bluuebunny/crossref_metadata_2025 huggingface dataset:
Renal sodium retention complicating alcoholic liver disease: Relation to portosystemic shunting and liver function
Authors: Rector William G. , Lewis Frederick , Robertson Alastair D. , Everson Gregory T.
Published: March 2007

The aim of this study was to determine whether liver function and portosystemic shunting are related to renal sodium retention in alcoholic liver disease. Twenty-three studies were performed; 10 patients had ascites. Liver function was assessed from the plasma elimination rates of antipyrine, caffeine and stable isotopes of cholic acid, the latter administered both orally [2,2,4,4-2H] and intravenously [24-13C]. Portosystemic shunt fraction was calculated as the ratio of the intravenous and oral clearances of the isotopes of cholic acid.

Portosystemic shunt fraction was similar in patients with and without ascites (61% ± 16% vs. 64% ± 11%) and unrelated to urinary sodium excretion in patients with ascites (r = −0.145). Patients with ascites had significantly lower elimination rates of all administered compounds as compared with patients without ascites (antipyrine = 0.012 ± 0.007 vs. 0.031 ± 0.016/hr, p < 0.001; caffeine = 0.014 ± 0.013 vs. 0.061 ± 0.041/hr, p < 0.002; intravenous cholic acid = 1.355 ± 0.442 vs. 2.284 ± 0.885/hr, p = 0.005; orally administered cholic acid = 2.178 ± 0.841 vs. 4.056 ± 1.837/hr, p = 0.007). However, urinary sodium excretion in patients with ascites was not related to the elimination constants of these compounds (r = 0.360, 0.319, 0.067, -0.073, respectively).

Ascites complicating alcoholic liver disease is associated with impaired liver function but not the extent of portosystemic shunting. (Hepatology 1990;12:455-459).
Acetaldehyde selectively stimulates collagen production in cultured rat liver fat-storing cells but not in hepatocytes
Authors: Moshage Han , Casini Alessandro , Lieber Charles S.
Published: March 2007

Hepatocytes and fat-storing cells have been implicated in the production of collagen, under both normal and pathological conditions. In this study, short-term primary cultures of rat hepatocytes, maintained in a serum-free, hormonally defined medium without dexamethasone and cultured on a fibronectin-collagen type IV substratum, were used. Primary and passage 1 and 2 cultures of fat-storing cells maintained on tissue culture plastic were also studied. Hepatocytes produced significant amounts of collagen type III, but formation of collagen type I was not detectable. Laminin and collagen type IV production were very low. Hepatocytes maintained their ability to metabolize ethanol (at levels comparable to those observed at 2 hr) for at least 48 hr after plating and this metabolism was inhibited 86% to 95% by 4-methylpyrazole (1 mmol/L). Neither ethanol (50 mmol/L) nor acetaldehyde (175 μmol/L, initial concentration) had any effect on the production of collagen type III or laminin. Fat-storing cells (95% to 100% desmin-positive) produced significant amounts of both type I and type III collagen. Production of collagen type IV and laminin was very low. Metabolism of ethanol by these cultures was not detected. Addition of ethanol had no effect on collagen or laminin production in fat-storing cells. In contrast, acetaldehyde significantly increased the production of collagen type I, but did not alter the production of collagen type III, IV or laminin. Incorporation of 3H-proline into total protein was not affected by addition of ethanol or acetaldehyde to fat-storing cells or hepatocytes. Exposure of fat-storing cells to ethanol or acetaldehyde did not change 3H-collagen degrading activity in the media. We conclude that fat-storing cells are likely effector cells in the increased production of collagen type I in alcoholic liver fibrosis. (Hepatology 1990;12:511-518).
Modulation of extracellular matrix proteins in rat liver during development
Authors: Reif Shimon , Terranova Victor P. , El-Bendary Mamdouh , Lebenthal Emanuel , Petell James K.
Published: March 2007

The expression and localization of extracellular matrix proteins in rat liver was investigated as a function of liver development. Levels of extracellular matrix proteins were measured by dot-blot or immunoblot protocols using monospecific antibodies prepared against collagen types I, III and IV; laminin; fibronectin; and fibronectin receptor. Proline and hydroxyproline levels from extracted liver collagen were quantitated by Pico Tag analysis. It was observed that the content of type IV collagen and fibronectin in the rat liver increased two to four times during the perinatal period. In contrast, levels of laminin and collagen types I and III decreased up to twofold in developing rat livers. The content of fibronectin receptor during ontogeny was decreased four times in an inverse relationship to fibronectin molecules. Fibronectin receptor and extracellular matrix proteins displayed no difference in apparent molecular weight as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis immunoblots. Indirect immunofluorescence staining of frozen thin liver sections revealed that the pattern of localization of extracellular matrix proteins in the nonvascular regions of fetal liver was punctate rather than restricted to a specific region such as the perisinusoidal area of adult livers. Similarly, fibronectin receptor was also present, mainly in the sinusoidal area of adult livers, whereas fetal sections were diffusely stained. Our findings suggest that the differential modulation of extracellular matrix proteins and their localization in the developing rat livers undergo a dramatic alteration in the composition and structural organization of matrix material, which may act to modulate proliferation and to promote the differentiation of liver cells during development. (Hepatology 1990;12:519-525).
Pretranslational and posttranslational regulation of the EGF receptor during the prereplicative phase of liver regeneration
Authors: Johansson Staffan , Andersson Niclas , Andersson Göran
Published: March 2007

