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D M Anderson

Publications and source records attributed to D M Anderson.

At least 55 records · Page 3Linked to original sources

Targeting of Yersinia Yop proteins into the cytosol of HeLa cells: one-step translocation of YopE across bacterial and eukaryotic membranes is dependent on SycE chaperone.

Pathogenic Yersiniae adhere to and kill macrophages by targeting some of their Yop proteins into the eukaryotic cytosol. There is debate about whether YopE targeting proceeds as a direct translocation of polypeptide between cells or in two distinct steps, each requiring specific signals for YopE secretion across the bacterial envelope and for translocation into the eukaryotic cytosol. Here, we used the selective solubilization of the eukaryotic plasma membrane with digitonin to measure Yop targeting during Yersinia infections of HeLa cells. YopE, YopH, YopM and YopN were found in the eukaryotic cytosol but not in the extracellular medium. When bound to SycE chaperone in the Yersinia cytoplasm, YopE residues 1-100 are necessary and sufficient for the targeting of hybrid neomycin phosphotransferase. Electron microscopic analysis failed to detect an extracellular intermediate of YopE targeting, suggesting a one-step translocation mechanism.

Bacterial Outer Membrane Proteins↗

Chemokine receptor (CCR5) expression in human kidneys and in the HIV infected macaque.

BACKGROUND: The chemokine receptor, CCR5, has been identified as an essential co-receptor with CD4, which permits entry of human immunodeficiency virus (HIV) into mammalian cells. This receptor may also mediate leukocyte and parenchymal responses to injury by virtue of its binding to locally released chemokines such as RANTES, MIP-1 alpha and MIP-1 beta during inflammation. The localization of CCR5 in human or primate kidney is unknown. In this study we sought to identify sites of CCR5 synthesis through localization of mRNA coding for this peptide. METHODS: CCR5 cDNA cloned into an expression vector was transcribed into a 1.1 Kb antisense riboprobe that was utilized for in situ hybridization (ISH) and Northern blotting studies. RESULTS: Northern analysis demonstrated positive hybridization for CCR5 mRNA in total RNA isolated from allograft nephrectomy tissue with features of severe transplant rejection as well as in kidney tissue with focal interstitial nephritis. No comparable hybridization signal was achieved with human kidney tissue uninvolved by disease. CCR5 mRNA was not identified in intrinsic renal cell types by ISH in normal human (N = 6), normal macaque kidney (N = 5), in kidneys from macaques with established infection by HIV-2 (N = 9), kidneys from macaques infected with HIV-1 (N = 4), nor in kidneys from SIV-infected macaques (N = 5). CCR5 was identified by ISH in human kidneys with features of interstitial nephritis (N = 3) and in rejected human allograft kidneys (N = 14). The expression of CCR5 was restricted to infiltrating mononuclear leukocytes at sites of chronic tubulointerstitial injury and at sites of vascular and interestitial rejection, respectively. CONCLUSIONS: Understanding the localization of CCR5 as well as other chemokine receptors may help us understand how specificity in leukocyte trafficking is achieved in renal inflammatory processes such as allograft rejection and interstitial nephritis. They provide additional evidence that chemokines may be critical mediators of leukocyte trafficking in renal allograft rejection. These findings may account in part for the difficulty in demonstrating HIV infection of renal cells in human HIV infection, since these cells appear to lack constitutive expression of an essential co-receptor needed for viral entry.

Animals↗

The number of times attended as a factor in the success of a comprehensive hospital-based approach to health promotion.

This study assesses the relationship between a perceived change in a health-related behavior or attitude and the number of times an individual participates in a weekly hospital-based health promotion program. A survey was distributed to participants in April 1995. A univariate probit evaluation shows that the number of times attended and age have positive effects on the likelihood of change. As the number of times an individual attends increases, the more likely the person is to change, possibly revealing a reinforcement mechanism. It is important to learn the most effective way to raise attendance so the reinforcement process reaches more individuals.

Adult↗

Effects of volatile compounds on consumption of alfalfa pellets by sheep.

