PubMed Health⌕ Search

Biomedical subjects

Jennifer Jones

Publications and source records attributed to Jennifer Jones.

At least 19 recordsLinked to original sources
PubMed2026

Extracellular Vesicles From Glioblastoma Cells Reflect 2D vs. 3D Culture Adaptation and Resistance to Temozolomide.

Glioblastoma (GBM) is an aggressive brain tumor marked by extensive heterogeneity, resistance to therapy, and dismal prognosis. Extracellular vesicles (EVs) have emerged as key players in GBM biology, mediating intercellular communication and therapy adaptation. However, the exact functions and molecular impact of EVs in GBM remain incompletely understood. In this study, we performed a comparative proteomic analysis of U87MG GBM cells grown in two-dimensional (2D) monolayers and three-dimensional (3D) spheroids following temozolomide (TMZ) treatment, alongside characterization of EVs derived from both culture systems. 3D-spheroids secreted more EVs of smaller size and exhibited a more TMZ-resistant, stem-like proteome under TMZ-induced genotoxic stress. In contrast, 2D cell cultures demonstrated greater proteome remodeling, with EVs enriched in protein families involved in DNA repair, oxidative stress adaptation, and methylation processes. Notably, several methyltransferases were decreased intracellularly but selectively retained in EVs, suggesting active sorting to influence the tumor microenvironment or modulate epigenetic states in recipient cells. EVs also carried adhesion molecules and signaling proteins linked to migration, invasion, and Wnt pathway activation, as well as metabolic enzymes connecting serine metabolism and redox control to TMZ resistance. Mapping EV and cellular proteomes onto The Cancer Genome Atlas (TCGA) dataset identified prognostic protein families associated with either poor or favorable patient outcomes. Our data demonstrate that EV cargo composition mirrors TMZ-induced phenotypic adaptation and reveals molecular mechanisms underlying therapeutic resistance. These EV-associated signatures may serve as clinically actionable biomarkers for patient stratification and offer potential targets to overcome chemoresistance in GBM.

Humans↗
PubMed2006

A high-throughput drug screening system for HIV-1 transcription inhibitors.

Identification of HIV-1 transcription inhibitors was previously performed using infectivity assays. As de novo HIV-1 infection is highly sensitive to even minor compound toxicities, these assays are plagued by extremely high levels of false-positive hits. Hit identification is further complicated because infectivity assays lack target specificity. The authors demonstrate that it is possible to overcome these limitations by establishing a stable, chronically actively HIV-1-infected reporter cell line that exclusively identifies HIV-1 transcription inhibitors. In the reporter cell line, 2 spectrally separated fluorescence proteins serve as simultaneously accessible quantitative markers of HIV-1 expression and drug toxicity. The combined analysis of these markers drastically reduces the level of false-positive hits. As determination of fluorescence intensity in a plate-based format can be performed in a noninvasive manner, repeated measurements of fluorescence levels over several days after compound addition can be used to define the kinetic and dynamic characteristics of inhibitory drug effects. In addition, because of the stable nature of the reporter cell line, the assay requires no cell manipulation during assay preparation or assay analysis, rendering the system extremely cost-effective and reliable.

Animals↗
PubMed2006

Fine organic particulate matter dominates indoor-generated PM2.5 in RIOPA homes.

Residential indoor and outdoor fine particle (PM(2.5)) organic (OC) and elemental carbon (EC) concentrations (48 h) were measured at 173 homes in Houston, TX, Los Angeles County, CA, and Elizabeth, NJ as part of the Relationship of Indoor, Outdoor and Personal Air (RIOPA) study. The adsorption of organic vapors on the quartz fiber sampling filter (a positive artifact) was substantial indoors and out, accounting for 36% and 37% of measured OC at the median indoor (8.2 microg C/m(3)) and outdoor (5.0 microg C/m(3)) OC concentrations, respectively. Uncorrected, adsorption artifacts would lead to substantial overestimation of particulate OC both indoors and outdoors. After artifact correction, the mean particulate organic matter (OM=1.4 OC) concentration indoors (9.8 microg/m(3)) was twice the mean outdoor concentration (4.9 microg/m(3)). The mean EC concentration was 1.1 microg/m(3) both indoors and outdoors. OM accounted for 29%, 30% and 29% of PM(2.5) mass outdoors and 48%, 55% and 61% of indoor PM(2.5) mass in Los Angeles Co., Elizabeth and Houston study homes, respectively. Indirect evidence provided by species mass balance results suggests that PM(2.5) nitrate (not measured) was largely lost during outdoor-to-indoor transport, as reported by Lunden et al. This results in dramatic changes with outdoor-to-indoor transport in the mass and composition of ambient-generated PM(2.5) at California homes. On average, 71% to 76% of indoor OM was emitted or formed indoors, calculated by (1) Random Component Superposition (RCS) model and (2) non-linear fit of OC and air exchange rate data to the mass balance model. Assuming that all particles penetrate indoors (P=1) and there is no particle loss indoors (k=0), a lower bound estimate of 41% of indoor OM was indoor-generated (mean). OM appears to be the predominant species in indoor-generated PM(2.5), based on species mass balance results. Particulate OM emitted or formed indoors is substantial enough to alter the concentration, composition and behavior of indoor PM(2.5). One interesting effect of increased indoor OM concentrations is a shift in the gas-particle partitioning of polycyclic aromatic hydrocarbons (PAHs) from the gas to the particle phase with outdoor-to-indoor transport.

