Genotype,serotype, and phylogenetic characterization of the complete genome sequence of hepatitis B virus isolates from Malawian chronic carriers of the virus

Hepatitis B virus (HBV) genotypes have distinct geographical distribution. HBV sequences among hepatitis B carriers in Malawi have not been evaluated thus far. HBsAg serotype and genotype of HBV was determined in 20 serum samples from Malawian chronic HBV carriers, and two complete genomes and 13 entire pre-S2/S genes were sequenced directly. Genotype A HBV isolates were found in all of the samples, and serotype with adw2 and ayw2 were detected in three and 17 samples, respectively. In phylogenetic analyses, two complete genomes were classified into a subgroup A' that was described previously in South African isolates of the virus, and were separated from HBV isolates in Western countries with nucleotide differences ranging from 4.1-6.2%. The separation of subgroup A' was also evident in the tree topology of the entire pre-S1/S2, X and precore/core region, but not evident in the small-S region. The nucleotide divergences in subgroup A' were higher than those among genotype A without subgroup A' in the complete genomes as well as each of four open reading frames. All of the 13 pre-S2/S sequences were classified into the subgroup A', and clustered with known HBV isolates with ayw2 in carriers from South Africa and Zimbabwe. Three amino acids in the pre-S2/S gene were characteristic of subgroup A' with ayw2. In conclusion, unique HBV isolates of subgroup A' with ayw2 are prevalent in Malawi, and subgroup A' with a relatively higher nucleotide diversity may be a HBV isolate characteristic of the indigenous population of some African countries.     

Novel mutations in TNFRSF1A in patients with typical tumor necrosis factor receptor-associated periodic syndrome and with systemic lupus erythematosus in Japanese

Molecular defects of TNFRSF1A was investigated in members of a family presenting with typical phenotypes of tumor necrosis factor receptor-associated periodic syndrome (TRAPS) and in patients with the autoimmune disorders, systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). Genomic DNA from the members of a family with typical TRAPS, as well as from 100 patients with SLE, 100 patients with RA and 100 healthy individuals, was studied for mutations in exons 2, 3 and 4 of the TNFRSF1A gene. All individuals were Japanese. Three novel missense mutations were identified in the TNFRSF1A. The C70G mutation was identified in family members with typical TRAPS, which was the second case in eastern Asian population. In addition, the T61I and R104Q mutations were each identified in 2 of the 100 SLE patients. The T61I mutation was identified in one of the 100 healthy individuals. No mutations were identified in the 100 RA patients. Functional analysis revealed that PMA-induced shedding of TNFRSF1A from PBMCs was impaired in a patient carrying T61I. A larger scale of study will clarify whether these two mutations, T61I and R104Q, are associated with chronic inflammatory disorders, such as SLE, or not.     

Coenzyme models, 6. Addition reaction of sulfite ion to monomeric and polymeric nicotinamides. Correlation between equilibria and reaction rates

Rate and association constants (k and K) for the addition of sulfite ions to five nicotinamides substituted at the ring nitrogen, ( 2a–e ), and to poly(1-(4-vinylbenzyl)nicotinamide chloride) (poly{1-[4-(3-aminocarbonylpyridiniomethyl)phenyl]ethylene chloride}) (poly( 1 )) were determined at 30°C in aqueous systems. It was found that the reaction parameters for the addition of SO to poly( 1 ) are markedly enhanced (20500-fold in the k term and 510-fold in the K term) compared with the addition to the corresponding monomeric compound, 3-aminocarbonyl-1-benzylpyridinium chloride ( 2d ), and the enhancements are suppressed with increased ionic strengths. These enhanced reaction parameters for poly( 1 ) are deviated to the upper area by two logarithmic units from a linear log K vs. logk relationship which holds for monomeric nicotinamides. This means that the rate constant is enhanced more effectively that the association constant in the polymeric system. Plots of log K_CN^? vs. log K and of log k_{f, CN}- vs. log k- gave good linear relationships. The plot for poly( 1 ), greatly deviated again to the upper area. The SO ion interacts with poly( 1 ), a cationic polyelectrolyte, more strongly than the CN^? ion.     

