The development of writing skills in 4-year-old children with and without specific language impairment.

Research shows that many preschool children with specific language impairment (SLI) have difficulty acquiring literacy skills including phonological awareness, print concepts, and alphabet knowledge. Limited research suggests that preschool children with SLI also have difficulty with emergent writing tasks such as name writing and word writing. In typically developing children, research indicates that emergent writing skills are acquired in a developmental sequence: (1) linearity, (2) segmentation, (3) simple characters, (4) left-right orientation, (5) complex characters, (6) random letters, and (7) invented spelling. This study compared the emergent writing skills of 4-year-old children with SLI (n = 22) to their age- and gender-matched peers (n = 22). Results indicated that children with SLI demonstrate difficulty with a variety of writing tasks, including letter writing, name writing, word writing, and sentence writing when compared to their typically-developing peers. Children with SLI followed the same developmental sequence in acquiring writing skills as their typically-developing peers.

Development and certification of green tea-containing standard reference materials.

A suite of three green tea-containing Standard Reference Materials (SRMs) has been issued by the National Institute of Standards and Technology (NIST): SRM 3254 Camellia sinensis (Green Tea) Leaves, SRM 3255 Camellia sinensis (Green Tea) Extract, and SRM 3256 Green Tea-Containing Solid Oral Dosage Form. The materials are characterized for catechins, xanthine alkaloids, theanine, and toxic elements. As many as five methods were used in assigning certified and reference values to the constituents, with measurements carried out at NIST and at collaborating laboratories. The materials are intended for use in the development and validation of new analytical methods, and for use as control materials as a component in the support of claims of metrological traceability.

Common individual antigenic determinants in five of eight BALB-c IgA myeloma proteins that bind phosphoryl choline.

Eight IgA myeloma proteins derived from independently induced plasma-cytomas in genetically similar inbred BALB/c mice are functionally related by their binding of phosphoryl choline-containing antigens (Pneumococcus C polysaccharide or Lactobacillus antigen). Each protein resembles a single species of immunoglobulin in antibody. The proteins are characterized by highly sensitive myeloma-specific antisera prepared by immunizing mice of other inbred strains with the BALB/c myeloma proteins. Individual or myeloma-specific determinants located on Fab fragments were found on three of the proteins that were unique for that protein and did not react with any other IgA protein among over 70 tested. Remarkably, five of the proteins shared two common myeloma-specific determinants which were specific for this group of five proteins. These results suggest that the five functionally and genetically related proteins sharing the same myeloma-specific determinants might also be structurally similar.

Association of H-2 types with genetic control of immune reponsiveness to IgG (gamma2a) allotypes in the mouse.

Immune responsiveness to IgG allotypes in the mouse was found to be controlled by an immune response gene Ir-IgG linked to the H-2 locus. This was demonstrated by the analysis of the immune response to BALB/c IgG (gamma2a) myeloma proteins in mice of various H-2 types from five different linkage groups of immunoglobulin heavy chains. Antisera were examined for antibodies to idiotypic (Fab) and allotypic (Fc) specificities. No immune response to BALB/c IgG myeloma proteins was found in mice with the same heavy-chain immunoglobulin linkage group as BALB/c but of different H-2 types. In mice with immunoglobulin heavy chains that are different than BALB/c, a high immune response to IgG myeloma proteins was found in H-2 types b, bc, p, r, s, and v; a low response in a, d, k, and q. The Ir-IgG gene is controlled by a dominant autosomal gene.

Crossing over between genes in the immunoglobulin heavy chain linkage group of the mouse.

Immunoglobulin heavy chain genes were found in wild mice (Mus musculus) that could best be explained as recombinants of immunoglobulin genotypes. In wild mice from Kitty Hawk, N. C., two new heavy chain linkage groups, G(3,5,7,8)H(9,11)F(f)A(-) and G(3,5,8)H(9,11)F(f)A(-), were found, each of which genetically controls both the 3 and 5 distinct immunoglobulin determinants. In inbred strains the 3 and 5 determinants are found independently. The new heavy chain allotype G(3,5,7,8)H(9,11)F(f)A(-) probably arose from a homologous (intragenic) cross-over between G(3,8)H(9,11)F(f)A(-) and G(5,7,8)H(9,11)F(f)A(14) heavy chain linkage groups. It was suggested that genes controlling G(3,8)G(5,7,8), G(3,5,8), and G(3,5,7,8) are alleles. Another homozygous wild mouse (Kyushu, Japan) showed a new heavy chain allotype, (2)G(1,6,7,8)H(9,16)F(s)A(15). The 2 and G(1,6,7,8) determinants are also separated in inbred strains. The 2 determinant in inbred mice is not on the gammaF, gammaH, or gammaA heavy chain and is probably on a gammaG or gammaG-like immunoglobulin heavy chain. Papain digestion of serum from the Kyushu mouse showed two electrophoretically different Fc fragments, one carrying the G(1,6,7,8) and the other the 2 determinant. The new heavy chain allotype, (2)G(1,6,7,8)H(9,16)F(s)A(15), of the Kyushu wild mouse probably arose from a nonhomologous (unequal) cross over between (2)G(-)H(9,16)F(s)A(15) and G(1,6,7,8)H(9,11)F(f)A(12,13,14) heavy chain linkage groups. The linkage group of the Kyushu wild mouse has at least five heavy chain genes, while that of the inbred mice has four.

