Characterization of the human jumonji gene

While constructing a cDNA library of human embryos, we have isolated a clone homologous to jumonji, a mouse gene required for neural tube formation. We have determined the complete coding sequence of the human homologue (JMJ) and deduced the amino acid sequence of the putative protein. We show here that human and mouse jumonji putative proteins are homologous and present 90% identity. During human embryogenesis, JMJ mRNAs are predominantly expressed in neurons and particularly in dorsal root ganglion cells. They are also expressed in neurons of human adult cerebral cortex. In view of these observations, we propose JMJ as a candidate gene for developmental defects of the central nervous system in the human. The human JMJ gene maps at position 6p24-6p23.      

Long-term deficits after somatosensory cortical lesions in rats.

Rats having either sham operations or one-stage bilateral lesions of the two somatosensory areas of the cortex were tested for acquisition of five tactile discriminations after postoperative recovery intervals of 14, 35, 180, 365 or 730 days. The group with lesions performed worse than its time-matched control group in every instance, and there was no evidence for recovery of function with the longer postoperative recovery periods. These results suggest that time per se is not a significant determinant of restitution after somatosensory cortical ablations.     

How to use Chlamydia antibody testing in subfertility patients

Screening for tubal factor subfertility by means of Chlamydia antibody testing (CAT) was introduced into the initial work-up of subfertile couples several years ago. The results reported, however, are heterogeneous, and no uniformity exists in cut-off levels of titres, or in definitions of tubal factor subfertility. We performed a prospective cohort study to evaluate the implications of varying the definitions of tubal pathology and of modifying the cut-off levels on the clinical impact of CAT in predicting tubal factor subfertility. In 227 consecutive patients who attended our fertility clinic, the Chlamydia IgG antibody titre was determined and related to tuboperitoneal abnormalities at laparoscopy as a reference standard. According to received operating characteristic (ROC) curve analysis, a titre of 16 is the optimum cut-off level. Increasing the cut-off level improves specificity and positive likelihood ratio (LR+), at the expense of sensitivity and negative LR (LR-). Changing the definition of tubal factor subfertility from unspecified tuboperitoneal abnormalities into extensive adhesions and/or bilateral distal tubal occlusion improves LR+, LR- and kappa significantly. We conclude that CAT is more accurate in predicting severe distal tubal pathology than unspecified tuboperitoneal abnormalities. Although from a statistical point of view a titre of 16 is the optimum cut-off level, from a clinical point of view 32 or 64 may be preferable, depending on the aim of screening and the inception cohort.      

Malignancy may adversely influence the quality and behaviour of oocytes

A case series of eight cycles of in-vitro fertilization (IVF) in five women diagnosed with malignant disorders is presented. These patients chose to defer definitive treatment for a chance for preservation of potential fertility. The response of these patients to ovarian stimulation, and the outcome, was compared with 17 IVF cycles in 12 age- matched patients with isolated tubal infertility. An apparent adverse influence of malignant disease on the quality and behaviour of oocytes was observed. Despite a comparable total number of oocytes per cycle in the two groups, a significantly reduced percentage of mature oocytes was retrieved per cycle from patients with malignant diseases. The oocytes from patients with malignant disorders were of a poorer quality and exhibited a significantly impaired fertilization rate compared to the controls. We propose that neoplastic processes, irrespective of the site or cell of origin, may have a detrimental impact on the biology of oocytes, an effect akin to that seen on spermatozoa in men with certain malignancies.      

Distribution of mutations in the PEX gene in families with X-linked hypophosphataemic rickets (HYP)

Mutations in the PEX gene at Xp22.1 (phosphate-regulating gene with homologies to endopeptidases, on the X-chromosome), are responsible for X-linked hypophosphataemic rickets (HYP). Homology of PEX to the M13 family of Zn2+ metallopeptidases which include neprilysin (NEP) as prototype, has raised important questions regarding PEX function at the molecular level. The aim of this study was to analyse 99 HYP families for PEX gene mutations, and to correlate predicted changes in the protein structure with Zn2+ metallopeptidase gene function. Primers flanking 22 characterised exons were used to amplify DNA by PCR, and SSCP was then used to screen for mutations. Deletions, insertions, nonsense mutations, stop codons and splice mutations occurred in 83% of families screened for in all 22 exons, and 51% of a separate set of families screened in 17 PEX gene exons. Missense mutations in four regions of the gene were informative regarding function, with one mutation in the Zn2+-binding site predicted to alter substrate enzyme interaction and catalysis. Computer analysis of the remaining mutations predicted changes in secondary structure, N-glycosylation, protein phosphorylation and catalytic site molecular structure. The wide range of mutations that align with regions required for protease activity in NEP suggests that PEX also functions as a protease, and may act by processing factor(s) involved in bone mineral metabolism.      

Die sogenannte Impfwut

Zusammenfassung Die im Verlaufe antirabischer Vaccinationen gelegentlich auftretenden paralytischen Komplikationen sind nicht virusbedingt, sondern die Folge einer Antigen-Antikörperreaktion nach Sensibilisierung mit dem organspezifischen Hirnantigen, das in den nicht ätherisierten Tollwutvaccinen enthalten ist. Diesbezüglich von einer Impfwut zu sprechen, erscheint nicht gerechtfertigt. Vielmehr sollte dieser Terminus für die postvaccinalen Komplikationen reserviert bleiben, bei denen der Ausbruch der Erkrankung in direktem Kausalzusammenhang zu dem in einer Vaccine enthaltenen lebenden Virus fixe steht.     

