Characterization of hilar lymph node by 18F-fluoro-2-deoxyglucose positron emission tomography in healthy subjects.

One hundred and seventy-nine images were collected from healthy subjects who underwent whole-body 18F-fluoro-2-d-deosyglucose positron emission tomography (FDG-PET) for health examination. Images showing visually increased FDG uptake based on a five-point visual scale in the hilar regions were included for analyses. We evaluated the sizes, standard uptake values (SUV), and lesion-to-background (L/B) ratios of the hilar lymph nodes (HLN). Fifty of 179 (28%) subjects had visually increased FDG uptake in the hilar regions with a total of 84 HLN. By a five-point visual scale, 67 out of 84 (79.8%) HLN were grade 2, 16 out of 87 (19%) were grade 3, and 1 out of 84 (1.2%) was grade 4. The average size of the HLN was 1.55 x 1.46 cm. SUV of the HLN ranged from 1.132-2.975 with an average of 1.8 +/- 0.44. L/B ratio of the HLN ranged from 1.05-2.63 with an average of 1.52 +/- 0.39. Twenty-eight % of healthy subjects who underwent whole-body FDG-PET demonstrated visually increased FDG uptake in the HLN. However, all of the 84 HLN had a size < 2 x 2 cm, SUV < 3.0 and L/B ratios < 3.

Lumbar spine range of motion as a measure of physical and functional impairment: an investigation of validity.

OBJECTIVE: To investigate the validity of the spinal range of motion models outlined in the second and fourth editions of the American Medical Association Guides to the evaluation of permanent impairment (AMA Guides), for assessing the percentage impairment in chronic low back pain patients. DESIGN: Cross-sectional validation study. SETTING: Outpatient department in the Rehabilitation Medicine Unit. SUBJECTS: A volunteer sample of 34 subjects participated in the study, 21 females and 13 males, with a mean age of 47.7 years (1 SD = 12.1) and 40.1 years (1 SD = 11.1), respectively. Subjects had chronic low back with or without leg pain of at least six months' duration. Subjects were recruited by medical practitioners and physiotherapists through the Rehabilitation Unit at the Essendon Campus of Royal Melbourne Hospital. MAIN OUTCOME MEASURES: Lower back range of motion measured with a long arm goniometer and a dual inclinometer, Waddell Physical Impairment Scale, Waddell Disability Index, Oswestry Disability Index. RESULTS: Both range of motion measurement methods demonstrated poor validity and do not bear any consistent relationship to the level of physical or functional impairment in subjects with chronic low back pain. CONCLUSIONS: There was no evidence for a relationship between low back range of motion and impairment, and thus it would appear illogical to evaluate impairment in chronic low back pain patients using a spinal range of motion model when aiming to measure or compensate disability.

Reliability of the American Medical Association guides' model for measuring spinal range of motion. Its implication for whole-person impairment rating.

STUDY DESIGN: Repeated measures design for intra- and interrater reliability. OBJECTIVES: To determine the intra- and interrater reliability of the lumbar spine range of motion measured with a dual inclinometer, and the thoracolumbar spine range of motion measured with a long-arm goniometer, as recommended in the American Medical Association Guides. SUMMARY OF BACKGROUND DATA: The American Medical Association Guides (2nd and 4th editions) recommend using measurements of thoracolumbar and lumbar range of movement, respectively, to estimate the percentage of permanent impairment in patients with chronic low back pain. However, the reliability of this method of estimating impairment has not been determined. METHODS: In all, 34 subjects participated in the study, 21 women with a mean age of 40.1 years (SD, +/- 11.1) and 13 men with a mean age of 47.7 years (SD, +/- 12.1). Measures of thoracolumbar flexion, extension, lateral flexion, and rotation were obtained with a long-arm goniometer. Lumbar flexion, extension, and lateral flexion were measured with a dual inclinometer. Measurements were taken by two examiners on one occasion and by one examiner on two occasions approximately 1 week apart. RESULTS: The results showed poor intra- and interrater reliability for all measurements taken with both instruments. Measurement error expressed in degrees showed that measurements taken by different raters exhibited systematic as well as random differences. As a result, subjects measured by two different examiners on the same day, with either instrument, could give impairment ratings ranging between 0% and 18% of the whole person (excluding rotation), in which percentage impairment is calculated using the average range of motion and the average systematic and random error in degrees for the group for each movement (flexion, extension, and lateral flexion). CONCLUSIONS: The poor reliability of the American Medical Association Guides' spinal range of motion model can result in marked variation in the percentage of whole-body impairment. These findings have implications for compensation bodies in Australia and other countries that use the American Medical Association Guides' procedure to estimate impairment in chronic low back pain patients.

