Mutational analysis of the SOX9 gene in campomelic dysplasia and autosomal sex reversal: lack of genotype/phenotype correlations

It has previously been shown that, in the heterozygous state, mutations in the SOX9 gene cause campomelic dysplasia (CD) and the often associated autosomal XY sex reversal. In 12 CD patients, 10 novel mutations and one recurrent mutation were characterized in one SOX9 allele each, and in one case, no mutation was found. Four missense mutations are all located within the high mobility group (HMG) domain. They either reduce or abolish the DNA-binding ability of the mutant SOX9 proteins. Among the five nonsense and three frameshift mutations identified, two leave the C-terminal transactivation (TA) domain encompassing residues 402-509 of SOX9 partly or almost completely intact. When tested in cell transfection experiments, the recurrent nonsense mutation Y440X, found in two patients who survived for four and more than 9 years, respectively, exhibits some residual transactivation ability. In contrast, a frameshift mutation extending the protein by 70 residues at codon 507, found in a patient who died shortly after birth, showed no transactivation. This is apparently due to instability of the mutant SOX9 protein as demonstrated by Western blotting. Amino acid substitutions and nonsense mutations are found in patients with and without XY sex reversal, indicating that sex reversal in CD is subject to variable penetrance. Finally, none of 18 female patients with XY gonadal dysgenesis (Swyer syndrome) showed an altered SOX9 banding pattern in SSCP assays, providing evidence that SOX9 mutations do not usually result in XY sex reversal without skeletal malformations.      

Clinical and familial study of arrhythmogenic right ventricular cardiomyopathy

Objective　To explore the characteristics of arrhythmogenic right ventricular cardiomyopathy (ARVC). Methods　Seven patients with arrhythmogenic right ventricular cardiomyopathy and 34 members of three families were studied. All patients and family members underwent history collection, clinical examination, electrocardiogram (ECG), two-dimensional echocardiography (2-DE) and a signal averaging electrocardiogram. Programmed ventricular stimulation was performed in five patients. Results　All patients and family members had normal morphologic characteristics and normal function of the left ventricular by 2-DE. Fourteen persons had abnormal findings indicating ARVC. Five had enlargement of the right ventricular with diffused hypocontractility, eight had thin and systolic bulging in the focal anterior wall with hypokinesia and one had bulging of the inferior wall. Twenty-five persons (seven patients and 18 family members) had abnormal findings in ECG. Positive ventricular late potential was recorded in 13 persons (six patients). Two to three monomorphic ventricular tachycardia (VT) with left bundle branch block (LBBB) configurations were induced in five patients. Ventricular fibrillation was induced in two patients during the electrophysiologic study (EPS). Five patients had very high pacing threshold and/or ineffective pacing in one or many regions of the right ventricle. Two members of one family died suddenly. One member was a dwarf with ARVC. Spontaneous VT with a left bundle branch block (LBBB) configuration was recorded in five patients, polymorphic VT with extremely short coupling interval in one, and premature ventricular complexes with LBBB configuration in 12 (six patients). Conclusion　Our familial study strongly suggests that ARVC may be a hereditary disease and it is helpful in the diagnosis and detection of ARVC. The most common manifestations were abnormal structure and function of the right ventricle and abnormal ECG of repolarization and ventricular arrhythmia which originates from the right ventricle.     

蝙蝠葛中的新生物碱——蝙蝠葛新林碱

防已科植物蝙蝠葛(Menispermum daurioum DC.)的根茎,即北豆根,是一种重要的中药,具有清热解毒、消肿止痛等功效。已知含有近二十种生物碱,其中五种属双苄基四氢异喹啉类,即蝙蝠葛碱(dauricine,Ⅰ)、蝙蝠葛诺林碱(daurinoline,Ⅱ)、蝙蝠葛新诺林碱     

Role of the lateral and medial hypothalamus in the reproduction of the motoric reaction which is a signal during the development of classical conditioned reflexes

It has been demonstrated in experiments on three dogs that test electrostimulation of the lateral hypothalamus reproduces the motoric reaction which is the signal stimulus during the development of classical alimentary CR (83%) and does not reproduce it during the development of classical defensive CR (93%). The test electrostimulation of the medial divisions of the hypothalamus, like the electrostimulation of the LH, reproduces the signal motoric reaction, but in a smaller percent of cases (62%), during the development of classical alimentary CR, and does not reproduce it during the development of classical defensive CR. The reproduction of the signal motoric reaction during the electrostimulation of the LH is associated with the activation of a backward conditioned connection from the motivational structures of the hypothalamus to the representation of the signal stimulus in the motor cortex.Translated from Zhurnal Vysshei Nervnoi Deyatel'nosti imeni I. P. Pavlova, Vol. 41, No. 1, pp. 41–50, January–February, 1991.     

