Human immunodeficiency virus infection as risk factor for mother-to-child hepatitis C virus transmission; persistence of anti-hepatitis C virus in children is associated with the mother''s anti-hepatitis C virus immunoblotting pattern

Furosine, formed by hydrolysis of 1-deoxy-fructosyl-lysine (fructose-lysine), is a product of the Amadori rearrangement of glucose and epsilon-NH2-lysine. Fructose-lysine can react further with tissue and circulating proteins to produce advanced glycation end-products (AGEs). Peritoneal dialysate used in the treatment of patients with end-stage renal failure contains high concentrations of glucose which may lead to intraperitoneal formation of AGEs. To quantitate the kinetics of formation and peritoneal clearance of glycated peritoneal dialysate proteins, we developed an effective approach to the measurement of furosine in clinical samples of serum and peritoneal dialysate.     

Further study of hepatitis C virus RNA detection in formalin-fixed, paraffin-embedded liver tissues by ligation-dependent polymerase chain reaction

AIM: Refractive cataract surgery using corneal incisions is aiming at neutralization of preoperative astigmatism. PATIENTS AND METHODS: 61 patients with preoperative astigmatism of 2.25 +/- 0.98 were included in the treatment. A self-sealing corneal tunnel incision measuring 4.0 to 4.1 mm in external diameter and 6.5 to 7.0 mm in internal diameter (stretch incision) was performed on the steeper axis. After capsulorhexis and phacoemulsification a 5 mm PMMA lens was implanted without suturing. Keratometry and corneal topography were performed preoperatively, 3 days and 1 year respectively following surgery. The statistical analysis was based on the Wilcoxon signed ranks test. RESULTS: Surgical induced astigmatism (IA) following superior incisions in cases of astigmatism with the rule (n = 29) amounted to 1.93 +/- 0.97, while lateral incisions in cases of astigmatism against the rule (n = 29) led to an IA of 1.35 +/- 0.73. Axial shifts by more than 30 degrees were 23% following superior incisions and 17%, after lateral incisions. We observed. astigmatic reduction of 1.3 D after superior incisions and 0.7 D following lateral incisions. CONCLUSION: By 4 mm corneal cataract incisions on the steeper axis a high preoperative astigmatism can be reduced significantly without additional keratotomies.     

Detection of hepatitis C virus with RNA polymerase chain reaction in fulminant hepatic failure

The role of hepatitis C virus (HCV) infection in fulminant hepatic failure is controversial. The frequency of serum HCV RNA positivity in previously reported patients with fulminant hepatic failure (FHF) of indeterminate cause ranged from 0 to 12% in the United States and Europe and from 43% to 59% in Asia. We assessed serum HCV RNA using polymerase chain reaction (PCR) and oligoprimers from the 5'UTR of the HCV genome in 26 consecutive patients with FHF. Another laboratory independently performed PCR on 21 of the serum samples using different oligoprimers from the 5'UTR and NS3 region of the HCV genome. Serum HCV RNA was detected in two of seven (28%) patients with hepatitis B, 9 of 15 (60%) with an indeterminate cause, and in none with hepatitis A (n = 2) or drug-induced hepatotoxicity (n = 2). HCV RNA PCR results were concordant between both laboratories in 17 of 21 (81%) of samples. In patients with an indeterminate cause, HCV RNA positivity was significantly associated with the transmission risk factor of low socioeconomic status and Hispanic ethnicity. Eighteen patients underwent liver transplantation (LT) and 15 (83%) survived. Among patients with FHF of indeterminate cause, recurrent or acquired HCV infection after transplantation occurred in three of five (60%) and one of four (25%) patients, respectively. Three of four (75%) patients with hepatitis C virus infection post-LT also developed histologic hepatitis. HCV appears to be the causative agent of a substantial number of cases of FHF classified as indeterminate in the Los Angeles area. Differences in patient populations or risk factors may explain the discordant incidences of HCV infection in FHF observed among different programs.     

