We propose an in vivo method for use with positron emission tomography (PET) that results in a quantitative characterization of neuroleptic binding sites using radiolabeled spiperone. The data are analyzed using a mathematical model that describes transport, nonspecific binding, and specific binding in the brain. The model demonstrates that the receptor quantities Bmax (i.e., the number of binding sites) and KD-<em>1</em> (i.e., the binding affinity) are not separably ascertainable with tracer methodology in human subjects. We have, therefore, introduced a new term, the binding potential, equivalent to the product BmaxKD-<em>1</em>, which reflects the capacity of a given tissue, or region of a tissue, for ligand-binding site interaction. The procedure for obtaining these measurements is illustrated with data from sequential PET scans of baboons after intravenous injection of carrier-added [<em>1</em>8F]spiperone. From these data we estimate the brain tissue nonspecific binding of spiperone to be in the range of 94.2 to 95.3%, and the regional brain spiperone permeability (measured as the permeability-surface area product) to be in the range of 0.025 to 0.036 cm3/(s X <em>ml</em>). The binding potential of the striatum ranged from <em>1</em>7.4 to 2<em>1</em>.6; these in vivo estimates compare favorably to in vitro values in the literature. To our knowledge this represents the first direct evidence that PET can be used to characterize quantitatively, locally and in vivo, drug binding sites in brain. The ability to make such measurements with PET should permit the detailed investigation of diseases thought to result from disorders of receptor function.

BACKGROUND

Cytomegalovirus (CMV) infection is associated with adverse clinical outcomes in immunosuppressed persons, but the incidence and association of CMV reactivation with adverse outcomes in critically ill persons lacking evidence of immunosuppression have not been well defined.

OBJECTIVE

To determine the association of CMV reactivation with intensive care unit (ICU) and hospital length of stay in critically ill immunocompetent persons.

METHODS

We prospectively assessed CMV plasma DNAemia by thrice-weekly real-time polymerase chain reaction (PCR) and clinical outcomes in a cohort of <em>1</em>20 CMV-seropositive, immunocompetent adults admitted to <em>1</em> of 6 ICUs at 2 separate hospitals at a large US tertiary care academic medical center between 2004 and 2006. Clinical measurements were assessed by personnel blinded to CMV PCR results. Risk factors for CMV reactivation and association with hospital and ICU length of stay were assessed by multivariable logistic regression and proportional odds models.

METHODS

Association of CMV reactivation with prolonged hospital length of stay or death.

RESULTS

The primary composite end point of continued hospitalization (n = 35) or death (n = <em>1</em>0) by 30 days occurred in 45 (35%) of the <em>1</em>20 patients. Cytomegalovirus viremia at any level occurred in 33% (39/<em>1</em>20; 95% confidence interval [CI], 24%-4<em>1</em>%) at a median of <em>1</em>2 days (range, 3-57 days) and CMV viremia greater than <em>1</em>000 copies/mL occurred in 20% (24/<em>1</em>20; 95% CI, <em>1</em>3%-28%) at a median of 26 days (range, 9-56 days). By logistic regression, CMV infection at any level (adjusted odds ratio [OR], 4.3; 95% CI, <em>1</em>.6-<em>1</em><em>1</em>.9; P = .005) and at greater than <em>1</em>000 copies/mL (adjusted OR, <em>1</em>3.9; 95% CI, 3.2-60; P < .00<em>1</em>) and the average CMV area under the curve (AUC) in log(<em>1</em>0) copies per milliliter (adjusted OR, 2.<em>1</em>; 95% CI, <em>1</em>.3-3.2; P < .00<em>1</em>) were independently associated with hospitalization or death by 30 days. In multivariable partial proportional odds models, both CMV 7-day moving average (OR, 5.<em>1</em>; 95% CI, 2.9-9.<em>1</em>; P < .00<em>1</em>) and CMV AUC (OR, 3.2; 95% CI, 2.<em>1</em>-4.7; P < .00<em>1</em>) were independently associated with a hospital length of stay of at least <em>1</em>4 days.

CONCLUSIONS

These preliminary findings suggest that reactivation of CMV occurs frequently in critically ill immunocompetent patients and is associated with prolonged hospitalization or death. A controlled trial of CMV prophylaxis in this setting is warranted.

Despite recent advances in the molecular genetics of type 2 diabetes, the majority of susceptibility genes in humans remain to be identified. We therefore conducted a <em>1</em>0-cM genomewide search (40<em>1</em> microsatellite markers) for type 2 diabetes-related traits in 637 members of <em>1</em>43 French pedigrees ascertained through multiple diabetic siblings, to map such genes in the white population. Nonparametric two-point and multipoint linkage analyzes-using the MAPMAKER-SIBS (<em>MLS</em>) and MAXIMUM-BINOMIAL-LIKELIHOOD (MLB) programs for autosomal markers and the ASPEX program for chromosome X markers-were performed with six diabetic phenotypes: diabetes and diabetes or glucose intolerance (GI), as well as with each of the two phenotypes associated with normal body weight (body-mass index<27 kg/m(2)) or early age at diagnosis (<45 years). In a second step, high-resolution genetic mapping ( approximately 2 cM) was performed in regions on chromosomes <em>1</em> and 3 loci showing the strongest linkage to diabetic traits. We found evidence for linkage with diabetes or GI diagnosed at age <45 years in 92 affected sib pairs from 55 families at the D3S<em>1</em>580 locus on chromosome 3q27-qter using MAPMAKER-SIBS (<em>MLS</em> = 4.67, P=.000004), supported by the MLB statistic (MLB-LOD=3.43, P=.00003). We also found suggestive linkage between the lean diabetic status and markers APOA2-D<em>1</em>S484 (<em>MLS</em> = 3. 04, P=.000<em>1</em>8; MLB-LOD=2.99, P=.000<em>1</em>0) on chromosome <em>1</em>q2<em>1</em>-q24. Several other chromosomal regions showed indication of linkage with diabetic traits, including markers on chromosome 2p2<em>1</em>-p<em>1</em>6, <em>1</em>0q26, 20p, and 20q. These results (a) showed evidence for a novel susceptibility locus for type 2 diabetes in French whites on chromosome 3q27-qter and (b) confirmed the previously reported diabetes-susceptibility locus on chromosome <em>1</em>q2<em>1</em>-q24. Saturation on both chromosomes narrowed the regions of interest down to an interval of <7 cM.

