Stress-induced sickness behaviors: an alternative hypothesis for responses during maternal separation.

During maternal separation, some primate and nonprimate species show a biphasic (active/passive) response. The second stage is characterized by reduced activity, a hunched body posture, and other behaviors. Traditionally, the second stage has been referred to as "despair" and is considered an animal model for human depression. Recent research in psychoneuroimmunology suggests an alternative hypothesis--that behaviors occurring during the second passive phase represent stress-induced "sickness behaviors." This perspective more readily accounts for findings in widely divergent species, does not require assumptions regarding the ability to express complex emotional states, is empirically testable, and aligns the separation model with recent hypotheses regarding the nature and ontogeny of depressive illness.

Glucocorticoids affect gonadotropin-releasing hormone immunoreactivity in musk shrew brain.

Multiple interactions between the hypothalamic-pituitary-adrenal and the hypothalamic-pituitary-gonadal systems exist. In this study, we asked if glucocorticoid administration affected gonadotropin-releasing hormone (GnRH) immunoreactivity. We found that musk shrews treated with dexamethasone (DEX), a synthetic glucocorticoid, had more GnRH-immunoreactive (ir) cells in the forebrain than did cortisol- or control-treated animals. The effects of DEX were noted rapidly, within 15 min, after administration. These effects were observed in the forebrain as a whole and also in specific subpopulations of GnRH-ir cells located in the medial septum/diagonal band and the hypothalamus.

A review of rapid prototyping (RP) techniques in the medical and biomedical sector.

The evolution of rapid prototyping (RP) technology is briefly discussed, and the application of RP technologies to the medical sector is reviewed. Although the use of RP technology has been slow arriving in the medical arena, the potential of the technique is seen to be widespread. Various uses of the technology within surgical planning, prosthesis development and bioengineering are discussed. Some possible drawbacks are noted in some applications, owing to the poor resolution of CT slice data in comparison with that available on RP machines, but overall, the methods are seen to be beneficial in all areas, with one early report suggesting large improvements in measurement and diagnostic accuracy as a result of using RP models.

A search for Ebola virus in animals in the Democratic Republic of the Congo and Cameroon: ecologic, virologic, and serologic surveys, 1979-1980. Ebola Virus Study Teams.

More than 30 years after the first outbreak of Marburg virus disease in Germany and Yugoslavia and 20 years after Ebola hemorrhagic fever first occurred in central Africa, the natural history of filoviruses remains unknown. In 1979 and 1980, animals in the Democratic Republic of the Congo and Cameroon were collected during the dry season near the site of the 1976 Ebola hemorrhagic fever epidemic. The study objectives were to identify local animals and search for evidence of Ebola virus in their tissues. A total of 1664 animals representing 117 species was collected, including >400 bats and 500 rodents. Vero and CV-1 cells and IFA and RIA were used for virus and antibody detection, respectively. No evidence of Ebola virus infection was found. This study was limited in time and animal collections and excluded insects and plants. Long-term, prospective, multidisciplinary comparative studies will yield more information than will repeat short forays on the ecology of filoviruses.

A novel 5-lipoxygenase inhibitor prevents gallstone formation in a lithogenic prairie dog model.

Gallstone formation is dependent on biliary cholesterol supersaturation, the pronucleating effects of gallbladder mucin, and inflammation. We evaluated the effect of aspirin (ASA) and a 5-Lipoxygenase inhibitor (FLAPI) on cholesterol precipitation and leukotriene levels in an animal model of cholesterol gallstone formation. Male prairie dogs were divided into four dietary groups: normal chow controls, 1.2 per cent cholesterol (XOL), 1.2 per cent cholesterol plus ASA (XOL + ASA, 100 mg/kg/d), and cholesterol plus FLAPI (XOL + FLAPI, 100 mg/kg/12h). At 3 weeks the subjects were anesthetized, cholecystectomy performed, and the common duct cannulated for bile sampling. Cholesterol precipitation, lithogenic indices, and leukotriene content were analyzed. The group XOL + FLAPI did not form cholesterol crystals, whereas the group XOL + ASA did (P < 0.05, Fisher's exact test). All cholesterol-fed groups had significantly increased lithogenic indices when compared to controls. The XOL + FLAPI group showed a significant and paradoxical increase in LTB4 compared to the other groups (P < 0.05, ANOVA, Fisher's PLSD). This study has shown a significant decrease in the rate of cholesterol stone formation through the use of a novel leukotriene inhibitor at high doses, despite a high cholesterol diet.

The crystal structure of a human nucleoside diphosphate kinase, NM23-H2.