We studied the regulation of the epidermal growth factor receptor mRNA and the number of epidermal growth factor binding sites in subcellular compartments involved in the biosynthesis and endocytosis of the epidermal growth factor receptor during the prereplicative phase of liver regeneration. The epidermal growth factor receptor mRNA, quantified by solution hybridization, decreased after partial hepatectomy, with a nadir of about 35% 18 hr after hepatectomy. An even strongèr decrease in the number of epidermal growth factor binding sites after partial hepatectomy was observed in a Golgi-enriched low-density membrane fraction, reflecting available newly synthesized epidermal growth factor receptors. It is suggested that this decrease in newly synthesized available epidermal growth factor receptors is caused primarily, but not entirely, by decreased epidermal growth factor receptor mRNA levels and the additional downregulation of epidermal growth factor binding sites may involve posttranslational mechanisms such as intracellular occupation by transforming growth factor-α.

The observation that the number of specific epidermal growth factor binding sites after partial hepatectomy was only moderately reduced in prelysosomal endosomes and in lysosomes, compared with the newly synthesized receptors, may indicate that a pool of receptors targeted for lysosomes exists and these receptors are regulated in a different manner than the receptor pool targeted for the cell surface.

Furthermore, at least two separable endocytic subcompartments are involved in the transport of the epidermal growth factor/epidermal growth factor receptor complex in the liver. The complex first enters early endosomes, then enters late, prelysosomal endosomes, where the epidermal growth factor is proteolytically processed, before arriving in the lysosomal compartment. (Hepatology 1990;12:533-541).
Changes in serum and hepatic polyamine concentrations after 30%, 70% and 90% partial hepatectomy in rats
Authors: Minuk Gearld Y. , Gauthier Tony , Benarroch Abraham
Published: March 2007

Polyamines have been reported to play an important role in stimulating hepatic regeneration after partial hepatectomy. To determine whether changes in systemic or hepatic polyamine concentrations correlate with the extent of the regenerative stimulus, serum and tissue putrescine, spermidine and spermine concentrations were determined in groups of adult male rats (n = 6 to 12/group) 0, 24, 48, and 72 hr after 30%, 70% or 90% partial hepatectomy. Serum putrescine levels were variably increased after partial hepatectomy and did not correlate with hepatic regenerative activity. Serum spermidine levels remained unaltered and spermine levels were undetectable both before and after partial hepatectomy. In hepatic tissue, only putrescine concentrations increased in proportion to the extent of hepatic resection. Moreover, there was a significant correlation between hepatic putrescine concentrations and restitution of liver mass (r = 0.778), DNA synthesis (r = 0.8026) and protein synthesis (r = 0.7034) (p < 0.05). No significant correlations existed between hepatic spermidine or spermine concentrations and these parameters of hepatic regeneration. The results of this study support the hypothesis that hepatic putrescine plays an im portant role in stimulating hepatic regeneration after partial hepatectomy. (Hepatology 1990;12:542-546).
Evidence for modulation of hepatic mass by estrogens and hepatic “feminization”
Authors: Van Thiel David H. , Stauber Rudolf E. , Gavaler Judith S. , Rosenblum Elaine
Published: March 2007

Animals with end-to-side portacaval shunts and sham-operated animals, wherein the body weight and liver weight of the animals varied spontaneously over a considerable range, were studied. The relationships between hepatic androgen- and estrogen-receptor content, serum testosterone and estradiol levels and hepatic mass were characterized. Animals with portacaval shunts were smaller than those without shunts. Moreover, they had reduced serum levels of testosterone and estradiol. The reduction in serum testosterone was greater than that of estradiol. As a result, the calculated estrogen/testosterone ratio of the two groups of animals was greater for the portacaval shunt animals than for the controls. The dissociation constant values for the androgen receptor and estrogen receptor in the liver did not differ between groups. The activity of the androgen receptor (p < 0.01) and estrogen receptor (p < 0.05) was reduced markedly in the animals with portacaval shunts compared with controls. Moreover, the hepatic cytosolic estrogen receptor activity-but not that of the androgen receptor-correlated with the measured hepatic mass in both groups of animals. These data suggest that hepatic feminization is either associated with or is a hepatic regenerative signal in the rat. (Hepatology 1990;12:547-552).
Interferon and chronic non-A, non-B hepatitis: Whom are we treating?
Authors: Conn Harold O. , Koretz Ronald L.
Published: March 2007

Chronic hepatitis C (non-A, non-B hepatitis) is a common and often progressive viral liver disease. To assess the efficacy of therapy with the antiviral agent interferon alfa, we randomly assigned 166 patients with chronic hepatitis C to treatment with either 3 million or 1 million units of recombinant interferon alfa three times weekly for 24 weeks, or to no treatment.

The probability of normalization or near normalization of the serum alanine aminotransferase levels after six months of interferon therapy was 46 percent in patients treated with 3 million units of interferon (P <0.001) and 28 percent in those treated with 1 million units (P <0.02), but only 8 percent in untreated patients. The serum alanine aminotransferase level became completely normal in 22 of the 26 patients (85 percent) who responded to treatment with 3 million units of interferon and 9 of the 16 patients (56 percent) who responded to treatment with 1 million units. The patients who received 3 million units of interferon had histologic improvement because of the regression of lobular and periportal inflammation. Relapse within six months after the completion of treatment occurred in 51 percent of the patients treated with 3 million units of interferon and 44 percent of those treated with 1 million units.

We conclude that a 24-week course of interferon therapy is effective in controlling disease activity in many patients with hepatitis C, although relapse after the cessation of treatment is common.

