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Overlapping fragments of the bovine herpesvirus-1 (BHV-1) glycoprotein (gD) ORF were expressed as trpE-gD fusion proteins in Escherichia coli to map linear neutralizing epitopes defined by BHV-1-specific MAbs. The MAbs 3402 and R54 reacted with the expressed fragments on Western blots that located the epitopes between the amino acids 52-126 and 165-216, respectively, of gD. Bovine covalescent sera with high neutralizing antibody titers against BHV-1 reacted with these bacterially expressed proteins containing both of the epitopes. Alignment of these sequences from BHV-1 with the corresponding region of the BHV-5 gD ORF sequences (reported here) identified several amino acid mismatches. Since the MAbs 3402 and R54 neutralize the BHV-1 and not BHV-5, it was presumed that these were important amino acids in defining the epitope. To further localize the neutralizing epitopes, synthetic peptides corresponding to these regions in the BHV-1 gD ORF were tested for their capacity to block monoclonal antibody neutralization of BHV-1 infectivity. The peptides encompassing amino acids 92-106 (3402 epitope) and amino acids 202-213 (R54 epitope) of the BHV-1 gD competed with BHV-1 for the binding by MAbs 3402 and R54, respectively, in a dose-dependent manner. Antisera produced in rabbits to these peptides conjugated to a carrier reacted strongly with a 30-kDa protein by Western blotting and had neutralizing antibody titers against BHV-1.

Reciprocal competition ELISA (rcELISA) was conducted to map monoclonal antibodies (mAbs) reactive with gI, gIII and gIV glycoproteins of bovine herpesvirus type 1 (BHV-1) into epitope groups. mAbs to glycoproteins gI and gIV were divided into six epitope groups each, while gIII mAbs had been previously divided into four areas. mAbs were chosen from each epitope group to compete in cELISA wih bovine sera collected during a typical regimen of vaccination and transportation from farm to auction to feedlot. The immunodominant epitopes were identified for each BHV-1 glycoprotein. With glycoprotein gI, three epitopes defined by mAbs 1F10, D9 and 4807 were the most dominant; with glycoprotein gIII epitopes defined by mAbs G2 and 1507, and with glycoprotein gIV epitopes defined by mAbs 1102, 1106, 3C1, 3402 and 3E7 showed the maximum responses. The overall cELISA responses to each glycoprotein among two vaccination groups were also compared and it was shown that cELISA responses were significantly higher for each glycoprotein in calves receiving two vaccinations, one on the farm of origin and one at auction, than in calves receiving only one vaccination at auction.

A type of bovine herpesvirus, BHV-1.3, causes encephalitis in calves, whereas BHV-1.1 causes respiratory disease. Three colostrum-deprived calves and two colostrum-fed calves were inoculated with BHV-1.3 by intranasal aerosolization. Two colostrum-deprived calves were inoculated with BHV-1.1 by intranasal aerosolization. BHV-1.3-inoculated calves demonstrated severe encephalitis with minimal respiratory lesions, and BHV-1.1-inoculated calves demonstrated severe respiratory lesions and no clinical signs of neurologic disease. Calves fed colostrum that contained virus neutralizing antibodies were protected against neurologic disease. Colostrum-fed BHV-1.3-inoculated calves did not develop disease although they did become infected; virus was shed in respiratory secretions for 10-13 days postinoculation, similar to infected colostrum-deprived calves. BHV-1.3 was reactivated from a latent state from one colostrum-fed calf after administration of dexamethasone 60 days postinoculation. Histopathologic examination of the three colostrum-deprived BHV-1.3-inoculated calves revealed severe lesions of encephalitis. One of the two BHV-1.1-inoculated calves had one focal lesion of encephalitis. Virus was isolated from brain tissue of colostrum-deprived BHV-1.3-inoculated calves and from one BHV-1.1-inoculated calf. Immunohistochemical staining for BHV-1 antigen was observed in neurons from the colostrum-deprived BHV-1.3-inoculated calves.

Seven mouse hybridoma cell lines producing monoclonal antibodies (MAb) against an encephalitogenic strain of bovine herpesvirus type 1.3 (BHV-1.3) were established. The clones producing MAb were selected to be specific for BHV-1.3 by enzyme-linked immunosorbent assay. Only L1B neutralized virus without complement. With the addition of complement, five of the MAb neutralized BHV-1.3 but not the respiratory strain BHV-1.1. The anti-BHV-1.3-specific MAb Q10B, L6G, and L1B precipitated glycoproteins from BHV-1.3 that were analogous to the gI, GIII, and gIV glycoproteins of BHV-1.1, respectively. The other four MAb precipitated unknown proteins. None of the anti-BHV-1.3 MAb precipitated BHV-1.1 glycoproteins. The majority of the anti-BHV-1.3 MAb did not react with BHV-1.1 by immunoblotting, but O7E (unknown protein pattern by radioimmunoprecipitation) was reactive with five proteins (M(r)s of 33,000, 43,000, 70,000, 141,000, and 190,000) of BHV-1.3 and with a different pattern of proteins of BHV-1.1 (M(r)s of 30,000, 38,000, 83,000 and 144,000). Two of the MAb, L6G and O7E, conjugated with peroxidase were found to be useful for detecting BHV-1.3 antigen by immunochemistry in Formalin-fixed brain tissue from experimentally infected calves.