We developed two sensitive methods for identifying antimicrobial molecules in leukocytes and other tissues. One method uses a gel overlay technique and was designed to identify antimicrobial polypeptides in samples subjected to polyacrylamide gel electrophoresis. The other, a radial diffusion assay, allows multiple fractions obtained by chromatographic procedures to be tested for antimicrobial activity conveniently. When we used E. coli <em>ML</em>-35p or Salmonella typhimurium <em>1</em>4028S as test organisms in the radial diffusion assay, we routinely detected 5-<em>1</em>0 ng of rabbit defensin NP-<em>1</em> in 5 microliters of sample. With minor modifications, both methods can be applied to other organisms, including Gram-positive bacteria, several Candida species and Cryptococcus neoformans.

BACKGROUND

Biventricular pacing has been proposed to improve symptoms and exercise capacity in patients with advanced heart failure and wide electrocardiographic wave complexes. This study investigated the effect of biventricular pacing on reverse remodeling and the underlying mechanisms.

RESULTS

Twenty-five patients with NYHA class III to IV heart failure and electrocardiographic wave complex duration>><em>1</em>40 ms receiving biventricular pacing therapy were assessed serially up to 3 months after pacing and when pacing was withheld for 4 weeks. Tissue Doppler echocardiography was performed using a 6-basal, 6-mid segmental model to assess the time to peak sustained systolic contraction (T(S)). There was significant improvement of ejection fraction, dP/dt, and myocardial performance index; decrease in mitral regurgitation, left ventricular (LV) end-diastolic (205+/-68 versus <em>1</em>68+/-67 <em>mL</em>, P<0.0<em>1</em>) and end-systolic volume (<em>1</em>62+/-54 versus <em>1</em>22+/-42 <em>mL</em>, P<0.0<em>1</em>); and improved 6-minute hall-walk distance and quality of life score after pacing for 3 months. The mechanisms of benefits were as follows: (<em>1</em>) improved LV synchrony, as evident by homogeneous delay of T(S) to a timing close to the latest (usually the lateral) segment abolishing the intersegmental difference in T(S) and decreasing the standard deviation of T(S) within the left ventricle (37.7+/-<em>1</em>0.9 versus 29.3+/-8.3 ms, P<0.05); (2) improved interventricular synchrony; and (3) shortened isovolumic contraction time (<em>1</em>22+/-57 versus 82+/-36 ms, P<0.05) but increased diastolic filling time. These benefits are pacing dependent, because withholding the pacing resulted in varying speeds in the loss of cardiac improvements.

CONCLUSIONS

Biventricular pacing reverses LV remodeling and improves cardiac function. Improvement of LV mechanical synchrony seems to be the predominant mechanism.

BACKGROUND

In clinical trials, highly active antiretroviral therapy (HAART) reduces plasma HIV-<em>1</em> RNA levels to less than 500 copies/<em>mL</em> in 60% to 90% of patients with HIV-<em>1</em> infection. The performance of such therapy outside of the clinical trial setting is unclear.

OBJECTIVE

To determine factors associated with failure to suppress HIV-<em>1</em> RNA levels and adverse drug reactions in a cohort of patients in whom protease inhibitor-containing therapy was begun in a large urban clinic.

METHODS

Retrospective cohort study.

METHODS

Johns Hopkins HIV Clinic in Baltimore, Maryland.

METHODS

273 protease inhibitor-naive patients began taking a protease inhibitor regimen containing at least one other antiretroviral drug to which the patients had not previously been exposed.

METHODS

Demographic variables, plasma HIV-<em>1</em> RNA levels, CD4+ lymphocyte counts, and adverse drug reactions.

RESULTS

Levels of HIV-<em>1</em> RNA were undetectable in 42% of the cohort at <em>1</em> to 90 days, in 44% at 3 to 7 months, and in 37% at 7 to <em>1</em>4 months. Factors associated with failure to suppress viral load at two or more time points included higher rates of missed clinic appointments, nonwhite ethnicity, age 40 years or younger, injection drug use, lower baseline CD4+ lymphocyte count, and higher baseline viral load. In a multivariate model, only higher rates of missed clinic appointments were independently associated with viral suppression at <em>1</em> year. Ritonavir was associated with adverse drug reactions about twice as frequently as indinavir or nelfinavir, and women experienced significantly more adverse effects than men.

CONCLUSIONS

Unselected patients in whom HAART is started in a clinic setting achieve viral suppression substantially less frequently than do patients in controlled clinical trials. Missed clinic visits were the most important risk factor for failure to suppress HIV-<em>1</em> RNA levels. Studies are needed to identify interventions that maximize the performance of HAART in inner-city clinics.

ESRD incidence is much lower in Europe compared with the United States. This study investigated whether this reflects a difference in the prevalence of earlier stages of chronic kidney disease (CKD) or other mechanisms. CKD prevalence in Norway was estimated from the population-based Health Survey of Nord-Trondelag County (HUNT II), which included 65,<em>1</em>8<em>1</em> adults in <em>1</em>995 through <em>1</em>997 (participation rate 70.4%). Data were analyzed using the same methods as two US National Health and Nutrition Examination Surveys in <em>1</em>988 through <em>1</em>994 (n = <em>1</em>5,488) and <em>1</em>999 through 2000 (n = 4<em>1</em>0<em>1</em>). The primary analysis used gender-specific cutoffs in estimating persistent albuminuria for CKD stages <em>1</em> and 2. ESRD rates and other relevant data were extracted from national registries. Total CKD prevalence in Norway was <em>1</em>0.2% (SE 0.5): CKD stage <em>1</em> (GFR >90 <em>ml</em>/min per <em>1</em>.73 m2 and albuminuria), 2.7% (SE 0.3); stage 2 (GFR 60 to 89 <em>ml</em>/min per <em>1</em>.73 m2 and albuminuria), 3.2% (SE 0.4); stage 3 (GFR 30 to 59 <em>ml</em>/min per <em>1</em>.73 m2), 4.2% (SE 0.<em>1</em>); and stage 4 (GFR <em>1</em>5 to 29 <em>ml</em>/min per <em>1</em>.73 m2), 0.2% (SE 0.0<em>1</em>). This closely approximates reported US CKD prevalence (<em>1</em><em>1</em>.0% in <em>1</em>988 through <em>1</em>994 and <em>1</em><em>1</em>.7% in <em>1</em>999 through 2000). The relative risk for progression from CKD stages 3 or 4 to ESRD in US white patients compared with Norwegian patients was 2.5. This was only modestly modified by adjustment for age, gender, and diabetes. Age and GFR at start of dialysis were similar, hypertension and cardiovascular mortality in the populations were comparable, but US white patients were referred later to a nephrologist and had higher prevalence of obesity and diabetes. In conclusion, CKD prevalence in Norway was similar to that in the United States, suggesting that lower progression to ESRD rather than a smaller pool of individuals at risk accounts for the lower incidence of ESRD in Norway.

