Clostridium perfringens enterotoxin is a superantigen reactive with human T cell receptors V beta 6.9 and V beta 22.

Candidate superantigens were screened for their ability to induce lysis of human histocompatibility leukocyte antigen class II-positive targets by human CD8+ influenza-specific cytotoxic T cell (CTL) lines. Clostridium perfringens enterotoxin (CPET) induced major histocompatibility complex unrestricted killing by some but not all CTL lines. Using "anchored" polymerase chain reactions, CPET was shown to selectively stimulate peripheral blood lymphocytes bearing T cell receptor V beta 6.9 and V beta 22 in five healthy donors. V beta 24, V beta 21, V beta 18, V beta 5, and V beta 6.1-5 appeared to be weakly stimulated. Antigen processing was not required for CPET to induce proliferation. Like the staphylococcal enterotoxins, CPET is a major cause of food poisoning. These data suggest that superantigenic and enterotoxigenic properties may be closely linked.


Inhibition of proliferation of normal and transformed neural cells by blood group-related oligosaccharides.

A synthetic tetrasaccharide structurally related to blood groups and selectin ligands inhibited division of astrocytes, gliomas, and neuroblastomas at micromolar concentrations. The compound was cytostatic for primary astrocytes in culture, but cytotoxic for fast proliferating cell lines.


Cytosolic protein concentration is the primary volume signal for swelling-induced [K-Cl] cotransport in dog red cells.

Chloride-dependent K transport ([K-Cl] cotransport) in dog red cells is activated by cell swelling. Whether the volume signal is generated by a change in cell configuration or by the dilution of some cytosolic constituent is not known. To differentiate between these two alternatives we prepared resealed ghosts that, compared with intact red cells, had the same surface area and similar hemoglobin concentration, but a greatly diminished volume. Swelling-induced [K-Cl] cotransport was activated in the ghosts at a volume (20 fl) well below the activation volume for intact cells (70 fl), but at a similar hemoglobin concentration (30-35 g dry solids per 100 g wet weight). Ghosts made to contain 40% albumin and 60% hemoglobin showed activation of [K-Cl] cotransport at a concentration of cell solids similar to intact cells or ghosts containing only hemoglobin. [K-Cl] cotransport in the resealed ghosts became quiescent at a dry solid concentration close to that at which shrinkage-induced Na/H exchange became activated. These results support the notion that the primary volume sensor in dog red cells is cytosolic protein concentration. We speculate that macromolecular crowding is the mechanism by which cells initiate responses to volume perturbation.


Effects of low myoplasmic Mg2+ on calcium binding by parvalbumin and calcium uptake by the sarcoplasmic reticulum in frog skeletal muscle.

The effects of low intracellular free Mg2+ on the myoplasmic calcium removal properties of skeletal muscle were studied in voltage-clamped frog skeletal muscle fibers by analyzing the changes in intracellular calcium and magnesium due to membrane depolarization under various conditions of internal free [Mg2+]. Batches of fibers were internally equilibrated with cut end solutions containing two calcium indicators, antipyrylazo III (AP III) and fura-2, and different concentrations of free Mg2+ (25 microM-1 mM) obtained by adding appropriate total amounts of ATP and magnesium to the solutions. Changes in AP III absorbance were used to monitor [Ca2+] and [Mg2+] transients, whereas fura-2 fluorescence was mostly used to monitor resting [Ca2+]. Shortly after applying an internal solution containing less than 60 microM free Mg2+ to the cut ends of depolarized fibers most of the fibers exhibited spontaneous repetitive movements, suggesting that free internal Mg2+ might affect the activity of the sarcoplasmic reticulum (SR) calcium channels at rest. The spontaneous contractions generally subsided. In polarized fibers the maximal amplitude of the calcium transient elicited by a depolarizing pulse was about the same whatever the internal [Mg2+], but its decay after the end of the pulse slower in low [Mg2+]. In low [Mg2+] (less than 0.14 mM), the mean rate constant of decay obtained from fitting a single exponential plus a constant to the decay of the calcium transients was approximately 30% of its value in the control fibers (1 mM internal [Mg2+]). A model characterizing the main calcium removal properties of a frog skeletal muscle fiber, including the SR pump and the Ca-Mg sites on parvalbumin, was fitted to the decay of the calcium transients. Results of the fits show that in low internal [Mg2+] the slowing of the decay of the calcium transient can be well predicted by both a decreased rate of SR calcium uptake and an expected decreased resting magnesium occupancy of parvalbumin leading to a reduced contribution of parvalbumin to the overall rate of calcium removal. These results are thus consistent with the known properties of parvalbumin as a Ca-Mg buffer and furthermore suggest that in an intact portion of a muscle fiber, the activity of the SR calcium pump can be affected by the level of free Mg2+.


Low myoplasmic Mg2+ potentiates calcium release during depolarization of frog skeletal muscle fibers.

The role of intracellular free magnesium concentration ([Mg2+]) in modulating calcium release from the sarcoplasmic reticulum (SR) was studied in voltage-clamped frog cut skeletal muscle fibers equilibrated with cut end solutions containing two calcium indicators, fura-2 and antipyrylazo III (AP III), and various concentrations of free Mg2+ (25 microM-1 mM) obtained by adding appropriate total amounts of ATP and magnesium to the solutions. Changes in AP III absorbance were used to monitor calcium transients, whereas fura-2 fluorescence was used to monitor resting calcium. The rate of release (Rrel) of calcium from the SR was calculated from the calcium transient and found to be increased in low internal [Mg2+]. After correcting for effects of calcium depletion from the SR and normalization to SR content, the mean values of the inactivatable and noninactivatable components of Rrel were increased by 163 and 46%, respectively, in low Mg2+. Independent of normalization to SR content, the ratio of inactivatable to noninactivatable components of Rrel was increased in low internal [Mg2+]. Both observations suggest that internal [Mg2+] preferentially modulates the inactivatable component of Rrel, which is thought to be due to calcium-induced calcium release from the SR. This could also explain the observation that, in low internal [Mg2+], the time to the peak of the calcium transient for a 5-ms depolarizing pulse was not very different from the time to the peak of the delta [Ca2+] for a 10-ms pulse of the same amplitude. Finally, in low internal [Mg2+], the calcium transient elicited by a short depolarizing pulse was in some cases clearly followed by a very slow rise of calcium after the end of the pulse. The observed effects of reduced [Mg2+] on calcium release are consistent with a removal of the inhibition that the normal 1 mM myoplasmic [Mg2+] exerts on calcium release in skeletal muscle fibers.


Visual responses in teleosts. Electroretinograms, eye movements, and circadian rhythms.

We have recorded ocular potentials in response to brief flashes of light from two teleosts, the white perch (Roccus americana) and the green sunfish (Lepomis cyanellus). The animals were respired and maintained in an alert state for up to 2 d. Responses were recorded with corneal and transcleral electrodes. The responses of green sunfish were composed of electroretinogram (ERG) and eye movement potentials, whereas the responses in white perch contained only the ERG. Injection of curare abolished the sunfish eye movement potentials, unmasking the ERG. Observation under infrared illumination established a direct relationship between eye movements and the fast potentials which could be abolished by curare. We found no evidence of circadian changes in the amplitude of the ERG b-wave of either species. However, our results combined with those of a previous study of sunfish ocular potentials (Dearry, A., and B. Barlow, Jr. 1987. J. Gen. Physiol. 89: 745-770) suggest that the sunfish visual system exhibits rhythmic changes in oculomotor responses, which appear to be controlled by a circadian oscillator.


Modulation of voltage-dependent Ca channel current by arachidonic acid and other long-chain fatty acids in rabbit intestinal smooth muscle.

The effects of arachidonic acid (AA) and other long-chain fatty acids on voltage-dependent Ca channel current (ICa) were investigated, with the whole cell patch clamp method, in longitudinal smooth muscle cells of rabbit ileum. 10-30 microM AA caused a gradual depression of ICa. The inhibitory effect of AA was not prevented by indomethacin (10 microM) (an inhibitor of cyclooxygenase) or nordihydroguaiaretic acid (10 microM) (an inhibitor of lipoxygenase). 1-(5-Isoquinolinesulfonyl)-2-methylpiperazine (H7; 25-50 microM) or staurosporine (2 microM) (inhibitors of protein kinase C) did not block the AA-induced inhibition of ICa, and application of phorbol ester (a protein kinase C activator) (phorbol-12,13-dibutyrate, 0.2 microM) did not mimic the AA action. Some other cis-unsaturated fatty acids (palmitoleic, linoleic, and oleic acids) were also found to depress ICa, while a trans-unsaturated fatty acid (linolelaidic acid) and saturated fatty acids (capric, lauric, myristic, and palmitic acids) had no inhibitory effects on ICa. Myristic acid consistently increased the amplitude of ICa at negative membrane potentials. The present results suggest the possible role of AA, and perhaps other fatty acids, in the physiological and/or pathological modulation of ICa in smooth muscle.


Functional expression in Escherichia coli of proteins B and C from soluble methane monooxygenase of Methylococcus capsulatus (Bath).

Methylococcus capsulatus (Bath) uses a soluble methane monooxygenase (sMMO) to catalyse the oxidation of methane to methanol. sMMO is comprised of three components; A, B and C. Protein C (the reductase) transfers electrons from NADH to protein A (the hydroxylase) which contains the active site, and protein B regulates this electron flow. The five genes encoding the sMMO proteins and their subunits are clustered and have been cloned in Escherichia coli. A DNA fragment containing mmoB, the gene encoding protein B, was subcloned into pT7-5, a plasmid of the T7 RNA polymerase promoter expression system. Upon induction, E. coli expressed protein B which was fully functional after purification. The gene encoding protein C, mmoC, was amplified with unique restriction sites at each end using the polymerase chain reaction and then subcloned into pT7-7 (a plasmid similar to pT7-5 but containing its own ribosome-binding site and ATG start codon). Protein C expressed in E. coli was also found to be functional. This is the first report of the functional expression of methanotroph methane monooxygenase genes in a heterologous host and represents a significant step forward in our analysis of the assembly and catalysis of sMMO.


A monofunctional prephenate dehydrogenase created by cleavage of the 5' 109 bp of the tyrA gene from Erwinia herbicola.