Aerosols↗
PubMed2006

T-cell line for HIV drug screening using EGFP as a quantitative marker of HIV-1 replication.

The rapid increase of viral strains that are resistant to the currently available antiretroviral drugs is a threat to the success of current human immunodeficiency virus type 1 (HIV-1) treatment and emphasizes the importance of developing novel anti-HIV-1 compounds. To improve the current abilities to screen for novel HIV-1 inhibitors, here we introduce a T-cell-based reporter cell line (JLTRG-RS) that expresses both HIV-1 coreceptors, CXCR4 and CCRS, and provides the convenience of using enhanced green fluorescent protein (EGFP) as a direct and quantitative marker. Unlike previous EGFP-based reporter cell lines, JLTRG-RS cells have an unusually high dynamic signal range, sufficient for plate reader detection using a 384-well format. In this format, JLTRG-R5 cell-based infectivity assays have a Z'-factor of 0.78, which defines the assay as extremely robust and clearly amenable to high-throughput screening. The functional similarity of the JLTRG-R5 cell line and peripheral blood mononuclear cells (PBMCs) was demonstrated through the identity of the inhibitory concentrations, 50% (IC50s) for four antiretroviral compounds or neutralizing antibodies. Because EGFP can be directly and continuously quantified in cell culture, the reporter cell line requires no manipulation during assay preparation or analysis. In addition, the EGFP marker allows for data acquisition at an optimal time point by prescreening selected positive control wells using fluorescent microscopy. These characteristics make the system extremely flexible, rapid, and inexpensive. Due to its intrinsic flexibility, the JLTRG-R5 cell-based reporter system provides a powerful tool to greatly facilitate future screening for HIV-1 inhibitors.

Amides↗
PubMed2006

Role of potassium excretion and percent body fat on ethnic differences in plasma aldosterone levels.

OBJECTIVE: To determine whether plasma aldosterone (PA) levels differed between African American and White prehypertensives and if so, could the difference be explained by ethnicity-related variability in urinary K+ and Na+ excretion, body mass index (BMI), and percent body fat. DESIGN: Ethnic comparison SETTING: The University of Maryland College Park and the University of Maryland School of Pharmacy. PARTICIPANTS: 61 (African American, n=28; White, n=33) prehypertensives (systolic blood pressure [SBP] 131 +/- 10 mm Hg, diastolic blood [DBP] 85 +/- 6 mm Hg). INTERVENTION: 6-week dietary stabilization and medication tapering period. MAIN OUTCOME MEASURES: PA levels, Na+ and K+ excretion, blood pressure, and percent body fat and BMI. RESULTS: We saw no differences in SBP (P=.36) and DBP (P=.54) between the two ethnic groups. PA levels were lower in African Americans compared to Whites (62 +/- 7 vs 107 +/- 12 pg/mL, P=.002). 24-hour K+ excretion was lower among African Americans compared to Whites (51 +/- 7 vs 70 +/- 4 mmol/day, P=.002). We saw no difference in percent body fat, BMI, SBP, or DBP between African Americans and Whites. After separately accounting for K+ excretion and Na+ excretion and BMI, plasma aldosterone levels remained significantly different between the two ethnic groups. After adjusting for percent body fat, PA levels were not significantly different between the two ethnic groups (P = .06). CONCLUSIONS: The findings of the current study indicate that PA levels differ between African American and White prehypertensives and this difference may partly be due to ethnic variability in K+ excretion and percent body fat.

