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Objective To determine the predictive value of serum concentrations of total protein (sTP), albumin (sAlb), and globulin (sGlob) measured by automated calorimetric assays to estimate serum immunoglobulin G (sIgG) concentrations in neonatal foals and identify failure of transfer of passive immunity when compared to turbidoimmunometric assay determinations of sIgG. Design Retrospective and prospective analysis of laboratory data. Setting University tertiary care facility. Animals Group 1 (retrospective): foals (n = 45) ≤7 days of age in which sIgG, sGlob, sAlb, and sTP concentrations were measured on an automated chemistry analyzer. Group 2 (prospective): foals (n = 31) ≤7 days of age with same laboratory data collected used to validate equations generated from group 1 foals. Interventions Spearman rank correlations between measured sIgG and serum protein concentrations were performed. When significant correlation was found, sIgG was estimated using an sGlob simple linear regression and estimated using a sGlob, sTP, and sAlb multiple linear regression. Comparisons between estimated and measured sIgG was performed using Kruskal-Wallis testing. Sensitivity, specificity, positive predictive value, and negative predictive value were calculated to identify foals with sIgG < 8.0 g/L [

Objectives To determine whether direct intra-abdominal pressures (IAP) and calculated direct abdominal perfusion pressures (APP) are location dependent within the abdomen of standing horses. We hypothesize that IAP will be increased and calculated APP will be decreased at a ventral abdominal location (V) when compared to values obtained from the left (LFl) or right flank (RFl). Design Prospective experimental design. Setting University-based equine research facility. Animals Seven healthy adult horses, 4 geldings and 3 mares. Interventions Measurements of direct IAP obtained from the RFl, LFl, and V locations via abdominal cannulation and direct arterial blood pressures obtained via catheterization of the transverse facial artery were obtained in fasted, standing, unsedated horses. APP was calculated for each location by the subtraction of IAP from the mean arterial pressure. Differences between sites of measurement for IAP, APP, and their gradients were calculated and compared by ANOVA and t-tests. Measurements and Main Results Mean flank IAP measurements were subatmospheric and negative compared to ventral IAP values (LFl = –3 mm Hg, RFl = –5 mm Hg, V = 25 mm Hg; P < 0.001 between each flank and the ventral location). Ventrum APP was lower than flank APP (V = 82 mm Hg; LFl = 106 mm Hg; RFl = 108 mm Hg; P = 0.029 between each flank and the ventral location). Gradient calculations between sites showed the IAP increased and APP decreased from dorsal to ventral (P < 0.05) and from right to left (P = 0.004) within the abdomen. Conclusions IAP and calculated APP are location dependent. These data provide new information regarding abdominal pressure profiles in standing healthy adult horses.

The inherent neurotoxic potential ofthe endogenous excitatory amino acid glutamate, may be causally related to the pathogenesis ofAD neurodegeneration disorders. Neuronal excitotoxicity is conceivably mediated by the N-methyl-D-aspartate-(NMDA)-Ca2+- ionotropic receptor. NMDA receptors exist as multimeric complexes comprising proteins from two families – NR1 and NR2(A-D). The polyamines, spermine and spermidine bind to, and modulate NMDA receptor efficacy via interaction with exon 5, an alternatively-spliced, 21 amino acid, N-terminal cassette. ADassociated cognitive impairment may therefore occur via subunitspecific NMDA receptor dysfunction effecting regional selectivity ofneuronal degradation. Total RNA was prepared from pathologically spared and susceptible regions from AD cases and matched controls. Quantitation was performed using standard curve methodology in which a known amount ofa synthetic ribonucleic acid competitor deletion construct was co-amplified against total RNA. Expression profile analysis oftwo NR1 mRNA subsets has revealed significant differences in NR11XX mRNA levels in cingulate gyrus, P.

It is generally assumed that rodent brains can be used as representative models of neurochemical function in other species, such as humans. We have compared the distributions of the predominant glial glutamate transporters in rodents, rabbits, cats, pigs, monkeys, and humans. We identify similarities but also significant differences between species. GLT-1v, which is abundantly expressed by rodent astrocytes, is expressed only in a rare subset of astrocytes of cats and humans, and appears to be absent from brains of rabbits and monkeys. Conversely, in the pig brain GLT-1v is expressed only by oligodendrocytes. GLAST and GLT-1 expression differed significantly between species; while rodents and rabbits exhibited uniform expression patterns in cortex, higher species, including cats, pigs, monkeys, and humans, exhibited heterogeneities in cortical and hippocampal expression. Patches devoid of labeling intermingling with patches of strong labeling were evident in areas such as temporal cortex and frontal cortex. In addition, we noted that in human motor cortex, there were inconsistencies in labeling for the C-terminal of GLT-1 and common domains of GLT-1, suggesting that the C-terminal region may be missing or that an unidentified splicing is present in many human astrocytes. Collectively our data suggest that assumptions as to the roles of glutamate transporters in any species may need to be tested empirically. © 2004 Wiley-Liss, Inc.