We examined the effects of six volatile compounds on alfalfa pellet consumption by lambs. In each experiment, 45 lambs were individually fed alfalfa pellets sprayed with a selected compound (camphor, limonene, cis-jasmone, beta-caryophyllene, borneol, or alpha-pinene) at one of five concentrations. Treatment concentrations were multiples (0, .5, 1, 2, and 10) of the concentration of a specific compound (X) that was related to differential herbivory of tarbush by livestock in previous studies. Treatments were applied to alfalfa pellets (.64 kg x lamb(-1) x d(-1), DM basis), and consumption was measured during a 20-min interval each morning for 5 d. Lambs were adapted to handling procedures and the pelleted diet (without treatments) for 10 d. Lambs were maintained and fed (approximately 4.5 to 5% of BW) as one group except during 20-min tests. A negative linear effect of treatment concentration on intake was observed for camphor (P < .02) and alpha-pinene (P < .01), and a quadratic response was detected for borneol (P < .02). The other three compounds had no discernible effect on consumption. Although volatile compounds generally had only minor influences on consumption, the negative influences of alpha-pinene and camphor concentrations on pellet consumption suggest that these monoterpenes may partially explain differential herbivory of individual tarbush plants by livestock.

Administration, Oral↗

A homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function.

Dendritic cells are rare haematopoietic cells that reside in a number of organs and tissues. By capturing, processing and presenting antigens to T cells, dendritic cells are essential for immune surveillance and the regulation of specific immunity. Several members of the tumour necrosis factor receptor (TNFR) superfamily are integral to the regulation of the immune response. These structurally related proteins modulate cellular functions ranging from proliferation and differentiation to inflammation and cell survival or deaths. The functional activity of dendritic cells is greatly increased by signalling through the TNFR family member CD40. Here we report the characterization of RANK (for receptor activator of NF-kappaB), a new member of the TNFR family derived from dendritic cells, and the isolation of a RANK ligand (RANKL) by direct expression screening. RANKL augments the ability of dendritic cells to stimulate naive T-cell proliferation in a mixed lymphocyte reaction, and increases the survival of RANK+ T cells generated with interleukin-4 and transforming growth factor (TGF)-beta. Thus RANK and RANKL seem to be important regulators of interactions between T cells and dendritic cells.

Amino Acid Sequence↗

A mRNA signal for the type III secretion of Yop proteins by Yersinia enterocolitica.

Pathogenic Yersinia species have a specialized secretion system (type III) to target cytotoxic Yop proteins during infection. The signals of YopE and YopN sufficient for the secretion of translational reporter fusions were mapped to the first 15 codons. No common amino acid or peptide sequence could be identified among the secretion signals. Systematic mutagenesis of the secretion signal yielded mutants defective in Yop translation; however, no point mutants could be identified that specifically abolished secretion. Frameshift mutations that completely altered the peptide sequences of these signals also failed to prevent secretion. Thus, the signal that leads to the type III secretion of Yop proteins appears to be encoded in their messenger RNA rather than the peptide sequence.

Amino Acid Sequence↗

Toxin production of Alexandrium minutum (Dinophyceae) from the Bay of Plenty, New Zealand.

Paralytic shellfish toxins of two clonal cultures of Alexandrium minutum isolated during the 1993 toxic shellfish events in the Bay of Plenty, New Zealand, were analyzed using high-performance liquid chromatography. Toxin composition profiles of both cultures showed neosaxitoxin (> 65 mole%) as the principal toxin, with saxitoxin and gonyautoxins (GTX1-4) as minor components. Neither C-toxins (C1-4) nor GTX5-6 were detectable in the two isolates. Bay of Plenty isolates of A. minutum have a unique toxin profile not found in any other isolates of this species that have been characterized. This weakens the hypothesis that A. minutum was recently introduced to New Zealand waters by ballast water or other long-distance transport mechanisms, and argues instead that the species was endemic to the area, but not noticed in the past. The average toxicity of the cultures was 8.8 and 11.0 pg saxitoxin equiv. cell-1 with acetic acid or HCl extraction, respectively. These are at the high end of the range of toxicity reported for A. minutum strains from around the world, and on a cell volume basis are comparable to the most toxic strains of the Alexandrium tamarense group. The toxin profile of A. minutum most closely matches that of mussels and to a lesser degree tuatua harvested from the Bay during the 1993 outbreak, but is quite different from the profile measured in scallops and pipi. Plausible mechanisms for bioconversion of the ingested algal toxins within the latter two shellfish species can be proposed, but it seems more likely that either other strains of A. minutum or other saxitoxin-producing dinoflagellates were ingested by those shellfish. This study established that A. minutum from the Bay of Plenty contains saxitoxins, has a unique toxin composition compared to all other isolates of this species, and was responsible for at least part of the PSP toxicity measured in shellfish during the 1993 outbreak.