Adiposity↗
PubMed2005

Assessment of the amplitude of oscillations associated with high-frequency components of physiological tremor: impact of loading and signal differentiation.

The goal of this study was accurate quantification of the amplitude of high-frequency components of physiological tremor (PT) in units of displacement, velocity, and acceleration. In addition, changes of amplitude with finger loading were compared within specific frequency bands. Index finger tremor was measured for 20 healthy subjects using a high-resolution laser, simultaneously with an accelerometer, under two conditions, unloaded and loaded (70 g). By use of an accurate filtering technique, oscillations within six predetermined frequency bands were isolated. Results showed that overall mean tremor amplitude under the unloaded condition was 0.0973 mm in displacement units, 4.525 mm s(-1) in velocity units, and 301.526 mm s(-2) in acceleration units. Although the mean amplitude of oscillations located within the 16.5-30 Hz band was 0.009 mm and represented only 10% of total tremor amplitude, amplitude of acceleration within the 16.5-30 Hz band was 191 mm s(-2) and represented 60% of total acceleration amplitude. Mean amplitude increased significantly with loading (displacement, t=-2.67, P=0.015; velocity, t=-4.33, P=0.000; acceleration, t=-3.48, P=0.002) but the magnitude of that change was different in each frequency band and its relative importance depended on the level of signal differentiation. Velocity was the only measure that retained sensitivity to changes in amplitude with loading in the low and high-frequency components of PT. In conclusion, this study provides, for the first time, accurate quantification of the amplitude of oscillation of high-frequency components of PT. In addition, it provides clear evidence that the velocity of tremor oscillation is more suitable for detection of the impact of finger loading because it enables detection of amplitude changes in both the low and high-frequency components of PT.

Adolescent↗
PubMed2005

Pattern of nonspecific (or global) DNA methylation in oral carcinogenesis.

BACKGROUND: Although alterations in nonspecific (or global) DNA methylation (GDM) in specific cells are known to be involved in the process of lung carcinogenesis, similar associations have not been evaluated in other smoking-related cancers of the head and neck. METHODS: We evaluated the status of GDM by using monoclonal antibodies specific for 5-methylcytosine (5-mc) in oral squamous cell carcinoma (SCC) specimens of 48 cigarette smokers who had SCC develop and in 93 age-, race-, and sex-matched smokers who did not. RESULTS: Percentages of cells positive for 5-mc immunostaining of DNA of SCC and dysplastic lesions were significantly higher than those of normal oral epithelial cells from cancer subjects and from noncancer subjects. The degree of DNA methylation was unrelated to DNA content. CONCLUSIONS: The pattern of GDM in oral SCCs is different from that of lung SCCs. The differences in nutrient risk factor profiles that are related to GDM and differential activity of DNA methyltranferases between oral and lung SCCs may explain these observations.

5-Methylcytosine↗
PubMed2005

Influence of ambient (outdoor) sources on residential indoor and personal PM2.5 concentrations: analyses of RIOPA data.

The Relationship of Indoor, Outdoor and Personal Air (RIOPA) study was designed to investigate residential indoor, outdoor and personal exposures to several classes of air pollutants, including volatile organic compounds, carbonyls and fine particles (PM2.5). Samples were collected from summer, 1999 to spring, 2001 in Houston (TX), Los Angeles (CA) and Elizabeth (NJ). Indoor, outdoor and personal PM2.5 samples were collected at 212 nonsmoking residences, 162 of which were sampled twice. Some homes were chosen due to close proximity to ambient sources of one or more target analytes, while others were farther from sources. Median indoor, outdoor and personal PM2.5 mass concentrations for these three sites were 14.4, 15.5 and 31.4 microg/m3, respectively. The contributions of ambient (outdoor) and nonambient sources to indoor and personal concentrations were quantified using a single compartment box model with measured air exchange rate and a random component superposition (RCS) statistical model. The median contribution of ambient sources to indoor PM2.5 concentrations using the mass balance approach was estimated to be 56% for all study homes (63%, 52% and 33% for California, New Jersey and Texas study homes, respectively). Reasonable variations in model assumptions alter median ambient contributions by less than 20%. The mean of the distribution of ambient contributions across study homes agreed well for the mass balance and RCS models, but the distribution was somewhat broader when calculated using the mass balance model with measured air exchange rates.

