Ontogenesis of laterality in 3- to 10-yr.-old children: increased unimanual independence grounded on improved bimanual motor function.

Underlying sensorimotor factors, such as intermanual coupling, contributing to optimal laterality, long remain immature. Using the handmotor laterality blackboard, developmental change in symmetric bimanual motor function for 413 children in groups of 3 to 10 years of age shows synchronicity increased between groups 3 and 5 years of age; but between groups 5 and 7 years of age, given immature coupling, one hand may still disturb the movements of the other one. Between groups 7 and 9 years of age, the hands gradually stop disturbing each other and move independently and fluently. Changes in intermanual coupling with increasing unimanual independence represent expression of changing interhemispheric integration across groups. This promotes optimal laterality and task distribution between the hands. Maturation of the corpus callosum is inferred to be a factor in these ontogenetic changes which ultimately lead to optimal left hemisphere specialization for actions such as fluent writing.

Ontogenetic trends in gnostic hand function in 3- to 12-yr-old children.

The intimate relation between the sensory and motor functions of the hands during object manipulation and exploratory touch, the well-known improvement in object handling and constructive performance in ontogenesis and the emergent laterality thereof, assume changes in morphognostic capabilities in children. In this study we tried to corroborate the hypothesis of Mesker that mature and lateralized finger-thumb opposition is preceded by a stage of two-sided manual form agnosia in preschool children, followed by acquisition of morphognosis of the fingers and, finally, the thumbs. This study examined the development of gnostic hand function in 290 children from 3 to 11 years of age who drew the outlines of a meaningless wooden object passively felt with each hand without visual control. Analysis showed a clear ontogenetic change across the two age groups of increasing morphognostic function: 48% of the 6-yr.-olds drew correctly what the fingers of both left and right hands had perceived (thumbs, 14%). Of the 11-yr.-olds 91% and 61% performed perfectly with the right and left hands, respectively. The fingers preceded the thumbs in reproduction by most children, and the correct reproduction by the left thumb precedes that of the right thumb. The ontogenesis of bimanual sensorimotor functioning is discussed in the light of cortical and callosal development.

Two 21-kilodalton components of the Epstein-Barr virus capsid antigen complex and their relationship to ZEBRA-associated protein p21 (ZAP21).

The viral capsid antigen complex of Epstein-Barr virus (EBV), an important serodiagnostic marker of infection with the virus, consists of at least four components, with molecular masses of 150, 110, 40, and 21 kDa. Here we show that the 21-kDa component of the viral capsid antigen consists of products of two EBV genes, BFRF3 and BLRF2. Both products were expressed from late transcripts, were recognized by human antisera, and were present in virions. The BFRF3 product, but not that of BLRF2, fulfilled the definition of ZEBRA-associated protein p21 (ZAP21). In cells in which EBV was lytically replicating, BFRF3 protein was coimmunoprecipitated together with ZEBRA by a rabbit antiserum directed against amino acids 197 to 245 of BZLF1. In EBV-negative cells cotransfected with BZLF1 and BFRF3 expression vectors, BFRF3 was also coimmunoprecipitated with this antiserum. Although this antiserum could not detect BFRF3 on an immunoblot, it was able to immunoprecipitate BFRF3 in the absence of ZEBRA expression. The rabbit antiserum to amino acids 197 to 245 of BZLF1 was found to detect the same epitope at the carboxy end of BFRF3 as was recognized by rabbit antiserum to BFRF3 itself. Thus, coimmunoprecipitation of BFRF3 p21 with ZEBRA appeared to be due to cross-reactivity of the immunoprecipitating antiserum rather than to direct association of ZEBRA and BFRF3 p21.

Localization and diagnostic application of immunodominant domains of the BFRF3-encoded Epstein-Barr virus capsid protein.

The Epstein-Barr virus (EBV) open reading frame BFRF3 encodes a viral structural capsid protein or tegument protein, VCA p18, that is highly immunogenic in humans. In this study, a cluster of immunodominant epitopes in the C-terminus of VCA p18 has been identified. These epitopes were combined into a single synthetic peptide that was analyzed for diagnostic value in an ELISA. This VCA combined peptide appeared to be an excellent VCA marker with remarkable applications for IgG- and IgM-related EBV diagnostics. The combined peptide reacted with 95% of 159 sera that were IgG VCA-positive by immunofluorescence assay. In addition, 95% of sera from 67 persons with confirmed infectious mononucleosis were positive for IgM antibodies to this VCA combined peptide. An IgG-specific seroconversion could be demonstrated by subsequent serum samples during the acute phase of primary EBV infections. Although 97% of the tested sera from nasopharyngeal carcinoma patients had IgG antibodies reacting positively with the VCA combined peptide, only 61% contained IgA anti-VCA combined peptide.