A cohesive phylogenetic cluster that is limited to enteric bacteria and a few closely related genera possesses a bifunctional protein that is known as the T-protein and is encoded by tyrA. The T-protein carries catalytic domains for chorismate mutase and for cyclohexadienyl dehydrogenase. Cyclohexadienyl dehydrogenase can utilize prephenate or L-arogenate as alternative substrates. A portion of the tyr A gene cloned from Erwinia herbicola was deleted in vitro with exonuclease III and fused in-frame with a 5' portion of lacZ to yield a new gene, denoted tyrA*, in which 37 N-terminal amino acids of the T-protein are replaced by 18 amino acids encoded by the polycloning site/5' portion of the lacZ alpha-peptide of pUC19. The TyrA* protein retained dehydrogenase activity but lacked mutase activity, thus demonstrating the separability of the two catalytic domains. While the Km of the TyrA* dehydrogenase for NAD+ remained unaltered, the Km for prephenate was fourfold greater and the Vmax was almost twofold greater than observed for the parental T-protein dehydrogenase. Activity with L-arogenate, normally a relatively poor substrate, was reduced to a negligible level. The prephenate dehydrogenase activity encoded by tyrA* was hypersensitive to feedback inhibition by L-tyrosine (a competitive inhibitor with respect to prephenate), partly because the affinity for prephenate was reduced and partly because the Ki value for L-tyrosine was decreased from 66 microM to 14 microM. Thus, excision of a portion of the chorismate mutase domain is shown to result in multiple extra-domain effects upon the cyclohexadienyl dehydrogenase domain of the bifunctional protein.(ABSTRACT TRUNCATED AT 250 WORDS)


Lack of homology between two haloacetate dehalogenase genes encoded on a plasmid from Moraxella sp. strain B.

Two genes encoding haloacetate dehalogenases, H-1 and H-2, are closely linked on a plasmid from Moraxella sp. strain B. H-1 predominantly acts on fluoroacetate, but H-2 does not. To elucidate the molecular relationship between the two enzymes, we compared their structural genes. Two restriction fragments of the plasmid DNA were subcloned on M13 phages and their nucleotide sequences were determined. The sequence of each fragment contained an open reading frame that was identified as the structural gene for each of the two dehalogenases on the basis of the following criteria; N-terminal amino acid sequence, amino acid composition, and molecular mass. The genes for H-1 and H-2, designated dehH1 and dehH2, respectively, had different sizes (885 bp and 675 bp) and G+C contents (58.3% and 53.4%). Sequence analysis revealed no homology between the two genes. We concluded that the dehalogenases H-1 and H-2 have no enzyme-evolutionary relationship. The deduced amino acid sequence of the dehH1 gene showed significant similarity to those of three hydrolases of Pseudomonas putida and a haloalkane dehalogenase of Xanthobacter autotrophicus. The dehH2 coding region was sandwiched between two repeated sequences about 1.8 kb long, which might play a part in the frequent spontaneous deletion of dehH2 from the plasmid.


Molecular genetics of the extracellular lipase of Pseudomonas aeruginosa PAO1.

The structural gene (lipA) coding for the extracellular lipase of Pseudomonas aeruginosa PAO1 has been cloned on plasmid pSW118. Nucleotide sequence analysis revealed a gene of 936 bp. lipA codes for a proenzyme of 311 amino acids including a leader sequence of 26 amino acids. The mature protein was predicted to have a M(r) of 30134, an isoelectric point of 5.6, and a consensus sequence (IGHSHGG) typical of lipases. Furthermore it is highly homologous (greater than 60%) to other lipases from various pseudomonads. The lipA gene failed to hybridize detectably with genomic DNA from other Pseudomonas species except P. alcaligenes, even under relaxed stringency. Located 220 bp downstream of the lipA gene, is an open reading frame (ORF2, lipH) which encodes a hydrophilic protein (283 amino acids; M(r) 33587) that shows some homology to the limA gene product of P. cepacia. In complementation tests of lipase-defective mutants, lipH was shown to be necessary for expression of active extracellular lipase in P. aeruginosa PAO1.


A survey of Col plasmids in natural isolates of Escherichia coli and an investigation into the stability of Col-plasmid lineages.

A survey of colicins in the ECOR reference collection of Escherichia coli is presented. Twenty-five of the 72 ECOR strains exhibited a phenotype consistent with colicin production and E. coli isolated from human hosts were more likely to be colicinogenic than those from animal hosts. Multiple representatives of two Col plasmids, low-molecular-mass ColE1 plasmids and high-molecular-mass, conjugative ColIa plasmids were isolated from the ECOR collection and were examined with a combination of restriction fragment and Southern analysis. These data suggested that ColE1 plasmids comprise a stable (cohesive) plasmid lineage, while ColIa plasmids represent a family of distinct plasmid lineages united by the presence of the colicin Ia operon.


Cloning, sequencing and expression of the gene encoding the cell-envelope-associated proteinase from Lactobacillus paracasei subsp. paracasei NCDO 151.

The gene encoding the cell-envelope-associated proteinase of Lactobacillus paracasei subsp. paracasei NCDO 151 (formerly Lactobacillus casei NCDO 151) was cloned and sequenced. The gene was located on the chromosome and encoded a polypeptide of 1902 amino acids. The proteinase is N-terminally cleaved upon maturation. It shows extensive homology to the Lactococcus lactis subsp. cremoris Wg2 proteinase. Similar to the situation in Lactococcus, a maturation gene was found upstream of the proteinase gene. The cloned proteinase gene was expressed in Lactobacillus plantarum. However, no expression was observed when the gene was cloned in Lactococcus lactis.


Molecular cloning and characterization of an alkalophilic Bacillus sp. C125 gene homologous to Bacillus subtilis sec Y.

A 1.8 kb HindIII DNA fragment containing the secY gene of alkalophilic Bacillus sp. C125 has been cloned into plasmid pUC119 using the B. subtilis secY gene as a probe. The complete nucleotide sequence of the cloned DNA indicated that it contained one complete ORF and parts of two other ORFs. The similarity of these ORFs to the sequences of the B. subtilis proteins indicated that they were the genes for ribosomal protein L15-SecY-adenylate kinase, in that order. The gene product of the alkalophilic Bacillus sp. C125 secY homologue was composed of 431 amino acids and its M(r) value has been calculated to be 47,100. The distribution of hydrophobic amino acids in the gene product suggested that the protein was a membrane integrated protein with ten transmembrane segments. The total amino acid sequence of alkalophilic Bacillus sp. C125 secY homologue showed 69.7% homology with that of B. subtilis secY. Regions of remarkably high homology (78% identity) were present in transmembrane regions, and cytoplasmic domains (73% identity) with less homologous regions present in extracellular domains (43% identity).


Stable inheritance of shuttle vectors based on plasmid pIM13 in a mutant strain of Clostridium acetobutylicum.

New shuttle vectors for Clostridium acetobutylicum were constructed, using as replicons the Gram-positive plasmid pIM13, and derivatives of the Gram-negative plasmid pBR322, including pUC19. These vectors transformed C. acetobutylicum at a high frequency (up to 10(6) transformants per microgram DNA) by PEG-mediated protoplast transformation. A mutant host strain, NI-4082, was isolated on the basis of its ability to maintain plasmid pIM13 stably in the absence of selection pressure. The shuttle vectors showed no segregational or structural instability in this mutant strain. Moreover, the results suggested a relationship between segregational instability and the multimerization of pIM13 in C. acetobutylicum. The host/vector system described possessed all the properties required for efficient gene cloning in this species.


Isolation and characterization of a conjugative plasmid from Legionella pneumophila.

The conjugative properties of an indigenous 85 MDa plasmid (designated pCH1) from Legionella pneumophila were studied. To determine if pCH1 was transmissible by conjugation, mating experiments were performed between legionellae that harboured pCH1 and several plasmid-less recipients. Plasmid transfer was monitored by colony hybridization, using a cloned 21.0 kb SalI restriction fragment from pCH1 as a probe. The results from these experiments showed that pCH1 could be conjugatively transferred into several strains of L. pneumophila serogroup 1 but not into strain Bloomington-2 (serogroup 3) or Escherichia coli. Southern hybridization experiments in which pCH1 DNA was used as a probe showed that pCH1 does not share homology with other indigenous L. pneumophila plasmids. There was no detectable DNA homology between pCH1 and L. pneumophila chromosomal DNA. Additional mating experiments revealed that pCH1 was unable to mobilize the L. pneumophila chromosome. The conjugative transfer of pCH1 into plasmid-less avirulent or virulent serogroup 1 strains did not alter the intracellular growth characteristics of these strains in U937 cells, a human-monocyte-like cell line, or in the amoeba Hartmannella vermiformis. These results suggest that pCH1 does not contribute to the ability of L. pneumophila to enter or grow within eukaryotic cells.


Characterization of bacteriophage BFK20 from Brevibacterium flavum.

Bacteriophage BFK20 was isolated from a Brevibacterium flavum strain that had become contaminated during industrial fermentation. BFK20 has a polyhedral head 50 nm wide and a non-contractile tail 200 nm long and 10 nm in diameter. The genome of this bacteriophage consists of a linear double stranded DNA molecule of 44-45 kb with cohesive ends. The capsid of phage BFK20 contains nine polypeptides with molecular masses from 22.0-108.0 kDa. BFK20 DNA was used as a donor for fragments carrying promoters and transcription-terminators.


Molecular characterization of two bacteriophages isolated from Desulfovibrio vulgaris NCIMB 8303 (Hildenborough).

A preliminary endonuclease restriction map of a bacteriophage isolated from Desulfovibrio vulgaris has been established. BamHI cleaved whole phage DNA into four fragments while HindIII cut the same DNA into seven fragments. Mapping studies succeeded in linking the four BamHI fragments into two DNA segments; however, no linkage between the two segments was detected. These data imply that two phages were induced from cultures of D. vulgaris and that the two segments represented the DNA from these phages. Support for this hypothesis came from size approximation of restriction enzyme fragments, electron micrographs, and density gradients.


Molecular characterization of the genes encoding acetohydroxy acid synthase in the cyanobacterium Spirulina platensis.

The enzyme acetohydroxy acid synthase (AHS), which catalyses the first common step in the biosynthesis of isoleucine, leucine and valine, has been demonstrated to be present in Spirulina platensis in two isoenzymic forms. The complete nucleotide sequences of the genes ilvX and ilvW encoding these two enzymes have been determined. Sequence analysis revealed the presence of two open reading frames, of 1836 and 1737 nucleotides for ilvX and ilvW, respectively. The predicted amino acid sequences of the two isoenzymes, compared with the Synechococcus PCC 7942 AHS enzyme and the large subunits of the Escherichia coli AHSI, II, III isoenzymes, revealed a notable degree of similarity. A small subunit has not been identified for either of the S. platensis AHS isoenzymes. Analysis by Northern blot hybridization demonstrated that the ilvX and ilvW genes are transcribed to give mRNA species of approximately 2.15 kb and 1.95 kb, respectively.


Generation of genetic recombinants in Trichosporon cutaneum by spontaneous segregation of protoplast fusants.

Auxotrophic mutants were isolated in two strains of Trichosporon cutaneum. Complementing auxotrophs were hybridized by protoplast fusion. Some of the fusants were apparently transient diploids and segregated to give recombinant marker combinations.