A comparative evaluation of the sensitivity of five anti-hepatitis C virus immunoblot assays

The profiles, after digestion with ApaI or NotI and pulsed-field gel electrophoresis (PFGE), of genomic DNA from 18 strains of Taylorella equigenitalis isolated in Ireland were of three different types. The 13 strains in one of these types gave PFGE profiles identical to that of an American prototype strain, Kentucky 188, but different from strain NCTC11184T and from a strain isolated in Japan. Eight additional strains isolated in the United States gave four distinct types of genomic PFGE profiles. All four types were different from that of T. equigenitalis NCTC11184T or that of the Japanese strain. The profile of three strains of one type was identical to that of Kentucky 188.     

Serological diagnosis of hepatitis C virus in patients with liver disease in Saudi Arabia. Evaluation of antibody determination by recombinant immunoblot assays in relation to RNA detection by polymerase chain reaction

Sera from 164 Saudi Arabian patients with non-A, non-B hepatitis liver disease were examined for antibodies to hepatitis C virus (HCV) by second- and third-generation recombinant immunoblot assay (RIBA-2 and RIBA-3) and for HCV RNA by polymerase chain reaction (PCR). By using RIBA-2, 92 (56.1%) were reactive, 64 (39%) were nonreactive, and 8 (4.9%) were indeterminate. By using RIBA-3, 98 (59.7%) were reactive 60 (36.6%) were nonreactive, and 6 (3.7%) were indeterminate. By using PCR, 108 (65.9%) were positive. Of the eight RIBA-2 indeterminate samples, seven became RIBA-3 reactive but PCR-positive, and one became RIBA-3 nonreactive but PCR-negative. Of the six RIBA-3 indeterminate samples, five were RIBA-2 nonreactive but PCR-positive, and one was RIBA-2 reactive but PCR-negative. From our study on Saudi patients, we conclude that RIBA-3 has slightly but not significantly improved the results of anti-HCV antibody detection, and is probably of more value to resolve those indeterminate samples by RIBA-2. Although expensive, PCR remains the most reliable HCV diagnostic method until an HCV antigen detection test is available.     

Hepatitis C virus and liver diseases

Pigmented villonodular synovitis is an uncommon synovial disease which only rarely involves the hip. In a multicenter retrospective study, we identified 58 histologically-proven cases. There were 33 females and 25 males. Mean age at diagnosis was 38 years. In all but two cases, only one hip was involved; the right hip was affected somewhat more often (33 cases) than the left. Two patients probably had bilateral hip disease. Mean delay to diagnosis was four years. Pain was the presenting symptom in most cases. A palpable mass in the groin was found in six patients. Plain roentgenograms were considered normal in only three patients. Bony cysts were seen in 39 patients and kissing cysts in 19. Joint space narrowing was found in 40 patients and was diffuse in half the cases. Roentgenograms suggested pigmented villonodular synovitis in 63% of cases, osteoarthritis of the hip in 16%, and inflammatory hip disease in 14%. Additional imaging studies included opaque arthrography in 21 subjects, computed tomography in 23, magnetic resonance imaging in 11, and arthroscopy in 9. Initial treatment was osmic acid synoviorthesis in 14 patients, partial synovectomy in 9, and total synovectomy in 21; in addition, eight patients had insertion of a cup prosthesis and 13 had total arthroplasty of the hip. Treatment was successful in 65% of cases after a mean follow-up of three years; among the 35% of failures, there were seven recurrences (14%). Total hip arthroplasty was performed secondarily in nine patients. This study illustrates the diversity of roentgenological changes in pigmented villonodular synovitis of the hip and the high frequency of osteoarticular lesions precluding conservative treatment. Magnetic resonance imaging and/or arthroscopy should be used to establish the diagnosis at an early stage when conservative treatment with total synovectomy and synoviorthesis is most likely to be successful.     

Hepatitis C virus genotypes in liver transplant recipients: impact on posttransplant recurrence, infections, response to interferon-alpha therapy and outcome

