Genomic organisation and sequence of the extracellular domain exons of the bovine Fc gamma RI receptor, and evidence for restricted binding of ruminant IgG to U937 cells.

Southern blotting of bovine genomic DNA and hybridization with a human Fc gamma RI cDNA probe, p135, has identified a single copy of the bovine Fc gamma RI gene. A bovine genomic lymphocyte library in lambda EMBL3 was screened with probe p135. A positive lambda clone, 15.5.4, containing the three extracellular domain exons of Fc gamma RI, has been cloned, mapped and sequenced. Each extracellular domain is encoded within a single exon. All three domains are assigned to the C-2 set of the Ig superfamily with 58% identity between amino acid residues of bovine, human and mouse Fc gamma RI. Pairs of cysteine residues are conserved in each domain as potential sites for intra-chain disulphide bonding. Human monocytoid U937 cells were used as a model to test binding homology within the Fc gamma RI family. The binding of IgG isotypes to IFN-gamma stimulated U937 cells was determined by FACScan flow cytometry. U937 cell Fc gamma RI receptor does not bind bovine or ovine IgG isotypes. On the basis of these studies and by comparison of the Fc determinant region sequences of IgG, the introduction of species specificity in Fc gamma RI/IgG interaction by evolutionary drift is proposed.

Structure of bovine immunoglobulin constant region heavy chain gamma 1 and gamma 2 genes.

Two bovine immunoglobulin constant region gamma heavy chain germline gene sequences are described. A gamma 1 gene was cloned from a lambda 2001 calf liver library screened with a human gamma 4 (pBRH4.1) probe and is contained in a 5.8 kb BamH1 hybridizing fragment. The gamma 2 gene was from an EMBL4 lambda library and is in a 6.6 kb BamH1 fragment. Each of these genes is arranged in four exons corresponding to the three CH domains and the hinge of gamma heavy chain genes; normal RNA splice and polyadenylation sites are present. The translated C-terminal peptide sequences of the genes match exactly the equivalent peptide sequences of bovine IgG1 and IgG2 heavy chains, identifying them as gamma 1 and gamma 2. The derived protein sequences reveal 96, 80 and 83% identity of amino acid residues between their CH1, CH2 and CH3 domains. Two adjacent cysteine residues encoded in the CH1 exons suggest that, as in rabbit gamma, an extra intra-chain disulphide bond occurs in the bovine gamma heavy chains. Significant DNA rearrangement in the hinge-CH2 region is evident in the bovine gamma 2 gene, with resultant deletion and substitution of amino acid residues in the lower hinge and N-terminal portion of the CH2 domain. The Fab-Fc interface of bovine IgG2 is predicted to be sterically blocked, relative to IgG1, which has implications for effector differences between the bovine gamma subclasses.

Sequence of ovine Ig gamma 2 constant region heavy chain cDNA and molecular modelling of ruminant IgG isotypes.

Ovine mesenteric lymph node mRNA was used for PCR amplification of DNA coding for immunoglobulin gamma 1 and gamma 2 heavy chain constant regions. Primers complementary to regions of CH1 conserved between ruminants were used for upstream priming, with downstream priming on the poly-A segment. PCR products of the appropriate length were cloned and gamma positive clones selected with a CH1 conserved-region probe. Of these, gamma 1 clones were positively selected and gamma 2 clones negatively selected with a gamma 1 hinge-specific probe. Ovine gamma 2 cDNA has 93% identity of nucleotides with ovine gamma 1. Both ovine gamma 1 and gamma 2 CH1 domains encoded two consecutive cysteine residues (Cys-127, -128, Kabat numbering), an arrangement which is deduced to form a pair of disulphide bridges, one to the L chain and one as an intra-chain bridge to the uppermost Cys of the hinge, as in rabbit and goat IgG. The majority of the differences between the isotypes occur in the hinge region and an evolutionary pattern for ruminant IgG hinges can now be identified. IgG1 isotypes are typical, with hinges containing the C-terminal Cys-Pro motif, but deletion and replacement of nucleotides (in the ancestral gene) of ruminant gamma 2 has shortened the IgG2 hinge, removing the Cys-Pro motif and the consensus high affinity Fc gamma RI receptor motif at the start of CH2. An N-terminal glycosylation site and the peptide motif for complement C1q binding are present in CH2 of both isotypes. The hinge regions of gamma 1 and gamma 2 and predicted structures for ovine IgG1 and IgG2 have been modelled. Close apposition of Fab and Fc in IgG2 produces steric hindrance at the normally accessible Fab/hinge/Fc interface; the structural differences between the ruminant isotypes form a basis for understanding some of the differences in their effector properties.

A method for the selection of pig lymphocytes bearing surface immunoglobulin.