BACKGROUND

Previous studies have reported conflicting results concerning the influence of age and gender on the pharmacokinetics and pharmacodynamics of fentanyl, alfentanil, and sufentanil. The aim of this study was to determine the influence of age and gender on the pharmacokinetics and pharmacodynamics of the new short-acting opioid remifentanil.

METHODS

Sixty-five healthy adults (38 men and 27 women) ages 20 to 85 y received remifentanil by constant-rate infusion of <em>1</em> to 8 micrograms.kg-<em>1</em>.min-<em>1</em> for 4 to 20 min. Frequent arterial blood samples were drawn and assayed for remifentanil concentration. The electroencephalogram was used as a measure of drug effect. Population pharmacokinetic and pharmacodynamic modeling was performed using the software package NONMEM. The influence of volunteer covariates were analyzed using a generalized additive model. The performances of the simple (without covariates) and complex (with covariates) models were evaluated prospectively in an additional <em>1</em>5 healthy participants ages 4<em>1</em> to 84 y.

RESULTS

The parameters for the simple three-compartment pharmacokinetic model were V<em>1</em> = 4.98 l, V2 = 9.0<em>1</em> l, V3 = 6.54 l, Cl<em>1</em> = 2.46 l/min, Cl2 = <em>1</em>.69 l/min, and Cl3 = 0.065 l/min. Age and lean body mass were significant covariates. From the ages of 20 to 85 y, V<em>1</em> and Cl<em>1</em> decreased by approximately 25% and 33%, respectively. The parameters for the simple sigmoid Emax pharmacodynamic model were Ke0 = 0.5<em>1</em>6 min-<em>1</em>, E0 = 20 Hz, Emax = 5.62 Hz, EC50 = <em>1</em><em>1</em>.2 ng/ml, and gamma = 2.5<em>1</em>. Age was a significant covariate of EC50 and Ke0, with both decreasing by approximately 50% for the age range studied. The complex pharmacokinetic-pharmacodynamic model performed better than did the simple model when applied prospectively.

CONCLUSIONS

This study identified (<em>1</em>) an effect of age on the pharmacokinetics and pharmacodynamics of remifentanil; (2) an effect of lean body mass on the pharmacokinetic parameters; and (3) no influence of gender on any pharmacokinetic or pharmacodynamic parameter.

The insulin-sensitizing effects of thiazolidinediones are thought to be mediated through peroxisome proliferator-activated receptor-gamma, a nuclear receptor that is highly abundant in adipose tissue. It has been reported that adipocytes secrete a variety of proteins, including tumor necrosis factor-alpha, resistin, plasminogen activator inhibitor-<em>1</em>, and adiponectin. Adiponectin is a fat cell-secreted protein that has been reported to increase fat oxidation and improve insulin sensitivity. Our aim was to study the effects of troglitazone on adiponectin levels in lean, obese, and diabetic subjects. Ten diabetic and <em>1</em>7 nondiabetic subjects (8 lean, BMI <27 kg/m(2) and 9 obese, BMI >27 kg/m(2)) participated in the study. All subjects underwent an 80 mU. m(-2). min(-<em>1</em>) hyperinsulinemic-euglycemic glucose clamp before and after 3 months' treatment with the thiazolidinedione (TZD) troglitazone (600 mg/day). Fasting plasma glucose significantly decreased in the diabetic group after <em>1</em>2 weeks of treatment compared with baseline (9.<em>1</em> +/- 0.9 vs. <em>1</em><em>1</em>.<em>1</em> +/- 0.9 mmol/l, P < 0.005) but was unchanged in the lean and obese subjects. Fasting insulin for the entire group was significantly lower than baseline (P = 0.02) after treatment. At baseline, glucose disposal rate (R(d)) was lower in the diabetic subjects (3.4 +/- 0.5 mg. kg(-<em>1</em>). min(-<em>1</em>)) than in the lean (<em>1</em>2.3 +/- 0.4) or obese subjects (6.7 +/- 0.7) (P < 0.00<em>1</em> for both) and was significantly improved in the diabetic and obese groups (P < 0.05) after treatment, and it remained unchanged in the lean subjects. Baseline adiponectin levels were significantly lower in the diabetic than the lean subjects (9.0 +/- <em>1</em>.7 vs. <em>1</em>6.7 +/- 2.7 micro g/<em>ml</em>, P = 0.03) and rose unifor<em>ml</em>y in all subjects (<em>1</em>2.2 +/- 2.3 vs. 25.7 +/- 2.6 micro g/<em>ml</em>, P < <em>1</em>0(-4)) after treatment, with no significant difference detected among the three groups. During the glucose clamps, adiponectin levels were suppressed below basal levels in all groups (<em>1</em>0.2 +/- 2.3 vs. <em>1</em>2.2 +/- 2.3 micro g/<em>ml</em>, P < 0.0<em>1</em>). Adiponectin levels correlated with R(d) (r = 0.46, P = 0.0<em>1</em>6) and HDL cholesterol levels (r = 0.59, P < 0.00<em>1</em>) and negatively correlated with fasting insulin (r = -0.39, P = 0.042) and plasma triglyceride (r = -0.6<em>1</em>, P < 0.00<em>1</em>). Our findings show that TZD treatment increased adiponectin levels in all subjects, including normal subjects in which no other effects of TZDs are observed. Insulin also appears to suppress adiponectin levels. We have confirmed these results in normal rats. These findings suggest that adiponectin can be regulated by obesity, diabetes, TZDs, and insulin, and it may play a physiologic role in enhancing insulin sensitivity.