BACKGROUND: End-stage liver disease due to hepatitis C virus (HCV) is the most common indication for liver transplantation in U.S. veterans. We investigated the influence of HCV genotypes on the incidence and timing of recurrent HCV hepatitis, survival, infectious morbidity, and response to interferon-alpha therapy in this unique patient population. METHODS: HCV genotype was determined by direct sequencing of the NS5 region of HCV with type-specific primers. RESULTS: Genotype 1a (66%, 32/47) was the predominant genotype. Type 1b was found in 25% (12/47) of patients and type 2b was found in 9% (4/47). Histopathologically recurrent HCV hepatitis developed in 53% (25/47) of the patients after transplantation. This group included 45% (14/31) of the patients with type 1a, 67% (8/12) of the patients with type 1b, and 25% (1/4) of the patients with type 2b (P>0.5). The time to recurrence and the severity of HCV recurrence as defined by aminotransferase levels or Knodell scores were not different among the three genotypes. There was a trend toward a higher incidence of major infections in patients with type 1b (75%) versus type 1a (48%) and type 2b (50%) (P=0.11). The response to interferon-a therapy did not differ significantly among the genotypes. Mortality at 5 years was 16% (5/31) in patients with genotype 1a, 42% (5/12) in patients with genotype 1b, and 50% (2/4) in patients with genotype 2b (P=0.06). CONCLUSIONS: The incidence, time to recurrence, and response to interferon-alpha therapy did not differ between the various genotypes in our liver transplant recipients. However, there was a trend toward higher infectious morbidity and overall mortality in patients with genotype 1b after transplantation.     

Comparison of human immunodeficiency virus 1 DNA polymerase chain reaction and qualitative and quantitative RNA polymerase chain reaction in human immunodeficiency virus 1-exposed infants

The endothelial lining of the blood-brain barrier tightly controls the distribution of peptide hormones between the central nervous system and the circulation. By using primary cultures of brain microvessel endothelial cells, an in vitro model of the blood-brain barrier, we report here the uptake and transport of the octapeptide angiotensin II by a specific receptor population. With the angiotensin II antagonists losartan (AT1 specific) and PD 123,319 (AT2 specific), we showed that both the uptake and transport of angiotensin II were mediated by the AT1 receptor. Western blot analysis confirmed the existence of the AT1 receptor in our cell-culture model. Rhodamine 123 studies also suggested that both angiotensin II antagonists, but not angiotensin II, were substrates for the P-glycoprotein efflux system, thus restricting the transport of these compounds. These results suggest an AT1 receptor mediates uptake and transport of angiotensin II at the blood-brain barrier and may contribute to the regulation of cerebrovascular levels of the peptide.     

An autopsy case of primary squamous cell carcinoma of the liver associated with hepatitis C virus antibody positive liver cirrhosis

An increase in potassium (K) intake may lower blood pressure (BP), but inconsistent results have been obtained in clinical trials. We studied the effects of K supplementation in hypertensive patients with monitoring of home and ambulatory BP. Fifty-five patients with essential hypertension (26 men, 29 women, 36-77 years old) participated in this study. A 4-week K supplementation period and 4-week control period were assigned in a randomized crossover manner. During the K period, the subjects were given 64 mmol/day of K as slow-release KCl tablets. Office, home, and 24-h BP, as well as serum and urinary electrolytes, were measured at the end of each period. In the control period, office, home, and 24-h BP were 151 +/- 2/88 +/- 1 (mean +/- SE), 138 +/- 1/83 +/- 1, and 137 +/- 1/81 +/- 1 mm Hg, respectively. Serum K increased from 4.15 +/- 0.04 to 4.42 +/- 0.05 mmol/L, and urinary K increased from 54 +/- 2 to 96 +/- 3 mmol/day with the K supplementation. Office, home, and 24-h BP were significantly lower in the K period than in the control period, although the differences were small (2.7 +/- 1.1/1.4 +/- 0.6, 3.6 +/- 0.9/1.7 +/- 0.5, 3.4 +/- 1.0/1.2 +/- 0.5 mm Hg, respectively). Changes in home and 24-h systolic BP with K supplementation were highly significant (P < .001), compared with office BP (P < .05). The change in 24-h systolic BP was correlated negatively with baseline BP and urinary Na/K ratio, and positively with baseline urinary K excretion. The changes in daytime and nighttime BP were comparable. These results indicate that increasing K intake lowers BP in hypertensive subjects, especially in those with higher BP and lower K intake. Our study supports the usefulness of K supplementation in the treatment of hypertension, although its antihypertensive effect may be small.     