Infection with the hepatitis C virus may result in chronic liver disease for which no effective therapy is now available. We studied the effects of recombinant human interferon alfa in a prospective, randomized, double-blind, placebo-controlled trial in patients with well-documented chronic hepatitis C. Forty-one patients were enrolled in the trial, 37 of whom were later found to have antibody to hepatitis C virus. Twentyone patients received interferon alfa (2 million units) subcutaneously three times weekly for six months, and 20 received placebo.

The mean serum aminotransferase levels and the histologic features of the liver improved significantly in the patients treated with interferon but not in the patients given placebo. Ten patients treated with interferon (48 percent) had a complete response, defined as a decline in mean serum aminotransferase levels to the normal range during therapy; three others had a decrease in mean aminotransferase levels of more than 50 percent. After treatment ended, however, serum aminotransferases usually returned to pretreatment levels; 6 to 12 months after the discontinuation of interferon therapy, only two patients (10 percent) still had normal values.

We conclude that interferon alfa therapy is beneficial in reducing disease activity in chronic hepatitis C; however, the beneficial responses are often transient.
Suppression of hepatic lymphokine-activated killer cell induction by murine kupffer cells and hepatocytes
Authors: Tzung Shie-Pon , Gaines Katherine C. , Lance Peter , Ehrke Jane M. , Cohen Stefan A.
Published: March 2007

Murine lymphokine-activated-killer cell activity was readily induced by culturing spleen cells with 10 U/ml of interleukin-2 for 4 days. In contrast, very little activity was generated under the same culture conditions when nonparenchymal liver cells were used as the responding cells. It was concluded that Kupffer cells produced prostaglandin and interferon α/β, which suppressed lymphokine-activated-killer induction because (a) induction of lymphokine-activated-killer activity from nonparenchymal liver cells was observed in the presence of indomethacin and anti-interferon α/β antibody; (b) when adherent nonparenchymal liver cells, primarily Kupffer cells, were removed, lymphokine-activated-killer activity could be obtained with interleukin-2 alone; (c) coculture of Kupffer cells with nonadherent nonparenchymal liver cells in a two-chambered system inhibited lymphokineactivated killer cell induction in a dose-dependent manner; (d) exogenous prostaglandin E2 and interferon α/β added at the start of culture inhibited interleukin-2—induced cytotoxicity and proliferation, whereas the other major prostaglandin species in the liver, prostaglandin D2, had little effect. These findings are distinctive with Kupffer cells because splenic macrophages did not exert such inhibition in parallel experiments. Moreover, the supernatant collected from the 24-hr culture of nonparenchymal liver cells contained greater than 20-fold more prostaglandin E2 and interferon α/β than that from culture of spleen cells. In subsequent in vivo experiments, when interleukin-2 was given intraperitoneally to mice, the combination of indomethacin and anti-interferon α/β antibody significantly enhanced lymphokine-activated-killer activity recovered from the liver. Besides Kupffer cells, it was found that hepatocytes, the major cellular component of the liver, also played an inhibitory role on lymphokine-activated-killer cell generation. A cell-free liver cytosolic extract had even more potent suppressive effect, which was partially reversed by supplementation of arginine, indicating that arginase may be one of the hepatocyte-derived immunoinhibitors. (HEPATOLOGY 1990;12:644-652).
The auditory P300 event—related potential: An objective marker of the encephalopathy of chronic liver disease
Authors: Davies Mark G. , Rowan Michael J. , Macmathuna Padraig , Keeling Napoleon P. W. , Weir Donald G. , Feely John
Published: March 2007

Recently many variants of electroencephalogramevoked responses have been studied as potential diagnostic aids in the detection and evaluation of hepatic encephalopathy. This study assesses the value of the auditory P300 event—related potential — a slow component of the auditory evoked response — as a tool in this field. Twenty-one nonencephalopathic and 12 encephalopathic (grade 1/2) cirrhotic patients and 26 controls were assessed clinically and psychometrically. Electroencephalogram spectral analysis and visual evoked response recordings were also conducted. An auditory P300 wave was elicited using the standard two-tone discrimination paradigm. The latency and amplitude of this wave were measured. The latency of the P300 was found to be significantly increased in the encephalopathic patients compared with both nonencephalopathic cirrhotic and control groups (p < 0.05). Amplitude of the wave was decreased in both nonencephalopathic and encephalopathic patients, but this was not statistically significant. This study suggests that the latency of the P300 is a good marker of grades 1 and 2 clinical hepatic encephalopathy. The delays in the P300 latency may indicate that encephalopathic patients have a deterioration of their stimulus evaluation abilities. (HEPATOLOGY 1990;12:688-694).
Endoscopic ligation of esophageal varices: An endoscopist's opinion
Authors: Tedesco Francis J.
Published: March 2007