Previous work had shown that the ratio of NMDA receptor NR1 subunit mRNA transcripts containing an N-terminal splice cassette to those that do not is markedly lower in regions of the Alzheimer's disease (AD) brain that are susceptible to pathological damage, compared with spared regions in the same cases or homotropic regions in controls. To elucidate the origins of this difference in proportionate expression, we measured the absolute levels of each of the eight NR1 transcripts by quantitative internally standardized RT-PCR assay. Expression of transcripts with the cassette was strongly attenuated in susceptible regions of Alzheimer's brain, whereas expression of non-cassette transcripts differed little from that in controls. The expression of other NR1 splice variants was not associated with pathology relevant to disease status, although some combinations of splice cassettes were well maintained in AD cases. The population profile of NR1 transcripts in occipital cortex differed from the profiles in other brain regions studied. Western analysis confirmed that the expression of protein isoforms containing the N-terminal peptide was very low in susceptible areas of the Alzheimer's brain. Cells that express NR1 subunits with the N-terminal cassette may be selectively vulnerable to toxicity in AD.

The use of human brain tissue obtained at autopsy for neurochemical, pharmacological and physiological analyses is reviewed. RNA and protein samples have been found suitable for expression profiling by techniques that include RT-PCR, cDNA microarrays, western blotting, immunohistochemistry and proteomics. The rapid development of molecular biological techniques has increased the impetus for this work to be applied to studies of brain disease. It has been shown that most nucleic acids and proteins are reasonably stable post-mortem. However, their abundance and integrity can exhibit marked intra- and intercase variability, making comparisons between case-groups difficult. Variability can reveal important functional and biochemical information. The correct interpretation of neurochemical data must take into account such factors as age, gender, ethnicity, medicative history, immediate ante-mortem status, agonal state and post-mortem and post-autopsy intervals. Here we consider issues associated with the sampling of DNA, RNA and proteins using human autopsy brain tissue in relation to various ante- and post-mortem factors. We conclude that valid and practical measures of a variety of parameters may be made in human brain tissue, provided that specific factors are controlled.

The displacement of Na+-dependent D-[H-3]aspartate binding by unlabeled D-aspartate or the inhibitors DL-threo-beta-hydroxyaspartate, L-cysteate, L-glutamate, dihydrokainate, DL-alpha-aminoadipate, alpha-methyl-DL-gIutamate, and 1-aminocyclobutane-cis-1,3-dicarboxylate was used to characterize the high-affinity glutamate/aspartate uptake site in human cerebral cortex. Synaptosomal membranes were prepared from tissue obtained at autopsy from nondemented control, Alzheimer's disease (AD), and diffuse Lewy body disease (DLBD) cases, Areas that are damaged in AD (midtemporal, frontal, caudal cingulate, and hippocampal cortices) were compared with those that are spared (occipital and motor cortices), Profiles of the affinities (K-a values) of the ligands showed that areas spared from damage in AD cases differed significantly from equivalent areas in control (p < 0.001) and DLBD (p < 0.001) cases and also from areas susceptible to damage in the same AD cases (p < 0.001). Areas susceptible to damage in AD showed comparable profiles across the three case groups (p = 0.980). The glutamate/aspartate uptake site may be regionally variant in AD cases, and this may underlie local excitotoxicity, D-[H-3]Aspartate binding site density was significantly lower in both dementia groups (control vs, AD, p ( 0.001; control vs. DLBD, p = 0.009; but AD vs, DLBD, p = 0.528); within-group differences were not significant (control, p = 0.874; AD, p = 0.285; DLBD, p = 0.741).

We analyzed the expression profile of two NMDAR1 mRNA isoform subsets, NR1(0XX) and NR1(1XX), in discrete regions of human cerebral cortex. The subsets are characterized by the absence or presence of a 21-amino acid N-terminal cassette. Reverse transcription polymerase chain reaction for NR1 isoforms was performed on total RNA preparations from spared and susceptible regions from 10 pathologically confirmed Alzheimer's disease (AD) cases and 10 matched controls. Primers spanning the splice insert yielded two bands, 342 bp (NR1(0XX)) and 405 bp (NR1(1XX)), on agarose gel electrophoresis. The bands were visualized with ethidium and quantified by densitometry. NR1(1XX) transcript expression was calculated as a proportion of the NR1(1XX) + NR1(0XX) total. Values were significantly lower in AD cases than in controls in mid-cingulate cortex, p < 0.01, superior temporal cortex, p < 0.01 and hippocampus, p approximately 0.05. Cortical proportionate NR1(1XX) transcript expression was invariant over the range of ages and areas of controls tested, at approximately 50%. This was also true for AD motor and occipital cortex. Proportionate NR1(1XX) expression in AD cingulate and temporal cortex was lower at younger ages and increased with age: this regression was significantly different from that in the homotropic areas of controls. Variations in NR1 N-terminal cassette expression may underlie the local vulnerability to excitotoxic damage of some areas in the AD brain. Alternatively, changes in NR1 mRNA expression may arise as a consequence of the AD disease process.