Air Pollutants↗
PubMed2005

Patterns of global DNA and histone methylation appear to be similar in normal, dysplastic and neoplastic oral epithelium of humans.

Although there is growing interest in the possibility that alterations in histone methylation may play a role in carcinogenesis, it has not been explored adequately in humans. Similarly, there are no reports of associations between this and a similar epigenetic event, DNA methylation. Using immunohistochemical staining, we compared the methylation of DNA and histones in histopathologically normal oral epithelium, dysplastic oral lesions, and squamous cell cancers (SCCs) from subjects with squamous cell cancer (n=48) with those of normal oral epithelium from subjects without oral cancer (n=93) who were matched on age and race. Monoclonal antibodies specific for 5 methyl cytosine (5-mc), lysine 4 of histone H_3 (H_3-Lys4), and lysine 9 of histone H_3 (H_3-Lys9) were used in this study. The percentages of cells positive and a weighted average of the immunostaining intensity scores were calculated for each of these tissues, and Spearman correlation analyses were employed to study associations between DNA and histone methylation. Correlations between DNA and histone methylation, H_3-Lys4 and H_3-Lys9 were positive and statistically significant in all tissue types; they were strongest in normal oral epithelium from non-cancer subjects (r=0.63, p < 0.001 and r=0.62, p < 0.001 respectively). Similarly, the positive correlations between H_3-Lys4 and H_3-Lys9 were statistically significant in all tissue types and strongest in normal oral epithelium from non-cancer subjects (r=0.77, p< 0.001). Patterns of DNA and histone methylation are similar in tissues across the spectrum of oral carcinogenesis, and there is a significant positive association between these two epigenetic mechanisms.

Aged↗
PubMed2005

Relationships of Indoor, Outdoor, and Personal Air (RIOPA). Part I. Collection methods and descriptive analyses.

This study on the relationships of indoor, outdoor, and personal air (RIOPA) was undertaken to collect data for use in evaluating the contribution of outdoor sources of air toxics and particulate matter (PM) to personal exposure. The study was not designed to obtain a population-based sample, but rather to provide matched indoor, outdoor, and personal concentrations in homes that varied in their proximity to outdoor pollution sources and had a wide range of air exchange rates (AERs). This design allowed examination of relations among indoor, outdoor, and personal concentrations of air toxics and PM across a wide range of environmental conditions; the resulting data set obtained for a wide range of environmental pollutants and AERs can be used to evaluate exposure models. Approximately 100 households with residents who do not smoke participated in each of three cities in distinct locations expected to have different climates and housing characteristics: Elizabeth, New Jersey; Houston, Texas; and Los Angeles County, California. Questionnaires were administered to characterize homes, neighborhoods, and personal activities that might affect exposures. The concentrations of a suite of volatile organic compounds (VOCs) and carbonyl compounds, as well as the fraction of airborne particulate matter with a mass median aerodynamic diameter < or = 2.5 microm (PM2.5), were measured during continuous 48-hour sessions in which indoor, outdoor, and personal air samples were collected simultaneously. During the same 48-hour period, the AER (exchanges/hr; x hr(-1)) was determined in each home, and carbonyl compounds were measured inside vehicle cabins driven by a subset of the participants. In most of the homes, measurements were made twice, during two different seasons, to obtain a wide distribution of AERs. This report presents in detail the data collection methods, quality control measures, and initial analyses of data distributions and relations among indoor, outdoor, and personal concentrations. The results show that indoor sources dominated personal and indoor air concentrations of many measured VOCs and carbonyl compounds. For several measured species, personal concentrations were higher than either indoor or outdoor concentrations, indicating the presence of some sources closely related to personal activities. For some species there were no significant indoor sources in the majority of the homes; thus indoor concentrations were mainly determined by outdoor concentrations in these homes. The range of distributions of air concentrations for the measured VOCs, formaldehyde and acetaldehyde, PM2.5, and AERs were generally consistent with values reported previously in the literature. Thus associations derived from or models based on this data set that may link the influence of outdoor sources with indoor air concentrations of air toxics and PM2.5 can be relevant to other urban settings. The simultaneous measurements of indoor concentrations, outdoor concentrations, AERs, and room volumes allowed the use of a mass balance model, under the steady-state approximation, to mechanistically examine the relative contributions of indoor and outdoor sources to measured indoor concentrations on a home-by-home basis. Estimated indoor source strengths for VOCs and carbonyl compounds varied widely from home to home, consistent with the indoor-outdoor concentration patterns, as shown in scatter plots. The indoor source estimations agreed with published values for PM2.5 and with the general understanding of sources of VOCs and carbonyl compounds. The source strengths reported here, derived from hundreds of homes, are an important contribution to the literature on exposure to air toxics. For the first time for many compounds, these estimates present a cohesive set of measurements across a range of air toxics in paired indoor, outdoor, and personal samples along with AER and questionnaire results that can be used for future analyses of indoor air quality. The estimation of outdoor contributions to measured indoor concentrations provides insights about the relative importance of indoor and outdoor sources in determining indoor concentrations, the main determinant of personal exposure for most of the measured compounds. In this report simple statistical tests mainly of the pooled data were used to analyze differences by sampling site, emission source type, season, home type, and home age. Paired adult-child personal concentrations within the same home were also compared using the pooled data set. These analyses generated some intriguing results that warrant more in-depth investigation in the future.