Clonal nature of the immune response to phosphorylcholine. I. Specificity, class, and idiotype of phosphorylcholine-binding receptors on lymphoid cells.

The relationship between receptor molecules on antigen-binding lymphocytes (ABC) and antibody produced by antibody-secreting cells was studied in inbred strains of mice using the immune response to phosphorylcholine (PC) as a model system. Splenic and lymph node lymphocytes of nonimmune mice possess rare lymphocytes which bind (125)I-labeled PC-bovine serum albumin. The frequency of PC-ABC increases after immunization and is paralleled by a rise in the frequency of PC-specific antibody-producing cells. Both of these responses are thymus independent. The receptors on these ABC display specificity for PC and are exclusively of the IgM class. In one of the strains, BALB/c, the receptors possess the same idiotype and fine degree of specificity for PC and two of its analogues, glycerophosphorylcholine and choline, that are characteristic of a PC-binding myeloma, HOPC 8. Furthermore, the idiotype and class of the receptor in these mice do not change during the course of the immune response. These data provide more direct evidence for the immunelogic relevance of receptor-bearing lymphocytes.

Multiple individual and cross-specific indiotypes on 13 levan-binding myeloma proteins of BALB/c mice.

13 leven-binding myeloma proteins (LBMP) of BALB/c origin were classified into two groups with different binding specificities; one group of 11 proteins bound beta2 leads to 1 fructosans, a second group of two proteins bound fructosans probably of beta2 leads to 6 linkage. Anti-idiotypic sera prepared to 10 of the proteins in the appropriate strains of mice identified numerous idiotypic determinants. Each protein used for immunization had its own unique individual idiotypic specificities (IdI) and in addition most of the proteins carried two-nine cross-specific or shared idiotypes (IdX) that were found only among LBMP, and not found in 106 non-LBMP. Most of the IdX determinants and only four of the IdI determinants of the beta2 leads to 1 fructosan binding group were located in the antigen-binding site. The multiplicity of antigenic differences in this functionally related group of immunoglobulins reveals an unexpected degree of heterogeneity in V-regions that appears to be unrelated to binding.

Idiotypes of inulin-binding myeloma proteins localized to variable region light and heavy chains: genetic significance.

Idiotypes of inulin-binding myeloma proteins (InuBMP) were determined primarly by variable region light chains (VL) or by variable region heavy chains (VH) but needed both chains to be expressed. Recombinant molecules were used to show that individual idiotypes (IdI) of U61, E109, T957, and A4 InuBMP and cross-specific idiotypes (IdXB) of U61 were primarily determined by VL while cross-specific idiotype (IdXA) of A4 was determined mainly by VH. The assignment of genes controlling idiotypes to VH based on allotype linkage (e.g., IdXB) is dubious until the role of the L chain in determining that idiotype is assessed. IdXB has been shown to be a VL-VH marker which presumably is controlled by two unlinked genes. However IdXB can be used as a L chain marker in combinations of strains differing in their L chain genes but having the same permissive H chain genes. Conversely IdXB can be used as a H chain marker in strains having the same permissive L chain genes but differing in their H chain genes.

H-2-linked immune response (Ir) genes. Independent loci for Ir-IgG and Ir-IgA genes.

Two H-2-linked autosomal dominant immune response (Ir) genes Ir-IgG and Ir-IgA were demonstrated to be at separate loci. Ir-IgG controls the immune response to IgG (gamma2a) myeloma proteins and Ir-IgA the immune response to IgA meyloma proteins. Both genes are associated with the H-2K region specificities of the H-2 chromosome, specifically Ir-IgG with H-2(b) and Ir-IgA with H-2(a). Different recombinants derived from H-2(a)/H-2(b) crossovers were examined for their immune responsiveness to BALB/c IgG (gamma2a) and IgA myeloma proteins. B10 (H-2(b)) parental type responded only to IgG; B10.A (H-2(a)) responded only to IgA. All the recombinants except for B10.A (4R) responded to either IgG or IgA. B10.A (4R), however, responded to both IgG and IgA. This indicated that the crossover event giving rise to B10.A (4R) occurred between the Ir-IgG and Ir-IgA loci.