Cloning and expression of multiple cellulase cDNAs from the anaerobic rumen fungus Neocallimastix patriciarum in Escherichia coli.

A cDNA expression library of the rumen fungus Neocallimastix patriciarum was made in Escherichia coli. Cellulolytic clones were identified by screening on a medium containing carboxymethylcellulose. Restriction mapping and Southern hybridization analysis of selected clones revealed three distinct cellulase cDNAs, designated celA, celB and celC. Studies on the substrate specificity showed that the enzyme encoded by celA had high activity towards amorphous and microcrystalline cellulose, while the celB and celC enzymes had relatively high activity on carboxymethylcellulose, with little activity on microcrystalline cellulose. Analysis of hydrolysis products from defined cellodextrins showed that the celB and celC enzymes hydrolysed beta-1,4-glucosidic linkages randomly, whereas the celA enzyme cleaved cellotetraose to cellobiose, and cellopentaose to cellobiose and cellotriose. Cellobiose was also the only product detectable from hydrolysis of microcrystalline cellulose by the celA enzyme. Based on substrate specificity and catalytic mode, celA appears to encode a cellobiohydrolase, while celB and celC encode endoglucanases. Northern blot hybridization analysis showed that expression of the three cellulase transcripts in N. patriciarum was induced by cellulose.


Regulation of restrictocin production in Aspergillus restrictus.

The production of restrictocin (a cytotoxin that specifically cleaves ribosomal RNA) by cultures of Aspergillus restrictus grown in liquid medium was investigated. The function of restrictocin, the method of its accumulation and the mode of resistance to restrictocin in A. restrictus are unknown. Previous studies have indicated that restrictocin accumulates in the medium with culture age. These observations have been extended in this study by cloning the cDNA of the res gene and using this cDNA clone to probe the onset of messenger RNA synthesis in the cells. The results of the Northern analysis were compared to the production and accumulation of restrictocin and morphological differentiation of the cells in culture. Restrictocin was found in the medium at the same time that mRNA was detected in the cells. This suggests that the leader sequence encoded by the cDNA provides an efficient secretion system for the protein. Both the protein and the mRNA were detected coincident with the formation of differentiated cell structures. These structures develop into conidiophores with one layer of sterigmata and conidia forming from the sterigmata. These results suggest that restrictocin is either involved in the process of conidiation or is coordinately regulated with differentiation leading to conidiation.


Production and localization of restrictocin in Aspergillus restrictus.

The production and secretion of restrictocin (a cytotoxin that cleaves ribosomal RNA) by cultures of the fungus Aspergillus restrictus was investigated. Previous studies have indicated that restrictocin production in liquid culture coincides with the appearance of differentiated cell structures. A study of the correlation between the appearance of differentiated structures and restrictocin production was conducted with A. restrictus grown on agar medium. Restrictocin was found to be associated with the cell mass of the agar-grown culture (in contrast to liquid cultures), and was first observed when aerial hyphae emerged. Restrictocin levels increased until the time of conidiation, after which they fell off sharply. No restrictocin could be found in the agar medium. The presence of restrictocin upon and within various cell structures was determined by immunofluorescent laser microscopy. This study showed that restrictocin became localized to the conidiophores and phialides during the process of conidiation. Prior to this, restrictocin was found within the hyphae in localized concentrations that may correspond to secretory vesicles.


Effect of nitrogen source on the levels of nitrate reductase in the yeast Hansenula anomala.

Levels of nitrate reductase (NR) protein in Hansenula anomala and Hansenula wingei were determined using specific antiserum raised against the enzyme from H. anomala. Extracts from nitrate-grown cells contained NR protein, while in those from cells grown on ammonium, glutamine or peptone, no cross-reacting material could be observed. Enzyme activity correlated with the levels of cross-reacting material. When nitrate was used as nitrogen source, NR was always present, even in cultures with ammonium, glutamine or peptone, although in these cases both the levels of activity and protein were lower. NR activity was consistently two to four times higher in cells grown in glucose than in cells grown in ethanol. Nitrate was required for NR induction, and deprivation of nitrate from nitrate-grown cells resulted in a rapid loss of NR activity.


Purification and characterization of L-2,4-diaminobutyrate decarboxylase from Acinetobacter calcoaceticus.

Acinetobacter calcoaceticus ATCC 23055 produces a large amount of 1,3-diaminopropane under normal growth conditions. The enzyme responsible, L-2,4-diaminobutyrate (DABA) decarboxylase (EC 4.1.1.-), was purified to electrophoretic homogeneity from this bacterium. The native enzyme had an M(r) of approximately 108,000, with a pI of 5.0, and was a dimer composed of identical or nearly identical subunits with apparent M(r) 53,000. The enzyme showed hyperbolic kinetics with a Km of 1.59 mM for DABA and 14.6 microM for pyridoxal 5'-phosphate as a coenzyme. The pH optimum was in the range 8.5-8.75, and Ca2+ gave a much higher enzyme activity than Mg2+ as a cationic cofactor. N-gamma-Acetyl-DABA, 2,3-diaminopropionic acid, ornithine and lysine were inert as substrates. The enzyme was different in subunit structure, N-terminal amino acid sequence and immunoreactivity from the DABA decarboxylase of Vibrio alginolyticus previously described.


The use of rRNA sequences and fluorescent probes to investigate the phylogenetic positions of the anaerobic ciliate Metopus palaeformis and its archaeobacterial endosymbiont.

The polymerase chain reaction (PCR) was used to amplify small-subunit ribosomal DNA from the anaerobic ciliated protozoon Metopus palaeformis, and from its uncultured endosymbiotic bacteria. This was accomplished directly from total DNA extracted from protozoa without prior isolation or enrichment for symbiont cells. The double-stranded amplification products were precipitated and directly sequenced using the linear PCR reaction. Fluorescent oligonucleotide probes were designed and used in whole-cell hybridizations to provide direct visual evidence that the sequences originated from the host ciliate and from the endosymbiont. Phylogenetic analysis of the Metopus palaeformis sequence consistently placed it as a deep-branching lineage near the root of the ciliate tree. However, the present data were insufficient to resolve the detailed relationship between Blepharisma and Metopus and thus to determine if the heterotrichs are mono- or paraphyletic. Phylogenetic analysis of the symbiont partial sequence clearly demonstrated that it is an archaeobacterium and that it is closely related to, but distinct from, Methanobacterium formicicum.


Identification of erythrocyte-binding antigens in Helicobacter pylori.

The surface antigens of Helicobacter pylori conferring erythrocyte-binding activity were obtained by adsorption onto formaldehyde-treated dog and goat erythrocytes from supernatant fractions of sonicated bacteria and elution using a high concentration of NaCl. The desorbed material was analysed by SDS-PAGE and immunoblotting with anti-whole-cell serum to agar-grown bacteria which had been absorbed with broth-grown, non-haemagglutinating cells (haemagglutination-associated antiserum). Two polypeptides with molecular masses of 25 and 59 kDa were revealed as erythrocyte-binding antigens. Strains which agglutinated both dog and goat erythrocytes possessed both these erythrocyte-binding antigens, whereas an antigenically cross-reactive 24 kDa polypeptide was present in a strain which only agglutinated goat erythrocytes. Haemagglutinin material was extracted from H. pylori using n-octylglucopyranoside and purified by Sepharose chromatography and sucrose density gradient ultracentrifugation. The purified extract directly agglutinated erythrocytes in a neuraminyl-lactose-sensitive and neuraminidase-sensitive manner. The 59 kDa polypeptide was not present in the purified haemagglutinin preparation. The haemagglutination-associated antiserum reacted strongly with the 25 kDa polypeptide band which was the most prominent polypeptide band on analysis of the purified haemagglutinin preparation by SDS-PAGE and silver staining. Thus, H. pylori possesses at least two adhesins, one of which recognises a N-acetylneuraminic acid (alpha 2-3) moiety of receptors, the other being of unknown receptor specificity. Differences in the antigenicity and molecular masses of these adhesins in individual strains may underlie differences in receptor-binding specificities and haemagglutination profiles.


Binding of purified Bacillus sphaericus binary toxin and its deletion derivatives to Culex quinquefasciatus gut: elucidation of functional binding domains.

Highly larvicidal strains of Bacillus sphaericus produce a binary toxin composed of 51 and 42 kDa proteins which binds to sharply delineated regions of the gastric caecum and posterior midgut of susceptible larvae of the mosquito Culex quinquefasciatus. To investigate the role of the individual subunits and the organization of functional binding regions within the toxin, plasmids were constructed for the expression in Escherichia coli of the toxin proteins and their NH2- and COOH-terminal deletion derivatives as fusions with glutathione S-transferase (GST). Toxin proteins were purified by affinity chromatography followed by cleavage from the GST carrier with thrombin. The LC50 values for the purified toxin proteins and their deletion derivatives were determined. The binding patterns of fluorescently labelled toxin suggested that the 51 kDa protein is the primary binding component of the toxin and mediates the regional binding and internalization of the 42 kDa protein. Examination of the toxin deletion derivatives revealed that the NH2-terminal region of the 51 kDa protein was required for binding to the larval gut, whilst the COOH-terminal region was responsible for interacting with the 42 kDa protein. Toxicity was strongly correlated with the subsequent internalization of the toxin, probably by endocytosis.


Lack of correlation between colony morphology and lipooligosaccharide content in the Mycobacterium tuberculosis complex.

Rough and smooth colony variants of the Mycobacterium tuberculosis complex were compared with respect to their composition in trehalose-containing glycolipid antigens in view of the results of a recent investigation suggesting that the chemical basis of rough and smooth colony morphology in mycobacteria may reside in the occurrence of lipooligosaccharides. A careful chemical characterization of the individual glycolipids of the selected strains allowed the identification of the major glycolipids. The comparative study of the glycolipid content of the smooth Canetti strain, its spontaneous rough variant, and 16 additional strains of M. tuberculosis, M. bovis and M. africanum showed that the presence of lipooligosaccharides was not related to the morphology of the colonies.


Verbal communication, nonverbal communication, and private speech in lower and middle socioeconomic status preschool children.

Three types of communication--verbal, nonverbal, and private speech--were investigated in 4- and 5-year-old children. Multiple analyses of variance (MANOVAs) without IQ controlled and multiple analyses of covariance (MANCOVAs) with IQ as a covariate were computed and followed by ANOVAs and ANCOVAs to determine the effects of sex, age, and socioeconomic status (SES) on each of these types of communication. The ANOVAs and the ANCOVAs yielded the same conclusions. Results indicated that age and SES, but not sex, influence the use of the three communication types. From age 4 to age 5, private speech decreased for middle SES children and remained the same for lower SES children. Although lower SES children had more nonverbal communication at both age levels than middle SES children, nonverbal communication decreased for middle SES children and increased for lower SES children between the ages of 4 and 5 years. Both lower and middle SES groups increased in verbal communication between the two ages. The middle SES 4-year-olds used more verbal communication than their lower SES counterparts, and the difference was maintained at age 5. Although IQ is related to verbal communication, statistically controlling for the effects of IQ did not change the conclusions.