A panel of anti-gp<em>1</em>20 human monoclonal antibodies (HuMAbs), CD4-IgG, and sera from people infected with human immunodeficiency virus type <em>1</em> (HIV-<em>1</em>) was tested for neutralization of nine primary HIV-<em>1</em> isolates, one molecularly cloned primary strain (JR-CSF), and two strains (IIIB and MN) adapted for growth in transformed T-cell lines. All the viruses were grown in mitogen-stimulated peripheral blood mononuclear cells and were tested for their ability to infect these cells in the presence and absence of the reagents mentioned above. In general, the primary isolates were relatively resistant to neutralization by the MAbs tested, compared with the T-cell line-adapted strains. However, one HuMAb, IgG<em>1</em>b<em>1</em>2, was able to neutralize most of the primary isolates at concentrations of < or = <em>1</em> microgram/<em>ml</em>. Usually, the inability of a HuMAb to neutralize a primary isolate was not due merely to the absence of the antibody epitope from the virus; the majority of the HuMAbs bound with high affinity to monomeric gp<em>1</em>20 molecules derived from various strains but neutralized the viruses inefficiently. We infer therefore that the mechanism of resistance of primary isolates to most neutralizing antibodies is complex, and we suggest that it involves an inaccessibility of antibody binding sites in the context of the native glycoprotein complex on the virion. Such a mechanism would parallel that which was previously postulated for soluble CD4 resistance. We conclude that studies of HIV-<em>1</em> neutralization that rely on strains adapted to growth in transformed T-cell lines yield the misleading impression that HIV-<em>1</em> is readily neutralized. The more relevant primary HIV-<em>1</em> isolates are relatively resistant to neutralization, although these isolates can be potently neutralized by a subset of human polyclonal or monoclonal antibodies.

BACKGROUND

Rotavirus gastroenteritis causes many deaths in infants in sub-Saharan Africa. Because rotavirus vaccines have proven effective in developed countries but had not been tested in developing countries, we assessed efficacy of a pentavalent rotavirus vaccine against severe disease in Ghana, Kenya, and Mali between April, 2007, and March, 2009.

METHODS

In our multicentre, double-blind, placebo-controlled trial, undertaken in rural areas of Ghana and Kenya and an urban area of Mali, we randomly assigned infants aged 4-<em>1</em>2 weeks without symptoms of gastrointestinal disorders in a <em>1</em>:<em>1</em> ratio to receive three oral doses of pentavalent rotavirus vaccine 2 <em>mL</em> or placebo at around 6 weeks, <em>1</em>0 weeks, and <em>1</em>4 weeks of age. Infants with HIV infection were not excluded. Randomisation was done by computer-generated randomisation sequence in blocks of six. We obtained data for gastrointestinal symptoms from parents on presentation to health-care facilities and clinical data were obtained prospectively by clinicians. The primary endpoint was severe rotavirus gastroenteritis (Vesikari score>>or=<em>1</em><em>1</em>), detected by enzyme immunoassay, arising <em>1</em>4 days or more after the third dose of placebo or vaccine to end of study (March 3<em>1</em>, 2009; around 2<em>1</em> months of age). Analysis was per protocol; infants who received scheduled doses of vaccine or placebo without intervening laboratory-confirmed naturally occurring rotavirus disease earlier than <em>1</em>4 days after the third dose and had complete clinical and laboratory results were included in the analysis. This study is registered with ClinicalTrials.gov, number NCT00362648.

RESULTS

5468 infants were randomly assigned to receive pentavalent rotavirus vaccine (n=2733) or placebo (n=2735). 2357 infants assigned to vaccine and 2348 assigned to placebo were included in the per-protocol analysis. 79 cases of severe rotavirus gastroenteritis were reported in 26<em>1</em>0.6 person-years in the vaccine group, compared with <em>1</em>29 cases in 2585.9 person-years in the placebo group, resulting in a vaccine efficacy against severe rotavirus gastroenteritis of 39.3% (95% CI <em>1</em>9.<em>1</em>-54.7, p=0.0003 for efficacy >0%). Median follow-up in both groups was 527 days starting <em>1</em>4 days after the third dose of vaccine or placebo was given. 42 (<em>1</em>.5%) of 2723 infants assigned to receive vaccine and 45 (<em>1</em>.7%) of 2724 infants assigned to receive placebo had a serious adverse event within <em>1</em>4 days of any dose. The most frequent serious adverse event was gastroenteritis (vaccine <em>1</em>7 [0.6%]; placebo <em>1</em>7 [0.6%]).

CONCLUSIONS

Pentavalent rotavirus vaccine is effective against severe rotavirus gastroenteritis in the first 2 years of life in African countries with high mortality in infants younger than 5 years. We support WHO's recommendation for adoption of rotavirus vaccine into national expanded programmes on immunisation in Africa.

BACKGROUND

PATH (GAVI Alliance grant) and Merck.

BACKGROUND

Probiotic bacteria exhibit a variety of properties, including immunomodulatory activity, which are unique to a particular strain. Thus, not all species will necessarily have the same therapeutic potential in a particular condition. We have preliminary evidence that Bifidobacterium infantis 35624 may have utility in irritable bowel syndrome (IBS).

OBJECTIVE

This study was designed to confirm the efficacy of the probiotic bacteria B. infantis 35624 in a large-scale, multicenter, clinical trial of women with IBS. A second objective of the study was to determine the optimal dosage of probiotic for administration in an encapsulated formulation.

METHODS

After a 2-wk baseline, 362 primary care IBS patients, with any bowel habit subtype, were randomized to either placebo or freeze-dried, encapsulated B. infantis at a dose of <em>1</em> x <em>1</em>0(6), <em>1</em> x <em>1</em>0(8), or <em>1</em> x <em>1</em>0(<em>1</em>0), cfu/<em>mL</em> for 4 wk. IBS symptoms were monitored daily and scored on to a 6-point Likert scale with the primary outcome variable being abdominal pain or discomfort. A composite symptom score, the subject's global assessment of IBS symptom relief, and measures of quality of life (using the IBS-QOL instrument) were also recorded.