HLA and hypocretin studies in Korean patients with narcolepsy

STUDY OBJECTIVES: Very few studies have evaluated narcolepsy in Asian countries, outside of Japan. Our goal was to study narcolepsy at the genetic, clinical and pathophysiological level in Korea. DESIGN: Prospective study of consecutive patients and age matched controls. Clinical data ascertained from the Stanford Sleep Inventory, Polysomnography and MSLT data, as well as clinical notes. High resolution DRB1 and DQB1 typing in all subjects and studies of CSF hypocretin-1 was also evaluated in a subset of patients. PARTICIPANTS AND SETTING: 20 patients diagnosed at St. Vincent and Korea University Hospitals (Seoul, Korea). 21 Korean control subjects. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: For narcoleptic subjects, mean age was 28.2 years old and 45% were female. Mean BMI was 23.9+/-3.4 kg/m2, a significantly higher value than that expected in an age- and sex-matched sample (p<0.01). All patients had sleepiness and cataplexy while the prevalence of other symptoms ranged from 60-75%. All but 2 subjects were HLA-DR15 (DR2), DQB1*0602 positive (90%). This high DQB1*0602 percentage compared with 24% DQB1*0602 positivity in 21 control Koreans. Protective effects were observed for the DQB1*0601 and DRB1*0406 alleles, Hypocretin (orexin) CSF studies were also performed in 6 cataplectic subjects, all of which had undetectable CSF hypocretin levels. Two of these subjects had started narcolepsy less than 1 year before analysis yet had undetectable hypocretin levels. CONCLUSION: These results illustrate the similarity of narcolepsy-cataplexy in Korea in comparisons with other more studied populations. We also identified a new potential HLA protective subtype, HLA-DRB1*0406.     

Genetic diversity of HIV type 1 in rural eastern Cameroon

To monitor the presence of genotypic HIV-1 variants circulating in eastern Cameroon, blood samples from 57 HIV-1-infected individuals attending 3 local health centers in the bordering rural villages with Central African Republic (CAR) were collected and analyzed phylogenetically. Out of the 40 HIV-1 strains with positive polymerase chain reaction (PCR) profile for both gag and env-C2V3,12 (30.0%) had discordant subtype or CRF designation: 2 subtype B/A (gag/env), 1 B/CRF01, 2 B/CRF02, 1 CRF01/CRF01.A, 2 CRF11/CRF01, 1 CRF13/A, 1 CRF13/CRF01, 1 CRF13/CRF11, and 1 G/U (unclassified). Twenty-eight strains (70.0%) had concordant subtypes or CRF designation between gag and env: 27 subtype A and 1 F2. Out of the remaining 17HIV-1 strains negative for PCR with the env-C2V3 primers used, 10 (58.8%) had discordant subtype or CRF, and 7 (41.2%) had concordant one based on gag/pol/env-gp41 analysis. Altogether, a high proportion (22/57, 38.6%) of the isolates were found to be recombinant strains. In addition, an emergence of new forms of HIV-1 strains, such as subtype B/A (gag/env), B/CRF01 and B/CRF02, was identified. The epidemiologic pattern of HIV-1 in eastern Cameroon, relatively low and high prevalence of CRF02 and CRF11, respectively, was more closely related to those of CAR and Chad than that of other regions of Cameroon, where CRF02 is the most predominant HIV-1 strain. These findings strongly suggest that this part of Cameroon is a potential hotspot of HIV-1 recombination, with a likelihood of an active generation of new forms of HIV-1 variants, though epidemiologic significance of new HIV-1 forms is unknown.     