An immunosuppressive factor produced by an esophageal squamous carcinoma cell line mediates profound irreversible suppression of in vitro proliferative responses of lymphoid cells. Exposure of activated normal PBL to the immune suppressive factor (ISF) resulted in the induction of an irreversible anergic state with apoptosis evident in 20% of those cells. Flow cytometric cell cycle analysis of mitogenically stimulated normal lymphocytes exposed to the ISF revealed that despite exhibiting full activation status (IL-2 production, IL-2R, and transferrin receptor expression) PBL were arrested at the G1/S interphase of the cell cycle. Transformed lymphoid cell lines, NSO and JURKAT, displayed morphologies characteristic of apoptosis within 24 h of exposure to the ISF. Flow cytometric cell cycle analysis of the JURKAT cells incubated with ISF revealed that > 90% of these cells had undergone apoptosis within 24 h. Agarose gel electrophoresis of DNA extracted from the ISF-treated lymphoid cells resolved a DNA fragmentation pattern characteristic of apoptosis in both the NSO cells and to a lesser extent in the activated PBL exposed to the ISF but not in control cells. In JURKAT cells stimulated with anti-CD3 antibodies, Ca2+ mobilization was markedly enhanced in those cells exposed to ISF. Also, ISF independently induced a calcium flux in JURKAT cells. Induction of programmed cell death by ISF may account for the in vivo immune suppression local to the tumor site in squamous carcinoma of the esophagus.

Differences in the antigenic structure of the major glycoproteins, gI, gIII and gIV, of bovine herpesvirus type 1.1 (BHV1.1) and the neurovirulent BHV1.3 were demonstrated with a panel of monoclonal antibodies (MAbs) prepared against the BHV1.1 glycoproteins. Glycoprotein gIII of BHV1.3 was the most dissimilar, reacting with only four of 15 gIII-specific MAbs. Glycoproteins gI and gIV of BHV1.3 reacted with eight of 11 and eight of 12 specific MAbs, respectively. Monospecific bovine antisera to the two viruses supported findings from the MAb analysis in that gI and gIV glycoproteins were cross-recognized, but gIII was not. Virus-neutralizing MAbs reactive to each glycoprotein and which reacted with both viruses also neutralized both viruses. Previously undescribed glycoproteins which were antigenically related to the intact gIII glycoproteins, but of reduced sizes and lacking at least one gIII epitope, were found for both viruses. Tunicamycin inhibition experiments and immunoprecipitation data suggested that these proteins were intracellular degradation products. Comparisons of the peptide footprints of the glycoproteins from the two viruses using protease V8 digestion after immunoprecipitation with cross-reactive MAbs revealed distinctive footprint patterns for the respective glycoproteins.

Risk to health during adolescence is more likely due to behaviour than disease. Problems arise because behaviour is distally removed from its consequences and negative reinforcement is not contiguous with it. The paper reviews recent research and the problems faced by health workers in devising educational programmes when behaviour, in the cultural value system of adolescents, is present-oriented and outcomes are future-oriented. The challenge to the behavioural scientist is to develop health education programmes which have personal meaning to adolescents. Working against this aim are such processes as adolescent taboos, locus of control, the personal fable, risk behaviours and social-cognitive immaturity. Suggestions are made for research scientists to devise and evaluate programmes to change the operational liberalism of adolescent behaviour by installing more ideologically conservative attitudes in the hope that behavioural change will follow.

An indirect immunoperoxidase (IP) procedure using the avidin-biotin-peroxidase complex detection technique was developed to detect viral equine herpesvirus-1 (EHV-1) antigen in formalin-fixed paraffin-embedded tissues from aborted equine fetuses. The procedure was applied to liver, lung, and other tissues from 20 cases of confirmed or suspected EHV-1-induced abortions. Specific staining was observed in tissue sections from EHV-1-infected fetuses. Positive IP staining was present in tissues of 7 cases that were also positive by fluorescent antibody (FA) and virus isolation (VI) and that had typical histologic lesions. There was no IP staining in 7 cases that had no histologic lesions and negative FA and VI results. Five cases had typical histologic lesions and positive results in only 1 laboratory test; 3 were positive by VI and 2 by FA. Liver of 1 case was positive by IP, but tissues were too autolytic for other tests to be evaluated.

Serologic evidence of infection by Toxoplasma gondii, feline leukemia virus, feline coronaviruses, or feline immunodeficiency virus (FIV) is commonly found in cats with uveitis. Serum samples from 124 cats with uveitis were assayed by use of ELISA for the detection of T gondii-specific immunoglobulin M (IgM), IgG, and circulating antigens (Ag), as well as an ELISA for feline leukemia virus Ag, an ELISA for antibodies to FIV, and an indirect fluorescent antibody assay for antibodies to feline coronaviruses. Serologic evidence of infection by 1 or more of the infectious agents was detected in 83.1% of the samples. Serologic evidence of T gondii infection, defined as the detection of T gondii-specific IgM, IgG, or Ag in serum, was found in 74.2% of the samples. The seroprevalence of T gondii infection was significantly greater in cats with uveitis than in healthy cats from a similar geographic area. Serum samples from cats with serologic evidence of both T gondii and FIV infections were more likely to contain T gondii-specific IgM without IgG than samples from cats with serologic evidence of T gondii infection alone. Cats with serologic evidence of FIV and T gondii coinfection had a higher T gondii-specific IgM titer geometric mean and a lower T gondii-specific IgG titer geometric mean than did cats with serologic evidence of T gondii infection alone. Serologic evaluation for T gondii infection should include assays that detect IgM, IgG, and Ag, particularly in cats coinfected with FIV.