One hundred consecutive patients with bleeding esophageal varices were treated with a new endoscopic ligating device that effects strangulation of varices using small elastic “O” rings. Treatments were continued after initial hospitalization to achieve variceal eradication. Follow-up ranged from 6 to 26 (mean: 15) months. Bleeding was controlled until discharge from hospital or death in 18 of 21 patients who were actively bleeding at index endoscopy. Overall, 26 patients died during the study, 12 during the index hospitalization. Cause of death was organ failure in 21, exsanguination in 3, and cancer in 2. Forty-one of 88 initial survivors experienced 72 episodes of recurrent bleeding (1 to 4 per patient). All but five rebleeds occurred before eradication. Sixty of 88 patients (68%) who survived index hospitalization had their varices eradicated. A median of 5 (1 to 12) treatments was required. Nine patients eventually had other forms of treatment for recurrent bleeding. Only 3 non-bleeding complications resulted from 462 endoscopic treatment sessions. We conclude that endoscopic ligation controls active variceal bleeding and eradicates varices with efficacy similar to that of sclerotherapy and with minimal risk of complications.
Markers of chronic alcohol ingestion in patients with nonalcoholic steatohepatitis: An aid to diagnosis
Authors: Fletcher Linda M. , Kwoh-Gain Ian , Powell Elizabeth E. , Powell Lawrie W. , Halliday June W.
Published: March 2007

We report here the use of the biochemical marker desialylated transferrin to aid in the diagnosis of nonalcoholic steatohepatitis. Conventional biochemical tests used for the detection of chronic alcohol consumption fail to differentiate nonalcoholic steatohepatitis patients from alcoholic subjects. In addition, even in those alcoholic subjects with alcoholic liver disease in whom biopsy has been performed, it is impossible to differentiate these two disease states on the basis of morphological examination alone. In this study we have examined two new markers of excessive alcohol intake, desialylated transferrin and mitochondrial AST in subjects with nonalcoholic steatohepatitis and in patients consuming excessive amounts of alcohol. All nonalcoholic steatohepatitis patients consumed minimal or no alcohol and were diagnosed by morphological criteria based on liver biopsy specimens. Alcoholic subjects were consuming in excess of 80 gm/day ethanol, often with clinical evidence of overt alcoholism. Control subjects included both healthy controls and patient controls with liver diseases unrelated to alcohol. The ratio of desialylated transferrin/total transferrin was elevatedonlyin patients who consumed excessive amounts of alcohol, whereas the ratio of mitochondrial AST to total AST (mitochondrial AST/total AST) was not significantly different between alcoholic subjects and patients with nonalcoholic steatohepatitis. The sensitivity and specificity for the ratio desialylated transferrin/total transferrin was 81% and 98%, respectively, whereas the sensitivity for the mitochondrial AST/total AST ratio was 92%; the specificity was only 50%, indicating that there were a large number of false-positives. All the conventional markers were less sensitive and less specific than the ratio desialylated transferrin/total transferrin. We conclude that it is possible to differentiate patients with nonalcoholic steatohepatitis from patients consuming excessive amounts of alcohol using the biochemical marker desialylated transferrin but not mitochondrial AST. Mitochondrial AST is useful only in those patients with an elevated total AST, whereas desialylated transferrin detects excessive alcohol ingestion whether an elevated AST is present or not. The ratio desialylated transferrin/total transferrin remains the best single laboratory marker for the determination of chronic excessive alcohol consumption and is a valuable aid in the diagnosis of nonalcoholic steatohepatitis. (Hepatology 1991;13:455-459.)
Alterations in nuclear scaffold constituents during carbon tetrachloride-induced liver regeneration
Authors: Clawson Gary A. , Madsen Kenneth R. , Blankenship Lori J. , Hatem Christine L.
Published: March 2007

Liver regeneration was induced in rats by treatment with CCl4, which results in substantial regenerative activity with a sharp mitotic response 2 days after intoxication. Closely paralleling the mitotic index, we observed fourfold increases in nuclear scaffold nucleoside triphosphatase, an activity thought to participate in nucleocytoplasmic RNA transport and in the 46 kD putative enzyme and its selective photolabeling. Because previous work has indicated that the 46 kD protein may be proteolytically derived from lamins A/C by cleavage at a tyrosine residue at aa376, we investigated the response of lamin A/C transcripts during this regeneration. Surprisingly, Northern blot analyses after CCl4 administration showed low levels of lamin A/C transcripts (which appeared to be predominantly poly[A]-), and we found a decrease in immunoprecipitable lamins A/C from in vitro translation of poly(A)-selected RNA. To circumvent potential problems with such analyses, we used reverse transcription/polymerase chain reaction amplification of lamin A/C transcripts from total cytoplasmic RNA. These assays showed a transient, comparatively minor increase in lamin A/C transcripts 1 day after treatment, but levels rapidly declined from 1 to 3 days and were decreased at 3 to 5 days. However, nuclear scaffold protease activity, which shows a considerable selectivity for lamins A/C and may be involved in derivation of the 46 kD protein, increased in parallel to the mitotic response and increases in nucleoside triphosphatase, as assessed using a nonspecific (Azocoll) protease assay. Assays with a specific tyrosine-containing substrate (Z-Y-Sbenzyl) showed an increase that mirrored that observed with the nonspecific substrate. Our results suggest that the increases in nuclear scaffold nucleoside triphosphatase activity attendant to nuclear replication appear to result, in large part, from activation of nuclear scaffold proteases. Furthermore, one third to one half of hepatocytes apparently regenerate without any major increase in synthesis of lamins A/C, the major structural proteins of the nuclear scaffold. (Hepatology 1991;13:515-522.)
Characterization of holmium loaded alginate microspheres for multimodality imaging and therapeutic applications
Authors: Zielhuis S.W. , Seppenwoolde J.H. , Bakker C.J.G. , Jahnz U. , Zonnenberg B.A. , van het Schip A.D. , Hennink W.E. , Nijsen J.F.W.
Published: March 2007

