Sindbis virus (SINV) is the prototype member of the Alphavirus genus, whose members cause severe human diseases for which there is no specific treatment. To ascertain host factors important in the replication of the SINV RNA genome, we generated a SINV expressing nsP4, the viral RNA-dependent RNA polymerase, with an in-frame 3؋Flag epitope tag. Proteomic analysis of nsP4-containing complexes isolated from cells infected with the tagged virus revealed 29 associated host proteins. Of these, 10 proteins were associated only at a later time of infection (12 h), 14 were associated both early and late, and five were isolated only at the earlier time (6 h postinfection). These results demonstrate the dynamic nature of the virus-host interaction that occurs over the course of infection and suggest that different host proteins may be required for the multiple functions carried out by nsP4. Two related proteins found in association with nsP4 at both times of infection, GTPase-activating protein (SH3 domain) binding protein 1 (G3BP1) and G3BP2 were also previously identified as associated with SINV nsP2 and nsP3. We demonstrate a likely overlapping role for these host factors in limiting SINV replication events. The present study also identifies 10 host factors associated with nsP4 6 h after infection that were not found to be associated with nsP2 or nsP3. These factors are candidates for playing important roles in the RNA replication process. Identifying host factors essential for replication should lead to new strategies to interrupt alphavirus replication.Alphaviruses are important human and animal pathogens, causing fever, rash, arthritis, encephalitis, and death (reviewed in reference 18). These enveloped positive-strand RNA viruses cycle in nature through small vertebrate reservoir hosts and invertebrate vectors, most commonly mosquitoes. Infection of larger mammals, including humans, through the bite of an infected mosquito can lead to severe disease, for which there is no specific therapy. Viruses in the Alphavirus genus share common genomic organization and replication strategies, and much has been learned through study of Sindbis virus (SINV), the prototype virus in this genus. SINV, like the other alphaviruses, enters cells by receptor-mediated endocytosis and after fusion in the endosome releases the positive-strand RNA genome, which is translated in the cytoplasm to generate a nonstructural polyprotein, P123 (reviewed in reference 36). Translational readthrough of an opal termination codon also results in the production of the polyprotein P1234. The activity of a cysteine protease residing in nsP2 mediates polyprotein processing, which results in the generation of P123 and nonstructural protein 4 (nsP4), the viral RNA-dependent RNA polymerase. Together, P123 and nsP4 produce a full-length negative-strand copy of the genomic RNA. Further regulated sequential processing of the polyprotein results in replication complexes active in both positive and negative strand RNA synthesis. After complete proteolytic proces...

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1. A Klinefelter patient is reported with the sex chromosome constitution XXYY, and the modal number of 48 chromosomes. 2. This case excludes the possibility of maternal non-disjunction as a cause. 3. The evidence of paternal and maternal errors in Klinefelter's and Turner's syndromes is discussed, and it seems that paternal errors are more common than maternal. 4. The cause of Barr body formation is discussed with preference for the theory of paternal origin of heteropycnotic sex chromosomes. This theory may explain the existence of chromatin positive Turner's and chromatin negative Klinefelter's. 5. The Y-chromosome is considered in connection with the postulated XYY-male and with the non-mosaic true hermaphrodite. 6. A simple definition of the Klinefelter syndrome is offered.

Over the past 2 decades, hematopoietic stem cell transplantation (HSCT) has been used as therapy for selected inherited metabolic and genetic diseases (IMGDs). The primary objective of HSCT for these disorders has been to promote long-term survival, optimize quality of life, and improve neurocognitive performance. We performed 45 HSCTs for 44 children with IMGDs (13 related and 32 unrelated); 24 HSCTs for 23 children with Hurler syndrome, 8 for malignant infantile osteopetrosis, 6 for X-linked adrenoleukodystrophy, 2 for metachromatic leukodystrophy, 2 for Gaucher disease, 1 for Ganglioside Monosialic Acid (GM) gangliosidosis, 1 for sialiosis (type 2), and 1 HSCT for Niemann-Pick type A. At a median follow-up of 7.2 years (range: 2.2 to 17.6 y) 18 of 23 patients with Hurler syndrome are alive, 15 attended regular school. Thirteen of 18 were ambulatory, 2 had mobility difficulties, and 1 uses wheelchair. For non-Hurler patients, 5 children suffered secondary graft failure and 4 of them died from progressive disease. The remaining children with osteopetrosis are alive and most children attended regular school. One out of the 4 survivors with adrenoleukodystrophy has been transferred to the adult follow-up clinic and he is in full-time employment. Parents' perspectives and expectations of HSCT in these IMGDs were positive and supportive to continue to offer HSCT for these disorders.