Strategies for diagnosing HIV-1 infection in atypical Western blots.

The Western blot (WB) has long been used to confirm positive ELISAs for diagnosing HIV-1 infections. However, some WB patterns may result in "indeterminate" or controversial reports thus impeding early diagnoses or accurate diagnoses. The interpretation of HIV-1 WB has no "gold standard" criterion. Incomplete antibody profiles on WB strips can be interpreted as positive or indeterminate according to different criteria. The possibility of HIV-2 infection was further checked in these serum samples. However, no reactivity to synthetic peptide of HIV-2 gp36 had been found. Serial WB analyses are important for attaining early diagnoses of HIV-1 infections as well as for evaluating clinical stages. Temporal changes on WB patterns of serial serum samples provide the evidence of seroconversion in individuals with risk behaviours and indeterminate WB. In late stage of HIV-1 infection, the reactivity to gag, pol and env antigen groups may decrease and result in indeterminate WB. We propose to diagnose HIV-1 infection and to differentiate the infection of HIV-1 from HIV-2 in these cases by using nested polymerase chain reaction (PCR) to demonstrate the presence of HIV-1 specific vpu gene.

Biological characteristics of human hepatocellular carcinoma cell lines with or without prior nude mouse passages.

A cell line, J5, derived directly from the human hepatocellular carcinoma (HCC) by in vitro culture, and three other lines, J2-23, J3-27 and J3-28, which were established in culture only after passages in nude mice were examined. Since nude mice as well as various strains of normal mice often produce infectious murine retroviruses, the HCC cells passaged as a heterotransplant in nude mice may be infected with these viruses. The supernatant of the J2-23, J3-27 and J3-28 cell co-cultured with a wild mouse cell line, SC-1, which supports the murine ecotropic retroviruses, showed the presence of a murine N-tropic virus. No murine xenotropic retrovirus was detected in these cells using S+L- mink cell assay method. A clone (J3-28 Clone 1) was derived from the above mentioned J3-28 cells, since the latter contained a mixture of human and murine cells. The J3-28 Clone 1 was found to be entirely of human karyotype. This clone as well as the J5 and the PLC/PRF/5 cel lines which have never been passaged in nude mice, showed none of these murine retroviruses. By Southern-blot hybridization, no change was detectable for c-abl, c-ras, c-mos, and c-myc protooncogenes in the chromosomal DNA of J2-23, J3-27, J3-28 Clone 1, and J5 cell lines. The karyotypes of all the HCC cell lines were aneuploid, and those of the J2-23 and J3-27 cell lines were of mouse, while the J3-28 Clone 1 and J5 cell lines were of human. The chromosomes of the J2-23 and J3-27 cells showed, except for the presence of a few marker chromosomes in the former, no apparent changes in the length were observed. In contrast, those of the J3-28 Clone 1 and J5 cells exhibited various changes, including 6-8 marker chromosomes, and an increase or decrease in the length of p and/or q arms of various chromosomes.

Specificity of antinuclear antibodies in primary biliary cirrhosis.