The abomasa of 1,949 slaughtered feedlot cattle, 45 necropsied feedlot cattle that died 2 to 45 days after arrival, and 45 necropsied pastured cattle were opened and examined. Of these organs, 484, 1, and none, respectively, contained erosions. The slaughtered cattle were fattened at 3 locations: 1,305 with 430 eroded abomasa were fed a ration of corn in northeastern Colorado; 144 cattle with 4 affected abomasa fed a ration of milo in south-central Arizona; and 500 cattle with 50 affected abomasa fed a ration of milo and corn in northwestern Texas. The red-brown lesions developed late during the second semester of fattening and were located mostly on fundic folds. Those on fold edges were linear and were 2 to 15 cm long, whereas those on fold sides were punctate and were 2 to 15 mm in diameter. Normal fold edges contained fewer goblet cells and less surface mucus than did fold sides. Eroded folds had disruption of surface epithelium, damage to endothelial cells, and dilated, thrombosed, congested, and ruptured capillaries. Mean pH values of 16 normal and 17 eroded abomasa were 4.7 and 3.9, respectively. Necrosis of all tissue toward the mucosal surface of erosions was extensive. The cause of gastric erosion in cattle is not known.

Endemic cretinism is the most severe manifestation of dietary iodine deficiency. Two forms of the syndrome are traditionally described: neurological and myxoedematous. Although this classification highlights the important neurological sequelae of the disorder it implies that myxoedematous cretins have an alternative mechanism. Further, the nature of the neurological deficit associated with both types of endemic cretinism has received scant attention in recent times considering that it remains a common disorder in many parts of the world. The nature and extent of the neurological deficit found in endemic cretinism was investigated in 104 cretins from a predominantly myxoedematous endemia in western China and in 35 cretins from central Java, Indonesia, a predominantly neurological endemia. We found a similar pattern of neurological involvement in nearly all cretins from both endemias, regardless of type (myxoedematous or neurological), and of current thyroid function. Hallmarks of the neurological features included mental retardation, pyramidal signs in a proximal distribution and extrapyramidal signs. Many patients exhibited a characteristic gait. This probably reflected pyramidal and extrapyramidal dysfunction, although joint laxity and deformity were important contributing factors. Other frequently encountered clinical features were squint, deafness, and primitive reflexes. Cerebral computerized tomography (CT) revealed basal ganglia calcification in 15 of 50 subjects. The presence of basal ganglia calcification was confined to cretins with severe hypothyroidism. Otherwise, cerebral CT scanning demonstrated only minor abnormalities which did not contribute to the localization of the clinical deficits. We conclude that the same neurological disorder is present in both types of endemic cretinism reflecting a diffuse insult to the developing fetal nervous system. These clinical findings support the concept of maternal and fetal hypothyroxinaemia, arising from severe iodine deficiency, as the primary pathophysiological event in endemic cretinism. Differences between the two types of cretinism may be explained by continuing postnatal thyroid hormone deficiency in the myxoedematous type, which results in impaired growth, skeletal retardation and sexual immaturity.

A ribonucleic acid (RNA) hybridization assay to identify cattle infected by bovine viral diarrhea virus (BVDV) is described. The RNA probe was derived from the coding region at the 3' end of the genome of the NADL strain of BVDV. Total RNA from infected cell cultures or peripheral blood leukocytes from suspect animals was extracted and applied to nylon membranes with a slot blot apparatus. Peripheral blood leukocytes were tested concurrently for BVDV by virus isolation. The results of hybridization and virus isolation were in agreement for 92% of the cases. When compared with virus isolation, hybridization had a sensitivity of detection of 59.5% and a specificity of 95%. Cross-reactivity to RNA extracts of border disease virus-infected cells was noted. No cross-reactivity was detected to other common bovine viruses (bovine herpesvirus-1, bovine respiratory syncytial virus, parainfluenza-3 virus, and bluetongue virus), to viruses classified in related families (equine arteritis virus and Venezuelan equine encephalitis virus), or to viruses having similar genomic organization (dengue virus type 2 and Japanese encephalitis virus).

Approximately 10 of 100 young heifers that had recently delivered their first calf--members of a large Colorado dairy herd--had a syndrome of swollen teats and distal portions of the hind limbs, prefemoral lymphadenopathy, transient fever, rough coat, decreased milk production, and subsequent weight loss and reproductive inefficiency. Acute clinical signs of disease were associated with large numbers of Eperythrozoon wenyonii seen on blood smears, and resolution of signs correlated with reduction or disappearance of the parasite. Other known causes of peripheral edema could not be documented. The parasite was transmitted to 4 of 7 nonlactating dairy cows destined to be culled and a splenectomized calf via IV inoculation of blood from parasitemic heifers, but clinical signs of infection were not induced.