The influence of age, gender, and having a younger sibling on children's knowledge about babies.

This article reports on two studies designed to measure 4- to 7-year-old children's knowledge about babies and its relationship to gender role development. In Study 1, children were asked several questions about babies and were given the Sex Role Learning Index (SERLI; Edelbrock & Sugawara, 1978). Girls provided more answers to one question, and children with younger siblings provided more answers to another. Overall, however, there were few relationships between knowledge about babies and gender role development. The second study measured knowledge about babies with two measures. The first measure asked children to identify foods that babies could eat and activities babies were capable of doing. The second measure asked children to identify the names and uses of certain objects. Older children scored higher on the foods and activities measure. Children who had younger siblings performed better on the objects measure. Gender and gender role development showed little relationship to knowledge about babies.


Visual and auditory orienting responses in preterm infants: a comparison of three cohorts over time.

Visual and auditory orienting responses were compared in three cohorts of preterm infants (N = 351) born between 1977 and 1987. The infants were divided into one group born between 25 and 34 weeks gestational age (n = 140) and one group born between 35 and 37 weeks gestational age (n = 211). Ratings were made of the infants' best orienting responses to a bull's-eye, rattle, bell, voice, and face plus voice combined. Analyses of covariance were computed to adjust for changes in perinatal variables between 1977 and 1987. For both groups, there were significant differences between cohorts on orienting to the bull's-eye, with a trend for the 25- to 34-week group on orienting to the rattle. Behavioral outcome for preterm infants has improved with changes in neonatal intensive care.


Effects of real gender and labeled gender on adults' perceptions of infants.

Three experiments were conducted to discover factors mediating adults' perceptions of male and female infants. In the first experiment, college students were shown 30-s videotapes of four male and four female babies, each of whom was randomly labeled with a male or a female name. Infants labeled as male were perceived as significantly more masculine and stronger than those labeled as female. Discriminant analyses revealed that both rated masculinity and the combination of ratings on male stereotyped traits differentiated infants labeled as male or female. Analyses of real gender revealed that boys were rated as less sensitive and stronger than girls. Discriminant analyses suggested that the combination of less sensitive, more of a problem, more mature, and more playful best differentiated real males from real females. In Experiment 2, the findings of Experiment 1 were confirmed with a sample of mothers of young infants. In Experiment 3 college students' judgments of the sex of the eight babies were correctly predicted from the sensitivity ratings of these babies in Experiment 1. It appears that there is a complex of cues from which adults make judgments of infants' gender and inferences about their characteristics: Boys may appear stronger, more playful, and more of a problem, and girls seem to look more sensitive. Implications for further studies of gender labeling and for sex typing are discussed.


Meaning in life and psychological well-being: an empirical study using the Chinese version of the Purpose in Life questionnaire.

The Chinese version of the Purpose in Life questionnaire (C-PIL) was administered to 2,150 Chinese secondary school students, along with other instruments assessing psychiatric symptoms and positive mental health. Total C-PIL and its two subscales, Quality of Existence (QEXIST) and Purpose of Existence (PEXIST), correlated significantly with all measures of psychological well-being. Relative to PEXIST scores, QEXIST scores were found to be more predictive of psychological well-being. Subjects with different existential status (defined by high vs. low levels of QEXIST and PEXIST) were associated with different degrees of psychological well-being. The concept of meaning in life, as indexed by the C-PIL, and the hypothesis that life meaning is related to psychological well-being require further elaboration and refinement.


Parental conflict and male adolescent problem behavior.

Mothers in 30 clinical and 35 nonclinical families completed a behavior checklist for a target son and all other sons in the family who were between 13 and 17 years old. Each of these male adolescents completed two perceived parental conflict questionnaires for their biological parents. In each group, child behavior problems and parental conflict were correlated under two conditions: (a) using information pertaining to the target adolescent and (b) using information pertaining to the male adolescent experiencing the greatest behavior difficulties. Correlations increased in the nonclinical group when the information used was for the adolescent experiencing the greatest behavior difficulties.


Girls' identity formation in the changing social structure of Sri Lanka.

Adolescent girls (N = 118) from diverse social backgrounds in Sri Lanka described their views of womanhood. Qualities of the ideal woman considered most important were kindness and honesty, liking children, intelligence, and having a good job. Although the traditional role of the Sri Lankan woman is that of homemaker, most girls in the study (55%) drew the ideal woman working outside the home, often as a teacher or a doctor. Nevertheless, adolescent girls emphasized traditional qualities of the woman at both work and home as self-sacrificing and serving others.


Human absence epilepsies.

A historical review of the concept of absence seizures. Their clinical features are very suggestive but a diagnosis made solely on clinical grounds is not always safe. Comparable pitfalls exist in the interpretation of EEG patterns. Absence seizures belong to several epileptic syndromes. They are briefly described.


Mapping of cerebral energy metabolism in rats with genetic generalized nonconvulsive epilepsy.

The quantitative 2-[14C]deoxyglucose autoradiographic method was applied to measure local cerebral metabolic rates of glucose (LCMRglc) in a model of genetic petit-mal-like seizures in a strain of Wistar rats. During the experimental period, epileptic rats exhibited synchronous spike-and-wave discharges, whereas the EEG pattern of control animals was normal. Overall, LCMRglc was consistently higher in epileptic rats than in the non-epileptic controls. The increase in LCMRglc was widespread and concerned all cerebral functional systems studied, whether they exhibit spike-and-wave discharges (neocortex and thalamus), or not (limbic system). These results are in good accordance with positron-emission tomography measurements in humans with typical childhood absence epilepsy. There appears to be a lack of anatomical correlation between areas demonstrating hypermetabolism and areas where spike-and-wave discharges are recorded. The administration of 200 mg/kg ethosuximide completely suppressed spike-and-wave discharges in epileptic rats and did not change the EEG pattern in controls. However, LCMRglc were increased to the same extent over control values in epileptic rats whether they were injected with ethosuximide or untreated. By contrast, when epileptic rats were given 2 mg/kg haloperidol, the frequency and the length of spike-and-wave discharges increased, inducing almost a permanent petit-mal status epilepticus. Haloperidol did not change EEG pattern in controls. In haloperidol-treated epileptic rats, LCMRglc decreased to levels comparable to those measured in untreated control rats. In the presence of haloperidol, LCMRglc were similar in both control and epileptic rats. Thus, the diffuse increase in cerebral energy metabolism in epileptic rats as compared to controls is not directly related to the occurrence of spike-and-wave discharges, and may rather be associated with inhibitory mechanisms involved in their termination and suppression, as well as their spread to limbic and motor structures.


The tottering mouse: a critical review of its usefulness in the study of the neuronal mechanisms underlying epilepsy.

The tottering mouse resulted from a recessively inherited, autosomal, single-locus mutation which produces a very characteristic neurological and cellular phenotype. Almost simultaneously and late in the development of this mutant appears a triad of symptoms: frequent episodes of absence seizures with spike-and-wave discharges; more rarely occurring episodes of focal motor seizures; and ataxia. Electrographic, behavioural and pharmacological similarities to absence epilepsy in man make the tottering mouse a useful animal model for testing new anti-absence drugs. It also affords a unique opportunity to study the effects of multiple alleles on epileptic behaviour. The neuronal mechanisms underlying the generation of absence seizures in this mutant are apparently a combination of a generalized noradrenergic hyperactivity in the brain and some gene-linked, but unknown, conditions prevailing in an earlier phase of development at specific brain areas which induce the generalized forebrain hyper-innervation by locus coeruleus terminals. Several biochemically, microscopically and electrophysiologically identified cellular differences between normal and tottering mice are potential aspects of this primary developmental defect. Research into these gene-linked neuronal characteristics co-inherited with seizures in this mutant makes the tottering mouse a powerful tool in the study of cellular mechanisms underlying genetically determined factors in epileptogenesis.


Genetic absence epilepsy in rats from Strasbourg--a review.

We have selected a strain of rats and designated it the Genetic Absence Epilepsy Rat from Strasbourg (GAERS). In this strain, 100% of the animals present recurrent generalized non-convulsive seizures characterized by bilateral and synchronous spike-and-wave discharges accompanied with behavioural arrest, staring and sometimes twitching of the vibrissae. Spontaneous SWD (7-11 cps, 300-1,000 microV, 0.5-75 sec) start and end abruptly on a normal background EEG. They usually occur at a mean frequency of 1.5 per min when the animals are in a state of quiet wakefulness. Drugs effective against absence seizures in humans (ethosuccimide, trimethadione, valproate, benzodiazepines) suppress the SWD dose-dependently, whereas drugs specific for convulsive or focal seizures (carbamazepine, phenytoin) are ineffective. SWD are increased by epileptogenic drugs inducing petit mal-like seizures, such as pentylenetetrazol, gamma-hydroxybutyrate, THIP and penicillin. Depth EEG recordings and lesion experiments show that SWD in GAERs depend on cortical and thalamic structures with a possible rhythmic triggering by the lateral thalamus. Most neurotransmitters are involved in the control of SWD (dopamine, noradrenaline, NMDA, acetylcholine), but GABA and gamma-hydroxybutyrate (GHB) seem to play a critical role. SWD are genetically determined with an autosomal dominant inheritance. The variable expression of SWD in offsprings from GAERS x control reciprocal crosses may be due to the existence of multiple genes. Neurophysiological, behavioural, pharmacological and genetic studies demonstrate that spontaneous SWD in GAERS fulfill all the requirements for an experimental model of absence epilepsy. As the mechanisms underlying absence epilepsy in humans are still unknown, the analysis of the genetic thalamocortical dysfunction in GAERS may be fruitful in investigations of the pathogenesis of generalized non-convulsive seizures.


Cortical and thalamic lesions in rats with genetic absence epilepsy.

In generalized, non-convulsive, absence epilepsy, spike-and-wave discharges (SWD) are recorded in both the cortex and the thalamus. The effect of various cortical and thalamic lesions on the occurrence of spontaneous SWD was examined in rats from a strain with genetic absence epilepsy. Cortical ablations suppressed SWD recorded in the thalamus. KCl induced unilateral cortical spreading depression and transiently suppressed SWD in the ipsilateral cortex and thalamus; SWD recovered simultaneously in both structures. Bilateral thalamic lesions of the anterior nuclei, the ventromedial nuclei, the posterior area, or lesion of the midline nuclei did not suppress cortical SWD. However, large lesions of the lateral thalamus, including the specific relay and reticular nuclei, definitely suppressed ipsilateral SWD, and pentylenetetrazol, THIP or gammabutyrolactone failed to restore the cortical SWD. These results demonstrate that the neocortex and the specific thalamic nuclei are both necessarily involved in the generation of SWD in absence epilepsy.