RESULTS

B. infantis 35624 at a dose of <em>1</em> x <em>1</em>0(8) cfu was significantly superior to placebo and all other bifidobacterium doses for the primary efficacy variable of abdominal pain as well as the composite score and scores for bloating, bowel dysfunction, incomplete evacuation, straining, and the passage of gas at the end of the 4-wk study. The improvement in global symptom assessment exceeded placebo by more than 20% (p < 0.02). Two other doses of probiotic (<em>1</em> x <em>1</em>0(6) and <em>1</em> x <em>1</em>0(<em>1</em>0)) were not significantly different from placebo; of these, the <em>1</em> x <em>1</em>0(<em>1</em>0) dose was associated with significant formulation problems. No significant adverse events were recorded.

CONCLUSIONS

B. infantis 35624 is a probiotic that specifically relieves many of the symptoms of IBS. At a dosage level of <em>1</em> x <em>1</em>0(8) cfu, it can be delivered by a capsule making it stable, convenient to administer, and amenable to widespread use. The lack of benefits observed with the other dosage levels of the probiotic highlight the need for clinical data in the final dosage form and dose of probiotic before these products should be used in practice.

BACKGROUND

HLA-B57, as well as cytotoxic T-lymphocyte (CTL) responses restricted by this allele, have been strongly associated with long-term non-progressive chronic HIV-<em>1</em> infection. However, their impact on viral replication during acute HIV-<em>1</em> infection is not known.

METHODS

Clinical and immunological parameters during acute and early HIV-<em>1</em> infection in individuals expressing HLA-B57 were assessed. HIV-<em>1</em>-specific T-cell responses were determined by peptide-specific interferon-gamma production measured using Elispot assay and flow-based intracellular cytokine quantification.

RESULTS

Individuals expressing HLA-B57 presented significantly less frequently with symptomatic acute HIV-<em>1</em> infection (4/<em>1</em><em>1</em>6, 3.4%) than expected from the frequency of chronically infected individuals expressing this allele (43/446, 9.6%; P < 0.05). During acute infection, virus-specific CD8 T-cell responses were dominated by HLA-B57-restricted responses, with significantly broader (P < 0.02) and stronger (P < 0.03) responses restricted by HLA-B57 than restricted by all other co-expressed HLA class I alleles combined. Six out of nine individuals expressing HLA-B57 controlled HIV-<em>1</em> viremia in the absence of therapy at levels < 5000 copies/ml (median, 5<em>1</em>5 copies/ml) during up to 29 months following acute infection.

CONCLUSIONS

These data demonstrate that host genetic factors can influence the clinical manifestations of acute HIV-<em>1</em> infection and provide a functional link between HLA-B57 and viral immune control.

BACKGROUND

Endothelium-dependent vasodilation is abnormal in experimental models of diabetes mellitus. We postulated that abnormalities of endothelial function are also present in patients with insulin-dependent diabetes mellitus and may contribute to the pathogenesis of vascular disease in these individuals.

RESULTS

Vascular reactivity was measured in the forearm resistance vessels of <em>1</em>5 patients with insulin-dependent diabetes mellitus and <em>1</em>6 age-matched normal subjects. No patient had hypertension or dyslipidemia. Each subject was pretreated with aspirin to inhibit endogenous production of prostanoids. Methacholine chloride (0.3 to <em>1</em>0 micrograms/min) was administered via the brachial artery to assess endothelium-dependent vasodilation. Sodium nitroprusside (0.3 to <em>1</em>0 micrograms/min) and verapamil (<em>1</em>0 to 300 micrograms/min) were infused intra-arterially to assess endothelium-independent vasodilation; phenylephrine (0.3 to 3 micrograms/min) was administered to examine vasoconstrictor responsiveness. Forearm blood flow was determined by venous occlusion plethysmography, and dose-response curves were generated for each drug. Basal forearm blood flow in diabetic and normal subjects was comparable (2.6 +/- 0.2 versus 2.<em>1</em> +/- 0.3 <em>mL</em> x <em>1</em>00 <em>mL</em>-<em>1</em> x min-<em>1</em>, respectively; P = NS). The forearm vasodilative response to methacholine was less in diabetic than in normal subjects. At the highest dose of methacholine, the forearm blood flow increased 9.5 +/- <em>1</em>.<em>1</em> <em>mL</em> x <em>1</em>00 <em>mL</em>-<em>1</em> x min-<em>1</em> in diabetic subjects and <em>1</em>5.3 +/- <em>1</em>.4 <em>mL</em>.<em>1</em>00 <em>mL</em>-<em>1</em> x min-<em>1</em> in normal subjects (P < .0<em>1</em>). The forearm blood flow responses to nitroprusside and verapamil and the forearm vasoconstrictor responses to phenylephrine were similar in diabetic and healthy subjects. In diabetic subjects, endothelium-dependent vasodilation correlated inversely with serum insulin concentration but not with glucose concentration, glycosylated hemoglobin, or duration of diabetes.

CONCLUSIONS

Endothelium-dependent vasodilation is abnormal in forearm resistance vessels of patients with insulin-dependent diabetes mellitus. This abnormality may be relevant to the high prevalence of vascular disease that occurs in these individuals.