Animals↗

Nucleotide sequence of a gene encoding an organophosphorus nerve agent degrading enzyme from Alteromonas haloplanktis.

Organophosphorus acid anhydrolases (OPAA) catalyzing the hydrolysis of a variety of toxic organophosphorus cholinesterase inhibitors offer potential for decontamination of G-type nerve agents and pesticides. The gene (opa) encoding an OPAA was cloned from the chromosomal DNA of Alteromonas haloplanktis ATCC 23821. The nucleotide sequence of the 1.7 -kb DNA fragment contained the opa gene (1.3 kb) and its flanking region. We report structural and functional similarity of OPAAs from A. haloplanktis and Alteromonas sp JD6.5 with the enzyme prolidase that hydrolyzes dipeptides with a prolyl residue in the carboxyl-terminal position. These results corroborate the earlier conclusion that the OPAA is a type of X-Pro dipeptidase, and that X-Pro could be the native substrate for such an enzyme in Alteromonas cells.

Amino Acid Sequence↗

Two independent type III secretion mechanisms for YopE in Yersinia enterocolitica.

Pathogenic Yersinia species escape the infected host's defense mechanisms by targeting cytotoxic Yop proteins into the cytoplasm of macrophages via a type III secretion pathway. Two separate secretion signals contained in YopE were identified, each of which were sufficient but not necessary for the secretion of reporter molecules. One signal is located within the coding sequence of the first 15 amino acids and is sufficient for the secretion of fusion proteins but not required for YopE secretion. The second signal is located downstream at residues 15-100 of YopE and is only recognized by the type III machinery when it is bound to SycE. We propose the existence of two independent mechanisms that allow for the secretion of Yop proteins.

Amino Acid Sequence↗

Progressive multifocal leukoencephalopathy presenting as human immunodeficiency virus type 1 (HIV)-associated dementia.

Progressive multifocal leukoencephalopathy (PML) is a demyelinating disorder of the CNS that usually causes hemiparesis or hemianopsia. Dementia occurs in combination with other neurologic abnormalities. We report a human immunodeficiency virus type 1 (HIV)-infected man whose only manifestation of proven PML was dementia that was clinically indistinguishable from HIV-associated dementia.

AIDS Dementia Complex↗

Embolization of cerebral arteriovenous malformations with silk: histopathologic changes and hemorrhagic complications.

PURPOSE: To evaluate the safety of silk as an embolic agent for preoperative embolization of cerebral arteriovenous malformations (AVMs) by assessing the histopathologic changes and hemorrhagic complications associated with its use. METHODS: Histopathologic specimens, medical records, and radiologic records of 73 patients with AVMs embolized with silk (alone or in combination with other agents) were reviewed retrospectively. Forty-eight histologic specimens obtained at surgery were analyzed for inflammatory responses and compared with the time interval between embolization and surgery. Postembolization angiograms were assessed for vasculitis and CT scans were reviewed for evidence of hemorrhage after embolization. RESULTS: There was no angiographic evidence of vasculitis. Histologic evidence of vasculitis was absent or mild in 92% of cases and histologic evidence of perivascular inflammation was absent or mild in 73% of cases. The frequency of histologic changes associated with vasculitis, perivascular inflammation, and vessel necrosis varied with the time interval between embolization and AVM resection. Intracranial hemorrhage, as a direct complication of silk use, occurred in one patient. Another patient had subarachnoid hemorrhage 24 hours after embolization, caused by rupture of a posteroinferior cerebellar artery aneurysm. Intraventricular high-density material appeared on routine postembolization CT scans in two other patients who had intraventricular AVM extension. This high-density material was thought to be contrast extravasation from intrinsically leaky AVM nidus vessels and not frank hemorrhage. CONCLUSION: Embolization of AVMs with silk does not result in marked inflammation or increased hemorrhagic complications as compared with other agents.

