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Polymerase chain reaction primers specific for Ehrlichia chaffeensis were used to amplify DNA from extracts of pooled ticks. Amplification was performed on extracts from 140 pools (1,579 total ticks) consisting of three tick genera collected from five states. The characteristic 389-basepair product was observed after amplification of extracts from seven different pools of adult Amblyomma americanum (117 pools, 1,462 ticks), but not from pools of nymphs. No specific product was observed after amplification of 20 pools (105 ticks) of Dermacentor variabilis and three pools of Ixodes scapularis (12 ticks). Ehrlichia chaffeensis was present in A. americanum at a minimum frequency of > or = 0.48%, suggesting that A. americanum may be a vector of human ehrlichiosis.

Previous analysis has refined the location of the gene for Norrie disease, a severe, X-linked, recessive neurodevelopmental disorder, to a yeast artificial chromosome subfragment of 160 kilobases (kb). This fragment was used to screen cDNA libraries from human fetal and adult retina. As a result, we have identified an evolutionarily conserved cDNA, which is expressed in fetal and adult brain and encodes a predicted protein of 133 amino acids. The cDNA detects genomic sequences which span a maximum of 50 kb, and which are partly deleted in several typical Norrie disease patients. An EcoRI polymorphism with a calculated heterozygosity value of 43% was observed. The locus identified is a strong candidate for the Norrie disease gene.

It has been shown from pulsed-field gel electrophoresis (PFGE) that the monoamine oxidase genes A and B (MAOA & MAOB) and DXS7 loci are physically very close. We have therefore extended studies on their relationship through the characterisation of a 650 kb YAC isolated using L1.28 (recognising the DXS7 locus) as a probe. Restriction mapping of the YAC indicates that it contains both MAOA and MAOB genes in addition to the DXS7 locus. The map derived from the YL1.28-YAC is compatible both with the map from an independently derived YAC carrying MAOA and B genes and with the long range genomic map for the region. A series of subclones prepared from a 'phage library (lambda DASH II) of the YAC have been characterised and have been employed to determine the end point of the deletion of a Norrie disease (NDP) patient who has been shown to lack both DXS7 and MAO coding sequences. The pattern of retention of subclones in the deletion patient place the end point of the deletion within 30-130 kb of the proximal end of the YAC. By combining the data with established recombination analysis, we provide evidence that all or part of the NDP lies in the interval of approximately 250kb within the YAC.

Norrie disease is a human X-linked recessive disorder of unknown etiology characterized by congenital blindness, sensory neural deafness and mental retardation. This disease gene was previously linked to the DXS7 (L1.28) locus and the MAO genes in band Xp11.3. We report here fine physical mapping of the obligate region containing the Norrie disease gene (NDP) defined by a recombination and by the smallest submicroscopic chromosomal deletion associated with Norrie disease identified to date. Analysis, using in addition two overlapping YAC clones from this region, allowed orientation of the MAOA and MAOB genes in a 5'-3'-3'-5' configuration. A recombination event between a (GT)n polymorphism in intron 2 of the MAOB gene and the NDP locus, in a family previously reported to have a recombination between DXS7 and NDP, delineates a flanking marker telomeric to this disease gene. An anonymous DNA probe, dc12, present in one of the YACs and in a patient with a submicroscopic deletion which includes MAOA and MAOB but not L1.28, serves as a flanking marker centromeric to the disease gene. An Alu-PCR fragment from the right arm of the MAO YAC (YMAO.AluR) is not deleted in this patient and also delineates the centromeric extent of the obligate disease region. The apparent order of these loci is telomere ... DXS7-MAOA-MAOB-NDP-dc12-YMAO.AluR ... centromere. Together these data define the obligate region containing the NDP gene to a chromosomal segment less than 150 kb.

The objective of this study was to develop a continuous hydrolysis process for the enzymatic saccharification of liquefied corn starch using a membrane reactor. A residence time distribution study confirmed that the membrane reactor could be modeled as a simple continuous stirred tank reactor (CSTR). Kinetic studies indicated that the continuous reactor operated in the first-order region with respect to substrate concentration at substrate concentrations greater than 200 g/L. At a residence time of 1 h and an enzyme concentration of 1 g/L, the maximum reaction velocity (V(m)) was 3.86 g glucose/L min and the apparent Michaelis constant (K(m) (')) was 562 g/L. The K(m) (') value for the continuous reactor was 2-7 times greater than that obtained in a batch reactor.Kinetic data were fit to a model based on the Michaelis-Menten rate expression and the design equation for a CSTR. Application of the model at low reactor space times was successful. At space times of 6 min or less, the model predicted the reactor's performance reasonably well. Additional work on the detection and quantitation of reversion products formed by glucoamylase is required. Isolation, detection, and quantitation of reversion products by HPLC was difficult. Detailed analysis on the formation of these reversion products could lead to better reactor designs in the future.