Experiments were undertaken to determine the tissue distribution of Escherichia coli and a coliphage after contamination of the common mussel (Mytilus edulis). Mussels were contaminated with high levels of feces-associated E. coli and a 22-nm icosahedral coliphage over a 2-day period in a flowing-seawater facility. After contamination, individual tissues were carefully dissected and assayed for E. coli and the coliphage. Contaminated mussels were also analyzed to determine the tissue distribution of the contaminants after 24- and 48-h depuration periods. The majority of each contaminant was located in the digestive tract (94 and 89% of E. coli and coliphage, respectively). Decreasing concentrations were found in the gills and labial palps, foot and muscles, mantle lobes, and hemolymph. Our results indicate that contamination above levels in water occurred only in the digestive tract. Contaminated mussels were depurated in a commercial-scale recirculating UV depuration system over a 48-h period. The percent reductions of E. coli occurred in the following order: digestive tract, hemolymph, foot and muscles, mantle lobes, and gills and labial palps. The percent reductions of the coliphage were different, occurring in the following order: hemolymph, foot and muscles, gills and labial palps, mantle lobes, and digestive tract. Our results clearly demonstrate that E. coli and the coliphage are differentially eliminated from the digestive tract. The two microorganisms are eliminated at similar rates from the remaining tissues. Our results also clearly show that the most significant coliphage retention after depuration for 48 h is in the digestive tract. Thus, conventional depuration practices are inappropriate for efficient virus elimination from mussels.

Studies were undertaken to determine the effect of temperature, salinity, and food availability on the efficiencies of elimination of Escherichia coli and a 22-nm icosahedral coliphage from experimentally contaminated mussels. Test temperatures (5.5, 10, 16.5°C) and salinities (18.2, 28.6 ppt) reflected normal seasonal fluctuations during routine commercial depuration. The initial E. coli levels were reduced by >99% within 52 at all temperatures. In contrast, efficient coliphage elimination occurred at 16.5°C only. The initial E. coli levels were reduced by >99% at both salinities, while coliphage elimination was relatively inefficient under similar conditions. In unfiltered seawater, the addition or omission of food, in the form of Tetraselmis suecica , had no appreciable effect on either E. coli or coliphage elimination from mussels. In filter-clarified seawater, E. coli elimination was more efficient and coliphage elimination was considerably enhanced when food was added. In the absence of food, coliphage elimination was very inefficient. The results of these studies indicate that bacterial elimination from mussels during depuration is efficient through the range of parameters used. In contrast, coliphage elimination was generally inefficient throughout the study, suggesting that depuration, as currently practiced, cannot be relied upon to render mussels completely free of viral contamination. These studies emphasize that successful bacterial depuration does not reflect viral elimination and therefore, bacterial standards for efficient depuration of viruses are unreliable.

An erythroleukemia cell clone, 7C, which failed to produce reverse transcriptase-containing virions or infectious virus, was found to produce noninfectious virus particles by gradient banding of [3H]leucine- and [3H]uridine-labeled virions. The RNA from the 7C virus was shown to consist of the normal 70S size component, which converted to 35S upon heat denaturation. In contrast, the 7C virion proteins showed multiple defects. Analysis of the virion proteins by gel electrophoresis demonstrated that the pr65 gag precursor was incorporated into the 7C virus and that the processing of this precursor was severely diminished. Polymerase proteins pr180gag-pol and pr120pol were also detected in virions, and a third possible polymerase protein, p70, was reduced in size compared to its normal counterpart, p80. Incorporation of the viral gp70 glycoprotein into particles was also reduced 10-fold, despite synthesis and incorporation of gp70 into the 7C cell membrane in normal amounts. Pulse-chase analysis of the synthesis of the viral gag and env proteins in 7C cells showed greatly reduced amounts of pr180gag-pol, pr65gag, p80gag, and p42gag, whereas pr90env, gp70, and spleen focus-forming virus-specific gp55 were synthesized and processed normally. These results suggested that at least one defect in 7C virus was impaired cleavage of gag or pol proteins or both, most likely due to a lack of the appropriate viral protease, and that this lack of cleavage might affect incorporation of gp70 into virus particles.

Treatment of influenza virus and its purified hemagglutining with glycosidases from Diplococcus pneumoniae, which included beta-galactosidase, beta-N-acetylglucosminidase, and endoglycosidase D, released amino and neutral sugars from the virus and these as well as large oligosaccharides from the purified hemagglutinin. The released glucosamine-containing oligosaccharides were of one discrete size. Large oligosaccharides not removed by the glycosidases were found on the virus as well as the hemagglutinin. Some oligosaccharides on the virus were inaccessible to the enzymes, since they could be removed only from the purified hemagglutinin. Approximately 50% of the hemagglutinin carbohydrates could be removed without effect on hemagglutinating activity. Similarly, removal of 20 to 25% of the carbohydrates from intact virus particles did not alter infectivity.

Forty-five cases of bovine abortion were examined using in situ hybridization (ISH) with a biotinylated DNA probe specific for bovine herpesvirus-1 (BHV-1). Of the 45 cases, 16 were diagnosed as due to BHV-1, 15 were determined to be due to other causes, and 14 were of undetermined etiology. Direct comparisons between ISH and an immunoperoxidase (IP) test specific for BHV-1 were performed on formalin-fixed, paraffin-embedded tissue sections of lung, liver, kidney, spleen, thymus, and placenta; fluorescent antibody tests for BHV-1 and virus isolation were performed on fresh lung and liver. In comparison to these routine BHV-1 detection techniques, ISH had an overall sensitivity of 88.2% and a specificity of 89.3% in detecting BHV-1 in aborted fetuses. Immunoperoxidase was more sensitive than ISH with tissue sections from lung (87.5% vs. 69%), liver (92% vs. 17%), spleen, and placenta; results of the tests on tissue sections from kidney were concordant. Liver sections presented special problems in that nonspecific reactions were frequently observed with hybridization. With thymus sections, the rate of detection was higher by hybridization than by IP, but the specificity of some of these reactions could not be confirmed.

