Mouse homologues of the human AZF candidate gene RBM are expressed in spermatogonia and spermatids, and map to a Y chromosome deletion interval associated with a high incidence of sperm abnormalities

An RNA-binding motif (RBM) gene family has been identified on the human Y chromosome that maps to the same deletion interval as the 'azoospermia factor' (AZF). We have identified the homologous gene family (Rbm) on the mouse Y with a view to investigating the proposal that this gene family plays a role in spermatogenesis. At least 25 and probably >50 copies of Rbm are present on the mouse Y chromosome short arm located between Sry and the centromere. As in the human, a role in spermatogenesis is indicated by a germ cell-specific pattern of expression in the testis, but there are distinct differences in the pattern of expression between the two species. Mice carrying the deletion Yd1, that maps to the proximal Y short arm, are female due to a position effect resulting in non-expression of Sry ; sex-reversing such mice with an Sry transgene produces males with a high incidence of abnormal sperm, making this the third deletion interval on the mouse Y that affects some aspect of spermatogenesis. Most of the copies of Rbm map to this deletion interval, and the Yd1males have markedly reduced Rbm expression, suggesting that RBM deficiency may be responsible for, or contribute to, the abnormal sperm development. In man, deletion of the functional copies of RBM is associated with meiotic arrest rather than sperm anomalies; however, the different effects of deletion are consistent with the differences in expression between the two species.      

Interchromosomal duplications of the adrenoleukodystrophy locus: a phenomenon of pericentromeric plasticity

A 9.7 kb segment encompassing exons 7-10 of the adrenoleukodystrophy (ALD) locus of the X chromosome has duplicated to specific locations near the pericentromeric regions of human chromosomes 2p11,10p11, 16p11 and 22q11. Comparative sequence analysis reveals 92-96% nucleotide identity, indicating that the autosomal ALD paralogs arose relatively recently during the course of higher primate evolution (5-10 million years ago). Analysis of sequences flanking the duplication region identifies the presence of an unusual GCTTTTTGC repeat which may be a sequence-specific integration site for the process of pericentromeric- directed transposition. The breakpoint sequence and phylogenetic analysis predict a two-step transposition model, in which a duplication from Xq28 to pericentromeric 2p11 occurred once, followed by a rapid distribution of a larger duplicon cassette among the pericentromeric regions. In addition to facilitating more effective mutation detection among ALD patients, these findings provide further insight into the molecular basis underlying a pericentromeric-directed mechanism for non- homologous interchromosomal exchange.      

Cryopreservation of all prezygotes in patients at risk of severe hyperstimulation does not eliminate the syndrome, but the chances of pregnancy are excellent with subsequent frozen-thaw transfers

In-vitro fertilization patients (n = 15) at risk of ovarian hyperstimulation syndrome (OHSS) (oestradiol > or =4500 pg/ml on the day of human chorionic gonadotrophin administration and 25 or more follicles of intermediate or large size) underwent aspiration of all follicles and cryopreservation of all fertilized oocytes at the pronuclear stage. Patients were monitored for up to 2 weeks post- retrieval. Subsequent transfer of cryopreserved-thawed embryos was performed in programmed cycles using exogenous oestrogen and progesterone for endometrial preparation. Two patients (13%) developed OHSS necessitating hospitalization and vaginal aspiration of ascitic fluid. Two other patients (13%) developed moderate OHSS requiring ascitic fluid vaginal aspiration in the office setting, with dramatic improvement of the condition. Subsequent transfer of cryopreserved- thawed embryos yielded a clinical pregnancy rate of 58% per transfer and ongoing or delivery rates of 42 and 67% per transfer and per patient respectively. By eliminating pregnancy potential with cryopreservation of all prezygotes and examining the pregnancy potential with subsequent cryopreserved-thawed transfers, it is concluded that OHSS is reduced, but not eliminated for patients at risk. Subsequent transfer of cryopreserved-thawed prezygotes in a programmed cycle with exogenous steroids yields an excellent pregnancy rate.      