Gene mapping and expression of two immunodominant Epstein-Barr virus capsid proteins.

The genomic localization of two immunodominant genes encoding two proteins of the Epstein-Barr virus capsid antigen (VCA) complex, VCA-p18 and VCA-p40, has been identified. For that purpose, lambda gt11-based cDNA libraries were constructed from HH514.c16 cells induced for virus production. The libraries were screened with a monoclonal antibody, EBV.OT41A, directed against VCA-p40 or with affinity-purified human antibodies against VCA-p18. Sequencing of the inserts of positive plaques showed that VCA-p18 and VCA-p40 are encoded within open reading frames (ORFs) BFRF3 and BdRF1, respectively. Peptide scanning analysis of the predicted protein of ORF BdRF1 resulted in defining the epitope of monoclonal antibody EBV.OT41A at the C-terminal region. The dominant VCA-p18 reactivity of human sera can be completely inhibited by preadsorption with Escherichia coli-expressed BFRF3-beta-galactosidase. Serum of a rabbit immunized with BFRF3-beta galactosidase reacts with a VCA-specific protein of 18 kDa. In addition, BFRF3-beta-galactosidase affinity-purified antibodies react with VCA-p18 of virus-producing cells (HH514.c16). Complete inhibition of viral DNA polymerase activity by phosphonoacetic acid is associated with the absence of RNAs and protein products of both ORFs, indicating that VCA-p18 and VCA-p40 are true late antigens.

Identification and molecular characterization of two diagnostically relevant marker proteins of the Epstein-Barr virus capsid antigen complex.

The molecular specificity of the IgG response against Epstein-Barr virus (EBV) was studied in 345 randomly collected sera of normal healthy individuals. The sera were tested on immunoblots containing antigens of the cell line HH514.c16 (a superinducible derivate of P3HR1), noninduced or induced for the expression of early antigens (EA) or viral capsid antigens (VCA), and from the EBV-negative cell line Ramos-Nut. This study reveals a remarkable similar antigen recognition pattern of IgG class antibodies in sera of healthy EBV carriers. The protein bands recognized predominantly have molecular weights of 18 kD, 36/38 kD, 40 kD, 72 kD, and 160 kD. The 72 kD and 36/38 kD bands were identified as EBNA1 and "Zebra," respectively, using reading frame-specific antisera. The bands at 160 kD (major capsid protein), 40 kD, and 18 kD were identified as VCA-class proteins. Of all EBV-seropositive sera tested, 98% reacted with either p18 or p40 or both. The synthesis of the antigens p18 and p40 was inhibited by phosphonoacetic acid, indicating that these were true late proteins. The detection of p18 and p40 in purified virion and capsid preparations confirms that these proteins are structural components of viral capsid antigen complex.

Purification and quantification of recombinant Epstein-Barr viral glycoproteins gp350/220 from Chinese hamster ovary cells.

Truncated Epstein-Barr virus (EBV) membrane antigen gp350/220 (EBV-MA) lacking the membrane anchor was expressed and secreted into the medium of recombinant Chinese hamster ovary cells that had been cultured in Plasmapur hollow-fibre modules using defined serum-free medium. The EBV-MA in the medium was concentrated by 70% (w/v) ammonium sulphate precipitation and subsequently purified by immunoaffinity chromatography using an anti-EBV-MA (EBV.0T6) monoclonal antibody (mAb) column. Adsorbed antigen was eluted with 3 M MgCl2 in phosphate-buffered saline, concentrated by Mono Q anion-exchange chromatography and analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, silver staining and Western blotting using EBV-positive serum and anti-EBV-MA specific mAbs. Monospecific polyclonal rabbit antibodies against the purified EBV-MA were raised and purified by protein G affinity chromatography. For the measurement of EBV-MA antigen levels a sandwich enzyme-linked immunosorbent assay using rabbit polyclonal antibodies and a horseradish peroxidase-conjugated anti-MA mAb was developed having a detection level of 10 ng/ml.

Responsiveness of the increase in C-fos mRNA levels depends on the inducer and the cell's past.

In this paper we show that in C3H10T1/2 mouse fibroblasts, the inducibility of c-fos mRNA by heat shock or serum addition is strongly dependent on the cell's past. Four hours after a heat shock, a time point where the induced c-fos mRNA has disappeared, c-fos mRNA could not be induced again by a second heat shock. Four hours after serum addition, by which c-fos was induced, a second serum addition also failed to induce c-fos mRNA again. When, however, serum was added 4 hours after heat shock or heat shock was given 4 hours after serum addition, levels of c-fos mRNA could be enhanced again. The induction by serum of c-fos mRNA levels in thermotolerant cells might be related to their increased stimulation of DNA synthesis as compared to control cells.