Drosophila Cyclin B3 is required for female fertility and is dispensable for mitosis like Cyclin B

Cyclin B3 has been conserved during higher eukaryote evolution as evidenced by its identification in chicken, nematodes, and insects. We demonstrate that Cyclin B3 is present in addition to Cyclins A and B in mitotically proliferating cells and not detectable in endoreduplicating tissues of Drosophila embryos. Cyclin B3 is coimmunoprecipitated with Cdk1(Cdc2) but not with Cdk2(Cdc2c). It is degraded abruptly during mitosis like Cyclins A and B. In contrast to these latter cyclins, which accumulate predominantly in the cytoplasm during interphase, Cyclin B3 is a nuclear protein. Genetic analyses indicate functional redundancies. Double and triple mutant analyses demonstrate that Cyclins A, B, and B3 cooperate to regulate mitosis, but surprisingly single mutants reveal that neither Cyclin B3 nor Cyclin B is required for mitosis. However, both are required for female fertility and Cyclin B also for male fertility.     

Pooling of sera for human immunodeficiency virus (HIV) testing: an economical method for use in developing countries.

To determine whether donated blood samples in African countries could be pooled, then tested for the presence of human immunodeficiency virus (HIV) antibodies with a single test without loss of accuracy, a single test on five pooled samples was used, followed by individual testing of positive pools. This resulted in no loss of either sensitivity or specificity. Pooling 10 samples resulted in a loss of sensitivity for low antibody titre specimens. Pooling reduced the costs of screening by 70% and time needed for analysis. It is concluded that pooling of five samples for HIV screening may result in a substantial reduction in costs; in countries where the prevalence of HIV is higher than the 2-3% found in Zimbabwean donors, however, savings may not be as great.     

Chlamydial pneumonitis induced in newborn guinea pigs.

One- to three-day-old guinea pigs were inoculated intranasally with the chlamydial agent of guinea pig inclusion conjunctivitis. Physical signs of infection included a marked increase in respiration rate on days 5 to 10 of infection and radiographic evidence of pneumonia on day 6. When animals were killed at various times after infection and lung tissue was examined by histopathology, evidence of pneumonia was found beginning on day 4 and lasting as long as day 12, with maximal pathological changes on days 6 to 8. The pneumonia was generally unilateral and consisted of an acute inflammatory component in the bronchioles with granulocytes in both the lumen and the wall of the bronchioles and an interstitial and intra-alveolar mononuclear infiltrate in the parenchyma of the lung. Chlamydial antigen was detected in the bronchial epithelial cells by immunoperoxidase staining, and the guinea pig inclusion conjunctivitis organism was isolated from lung tissue on days 6 to 9. No other significant bacteria were isolated from lung tissue or seen on gram stains of lung sections. Both immunoglobulin M and immunoglobulin G serum antibodies to the guinea pig inclusion conjunctivitis agent were detected as early as day 8 and reached peak levels on day 12. The infection was apparently self-limiting. This model presents the opportunity to investigate pathophysiological and immunological aspects of chlamydial respiratory infections in a neonatal animal.     

Experimental IgA-IgG nephropathy induced by a viral respiratory pathogen. Dependence on antigen form and immune status.

BACKGROUND: The etiology of IgA nephropathy (IgAN), the most common form of glomerulonephritis in the world, remains an enigma. Episodes of nephritis are frequently preceded by acute viral respiratory syndromes, but few experimental models associated with acute viral infection exist. EXPERIMENTAL DESIGN: An animal model of IgAN involving Sendai virus, a rodent parainfluenza virus similar to many human respiratory viruses, is described. Mice were either naturally infected or chronically mucosally immunized with virus. Immunized mice were then challenged intravenously with various physical forms of antigen to simulate viremia or antigenemia secondary to acute viral exposure. Twenty-four hours after challenge of immunized mice or 10 days after natural infection, mice were sacrificed. Anti-viral antibody titers, glomerular immune deposits, and glomerular function were evaluated. RESULTS: Chronic mucosal immunization of mice with Sendai virus resulted in a vigorous serum IgA (and IgG) anti-viral immune response, associated with comparable degrees of IgA, IgG, IgM, and antigen deposits in the glomeruli of both challenged and unchallenged mice. Only immunized, challenged mice developed significant proteinuria and hematuria. Neither deposits nor glomerular dysfunction was seen in controls. The physical form of antigen was important for altered glomerular function; although immunized mice challenged with either live or dead virions had a high incidence of hematuria, mice challenged with purified viral glycoproteins did not, even though all mice exhibited comparable immune deposits. Significant deposits of C3 were not present and were not required for glomerular injury. Finally, naturally infected mice exhibited a milder form of IgAN without hematuria. CONCLUSIONS: The experimental conditions for acute exposure to a natural viral respiratory pathogen of mice leading to IgAN are described. This model may be useful both to probe infection-related IgAN, and to facilitate the understanding of the various mechanisms responsible for IgAN.     