Coupling of IgG anti-immunoglobulin to polystyrene bottles with carbodiimide has been used to bind selectively pig lymphocytes previously treated with anti-immunoglobulin or anti-Ig class-specific fluorescent conjugates. Subsequent recovery of the bound cells produced populations with approximately 80% of cells stained for surface Ig and 90% viability.

The stuff that dreams aren't made of: why wake-state and dream-state sensory experiences differ.

It is adaptive for individuals to be continuously alert and responsive to external stimuli (such as the sound and odor of an approaching predator or the cry of an infant), even during sleep. Natural selection thus has disfavored the occurrence during sleep of hallucinations that compromise external vigilance. In the great majority of mammalian species, including Homo sapiens, closed eyes and immobility are basic aspects of sleep. Therefore, (a) visual and movement sensory modalities (except kinesthesis) do not provide the sleeper with accurate information about the external environment or the sleeper's relationship to that environment; (b) the sleeper's forebrain "vigilance mechanism" does not monitor these modalities; hence (c) visual and movement hallucinations--similar or identical to percepts--can occur during sleep without compromising vigilance. In contrast, the other sensory modalities do provide the sleeper with a continuous flow of information about the external environment or the sleeper's relationship to that environment, and these modalities are monitored by the vigilance mechanism. Hallucinations of kinesthesis, pain, touch, warmth, cold, odor, and sound thus would compromise vigilance, and their occurrence during sleep has been disfavored by natural selection. This vigilance hypothesis generates novel predictions about dream phenomenology and REM-state neurophysiology and has implications for the general study of imagery.

Time perception: does it distinguish ADHD and RD children in a clinical sample?

This study used a double-dissociation design to evaluate whether children with ADHD demonstrated specific deficits relative to children with Reading Disorders. Recent theory suggests that ADHD children have deficits in time perception and working memory, whereas RD children have deficits in phonological decoding. The performance of 113 clinic-referred children aged 6-11 was examined using measures of working memory, phonological processing, and time perception. Respondents completed two time production tasks in which they were to judge when 30-s had elapsed, and another in which they were asked to estimate the duration of the Conners' CPT (CCPT). Time Perception and phonological processing variables were submitted to a 2 x 2 ANCOVA (ADHD vs. RD), covarying for age, SES, IQ, and working memory. Children with ADHD were more likely to overestimate the time taken for the CCPT than children without ADHD, but no group differences were found on the 30-s estimation tasks. Children with RD did not display deficits in time estimation, but showed deficits in auditory phonological processing. The lack of interaction effects supported an "etiological subtype" over the "phenocopy" model of ADHD and RD. No group differences were detected using the CCPT. Although our previous studies did not find an order effect for the Conners' CPT in a 1-hr battery, a fatigue effect was evident with a 1.5-hr battery. The implications for Barkley's behavioral inhibition theories (R. Barkley, 1997) are discussed.

Does the Conners' Continuous Performance Test aid in ADHD diagnosis?

The performance of clinic-referred children aged 6-11 (N = 100) was examined using the Conners' Continuous Performance Test (CPT) and measures of auditory attention (Auditory Continuous Performance Test; ACPT), phonological awareness, visual processing speed, and visual-motor competence. The Conners' CPT overall index was unrelated to measures of visual processing speed or visual-motor competence. Although the Conners' CPT converged with the ACPT, the latter demonstrated age and order effects. Significant variance in Conners' CPT parameters was predicted by phonological awareness measures, suggesting that Reading Disordered (RD) children could be "false positives" on the Conners' CPT. The Conners' CPT overall index, phonological awareness, and visual-motor measures were submitted to a 2 x 2 MANCOVA (ADHD vs. RD, covarying for age and socioeconomic status): a main effect for RD status was found. Children with ADHD did not have higher Conners' CPT scores than did clinical controls; however, children with Reading Disorders did. Phonological measures distinguished RD children from ADHD children and other clinical controls. ADHD children who failed the Conners' CPT were rated by teachers as more hyperactive. Despite the strengths of the Conners' CPT, its utility for differential diagnosis of ADHD is questioned.

The bidirectional relationship between achievement and externalizing behavior problems of students with learning disabilities.

The current study examined the bidirectional relationship between academic achievement and externalizing behavior problems of adolescents with learning disabilities. Forty-three students attending a residential school were assessed for externalizing behavior problems via parent and teacher reports on the Child Behavior Checklist and the Children's Attention and Adjustment Survey. The Wide Range Achievement Test-Revised was used to estimate academic achievement in reading, spelling, and arithmetic. Achievement measures did not predict externalizing behavior problems 1 to 2 years later, although verbal IQ predicted parent reports. Teacher reports of externalizing behavior problems predicted reading and spelling achievement scores 1 year later, and parent reports of externalizing behavior problems predicted changes in reading achievement. There was evidence that attentional problems were the component of externalizing behavior accounting for these relations, consistent with the explanation that inattentive students have difficulty achieving in an intensive learning environment.