OBJECTIVE: To study the specific autoantibodies against nuclear antigens in patients with primary biliary cirrhosis (PBC). METHODS: Sera from 21 patients with PBC were tested for antinuclear antibody (ANA) by indirect immunofluorescence on human epithelial (HEp)-2 cells, and for antibodies to various nuclear antigens by enzyme linked immunosorbent assay (ELISA) using different purified proteins or recombinant proteins as antigens. RESULTS: ANA detected in 10 of 21 patients (48%) with PBC included five anti-centromere antibody (ACA), two speckled, two homogeneous and one nuclear dot staining. ACA were present in 24% of PBC patients. By ELISA, anti-histone antibodies were detected in 81% of PBC patients, anti-ssDNA antibodies in 71% and anti-dsDNA in 10%, anti-topoisomerase-1 antibodies in 24%, anti-Sm/RNP antibodies in 24%, anti-La-48(SS-A) antibodies in 21%, and anti-Ro-60(SS-A) and anti-Ro-52(SS-A) antibodies in 30% and 25%, respectively. CONCLUSIONS: The high frequencies of various antibodies directed against intracellular proteins and nucleic acids in patients with PBC suggests that PBC is a multisystem autoimmune disease which is similar to other systemic autoimmune diseases.

Stability of HBV DNA in cell lines and nude mouse-passaged tissues derived from human hepatocellular carcinoma.

Human hepatitis B virus (HBV) infection has been closely linked to the occurrence of hepatocellular carcinoma (HCC). Hepatoma cell lines and nude mouse-passaged hepatoma tissues were used in this report to study the HBV DNA status in these cells after passage. DNA was extracted from seven hepatoma cell lines and three nude mouse passaged HCC lines. Southern blot hybridization technique was performed with either cloned HBV whole genome or subgenomic DNA fragments as probes to analyze the presence of HBV DNA. Integration of HBV DNA fragments was detected in one mouse passaged tissue, R. Hybridization with HBV subgenomic DNA revealed that there were some DNA rearrangements of the integrated HBV DNA in R. However, the integrated HBV DNA could not be detected in the cell line derived from R after in vitro cultivation for 2 years. Both episomal form and integrated HBV DNA were detected in a cell line NTU-h3. Episomal form HBV DNA ih NTU-h3 changed after several passages. HBV DNA in NTU-h3 was unstable after in vitro cultivation. Therefore, we concluded that the presence of HBV DNA might not be essential for the maintenance of the tumorigenicity of hepatoma and the nude mouse system was more stable for maintaining HBV DNA in HCC.

Expression and antigenecity of human immunodeficiency virus type-1 transmembrane protein gp41 in insect cells.

The HIV-1 transmembrane protein, gp41, is processed together with the envelope glycoprotein, gp120, from the same precursor, gp160, during the virus maturation. We used a baculovirus expression system to demonstrate that gp41 could be properly expressed without the preceding gp120 sequence. Two constructs with slight differences in the N-terminal region of gp41 were generated: one with a deletion of the first 7 hydrophobic residues of gp41, which have been suggested to be in a region important for membrane fusion and penetration, whereas the second with a complete sequence of gp41 except that a nonconserved leucine was substituted with a glutamine during DNA manipulation. Results from Western blotting with specific antisera confirm the gp41 identity. The sizes of gp41 were sensitive to tunicamycin treatment, indicating that N-linked glycosylation did occur. Further immunoblotting analyses with 90 different serum samples from HIV-1-infected individuals gave similar reaction patterns, suggesting that gp120 as well as the N-terminal region of gp41 are not critical for the expression and antigenecity of gp41. These eucaryotic constructs should provide valuable gp41 sources for detailed characterization of gp41 functions.

Infrequent mutation of p53 gene in hepatitis B virus positive primary hepatocellular carcinomas.