In this paper the preparation and characterization of holmium‐loaded alginate microspheres is described. The rapid development of medical imaging techniques offers new opportunities for the visualisation of (drug‐loaded) microparticles. Therefore, suitable imaging agents have to be incorporated into these particles. For this reason, the element holmium was used in this study in order to utilize its unique imaging characteristics. The paramagnetic behaviour of this element allows visualisation with MRI and holmium can also be neutron‐activated resulting in the emission of gamma‐radiation, allowing visualisation with gamma cameras, and beta‐radiation, suitable for therapeutic applications. Almost monodisperse alginate microspheres were obtained by JetCutter technology where alginate droplets of a uniform size were hardened in an aqueous holmium chloride solution. Ho3+ binds via electrostatic interactions to the carboxylate groups of the alginate polymer and as a result alginate microspheres loaded with holmium were obtained. The microspheres had a mean size of 159 μm and a holmium loading of 1.3 ± 0.1% (w/w) (corresponding with a holmium content based on dry alginate of 18.3 ± 0.3% (w/w)). The binding capacity of the alginate polymer for Ho3+ (expressed in molar amounts) is equal to that for Ca2+, which is commonly used for the hardening of alginate. This indicates that Ho3+ has the same binding affinity as Ca2+. In line herewith, dynamic mechanical analyses demonstrated that alginate gels hardened with Ca2+ or Ho3+ had similar viscoelastic properties. The MRI relaxation properties of the microspheres were determined by a MRI phantom experiment, demonstrating a strong R2* effect of the particles. Alginate microspheres could also be labelled with radioactive holmium by adding holmium‐166 to alginate microspheres, previously hardened with calcium (labelling efficiency 96%). The labelled microspheres had a high radiochemical stability (94% after 48 h incubation in human serum), allowing therapeutic applications for treatment of cancer. The potential in vivo application of the microspheres for a MR‐guided renal embolization procedure was illustrated by selective administration of microspheres to the left kidney of a pig. Anatomic MR‐imaging showed the presence of holmium‐loaded microspheres in the kidney. In conclusion, this study demonstrates that the incorporation of holmium into alginate microspheres allows their visualisation with a gamma camera and MRI. Holmium‐loaded alginate microspheres can be used therapeutically for embolization and, when radioactive, for local radiotherapy of tumours. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res, 2007
Polymers with tunable toxicity: A reference scale for cytotoxicity testing of biomaterial surfaces
Authors: Knetsch Menno L.W. , Olthof Nadine , Koole Leo H.
Published: March 2007

A series of copolymers, with varying ratio di‐methylamino‐ethylmethacrylate (DMAEMA) and methyl‐methacrylate (MMA), was designed as a potential scale for cytotoxicity. These copolymers were characterized for toxicity of their surface. The surfaces of washed copolymers display increasing toxicity with increasing DMAEMA content. The toxicity was observed for three different cell‐types, namely mouse fibroblasts, human endothelial cells and human osteoblast‐like cells. With an increasing toxic surface, cell growth was inhibited as was indicated by the proliferation marker Ki‐67. Staining for F‐actin revealed that with increasing DMAEMA, cells adopted a more and more round morphology, resulting in decreased surface‐contact area. Immuno‐staining for phospho‐tyrosine or vinculin demonstrated gradual loss of focal adhesions on increasingly toxic surfaces. Surprisingly loss of focal adhesions coincided with an increase in paxillin and vinculin protein, indicating cells try compensating for loss of adhesion. This series of copolymers may have potential as a cytotoxicity scale. They provoke cellular responses ranging from highly toxic to completely non‐toxic, with some showing intermediate toxicity. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res, 2007
Clinical and serologic effects of live attenuated serum inhibitor‐resistant influenza B vaccine in seronegative adults
Authors: Miller Louis W. , Togo Yasushi , Hornick Richard B.
Published: March 2007

The clinical effects, nasal and serum antibody responses, and virus excretion of a live attenuated serum inhibitor‐resistant influenza B virus vaccine, R75, was evaluated in 43 seronegative healthy adults by a random double‐blind study. Symptom responses were minimal and were not significantly different between vaccine and placebo groups. No fevers, abnormalities in physical examination or laboratory testing developed during 4 weeks of observation.

Among vaccinees, 10 (48%) developed serum hemagglutination‐inhibition (HI) antibodies, 16 (76%) developed serum neutralization (N) antibodies and 4(19%) developed nasal N antibodies. The GMT responses from study day 0 to day 28 were 4.0 to 10.4 for serum HI, 1.8 to 9.8 for serum N, and 1.0 to 1.4 for nasal N. There were no significant titer changes in the placebo group. No virus excretion was detected.

Although there are some questions concerning the relationship of antibody levels to protection, the low antibody responses in this study are an indication that R75 is not sufficiently immunogenic.
Experimental infection of chimpanzees with the Norwalk agent of epidemic viral gastroenteritis
Authors: Wyatt Richard G. , Greenberg Harry B. , Dalgard Dan W. , Allen William P. , Sly D. Lewis , Thornhill Thomas S. , Chanock Robert M. , Kapikian Albert Z.
Published: March 2007