Role of the thalamic reticular nucleus in the generation of rhythmic thalamo-cortical activities subserving spike and waves.

The role of the reticular thalamic nucleus (RTN) in pacing rhythmic cortical activities subserving spike-waves (SW) discharges has been investigated in rats. Intracellular recordings from thalamic slices in vitro demonstrated that RTN neurons from control animals possess a set of Ca2+/K+ membrane conductances which enable them to produce rhythmic oscillatory activities. In vivo, studies of Ca(2+)-conductance blockade by intrathalamic injections of Cd2+ were performed on 24 callosotomized Wistar rats displaying spontaneous SW discharges, bred at the Centre de Neurochimie, Strasbourg. A significant decrement in ipsilateral SW activity was consistently observed in all RTN-injected animals 40 min after Cd2+ injection. By contrast, animals which received Cd2+ injection into the ventroposterior complex (VP) showed only small changes in ipsilateral SW. It is concluded that Ca(2+)-dependent oscillatory properties of the RTN are critical for the expression of genetically determined SW discharges in the Wistar model.


Responses to N-methyl-D-aspartate are enhanced in rats with petit mal-like seizures.

The responses to the glutamate agonist N-methyl-D-aspartate (NMDA) were studied in the sensori-motor cortex of rats with petit mal-like seizures. In a first study, the changes in extracellular concentration of calcium elicited through ionophoretic application of NMDA at various depths in the cortex were measured in vivo. The results show that in the cortex of epileptic rats the NMDA responses are much more widely distributed than in the cortex of control rats. In a second study, a current-source density analysis of the responses elicited through electrical stimulation of the white matter was performed in slices of neocortex in vitro. These findings show that the NMDA-dependent component of the synaptic responses are more widely distributed and of longer duration in the cortex of epileptic rats than in that of control rats. Taken together, these results suggest that in this model of absence epilepsy NMDA-dependent mechanisms are important in the triggering and maintenance of epileptic activity.


Antitumor agents, 127. Bruceoside C, a new cytotoxic quassinoid glucoside, and related compounds from Brucea javanica.

Bruceoside C [5], a new quassinoid glucoside, and related compounds were isolated from Brucea javanica, and their structures were elucidated by spectral data. Bruceoside C showed potent cytotoxicities against KB, A-549, RPMI, and TE-671 tumor cells.


Transformations of (+-)-salsolinol into optically active O- and/or N-methylated derivatives by several Papaveraceae plants and their tissue-cultured cells.

(+-)-Salsolinol [3], a substance possibly inducing parkinsonism, was biotransformed into optically active O- and/or N-methylated salsolinols 5, 6, and 7 by several Papaveraceae plants and tissue-culture cells derived from these plants. The bioconversion of racemic salsolinol into optically active tetrahydroisoquinolines has never been shown to occur, either in animals or in plants.


A new triterpenoidal saponin from Acacia auriculiformis.

A new triterpenoidal saponin has been isolated from an aqueous EtOH extract of the legumes of Acacia auriculiformis and characterized as 3-O-([beta-D-xylopyranosyl(1----3)-beta-D-xylopyranosyl(1----4)-alpha-L- rhamnopyranosyl(1----2)]-[alpha-L-rhamnopyranosyl(1----4)]-beta-D- glucopyranosyl)-3,16,21-trihydroxyolean-12-en-28-oic acid [1] by chemical studies and spectral data.


An acetylated bidesmosidic saponin from Schefflera octophylla.

A new acetylated bidesmosidic triterpenoid saponin has been isolated from the leaves of Schefflera octophylla and structurally elucidated as 3-epi-betulinic acid 3-O-beta-D-6'-acetylglucopyranoside 28-[alpha-L-rhamnopyranosyl(1----4)-O-beta-D-glucopyranosyl(1----6)]-bet a-D-0-glucopyranoside [1].


Brominated beta-carbolines from the marine tunicate Eudistoma album.

Chemical investigation of the cytotoxic EtOH extract of the marine tunicate Eudistoma album led to the isolation of three brominated beta-carbolines which were characterized by their spectral data. Two of them, eudistomin E, which is already known for its potent antiviral activity, and the novel compound eudistalbin A [1], were shown to possess cytotoxic activity (ED50 less than 5.0 ng and 3.2 micrograms/ml respectively) in vitro against the growth of KB human buccal carcinoma cells. The third compound, eudistalbin B [2], is a new inactive natural product.


Second European Stroke Prevention Study. ESPS-2 Working Group.

The computerized design, aims, organization, and structure of the European Stroke Prevention Study 2 (ESPS-2) are presented. This clinical trial will investigate the effect of single and combination regimens of low-dose acetylsalicylic acid (50 mg daily) and dipyridamole (400 mg daily) versus placebo in secondary prevention of stroke and death.


McLeod syndrome: a distinct form of neuroacanthocytosis. Report of two cases and literature review with emphasis on neuromuscular manifestations.

McLeod syndrome was originally described on the basis of a specific blood group phenotype with weak expression of Kell antigens. This erythrocyte abnormality also causes acanthocytosis. The haematological findings are associated with abnormalities in other organ systems, including neuromuscular manifestations. A 51-year-old patient was followed up for 11 years. He presented with persistent muscle creatine kinase elevation and progressive heart disease and later developed a slowly progressive neuropathy and choreic movements. His younger brother presented with grand mal seizures, involuntary movements and high muscle creatine kinase when aged 43 years. Clinical myopathy was absent in both, yet muscle biopsy showed mild myopathic changes. The presence of a motor axonopathy was supported by electrophysiological findings. One brother also showed sensory axonopathy. The movement disorder suggested accompanying basal ganglia dysfunction. Earlier reports of McLeod syndrome are reviewed with respect to neuromuscular involvement. Absence of the Kx membrane protein seems to be the cause of this multi-system disorder.


Ataxic variant of adrenoleukodystrophy: MRI and CT findings.

A 28-year-old man developed slowly progressive dysarthria and gait disturbance over 7 years. Neurological examination revealed marked ataxia of articulation and gait, mild ataxia and spasticity of all four limbs without intellectual, visual, auditory, sensory or sphincter dysfunction. No physical signs of adrenal hypofunction were found. However, analysis of fatty acid of plasma sphingomyelin showed an increase in very long chain fatty acids, compatible with the diagnosis of adrenoleukodystrophy. Computed tomography disclosed marked atrophy of the cerebellum and pons, and bilateral low-density lesions in the deep while matter of the cerebellum. Magnetic resonance imaging showed these lesions more clearly, as well as other lesions in the middle and superior cerebellar peduncles, despite the absence of cerebral white matter involvement at the time of presentation. This is a rare case of adrenoleukodystrophy presenting as spinocerebellar degeneration and showing marked atrophy and several parenchymatous lesions of the cerebellum and brain stem.


Specific antigen binding by activated cerebrospinal fluid B lymphocytes in acute neuroborreliosis.

A recently developed immunocytochemical antigen-binding sandwich test for the identification of specific activated B lymphocytes was applied to cerebrospinal fluid cells and peripheral blood mononuclear cells in patients with acute neuroborreliosis. Discrimination of antigen-binding phagocytes was achieved by double staining with monoclonal antibodies. Specific activated B lymphocytes were much more numerous in cerebrospinal fluid than in blood, showing great interindividual differences. When intrathecal immunoglobulin production was present, the number of specific activated B lymphocytes was also high. The specificity of all activated B lymphocytes ranged from 10% to 60% and was higher in the acute stage than after treatment.


Extracranial vertebral artery dissection: nine cases.

Nine patients (six men, three women) with extracranial vertebral artery dissection are described. Their mean age was 39.1 years (range 17-66). In four cases dissection was "spontaneous"; in the other five cases there was a history of trivial trauma. Three patients had fibromuscular dysplasia, two were migraineurs, one had elastorrhexis. Treatment varied. Six received heparin, three acetylsalicylic acid or ticlopidine. Eight had good recovery. Two patients experienced recurrence when stopping acetylsalicylic acid. The pathogenesis of dissections and the distinction between spontaneous dissections and those associated with minor trauma are discussed.


Multimodal tests of cerebrovascular reactivity in migraine: a transcranial Doppler study.

Altered cerebral vasoreactivity (CVR) has been implicated in migraine. To test this hypothesis, we studied CVR as measured by transcranial Doppler ultrasound (TCD) in 11 migraineurs and 12 healthy controls of similar age. Mean flow velocities (MFV) in the middle cerebral artery (MCA) were recorded during a cognitive and two motor tasks. MFV in the posterior cerebral artery (PCA) were measured during photic stimulation and observation of complex images. The increase of MFV in the MCA during the cognitive task was greater in migraineurs than in controls (9.1% vs 5.0%; P = 0.06). The increase of MFV in both tests for PCA reactivity was significantly greater in migraineurs than in controls: 17.4% vs 9.9% for photic stimulation (P less than 0.05) and 20.3% vs 10.2% for observation of complex images (P less than 0.05). Owing to overlap of individual results, the discriminative value of both tests was unsatisfactory. The variability of flow velocities as measured by standard deviations of MFV was significantly greater in migraineurs than in controls during all tests of PCA vasoreactivity. Differences in CVR between migraineurs and normal controls may be detected by TCD testing, in particular in the PCA territory. For individual diagnostic purposes, CVR tests proved to be insufficient.


Thalamic ataxia.

Seventeen patients with hemiataxia as a manifestation of thalamic infarction were studied. Hemiataxia had the main clinical characteristics of a "cerebellar type" of ataxia, though it never occurred in isolation, being associated with ipsilateral sensory disturbance (hemiataxia-hypaesthesia) in 7 patients, with ipsilateral sensory disturbance and hemiparesis (hypaesthetic ataxic hemiparesis) in 8 patients, and with hemiparesis (ataxic hemiparesis) in 2 patients. Recovery was good, and in all patients the sensory and motor disturbances improved or cleared before the hemiataxia. All patients had an infarct involving the lateral part of the thalamus (thalamogeniculate territory in 16, tuberothalamic territory in 1), also affecting the posterior limb of the internal capsule (PLIC) in 7 patients. Hemiataxia seemed linked to involvement of the caudal part of the ventral lateral nucleus of the thalamus or the immediately adjacent medial part of the PLIC. These structures are near the corticospinal pathways and the ventral posterior nucleus of the thalamus, explaining why hemiataxia is associated with hemiparesis or hypaesthesia in this type of infarct.