Both dating behavior and peer expectations, in 317 adolescents, were examined during the first date, after several dates, when going steady, and when some commitment to marriage was undertaken. The results showed an initial tendency for the behavior of males to be more intimate than that of females. Female behavior approached that of males as the commitment in the affectional relationship increased. Generally, both males and females conformed to their peer expectations for less intimate behaviors but not for the deeper forms of sexual embrace, where they imagined their peers to be more experienced.

A total of 457 nasal swab specimens from cases of respiratory disease in 2 feed lots were evaluated for the detection of bovine herpesvirus Type 1 (BHV-1) by ELISA. Thirty-three were found to be positive for BHV-1 by the recovery of infectious virus and 21 of these were positive by ELISA, yielding a sensitivity of 64%. Fifteen other virus isolations were made and included bovine viral diarrhea viruses, rhinoviruses and parainfluenza Type 3 viruses; none of these cases were positive with the BHV-1 ELISA. Specificity of the ELISA was 100%. Eighty percent of the specimens with BHV-1 titers greater than 10(5) TCID50 were detected by ELISA; the median amount of virus in positive specimens that were detected by ELISA was 7 X 10(5) TCID50 and the median amount of virus in specimens not detected was 1.5 X 10(4) TCID50. BHV-1 infection was most frequently diagnosed in feedlot cattle that had been in the feedlot for 40-80 days. Approximately half of the infected cattle were carrying virus-neutralizing antibodies in their serum.

An enzyme-linked immunosorbent assay (ELISA) for the detection of cattle antibodies to bovine herpesvirus type 1 was developed on the basis of competition between serum antibody and a virus-neutralizing mouse monoclonal antibody. The assay showed improved sensitivity over the virus neutralization (VN) test and over an enhanced VN test in which incubation of antibody-virus mixtures was carried out for 24 h. With the ELISA, antibodies in sera from experimentally infected cattle were detected earlier after infection and showed more rapid increases in levels. A comparison of the ELISA with the VN tests by using a set of 85 field sera with low levels of antibodies demonstrated that the ELISA was the most sensitive test, detecting 10 positive serum samples that were negative by the VN tests. The ELISA was inexpensive, rapid, and highly reproducible and showed a significant improvement in sensitivity over VN tests.

Video-image representations of body size were measured on a television monitor that was modified to give a display continuum that ranged from 50% under- to 50% over-estimation of objective size. Sixty-eight females who were undergoing weight reduction counseling were measured before and after treatment. All Ss judged themselves to be significantly more obese than they actually were, with a tendency for errors to be greatest among the more obese Ss. After therapy, more realistic estimates of their physiques ensued. A drop-out rate of 23% was recorded; the drop-outs saw themselves as significantly more obese than those who graduated from the program.

Thirty-three cattle with fatal respiratory tract disease were examined for gross and histologic lesions and for the presence of viral and bacterial agents in the lungs. Fifteen cattle had lesions characteristic of atypical interstitial pneumonia (AIP), and 18 had other respiratory tract diseases, including infectious bovine rhinotracheitis, shipping fever pneumonia, bronchopneumonia, pulmonary abscess, and edema of the trachea. Gross necropsy findings in the cattle with AIP were uncollapsed and emphysematous lungs; histopathologic findings included interstitial edema, thickening of alveolar walls, hyaline membrane formation, and hyperplasia of type-II pneumonocytes. The infective agents found in the lungs of the 33 cattle included bovine respiratory syncytial virus, bovine herpesvirus type 1, Pasteurella sp, mycoplasmas, and Corynebacterium pyogenes. Bovine respiratory syncytial virus was detected by use of immunofluorescence and immunoperoxidase on lung tissue sections; bovine herpesvirus type 1 was detected by these techniques and by isolation of the virus. Bovine respiratory syncytial virus was significantly (P = 0.01) associated with lesions of AIP (11 of 15), compared with those of other respiratory tract diseases (5 of 18).

Intelligence was measured by means of the Hiskey-Nebraska Test of Learning Aptitude or the Griffiths Mental Development Scales in a sample of 369 patients from iodine-deficient rural villages (Baihuyao), iodine-sufficient rural villages (Huanglo) and urban populations to test for the damaging effects of iodine deficiency on the development of the nervous system in the presumed healthy section of a community. In urban school-children who were aged seven to 14 years (n = 78), a normal range of measured intelligence was found (mean +/- SD intelligence-quotient score, 107.0 +/- 18.3). By comparison, intelligence-quotient scores were lower in all rural cohorts (a rural suppression effect) but the distribution of intelligence-quotient scores showed a further shift to the left in the iodine-deficient township. In Baihuyaon villagers who were aged 30-35 years (n = 50), who were born during the period of severe iodine deficiency, 72% of villagers had an intelligence-quotient score of less than 70 compared with 41% (P less than 0.05) of villagers who were aged 28-35 years from Huanglo, a rural iodine-sufficient control group (n = 49). Although measured intelligence was higher in Baihuyaon children whose mothers had received iodized salt - 44% of seven- to 14-year-old children had intelligence-quotient scores of less than 70 (n = 141)--it remained significantly depressed compared with rural (n = 51) and urban (n = 78) control subjects (18% and 4%, respectively). These findings were consistent with other parameters (that is, the persistently-high rate of goitre), which indicated that the salt-iodization programme was inadequate. In the iodine-deficient village, lower intelligence-quotient scores showed a relationship with the detection by audiometry of nerve deafness and with the presence of abnormal neurological signs. The latter included spasticity and pyramidal signs which were of a similar pattern to the neurological deficits that have been demonstrated in overt neurological cretins. We conclude that iodine deficiency imposes a further suppressive effect on the intellectual performance of rural inhabitants, and results in a shift of the entire population distribution of cognitive skills to a lower level.