Somatic mutation of the p53 oncogene/anti-oncogene allele has been shown to be involved in many different human solid tumors. Recently, there have been reports that p53 mutations are found to occur at high frequency (50%) in aflatoxin-related human primary hepatocellular carcinomas (HCC) (Hsu et al., 1991 Nature, vol 350, p. 427; Bressac et al., 1991 Nature, vol 350, p. 429). Most strikingly, a hotspot G to T mutation at amino acid position 249 was identified. These reports appear to contradict our earlier publications that although p53 mutation is found frequently in human HCC cell lines, it is rarely found in primary tumors. In this paper, we have further examined 20 different primary HCC samples (17 were hepatitis B surface antigen positive) and their adjacent nontumourous tissues, using restriction fragment length polymorphism (RFLP) analyses. Clear loss of heterozygosity (LOH) was found in only 3 out of 20 samples. All three samples were also found to carry a point mutation within the remaining p53 allele. None of these mutations was found to be at the proposed aflatoxin hotspot of amino acid 249. All three point mutations are of somatic origin. Ten samples, randomly chosen from the remaining 17 LOH negative HCC tumors, were analyzed further by DNA sequencing and Western blot analyses. No point mutations of p53 were found. Taken together with our previous report (Hosono et al., 1991, Oncogene vol 6, p. 237-243), we conclude that p53 mutation occurs infrequently, only approximately 18%, in HBV-positive primary hepatomas from Taiwan. Furthermore, p53 mutation appears to be acquired later in tumor development at least in some HCC samples.

Reactivity of anti-mitochondrial antibodies in primary biliary cirrhosis and systemic sclerosis.

Anti-mitochondrial antibodies (AMA) were detected by indirect immunofluorescence in the sera of 16 out of 17 (94%) patients with primary biliary cirrhosis (PBC). Immunoblotting experiments with mitochondrial polypeptides from the porcine liver as antigens revealed that three antigens were recognized by the sera from AMA-positive patients. These were a 70-kD protein recognized by nine out of 16 AMA-positive sera, a 50-kD protein recognized by 13 out of 16 AMA-positive sera and a 39-kD protein recognized by four out of 16 AMA-positive sera. The reactivity of these polypeptides was destroyed by brief exposure to trypsin. None of these antigens were recognized by any of the 30 control sera. These results show that the 70-kD, 50-kD and 39-kD proteins are the major mitochondrial autoantigens recognized by sera from patients with PBC. In addition, of 30 sera samples from patients with diffuse scleroderma, 13 reacted to the 70-kD and/or 50-kD antigens. Anti-centromere antibodies (ACA) were also detected in the sera of five of the 17 (29%) patients with PBC. The high prevalence of ACA in patients with PBC and the presence of anti-70- and 50-kD antibodies in patients with diffuse scleroderma provide evidence of an association between these two disorders.

Molecular analysis of the p53 alleles in primary hepatocellular carcinomas and cell lines.

We have examined p53 oncogene/anti-oncogene alleles in 10 different human hepatoma cell lines and 18 primary hepatocellular carcinomas. The p53 allele in these hepatoma cell lines appears to be a frequent target of mutation as demonstrated by Southern and Northern blotting, immunoprecipitation and Western blot analysis. In general, the steady state level of p53 specific RNA or protein in these hepatoma cell lines is higher than in normal liver. However, in three out of ten cell lines, normal-sized p53 mRNA cannot be detected. In contrast, the involvement of the p53 allele in primary hepatocellular carcinoma appears to be an exceedingly rare event. Steady state levels of p53 specific RNA in primary hepatomas are practically indistinguishable from those in normal adult liver. Using the polymerase chain reaction technique, we have amplified and subcloned exons 5, 6, 7 and 8 of p53 from 10 different hepatoma samples. DNA sequence analysis of these exon subclones reveals no apparent structural alterations. Finally, synthesis of p53 specific mRNA or protein in a HepG2 human hepatoblastoma cell line does not appear to be affected by gene expression and replication of human hepatitus B virus. Surprisingly, unlike many other kinds of human solid tumors, point mutations in p53 do not appear to be important in primary tumors of hepatocellular carcinomas.

Application of a procedure starting with an HIV-I/HIV-2 mixed EIA.