Adolescent↗

Characterization and expression of chitinase and 1,3-beta-glucanase genes in cotton.

We have isolated cDNA clones representing mRNAs encoding chitinase and 1,3-beta-glucanase in cotton (Gossypium hirsutum L.) leaves. The chitinase clones were sequenced and found to encode a 28,806 Da protein with 71% amino acid sequence similarity to the SK2 chitinase from potato (Solanum tuberosum). The 1,3-beta-glucanase clones encoded a 37,645 Da protein with 57.6% identity to a 1,3-beta-glucanase from soybean (Glycine max). Northern blot analyses showed that chitinase mRNA is induced in plants treated with ethaphon or salicylic acid, whereas the levels of 1,3-beta-glucanase mRNA are relatively unaffected. Southern blots of cotton genomic DNA and genomic clones indicated chitinase is encoded by a small gene family of which two members, Chi 2;1 and Chi 2;2, were characterized. These genes share 97% sequence identity in their transcribed regions. The genes were found to have three exons which are 309, 154 and 550 bp long, and two introns 99 and 154 bp in length. The 5'-flanking regions of Chi 2;1 and Chi 2;2 exhibit a large degree of similarity and may contain sequences important for gene response to chemical agents and fungal attack.

Amino Acid Sequence↗

Characterization and expression of metallothionein-like genes in cotton.

We have characterized cotton (Gossypium hirsutum L.) genes encoding type 1 metallothionein-like proteins that are highly expressed in roots. Little or no expression of these genes was detected in other organs and tissues. The deduced amino acid sequences have a high degree of similarity with type 1 metallothionein-like proteins from other plants, including a central hydrophobic domain flanked by conserved cysteine-rich motifs. The type 1 metallothionein-like genes of cotton are encoded by a small gene family. One gene (MT1-A) was analyzed in detail and found to have three exons which are 52, 83 and 397 bp long, and two introns 130 and 1042 bp in length. Three of the type 1 metallothionein-like genes are organized in a tandom array, and the 5'-flanking regions of these genes share a high degree of sequence similarity. Two of the clustered genes (MT1-A and MT1-B) are expressed at about equal levels in roots and use the same transcription start site. A 640 bp promoter fragment from the MT1-A gene was sufficient to direct expression of beta-glucuronidase (GUS) in transformed cotton roots. The expression was highest near the root tip.

Amino Acid Sequence↗

Paralytic shellfish poisoning in southern China.

The rapidly expanding mariculture and commercial region along the southern coast of China has experienced sporadic outbreaks of paralytic shellfish poisoning for nearly 30 years, yet virtually nothing is known of the nature of that toxicity or of the causative organisms. This study presents the first direct comparisons of the high performance liquid chromatography toxin composition profiles of shellfish implicated in paralytic shellfish poisoning outbreaks in Daya Bay with Alexandrium tamarense cultures established from those waters. The three cultures that were analyzed produced an unusually high proportion of the low potency N-sulfocarbamoyl toxins C1 and C2 (nearly 90% of the total), and only trace quantities of the other saxitoxin derivatives. Total toxicity was thus very low with mild acid extraction, ranging between 7.2 and 12.7 fmole cell-1, or 0.7-0.9 pg saxitoxin equiv. cell-1. Following acid hydrolysis using the standard AOAC extraction method, the dominant toxins in the cultures were gonyautoxins 2 and 3 and decarbamoyl gonyautoxins 2 and 3. Total potency increased fourfold to 2.6-3.4 pg saxitoxin equiv. cell-1 following acid hydrolysis. These cultures are thus at the low end of the range of toxicities recorded for members of the A. tamarense species complex. Two scallop samples and one mussel sample collected from Daya Bay during paralytic shellfish poisoning episodes in 1990 and 1991 were also analyzed following the AOAC extraction procedure. The toxin profiles were similar for the three shellfish samples, in that the same suite of toxins were present in each, but the relative proportion of those toxins varied. The dominant toxins were gonyautoxins 2 and 3 and toxins C1-C4. Total toxicity was 336 and 654 micrograms saxitoxin per 100 g meat for the scallop samples, and 723 for the mussels. Toxins C3,4 were present in the shellfish at up to 22 mole%, but were not detected in cultures, even when mild acid was used for extraction. Despite the otherwise similar nature of the culture versus the shellfish toxin signatures, the presence of C3,4 indicates that another strain or species of Alexandrium, or possibly a paralytic shellfish poisoning-producing species of another genus was responsible for the 1990 and 1991 paralytic shellfish poisoning outbreaks in Daya Bay. Since the cultures analyzed were of low intrinsic toxicity, A. tamarense may be more widespread along the south coast of China than is suggested by the sporadic pattern of past paralytic shellfish poisoning outbreaks. Blooms with high cell density are required to generate sufficient toxin to be dangerous. The alarming increase in algal blooms in Chinese waters due to persistent and growing pollution may make these low toxicity populations more problematic in the future.