Quantitative hepatitis C RNA-polymerase chain reaction and detection with DNA-ELISA

BACKGROUND/AIMS: Viral serum concentrations are considered to have a clinical, prognostic and epidemiological impact on patients with hepatitis C infection. The purpose of this study was to test whether quantitation of HCV-RNA is possible by PCR in combination with DNA-ELISA. METHODOLOGY: PCR with 25 to 35 cycles was performed with variable concentrations of cloned HCV-cDNA or the serum of patients with chronic hepatitis C. The amplified PCR-products were detected by agarose gel or by DNA-ELISA. RESULTS: The detection limit of PCR with DNA-ELISA or gel detection decreased with increasing numbers of PCR cycles. However, the correlation of the optical density of the DNA-ELISA with the HCV-cDNA concentration decreased with increasing numbers of PCR as well (r=0.8 vs. r=0.29; 25 vs. 35 PCR-cycles). HCV-RNA was found in the sera of 19 of 30 patients (63%) with chronic hepatitis C by gel detection and in 14 of 30 patients (47%) by DNA-ELISA subsequent to PCR with 35 cycles. CONCLUSIONS: The PCR/DNA-ELISA technique allows a semiquantitative determination of HCV-cDNA concentrations down to 103 genomes/ul. However, to obtain a reasonable sensitivity for HCV concentrations in the serum of patients with hepatitis C, the number of PCR cycles has to be increased to numbers too high to provide reliable quantification. Further studies should be done to evaluate whether the detection systems can be improved to obtain a sufficient sensitivity for quantitative HCV-PCR. A prerequisite for the use of PCR in combination with quantifiable detection systems is that a PCR-cycle number is chosen that keeps amplification within the logarithmic phase.     

Seroprevalence of GB virus C and persistence of RNA and antibody

Cercopithecus, the genus of guenons, is the largest of the African primate genera, and yet more than half of the species belonging to this group have never been the focus of a long-term field study. In this paper, I present data on group size and composition for a previously unstudied population of guenons on the Caribbean island of Grenada. The mona monkey, Cercopithecus mona, was introduced to Grenada from Africa approximately 200-300 years ago. Two types of social groups were found for Cercopithecus mona on Grenada: all-male groups consisting of two to four individuals and bisexual groups containing 5-32 individuals. All-male groups of Grenada mona monkeys contained any combination of juveniles, subadults, and/or adults. All-male groups were a common occurrence on Grenada but have never been reported for African C. mona and have been reported only in two other forest Cercopithecus species. Bisexual groups appeared to consist of one adult male, one to six adult females, subadult females, and juveniles and infants of both sexes. Even though no more than one adult male was ever seen in each bisexual group of monas on Grenada, other males were heard giving copulation calls simultaneously with resident adult male loud calls, suggesting that other males occasionally infiltrate bisexual groups.     

Comparison of nested polymerase chain reaction, virus isolation, and fluorescent antibody testing for identifying feline herpesvirus in cats with conjunctivitis

OBJECTIVE: To compare nested polymerase chain reaction (PCR), virus isolation (VI), and fluorescent antibody (FA) testing to detect feline herpesvirus (FHV) in cats with naturally acquired conjunctivitis or respiratory tract disease, or both. SAMPLES: Swab and microbrush specimens from the conjunctiva and throat were taken from 46 cats, allotted to 3 groups (conjunctivitis only, respiratory tract disease and conjunctivitis, and clinically normal). PROCEDURE: Cells from microbrush specimens were digested and herpesvirus DNA was amplified, using a double round of PCR. Products were detected by use of agarose gel electrophoresis. The VI and FA tests were performed in routine manner. RESULTS: Of 16 cats with conjunctivitis only, conjunctival specimens from 8 and throat specimens from 8 were FHV positive by PCR. None had positive results of VI or FA testing. Of 15 cats with respiratory tract disease and conjunctivitis, conjunctival specimens from 13 and throat specimens from 12 were FHV positive by PCR. A conjunctival specimen from 1 cat and throat specimens from 3 cats were FHV positive by VI. A conjunctival specimen from 1 cat was FHV positive by FA testing. Of 15 clinically normal cats, conjunctival and throat specimens from 2 cats were FHV positive by PCR; neither conjunctival nor throat specimens from these cats were FHV positive by VI or FA testing. CONCLUSION: For cats with respiratory tract disease and conjunctivitis, or with conjunctivitis only, nested PCR was more sensitive at detecting FHV than was VI or FA testing. CLINICAL RELEVANCE: Nested PCR is a more sensitive test than the currently available VI and FA tests for identifying FHV in cats with conjunctivitis.