Familial CNS tumors.

A family with central nervous system tumors in five individuals in two generations is reported. Of five sisters, two had an isolated meningioma and one an ependymoma. Amongst the four children of those affected there was a son with two cerebral meningiomas and a daughter with a spinal meningioma. Signs typical of von Recklinghausen neurofibromatosis (NF1) or bilateral acoustic neurofibromatosis (NF2) were not present. The pattern is consistent with autosomal dominant inheritance with relatively high penetrance.


Langerhans' cell histiocytosis and the nervous system.

We report two cases of Langerhans' cell histiocytosis with unusual central nervous system (CNS) involvement. The first patient had behavioural disturbances, memory loss and diabetes insipidus. His response to a range of treatments was poor. The second patient presented with seizures and headaches suggestive of raised intracranial pressure. Etoposide (VP16) chemotherapy led to a dramatic clinical and radiological improvement. The various CNS manifestations of Langerhans' cell histiocytosis and their management are discussed.


Prevalence of hereditary ataxias and spastic paraplegias in Molise, a region of Italy.

An epidemiological survey of hereditary ataxias and paraplegias was conducted in Molise, a region of Italy (335, 211 inhabitants on 1 January 1989). Total prevalence was 7.5 x 10(-5) inhabitants (95% confidence limits 4.8-11.1). There were 7 patients with Friedreich's disease, 5 with early onset cerebellar ataxia with retained tendon reflexes, 4 with ataxia-telangiectasia, 9 with hereditary spastic paraplegias (2 autosomal dominant and 7 autosomal recessive cases). There was no patient with autosomal dominant cerebellar ataxia.


HIV disease and levels of prevention.

It appears that several cofactors are involved in determining the pathogenesis of HIV disease. Exposure cofactors affect acquisition of HIV infection. Trigger cofactors determine a person's likelihood of being infected after exposure to HIV, or contribute to HIV disease progression in those who have been infected. Identification of the most common exposure and trigger cofactors should become a major focus of community health nursing practice at all levels of prevention. Changes in behaviors, early intervention activities, and rehabilitative measures related to known cofactors may improve the health of persons at all stages of HIV disease.


A collaborative effort between a state migrant health program and a baccalaureate nursing program.

This article reports on the way in which a unique collaborative effort between a service agency and a baccalaureate nursing program grew out of the needs of the two separate agencies. The first agency, the Colorado Migrant Health Program, was faced with a budget that was inadequate to meet the health-care needs of the migrant population. The second, a baccalaureate nursing program at the University of Colorado, was finding it increasingly difficult to locate appropriate generalized community health clinical placements with an opportunity for student discovery learning related to transcultural health care. Discussions of a collaborative effort ensued and it became evident that each had something to gain from such an effort. Now, after almost 10 years of cooperation, the commitment to the endeavor on the part of both agencies remains strong as the benefits to each continue to increase. The ultimate beneficiaries, however, are the migrant families receiving care and the students providing care. The way in which the program has evolved over time, a description of the current program, and a discussion of the resources that help to maintain the program are presented. In addition, the curriculum course design, and teaching strategy modifications necessary to facilitate the collaborative program are discussed.


Cultural impact of health-care access: challenges for improving the health of African Americans.

Disparities in health status of African Americans continue to exist. These disparities of poor health, in part, are attributed to decreased access to health-care services. However, culture plays a key role in health-care utilization patterns among African Americans. The purpose of this article is to examine cultural factors that affect health-care practices among African Americans and to identify specific community health nursing interventions that integrate these factors into health-care plans for African-American families and communities.


Predictors of the home environment of high-risk infants.

The quality of the home environment is an important predictor of the cognitive and social development of high-risk infants. Community health nurses (CHNs) have played a central role in the assessment and care of families with high-risk infants. We examined predictors of the home environment in a sample of 106 infants discharged from a neonatal intensive care unit. The most consistent predictor of an optimal home environment was an internal parental locus of control. Socioeconomic status affected dimensions of parenting related to cognitive stimulation but not emotional responsivity. Young mothers and those with other children are less responsive to their infants. Males are treated in a more responsive manner than females. These findings suggest that CHNs need to adopt an empowerment model of intervention and focus on the particular needs of young mothers and those with other children.


Intervention program for obese school children.

The purpose of this study was to implement and evaluate a program for obese school children. A pretest-posttest design was utilized. Data was collected related to weight status, skinfold measurements, self-esteem, and nutritional knowledge. A convenience sample of 26 children, in the fourth to sixth grades, completed this 9-week program. The results indicate that self-esteem increased significantly (p less than .001) between the pretest and posttest interval. Weight status and nutritional knowledge showed no improvement. Exercise was difficult to assess on self-report, therefore no conclusions were were drawn in relation to this variable. Future research will be directed toward refining this intervention program.


Nutrient intake as a time signal for circadian rhythm.

In considering nutrition and circadian rhythms, time-of-eating behavior is an inherited, genetically controlled pattern that can be phase-shifted by conditioning or training. In addition, there are metabolic responses to meal eating, such as entrainment of a number of enzyme levels, hormone concentrations and other metabolic and physiologic activities. To separate truly inherent rhythms from entrained responses, it is necessary to determine which oscillations continue their circadian rhythms under completely free-running conditions of no zeitgeber, such as darkness vs. light or meal eating vs. fasting. Where meal eating provides a time signal to a biological clock experiment, healthy animals-not starving but constantly receiving nutrition to avoid an eating time signal-should be used. The ability to determine which daily rhythms are not genetically inherent, but rather are responses to a particular time signal, requires the elimination of all but this time signal. By using free-running conditions that are totally without time-giving signals, entrainment by (response to) meal feeding is readily separated from inherited endogenous circadian rhythm. Both types of daily rhythm exist and both involve the biological clock mechanisms . A method by which to identify each is proposed.


Postnatal selenium repletion protects lungs of neonatal rats from hyperoxia.

We reported previously that Se-adequate neonatal rat pups born to Se-adequate dams were resistant to lung damage by hyperoxia. To assess whether early postnatal Se repletion could also protect developing pups reared under hyperoxia, female Sprague-Dawley rats (n = 20) were bred and fed a Se-deficient (0.04 microgram/g) diet during pregnancy. On d 1 postpartum, dams were divided into two groups and fed either a Se-deficient diet or a Se-repleted (0.5 microgram/g) diet. On d 4 postpartum, litters in each group were randomly assigned to either air or high oxygen (greater than 95% O2) environments. Histologic evaluation of lungs from d-8 pups indicated that Se repletion significantly reduced the incidence of lung lesions caused by hyperoxia. Selenium-repleted pups also had significantly greater lung volumes and internal surface areas. The 7-d period of Se repletion resulted in significantly elevated maternal milk Se concentrations compared with a Se-deficient group, which was reflected in the pups by elevated plasma and hepatic Se concentrations and Se-dependent glutathione peroxidase (SeGPx) activities. Pulmonary glutathione concentration and SeGPx activity in pups were affected by oxygen exposure only, not by Se nutrition. Therefore, early postnatal Se repletion can protect the developing lung from oxygen-induced injury, a protection that is not entirely due to the effects of Se on pulmonary SeGPx activity and glutathione concentration.


Dietary beta-carotene reduces serum lipid concentrations in spontaneously hypertensive rats fed a vitamin A-fortified and cholesterol-enriched diet.

The effects of dietary beta-carotene on serum lipid concentrations were examined in spontaneously hypertensive (SH) rats. Groups of SH rats were fed a semipurified, vitamin A-fortified and cholesterol-enriched diet supplemented with 0, 125, 250 or 500 mg beta-carotene/kg diet for a period of 44 d. beta-Carotene supplementation resulted in significant dose-related decreases in serum total, LDL and HDL cholesterol concentrations and serum total, VLDL and LDL triacylglycerol concentrations. The ratio of HDL cholesterol to total cholesterol was unchanged or slightly increased by dietary beta-carotene. The study suggests that dietary beta-carotene has antihyperlipidemic effects in SH rats. The effects in humans and the mechanism of the effects remain to be investigated.


Tryptophan intake influences infants' sleep latency.

Formula-fed infants have depressed plasma tryptophan concentration relative to human milk-fed infants. Because tryptophan alters sleep-waking patterns in adults, a study was designed to determine whether additional dietary tryptophan could elevate plasma tryptophan concentrations of formula-fed infants to concentrations characteristic of human milk feeding and whether differences in plasma tryptophan or the ratio of plama tryptophan to the sum of the other plasma large neutral amino acids (tryptophan:LNAA) were of behavioral significance. Infants were fed a formula (13 g protein/L; whey:casein, 34:66) containing either 0, 294, 588 or 882 mumol/L of added tryptophan. Infants fed human milk or commercial formula (15 g protein/L; whey:casein, 18:82) were included for comparison. In formula-fed groups, plasma tryptophan was directly related to tryptophan intake (r = 0.46, P less than 0.0005). Infants fed commercial formula or the formula without added tryptophan had lower (P less than 0.001) plasma tryptophan compared with infants fed human milk. Only the infants fed the highest tryptophan formula had significantly higher plasma tryptophan:LNAA ratios than the other experimental groups, and these ratios were similar to those of infants fed human milk. The plasma tryptophan:LNAA ratios, not plasma tryptophan concentrations, were predictive of differences in the infants' sleep latency; infants fed formula containing the highest tryptophan had sleep latencies of 18.7 min, significantly shorter (P less than 0.05) than those of infants fed formulas containing less added tryptophan (27.7 min). Feeding infants formulas differing in tryptophan concentration produced differences in sleep latency, which could influence neurobehavioral developments.(ABSTRACT TRUNCATED AT 250 WORDS)


Correlations of vitamin A and E intakes with the plasma concentrations of carotenoids and tocopherols among American men and women.

The authors investigated the association of diet and other factors with the plasma concentrations of carotenoids, retinol and tocopherols in a sample of 121 men and 186 women participating in two prospective investigations of dietary etiologies of chronic diseases. Lycopene (mean concentration, 0.82 mumol/L in men, 0.76 mumol/L in women), beta-carotene (mean 0.46 mumol/L in men, 0.58 mumol/L in women) and lutein (mean 0.28 mumol/L in men, 0.27 mumol/L in women) were the major circulating carotenoids. Among nonsmokers, dietary carotenoid, as typically calculated in epidemiologic studies, was significantly correlated with plasma beta-carotene (r = 0.34 in men, r = 0.30 in women), alpha-carotene (r = 0.52 in men, r = 0.37 in women) and lutein (r = 0.36 in men, r = 0.19 in women), but not with plasma zeaxanthin (r = 0.11 and r = 0.02) or lycopene (r = 0.13 and r = 0.01) after adjusting for plasma cholesterol and triglycerides, body mass index and energy intake. Total vitamin E intake was positively associated with plasma concentrations of alpha-tocopherol (r = 0.51 in men, r = 0.41 in women) and inversely associated with plasma concentrations of gamma-tocopherol (r = -0.51 in men r = -0.42 in women), but this was primarily due to use of vitamin E supplements. Measurements of specific carotenoids can provide independent information beyond the usual calculation of carotene intake in epidemiologic studies.