Pulpal obliteration related to long-term glucocorticosteroid medication.

Glucocorticosteroids (GCS) are used in the treatment of chronic autoimmune diseases and many long-term illnesses. While these drugs may be considered essential for the patient's health, adverse reactions do occur. The incidence of adverse reactions depends upon the dose level and the duration of treatment. Three case reports of patients on long-term GCS therapy are presented. Pulpal obliteration was assessed by radiographic records. In all instances, pulpal obliteration to some degree is described; however, the rate and extent of pulpal obliteration in the mature dentition seem greater than those occurring in the immature dentition. In the cases reported, the greatest dentin deposition appeared to occur on the roof of the pulp chamber, indicating that the pattern of dentin deposition by pulp tissue in GCS-treated individuals is different from that of the normal aging process.

Ultrastructural characterization of the mesenchyme of the facial processes during development of the rat intermaxillary segment.

This study quantifies ultrastructural changes within the mesenchyme of the maxillary and medial/lateral nasal processes before and after formation of the intermaxillary segment. At both day 11 and day 13 of development, 4 fetal rat heads were fixed with 3% glutaraldehyde and processed for electron microscopy. Differences between cells within the facial processes were discerned at day 11 of gestation. Significantly more rough endoplasmic reticulum was present in the cells of the maxillary processes than in the lateral nasal processes (p < .05), and a greater number of cell projections was found in the maxillary processes than in the medial nasal processes (p < .05). At day 13, no significant differences were found in the facial processes. Of particular note, a consistent increase in the number of cell projections within all the facial processes was found on day 13 when compared with day 11 (p < .05). This change may be related to intercellular signaling between the mesenchymal cells at the time of onset of major histogenic changes.

Distribution of NADPH-diaphorase activity in rat paravertebral, prevertebral and pelvic sympathetic ganglia.

Paravertebral (superior cervical and stellate), prevertebral (coeliac-superior mesenteric, inferior mesenteric) and pelvic (hypogastric) sympathetic ganglia of the rat were investigated by enzyme histochemistry to ascertain the distribution of nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-diaphorase) activity. In the paravertebral ganglia the majority of the sympathetic neuronal perikarya contained lightly and homogeneously distributed formazan reaction product but there was a range of staining intensities amongst the neuron population. In contrast, in the prevertebral ganglia, intense NADPH-diaphorase staining was present in certain neurons. Firstly, a population of neurons of the coeliac-superior mesenteric ganglion complex were surrounded by densely NADPH-diaphorase-positive 'baskets' of fibres and other stained fibres were seen in interstitial nerve bundles and in nerve trunks connected to the ganglion complex. Secondly, in both the inferior mesenteric ganglion and hypogastric ganglion there were many very intensely NADPH-diaphorase positive neurons. Stained dendritic and axonal processes emerged from these cell bodies. In both ganglia this population of neurons was smaller in size than the lightly stained ganglionic neurons and commonly had only one long (presumably axonal) process. The similarity of these highly NADPH-diaphorase-positive neurons with previously described postganglionic parasympathetic neurons in the hypogastric ganglion is discussed.

Immunoglobulin-bearing lymphocytes: their demonstration in adult sheep and ontogeny in the sheep fetus.

A method for the preparation of lymphocytes from sheep blood is described. Lymphocytes from adult and fetal sheep have been examined by immunofluorescence for surface immunoglobulin (sIg) determinants. Young adults (1.5 years) had a mean of 23.2% sIg cells, older adults (8.75 years) a mean of 9.7% sIg cells. Pregnancy did not influence these values. The earliest lymphocytes with sIg in fetal lambs were demonstrable at 52 days (96 mm crown-rump length) and countable by 56 days (110 mm CRL) at 0.3% sIg. The percentage rose rapidly to 15.1% between 78 and 87 days, followed by a fall to 2.1% around 117 days with a second increase thereafter. The significance of this spontaneous appearance of fetal sIg cells is discussed.

A simple aid in the evaluation of glass knives.

This report describes a compact, portable, self-contained light-box for the qualitative inspection of glass knives under a microscope. It obviates the need for elaborate illumination and viewing facilities in the microtomy laboratory when the ultramicrotome is not itself equipped with knife-inspection devices.

The binding of LPS to the lymphocyte surface.

Bacterial lipopolysaccharide (LPS) from Escherichia coli labelled with tritium has been used to follow the binding of LPS to lymphocytes. Binding to cells rose to a maximum 2-7 min after addition of [3H]-LPS, followed by loss of [3H]-LPS from cells, reducing to about 10% of the peak level at 20-30 min. Peripheral blood lymphocytes, mesenteric lymph node and thymus cells of the pig and CBA, C3H/He and C3H/HeJ mouse spleen cells all bound [3H]-LPS transiently at similar levels. It is concluded that this type of LPS binding cannot be solely responsible for the preferential stimulation of B cells by LPS.