A fecal filtrate of human origin containing the Norwalk agent of epidemic viral gastroenteritis was administered by stomach tube to chimpanzees in an attempt to induce diarrheal disease. Significant postchallenge serum antibody rises against Norwalk viral antigens were demonstrated in all animals using the techniques of immune electron microscopy and radioimmunoassay. In addition, viral antigens were detected in feces from five of nine animals using radioimmunoassay. Clinical illness characterized by diarrhea and/or vomiting did not occur. Infection was transmitted subsequently by feeding four additional chimpanzees a fecal filtrate prepared from one of the previously infected animals. Development of an antibody response in four animals and detection of viral antigen in two animals that received this passage filtrate indicated that viral replication had occurred in the absence of clinical illness. The availability of the chimpanzee as an experimental animal host susceptible to infection with the Norwalk agent should facilitate the study of epidemic viral gastroenteritis.
e‐antigen and age in acute and chronic type B hepatitis
Authors: Villarejos Victor M. , Visona Kirsten Anderson , Eduarte A. Carlos E.
Published: March 2007

The influence of e‐antigen upon the course and outcome of acute type B hepatitis was studied in a series of 202 patients. Of these, 54 (27%) had e‐Ag, detected in the serum at the same time as HBsAg during the incubation period or at onset. In 39 patients (73%) the e‐Ag disappeared within eight weeks after onset, regularly followed by clearance of HBsAg approximately four weeks later; anti‐HBs was detected shortly thereafter in 34 cases. In 15 (28%) of e‐Ag‐positive and in 4 (3%) of e‐Ag‐negative patients, HBsAg persisted for one year or longer; chronic hepatitis developed in 13 of these cases, 12 of which were e‐Ag‐positive. Among e‐Ag‐positives HBsAg persisted only in those cases in which the e‐Ag also persisted; all these were persons under 15 years of age. Transaminase and bilirubin values were equally high in e‐Ag‐positive and e‐Ag‐negative patients with resolving hepatitis, but were low from the start in those who later developed chronic liver conditions, irrespective of the presence or absence of e‐Ag. It is concluded that in e‐Ag‐positive acute type B hepatitis patients the disappearance of this antigen from the serum is a good prognostic sign, whereas its persistence beyond eight weeks, especially in young children with low transaminase and bilirubin response, signals evolution towards chronicity.
Detection of Epstein‐Barr virus antigens and antibodies by peroxidase‐labeled specific immunoglobulins
Authors: Kurstak E. , de Thé G. , van den Hurk J. , Charpentier G. , Kurstak C. , Tijssen P. , Morisset R.
Published: March 2007

Detection of the Epstein‐Barr (EBV) antigens, early antigen (EA), viral capsid antigen (VCA), and nuclear antigen (EBNA) by the indirect immunoperoxidase technique was highly sensitive. Antibody titers to EBNA, EA, and VCA were determined in more than 25 sera of patients with Burkitt lymphoma (BL), nasopharyngeal carcinoma (NPC), or normal persons. A good correlation between the titers of these antigens was obtained by the immunoperoxidase and immunofluorescence methods. The indirect (anti‐IgG) immunoperoxidase technique for the detection of EBNA is, in contrast to the indirect immunofluorescence method, highly sensitive. EBNA was associated with the chromosomes in cells arrested in the metaphase with colchicine.
Solid‐phase radioimmunoassay determination of virus‐specific IgM antibody levels in a follow‐up of patients with naturally acquired measles infections
Authors: Vuorimaa Terjo O. , Arstila Pertti P. , Ziola Barry R. , Salmi Aimo A. , Hänninen Pentti T. , Halonen Pekka E.
Published: March 2007

The question of the exact disappearance time or possible persistence of measles‐specific IgM antibodies after naturally acquired measles virus infections was studied with a sensitive solid‐phase radioimmunoassay (RIA) method. A total of 30 patients were analyzed with follow‐up times varying from 4.5 to 8 months; all were measles IgM positive in the first serum specimen obtained after the onset of rash. In 29 of 30 patients, the measles IgM declined to undetectable levels by approximately 90 days. The remaining patient developed postmeasles encephalitis, however, and was found to have a prolonged measles IgM antibody response. For comparison, the measles‐specific IgG response was also studied and was found to develop only slightly later than the IgM response, with levels then remaining high and stable up to 8 months later. Although apparent measles IgM antibodies were found in 1 of 64 nonmatched adult controls, they were due to the presence of high levels of IgM‐class rheumatoid factor. The data presented indicate that measles IgM antibodies begin to decline soon after the onset of rash and reach negative levels 1 to 3 months later; in complicated infections, however, measles IgM antibody synthesis may not terminate normally.
Diagnosis of human coronavirus infection by immunofluorescence: Method and application to respiratory disease in hospitalized children
Authors: McIntosh Kenneth , McQuillin Joyce , Reed Sylvia E. , Gardner Phillip S.
Published: March 2007

Rabbit antisera were prepared against coronavirus strains 229E and OC43 and used successfully to detect viral antigen in epithelial cells shed from the nasopharynx of symptomatic volunteers who had received coronavirus inocula three to four days before. The same serologic reagents were applied to nasopharyngeal secretion cells obtained from 106 infants and children hospitalized with respiratory tract disease and apparently not infected with conventional respiratory viruses. No coronavirus infections were detected by this method. It appears that coronavirus OC43 or 229E infections were not common in children in Tyneside hospitals during the period of study. However, fluorescence is a useful method for detection of coronavirus infections in symptomatic human subjects.
Enzyme‐immunoassay in the diagnosis of hepatitis with emphasis on the detection of “e” antigen (HBeAg)
Authors: v. d. Waart M. , Snelting A. , Cichy J. , Wolters G. , Schuurs A.
Published: March 2007

