Shanker Japa

HTLV-I and HTLV-II virus expression increase with HIV-1 coinfection.

Coinfections with HIV-1 and HTLV-I or HTLV-II have been associated with unique immunophenotypes and an increased risk for development of neurodegenerative conditions. These findings may result from an increased HTLV-I or II viral burden in dually infected individuals. To investigate this possibility, HTLV-I/II tax/rex messenger RNA and viral antigen expression in peripheral blood mononuclear cells (PBMCs) were measured in 37 HTLV-I- or HTLV-II-infected subjects with or without HIV-1 coinfection. Tax/rex messenger RNA was detected in 14 of 24 PBMC samples from dually infected subjects, compared with only 1 of 13 PBMC samples from singly infected subjects (58% versus 7%; p < .003). The reverse transcription-polymerase chain reaction (RT-PCR) assay correlated with HTLV-I/II viral antigen detection in PBMC cultures but not with HIV-1 viral load levels in plasma. These findings may provide clues regarding the pathophysiologic consequences of HIV/HTLV-I and HIV/HTLV-II coinfections.

Tropical Spastic Paraparesis/Human T Leukemia Virus Type 1—Associated Myelopathy in HIV Type 1—Coinfected Patients

BACKGROUND Tropical spastic paraparesis/human T leukemia virus type 1 (HTLV-1)-associated myelopathy (TSP/HAM) is rarely reported in the United States. The causative agents of TSP/HAM are HTLV-1 and, possibly, its cosmopolitan variant, human T leukemia virus type 2 (HTLV-2). Among HTLV-1- or HTLV-2-monoinfected individuals, the estimated lifetime risk for development of TSP/HAM is <2%. However, it has been suggested that HIV/HTLV coinfection may increase the risk for development of TSP/HAM. METHODS A total of 2239 human immunodeficiency virus (HIV)-infected patients were tested for HTLV-1 and HTLV-2 infection at the New Orleans Outpatient Clinic (Louisiana) during the period 1991-1998. HTLV-1-infected patients with suspected myelopathy were referred for additional evaluation. RESULTS Four cases of TSP/HAM (9.7%) were identified among 41 individuals with Western blot-confirmed HTLV-1 infection. The diagnosis was confirmed with use of molecular diagnostic assays and viral isolation. No TSP/HAM cases were identified among 65 patients with HIV-HTLV-2 coinfection. An additional patient with HIV-HTLV-1 coinfection also received a diagnosis of TSP/HAM at the New Orleans Veterans Affairs HIV Outpatient Clinic (Louisiana). All patients had normal CD4+ T cell counts at the time of diagnosis. CONCLUSIONS Given the high rates of HIV-HTLV coinfection in the United States, a heightened suspicion for TSP/HAM should be considered in HIV-infected patients who present with normal CD4+ T cell counts and myelopathy in the absence of other acquired immunodeficiency syndrome-defining conditions.

Immune responses induced by intranasal imiquimod and implications for therapeutics in rhinovirus infections

Notwithstanding the progress recently made in immunology and virology, there is yet no effective, specific treatment for the common cold. Symptomatic treatment is minimally effective. An anecdotal report of rapid clearing of the common cold of recent onset after intranasal application of imiquimod in several subjects by one of the authors, made us test the hypothesis that this treatment works through the secretion of interferon by the nasal mucosa. We decided to do an animal study in primates (Indian Macaca Mulata): 5 treatment and 3 control animals were used. Imiquimod or placebo was massaged into the nares of the animals and periodic samples of post‐nasal fluid were taken and measurements for Interferon α (IFNα) and Tumor Necrosis Factor α (TNFα) were made by ELISA methods, and kinetic studies. IFNα mRNA was also isolated and analyzed by quantitative competitive RT‐PCR. The internal standard was constructed to be complementary to and compete with oligonucleotide primers and for amplification of target sequences. One intranasal application of imiquimod rapidly (1–4 h) induced high levels of mRNA for IFNα, and minimal levels in the control animals. Rapid induction of INFα, and proportional increase of TNFα sustained for 4 and 6 h respectively were noted. No adverse reactions to treatment were found in macaques during this short period of intranasal imiquimod usage (except in one macaque with a short period of lacrimation). No animal had cytotoxic effects when examined at 6 h, 12 h, 24 h or 48 h, except one animal, which had an episode of lacrimation for 6hr post treatment. Thus both safety and efficacy of short treatment with imiquimod is proven in this animal model. Proof of principle for intranasal treatment of the common cold with imiquimod is shown. We think that this work will encourage a number of double blind clinical trials to confirm the effectiveness of the intranasal treatment of the common cold with imiquimod.