Norrie disease is a rare X-linked recessive disorder characterized by blindness from infancy. The gene for Norrie disease has been localized to Xp11.3. More recently, the genes for monoamine oxidase (MAOA, MAOB) have been mapped to the same region. This study evaluates the clinical, biochemical, and neuropsychiatric data in an affected male and 2 obligate heterozygote females from a single family with a submicroscopic deletion involving Norrie disease and MAO genes. The propositus was a profoundly retarded, blind male; he also had neurologic abnormalities including myoclonus and stereotopy-habit disorder. Both obligate carrier females had a normal IQ. The propositus' mother met diagnostic criteria for "chronic hypomania and schizotypal features." The propositus' MAO activity was undetectable and the female heterozygotes had reduced levels comparable to patients receiving MAO inhibiting antidepressants. MAO substrate and metabolite abnormalities were found in the propositus' plasma and CSF. This study indicates that subtle biochemical and possibly neuropsychiatric abnormalities may be detected in some heterozygotes with the microdeletion in Xp11.3 due to loss of the gene product for the MAO genes; this deletion can also explain some of the complex phenotype of this contiguous gene syndrome in the propositus.

Recent studies are leading to a better understanding of the formation of the earth's metal core. This new information includes: better knowledge of the physics of metal segregation, improved geochemical data on the abundance of siderophile and chalcophile elements in the silicate part of the earth, and experimental data on the partitioning behavior of siderophile and chalcophile elements. Extensive melting of the earth as a result of giant impacts, accretion, or the presence of a dense blanketing atmosphere is thought to have led to the formation of the core. Collision between a planet-sized body and the earth may have also produced the moon. Near the end of accretion, core formation evidently ceased as upper mantle conditions became oxidizing. The accumulation of the oceans is a consequence of the change to oxidizing conditions.

Norrie disease is an X-linked recessive disorder characterized by congenital blindness and, in many cases, mental retardation. Some Norrie disease cases have been shown to be associated with a submicroscopic deletion in chromosomal region Xp11.3. Cerebrospinal fluid (CSF) was collected from four male patients with an X-chromosomal deletion associated with Norrie disease. CSF proteins were resolved using two-dimensional gel electrophoresis and then analyzed by computer using the Elsie V program. Our analysis revealed a protein that appears to be altered in patients with Norrie disease deletion.

Two individuals with an X-chromosomal deletion were recently found to lack the genes encoding monoamine oxidase type A (MAO-A) and MAO-B. This abnormality was associated with almost total (90%) reductions in the oxidatively deaminated urinary metabolites of the MAO-A substrate, norepinephrine, and with marked (100-fold) increases in an MAO-B substrate, phenylethylamine, confirming systemic functional consequences of the genetic enzyme deficiency. However, urinary concentrations of the deaminated metabolites of dopamine and serotonin (5-HT) were essentially normal. To investigate other deaminating systems besides MAO-A and MAO-B that might produce these metabolites of dopamine and 5-HT, we examined plasma amine oxidase (AO) activity in these two patients and two additional patients with the same X-chromosomal deletion. Normal plasma AO activity was found in all four Norrie disease-deletion patients, in four patients with classic Norrie disease without a chromosomal deletion, and in family members of patients from both groups. Marked plasma amine metabolite abnormalities and essentially absent platelet MAO-B activity were found in all four Norrie disease-deletion patients, but in none of the other subjects in the two comparison groups. These results indicate that plasma AO is encoded by gene(s) independent of those for MAO-A and MAO-B, and raise the possibility that plasma AO, and perhaps the closely related tissue AO, benzylamine oxidase, as well as other atypical AOs or MAOs encoded independently from MAO-A and MAO-B may contribute to the oxidative deamination of dopamine and 5-HT in humans.