Air Movements↗
PubMed2004

CpG-C immunostimulatory oligodeoxyribonucleotide activation of plasmacytoid dendritic cells in rhesus macaques to augment the activation of IFN-gamma-secreting simian immunodeficiency virus-specific T cells.

There are two principle subsets of dendritic cells (DCs); CD11c(+)CD123(-) myeloid DCs (MDCs) and CD11c(-)CD123(+) plasmacytoid DCs (PDCs). DC activation via TNF-TNFRs (e.g., CD40L) and TLRs (e.g., immunostimulatory oligodeoxyribonucleotides (ISS-ODNs)) is crucial for maximal stimulation of innate and adaptive immunity. Macaque DC biology is being studied to improve HIV vaccines using the SIV macaque model. Using lineage (Lin) markers to exclude non-DCs, Lin(-)HLA-DR(+)CD11c(+)CD123(-) MDCs and Lin(-)HLA-DR(+)CD11c(-)CD123(+) PDCs were identified in the blood of uninfected macaques and healthy macaques infected with SIV or simian-human immunodeficiency virus. Overnight culture of DC-enriched Lin-depleted cells increased CD80 and CD86 expression. IL-12 production and CD80/CD86 expression by MDC/PDC mixtures was further enhanced by CD40L and ISS-ODN treatment. A CpG-B ISS-ODN increased CD80/CD86 expression by PDCs, but resulted in little IFN-alpha secretion unless IL-3 was added. In contrast, a CpG-C ISS-ODN and aldrithiol-2-inactivated (AT-2) SIV induced considerable PDC activation and IFN-alpha release without needing exogenous IL-3. The CpG-C ISS-ODN also stimulated IL-12 release (unlike AT-2 SIV) and augmented DC immunostimulatory activity, increasing SIV-specific T cell IFN-gamma production induced by AT-2 SIV-presenting MDC/PDC-enriched mixtures. These data highlight the functional capacities of MDCs and PDCs in naive as well as healthy, infected macaques, revealing a promising CpG-C ISS-ODN-driven DC activation strategy that boosts immune function to augment preventative and therapeutic vaccine efficacy.

2,2'-Dipyridyl↗
PubMed2004

Short-term Flt3L treatment effectively mobilizes functional macaque dendritic cells.

In vivo administration of soluble Flt3L increases dendritic cell (DC) numbers to favor improved DC targeting of vaccine antigens, augmenting vaccine efficiency. In addition to confirming the effectiveness of human Flt3L in macaques, we strove to determine the optimal regimen to elevate numbers of functional DCs. Circulating DCs were identified within lineage(-)human leukocyte antigen-DR(+) cells, which comprised CD11c(-)CD123(+) plasmacytoid DCs (PDCs) and CD123(-) cells including CD11c(+)CD123(-) myeloid DCs as well as CD11c(-)CD123(-) cells. Traditionally, DCs have been monitored 1-2 days after 10- to 14-day treatments with Flt3L (100 microg/kg/day). We demonstrate that although standard treatment increased macaque DC percentages, as little as 5-7 days of treatment was sufficient, if not more effective at mobilizing DCs. Moreover, DC frequency continued to escalate over the ensuing days, peaking at approximately 4 days post 7 days of treatment and ultimately decreasing thereafter. As expected, there was a more pronounced increase in the percentages and actual numbers of CD123(-) cells (CD11c(+) and CD11c(-) subsets) compared with PDCs. Flt3L-mobilized DCs exhibited slightly increased CD80/CD86 expression but typically still that of immature DCs and were resilient to freeze-thawing. Overnight culture activated the cells, up-regulating CD80/CD86 expression as well as interleukin-12 release, typically being boosted by CD40L. This was even more apparent for enriched DC cultures. These data verify that peak mobilization of large numbers of functional macaque DCs occurs a few days, not immediately, after short-term Flt3L dosing. This has important implications for improved DC-targeting vaccine strategies to prevent infection with human immunodeficiency virus and other pathogens.

