrocyte phenotype before cell seeding or transplantation.     

Lassa fever. Effective therapy with ribavirin

In a study of Lassa fever in Sierra Leone, West Africa, we identified two variables associated with a high risk of death, and we evaluated the efficacy of ribavirin and Lassa virus-convalescent plasma for the treatment of Lassa fever. A serum aspartate aminotransferase level greater than or equal to 150 IU per liter at the time of hospital admission was associated with a case-fatality rate of 55 percent (33 of 60). Patients with the same risk factor who were treated for 10 days with intravenous ribavirin, begun within the first 6 days after the onset of fever, had a case-fatality rate of 5 percent (1 of 20) (P = 0.0002 by Fisher's exact test). Patients whose treatment began seven or more days after the onset of fever had a case-fatality rate of 26 percent (11 of 43) (P = 0.01). Viremia with levels greater than or equal to 10(3.6) TCID50 per milliliter on admission was associated with a case-fatality rate of 76 percent (35 of 46). Patients with this risk factor who were treated with intravenous ribavirin within the first six days after onset of fever had a case-fatality rate of 9 percent (1 of 11) (P = 0.006), whereas those treated after seven days or more of illness had a fatality rate of 47 percent (9 of 19) (P = 0.035). Oral ribavirin was also effective in patients at high risk of death. Lassa-convalescent plasma did not significantly reduce mortality in any of the high-risk groups. We conclude that ribavirin is effective in the treatment of Lassa fever and that it should be used at any point in the illness, as well as for postexposure prophylaxis.     

Radioimmunoassay of detirelix ([ N-Ac-D-Nal(2)1,D-p-Cl-Phe2,D-Trp3, D-hArg(Et)6(2),D-Ala10]-luteinizing hormone-releasing hormone) in plasma or serum

A procedure for the radioimmunoassay (RIA) of detirelix in plasma or serum at concentrations as low as 0.15 ng/ml is described. Antiserum was produced by deacetylation of the N-terminus amino groups of detirelix and coupling this analog to bovine serum albumin with a carbodiimide and immunizing rabbits with the resultant conjugate. For RIA, 125I-labeled detirelix was used as the tracer and a double antibody procedure was used to separate the free and bound fractions. No purification of samples was required prior to RIA. Accuracy of the method was assessed by adding known quantities of detirelix to detirelix-free plasma and determining the ratio of measured to added analyte. Linear regression analysis for the concentration range 0.15-150.0 ng/ml yielded a regression equation of y = 0.88 X +1.46 and a correlation coefficient of 0.996. Additional validation was obtained from an in vivo study in which [14C]detirelix was administered to monkeys and plasma clearance profiles were determined by RIA and an HPLC-radiochemical method. The RIA results were in good agreement with those obtained by the HPLC method.     

Relationship of glucocorticoid suppression of arachidonic acid metabolism to alteration of neutrophil function

The role of arachidonic acid metabolism in glucocorticoid-induced suppression of neutrophil function was investigated. In vivo treatment of cattle with dexamethasone decreased the production of lipoxygenase products of arachidonic acid metabolism (leukotriene B4 [LTB4] and H-5-hydroxy-6,8,11,14-eicosatetraenoic acid [5-HETE]) in neutrophils. We previously reported that in vivo dexamethasone treatment resulted in alteration of in vitro neutrophil oxidative metabolism, iodination, antibody-dependent cell-mediated cytotoxicity (ADCC), and random migration. To determine if the decrease in production of LTB4 and 5-HETE were responsible for the changes in neutrophil function, neutrophils were treated in vitro with inhibitors of the lipoxygenase enzyme (BW755c and nordihydroguaiaretic acid), and their function was evaluated. A decrease in neutrophil oxidative metabolism and iodination was observed, but there was no effect on neutrophil random migration or ADCC. The results suggest that in vivo treatment with glucocorticoids inhibit neutrophil oxidative metabolism and iodination by decreasing the formation of lipoxygenase products of arachidonic acid metabolism but alter neutrophil random migration and ADCC by a mechanism independent of arachidonic acid metabolism.     

Frequency and replication status of the fragile X, fra(X)(q27-28), in a pair of monozygotic twins of markedly differing intelligence.

Chromosome analysis using conventional staining, G banding, and, after BUdR incorporation, two R banding methods, one using Hoechst and one acridine orange, were performed on lymphocytes from a pair of female monozygotic twins. The culture conditions were designed to show the presence of the fragile X (q27-28) which had previously been found to be segregating in the family. One twin was of higher than normal intelligence and the other had been diagnosed as mentally retarded. The frequency of the occurrence of the early/active fragile X compared to the overall total of informative fragile X was determined using both methods described above and was also compared with previous published data in the form of a graph showing percentage of early/active fragile X against intelligence.     

The complications of extracorporeal shockwave lithotripsy: management and prevention

Extracorporeal shockwave lithotripsy has been shown to be a safe and effective method of treating most upper urinary tract stones. Major complications, although few, include haemorrhage, septicaemia, "Steinstrasse" formation and cardiac arrhythmias. The experience from 600 consecutive cases is reviewed and methods of prevention are discussed.