Glucagon-like peptide <em>1</em> (GLP-<em>1</em>) has been shown to inhibit gastric emptying of liquid meals in type 2 diabetic patients. It was the aim of the present study to compare the action of physiological and pharmacological doses of intravenous GLP-<em>1</em>-(7-36) amide and GLP-<em>1</em>-(7-37) on gastric emptying in normal volunteers. Nine healthy subjects participated (26 +/- 3 yr; body mass index 22.9 +/- <em>1</em>.6 kg/m2; hemoglobin A<em>1</em>C 5.0 +/- 0.2%) in five experiments on separate occasions after an overnight fast. A nasogastric tube was positioned for the determination of gastric volume by use of a dye-dilution technique (phenol red). GLP-<em>1</em>-(7-36) amide (0.4, 0.8, or <em>1</em>.2 pmol.kg-<em>1</em>.min-<em>1</em>), GLP-<em>1</em>-(7-37) (<em>1</em>.2 pmol.kg-<em>1</em>.min-<em>1</em>), or placebo was infused intravenously from -30 to 240 min. A liquid meal (50 g sucrose, 8% amino acids, 440 <em>ml</em>, 327 kcal) was administered at 0 min. Glucose, insulin, and C-peptide were measured over 240 min. Gastric emptying was dose dependently slowed by GLP-<em>1</em>-(7-36) amide (P < 0.000<em>1</em>). Effects of GLP-<em>1</em>-(7-37) at <em>1</em>.2 pmol.kg-<em>1</em>.min-<em>1</em> were virtually identical. GLP.<em>1</em> dose dependently stimulated fasting insulin secretion (-30 to 0 min) and slightly reduced glucose concentrations. After the meal (0-240 min), integrated incremental glucose (P < 0.000<em>1</em>) and insulin responses (P = 0.0<em>1</em>) were reduced (dose dependently) rather than enhanced. In conclusion, <em>1</em>) GLP-<em>1</em>-(7-36) amide or -(7-37) inhibits gastric emptying also in normal subjects, 2) physiological doses (0.4 pmol.kg-<em>1</em>.min-<em>1</em>) still have a significant effect, 3) despite the known insulinotropic actions of GLP-<em>1</em>-(7-36) amide and -(7-37), the net effect of administering GLP-<em>1</em> with a meal is no change or a reduction in meal-related insulin responses. These findings suggest a primarily inhibitory function for GLP-<em>1</em> (ileal brake mechanisms).

To determine the incidence of Candida bloodstream infections (BSI) and antifungal drug resistance, population-based active laboratory surveillance was conducted from October <em>1</em>998 through September 2000 in two areas of the United States (Baltimore, Md., and the state of Connecticut; combined population, 4.7 million). A total of <em>1</em>,<em>1</em>43 cases were detected, for an average adjusted annual incidence of <em>1</em>0 per <em>1</em>00,000 population or <em>1</em>.5 per <em>1</em>0,000 hospital days. In 28% of patients, Candida BSI developed prior to or on the day of admission; only 36% of patients were in an intensive care unit at the time of diagnosis. No fewer than 78% of patients had a central catheter in place at the time of diagnosis, and 50% had undergone surgery within the previous 3 months. Candida albicans comprised 45% of the isolates, followed by C. glabrata (24%), C. parapsilosis (<em>1</em>3%), and C. tropicalis (<em>1</em>2%). Only <em>1</em>.2% of C. albicans isolates were resistant to fluconazole (MIC,>> or = 64 microg/<em>ml</em>), compared to 7% of C. glabrata isolates and 6% of C. tropicalis isolates. Only 0.9% of C. albicans isolates were resistant to itraconazole (MIC,>> or = <em>1</em> micro g/<em>ml</em>), compared to <em>1</em>9.5% of C. glabrata isolates and 6% of C. tropicalis isolates. Only 4.3% of C. albicans isolates were resistant to flucytosine (MIC,>> or = 32 microg/<em>ml</em>), compared to < <em>1</em>% of C. parapsilosis and C. tropicalis isolates and no C. glabrata isolates. As determined by E-test, the MICs of amphotericin B were>> or = 0.38 microg/<em>ml</em> for <em>1</em>0% of Candida isolates,>> or =<em>1</em> microg/<em>ml</em> for <em>1</em>.7% of isolates, and>> or = 2 microg/<em>ml</em> for 0.4% of isolates. Our findings highlight changes in the epidemiology of Candida BSI in the <em>1</em>990s and provide a basis upon which to conduct further studies of selected high-risk subpopulations.

The latrunculins are architecturally novel marine compounds isolated from the Red Sea sponge Latrunculia magnifica. In vivo, they alter cell shape, disrupt microfilament organization, and inhibit the microfilament-mediated processes of fertilization and early development. In vitro, latrunculin A was recently found to affect the polymerization of pure actin in a manner consistent with the formation of a <em>1</em>:<em>1</em> molar complex with G-actin. These in vitro effects as well as previous indications that the latrunculins are more potent than the cytochalasins suggest differences in the in vivo mode of action of the two classes of drugs. To elucidate these differences we have compared the short- and long-term effects of latrunculins on cell shape and actin organization to those of cytochalasin D. Exposure of hamster fibroblast NIL8 cells for <em>1</em>-3 hr to latrunculin A, latrunculin B, and cytochalasin D causes concentration-dependent changes in cell shape and actin organization. However, the latrunculin-induced changes were strikingly different from those induced by cytochalasin D. Furthermore, while initial effects were manifest with both latrunculin A and cytochalasin D already at concentrations of about 0.03 microgram/<em>ml</em>, latrunculin A caused complete rounding up of all cells at 0.2 microgram/<em>ml</em>, whereas with cytochalasin D maximum contraction was reached at concentrations <em>1</em>0-20 times higher. The short-term effects of latrunculin B were similar to those of latrunculin A although latrunculin B was slightly less potent. All three drugs inhibited cytokinesis in synchronized cells, but their long-term effects were markedly different. NIL8 cells treated with latrunculin A maintained their altered state for extended periods. In contrast, the effects of cytochalasin D progressed with time in culture, and the latrunculin B-induced changes were transient in the continued presence of the drug. These transient effects were found to be due to a gradual inactivation of latrunculin B by serum and were used to compare recovery patterns of cell shape and actin organization in two different cell lines. This comparison showed that the transient effects of latrunculin B were fully reversible for the NIL8 cells and not for the mouse neuroblastoma N<em>1</em>E-<em>1</em><em>1</em>5 cells.

Recently, we described a simple procedure, Drinking in the Dark (DID), in which C57BL/6J mice self-administer ethanol to a blood ethanol concentration (BEC) above <em>1</em> mg/<em>ml</em>. The test consists of replacing the water with 20% ethanol in the home cage for 4 h early during the dark phase of the light/dark cycle. Three experiments were conducted to explore this high ethanol drinking model further. In experiment <em>1</em>, a microanalysis of C57BL/6J behavior showed that the pattern of ethanol drinking was different from routine water intake. In experiment 2, drinking impaired performance of C57BL/6J on the accelerating rotarod and balance beam. In experiment 3, <em>1</em>2 inbred strains were screened to estimate genetic influences on DID and correlations with other traits. Large, reliable differences in intake and BEC were detected among the strains, with C57BL/6J showing the highest values. Strain means were positively correlated with intake and BEC in the standard (24 h) and a limited (4 h) two-bottle ethanol vs. water test, but BECs reached higher levels for DID. Strain mean correlations with other traits in the Mouse Phenome Project database supported previously reported genetic relationships of high ethanol drinking with low chronic ethanol withdrawal severity and low ethanol-conditioned taste aversion. We extend these findings by showing that the correlation estimates remain relatively unchanged even after correcting for phylogenetic relatedness among the strains, thus relaxing the assumption that the strain means are statistically independent. We discuss applications of the model for finding genes that predispose pharmacologically significant drinking in mice.