Retinotectal system of the tortoise, Testudo horsfieldi, Gray (morpho-functional study in the norm and after enucleation).

The optic nerve and mesencephalic optic centre (tectum opticum, TO) of the tortoise, Testudo horsfieldi, Gray, in the norm and after the enucleation have been studies using Golgi's method, electron microscopy and the electrophysiological technique. The optic nerve comprises about 400,000 fibres represented by two classes of axons: myelinated (10%) and unmyelinated (90%). The diameters of the former vary within 0.3-3 mum, of the latter within 0.3-1.1 mum. The neurogramms of the optic nerve contain two components corresponding to the two rates of conduction through different groups of fibres (1.3 musec and 0.5 musec). The stratified synaptic organization of the TO has been investigated. The optic terminals are shown to join in complex synaptic formations at certain levels of the upper layers of the TO (Strata I, II and III). The process of degeneration in the optic system of Testudo is drastically stretched in time. It was demonstrated by the electrophysiological control that the TO potentials induced in response to the nerve stimulation were not registered any longer only 6-6.5 months after the enucleation. Certain groups of retinal fibres of the optic nerve and their terminals in the TO are shown to degenerate differently and asynchronously. Earlier unknown types of destructive changes of the terminals (vesicular, neurofilamentous, glycogen and others) are described. It is suggested that the different types of degeneration of the optic fibres and their terminals are the result of possible biochemical heterogeneity of the retinal ganglious cells. The comparison of the terms and types of degeneration of different groups of nervous fibres in the optic nerve and their terminals in the TO has revealed some correlations suggesting that the myelinated optic fibres produce nervous terminals degenerating by the "dark" type, whereas the unmyelinated optic axons may be subdivided into several groups, each producing nervous terminals subjected to a definite type of destruction ("clear", "vesicular", "neurofilamentous" etc.). The localization of certain groups of therminals at certain levels of the cortical plate of the TO confirms the electrophysiological data on the stratified organization of the retino-tectal projections in the TO of lower vertebrates. It has been shown by the comparison of the results obtained with those reported for Emys orbicularis, L. that the two species have marked differences in the morpho-functional characteristics of the retino-tectal system, neuronic composition of the TO, the character of destructive changes of the terminals after the enucleation and the component composition of polysynaptic complexes and glomerules. Probably these differences may come from the peculiarities of the perception and transformation of visual information in the species.     

Identification of differentially expressed genes in aflatoxin B1- treated cultured primary rat hepatocytes and Fischer 344 rats

Aflatoxin B1 (AFB1), a mutagen and hepatocarcinogen in rats and humans, is a contaminant of the human food supply, particularly in parts of Africa and Asia. AFB1-induced changes in gene expression may play a part in the development of the toxic, immunosuppressive and carcinogenic properties of this fungal metabolite. An understanding of the-role of AFB1 in modulating gene regulation should provide insight regarding mechanisms of AFB1-induced carcinogenesis. We used three PCR- based subtractive techniques to identify AFB1-responsive genes in cultured primary rat hepatocyte RNA: differential display PCR (DD-PCR), representational difference analysis (RDA) and suppression subtractive hybridization (SSH). Each of the three techniques identified AFB1- responsive genes, although no individual cDNA was isolated by more than one technique. Nine cDNAs isolated using DD-PCR, RDA or SSH were found to represent eight genes that are differentially expressed as a result of AFB1 exposure. Genes whose mRNA levels were increased in cultured primary rat hepatocytes after AFB1 treatment were corticosteroid binding globulin (CBG), cytochrome P450 4F1 (CYP4F1), alpha-2 microglobulin, C4b-binding protein (C4BP), serum amyloid A-2 and glutathione S-transferase Yb2 (GST). Transferrin and a small CYP3A-like cDNA had reduced mRNA levels after AFB1 exposure. Full-length CYP3A mRNA levels were increased. When liver RNA from AFB1-treated male F344 rats was evaluated for transferrin, CBG, GST, CYP3A and CYP4F1 expression, a decrease in transferrin mRNA and an increase in CBG, GST, CYP3A and CYP4F1 mRNA levels was also seen. Analysis of the potential function of these genes in maintaining cellular homeostasis suggests that their differential expression could contribute to the toxicity associated with AFB1 exposure.