Genetics of a new IgVH (T15 idiotype) marker in the mouse regulating natural antibody to phosphorylcholine.

The idiotype present on the Fab of a phosphorylcholine-binding IgA myeloma protein TEPC 15 (T15) of BALB/c origin was found in normal serum of BALB/c mice. Molecules carrying the T15 idiotype in normal serum could be adsorbed with Sepharose phosphorylcholine beads and R36A pneumococci. The T15 idiotype is absent in germ-free BALB/c but appears when the mice are conventionalized. A survey of normal sera of inbred strains for the T15 idiotype showed it to be present in BALB/c, 129, C57L, C58, and ST and absent or in low levels in CBA, C3H, C57BL/6, C57BL/Ka, C57BL/10, SJL, B10.D2, DBA/2, RIII, A, AL, AKR, NZB, and NH inbred strains of mice. The T15 idiotype is associated with some but not all strains carrying the IgC(H) allotypes found in BALB/c. Linkage of genes controlling the T15 idiotype in normal serum to the IgC(H) locus of BALB/c was demonstrated in F(2) progeny of a BALB/c and C57BL cross, Bailey's recombinant inbred strains, C x BD, C x BE, C x BG, C x BH, C x BI, C x BJ, C x BK, and CB20 congenic strains. Among these strains, only those possessing the IgC(H) locus of BALB/c including the F(2) progeny consisting of BALB/c homozygotes and BALB/c/C57BL heterozygotes and C x BG and C x BJ recombinants showed the T15 idiotype.

Ontogeny of B lymphocytes. 3. H-2 linkage of a gene controlling the rate of appearance of complement receptor lymphocytes.

The frequency of lymphocytes bearing complement receptors in the spleens of 2-wk old mice appears to be controlled by two independent genes. The presence of a "high" allele at either locus leads to intermediate or high frequency of CRL at 2 wk of age. One of the genes controlling complement receptor lymphocyte (CRL) frequency (CRL-1) is linked to the H-2 complex. Thus, in progeny of (AKR x DBA/2)F(1) x DBA/2, all mice with a low frequency of CRL at 2 wk of age are homozygous for the H-2 type of the low CRL parent (DBA/2). Furthermore, in the B10 series of congenic mice, CRL frequency at 2 wk of age is similar to the frequency in the donor of the H-2 region. Thus, C57BL/10, B10.BR, and B10-D2 mice are all of the low CRL type while B10.A mice are intermediate in CRL frequency at 2 wk. C57BR and DBA/2, the donors of the H-2 complex of the B10.BR and B10.D2, respectively, are of low CRL type while the A/WySn, the donor of the H-2 complex in the B10.A, is an intermediate CRL strain. Similarly in the A/WySn series of congenic mice, A.CA, A.SW, and A.BY are all low CRL strains while the A/WySn is intermediate. Studies of CRL frequency in mice with recombinant H-2 chromosomes (B10.A(2R), (4R), and (5R); B6/TL(+); and A/TL(-)) indicate that CRL-1 is to the right of the Ss-Slp genes and to the left of Tla.

Inability of mice with a defect in B-lymphocyte maturation to respond to phosphorycholine on immunogenic carriers.

Mice with the CBA/N defect are unresponsive to the hapten phosphorylcholine (PC) even when presented on a variety of immunogenic carriers. Since these mice have the variable region gene for PC, their inability to respond may reflect deletion or suppression of the line of B lymphocytes which is responsible for the anti-PC response.

Serum concentrations and allotypes of immunoglobulins in two lines of mice genetically selected for "high" or "low" antibody synthesis.

Random-bred Swiss mice were selectively bred for 16 generations; selection was based on their agglutinin response to sheep and pigeon erythrocytes to produce a high and a low responder line. The serum levels of individual immunoglobulins differed significantly in these two lines before immunization. The differences in the levels of immunoglobulins were much more marked after immunization with pigeon or sheep erythrocytes. Greater differences between the two lines were noted in IgM and IgG levels than in IgA. Another remarkable finding was the presence of different immunoglobulin phenotypes in the two lines. The high responders were homozygous or heterozygous for heavy-chain linkage groups found separately in the prototype BALB/c and C57BL inbred strains. The low responders were homozygous for a heavy-chain linkage group not present in bred mice in the United States, but observed as a recombinant type among wild mice probably representing a crossover between the heavy-chain linkage groups of the prototype DBA/2 and NH inbred mice.

