Secondary processing of the Plasmodium falciparum merozoite surface protein-1 (MSP1) by a calcium-dependent membrane-bound serine protease: shedding of MSP133 as a noncovalently associated complex with other fragments of the MSP1

Merozoites of the malaria parasite Plasmodium falciparum possess on their surface proteolytically processed fragments of the merozoite surface protein-1 (MSP1). Secondary processing of one of these fragments, MSP1₄₂, always occurs prior to, or at the point of successful erythrocyte reinvasion. It is shown that a product of this secondary processing, MSP1₃₃, is shed in the form of a noncovalently-associated complex with a number of other proteins, including the MSP1-derived species MSP1₃₈ and MSP1₈₃. Secondary processing of MSP1₄₂, is inhibited by the chelating agents ethylenediaminetetraacetic acid (EDTA) and ethyleneglycol-bis-(β-aminoethyl ether)-tetraacetic acid (EGTA), and this inhibition is reversible by addition of excess calcium. Secondary processing occurs in preparations of washed, disrupted merozoites, and is inhibited by the protease inhibitors phenylmethylsulphonyl fluoride (PMSF) and diisopropyl fluorophosphate (DFP), indicating that the protease responsible is a membrane-associated serine protease.     

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.      

ATRX encodes a novel member of the SNF2 family of proteins: mutations point to a common mechanism underlying the ATR-X syndrome

It was shown recently that mutations of the ATRX gene give rise to a severe, X-linked form of syndromal mental retardation associated with alpha thalassaemia (ATR-X syndrome). In this study, we have characterised the full-length cDNA and predicted structure of the ATRX protein. Comparative analysis shows that it is an entirely new member of the SNF2 subgroup of a superfamily of proteins with similar ATPase and helicase domains. ATRX probably acts as a regulator of gene expression. Definition of its genomic structure enabled us to identify four novel splicing defects by screening 52 affected individuals. Correlation between these and previously identified mutations with variations in the ATR-X phenotype provides insights into the pathophysiology of this disease and the normal role of the ATRX protein in vivo.      

A mouse model for infectious mononucleosis

Epstein-Barr virus (EBV) is a ubiquitous human gamma-herpesvirus that establishes life-long latency and is associated with lymphoproliferative disorders and the development of several malignancies. EBV infection is frequently, but not always, associated with the development of a syndrome termed infectious mononucleosis. The recent isolation and characterization of a murine gamma-herpesvirus, MHV-68 (gammaHV-68) has provided the first small animal model for studying immunity and pathogenesis of a gamma-herpesvirus in its natural host. MHV-68 has important biological and genetic similarities with the human gamma-herpesviruses. Following intranasal infection of mice with MHV-68, an acute respiratory infection in the lung develops and is cleared, followed by the establishment of latency. Similar to EBV, MHV-68 latency is largely established in B cells, although other cell types can be latently infected. The establishment of latency correlates with a prominent splenomegaly, polyclonal B cell activation with associated autoantibody production, and CD8+ T cell-dominated peripheral blood lymphocytosis, in many aspects mirroring EBV-induced infectious mononucleosis. There are key differences in the MHV-68- and EBV-induced CD8+ T cell responses however. Whereas the expanded CD8+ T cells associated with EBV-induced mononucleosis are largely the outgrowth of T cells responding to lytic viral epitopes elicited during the acute phase of the response, the CD8+ T cell lymphocytosis associated with MHV-68-induced infectious mononucleosis is dominated by an oligoclonal population of T cells expressing Vbeta4+ T cell receptors that are not reactive to acute viral epitopes. The focus of this article will be to highlight the similarities and differences in the infectious mononucleosis syndrome associated with human and murine gamma-herpesviruses.     

Production of type I (alpha/beta) interferon after virus infection of cloned, alloantigen-sensitized mouse T lymphocytes

Mouse T lymphocytes sensitized to alloantigens were cloned by limiting dilution in the presence of interleukin 2. Clones were tested for surface markers Thy-1, Lyt-1 and Lyt-2, and for cytotoxic function. Production of interferon (IFN) by clones either (a) stimulated with allogeneic cells; (b) activated with concanavalin A (Con A); or (c) infected with Semliki Forest virus or Newcastle disease virus were assayed. All clones produced IFN upon Con A stimulation and most after virus infection. Analysis of the IFN produced by a single clone, using anti-IFN antisera, showed that while Con A stimulation induced production of type II IFN (IFN-gamma), the IFN produced after virus infection was type I IFN (IFN-alpha/beta).     

Preliminary assessment of the Con-Bio laser in vivo

A variety of lasers are available for the treatment of pigmented and vascular lesions. This paper presents our preliminary evaluation of the Q-switched 532 nm Con-Bio laser (Continuum Biomedical, Livermore, CA) in a murine model. Mice were anesthetized with intraperitoneal pentobarbital. Laser impacts (10 nsec, 150 mJ max.) were created on the ears at fluences of 1.5, 2, 3, 4, 5, and 6 J/cm 2. Acute study animals were sacrificed 15 min postinjury. Tissues were fixed in formalin and examined via light microscopy. Chronic study animals were allowed to heal for 3 weeks postoperatively. Parameters analyzed included the presence or absence of blistering, hemorrhage, visual assessment of thermal injury, microscopic evidence of vascular coagulation, or disruption and cosmesis. Acute vascular disruption and coagulation were present in all samples. Healing and cosmesis were good at all fluences tested. Tests of the laser in a prefocused mode produced a tissue cavitation effect with a depth of coagulation of 0.73 +/- 0.44 mm and hemorrhage of 0.68 +/- 0.41 mm. A chronic study of the vascular effect on larger vessels (1-2 mm) was conducted on a rabbit ear. Acute disruption was observed, however, all vessels were recanalized when examined grossly and histologically at 3 weeks. These preliminary results suggest that this new laser may be useful for the treatment of cutaneous vascular lesions. Further studies and clinical trials are warranted.     

Using Medicaid fee-for-service data to develop community health center policy

This article presents an analysis of fee-for-service Medicaid data for King County, Washington. This analysis was conducted using Department of Social and Health Services billing records for patients of the community health centers of Seattle-King County (14 primary care sites), the Seattle-King County Department of Public Health (9 primary care sites), and Harborview Medical Center (a large tertiary facility with a primary care outpatient clinic associated with the University of Washington) from January through June, 1992. The complete billing records of all patients who utilized any one of the 24 sites were made available. These records were used to review utilization patterns and patient costs. The implications for community health centers regarding Medicaid managed care, health care reform, and population-based management are discussed.