A brief survey of the application of enzyme‐immunoassay (EIA) for the detection of hepatitis B surface antigen (HBsAg), hepatitis A, and their corresponding antibodies is given. The preliminary results of a similar EIA for detection of hepatitis B‐related “e” antigen (HBeAg) and its antibody (anti‐HBe) are reported. This EIA is much more sensitive than immunodiffusion: at least 128 times for HBeAg and at least 512 times for anti‐HBe. HBsAg and its antibody do not interfere with the test. Only a few sera strongly positive for rheumatoid factor gave rise to false‐positive results, as was demonstrated by a confirmatory test.
The IgM antibody responses to the core antigen of hepatitis B virus
Authors: Cohen B. J.
Published: March 2007

Little is known about the immunoglobulin class of antibodies to HBcAg. In the present study sera containing anti‐HBc were fractionated by sucrose density‐gradient centrifugation, and all serum fractions were tested against HBcAg by immunoelectro‐osmophoresis. In addition selected fractions were examined by complement fixation test, immune adherence hemagglutination and immune electron microscopy. Anti‐HBc activity in IgG serum fractions was demonstrated by all four techniques used, but HBcAg‐specific IgM was detected only by immunoelectro‐osmophoresis and by immune electron microscopy. In acute hepatitis B, HBcAg‐specific IgM was detected for up to eight weeks after the onset of jaundice. It was also found transiently in two patients who developed chronic hepatitis B without an icteric episode and in one out of thirteen patients with HBsAg‐positive chronic liver disease, but in none of eight healthy HBsAg carriers. The results suggested that HBc Agspecific IgM is formed transiently in response to primary HBV infection but is generally undetectable in established HBsAg carriers.
A simple immunofluorescent technique for the detection of human rotavirus
Authors: Moosai R. B. , Gardner P. S. , Almeida J. D. , Greenaway M. A.
Published: March 2007

If trypsin is incorporated in the tissue culture medium it is possible to carry out a sensitive immunofluorescence assay for the presence of human rotavirus. The enhanced effect of trypsin is negated by serum. It has also been established that naturally occurring enzymes in faeces enable some virus to penetrate tissue culture cells. The role of these naturally occurring enzymes in the pathogenesis of rotavirus infection is discussed.
Evaluation of aedes albopictus tissue culture for use in association with arbovirus isolation
Authors: Ajello Clayton A.
Published: March 2007

The susceptibility and sensitivity of Aedes albopictus cell cultures to five different primary and four different low‐passage arboviruses were tested. Yellow fever, West Nile, Ilesha, eastern equine encephalitis, and Flanders viruses replicated in A albopictus tissue cultures. Replication was determined by the ability of selected tissue culture fluids to infect suckling mice, and by recovery from tissue culture fluid of progressively increasing amounts of complementfixing (CF) antigen with time. Virus persistence was demonstrated with Nodamura, western equine encephalitis, and Mayaro viruses, but multiplication was not proven; neither persistence nor multiplication was demonstrated with a Kemerovo group virus. When yellow fever and Ilesha viruses were simultaneously inoculated into A albopictus culture, CF antigen for each was consistently detected. In a more detailed comparative study of field specimens, 12 unpassaged strains of yellow fever virus were tested for infectivity in A albopictus tissue culture, Vero cells, and baby mice. Higher titers of virus were detected (0.8–2.3 log ID50 per ml) in Vero cell culture than in A albopictus tissue culture or baby mouse systems. These results suggest the feasibility of using A albopictus cells in association with the primary isolation of arboviruses.
Selective detection of IgM‐antibody against core antigen of the hepatitis B virus by a modified enzyme immune assay
Authors: Gerlich Wolfram H. , Lüer Wilfried
Published: March 2007

IgM antibody against core antigen of the hepatitis B virus (anti‐HBc IgM) was selectively determined by a new enzyme immunoassay (EIA). Microtiter plates were coated with anti‐human m̈ chain immunoglobulin. On addition of serum IgM is bound by a factor of about 4,000 more than IgG. After removing the sample, HBcAg is added to the IgM‐coated surface. Binding takes place if the IgM contained anti‐HBc and was demonstrated by the aid of a conjugate made from anti‐HBc IgG and horse radish peroxidase. Quantitation may be achieved without testing a dilution series. The assay was not disturbed by a large excess of anti‐HBc IgG in the sample and rheumatoid factor did not produce false‐positive results, provided the sample was diluted in an excess of aggregated IgG. The diagnostic relevance of the assay was demonstrated in selected cases of acute hepatitis B. Rapid diagnosis of acute hepatitis B infection is therefore now possible in those cases which are HBsAg‐negative but anti‐HBc‐positive.
Dane particles and associated DNA‐polymerase activity in saliva of chronic hepatitis B carriers
Authors: Macaya Gabriel , Visoná Kirsten A. , Villarejos Víactor M.
Published: March 2007

To investigate further the problem of salivary transmission of type B hepatitis, salivas free of blood contamination from three HBsAg‐positive carriers with chronic active hepatitis were examined by CsCl equilibrium density gradients and electron microscopy (EM). In the CsCl gradient HBsAg of whole salivas distributed in a band centered at 1.19 gm/cm3 with a clearly defined shoulder at 1.24 gm/cm3; the HBsAg was found mainly in the mucous component. On EM examination, fractions from the 1.19 gm/cm3 peak contained spherical HBsAg particles of 22+mn3 nm diameter, whereas in the 1.24 gm/cm3 shoulder Dane particles 43 nm in diameter with 28 nm cores were found. Specific hepatitis B virus associated DNA‐polymerase activity also was found in the 1.24 gm/cm3 shoulder where the Dane particles occurred and was absent from the saliva of healthy controls. When salivas were incubated for three hours at 37°C the total amount of HBsAg diminished and the 1.24 gm/cm3 shoulder disappeared, probably as a result of endogenous degradation of the Dane particles and the free HBsAg. These findings clearly indicate that the hepatitis B viral particle is present in the saliva of chronic HBsAg carriers with active disease and further confirm that saliva is an important vehicle of infection.
Antiviral activity of colostrum and serum immunoglobulins a and g
Authors: Palmer E.L. , Gary G.W. , Black R. , Martin M.L.
Published: March 2007