Effects of Riboflavin and Folic Acid Supplementation on Plasma Homocysteine Levels in Healthy Subjects

Background:Observational studies have shown an inverse relationship between vitamin B2 status and total homocysteine levels, a risk factor for cardiovascular disease. We hypothesize that intervention with riboflavin will lower total homocysteine levels. The total homocysteine lowering by the three genotypes (CC, CT, TT) of methylenetetrahydrofolate reductase polymorphism (677C→T) was also studied. Methods:The decrease in total homocysteine levels after supplementation with riboflavin (10 mg/d) or folic acid (1 mg/d) for 3 weeks was compared in two groups of healthy subjects (17 per group, matched by age and gender) (Phase 1). Then, both groups received supplementation with folic acid and riboflavin for an additional 3 weeks (Phase 2). Results:During Phase 1, total homocysteine levels were lowered by 2% or 4% after supplementation with riboflavin or fatty acid, respectively, although neither decrease was statistically significant (P = 0.50 and 0.19). Compared to subjects of CC genotype, total homocysteine lowering in subjects of CT genotype was approaching significance (P = 0.059) for the folic acid group, but not for the riboflavin group. After Phase 2, total homocysteine levels were not lowered significantly in either the folic acid (1%) or the riboflavin (2%) group. However, in the folic acid–riboflavin combined group, total homocysteine lowering in subjects of TT type was larger when compared to subjects of CC and CT types (P = 0.007). Conclusions:Riboflavin supplementation did not lower total homocysteine levels in healthy subjects with CC type of C677T polymorphism. However, supplementation with folic acid or with both folic acid and riboflavin may be important for CT and TT subjects in optimizing their homocysteine metabolism. These findings are relevant in characterizing the factors controlling the high total homocysteine levels for subjects of CT and TT genotypes.

Salt Loading Increases Urinary Excretion of Linoleic Acid Diols and Triols in Healthy Human Subjects

Increased dietary linoleic acid has been associated with reduced blood pressure in clinical and animal studies possibly mediated by prostaglandins. Urinary linoleate and prostaglandin metabolite excretion were investigated in subjects exposed to a salt-loading/salt-depletion regimen. Twelve healthy subjects were recruited from the New Orleans population (before Hurricaine Katrina) and admitted to the Tulane-Louisiana State University-Charity Hospital General Clinical Research Center after a 5-day outpatient lead-in phase on a 160-mmol sodium diet. On inpatient day 1, the subjects were maintained on the 160-mmol sodium diet, and a 24-hour urine specimen was collected. On day 2, the subjects received 2 L of IV normal saline over 4 hours and continued on a 160-mmol Na+ diet (total: 460 mmol of sodium). Two 12-hour urine collections were obtained. On day 3, the subjects received three 40-mg oral doses of furosemide, two 12-hour urine collections were obtained, and the subjects were given a 10-mmol sodium diet. Urinary oxidized lipids were measured by high-performance liquid chromatography-tandem quadrupole mass spectroscopy. The excretion of the urinary linoleate metabolites, dihydroxyoctadecamonoenoic acids, and trihydroxyoctadecamonoenoic acids increased significantly during intravenous salt loading as compared with day 1 and the salt-depleted periods. The urinary excretion of 6-keto- prostaglandin F1α was unaffected by salt loading but was dramatically increased 7- to 10-fold by salt depletion. Prostaglandin E2 excretion was positively correlated with sodium excretion. The salt-stimulated production of linoleic acid diols and triols may inhibit tubular sodium reabsorption, thereby assisting in the excretion of the sodium load.