Simian virus 40 gene A has previously been shown to promote the replication of viral DNA and the transcription of late viral RNA in productive infection and to maintain the growth characteristics of some transformed cells. The present study examines the effect of the A function on proteins synthesized during productive and transforming infections. Under restrictive conditions, temperature-sensitive A mutants induce the overproduction of a 100,000-dalton protein both in productively infected monkey cells and in transformed rabbit cells. Immunoprecipitation of the induced protein with antisera, prepared against simian virus 40-induced tumors in hamsters, was used to identify the induced protein as tumor antigen. The same protein can be precipitated from extracts of cells infected by wild-type virus but not from uninfected cells. Furthermore, the mutant-induced protein is more rapidly degraded in vivo and is less tightly bound to intranuclear components than the protein induced by wild-type virus. The presence of the same virus-induced protein in infected cells from different species and the altered behavior of that protein in mutant infection strongly suggest that the protein is virus coded. Because the protein is large enough to account for the entire coding capacity in the early region of the simian virus 40 genome, the 100,000-dalton protein may well be the primary product of the only early gene identified by complementation studies, the A gene. If the 100,000-dalton protein that is overproduced in mutant infection is the A protein and the only early protein, then functional wild-type A protein must regulate its own synthesis in both productive and transforming infections.

Bovine viral diarrhea virus (BVDV) antigen was detected in cell culture with an indirect immunoperoxidase (IP) procedure using a specific monoclonal antibody, and an avidin-biotin-peroxidase complex. Cytopathic and noncytopathic strains of the virus showed similar patterns of IP staining until 3 days post-infection. At six days post-infection, intensity of staining decreased in cell cultures infected with noncytopathic virus, but not with cytopathic virus. The IP procedure detected BVDV antigen in cells used to isolate virus from tissues of aborted bovine fetuses and peripheral blood lymphocytes of adult cattle. Immunoperoxidase detected BVDV isolates from 10 of 44 cases of abortion of which seven of these were noncytopathic. Noncytopathic BVDV isolates from the peripheral blood lymphocytes of 7 of 65 animals were identified.

The Secord Homonym Test was used to elicit unconscious concerns about the body in one normal and two obese samples. The first sample of nonobese subjects was used to gather baseline data and also to examine the discriminant validity of the test as a measure of unconscious rather than conscious body concern. In a second sample of obese subjects a significantly greater concern with the body was observed. A third sample of obese subjects was tested before and after weight reduction counseling. Initial measures showed them to have similar levels of concern to the second sample; however, following treatment the levels matched those of the nonobese subjects.

Spleen cells from Rfv-3r/s mice with Friend virus-induced erythroleukemia were analyzed for expression of virus-induced proteins with monoclonal antiviral antibodies and conventional antisera. Leukemic spleen cells, 30-60 d after virus inoculation, expressed decreased amounts of ecotropic Friend murine leukemia helper virus gag- and env-encoded cell surface and intracellular proteins compared with leukemic cells tested 8-10 d after virus inoculation. In contrast, the spleen focus-forming virus-induced protein, gp55, was present on both leukemia cell populations. This difference appeared to be mediated by the humoral antibody response in Rfv-3r/s mice, which could recognize only ecotropic gag and env proteins, and not gp55. A new gp70 molecule cross-reactive with a recombinant Friend mink cell focus-inducing virus was found in large quantities on late leukemic cells. This protein appeared to be derived from a recombinant virus produced during the course of Friend virus infection. The appearance of this new gp70 suggests that recombinant viruses other than spleen focus-forming virus may play a role in Friend virus-induced erythroleukemia.

The virus specificity of antibodies against bovine herpes virus type 1 was determined with a radioimmunoprecipitation assay and serum collected from natural and experimentally induced infections. By using sequentially collected sera, the development of antibodies to 4 to 5 viral glycoproteins and 11 to 12 nonglycosylated proteins was followed for the first 50 days after infection. The major and most consistent responses in experimentally and naturally infected animals were to four glycoproteins with molecular weights of 102,000, 96,000, 69,000, and 55,000, as well as to a major virion 115,000-molecular-weight nonglycosylated protein. The four glycoproteins were all coprecipitated by a neutralizing monoclonal antibody and were probably involved as target antigens in virus neutralization. Another antigenically unrelated glycoprotein with a molecular weight of 82,000 and a nonglycosylated protein with a molecular weight of 91,000 were also precipitated, but the immune response to these two proteins was transient. Reactivity to gp82 was only weakly detected in serum from naturally infected animals. Contact control animals which did not contract a bovine herpes virus type 1 infection but were exposed to infected animals with signs of severe illness had antibodies which recognized gp102, gp96, gp69 and gp55 as well as p115. These antibodies were present in low amounts and, in contrast to infected animals, did not increase between acute and convalescent sampling.