Genetic factors controlling anti-sheep erythrocyte antibody response and immunoglobulin synthesis in backcross and F2 progeny of mice genetically selected for "high" or "low" antibody synthesis.

Agglutinin responses to sheep erythrocytes and immunoglobulin heavy chain phenotypes determined in F(1), F(2), and backcross progeny of mice genetically selected for high and low antibody synthesis indicated that an immune response gene for sheep erythrocytes is linked to the immunoglobulin heavy chain allotype. Mice homozygous for the phenotype of the high line had significantly higher titers than mice homozygous for the phenotype of the low line. An association was also observed in some progeny of the backcross of the F(1) generation with the low line. However, the control of the immune response was clearly multigenic since heterozygous mice of the same phenotype (2/3, 5) resulting from the two backcrosses (high and low) had very different immune responses. Immunoglobulin levels in the same progeny showed no linkage to the immunoglobulin allotype but a rather simple pattern of inheritance.

Regulation of the anti-inulin antibody response by a nonallotype-linked gene.

The antibody response to the inulin [(In), beta-(2 leads to 1) fructosan] determinant of bacterial levan [(BL), a beta-(2 leads to 6) polyfructosan that contains beta-(2 leads to 1) branch points] requires the presence of the a haplotype of the Igh gene complex. BALB/c (Igh a) mice immunized with BL produce IgG anti-In antibodies of a single spectrotype by isoelectric focusing analysis. C57BL/6 mice, which possess the b haplotype of the Igh gene complex and which fail to produce anti-In antibodies, nevertheless possess a gene, spectrotype regulation gene 1 (Sr-1), that regulates the isoelectric focusing (IEF) pattern of anti-In antibodies in mice of the a haplotype. Thus, the IEF patterns of anti-In antibodies of (BALB/c x C57BL/6)F1 mice and of B.C8 mice (C57BL/Ka . Igh-Ca) are considerably more complex than those of BALB/c. Backcross analysis indicates that Sr-1 is not linked to the Igh complex, the major histocompatibility complex, or to the genes that code for coat color. Studies of the heterogeneity of anti-In antibodies in recombinant inbred lines and their progeny from matings to BALB/c and C.B20 (BALB/c . Igh-Cb) suggest the existence of other regulatory genes.

Introduction of H-2Dd determinants into the H-2Ld antigen by site-directed mutagenesis.

We used site-directed mutagenesis to localize serologically defined (s) and CTL (c)-defined alloantigenic determinants to discrete amino acid sequences of a murine MHC class I antigen. Based on the prediction that amino acid position 63-73 of the H-2Dd antigen forms s-allodeterminants, the H-2Ld gene was mutated in a sequential fashion to replace codons for amino acid positions 63, 65, 66, 70, and 73 with those of the H-2Dd amino acids. Epitopes of the mutant antigens expressed in L-cells were examined by the binding of a series of mAbs specific for the H-2Dd antigen. The mutant antigen M66 had substitutions at residues 63, 65, and 66, and resulted in the acquisition of a number of H-2Dd-specific s-epitopes. Mutant M70 had an additional substitution at residue 70, which led to the gain of multiple additional H-2Dd s-epitopes. Together, more than half of all the relevant H-2Dd s-epitopes were mapped into amino acid position 63-70 of the H-2Dd molecule, which was expressed in the mutant H-2Ld gene. The final mutation at residue 73 (M73) caused no new epitope gains, rather, a few Dd s-epitopes acquired by the preceding mutations were lost. All of the H-2Ld-specific s-determinants were retained in the mutant molecules, as were H-2Dd s-determinants specific for the alpha-2 or alpha-3 domains. Changes of these residues affected c-determinants defined by CTL. Anti-H-2Dd CTL cultures and an anti-H-2Dd CTL clone recognized the mutant H-2Ld molecules, M66 and M70. Some CTL clones generated against the Q10d molecule, which has an identical sequence to H-2Dd between residues 61 and 73, failed to recognize native H-2Dd or Ld but did crossreact with mutant Ld. While bulk-cultured anti-H-2Ld CTL cultures reacted strongly against M73, bulk-cultured H-2Ld restricted anti-vesicular stomatitis virus CTL did not. Finally, at the clonal level two of three anti-H-2Ld CTL clones lost reactivity with some or all of these mutant molecules. From these results we conclude that a stretch of amino acids from position 63 to 70 of the alpha-1 domain controls major s- and c-antigenic sites on the H-2Dd antigen and c-sites on H-2Ld antigen.