We examined the distribution of acetylcholinesterase and nicotinamide adenine dinucleotide phosphate diaphorase enzyme activity in the human amygdala using histochemical techniques. Both methods revealed compartments of higher or lower enzyme activity, in cells or neuropil, which corresponded to the nuclear subdivisions of the amygdala as defined with classical Nissl and myelin methods. The boundaries between the histochemical compartments were usually so sharp that the identification of these nuclear subdivisions was enhanced. There was also variation of staining intensity within many of the nuclear subdivisions, such as the lateral and central nuclei, anterior amygdaloid area and the intercalated groups. This histochemical difference corresponded to more subtle differences in Nissl and myelin staining patterns, and suggests further structural subdivisions of potential functional significance. We present a revised scheme of anatomical parcellation of the human amygdala based upon serial analysis with all four techniques. Our expectation is that this will allow the delineation of a clearer homology between the cytoarchitectonic subdivisions of the human amygdala and those of experimental animals.

We report the results of two clinical trials in which patients receiving either doxorubicin (Adriamycin)/cyclophosphamide or cis-platinum cytotoxic chemotherapy were assigned at random either a single or a combination antiemetic treatment. The aim of each trial was to assess whether combination antiemetic therapy would result in improved efficacy. Sixty patients commencing doxorubicin/cyclophosphamide therapy were divided in two random groups to receive either haloperidol or haloperidol plus amitriptyline, and 80 patients commencing cis-platinum therapy were divided at random to receive either metoclopramide or metoclopramide plus promethazine. No statistically significant differences were apparent between single and combination antiemetic regimens in either of the two treatment groups. Antiemetic agents were generally well-tolerated, but minor side effects were common. The failure of our combination antiemetic regimens to intensify the proven antiemetic efficacy of single agents emphasises the need for re-evaluation of currently used antiemetic agents and their dosage schedules.

1 Pyramidal cells in rat hippocampus were used to study the molecular dimensions of a receptor for inhibitory amino acids in the central nervous system. The inhibitory potencies of gamma-aminobutyrate (GABA), beta-alanine and glycine were compared by standard microiontophoretic techniques. Subsequently, rigid cyclopentane and cyclohexane amino acid analogues were applied by iontophoresis and their relative efficacies were compared with those of the naturally occurring amino acids.2 GABA was the most effective of the small aliphatic amino acids in producing inhibition of the firing of hippocampal pyramidal neurones. beta-Alanine was less effective and glycine was the least effective. GABA-induced inhibition was antagonized by concurrent iontophoresis of picrotoxin or bicuculline, whereas strychnine did not antagonize GABA inhibition.3 The ability of the series of substituted aminocyclopentane and aminocyclohexane carboxylic acids to produce inhibition of pyramidal cells was a direct function of the separation of amino and carboxylic acid groups. In both series of the cyclic amino acids the most potent inhibition was demonstrated when the spatial separation was similar to that of the extended GABA molecule (4.74 A). Additionally, the inhibition of hippocampal pyramidal cells by (+/--cis-3-amino-cyclopentanecarboxylic acid, like that produced by GABA, could be blocked by simultaneous application of picrotoxin or bicuculline, but not by strychnine.4 The present results suggest that the physiologically active conformation of GABA is the fully extended molecule, and additionally indicate that one dimension of the postsynaptic receptor site is within the range of 4.2 to 4.8 ångströms.

Frozen red cells have been field-tested at the Mobile Fleet Hospital deployed at 29 Palms, Calif., and at Bridgeport, Calif., and at the PACOM Blood Program Office, Okinawa, Japan, to evaluate the feasibility of using these products to support the US Navy and Marines in combat areas. We studied both nonrejuvenated and rejuvenated cryopreserved red cell concentrates. Red cells stored at 4 degrees C in CPD or CPDA-1 for 3-6 days (nonrejuvenated), and red cells stored at 4 degrees C in CPD for 22-26 days or in CPDA-1 for 36-38 days before biochemical modification (rejuvenated), were frozen with 40% w/v glycerol at -80 degrees C. A multiple-bag collection system with a 600 or 800 ml primary bag was used for blood collection, and the primary bag was used for processing from preparation of the red cell concentrate through prewash dilution. The thawed red cells were washed with 1.5 liters of a sodium chloride-glucose-phosphate solution. As part of this feasibility study, frozen red cells were shipped by air in dry ice (-79 degrees C), and previously frozen washed red cells were shipped in wet ice (4 degrees C). In vitro red cell recovery values were at least 90%. After postwash storage at 4 degrees C for as long as 3 days, the red cells had 24-hour posttransfusion survivals of at least 70%, normal or slightly impaired oxygen transport function, and minimal hemolysis.