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of bovine herpesvirus type 1 (BHV-1) antigens in nasal swab specimens collected from infected animals. Development of the ELISA involved screening and selection of BHV-1-specific monoclonal antibodies for their ability to capture BHV-1 antigens and for their stability and activity after conjugation to horseradish peroxidase. Forty combinations of capture-conjugate monoclonal antibody pairs were screened for detection of nanogram amounts of purified BHV-1 by using a double-antibody-sandwich ELISA in which antigen and conjugated antibody were simultaneously added to antibody-coated wells. Of the 40 monoclonal antibody pairs, 4 were analyzed further and 1 was selected for routine application to clinical specimens. Of 129 nasal swab specimens collected during the first 10 days after experimental infection with BHV-1, 66 were found to be positive by both virus isolation and ELISA and 34 were positive for infectious virus but negative by ELISA. One specimen was positive by ELISA but negative by virus isolation, and the remaining 28 specimens were negative by both tests. Quantitation of the virus-containing specimens showed that the ELISA had a lower detection limit of 10(3.5) median tissue culture infective doses. The ELISA was judged to be highly useful for diagnosis of BHV-1 infections, since all of the nasal swab specimens that were collected from 12 animals during the first 5 days of the infection, when the clinical signs were the most apparent, were positive.

Monoclonal antibodies were used to develop a double antibody enzyme-linked immunosorbent assay for the detection of canine parvovirus (CPV) antigen in fecal samples. The assay was specific for the hemagglutinating protein of CPV and detected as little as 1.5 ng of virus within a 15-minute incubation period. The use of monoclonal antibodies against 2 epitopes on the CPV antigen permitted the simultaneous addition of test sample and enzyme-conjugated antibody, thus considerably simplifying the manipulations required for the assay. Results were visually determined without special instrumentation. Clinical studies revealed greater than 95% correlation between enzyme-linked immunosorbent assay results and hemagglutination titers.

Electron microscopic examination of fecal specimens from adult dairy cows indicated seasonal coronavirus shedding. Fifty-two cows from a 300 cow herd were monitored for shedding of coronavirus. Approximately 50% to 60% of the cows monitored shed coronavirus during the winter months (November to March) of the first year of the study. During 2 subsequent years of monitoring the same cows, 20% to 30% of the cows shed coronavirus during the winter months. Virus shedding was not detected during the summer months (July to September). Half of the cows monitored were vaccinated with a modified-live rotavirus-coronavirus-Escherichia coli combination vaccine; however, vaccination did not influence seasonal shedding of coronavirus, as compared with shedding in the nonvaccinated cows. In nonvaccinated cows that calved in the winter months, the incidence of coronavirus shedding increased from 20% to 30% during the last 2 months of gestation to 65% to 70% at parturition. In vaccinated cows, the incidence of shedding did not increase at parturition.

The problems of adjustment during the midadolescent years were examined in 1298 privileged and underprivileged adolescents from the Sydney Metropolitan Area, Australia. Girls reported more problems than boys in all areas of adjustment and underprivileged adolescents more problems than the privileged group; however, a significant interaction effect was also found. Whether high-income or low-income group, the overwhelming number of problems were associated with educational adjustment and were interpreted as the reaction to pressures to achieve imposed on adolescents of this age.

A panel of monoclonal antibodies (MAbs) to BHV-1, specific for gI and gIII glycoproteins and for a nonglycosylated core protein p100, was used to examine the epitope specificity of the immune response to the virus in naturally exposed and experimentally infected cattle. Sera from experimentally infected calves were analyzed in a competition ELISA (C-ELISA). Antibody to an epitope on gIII appeared as early as 4 days post infection, although virus-neutralizing antibody did not appear until day 8 or later. Antibody production peaked at 13 to 18 days post infection but the level of antibody to each gIII epitope varied. Competition by sera from cattle naturally exposed to BHV-1 was analyzed as a function of the virus neutralization (VN) titer of these sera. Competition with most of the MAbs correlated linearly with neutralization titer. However, competition with 2 of the MAbs, one specific for gIII and one specific for gI, was maximal even at the lowest neutralization titer, and competition with another MAb, specific for a non-glycosylated core protein, p100, was negative. Selected sera from the naturally exposed group were also examined by radioimmunprecipitation, and were shown to react predominantly with gI, gIII and gIV glycoproteins and in a few shown to react predominantly with gI, gIII and gIV glycoproteins and in a few MAbs were determined, and it was found that neutralization was enhanced by certain combinations. A mutually exclusive relationship between neutralization enhancement and competition for binding by MAbs (as determined by reciprocal C-ELISA) indicated an epitope-specific, rather than antibody-specific, mechanism for neutralization enhancement.

Cell line Y57, obtained from a (C57BL/10 X A.BY)F1 mouse made leukemic with Friend virus complex (FV), spontaneously ceased FV production in vitro after 20 passages. By limiting dilution cloning, the incidence of non-virus-producing cells was found to increase with sequential passage. Nonproducer cells apparently arose spontaneously and were able to increase in number relative to producer cells because the growth rate of nonproducer cells was more rapid than the rate at which virus could reinfect them. Nonproducer clones all contained the defective spleen focus-forming virus in a latent but rescuable form but showed defects in helper virus env and gag gene product synthesis. Stable long-term virus-producing clones were obtained both by cloning at early in vitro passages or by cloning immediately after reinfection of nonproducer cells with FV.

The assumption that males approach dating from a pronounced psychobiological orientation while females approach it from a psychoaffectional orientation was questioned. The sex role adopted by the individual was considered to be as important a variable as biological sex. Male and female subjects from three age groups, 16-17 years, 19-20 years, and 24-25 years, completed questionnaires designed to measure their sex roles and dating orientations. All groups of males were found to approach the dating relationship from both a psychoaffectional and psychobiological orientation, while all groups of females approached it from a psychoaffectional orientation and showed an increase in psychobiological orientation with increasing age and increasing depth of relationship. Significant differences were also found in dating attitudes between male and female subjects adopting different sex roles. It was concluded that neither masculinity and femininity, nor psychobiological and psychoaffectional attitudes to dating, lie on single continua. Masculine and feminine sex roles as independent dimensions influence psychobiological and psychoaffectional orientations to dating which are in themselves independent dimensions and not ends of a single continuum.