A procedure to diagnose the infections by the human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2) was proposed. The specimens were first screened by a mixed enzyme immunoassay (EIA) for the presence of antibodies to HIV-1 as well as to HIV-2. Those screened positive were thereafter confirmed and differentiated their antibody reactivities toward the antigens of HIV-1 and HIV-2 by Western blot (WB). This procedure was found to be one hundred percent accurate to diagnose 66 coded specimens with well defined seroreactivities to HIV-1 from Centers for Disease Control (CDC). While its accuracy in HIV-2 antibody testing could not be evaluated in the present study owing to the lack of HIV-2 standard reference specimens. With this procedure, six out of each of two groups of 176 foreigners and 719 individuals who visited the Taipei Municipal Venereal Disease Control Institute (TMVDCI) were screened repeatedly reactive by the mixed EIA. By WB only four of the first six and two of the second six were confirmed HIV-1 positive. Their antibody reactivities toward HIV-2 antigens were either absent or significantly much lower than those toward HIV-1 antigens. Furthermore, none reacted to the env proteins gp160 and gp120 of HIV-2. We therefore concluded that out of the 895 test individuals, only six were confirmed infected with HIV-1. There were neither HIV-2 infections nor dual infections. The similar pathogenicity and trans mission of HIV-2 as of HIV-1 justify an equal appreciation for HIV-2 testing. The present study indicated that a procedure starting with a mixed EIA aimed to diagnose HIV-1 as well as HIV-2 did not have any adverse effects on HIV-1 testing.

Antigenic characterization of the human immunodeficiency viruses.

As more is learned about the human immunodeficiency viruses HIV-1 and HIV-2, increasingly sophisticated methods of acquired immunodeficiency syndrome (AIDS) treatment and prevention may be implemented. Integral to an understanding of these viruses is an analysis of both the viral antigens and the host-immune responses to these antigens, which may differ from HIV-1 to HIV-2. Because levels of both antigen and antibody vary throughout disease development, knowledge of how and why such changes occur will lend insight into viral pathogenic mechanisms and will facilitate the development of differential diagnostic tests for classifying AIDS patients and their disease states. This task becomes very complex when dealing with HIV viruses because they possess an unprecedented number of regulatory genes for members of the retrovirus family.

Langerhans cells expressing HLA-DQ, HLA-DR and T6 antigens in normal oral mucosa and lichen planus.

We compared Langerhans cells (LC) expressing HLA-DQ, HLA-DR and T6 antigens in biopsies from the same oral mucosal site in 12 patients with oral lichen planus and eight healthy volunteers. LC expressing each antigen were observed in all the specimens, but in lichen planus the cells were located in higher levels of the epithelium than in controls. Compared with controls, lichen planus contained significantly more HLA-DQ-positive LC (P = 0.04) and fewer HLA-DR-positive LC (P = 0.05), but there was no such difference in T6-positive LC. In lichen planus specimens, there were significantly more LC expressing HLA-DQ and T6 than HLA-DR (P = 0.0001 and 0.02 respectively); no such differences were found in normal mucosa. Epithelial cells in lichen planus expressed HLA-DR antigen, but not HLA-DQ or T6 antigens. We conclude that in lichen planus there is modulation of HLA-DR and HLA-DQ antigen expression by LC, or differences in the number of LC expressing those antigens.

Infection of human T-cell leukemia virus type I and development of human T-cell leukemia lymphoma in patients with hematologic neoplasms: a possible linkage to blood transfusion.

Among 354 adult patients with either hematological malignancy or aplastic anemia, eight were positive for anti-HTLV-I antibodies; six of eight had received multiple transfusions. There was an approximately 3.5-fold increase (P less than .001) of HTLV-I seropositivity in the patients with hematologic disease (8 of 354, 2.23%) compared to the healthy adults older than 20 years (34 of 5252, .65%). Two hematological patients, one with Hodgkin's disease and one with acute promyelocytic leukemia, were found to be positive for HTLV-I, and developed and died of adult T-cell leukemia/lymphoma (ATL) subsequently. Both were long-term survivors of the primary disease and had received multiple transfusions. The latent period from blood transfusion to onset of ATL was 6 months and 11 years, respectively. Immunocompromised patients, who were seropositive for HTLV-I, may be at increased risk for ATL compared to healthy carriers of HTLV-I, and the latent period may be shorter.

High prevalence of human papillomavirus infection and possible association with betel quid chewing and smoking in oral epidermoid carcinomas in Taiwan.