Enteric virus‐specific IgA and IgG present in paired human sera and colostrums were measured by the enzyme‐linked immunosorbent assay (ELISA). Virus‐specific IgA was present in all colostrums, but virus‐specific IgG could not be detected. The reverse was true when sera were assayed. Most of these colostrums also neutralized either polio virus or reovirus, as did IgA, which was separated from a pool of colostrums by exclusion chromatography. No correlation could be made between levels of neutralizing and ELISA antibody titers in colostrums.
Oyster‐Associated gastroenteritis in australia: the detection of norwalk virus and its antibody by immune electron microscopy and radioimmunoassay
Authors: Grohmann G. S. , Greenberg H. B. , Welch Bettina M. , Murphy A. M.
Published: March 2007

Following widespread outbreaks of oyster‐associated gastroenteritis in Australia during 1978 in which Norwalk virus was implicated as the causative agent, collaborative studies were undertaken between laboratories in Australia and the United States to confirm the etiology. Immune electron microscopy (IEM) techniques were used in Australia and radioimmunoassay (RIA) methods in the United States. Norwalk virus was detected by IEM in seven of 15 faecal samples, and four were positive by RIA. A much better correlation was found with antibody determinations. Both methods demonstrated significant increases in antibody to Norwalk virus in 22 of 30 sets (73%) of “acute” and “convalescent” sera, confirming that Norwalk virus was responsible for the majority of cases.

It is significant that the RIA serology was determined using Norwalk antigen originating in the United States and the IEM serology was determined using 27–30‐nm particles originating in Australia.
Humoral immune response to virions and dense bodies of human cytomegalovirus determined by enzyme immunofluorescence assay
Authors: And Bagher Forghani , Schmidt Nathalie J.
Published: March 2007

An enzyme immunoassay utilizing a fluorogenic substrate (EIFA) was employed to compare humoral antibody responses to purified virion and dense body antigens in human cytomegalovirus (CMV) infections. Results of these antibody assays were also compared to those obtained by plaque reduction neutralization and complement fixation (CF). In adults with CMV infections diagnosed by a significant rise in CF antibody titer, there was also a vigorous antibody response to the virion and dense body antigens. There was better correlation between antibody levels to the virion and dense body antigens than between antibody levels detected by other combinations of tests. Antibody responses to the virion and dense body antigens were similar, but not identical, suggesting that although the two structures share major antigens, they may also possess unique antigens to which the host can mount independent antibody responses. All individuals with CMV neutralizing antibody also had virion and dense body antibodies, but there was poor correlation between neutralization titers and those detected by EIFA, perhaps reflecting greater strain specificity of the neutralization test. Some individuals with serological diagnoses of Mycoplasma pneumoniae or influenza A infection showed fourfold or greater antibody titer rises to CMV in two or more assays, suggesting reactivation of latent CMV in these infections.
Preparation of a highly purified vaccine against tick‐borne encephalitis by continuous flow zonal ultracentrifugation
Authors: Heinz Franz X. , Kunz Christian , Fauma Hans
Published: March 2007

By the use of continuous‐flow zonal Ultracentrifugation, a highly purified, inactivated tick‐borne encephalitis (TBE) virus vaccine was prepared that contained almost exclusively TBE virus particles. The purity of this preparation was 90‐ to 100‐fold higher than that of a previously used partially purified vaccine. A mean protein content of 3–5 m̈g/ml in the final vaccine represents 20 mouse protective doses50 in a mouse challenge potency test. This high degree of purity resulted in a strong reduction of side reactions observed with the less purified vaccine.
Optimized enzyme‐linked immunosorbent assay for detection of human and bovine rotavirus in stools: Comparison with electron‐microscopy, immunoelectro‐osmophoresis, and fluorescent antibody techniques
Authors: Grauballe P. C. , Vestergaard B. F. , Meyling A. , Genner J.
Published: March 2007

Detection of human and bovine rotavirus in stools is described using a double‐antibody sandwich enzyme‐linked immunosorbent assay (ELISA) with polystyrene microtest plates as solid phase, immunoglobulin fraction of rabbit antiserum to rotavirus (human) as catching antibody, and the same reagent labelled with horseradish peroxidase as conjugate. The ELISA has been optimized with regard to simplicity, rapidity, sensitivity, and specificity. In a comparative study, stool specimens from 81 infants and children and 92 neonatal calves with diarrhoea were tested for rotavirus by ELISA, electron microscopy (EM), immunoelectro‐osmorphoresis (IEOP), and fluorescent antibody technique (FA). The relative sensitivity of the different assays for human and bovine rotavirus was: EM 68%, 76%; IEOP 80%, 76%; FA not determined, 85%; and ELISA 86%, 98%, respectively. Less than 1 ng of purified human rotavirus could be detected in ELISA, whereas 100 ng was the minimal amount detected by IEOP. It is concluded that the developed ELISA is a simple, rapid, reliable, and sensitive method for the diagnosis of human and bovine rotavirus infections.