Norrie disease (ND), atrophia bulborum hereditaria, is caused by a gene defect on the proximal short arm of the X-chromosome. As shown by us and others, microdeletions spanning the DXS7 locus are not uncommon in this disorder, and there is recent evidence that, at least in some of the Norrie deletion patients, the monoamine oxidase (MAO) A and B genes are deleted as well. Molecular hybridization experiments with 19 cloned DNA fragments have enabled us to construct a preliminary long-range restriction map around DXS77, DXS7, MAO-A and MAO-B, and to localize the distal end point of an ND deletion between DXS77 and DXS7.

The long-term effectiveness of a four-week comprehensive dietary-behavioral weight control program was evaluated. Program components included a 700 kcal diet, nutritional education, medical and health education, behavior modification, and physical activity and exercise. Mean weight loss 12 months after treatment was 29.2 lb (13.2 kg) with 66 percent of patients losing 20 lb (9.1 kg) or more, 45 percent losing 30 lb (13.6 kg) or more and 29 percent losing 40 lb (18.1 kg) or more. These weight losses are higher than those generally reported for either behavioral or non-behavioral treatments. Comparison of successful versus unsuccessful patients revealed that exercise, cognitive restructuring, eating style, and social skills were most related to success.

We used a monoclonal antibody to study the immunocytochemical distribution of microtubule-associated protein 2 (MAP 2) in human fetal neocortex between the ages of 16 and 22 weeks gestation. The staining pattern was lamina-specific. Neuronal somata and dendrites in all cortical layers and in the intermediate zone were labelled. Cajal-Retzius cells of layer I, large pyramidal neurons in the inner cortical plate and neurons in the subplate were most strongly immunoreactive. Separate from the underlying cortical plate a thin sheet of small neurons in the inner marginal zone was highlighted by MAP 2 immunoreactivity. The morphologic diversity, density and regional distribution of the interstitial neurons in the subplate was emphasized by MAP 2 staining. In general, the intensity of MAP 2 immunoreactivity in cell somata and dendrites correlated with the degree of neuronal differentiation but the pattern of intracellular staining also varied as a function of laminar position, and presumably cell type.

We used in vitro autoradiography with [3H]glutamate to examine the distribution of glutamate recognition sites in 18 and 21 week gestation human fetal brains. We found a wide distribution of [3H]glutamate binding in both specimens, in a pattern distinct from that reported in adult brain using the same autoradiographic methods. In fetal brain, prominent [3H]glutamate binding was evident in hippocampal formation, caudate-putamen, globus pallidus, subthalamic nucleus, reticular nucleus of thalamus and substantia innominata.

Immunoperoxidase technique and light microscopy were used to investigate the distribution of vasoactive intestinal polypeptide (VIP) in mouse and rat brain. Both 50 micrometers unmounted cryostat and 6 micrometers deparaffinized sections were studied in coronal or sagittal plane. At least 4 different major VIP systems were found: (1) an intracerebral cortical system; (2) one innervating the central amygdala and nucleus of the stria terminalis; (3) a pathway originating in the suprachiasmatic nucleus of the hypo thalamus; and (4) another originating in the central grey of the midbrain. Specific cell body staining was seen in the limbic and neocortex, in the basal-caudal portion of the suprachiasmatic nucleus of the hypothalamus, and in the central grey of the midbrain. Heavy terminal field patterns were noted in the suprachiasmatic nucleus, central amygdaloid nucleus, bed nucleus of the stria terminalis and nucleus accumbens. Fiber density was moderate in the tuberculum olfactoriu, anterior hypothalamus including the medial preoptic area, mediobasal hypothalamus (especially dorsomedial region), periventricular thalamus, lateral lemniscal system, parabrachial nucleus, nucleus solitarius, and area postrema. Fibers could be traced dorsally from the suprachiasmatic nucleus to the dorsomedial and paraventricular nuclei of the hypothalamus and the periventricular nucleus of the thalamus. Scattered cell bodies and fibers were found in a number of other forebrain and brain stem areas with only a rare fiber seen in median eminence.