Seventeen oral epidermoid carcinomas, three oral papillomas, and 17 normal gingival tissues were tested for the presence of human papillomavirus (HPV) types 6, 11, 16, and 18 sequences by Southern blot hybridization. Episomal HPV-16 sequences in various amounts were detected in 76.4% of the oral carcinomas and in all three cases of papilloma. However, only one of the 17 normal tissues was HPV positive with an unknown type. None of the samples contained HPV-6, -11, or -18 sequences. Examination of the habits of the patients showed that 59% of the patients were betel quid chewers and 82% were smokers. Thus, the concurrent incidence of HPV infection and betal quid chewing and/or smoking habits in oral carcinoma patients observed in Taiwan is consistent with the view that both viral and chemical factors may be involved in the process of carcinogenesis.

Molecular characterization of the SM and RNP nuclear antigens.

Patients with systemic lupus erythematosus (SLE) often possess antibodies against two nuclear antigens, designated "Sm" and "RNP". The exact relationship between Sm and RNP is not clear; the present study was conducted to define these two different nuclear antigens. Rabbit thymus extracts were used to obtain purified Sm/RNP complex and free Sm antigens by using a combination of 25-60% ammonium sulfate precipitation, DEAE-Sephacel and hydroxylapatite chromatography. By using the separated antigens, sera characterized as anti-Sm, anti-Sm/RNP and anti-RNP could be distinguished by enzyme-linked immunosorbent assay (ELISA). Anti-Sm and/or anti-RNP antibodies were detected in 32 (52%) of 62 sera from patients with SLE. Of these 32, 6 contained anti-Sm only, 10 contained anti-RNP only and 16 had both. When HeLa nuclear extracts were used as antigens by immunoblotting, sera with anti-RNP reacted primarily with 2 polypeptides of 68 and 45 KD; sera with anti-Sm recognized mainly on 2 polypeptides of 26 and 14 KD; sera with anti-Sm/RNP recognized both groups. When purified Sm/RNP complex from rabbit thymus extracts was used as antigen by immunoblotting, sera with anti-RNP reacted with 68 KD protein and putative degradation products of the 68 KD protein. (major: 63 KD, 45 KD, 40 KD; minor: 54-47 KD); sera with anti-Sm recognized 14 KD protein; sera with anti-Sm/RNP reacted with both groups. Although Sm and RNP can exist as a Multimolecular complex, the epitopes recognized by anti-Sm and anti-RNP differ greatly. The Sm determinants reside primarily on proteins of 26 KD and 14 KD, whereas the RNP determinants reside mainly on a protein of 68 KD.

Seroepidemiology of human T-cell lymphotropic virus type I infection in Taiwan.

The epidemiological characteristics of human T-cell lymphotropic virus type I infection in Taiwan have been explored by an island-wide community-based survey, which was carried out among residents in 19 townships and metropolitan precincts randomly selected through stratified sampling. Serum specimens of 7278 healthy subjects were screened by enzyme-linked immunosorbent assay and confirmed by Western blot method. A total of 103 subjects showed positive or weak reactions by enzyme-linked immunosorbent assay, but only 35 of them were confirmed to be positive by Western blot analysis. The anti-human T-cell lymphotropic virus type I antibody positive rate was 4.81/1000. The seropositive rate increased with age in both males and females, and females had a greater seropositive rate than males for all the age groups. Aborigines and Hakka Taiwanese had higher seropositive rates than Fukien Taiwanese and Mainland Chinese. Those people with lower educational levels were found to be associated with higher anti-human T-cell lymphotropic virus type I seropositive rates.

Human immunodeficiency virus type 1 has an additional coding sequence in the central region of the genome.

Eight coding regions designated gag, pol, env, sor, R, tat, art/trs, and 3' orf have been identified in the genome of the human immunodeficiency virus type 1 (HIV-1). Several other open reading frames have the potential to encode additional viral proteins. In this study, we show that HIV-1 has another coding sequence whose product is expressed during natural infection. Unlike antibody to other HIV-1 proteins, the prevalence of antibody to the product encoded by this region is elevated in patients with acquired immune deficiency syndrome (AIDS). Because no analogous coding region has been identified in HIV-2, the antibody to the product of this coding region may serve as a marker to distinguish infection with HIV-1 from infection with HIV-2.