Pulmonary complications in patients with adult T-cell leukemia

A survey of 360 patients with various hematologic diseases revealed a high frequency of respiratory complications in patients with adult T-cell leukemia (ATL) compared to others. Among 29 patients with ATL, pulmonary complications were seen in 26 patients; leukemic pulmonary infiltration in 13, bleeding in 1, interstitial pneumonitis in 1, and pulmonary infection in 13. The incidence of Pneumocystis carinii and bacterial pneumonias were high despite adequate neutrophil count. Even in chronic and smoldering ATL, respiratory diseases were found in high frequency. Many of those were leukemic cell infiltration. In ten of 13 patients with pulmonary infiltration it occurred in the early stage and 6 of them were diagnosed as having "chronic lung disease" before the diagnosis of ATL. Its histology was accompanied by fibrosis in greater or lesser degree in almost all cases. Transbroncheal lung biopsy (TBLB) is of value in diagnosis (8 of 13 cases).     

Age-related changes in the response to vasoconstrictor and dilator agents in isolated beagle coronary arteries

Summary Responses to vasoactive agents were compared in helical strips of coronary arteries isolated from beagles of 30 days, 3 months, 2 years and 12 years old. Serotonin contracted the arterial strips dose-dependently, the contraction being greater in the arteries of proximal portion than in the distal arteries. The contraction increases with age from 3 months to 12 years, although EC50 values did not differ. Angiotensin II contracted distal coronary arteries to a greater extent than the proximal ones. Age did not alter the peptide-induced contraction. In prostaglandin F₂-contracted coronary arteries, acetylcholine-induced relaxations, dependent on endothelium, were less in the arteries from senescent beagles than in those from adult beagles (2 years old). Histamine relaxed the infant beagle arteries to a lesser extent than the adult and senescent beagle arteries. Histamine-induced relaxations were attenuated selectively by cimetidine. Relaxations caused by adenosine and prostaglandin I₂ did not differ in coronary arteries from beagles of different ages. It may be concluded that greater responsiveness to serotonin of senescent beagle coronary arteries is due preferentially to increased function of serotonergic receptors rather than impaired function of the arterial endothelium responsible for the release of relaxing factor(s), although some impairment of the function is supposed, on the basis of interferences with acetylcholine-induced relaxation in the aged beagle arteries. Histaminergic H₂ receptor function appears to develop in beagle coronary arteries until 3 months of age. Send offprint requests to N. Toda     

Kinetics of hepatic transport of 4-methylumbelliferone in rats. Analysis by multiple indicator dilution method

Hepatic elimination of 4-methylumbelliferone (4MU), which has been used as a model compound for conjugative metabolism, was studied by means of a multiple indicator dilution (MID) method in the isolated perfused rat liver. Using this method, three intrinsic hepatic clearances, CL _int,inf , CL _{int,eff, and} CL _{int,seq, which represent the influx, efflux, and sequestration processes, respectively, were obtained. When the dose was increased from a low dose (50 g/rat liver) to a high dose (3000 g/rat liver), the hepatic availability of 4MU increased from 0.11 to 0.73. With increasing dose, the} CL _{int,eff value increased approximately two times, while the} CL _{int,seq value decreased to approximately one-third. The remarkable dose dependence of hepatic availability was due to nonlinearity in both} CL _{int,eff and} CL _{int,seq values. However, the} CL_int,inf value was almost independent of dose. The dose-dependent change in CL_int,seq might be explained by the saturation of conjugative metabolism of 4-MU, while the increase in the CL _{int,eff value with increasing dose might be partly explained by the nonlinear tissue binding of 4-MU, since the tissue unbound fraction determined by an ultrafiltration method using liver homogenate increased approximately 1.5 times at higher concentration of 4-MU compared to that at lower concentrations. In addition, based on a comparison of the individual intrinsic clearances, i.e.}, CL _int,inf , CL _{int,eff, and} CL _{int,seq, the major determining process of the apparent hepatic intrinsic clearance of 4MU is thought to be the sequestration process at the high dose. However, at the low dose, the membrane transport process (influx and efflux processes) as well as the sequestration process also determine the apparent hepatic intrinsic clearance}.