Oestrogen-mediated phosphorylation and stabilization of BRCA2 protein in breast.

Disease-associated BRCA2 mutations typically result in protein truncations that delete the phosphorylation-regulated S3291 BRCA2 domain that interacts with Rad51. BRCA2 hereditary breast cancers are usually ER(+), differing from BRCA1 hereditary cancers, which are usually ER(-). We studied BRCA2 protein expression and S3291 phosphorylation in normal breast tissues and in sporadic breast cancers and observed that BRCA2 is expressed and phosphorylated in normal breast and 10 ER(+) breast cancers but not in 10 ER(-) breast cancers. In order to study this correlation between ER and BRCA2 expression, we studied ER(+) breast cancer cell lines. We found that a rapid increase in BRCA2 S3291 phosphorylation occurs following 17-beta-oestradiol (E2) treatment. This increase seen in BRCA2 total and phospho-S3291 protein levels was found to be unaffected with cycloheximide pre-treatment, but decreased following tamoxifen, ICI 182,780 or roscovitine treatment. This suggests a requirement for ER and cdk (cyclin-dependent kinase) in mediating the increased protein levels. MCF7 cell cycle distribution analysis following E2, in both the presence and absence of roscovitine (a cdk inhibitor), did not demonstrate any changes during an 8 h period, which further supports our hypothesis that mitogenic effects of E2 are not predominant at early time points. Studies with MG132 proteasome inhibitor and siRNA to skp2 support a model in which skp2-mediated proteasomal degradation of BRCA2 rapidly degrades BRCA2 protein in the absence of hormone treatment, which likely inhibits this pathway. E2 was shown to improve survival of MCF7 cells upon radiation treatment and roscovitine partially reversed this effect. We have demonstrated that BRCA2 protein is specifically expressed in ER(+) breast cancers and are investigating a pathway that may show a link between E2 action and BRCA2 protein function in breast cancer.

Polymorphism of heavy-chain genes in immunoglobulins of wild mice.

The serums of 123 wild mice from six different geographic locations in the United States contain five of the six known heavy-chain antigenic determinants that have been identified in immunoglobulin of inbred laboratory strains of mice. On the basis of the distribution of determinants in inbred strains,44 of the mice were judged to be heterozygotes of various combinations and two had combinations of determinants that were unusual and could only have occurred in laboratory inbred mice by recombination.

2-chain immunoglobulin A molecules: abnormal or normal intermediates in synthesis.

Immunoglobulin A (gammaA) myeloma proteins secreted by plasma-cell tumors of mice are of two types, a common four-chain molecule and a rare two-chain (3.9S) molecule. The close similarity between two-chain gammaA molecules and four-chain gammaA molecules and their polymers is demonstrated in tryptic peptide maps of isolated polypeptide chains and by precipitin reactions with rabbit antiserums to gammaA immzunoglobulins. However, a difference between these two types is distinguishable with homologous antiserums. Homologous antiserums to two-chain gammaA immunoglobulins are specific and do not cross-react with four-chain gammaA immutnoglobulins.

The Contributions of Phonological Awareness, Alphabet Knowledge, and Letter Writing to Name Writing in Children With Specific Language Impairment and Typically Developing Children.

Purpose: Name writing is one aspect of emergent writing that has been used to understand emergent literacy development. Name-writing skills and the relationship of name writing to other emergent literacy skills have not been studied extensively in children with specific language impairment (SLI). Children with SLI consistently demonstrate delays in phonological awareness (PA), alphabet knowledge (AK), print awareness, and emergent writing. The purpose of this study was to examine the contributions of PA, AK, and letter writing to name writing in children with SLI and typically developing (TD) children. Method: Participants were 65 children (22 SLI, 43 TD) with an average age of 53 months. Participants completed the Assessment of Literacy and Language (Lombardino, Lieberman, & Brown, 2005), a letter-writing task, and a name-writing task. Results: Data were analyzed using correlation and mediation modeling. Mediation modeling, a more sophisticated analysis, revealed that PA, AK, and letter writing, in serial, were mediating variables for language status on name writing. Conclusion: Phonemic awareness, AK, and letter writing help to explain the relationship between language status and name writing. These skills should be integrated during treatment, using a horizontal approach with developmentally appropriate activities, particularly for children with SLI.