356 SALT LOADING INCREASES THE URINARY EXCRETION OF LINOLEIC DIOLS AND TRIOLS IN HEALTHY HUMAN SUBJECTS.

Background/Aims Hypertension has been associated with disturbed membrane function. Changes in membrane composition related to increased dietary intake of the polyunsaturated fatty acid linoleic acid (LOA) have been shown in clinical studies to increase red blood cell membrane LOA fraction, reduce systolic blood pressure, alter red blood cell and white blood cell sodium transport processes, and increase prostaglandin production and sodium excretion. We studied the effect of salt loading on the excretion of linoleic acid diols and triols to determine if oxidative LOA metabolism is associated with salt handling by the kidney in human subjects. Methods Twelve healthy subjects were recruited from the New Orleans population (pre-Katrina) and admitted to Tulane-LSU Charity Hospital GCRC after a 5-day outpatient lead in phase on a 160 mmol sodium (Na) diet. On inpatient day 1 (D1), the subjects were maintained on the 160 mmol Na diet and 24-hour urine was collected for the LOA diols and triols: 9,10-DiHOME, 12,13-DiHOME, 9,10,13-TriHOME, and 9,12,13-TriHOME. On day 2 (D2), the subjects received 2 L of normal saline IV over 4 hours and continued on 160 Na mmol diet (total 460 mmol Na). Two 12-hour urine collections were obtained. On day 3, the patient received three 40 mg oral doses of furosemide and two 12-hour urine collections were obtained, and the patient was given a 10 mmol Na diet. The urinary LOA metabolites were measured by HPLC-tandem quadrapule mass spectroscopy at UC-Davis. Repeated measures ANOVA with a Tukey test was used to compare LOA metabolite levels during salt-loaded and salt-depleted periods. Results The urinary excretion of all LOA diols and triols measured increased significantly three- to fivefold during the day of the intravenous salt loading (D2) when the compared with the salt-depleted periods and D1. Conclusions Salt loading stimulates the renal production of LOA diols and triols, which may inhibit tubular sodium reabsorption, thereby assisting the excretion of the sodium load.

PIII-21Prevalence of NEDD4L cryptic splice site polymorphism in African-Americans with hypertension

The ubiquitin ligase NEDD4L is a candidate gene for essential hypertension based on both physiological and genetic evidence. NEDD4L ubiquinates the distal renal tubule epithelial sodium channel (ENaC) and therefore plays an important role in regulating the trafficking of ENaC from the plasma membrane. Genetic linkage studies of hypertension have also been reported to show linkage to a region 18q which includes the NEDD4L gene. A high prevalence (30%) polymorphism in NEDD4L at G82723A has been recently identified which codes for a cryptic splice site which yields a nonfunctional protein. This polymorphism would presumably lead to an increase in the plasma membrane residence time of ENaC, enhanced distal tubule sodium and water reabsorption, and hypertension.

199 PREVALENCE OF CYP2C9, 2J2, AND SOLUBLE EPOXIDE HYDROLASE POLYMORHISMS IN AFRICAN-AMERICANS WITH END-STAGE RENAL DISEASE

CYP2C9, CYP2J2, and soluble epoxide hydrolase (sEH) have been shown to be involved in the formation and metabolism of vasoactive epoxides of the epoxygenase pathway which have been proposed to play a role in the pathogenesis of hypertension and progression of renal failure. We studied the frequency of CYP2C9*8 and *11, sEH R287Q and anR403insertion, and CYP2J2*2-*7 and the newly identified CYP2J2 polymorphisms R49S, L50L, V113M, N124S in 97 AA end-stage renal disease (ESRD) patients and 84 healthy AA to determine whether there is a significant difference in prevalence rates of these polymorphisms. The mean age of the ESRD patients 53.3 ± 12.1 years (mean ± SD) and the healthy AA was 36.8 ± 8.6 years. The ESRD patients and healthy subjects were 43.3% and 40.3% female, respectively. The DNA was isolated from peripheral blood mononuclear cells and then genotyped for the variant alleles using TaqMan-based allelic discrimination assays at the University of Washington in Seattle. The prevalence of the new CYP2J2 variant alleles ranged from 0 to 1.1% in the healthy and ESRD populations except for CYP2J2*7 allele (Table). Additional results for higher frequency alleles are shown in the table below. There was no significant difference in prevalence rates of any of the variant alleles between ESRD and healthy subjects. Table Prevalence of Epoxygenase SNPs in AA with ESRD

The prevalence of CYP2C9, 2J2, and soluble epoxide hydrolase (SEH) polymorphisms in African‐Americans (AA) on hemodialysis (HD)

CYP2C9, CYP2J2, and sEH have been shown to be involved in the formation and metabolism of vasoactive epoxides of the epoxygenase pathway which have been proposed to play a role in the pathogenesis of hypertension and progression of renal failure. The AA population with endstage renal disease (ESRD) on HD has a high prevalence of hypertension and may have altered prevalence of these polymorphisms

Prevalence of CYP2C8, 2C9, 2J2, and soluble epoxide hydolase (SEH) polymorphims in African‐Americans with hypertension (HTN)

CYP2C8, CYP2C9, CYP2J2 and sEH have been shown to be involved in the formation and metabolism of vasoactive epoxides of the epoxygenase pathway which have been proposed to play a role in the pathogenesis of hypertension.