Evidence for regulation of cartilage differentiation by the homeobox gene Hoxc-8

Homeobox genes of the Hox class are required for proper patterning of skeletal elements, but how they regulate the differentiation of specific tissues is unclear. We show here that overexpression of a Hoxc-8 transgene causes cartilage defects whose severity depends on transgene dosage. The abnormal cartilage is characterized by an accumulation of proliferating chondrocytes and reduced maturation. Since Hoxc-8 is normally expressed in chondrocytes, these results suggest that Hoxc-8 continues to regulate skeletal development well beyond pattern formation in a tissue-specific manner, presumably by controlling the progression of cells along the chondrocyte differentiation pathway. The comparison to Hoxd-4 and Isl-1 indicates that this role in chondrogenesis is specific to proteins of the Hox class. Their capacity for regulation of cartilage differentiation suggests that Hox genes could also be involved in human chondrodysplasias or other cartilage disorders.     

Endothelium-derived relaxing factor activates calcium-activated potassium channels of resistance vessel smooth muscle cells

Direct observation was made by using the patch-clamp technique with a specially designed microperfusion system to investigate the effect of acetylcholine (Ach 10(-6) mol/L) elicited endothelium-derived relaxing factor (EDRF) on the calcium-activated potassium channel (IK(Ca)) in the smooth muscle cells of mesenteric resistance vessels in Wistar rats. Activation of IK(Ca) was firstly observed by inducing the elicited EDRF or sodium nitroprusside (SNP 10(-8) mol/L) under various clamping voltages in cell-attached configuration. While the pipette solution contained KCl 126 mmol/L and the bath solution contained KCl 5.9 mmol/L, two types of conductances of calcium-activated potassium current being 76.4 +/- 2.3 pS (mean +/- S.E. n = 7) and 160.3 +/- 7.5 pS (mean +/- S.E. n = 7) were recorded during the EDRF activation, one type of conductance being 100.5 +/- 2.8 pS (mean +/- S.E. n = 6) was activated by nitric oxide (NO) which is an effective component from SNP. Differences in kinetic characteristics of these channels between EDRF and NO activation were found, particularly the probability of the channel being open in EDRF activation was obviously greater than that in NO stimulation. It has been shown that the potassium channel mechanisms involved in the EDRF and NO actions might be different.     

Primary ciliary dyskinesia: ultrastructural defects and clinical features

Golden hamsters were exposed to conditions of 2.5 times normal gravity (hypergravity, HG) for 4 months. During this period, tests were carried out to study equilibrium maintenance, swimming behaviour and open-field behaviour of these HG hamsters and of control hamsters living in a normal-gravity environment. The tests proved to be useful devices for detecting differences in perceptive-motor behaviour between HG hamsters and control hamsters. The HG hamsters had more difficulties in balancing on tubes and orientation during swimming. In the open-field study, the HG hamsters showed less locomotor activity than control hamsters. However, no differences were observed between the groups in washing, rearing and number of times having defaecation. These findings indicate that the daily transition from 2.5 to 1 g was not experienced as stressful by the hamsters, although performance on several perceptive-motor tasks was decreased, especially during the first weeks.     

Extended life of human corneal endothelial cells transfected with the SV40 large T antigen