Adjuvants, Immunologic↗
PubMed2004

Prognostic significance of p27(kip-1) expression in colorectal adenocarcinomas is associated with tumor stage.

PURPOSE: Although the decreased expression of p27(kip-1), a cyclin-dependent kinase inhibitor, has been correlated with advanced tumor stage and short survival of patients with colorectal adenocarcinomas (CRCs), its prognostic value based on the tumor site, tumor stage, and patient ethnicity was not assessed. Therefore, in this study, we investigated whether the prognostic value of p27(kip-1) expression varies with the tumor site, tumor stage and patient ethnicity. EXPERIMENTAL DESIGN: We evaluated 206 (85 African Americans and 121 Caucasians) archival tissue specimens of first primary CRCs for immunohistochemical expression of p27(kip-1), and its prognostic significance was analyzed using univariate Kaplan-Meier and multivariate Cox regression survival methods. RESULTS: Although, similar proportion of CRCs with decreased p27(kip-1) expression was observed in all stages (range, 26-36%), the decreased p27(kip-1) expression has been shown as a marker of poor prognosis only for patients with stage III tumors both in univariate (log-rank test, P = 0.014) and multivariate (hazard ratio = 3.2, 95% confidence interval = 1.3-7.7; P = 0.01) survival analyses. The decreased expression of p27(kip-1) was associated with a high histologic grade (P = 0.016) in stage II CRCs, and with distal tumors (P = 0.001), tumor invasion (P = 0.044), and with local recurrence (P = 0.008) in stage III CRCs. CONCLUSIONS: No prognostic significance was found for p27(kip-1) expression in stages I, II, or IV CRCs, and its prognostic value was not associated with either ethnicity or tumor location. These studies suggest that decreased expression of p27(kip-1) is an indicator of poor prognosis and aids in identifying a subset of patients with aggressive forms of stage III CRCs.

Adenocarcinoma↗
PubMed2004

Primary amyloidosis presenting as small bowel encapsulation.

Amyloidosis is a pathological process which encompasses a spectrum of diseases that result from extracellular deposition of pathological fibrillar proteins. Clinical presentations vary depending on the organs involved. There is no documented case of amyloidosis presenting as small bowel encapsulation. A previously healthy 62-year-old man developed a small bowel obstruction in 1997. At surgery, a peculiar membrane encasing his entire small bowel was discovered. This appeared to have no vascularity and was removed without difficulty, exposing a grossly normal bowel. Histopathology revealed thick bands of collagen overlying the peritoneal surface, which was congo red positive and showed apple green birefringence. The findings were consistent with encapsulating peritonitis due to amyloidosis. There was no history or symptoms of any chronic inflammatory condition and he became symptom-free postoperatively. An abdominal fat pad biopsy failed to demonstrate amyloidosis. Endoscopic duodenal biopsies revealed classical primary amyloidosis. Quantitative immunoglobulins, lactate dehydrogenase, C3, C4 and beta-2 microglobulin were normal. Protein electrophoresis identified monoclonal paraprotein, immunoglobulin G lambda 3.7 g/L. Bone marrow biopsy and aspirate revealed only a mild plasmacytosis (5% to 10%). Echocardiogram and skeletal survey were normal. He had mild proteinuria. Complete blood count, C-reactive protein, calcium, albumin and total protein were normal. No specific therapy was instituted. In January of 1998 the patient remained asymptomatic with no gastrointestinal, cardiovascular or constitutional symptoms. He had developed nephrotic range proteinuria (3.95 g/24 h), microalbuminuria, hypoalbuminemia and a renal biopsy consistent with renal amyloidosis. In 1999 there was an increase in the monoclonal paraprotein (6.2 g/L). The remaining investigations were normal except for an echocardiogram which showed left ventricular hypertrophy but a normal ejection fraction and no diastolic dysfunction. He went on to have high-dose chemotherapy and an autologous stem cell transplant in September, 2000. He has subsequently developed renal insufficiency. To our knowledge this is the first reported case of primary amyloidosis presenting as small bowel obstruction from encapsulating peritonitis.