Monoclonal antibodies were used to study neutralizing determinants on polypeptides of bovine herpesvirus 1. Two of three monoclonal antibodies which recognized nonoverlapping epitopes on a glycoprotein of 82,000 daltons were found to neutralize. A second group of monoclonal antibodies that individually precipitated five viral glycopolypeptides ranging in size from 102,000 to 55,000 daltons also neutralized. Two monoclonal antibodies which were the most efficient in neutralization recognized a non-glycosylated protein of 115,000 daltons which was the major polypeptide on the virus. A fourth group of monoclonal antibodies precipitated a non-glycosylated polypeptide of 91,000 daltons and several smaller polypeptides, but these antibodies demonstrated only limited neutralizing activity.

Previous developmental studies of visual shape judgements have failed to instruct subjects clearly as to the type of shape judgements required. Using the method of adjustment 180 subjects in five age groups, 6, 8, 10, 12 and 19 years, were instructed to judge either the real or projected shape of an elliptical standard slanted 39, 57 and 72 degrees from the fronto-parallel. No age changes were found in the tendency towards shape constancy. Objective shape judgements exhibited a significantly greater degree of constancy than projective judgements and there was no decline in constancy with increasing slant of the standard under objective instructions, in contrast to a marked decline under projective instructions. Female subjects consistently manifested a greater tendency towards shape constancy than males.

A large simian virus 40 (SV40)-specific protein can be efficiently immunoprecipitated from infected cell extracts with antisera obtained from hamsters bearing SV40-induced tumors. The protein has an apparent molecular weight of 88,000 to 100,000 with respect to markers with known molecular weights, but behaves anomalously on sodium dodecyl sulfate (SDS)-polyacrylamide gels. Cell lines infected by two different strains of SV40 synthesize immunoreactive proteins that differ slightly in mobility during SDS-polyacrylamide gel electrophoresis, evidence that the protein is coded for by the virus. These differences in protein size correlate with differences in the electrophoretic mobility of viral DNA fragments obtained by digestion with HindII and III restriction enzymes. The size of the viral capsid proteins VP2 and VP3 also varies with the strain of virus. dl-1001, a constructed deletion mutant that lacks part of the SV40A gene, directs the synthesis of a 33,000-dalton polypeptide that is not detected in cells infected with wild-type virus. The deletion fragment, like the larger protein, is phosphorylated. Maps of tryptic peptides from the 88,000- to 100,000-dalton protein and the 33,000-dalton fragment show common peptides and provide strong direct evidence that the proteins are products of the SV40 A gene. The deletion fragment reacts with antitumor sera and binds to double-stranded DNA in the presence of the complete A protein.

Clinical observation and collection of biological specimens from a large beef feedlot (approximately 30,0000 animals) were used to evaluate 6 approaches for validation of a disease reporting system. Data collected during a 12-month period were used to evaluate each approach. A subsample of disease cases reported to the National Animal Health Monitoring System (NAHMS) was compared with the clinical observations of the investigators. Although the agreement between clinical diagnosis by the NAHMS veterinarian and by feedlot health crews was high, the sensitivity and specificity of specific diagnoses varied from 100 to 18% and from 99 to 76%, respectively, which suggests that regular clinical observations by a veterinarian are needed to validate disease diagnoses reported to NAHMS by producers. Subsampling of a group of cattle by means of paired serologic determination of antibodies to infectious bovine rhinotracheitis virus, bovine viral diarrhea virus, bovine respiratory syncytial virus, and parainfluenza-3 virus revealed a high serologic conversion rate to infectious bovine rhinotracheitis, and high levels of preexisting antibody to bovine respiratory syncytial and parainfluenza-3 viruses. It was concluded that the current method of data collection for Colorado feedlots provides an acceptable level of sensitivity and specificity for the program. However, disease events that are not of economic importance to the feedlot operator will be underestimated. If an objective of NAHMS is to develop a base line of animal health conditions, diagnosis of diseases by current methods will be satisfactory. Occasional validation through clinical observations by a veterinarian will suffice to monitor quality of collected data.(ABSTRACT TRUNCATED AT 250 WORDS)

A molecular hybridization technique using biotinylated DNA probes was used to detect bovine herpesvirus 1 (BHV-1) nucleic acid species immobilized on nitrocellulose. Seventeen recombinant plasmids containing HindIII restriction fragments of the BHV-1 genome were compared for their ability to detect immobilized BHV-1 DNA from purified virus and infected cells. One probe, pCB2, labeled by nick translation with either 3H or biotin, detected as little as 10 pg of viral DNA. In time course experiments, BHV-1 DNA could be detected by 2 h postinfection in 10(6) infected cells. BHV-1 DNA was detected in nasal swabs and exudate from experimentally infected cattle, even when specimens had been stored for over a year. In a retrospective study of a respiratory disease outbreak in a feedlot, hybridization was compared with virus isolation for diagnosis of BHV-1 infections. The sensitivity rate was 0.68 with virus isolation as the referent standard. Blot hybridization provides a novel approach with unique applications for the diagnosis of bovine herpesvirus infections.