PubMed Health⌕ Search

Biomedical subjects

R L Warren

Publications and source records attributed to R L Warren.

At least 19 recordsLinked to original sources
PubMed2000

Effects of opsonization and gamma interferon on growth of Brucella melitensis 16M in mouse peritoneal macrophages in vitro.

Entry of opsonized pathogens into phagocytes may benefit or, paradoxically, harm the host. Opsonization may trigger antimicrobial mechanisms such as reactive oxygen or nitric oxide (NO) production but may also provide a safe haven for intracellular replication. Brucellae are natural intramacrophage pathogens of rodents, ruminants, dogs, marine mammals, and humans. We evaluated the role of opsonins in Brucella-macrophage interactions by challenging cultured murine peritoneal macrophages with Brucella melitensis 16M treated with complement- and/or antibody-rich serum. Mouse serum rich in antibody against Brucella lipopolysaccharide (LPS) (aLPS) and human complement-rich serum (HCS) each enhanced the macrophage uptake of brucellae. Combinations of suboptimal levels of aLPS (0. 01%) and HCS (2%) synergistically enhanced uptake. The intracellular fate of ingested bacteria was evaluated with an optimal concentration of gentamicin (2 microg/ml) to control extracellular growth but not kill intracellular bacteria. Bacteria opsonized with aLPS and/or HCS grew equally well inside macrophages in the absence of gamma interferon (IFN-gamma). Macrophage activation with IFN-gamma inhibited replication of both opsonized and nonopsonized brucellae but was less effective in inhibiting replication of nonopsonized bacteria. IFN-gamma treatment of macrophages with opsonized or nonopsonized bacteria enhanced NO production, which was blocked by N(G)-monomethyl L-arginine (MMLA), an NO synthesis inhibitor. MMLA also partially blocked IFN-gamma-mediated bacterial growth inhibition. These studies suggest that primary murine macrophages have limited ability to control infection with B. melitensis, even when activated by IFN-gamma in the presence of highly opsonic concentrations of antibody and complement. Additional cellular immune responses, e.g., those mediated by cytotoxic T cells, may play more important roles in the control of murine brucellosis.

Animals↗
PubMed2000

Short report: phylogenetically distinct hepatitis E viruses in Pakistan.

Hepatitis E, which is enterically transmitted, is the most common cause of acute hepatitis in much of Asia. Phylogenetic analysis of several isolates of hepatitis E virus (HEV) from Asia suggests that transmission of this virus is geographically restricted. In Sarghoda, Pakistan, HEV Sar-55 was isolated from a 1987 outbreak. It belongs to the Central-Asian cluster of the Asian sub-genotype. We now report the complete sequence of a second Pakistan HEV from a 1988 outbreak in Abbottabad. The Abbottabad nucleotide sequence was compared with 15 other complete HEV sequences using statistical methods of phylogenetic analysis. The analysis showed that Abbottabad HEV belongs to the South Asia cluster of the Asian sub-genotype. The sequence differences of the 2 Pakistan isolates recovered only one year apart suggest that HEV of 2 distinct origins circulate in Pakistan.

Base Sequence↗
PubMed1999

Protection of mice against brucellosis by vaccination with Brucella melitensis WR201(16MDeltapurEK).

Human brucellosis can be acquired from infected animal tissues by ingestion, inhalation, or contamination of the conjunctiva or traumatized skin by infected animal products. A vaccine to protect humans from occupational exposure or from zoonotic infection in areas where the disease is endemic would reduce an important cause of morbidity worldwide. Vaccines currently used in animals are unsuitable for human use. We tested a live, attenuated, purine-auxotrophic mutant strain of Brucella melitensis, WR201, for its ability to elicit cellular and humoral immune responses and to protect mice against intranasal challenge with B. melitensis 16M. Mice inoculated intraperitoneally with WR201 made serum antibody to lipopolysaccharide and non-O-polysaccharide antigens. Splenocytes from immunized animals released interleukin-2 (IL-2), gamma interferon, and IL-10 when cultured with Brucella antigens. Immunization led to protection from disseminated infection but had only a slight effect on clearance of the challenge inoculum from the lungs. These studies suggest that WR201 should be further investigated as a vaccine to prevent human brucellosis.

Animals↗
PubMed1998

Immunological and molecular characterization of three variant subtype P1.14 strains of Neisseria meningitidis.

Epidemic outbreaks of group B meningococcal disease exhibit a clonal nature consisting of a common serotype-subtype. Subtype-specific monoclonal antibodies (MAbs) directed toward two variable regions (VR1 and VR2) of the class 1 protein of Neisseria meningitidis are used in this classification scheme. A new MAb was developed to classify a nonsubtypeable (NST) strain of N. meningitidis, 7967. This MAb bound to both the NST strain and the prototype subtype P1. 14 strain, S3446, by dot blot analysis. However, a MAb produced to the prototype P1.14 strain did not bind to strain 7967. Sixteen additional strains were further identified as P1.14 with the prototype MAb; of these, 15 strains bound both MAbs. Differences in the characteristics of binding of both antibodies to the three apparently diverse P1.14 strains were studied further by using outer membrane complex proteins, immobilized peptides, and soluble peptides. Deduced amino acid analysis suggested that both MAbs bind to VR2 and that single amino acid changes within VR2 (KM, NM, or KK) might explain the differences in binding characteristics. These results demonstrated that minor variations which exist within subtype variable regions may be clearly identified only by a combination of molecular and immunologic testing. The impact of subtype variation will become more evident as subtype-specific vaccines are developed and tested for efficacy.