Animals↗

Developing historical food production and consumption data for 131I dose estimates: the Hanford experience.

This paper describes the methods used to reconstruct the movement of commercial foods in and through the study area of the Hanford Environmental Dose Reconstruction Project. The most dose-relevant radionuclide released from Hanford separations plants was 131I via the atmospheric pathway. As a result of atmospheric deposition of 131I, commercial food supplies may have been contaminated. Because the half-life of 131I is relatively short, foods consumed soon after production, such as milk and produce, presented the highest risk. For that reason, this paper deals primarily with the reconstruction of milk and produce production, marketing, and consumption from 1945-1951, the period with the highest known 131I releases. The reconstructed food production and consumption information was used as input to radiation dose estimates for representative individuals and as default values for real individuals who may not remember where they obtained food or how much they consumed during that period. Specific methods for tracing the movement of commercial milk and produce back from the point of human consumption, through commercial markets, to original production are presented. Results include the characteristics of food consumption exhibited by representative individuals, examples of commercial milk and produce market structures, and a review of commercial milk production and processing practices from 1945-1951.

Air Pollutants, Radioactive↗

Special properties of human milk.

In this review, several nutritional and nonnutritive differences between mothers' milk and formula and their relationship to neonatal gastrointestinal and immune processes are discussed. The dynamic relationship of human milk as evidenced by its changing composition, unique bioactive and immunologic properties, and specialized cellular components is further delineated. The clinical significance and relevance of these findings to the clinician are then presented. Lastly, educational strategies, their effectiveness in promoting breastfeeding, and an approach that might be taken by the clinician to encourage breastfeeding are outlined.

Breast Feeding↗

Expression of interleukin-15 and its receptor by human fetal retinal pigment epithelial cells.

PURPOSE: IL-15 and IL-15 receptor expression was measured in retinal pigment epithelial (RPE) cells to support a possible role of IL-15 in ocular inflammatory and immune responses. METHODS: Reverse transcription-coupled polymerase chain reaction (RT-PCR) and Northern blot analysis of IL-15 mRNA in previously characterized non-transformed and simian virus (SV)-40 transformed human fetal RPE cells were carried out. Biological activities of IL-15 produced by the RPE cells were assayed by co-culture with IL-15 responsive cells. Expression of the IL-15 receptor (IL-15R) alpha, IL-2R beta and gamma chains were examined by RT-PCR. RESULTS: Both non-transformed and SV-40 transformed human fetal RPE cells express IL-15, a T cell growth factor which has similar biological activities to IL-2, and the expression of IL-15 is enhanced by interferon-gamma (IFN-gamma) or tumor necrosis factor-alpha (TNF-alpha) stimulation. In addition, transcripts for all three IL-15 receptor components (IL-15R alpha, IL-2R beta and IL-2R gamma) were detected in these cells. CONCLUSIONS: RPE cells produce IL-15, which may play an important role in ocular immune and inflammatory responses by stimulating infiltrated T cells and RPE cells via paracrine and autocrine loops, respectively.

Blotting, Northern↗