Adipocyte insulin responsiveness in female Sprague-Dawley rats fed a low fat diet containing a fat-mimetic carbohydrate.

Two experiments examined the effects of replacing high fat with low fat diets on adipocyte insulin sensitivity and response. Female Sprague-Dawley rats had free access to diets containing 21% (control), 61% (high fat) or 2% (low fat) of energy as fat. In the low fat diet a carbohydrate-based fat-mimetic carbohydrate replaced all but the essential fat present in the high fat diet. Insulin-stimulated glucose utilization by isolated adipocytes was measured after 10, 30 or 50 d. In a second study adipocyte insulin saturation curves were measured after 36 d. Rats fed the high fat diet for 30 d were insulin resistant and adipocyte basal and insulin-stimulated glucose utilization were depressed. The low fat diet initially stimulated glucose utilization of adipocytes but did not change insulin responsiveness. After 50 d there was no difference in glucose utilization between adipocytes from rats fed control and low fat diets. Insulin resistance in rats fed the high fat diet was associated with a nonsignificant reduction in insulin receptor number. These observations do not exclude the possibility of a post-receptor defect in glucose utilization.


Insulin insensitivity is rapidly reversed in rats by reducing dietary fat from 40 to 30% of energy.

The objective of this study was to determine the effect of reducing dietary fat content from 40 to 30% of total energy on body composition and insulin sensitivity in rats. Female Sprague-Dawley rats were fed either a control diet (30% energy as fat) or a high fat diet (40% energy as fat) for 10 wk. In vivo glucose tolerance tests indicated that both basal and glucose-stimulated insulin concentrations were elevated in rats fed the high fat diet. Two groups of rats offered the high fat diet were then fed the control diet (HF,C) and two that had received the high fat diet were then fed a diet (30% energy as fat) containing a fat-mimetic carbohydrate (HF,M). Two groups continued to receive the high fat diet and two groups continued to receive the control diet. Rats were killed 1 and 2 wk after the diet switch. In vivo glucose tolerance tests indicated that insulin sensitivity was corrected in HF,C and HF,M rats after 3 d. Body fat content was greater in rats fed the high fat diet and remained high after 7 d of consuming either diet with 30% of energy from fat. There was no effect of diet on liver fatty acid utilization or on diaphragm glucose oxidation. Adipocytes from all treatment groups were insulin resistant, possibly due to feeding status or age of the rats. The results of this study suggest that a moderate reduction in fat intake, from 40 to 30% of energy, can produce a rapid improvement of insulin sensitivity in insulin-insensitive rats, independent of changes in body fat content and irrespective of the means used to reduce dietary fat content.


Total parenteral nutrition with short- and long-chain triglycerides: triacetin improves nitrogen balance in rats.

Little is known about the long-term metabolic effects of parenteral administration of short-chain triglycerides. These studies were undertaken to investigate triacetin, the water-soluble triglyceride of acetate when it is incorporated into nutritionally balanced total parenteral nutrition formulas. Male Sprague-Dawley rats (n = 22) were fed an isovolemic, isocaloric and isonitrogenous diet for 7 d. The lipid energy represented 30% of the nonprotein energy with short-chain triglycerides representing 0, 50 or 90% of the lipid energy. Plasma acetate concentration was determined as well as indicators of protein metabolism: daily and cumulative nitrogen balance, whole body leucine kinetics and rectus muscle and liver fractional protein synthetic rates. No overt toxic effects were observed at any point during the study. As the proportion of short-chain triglycerides in the diet increased from 0 to 50 or 90% of the lipid energy, cumulative nitrogen balance increased 50 or 120%, respectively (P less than 0.05). Whole-body and tissue leucine kinetics (determined during the last 2.5 h of the 7-d study) were unaffected by the lipid composition of the diet. Plasma acetate concentration was not significantly different among groups. These results indicate that incorporation of the short-chain triglyceride, triacetin, in nutritionally balanced total parenteral nutrition formulas improves nitrogen balance with no overt toxic effects. These data indicate that triacetin may have a future role as a parenteral nutrient, and that further studies of its use are warranted.


Neonatal genetically lean and obese pigs respond differently to dietary cholesterol.

The impact of cholesterol exposure in early life on later cholesterol metabolism is not clearly understood. Sixteen newborn genetically lean and obese pigs were fed 0 or 5.0 g cholesterol/kg diet (0 or 0.5%) (liquid diets for 12 d, dry diets thereafter) for 33 d, after which they were all fed 10.0 g cholesterol/kg diet (1.0%) for 23 d. All animals were killed on d 56 and whole-body protein, fat and water were determined on the ground carcass. Dietary cholesterol had no consistent effect on growth rates or body composition. Mean fat content of lean pigs was 15.1% compared with 22.7% for obese pigs; corresponding values were 14.8 and 14.4% for protein and 65.5 and 58.3% for water. Concentrations of plasma total cholesterol, HDL cholesterol and apolipoproteins B and A-1 were increased by 0.5% dietary cholesterol in obese but not in lean piglets, although dietary cholesterol caused HDL and LDL size distribution profiles to shift toward larger-sized components in both strains. Plasma total cholesterol and apolipoprotein B concentrations rose two- to eightfold in all groups after the 1% cholesterol diet was consumed; these changes were accompanied by shifts in LDL and HDL size distribution profiles towards larger-sized components. With 1.0% cholesterol in the diet of all groups, HDL cholesterol concentration increased by approximately 50% in both groups of lean pigs and in obese pigs previously fed cholesterol, but did not increase further in obese pigs previously fed 0.5% cholesterol. The magnitude of the hypercholesterolemic response in lean pigs was blunted by previous exposure to 0.5% dietary cholesterol, but the response was accentuated in obese animals that had been previously exposed to 0.5% dietary cholesterol. These data provide evidence that genetic differences between obese and lean pigs affect their serum lipoprotein responses to high cholesterol intake.


Modification of triacylglycerides and apolipoprotein B in rats fed diets containing whole milk, skim milk and milk proteins.

The objective of this study was to determine the effects of diets containing milk and milk protein fractions on plasma and hepatic lipids, apolipoprotein B mRNA abundance, and plasma apolipoprotein concentrations and lipoprotein composition. Male rats were fed for 6 wk diets that contained (wt/wt) 76% whole milk (WM diet), 55% skim milk (SMFF diet), 22% casein (CAS diet), 22% whey protein isolate (WHY diet) or 55% skim milk-low fat (SMLF diet). The fat concentration in the SMLF diet was 7%. Butter oil (20%) and corn oil (2%) were added to the SMFF, CAS and WHY diets. Plasma and VLDL triacylglycerides in the WM-fed rats were about half of the level in the groups fed the SMFF and SMLF diets, but not significantly different from those of the WHY-fed group. Hepatic triacylglycerides generally were lower in the WM-fed group than in the other groups. Plasma cholesterol concentration did not differ among groups. Plasma apolipoprotein B was significantly lower in the WM-fed group than in rats fed the SMLF, SMFF or WHY diets. However, apolipoprotein B mRNA abundance in the liver and small intestinal mucosa did not differ due to dietary treatment. Thus the lipemic response due to whole milk is not associated with milk protein fractions and may be due to the presence of fat globule membrane in the diet containing whole milk.


Folic acid, 5-methyl-tetrahydrofolate and 5-formyl-tetrahydrofolate exhibit equivalent intestinal absorption, metabolism and in vivo kinetics in rats.

The intestinal absorption and in vivo kinetics of (6S)-[3H]-5-methyl-tetrahydrofolate (5-methyl-H4folate), (6S)-[3H]-5-formyl-H4folate and [3H]folic acid were investigated to determine whether inherent differences exist in the overall bioavailability of these folates in rats. Adult rats (n = 9 per group) were given an intragastric dose of the appropriate folate (50 pmol/100 g body wt) in 50 mmol/L ascorbate (pH 7). Each compound underwent nearly complete absorption within 8 h, and there was no significant difference in the excretion kinetics in relation to the form of folate administered. A biphasic pattern of excretion was observed over the following 8 d. Both urine and feces were important excretory routes. The rapid phase of total isotopic excretion (urinary and fecal) exhibited a half time (t1/2) of 0.11-0.12 d, whereas the t1/2 of the slower phase was 13.4-15.9 d. Isotopic distributions and the pattern of labeled folates in urine and tissues were similar regardless of the form administered. These results indicate that the bioavailability of orally administered folic acid, 5-methyl-H4folate and 5-formyl-H4folate is equivalent in rats under the conditions of this study.


Modification of plasma and hepatic lipids of guinea pigs by feeding high oleic acid pork compared with regular pork.

Meat from such monogastric animals as swine can be modified to substitute monounsaturated fatty acids for saturated fatty acids. Because monounsaturated fatty acids have a beneficial effect on serum lipids as compared with saturated fatty acids, the objective of this study was to assess the effect of modified pork as compared with regular pork on serum and hepatic lipids. Guinea pigs were fed diets containing pork from control diet-fed hogs or from hogs fed a diet containing high oleic acid sunflower oil. The pork provided almost all of the fat in the diets at the level of 4 and 15 g/100 g diet, 10 or 34% energy. The high oleic pork muscle and fat contained 26 and 46% less palmitic and stearic acids (the primary saturated fatty acids), respectively, and 31 and 29% more oleic acid (the primary monounsaturated fatty acid) than the regular pork muscle and fat, respectively. Cholesterol concentration of diets ranged from 0.06 to 0.08% of the diet. Although total serum cholesterol, HDL cholesterol and triglyceride concentrations did not differ due to type of pork, results indicated that serum LDL cholesterol was lower (15%) and hepatic cholesterol was greater (15%) in the high oleic pork, 15% fat group as compared with the control pork 15% fat group. Also, serum LDL cholesterol concentration was higher in the groups fed 15% fat compared with those fed 4% fat. In this study pork modified to have more oleic acid and less saturated fatty acids had a positive effect on tissue lipids when fed to animals.


Efficiency of dietary methionine utilization by young pigs.