PURPOSE: To transfect human corneal endothelial cells with a plasmid vector coding for the SV40 large T antigen to extend the life of the cells in culture. METHODS: Human corneal endothelial cells were transfected with the SV40 large T antigen-coding plasmid pSV3neo using the electroporation method. Transfected and control cells were propagated in culture until senescence. Polymerase chain reaction and immunofluorescence were used to demonstrate messenger RNA and protein, respectively, for the Simian virus 40 large T antigen in the transfected cells. Polymerase chain reaction and hot blotting were used to demonstrate messenger RNA coding for several growth factors and receptors in transfected and control cells. RESULTS: The transfected cells continued to proliferate to 38 passages (more than 120 population doublings) in culture (control cells, 8 population doublings). Transfected cells, but not control cells, expressed messenger RNA coding for the Simian virus 40 large T antigen. Similarly, immunofluorescent staining with monoclonal antibodies demonstrated that the Simian virus 40 large T antigen protein was present in the nucleus of the transfected cells. Transfected cells were shown to produce messenger RNA coding for epidermal growth factor, epidermal growth factor receptor, basic fibroblast growth factor, fibroblast growth factor receptor-1, interleukin-1 alpha, the interleukin-1 receptor, transforming growth factor beta-1, and the glucocorticoid receptor. Qualitative expression of the messenger RNA coding for each of these modulators was similar in proliferating primary corneal endothelial cells and proliferating or confluent transfected corneal endothelial cells. CONCLUSIONS: In culture, the life of human corneal endothelial cells transfected with a plasmid vector coding for the Simian virus 40 large T antigen is extended. This study suggests that human corneal endothelial cells have the capacity for extensive proliferation, but the proliferation of untransfected cells is regulated through mechanisms that have not yet been characterized.     

Omentoplasty in the management of filarial lymphoedema

OBJECTIVE: Peer review is an important part of the publication of scientific results. We analysed the peer review process of abstracts submitted to the European Association for the Study of the Liver (EASL) congress in Copenhagen in 1995. DESIGN: Comparative study. All abstracts were reviewed by all six members of the EASL scientific committee. Furthermore, each abstract was evaluated by three to six experts within 17 specific study fields, appointed by the scientific committee. All abstracts received a score on a scale from 1 to 5, one being the best. For each abstract the average was calculated for the scores of the committee members and the expert reviewers separately. MAIN OUTCOME MEASURE: Abstract score. RESULTS: There was a significant difference in the scores between the committee members, who scored 3.31 on average, and the expert reviewers, who scored 3.02 on average. Within most of the study fields the tendency was that the expert reviewers gave the highest scores. Moreover, there was a significant variation in the estimates among the individual reviewers as well as between the 2 groups of reviewers. Only 25% of the abstracts that received the best scores by the expert reviewers were classified in the same way by the committee members. The predictive value of a positive evaluation by the committee members was 0.6, the predictive value of a negative evaluation was 0.90. CONCLUSIONS: There was a considerable variation in the peer review process for abstracts submitted to the 1995 EASL congress depending on selection of reviewers. Various types of peer review strategies should be tested and evaluated to secure the highest possible scientific quality of published research results.     

Cre/loxP-mediated in vivo excision of large segments from yeast genome and their amplification based on the 2microm plasmid-derived system

The loss of spleen may lead to fatal bacterial infections. To prevent this, splenic autotransplantation has been performed in humans and experimental animals. However, there is still controversy about the protective function of this procedure. Since innervation plays an important role in splenic function, we investigated whether splenic regenerates are re-innervated, and whether this depends on the donor and host age. Splenic tissue (30 mg) was implanted into the greater omentum of either young (2 days) or old (12 months) rats, from either young or old syngeneic animals. After 3 months of regeneration, the weight of the regenerates was determined, PGP+ nerve fibers were revealed by immunohistology, and subdivided into nerve fibers of sympathetic (TH+, NPY+) or sensory (SP+, CGRP+) origin. In addition, proliferating (Ki-67 proliferation antigen+) and apoptotic cells (TUNEL technique+) were likewise investigated. No innervation of splenic regenerates was observed after implantation into old hosts, correlating with poorly developed splenic compartments. In contrast, almost normal re-innervation occurred in young hosts after implantation of both young and old splenic tissue. These regenerates showed well-developed splenic compartments and a normal number and tissue distribution of proliferating and apoptotic cells. However, after the implantation of young tissue, the final size of splenic regenerates was three times larger (140 +/- 30 vs. 40 +/- 10 mg). Thus, re-innervation of splenic implants is necessary for their subsequent development. It is determined by host age, whereas the final size of the splenic regenerates is regulated by donor age-dependent factors. This model is useful for studying both the process leading to initial innervation and the consequences of this innervation.