Cell cultures for diagnosis of arbovirus infections in livestock and wildlife

Summary Arboviruses can be isolated in serially propagated cells derived from various vertebrates and invertebrates. Cell cultures can be used for direct detection of antigen by fluorescent antibody and enzyme-linked immunosorbent assays, for nucleic acid hybridization, and for visualization of viruses with electron microscopy. Reagents for enzyme-linked immunosorbent assays for IgM and IgG antibodies, hemagglutination-inhibition, complement fixation, and serum dilution-plaque reduction neutralization tests can be prepared in cell cultures infected with these viruses. Thus, cell cultures can be used as laboratory hosts for essentially all isolation, identification, and serodiagnostic procedures for arboviruses. This paper outlines current methods for diagnosis of arbovirus infections in livestock and wildlife, describes certain of these techniques, and provides references for others.     

An Investigation of Methods of Isolation of β2M1 Globulin (Syn.: Iota Protein, 19S γ Globulin, γ1 Macroglobulin, β2M Globulin) and its Association with Isoagglutinin Activity, together with Preliminary Observations on Other Macroglobulins of Slow Electrophoretic Mobility in Normal Human Serum

An investigation is described of methods of isolation and purification of the high molecular weight protein (here called β_2M1 globulin) which is often associated with the isoagglutinin activity of normal human serum. In the course of these experiments it was observed that ultracentrifugal concentration of this protein from serum fractions gives rise to the simultaneous concentration of two other macroglobulins (here called β_2M2 and β_2M3 globulins).

It was found that the concentration of β_2M1 globulin in normal serum is about 25–50 mg. per 100 ml. In serum from the blood of normal donors of groups A, B and O, isohaemagglutinin activity is associated with the β_2M1 globulin but probably accounts for 1 per cent or less of the total β_2M1-globulin concentration in serum. This activity may represent the so-called `natural' isohaemagglutinin.

The immunological relation is discussed of the normal β_2M1 globulin to certain other members of the `family' of immune globulins (γ globulin and β_2A globulin) and to the pathological macroglobulins occurring in Waldenström's syndrome.

Although rabbit antibody to β_2M1 globulin cross-reacts with γ globulin, suggesting that a portion of each molecule bears antigenic groupings in common, it was found that a considerable degree of β_2M1-globulin specificity could be demonstrated after absorbing antisera with purified γ globulin. Attention is also drawn to the evidence that deficiency of either β_2M1 globulin or γ globulin can occur independently, suggesting different cellular sources of origin.

The pathological macroglobulin demonstrable in Waldenström's syndrome is closely related immunologically to normal β_2M1 globulin but is often deficient in isoagglutinin activity. This immunological relation suggests the use of specific anti-β_2M1-globulin antiserum as a simple means of distinguishing macroglobulinaemia from myelomatosis or other conditions with raised levels of γ globulin.

The β_2M2 and β_2M3 globulins are only demonstrable in ultracentrifugal concentrates of normal serum. Evidence is presented to suggest that these proteins are nevertheless present as trace components of normal serum. The solubility and electrophoretic characteristics of these proteins resemble those of β_2M1 globulin, but they can be distinguished by immunological methods from β_2M1 globulin and each other. Attention is drawn to the evidence that β_2M2 and β_2M3 globulins may also be increased in the serum of some cases of macroglobulinaemia.

     

Cyt1A from Bacillus thuringiensis restores toxicity of Bacillus sphaericus against resistant Culex quinquefasciatus (Diptera: Culicidae)

The 2362 strain of Bacillus sphaericus, which produces a binary toxin highly active against Culex mosquitoes, has been developed recently as a commercial larvicide. It is being used currently in operational mosquito control programs in several countries including Brazil, France, India, and the United States. Laboratory studies have shown that mosquitoes can develop resistance to B. sphaericus, and low levels of resistance have already been reported in field populations in Brazil, France, and India. To develop tools for resistance management, the Cyt1A protein of Bacillus thuringiensis subsp. israelensis De Barjac was evaluated for its ability to suppress resistance to B. sphaericus in a highly resistant population of Culex quinquefasciatus Say. A combination of B. sphaericus 2362 in a 10:1 ratio with a strain of B. thuringiensis subsp. israelensis that only produces Cyt1A reduced resistance by >30,000-fold. Resistance was suppressed completely when B. sphaericus was combined with purified Cyt1A crystals in a 10:1 ratio. Synergism was observed between the Cyt1A toxin and B. sphaericus against the resistant mosquito population and accounted for the marked reduction in resistance. However, no synergism was observed between the toxins against a nonresistant mosquito population. These results indicate that Cyt1A could be useful for managing resistance to B. sphaericus 2362 in Culex populations, and also provide additional evidence that Cyt1A may synergize toxicity by enhancing the binding to and insertion of toxins into the mosquito microvillar membrane.     