An assay was conducted to determine the efficiency of dietary methionine retention for protein accretion in 14-kg pigs. During an 18-d feeding period, pigs were fed a chemically defined amino acid basal diet [147.7 g protein (N x 6.25)/kg, 14.43 MJ metabolizable energy/kg, 4000 mg L-cystine/kg] containing 1150, 1725 or 2300 mg L-methionine/kg. Linear (P less than 0.01) responses in daily gain, daily food intake, daily methionine intake, gain:food ratio and empty body weight gain were obtained as dietary methionine level increased. Representative pigs were killed at the beginning of the assay for whole-body compositional analysis, and the comparative slaughter technique was used to estimate methionine retention. Absolute retention of methionine, protein, water, fat and ash increased linearly (P less than 0.05) with dietary methionine supplementation. Regressing methionine retained on methionine intake resulted in an efficiency of methionine retention above maintenance of 71.7%. The maintenance requirement for methionine was estimated to be 57 mg/d (8 mg/kg body wt 0.75) in the presence of excess dietary cystine. Concentrations of most indispensable amino acids, including methionine, in whole-body protein increased linearly, although glycine and proline concentrations decreased as dietary methionine level increased. This suggests that the ratio of whole-body collagen protein to non-collagen protein decreases as dietary methionine is incremented from deficiency to near adequacy.


Rat serum osteocalcin concentration is determined by food intake and not by inflammation.

Osteocalcin, or bone gla protein, is the major noncollagenous protein in bone. Previous findings of decreased serum osteocalcin concentrations in children with Kwashiorkor led us to analyze the respective influence of nutritional status and inflammation on circulating osteocalcin in growing rats. Food deprivation for 72 h induced a significant 24% decrease in serum osteocalcin. Refeeding produced a rapid rise in serum osteocalcin, which reached control concentrations after 24 h of refeeding. Bone osteocalcin was not affected by these dietary manipulations. The changes in serum osteocalcin were not correlated with serum 1,25-dihydroxycholecalciferol, whereas they could be related to serum 25-hydroxycholecalciferol concentrations. Turpentine injection reduced serum osteocalcin concentration, but pair-feeding showed that this decrease was entirely attributable to spontaneous food restriction and not to inflammation. By contrast, the sensitive nutritional marker, serum transthyretin, was affected by both inflammation and food restriction. These results indicate that serum osteocalcin is closely related to food intake but not to inflammation, suggesting that the dramatic decrease in serum osteocalcin that we previously observed in children with Kwashiorkor is due to malnutrition per se.


Consumption of milk from cows immunized with intestinal bacteria influences age-related changes in immune competence in mice.

Milk was obtained from nonimmunized cows and cows immunized with a mixture of various human gut bacteria. Each milk was administered orally to 2-mo-old C57BL/6 mice at a dose of 150 g.kg-1.d-1 for either 6 or 16 mo. The study group had fewer enteric bacteria and a lower concentration of the serum antibodies against enteric bacteria compared with the control group at 8 and 18 mo of age. Furthermore, the study group at 18 mo old had a higher redirected cytotoxicity of intestinal intraepithelial lymphocytes, a higher proliferative response of mesenteric lymph nodes cells against mitogenic or alloantigenic stimulation and a greater ability of the spleen cells to produce anti-sheep erythrocytes IgG antibody after systemic immunization with sheep erythrocytes. A lower level of autoantibodies was observed at 8 mo and 18 mo of age in the study group compared with the control group. These results suggest that the senescence of the murine immune system may be delayed by consumption of milk from immunized cows.


Bioavailability of vitamin B-6 in pregnant rats.

The cause for reduced plasma pyridoxal phosphate (PLP) concentration during pregnancy is not well understood. In this study, nonpregnant (control) and pregnant rats were gavaged with [3H]pyridoxine for assessment of the intestinal absorption, tissue distribution, metabolic utilization and urinary excretion of the vitamin. In addition, plasma PLP and pyridoxal levels and erythrocyte aspartate aminotransferase (EAST) activity were measured. There was 50% lower plasma PLP concentration and 50% greater EAST activity in the pregnant rats and no difference in the stimulation of EAST with exogenous PLP. There was no difference in the intestinal absorption, hepatic uptake and retention or urinary excretion of the radioisotope. Less than 3% of the oral dose was detected in fetal/uterine tissue of the pregnant rats. Results of this study indicate that reduced plasma PLP concentration during pregnancy is not a result of diminished total vitamin B-6 body pools or fetal sequestration of vitamin B-6. Low plasma PLP during pregnancy may be a result of altered distribution of PLP between the plasma and erythrocytes.


Beta-carotene, retinol and retinyl ester concentrations in serum and selected tissues of ferrets fed beta-carotene.

The concentrations of beta-carotene, retinol and retinyl esters in serum and selected tissues of ferrets fed diets supplemented with beta-carotene (80 micrograms/g wet diet) for 3 wk were determined. The initial concentration of serum beta-carotene was 0.011 +/- 0.006 mumol/L (mean +/- SEM); at the end of the experimental period it was 5.75 +/- 1.60 mumol/L. No significant differences in serum retinol and total retinyl esters were observed between beta-carotene-fed and control ferrets that had been fed an unsupplemented diet. The predominant retinyl esters in serum were retinyl stearate (53%) and retinyl palmitate (35%). Of the tissues analyzed after beta-carotene feeding, the liver contained the highest concentration of beta-carotene (78.8 +/- 18.8 nmol/g). Other tissues that contained beta-carotene in amounts ranging from 17 to 20 nmol/g were adrenals, small intestine, stomach and colon; lesser amounts (6.9 nmol/g) were found in kidneys. Amounts ranging from 1.2 to 2.3 nmol/g were found in muscle, bladder, adipose tissue, lungs and skin; only 0.37 and 0.34 nmol/g were present in brain and eyes, respectively. Thus, like humans, ferrets have the capacity to absorb intact beta-carotene and to store this compound in tissues, especially the liver. However, compared with humans, ferrets have elevated concentrations of retinyl esters in serum, liver and other tissues.


Dietary ascorbic acid depresses plasma and low density lipoprotein lipid peroxidation in genetically scorbutic rats.

The effects of dietary ascorbic acid on plasma lipoprotein and liver lipid peroxide concentrations were examined using ODS od/od rats with a genetic defect in the ability to synthesize ascorbic acid. ODS od/od rats were fed purified diets supplemented with 0 to 300 mg ascorbic acid/kg diet for 21 d. An ascorbic acid-free diet induced body weight loss, depleted ascorbic acid in the plasma and increased thiobarbituric acid-reactive substances in the plasma and liver as compared with rats fed ascorbic acid supplemented diets and with normal ODS +/+ rats fed the ascorbic acid-free diet. Increasing ascorbic acid concentration in the diet inhibited the development of these ascorbic acid deficiency symptoms in a dose-dependent manner. The dietary requirement of ascorbic acid to maintain normal body weight gain and plasma lipid peroxide concentrations was approximately 150 mg ascorbic acid/kg diet. On the other hand, even 300 mg ascorbic acid/kg diet was insufficient to maintain a hepatic concentration of ascorbic acid comparable to that in the liver of ODS +/+ rats. The lipid peroxide concentration in plasma LDL and liver was significantly elevated in ODS od/od rats fed the ascorbic acid-free diet. Supplementing the diet with 300 mg ascorbic acid/kg kept those concentrations within the normal ranges seen in the ODS +/+ rats.


Inhibitory effect of soybean protein vs. casein on apparent absorption of magnesium in rats is due to greater excretion of endogenous magnesium.

Apparent magnesium absorption is depressed in rats fed diets containing soybean protein or enriched with sodium phytate or phosphate in comparison with casein, whereas it is raised in rats fed lactose. However, the possibility that changes in apparent absorption are caused by changes in fecal excretion of endogenous magnesium cannot be excluded. We studied the effects of casein, soybean protein, sodium phytate, lactose and phosphate on apparent and true absorption of magnesium. True magnesium absorption was measured with the use of oral and intraperitoneal administration of tracer doses of 28Mg. Fecal excretion of endogenous magnesium was calculated from apparent and true absorption. True magnesium absorption was not affected by either substitution of soybean protein for casein or by the addition of sodium phytate to a diet containing casein. Endogenous magnesium excretion in feces was significantly increased by soybean protein and sodium phytate. Thus the observed impairment of apparent magnesium absorption in rats fed soybean protein or sodium phytate is due to enhanced fecal excretion of endogenous magnesium. With the other dietary treatments, enhanced fecal excretion of endogenous magnesium was not associated with a discrepancy in the effects on apparent and true magnesium absorption. Dietary lactose vs. dextrose and supplemental phosphate both stimulated fecal excretion of endogenous magnesium, but lactose raised both true and apparent magnesium absorption, and phosphate depressed both true and apparent magnesium absorption.


Annotated guidelines on gifts to physicians from industry. American Medical Association Council on Ethical and Judicial Affairs.

We have responded on an individual basis to many requests for interpretations of grey areas in the opinion on gifts from industry since its release in December 1990, and many physicians and companies asked for a detailed list of these interpretations. While the council agrees with the concerns several individuals have expressed about additional rules, it authorized this revised list of questions and answers which replaces the earlier draft. It also established three main principles for future implementation: 1. The key principles of the guidelines should be carefully observed by physicians, and the AMA will remain active in attempting to secure compliance by its members. The overriding rule is that individual physicians should not accept substantial gifts from industry, even if the gift has an educational or patient benefit. It is important that the profession set clear and enforceable standards in this regard. 2. Professional associations should make their own interpretations of the appropriateness of gifts to them from industry. Under appropriate conditions, associations of physicians may, of course, receive gifts from industry. 3. Neither the council nor its staff will attempt to regulate minor issues or minute details of compliance. For many situations there are no yes or no answers. Some black letter rules are necessary so that conduct that should be changed is changed. In addition, they aid companies which want to comply with the spirit as well as the letter of the guidelines without putting themselves at a competitive disadvantage. The six points of the Opinion cover most situations and compliance to date has been good.(ABSTRACT TRUNCATED AT 250 WORDS)


The state of adult day care in Oklahoma.

Adult daycare is a concept whose time has come, according to the author. And the facts seem to bear him out.


Birth weight-specific fetal deaths and neonatal mortality and the rising cesarean section rate.

To address the question of what effect the rising cesarean section rate has on fetal and neonatal mortality, 33,843 births and 4,132 sections were studied at a community hospital (1968-1987). Crude fetal and neonatal mortality rates declined as the annual section rate rose from 4.7% to 25.7%. However, weight-specific mortality rates bore little relationship to weight-specific section rates. Neonatal mortality rates for infants weighing over 2499 g and without congenital anomalies remained essentially the same (.91, 1.0, 1.02 per 1000 live births) for each of the five-year periods included in the study, despite marked increases in section rates. Among fetal deaths in this weight group, only antepartum deaths declined in each period. This analysis of birth weight-specific mortality indicates that a five-fold rise in the annual cesarean section rate has contributed little, if anything, to reductions in fetal and neonatal mortality during this time.