Prevotella bivia as an unusual cause of endocarditis

A case of monomicrobial endocarditis due toPrevotella bivia in a 60-year-old man without previous cardiac lesions is reported. The extremely indolent course with multiple systemic emboli as the only clinical manifestation occurring at least seven months before diagnosis and the persistently negative blood cultures were remarkable features of this case. The incidence, clinical characteristics, treatment and outcome of published cases of infective endocarditis due to anaerobic bacteria are briefly reviewed.     

Production of anti-NC1 antibody by affected male dogs with X-linked hereditary nephritis: A probe for assessing the NC1 domain of collagen type IV in dogs and humans with hereditary nephritis

Summary Some patients with hereditary nephritis (HN) who have received a renal transplant have been shown to form antibody with specificity for the NC1 domain of collagen type IV, a major constituent of glomerular basement membranes (GBM). We attempted to duplicate this phenomenon in a family of dogs with X-linked HN, a model for human X-linked HN, by immunizing affected male dogs with normal dog NC1 domain. A collagenase digest was prepared from normal dog GBM, the NC1 domain was separated into dimer (50 kDa) and monomer (24 kDa and 26 kDa) components by SDS-PAGE, and injected into two affected male dogs. Antisera obtained from both dogs contained antibody which reacted with the NC1 domain of dog and human GBM by a plate-binding radioimmunoassay, bound to the dimer and 26 kDa monomer bands by Western blotting, and staining dog and human GBM by immunofluorescence (IF). The affected male dog antiserum reacted equally by radioimmunoassay with the NC1 domain isolated from GBM of unaffected, affected male, and carrier female dogs in the family with X-linked HN, and bound by Western blotting to dimers and the 26 kDa monomer band of the NC1 domain of GBM in each group of dogs. However, the affected male dog antiserum differentiated these dogs by IF; it produced global staining of GBM of unaffected dogs, failed to stain GBM of affected male dogs, and produced segmental staining of GBM of carrier female dogs. Absorption of the affected male dog antiserum with normal dog NC1 domain eliminated the staining of dog GBM by IF, whereas staining persisted after absorption with affected male dog NC1 domain. The abnormal staining patterns of GBM seen by IF in the affected male and carrier female dogs and the results of the absorption studies imply an abnormality of one or more determinants in the 26 kDa monomer band of the NC1 domain of their GBM. Amino acid sequencing of this band identified the 1(IV) chain of collagen type IV, a finding that has implications for the pathogenesis of canine X-linked HN. Absent and segmental staining respectively were also seen by IF in GBM of a male and female patient with HN, using the affected male dog antiserum. Thus, the results obtained in affected male and carrier female dogs with X-linked HN may also be relevant to patients with this disease.     

Chromosome anomalies as predictors of recurrence risk for spontaneous abortion

Reproductive histories and chromosomes of spontaneous abortions were studied by segregation analysis in 1890 sibships ascertained through a cytogenetically studied abortion. Normal karyotypes are associated with recurrent abortion. Among abnormal karyotypes, trisomy has an elevated recurrence risk even after adjustment through a liability indicator for maternal age. Possible mechanisms and conflicting evidence in the literature on trisomy are discussed. None of these differences in recurrence risk is large enough to play a significant numerical role in genetic counseling.     

Heteroplasmic Segregation Associated with Trisomy-9 in Cultured Human Cells

In cybrid cells carrying the mitochondrial A3243G MELAS mutation, which were also heteroplasmic for the G12300A suppressor mutation, we observed a transient episode of heteroplasmic instability, resulting in a wide diversification in G12300A heteroplasmy levels and a shift in the average heteroplasmy level from 11 to 29%. These cells were found to be trisomic for chromosome 9, whereas a minority of cells that retained disomy-9 showed no instability. Coculture experiments implied that trisomy-9 cells exhibited a significant growth advantage, but neither heteroplasmy levels, respiratory phenotype nor trisomy-9 itself had direct selective value under standard culture conditions. Mitochondrial nucleoid number was the same (50–100) in cells that had or had not experienced transient heteroplasmic instability, but 1–2 orders of magnitude less than the segregation number in such cells. These findings support the idea that mtDNA partition is under nuclear genetic control, and implicate a locus on chromosome 9 in this regulation.     

Standard atlas space for C57BL/6J neonatal mouse brain

A standard atlas space with stereotaxic co-ordinates for the postnatal day 0 (P0) C57BL/6J mouse brain was constructed from the average of eight individual co-registered MR image volumes. Accuracy of registration and morphometric variations in structures between subjects were analyzed statistically. We also applied this atlas coordinate system to data acquired using different imaging protocols as well as to a high-resolution histological atlas obtained from separate animals. Mapping accuracy in the atlas space was examined to determine the applicability of this atlas framework. The results show that the atlas space defined here provides a stable framework for image registration for P0 normal mouse brains. With an appropriate feature-based co-registration strategy, the probability atlas can also provide an accurate anatomical map for images acquired using invasive imaging methods. The atlas templates and the probability map of the anatomical labels are available at .