More than 80% of the world's HIV-infected adults live in sub-Saharan Africa, where heterosexual transmission is the predominant mode of spread. The virologic and immunologic correlates of female-to-male (FTM) and male-to-female (MTF) transmission are not well understood. A total of <em>1</em>022 heterosexual couples with discordant HIV-<em>1</em> serology results (one partner HIV infected, the other HIV uninfected) were enrolled in a prospective study in Lusaka, Zambia and monitored at 3-month intervals. A nested case-control design was used to compare <em>1</em>09 transmitters and 208 nontransmitting controls with respect to plasma HIV-<em>1</em> RNA (viral load, VL), virus isolation, and CD4(+) cell levels. Median plasma VL was significantly higher in transmitters than nontransmitters (<em>1</em>23,507 vs. 5<em>1</em>,3<em>1</em>0 copies/<em>ml</em>, p < 0.00<em>1</em>). In stratified multivariate Cox regression analyses, the risk ratio (RR) for FTM transmission was 7.6 (95% CI: 2.3, 25.5) for VL>> or = <em>1</em>00,000 copies/<em>ml</em> and 4.<em>1</em> (95% CI: <em>1</em>.2, <em>1</em>4.<em>1</em>) for VL between <em>1</em>0,000 and <em>1</em>00,000 copies/<em>ml</em> compared with the reference group of (<em>1</em>0,000 copies/<em>ml</em>. Corresponding RRs for MTF transmission were 2.<em>1</em> and <em>1</em>.2, respectively, with 95% CI both bounding <em>1</em>. Only 3 of 4<em>1</em> (7%) female transmitters had VL < <em>1</em>0,000 copies/<em>ml</em> compared with 32 of 93 (34%) of female nontransmitters (p < 0.00<em>1</em>). The transmission rate within couples was 7.7/<em>1</em>00 person-years and did not differ from FTM (6<em>1</em>/862 person-years) and MTF (8<em>1</em>/978 person-years) transmission. We conclude that the association between increasing plasma viral load was strong for female to male transmission, but was only weakly predictive of male to female transmission in Zambian heterosexual couples. FTM and MTF transmission rates were similar. These data suggest gender-specific differences in the biology of heterosexual transmission.

In view of the fact that insulin resistance is associated with atherogenesis and that troglitazone, an insulin sensitizer, has anti-inflammatory effects, which may be potentially antiatherogenic in the long term, we have now investigated whether insulin has potential anti-inflammatory effects. We infused 2.0 to 2.5 IU/h in 5% dextrose (<em>1</em>00 <em>mL</em>/h) iv into <em>1</em>0 obese subjects for 4 h followed by 5% dextrose alone for 2 h. The rate of insulin infusion was varied to maintain glucose concentrations as close to the baseline as possible. Blood samples were obtained before and at 2, 4, and 6 h. Subjects were also infused with 5% dextrose without insulin and with saline on separate occasions. Intranuclear nuclear factor kappaB (NFkappaB) in mononuclear cells fell at 2 and further at 4 h, reverting toward the baseline at 6 h (P < 0.05). IkappaB increased significantly at 2 h, increasing further at 4 h and remaining elevated at 6 h (P < 0.00<em>1</em>). Reactive oxygen species (ROS) generation by mononuclear cells fell significantly at 2 h and fell further at 4 h; it partially reverted to baseline at 6 h (P < 0.005). p47(phox) subunit, the key protein of nicotinamide adenine dinucleotide phosphate oxidase also fell at 2 h and 4 h, reverting toward the baseline at 6 h (P < 0.05). In addition, soluble intercellular adhesion molecule-<em>1</em> (sICAM-<em>1</em>), monocyte chemoattractant protein-<em>1</em> (MCP-<em>1</em>), and plasminogen activator inhibitor-<em>1</em> (PAI-<em>1</em>) fell significantly following insulin infusion. Glucose or saline infusions without insulin caused no alteration in NFkappaB, IkappaB, ROS generation, p47(phox) subunit, sICAM-<em>1</em>, MCP-<em>1</em>, or PAI-<em>1</em>. We conclude that insulin has a potent acute anti-inflammatory effect including a reduction in intranuclear NFkappaB, an increase in IkappaB, and decreases in ROS generation, p47(phox) subunit, plasma soluble intercellular adhesion molecule-<em>1</em> (sICAM-<em>1</em>), monocyte chemoattractant protein-<em>1</em> (MCP-<em>1</em>), and plasminogen activator inhibitor-<em>1</em> (PAI-<em>1</em>. This acute anti-inflammatory effect, if demonstrated in the long term, may have implications for atherosclerosis and its complications.

A one-step simple synthesis of silver colloid nanoparticles with controllable sizes is presented. In this synthesis, reduction of [Ag(NH(3))(2)](+) complex cation by four saccharides was performed. Four saccharides were used: two monosaccharides (glucose and galactose) and two disaccharides (maltose and lactose). The syntheses performed at various ammonia concentrations (0.005-0.20 mol L(-<em>1</em>)) and pH conditions (<em>1</em><em>1</em>.5-<em>1</em>3.0) produced a wide range of particle sizes (25-450 nm) with narrow size distributions, especially at the lowest ammonia concentrations. The average size, size distribution, morphology, and structure of particles were determined by dynamic light scattering (DLS), transmission electron microscopy (TEM), and UV/Visible absorption spectrophotometry. The influence of the saccharide structure (monosacharides versus disaccharides) on the size of silver particles is briefly discussed. The reduction of [Ag(NH(3))(2)](+) by maltose produced silver particles with a narrow size distribution with an average size of 25 nm, which showed high antimicrobial and bactericidal activity against Gram-positive and Gram-negative bacteria, including highly multiresistant strains such as methicillin-resistant Staphylococcus aureus. Antibacterial activity of silver nanoparticles was found to be dependent on the size of silver particles. A very low concentration of silver (as low as <em>1</em>.69 mug/<em>mL</em> Ag) gave antibacterial performance.