Primary gastric T-cell lymphoma of suppressor-cytotoxic (CD8+) phenotype: discordant expression of T-cell receptor subunit beta F1, CD7, and CD3 antigens.

Primary gastric T-cell lymphomas are rare neoplasms, and all but one of the previously phenotyped cases have shown a helper-inducer phenotype. The present case is the second reported case of a primary gastric T-cell lymphoma of suppressor-cytotoxic phenotype. The tumor histology was similar to that described in some forms of node-based peripheral T-cell lymphomas. Phenotypic analysis revealed low expression of pan-T marker CD7, reduced expression of CD3, but higher density and frequency of expression of CD8 antigens that could be predicted on the basis of the pan-T markers. Natural killer cell (NK) related markers CD16, HNK-1 and NKH-1 were not expressed by the neoplastic cells. T-cell receptor (TCR) beta subunit expression was detected on fewer cells than would have been predicted on the basis of CD3 and CD8 expression, and TCR delta chain expression was undetectable.     

Local fetal signal is not required for maintaining IGFBP gene expression in the human decidua: evidence from extrauterine pregnancies

Insulin-like growth factor-II (IGF-II) from the invading extravillous cytotrophoblasts (EVTs) and insulin-like growth factor binding proteins (IGFBPs) from the maternal decidua interact at the feto-maternal interface and regulate implantation and placentation. To determine whether a local stimulus from the fetus is important in the regulation of IGFBP gene expression in the human decidua, we compared the expression of IGFBP genes in intra- and extrauterine (tubal) pregnancies. The expression of IGF-II and IGFBP-1 to IGFBP-6 mRNAs was determined by in-situ hybridization in the Fallopian tubes of extrauterine pregnancies and concurrent decidua (n = 6), and in the placentae and Fallopian tubes of intrauterine pregnancies (n = 6). All six IGFBP mRNAs were identified in the decidualized endometrium and decidualized Fallopian tubes of intra- and extrauterine pregnancies, with IGFBP-1 mRNA being the predominant mRNA. IGFBP-4 was the second most predominant mRNA and was slightly more abundant in the decidua of extrauterine pregnancies than of intrauterine pregnancies. IGF-II mRNA was expressed mainly in cells of fetal origin. The fact that the IGFBP mRNAs were expressed similarly in both intra- and extrauterine pregnancies indicates that the local physical stimulus from an implanting fetus is not necessary to induce or maintain decidual IGFBP gene expression.     

Identification of a critical period for motor development in neonatal rats.

Manipulation of the developing nervous system has provided valuable insights into nervous system function. One important concept to arise from this type of study has been the identification of specific "critical periods" for the development of various functions. A critical period has been most clearly shown for the visual system where monocular eye closure for a few weeks led to functionally significant changes in visually guided behaviors and the connectivity of the visual cortex. Critical periods have also been defined for other sensory systems. Although studies of the effect of manipulating sensory systems during development are sometimes difficult to interpret (e.g. Ref. 7), this difficulty is compounded in the case of the motor system. Problems arise because manipulations of the postnatal motor system are difficult to implement and usually require invasive procedures such as tenotomy, neurotomy, and nerve crush (for review, see Ref. 17). We have approached the problem of manipulating the motor environment by adapting a paradigm widely used to study the experimental effects of simulated weightlessness in adult rats: namely, tail suspension. This method has several advantages for manipulating the motor system: (i) because it is noninvasive, it is less discomforting than neurotomy, tenotomy or nerve crush; (ii) it does not immobilize the animals, they move about the cage and extend and flex their hindlimbs; and (iii) it specifically examines the importance of load-bearing on the development of antigravity muscles and their neuronal circuits.(ABSTRACT TRUNCATED AT 250 WORDS)