Amyloidosis↗
PubMed2003

Differential multimerization of Moloney murine leukemia virus integrase purified under nondenaturing conditions.

Retroviral integrases (IN) catalyze the integration of the reverse-transcribed viral DNA into the host genome, an essential process leading to virus replication. For Moloney murine leukemia virus (M-MuLV) IN, the limited solubility of the recombinant protein has restricted the development of biophysical and structural analyses. Herein, recombinant M-MuLV IN proteins, either full length or two nonoverlapping domain constructs, were purified under non-denaturing conditions from solubilized bacterial extracts by Ni(2+)-NTA resins. Additionally, WT IN was further purified by heparin chromatography. All of the purified proteins were shown to be active and stable. WT M-MuLV IN chromatographed with a peak corresponding with a dimer by gel filtration chromatography. In contrast, the single point mutant C209A IN migrated predominantly as a tetramer. For both proteins, fractions in equilibrium between dimers and tetramers were competent to assemble concerted two-end integrations and yielded a unique strand-transfer profile in the presence of a 28-mer U5 oligonucleotide substrate, indicative of a distinct conformation within the synaptic complex. This specific target-site selection was not observed with a shorter 20-mer U5 substrate. These studies provide the foundation for biophysical and structural analysis on M-MuLV IN and the mechanism of retroviral integration.

Amino Acid Sequence↗
PubMed2003

Genetic association of Alzheimer's disease with multiple polymorphisms in alpha-2-macroglobulin.

Alpha-2-Macroglobulin (A2M) is a highly plausible candidate gene for Alzheimer's disease (AD) in a region of chromosome 12 that has numerous independent reports of genetic linkage. We previously reported that a 5 bp deletion in A2M was associated with AD in a subset of the National Institute of Health (NIMH) Genetics Initiative AD family sample. Efforts to replicate this association finding in case - control samples have been largely negative, while those in family samples have been more positive. We hypothesized that variable findings regarding this deletion, along with variable reports of association with V1000I, another polymorphism in the gene, result from linkage disequilibrium in the area as well as ascertainment differences between family-based and case-control studies. Thus, we resequenced the A2M locus to identify novel polymorphisms to test for genetic association with AD. We identified seven novel polymorphisms and tested them in the full NIMH sample of 1439 individuals in 437 families. We found significant genetic association of the 5 bp deletion and two novel polymorphisms with AD. Substantial linkage disequilibrium was detected across the gene as a whole, and haplotype analysis also showed significant association between AD and groups of A2M polymorphisms. Several of these polymorphisms and haplotypes remain significantly associated with AD even after correction for multiple testing. Taken together, these findings, and the positive reports in other family-based studies, continue to support a potential role for A2M or a nearby gene in AD. However, the negative case - control studies suggest that any underlying pathogenic polymorphisms have a modest effect, and may operate primarily among individuals with a family history of AD.

Aged↗
PubMed2003

Prevention of virus transmission to macaque monkeys by a vaginally applied monoclonal antibody to HIV-1 gp120.

A topical microbicide reduces the probability of virus transmission when applied to the vagina or rectum of a person at risk of sexually acquiring HIV-1 infection. An effective microbicide could significantly reduce the global spread of HIV-1, particularly if women were able to use it covertly to protect themselves. A microbicide could target the incoming virus and either permanently inactivate it or reduce its infectivity, or it could block receptors on susceptible cells near the sites of transmission. We describe here how vaginal administration of the broadly neutralizing human monoclonal antibody b12 can protect macaques from simian-human immunodeficiency virus (SHIV) infection through the vagina. Only 3 of 12 animals receiving 5 mg b12 vaginally in either saline or a gel and then challenged vaginally (up to 2 h later) with SHIV-162P4 became infected. In contrast, infection occurred in 12 of 13 animals given various control agents under similar conditions. Lower amounts of b12 were less effective, suggesting that protection was dose dependent. These observations support the concept that viral entry inhibitors can help prevent the sexual transmission of HIV-1 to humans.

Administration, Intravaginal↗