OBJECTIVE

To evaluate whether two commonly used newer platinum-based regimens offer any advantage over vinorelbine-cisplatin (reference regimen) in response rate for patients with advanced non-small-cell lung cancer (NSCLC).

METHODS

Chemotherapy-naive patients were randomized to receive gemcitabine <em>1</em>,250 mg/m(2) days <em>1</em> and 8 plus cisplatin 75 mg/m(2) day 2 every 2<em>1</em> days (GC arm), or paclitaxel 225 mg/m(2) (3-hour infusion) then carboplatin (area under the concentration-time curve of 6 mg/<em>mL</em> x min), both on day <em>1</em> every 2<em>1</em> days (PCb arm), or vinorelbine 25 mg/m(2)/wk for <em>1</em>2 weeks then every other week plus cisplatin <em>1</em>00 mg/m(2) day <em>1</em> every 28 days (VC arm).

RESULTS

Six hundred twelve patients were randomized to treatment (205 GC, 204 PCb, and 203 VC). Overall response rates for the GC (30%) and PCb (32%) arms were not significantly different from that of the VC arm (30%). There were no differences in overall survival, time to disease progression, or time to treatment failure. Median survival for the GC, PCb, and VC groups was 9.8, 9.9, and 9.5 months, respectively. Neutropenia was significantly higher on the VC arm (GC <em>1</em>7% or PCb 35% v VC 43% of cycles, P <.00<em>1</em>), as was thrombocytopenia on the GC arm (GC <em>1</em>6% v VC 0.<em>1</em>% of cycles, P <.00<em>1</em>). Alopecia and peripheral neurotoxicity were most common on the PCb arm, as was nausea/vomiting on the VC arm (P <.05).

CONCLUSIONS

Efficacy end points were not significantly different between experimental and reference arms, although toxicities showed differences. These findings suggest that chemotherapy in NSCLC has reached a therapeutic plateau.

BACKGROUND

Benign prostatic hyperplasia is a progressive, androgen-dependent disease resulting in enlargement of the prostate gland and urinary obstruction. Preventing the conversion of testosterone to its tissue-active form, dihydrotestosterone, by inhibiting the enzyme 5 alpha-reductase could decrease the action of androgens in their target tissues; in the prostate the result might be a decrease in prostatic hyperplasia and therefore in symptoms of urinary obstruction.

METHODS

In a double-blind study, we evaluated the effect of two doses of finasteride (<em>1</em> mg and 5 mg) and placebo, each given once daily for <em>1</em>2 months, in 895 men with prostatic hyperplasia. Urinary symptoms, urinary flow, prostatic volume, and serum concentrations of dihydrotestosterone and prostate-specific antigen were determined periodically during the treatment period.

RESULTS

As compared with the men in the placebo group, the men treated with 5 mg of finasteride per day had a significant decrease in total urinary-symptom scores (P less than 0.00<em>1</em>), an increase of <em>1</em>.6 ml per second (22 percent, P less than 0.00<em>1</em>) in the maximal urinary-flow rate, and a <em>1</em>9 percent decrease in prostatic volume (P less than 0.00<em>1</em>). The men treated with <em>1</em> mg of finasteride per day did not have a significant decrease in total urinary-symptom scores, but had an increase of <em>1</em>.4 ml per second (23 percent) in the maximal urinary-flow rate, and an <em>1</em>8 percent decrease in prostatic volume. The men given placebo had no changes in total urinary-symptom scores, an increase of 0.2 ml per second (8 percent) in the maximal urinary-flow rate, and a 3 percent decrease in prostatic volume. The frequency of adverse effects in the three groups was similar, except for a higher incidence of decreased libido, impotence, and ejaculatory disorders in the finasteride-treated groups.

CONCLUSIONS

The treatment of benign prostatic hyperplasia with 5 mg of finasteride per day results in a significant decrease in symptoms of obstruction, an increase in urinary flow, and a decrease in prostatic volume, but at a slightly increased risk of sexual dysfunction.

Mesenchymal stem cells (MSCs) residing in bone marrow (BM) are the progenitors for osteoblasts and for several other cell types. In humans, the age-related decrease in bone mass could reflect decreased osteoblasts secondary to an age-related loss of osteoprogenitors. To test this hypothesis, BM cells were isolated from vertebral bodies of thoracic and lumbar spine (T<em>1</em>-L5) from 4<em>1</em> donors (<em>1</em>6 women and 25 men) of various ages (3-70 years old) after death from traumatic injury. Primary cultures were grown in alpha modified essential medium with fetal bovine serum for <em>1</em>3 days until adherent cells formed colonies (CFU-Fs). Colonies that stained positive for alkaline phosphatase activity (CFU-F/ALP+) were considered to have osteogenic potential. BM nucleated cells were plated (0.5, <em>1</em>, 2.5, 5, or <em>1</em>0 x <em>1</em>06 cells/<em>1</em>0-cm dish) and grown in dexamethasone (Dex), which promotes osteoblastic differentiation. The optimal plating efficiency using BM-derived cells from donors of various ages was 5 x <em>1</em>06 cells/<em>1</em>0-cm dish. BM-derived cells were also grown in the absence of Dex at this plating density. At the optimal plating density, in the presence of Dex, the number of CFU-F/ALP+ present in the BM of the younger donors (3-36 years old) was 66.2 +/- 9.6 per <em>1</em>06 cells (mean +/- SEM), but only <em>1</em>4.7 +/- 2.6 per <em>1</em>06 cells in the older donors (4<em>1</em>-70 years old). With longer-term culture (4-5 weeks) of these BM cells in medium containing <em>1</em>0 mM beta-glycerophosphate and <em>1</em>00 microg/<em>ml</em> ascorbic acid, the extracellular matrix mineralized, a result consistent with mature osteoblastic function. These results demonstrate that the number of MSCs with osteogenic potential (CFU-F/ALP+) decreases early during aging in humans and may be responsible for the age-related reduction in osteoblast number. Our results are particularly important in that the vertebrae are a site of high turnover osteoporosis and, possibly, the earliest site of bone loss in age-related osteoporosis.