The Infra-Dry (IR) method was accurate, precise and faster than conventional (AOAC) procedures for moisture determinations in ten selected food products. The average percentage moisture by the IR method for 10 common food products and those by AOAC in parentheses are as follows: oatmeal 9.88 (9.87), cornmeal 10.86 (10.91), wheat flour 10.80 (10.80), grape nuts 4.73 (4.78), rice 12.39 (12.41), potato flakes 8.40 (8.36), bread crumbs 9.93 (9.99), noodles 9.58 (9.54), NFD milk 4.23 (4.24), and fresh milk 88.60 (88.71). The temperatures used for the IR method (135 to 155°C) were higher than those for the AOAC procedures (130°C for all products except NFD and fresh milk [100°C]); however, the times were considerably less (10 to 30 min) than for AOAC procedures (1 to 5 h). Standard deviations were generally <0.06%, except for cornmeal and noodles which were approximately 0.1%. Precisions were generally >97%. Upon removing the samples from the IR oven, results of good precision and accuracy were obtained by cooling the samples in a desiccator for 5 min or the built-in IR cooling chamber for 45 s. The recommended IR method uses a cooling chamber, is faster, and does not require a desiccator. Slightly lower precision and accuracy resulted when the entire IR oven was used. The best results were obtained using the back third of the oven. However, the slight difference in precision and accuracy is of little practical significance.

Monoamine oxidase (MAO), an important enzyme for the degradation of amine neurotransmitters, has been implicated in neuropsychiatric illness. The amino acid sequence for one form of the enzyme, MAO-A, has been deduced from human cDNA clones and verified against proteolytic peptides. The covalent binding site for the flavin adenine dinucleotide (FAD) cofactor is near the C-terminal region. The presence of features characteristic of the ADP-binding fold suggests that the N-terminal region is also involved in the binding of FAD. These cDNAs should facilitate the study of the structure, function, and intracellular targeting of MAO, as well as the analysis of its expression in normal and pathological states.

Mapping of the human MAOA gene to chromosomal region Xp21-p11 prompted our study of two affected males in a family previously reported to have Norrie disease resulting from a submicroscopic deletion in this chromosomal region. In this investigation we demonstrate in these cousins deletion of the MAOA gene, undetectable levels of MAO-A and MAO-B activities in their fibroblasts and platelets, respectively, loss of mRNA for MAO-A in fibroblasts, and substantial alterations in urinary catecholamine metabolites. The present study documents that a marked deficiency of MAO activity is compatible with life and that genes for MAO-A and MAO-B are near each other in this Xp chromosomal region. Some of the clinical features of these MAO deletion patients may help to identify X-linked MAO deficiency diseases in humans.

Multiple sclerosis (MS) is a complex disease, the symptoms of which may wax and wane on a daily basis. Conventional treatment for MS patients may involve numerous trips to a variety of clinics, physicians, and rehabilitation settings. This article describes a nurse-managed MS clinic which employs an interdisciplinary and holistic approach to treatment of MS patients with the goal of improving their quality of life and coordinating their health care. In this article, the services of the clinic are described, emphasizing the roles of the nurse practitioner (clinic manager), rehabilitation nurse specialist, and nursing director of the hospital-based home care program. A case study example of the interdisciplinary nature of this clinic's program with one MS patient is included.

The genes for MAO-A and MAO-B appear to be very close to the Norrie disease gene, on the basis of loss and/or disruption of the MAO genes and activities in atypical Norrie disease patients deleted for the DXS7 locus; linkage among the MAO genes, the Norrie disease gene, and the DXS7 locus; and mapping of all these loci to the chromosomal region Xp11. The present study provides evidence that the MAO genes are not disrupted in "classic" Norrie disease patients. Genomic DNA from these "nondeletion" Norrie disease patients did not show rearrangements at the MAOA or DXS7 loci. Normal levels of MAO-A activities, as well as normal amounts and size of the MAO-A mRNA, were observed in cultured skin fibroblasts from these patients, and MAO-B activity in their platelets was normal. Catecholamine metabolites evaluated in plasma and urine were in the control range. Thus, although some atypical Norrie disease patients lack both MAO-A and MAO-B activities, MAO does not appear to be an etiologic factor in classic Norrie disease.

Pyridoxineless mutants of Escherichia coli B stopped incorporation of nucleosides into trichloroacetic acid-insoluble material about 40 to 60 min after pyridoxine starvation was initiated, whereas incorporation of amino acids (measured the same way) slowed but did not stop for several hours. Both these incorporations and cell density were increased most effectively by the presence of either threonine or isoleucine. Arginine, glutamate, histidine, methionine, tryptophan, and tyrosine also caused significant but less dramatic increases. Inducibility of beta-galactosidase continued beyond the point where nucleic acids appeared to stop their synthesis, suggesting that messenger ribonucleic acid synthesis continued beyond ribosomal ribonucleic acid synthesis. This inducibility was also increased by isoleucine and threonine. The overall results suggest that the threonine-isoleucine biosynthetic pathway is the most sensitive to starvation for pyridoxine.