Alternative splicing of exon 14 determines nuclear or cytoplasmic localisation of fmr1 protein isoforms

Impaired expression of the FMR1 gene is responsible for the fragile X mental retardation syndrome. The FMR1 gene encodes a cytoplasmic protein with RNA-binding properties. Its complex alternative splicing leads to several isoforms, whose abundance and specific functions in the cell are not known. We have cloned in expression vectors, cDNAs corresponding to several isoforms. Western blot comparison of the pattern of endogenous FMR1 proteins with these transfected isoforms allowed the tentative identification of the major endogenous isoform as ISO 7 and of a minor band as an isoform lacking exon 14 sequences (ISO 6 or ISO 12), while some other isoforms (ISO 4, ISO 5) were not expressed at detectable levels. Surprisingly, in immunofluorescence studies, the transfected splice variants that exclude exon 14 sequences (and have alternate C-terminal regions) were shown to be nuclear. Such differential localisation was however not seen in subcellular fractionation studies. Analysis of various deletion mutants suggests the presence of a cytoplasmic retention domain encoded in exon 14 and of a nuclear association domain encoded within the first eight exons that appear however to lack a typical nuclear localisation signal.      

Einflu\ vonBordetella pertussis auf das lymphatische Gewebe von Mäusen

Zusammenfassung Verglichen mit phänotypisch normalen Kontrolltieren fanden sich bei NMRI-Mäusen mit kongenitaler Thymusaplasie nur wenige Peyersche Plaques, die zudem nur aus diffusen Lymphocytenansammlungen bestanden. Auch die Milzfollikel waren erheblich verkleinert, wobei die periarteriolären Follikelbereiche gelegentlich zellärmer erschienen. Das wei\e Blutbild der thymuslosen Tiere war durch eine mä\ige Lymphopenie charakterisiert. Obwohl die Milzen der Mäuse mit kongenitaler Thymusaplasie keine Verringerung der Zahlen an präexistenten 19S-Hämolysin-bildenden Zellen aufweisen, waren nach Erstimmunisierung mit 4·10⁸ Schaferythrocyten nur minimale Zahlen an 19S-Produzenten und 7 S-Produzenten nachweisbar. Die sekundäre antigene Stimulierung mit 4·10⁸ Schaferythrocyten führte zu keiner nennenswerten Gedächtnisreaktion, was anzeigt, da\ die Bildung von Gedächtniszellen ein thymusabhängiger Proze\ ist. Die alleinige intraperitoneale Injektion abgetöteter Zellen von B. pertussis führte zu Hypersplenie, Leukozytose und Vermehrung der präexistenten Zahlen an 19S-Hämolysin-bildenden Zellen. Im Gegensatz zu Normaltieren, wo die Leukocytose durch eine Vermehrung von Granulocyten und Lymphocyten charakterisiert ist, fehlte bei den thymuslosen Tieren die Lymphocytenvermehrung. Der Zunahme der Blutgranulocyten entsprach eine verstärkte Granulopoese in Leber und Milz. Bei Normaltieren fand sich nach Erstimmunisierung durch simultane Applikation von 4·10⁸ Schaferythrocyten und 3·10⁹ abgetöteten Zellen vonB. pertussis eine verstärkte und verlängerte Produktion von 19S- und besonders 7S-Hämolysin-bildenden Zellen, verbunden mit einer verstärkten Präparation des lymphoretikulären Gewebes für die anamnestische Reaktion. Bei den Mäusen mit kongenitaler Thymusaplasie beschränkte sich der Adjuvanseffekt allem auf die Frühphase der immunologischen Primärreaktion, was seinen Ausdruck in einer deutlichen Vermehrung der 19S-Produzenten und einer nur geringgradigen Vermehrung der 7S-Produzenten fand.

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Influence ofBordetella pertussis on the lymphatic tissue of miceVIII. The influence ofBordetella pertussis on the primary and secondary immune potential of thymusless (nude) mice

As compared to phenotypically normal controls, in the intestine of NMRI mice with congenital aplasia of the thymus (nude) only small numbers of Peyer's patches could be detected, which consisted of relatively diffuse accumulations of lymphocytes. The splenic follicles were likewise considerably reduced in size, whereby the thymus-dependent areas occasionally appeared to be deficient in lymphocytes. Furthermore, the thymusless mice were found to have blood lymphopenia. Although the spleens of nude mice contained rather enhanced numbers of pre-existing 19S hemolysin forming cells, only small numbers of 19S producers and very poor numbers of 7S hemolysin-forming spleen cells were demonstrable after the primary immunization with 4·10⁸ sheep erythrocytes. Secondary antigenic stimulation with an equal dose of the erythrocyte antigen did not result in a noteworthy secondary immune response. This evidently indicates that the production of memory cells is a thymus-dependent process. A single intraperitoneal injection of 3·10⁹ pertussis organisms led to splenomegaly, blood leukocytosis and augmentation of the pre-existing 19S hemolysin-forming spleen cells. Whilst in normal controls the blood leukocytosis was found to be due to an increase in both lymphocytes and granulocytes, in nude mice the leukocytosis was solely caused by the increase in granulocytes, this being associated with increased granulopoiesis in spleen and liver. The primary immunization of normal controls with 4·10⁸ sheep erythrocytes and 3·10⁹ pertussis organisms led to an increased and prolonged development of both 19S and 7S hemolysin-producing spleen cells. Furthermore, the bacterial adjuvant increased the process of priming for the secondary immune response. Unlike to this, in mice with congenital aplasia of the thymus adjuvancy of pertussis organisms was restricted to the early phase of the primary immune response, as is documented by the distinct augmentation of 19S producers and a poor increase in the numbers of 7S hemolysin-forming spleen cells.

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Diese Arbeit wurde durch die Deutsche Forschungsgemeinschaft gefördert.