Amino Acid Sequence↗
PubMed1997

Characterization of hepatitis E virus (HEV) from Algeria and Chad by partial genome sequence.

The purpose of this study was to analyze partial nucleotide sequences and derived peptide sequences of hepatitis E virus (HEV) from two outbreaks of hepatitis E in Africa (Chad 1983-1984; Algeria 1978-1980). A portion of ORF3 and the major portion of ORF2 were amplified by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). The PCR products were sequenced directly or after cloning into the pCRII vector. Sequences were then compared to the corresponding regions of reported full length HEV sequences. In the ORF2 and ORF3 regions, the homology between the Algerian and the Chad isolates at the nucleic acid level was 92 and 95%, respectively. At the peptide level the homology was 98% in both regions. In these regions, both strains are more related to Asian strains at the nucleic acid level (89 to 95%) and at the amino acid level (95 to 100%) than to the Mexico strain. At the peptide level the differences are less apparent. Both African isolates have amino acid changes in common with some reference strains although the Chad isolate has three unique changes. These African strains of HEV, based on the ORF2 and ORF3 phylogenetic trees, appear to be a distinct phylogenetic group, separate from the Mexican and Asian strains.

Algeria↗
PubMed1997

Predicting patient scores between the functional independence measure and the minimum data set: development and performance of a FIM-MDS "crosswalk".

OBJECTIVE: The functional status of rehabilitation patients is often measured using the Functional Independence Measure (FIM) in acute rehabilitation settings or the Minimum Data Set (MDS) in nursing homes. Because the relationship between the two instruments is unknown, preventing comparison of rehabilitation patients in different types of settings, a translation formula ("crosswalk") between items and subscales from the FIM and the MDS was developed and tested. DESIGN AND OUTCOME MEASURES: Using definitions recommended by an expert panel, MDS items were chosen and rescaled (termed "Pseudo-FIM(E)" items) to correspond to FIM items. The empiric relationships between Pseudo-FIM(E) and FIM scores were then measured using paired FIM-MDS assessments. SETTING AND PATIENTS: 173 rehabilitation patients admitted to six nursing homes. RESULTS: Pseudo-FIM(E) items could be defined for 12 of the 18 FIM items (8 motor and 4 cognitive items). Mean FIM and Pseudo-FIM(E) scores were not significantly different (p > .30) for 5 of the 12 items. Mean scores for the remaining 7 items and for motor and cognitive subscales were similar but statistically significantly different (p < .05). Intraclass correlation coefficients between the FIM and Pseudo-FIM(E) motor and cognitive subscales were both .81. CONCLUSIONS: FIM and MDS items can be used to predict item and subscale scores between the two instruments with reasonable accuracy. This capability will enhance efforts to compare case-mix between acute rehabilitation and nursing home rehabilitation patients, thus making feasible comparisons of the effectiveness (degree of improvement among similar patients) and efficiency (cost of care to obtain a given degree of improvement) of rehabilitation care in different types of settings.

Activities of Daily Living↗
PubMed1997

Hepatitis E virus in Nepal: similarities with the Burmese and Indian variants.

Hepatitis E has been the predominant type of acute hepatitis in Nepal both in adults and children, in sporadic and epidemic forms. We examined six hepatitis E virus (HEV) isolates obtained during an 8-year period, from 1987 to 1995, in the Kathmandu valley of Nepal. Analysis of portions of the putative helicase, polymerase and capsid genes demonstrated close genetic relatedness among themselves (> 96.4% identity) and with the Burmese (> 95.5%) and Indian (> 95.3%) isolates, and less so with the African (> 94.4%) and the Chinese (> 91%) isolates within the Asian genotype. Phylogenetic analysis placed the Nepali isolates in the Burma-India evolutionary branch and showed that the oldest isolate, TK78/87 was more similar to the Burmese isolates whereas the most recent isolates were closer to the Indian ones. Assuming no frameshifts, the Nepali isolates showed high amino acid conservation, but also unique changes when compared to other HEV isolates. Amino acid residue 614 of the capsid protein was identified as a possible marker to distinguish the Burma-Nepal-India from the China-Central Asian Republics subgenotype, and the Mexico genotype.

Adolescent↗
PubMed1997

Interaction of Salmonella typhi strains with cultured human monocyte-derived macrophages.

Human monocyte-derived macrophages (MDM) provided this laboratory with a tool to develop a primary-cell assay for evaluating the relative virulence of newly constructed Salmonella typhi carrier strains. In this study, the interaction with and survival within MDM were compared for delta aroA143-attenuated strains, wild-type virulent strains, and the current oral-vaccine strain, Ty21a.

AIDS Vaccines↗
PubMed1997

Lymphocyte proliferative responses of goats vaccinated with Brucella melitensis 16M or a delta purE201 strain.

The response to a Brucella melitensis purEK deletion mutant, delta purE201 (referred to as strain 201), was compared with the response to its parental strain, 16M, in juvenile goats. Proliferative responses to gamma-irradiated bacteria were detected earlier in strain 201-infected goats. Lymphocytes from strain 16M- or 201-infected goats proliferated in response to one-dimensional polyacrylamide gel electrophoresis-separated proteins of similar mass isolated from strain 16M or Brucella abortus RB51. Data from this study suggest that some antigens stimulating cell-mediated responses are conserved among Brucella species, as 201- and 16M-infected goats recognized similar proteins expressed by RB51 and 16M.

Animals↗