The 2.8 A resolution X-ray structure of NM23-H2 has been determined by molecular replacement using the structure of Myxococcus xanthus nucleoside diphosphate (NDP) kinase. NM23-H2 is a human NDP kinase. The enzyme catalyses phosphoryl transfer, binds DNA, and can activate the transcription of the c-myc oncogene in vitro. NM23 has also been reported to be a suppressor of metastasis in some types of tumours. Whereas the M. xanthus NDP kinase is a tetramer, NM23-H2 is a hexamer. The fold of NM23-H2 is identical to the fold of other NDP kinases. Two antiparallel helices joined by a turn form one edge of the nucleotide binding cleft. This region moves in a hinge-like fashion in response to substrate binding and crystal packing forces. Additional differences in conformation among the NDP kinases are principally in regions involved in protein-protein contacts within the oligomers. The only protein-protein interaction conserved among all NDP kinases is a dimeric interaction. Several mutations of NM23-H2 have been detected in tumour tissues. These mutations do not involve residues interacting with the substrates, and probably destabilise the enzyme without directly affecting the catalytic activity. Low level phosphorylation of serines has been reported for NM23 both in vitro and in vivo. The structure of the hexamer indicates that two serine residues that have been reported as being phosphorylated, Ser44 and Ser122, are on the surface of the hexamer, and are likely to be phosphorylated by exogenous kinases. In contrast, Ser120 is buried, and is most likely phosphorylated by a direct transfer from the phosphohistidine intermediate of the reaction mechanism.

The 1.9 A crystal structure of a nucleoside diphosphate kinase complex with adenosine 3',5'-cyclic monophosphate: evidence for competitive inhibition.

The X-ray structure of Myxococcus xanthus nucleoside diphosphate (NDP) kinase complexed with adenosine 3',5'-cyclic monophosphate (cAMP) has been determined. The structure was solved by difference Fourier analysis. The refined structure has a crystallographic R-factor of 0.17 at 1.9 A resolution. The phosphoryl group and ribose moiety make extensive polar interactions with the protein, whereas the base interacts only with two hydrophobic residues. The comparison with the structure of the enzyme complex with the substrate adenosine diphosphate (ADP) reported earlier shows that cAMP and ADP interact similarly with the enzyme. The base of the cAMP is present in two conformations, syn and anti, with respect to the sugar. The syn conformer is dominant. Based on the effect of cAMP on phosphorylation of the human NDP kinase NM23, it had been proposed that cAMP might interact with NDP kinase in a manner distinct from other nucleotides. However, the structure of the M. xanthus NDP kinase/cAMP complex indicates that the nucleotide is a competitive inhibitor of the enzyme and occupies the usual nucleotide site. Kinetic assays of the NDP kinase activity in the presence of cAMP were done. Their results are consistent with a competitive character of the cAMP inhibition.

Crystal structure of a diabody, a bivalent antibody fragment.

BACKGROUND: Diabodies are dimeric antibody fragments. In each polypeptide, a heavy-chain variable domain (VH) is linked to a light-chain variable domain (VL) but unlike single-chain Fv fragments, each antigen-binding site is formed by pairing of one VH and one VL domain from the two different polypeptides. Diabodies thus have two antigen-binding sites, and can be bispecific. Direct structural evidence is lacking for the connections and dimeric interactions between the two polypeptides of the diabody. RESULTS: The 2.6 A resolution structure has been determined for a bivalent diabody with a flexible five-residue polypeptide linker between the (amino-terminal) VH and (carboxy-terminal) VL domains. The asymmetric unit of the crystal consists of four polypeptides comprising two diabodies; for one of these polypeptides the linker can be traced between the VH and VL domains. Within each diabody the two associated VH and VL domains make back-to-back interactions through the VH domains, and there is an extensive VL-VL interface between the two diabodies in the asymmetric unit. CONCLUSIONS: The structure of the diabody is very similar to that which had been predicted by molecular modelling. Diabodies directed against cell-surface antigens should be capable of bringing together two cells, such as in cell-targeted therapy, because the two antigen-binding sites of the diabody are at opposite ends of the molecule and separated by approximately 65 A.

Evaluation of an enzyme-linked immunosorbent assay for detection of antibodies to vesicular stomatitis virus in cattle in an enzootic region of Mexico.

An ELISA was compared with the plaque-reduction serum neutralization (PRSN) test, for detection of vesicular stomatitis virus (VSV) antibodies in cattle in a vesicular stomatitis enzootic region of Mexico. A total of 325 bovine serum samples were screened for VSV antibodies. The PRSN test was performed, using Vero cells. The ELISA contained gradient-purified VSV Indiana (Lab strain) and VSV New Jersey (Hazelhurst) as the antigens. Regression analysis and weighted kappa statistic were used to estimate measures of agreement between the 2 assays for detection of VSV antibodies. The ELISA method proved useful for serodiagnosis of vesicular stomatitis. The ELISA and PRSN test results were highly correlated for detection of VSV antibodies.

Educational initiatives for cancer nursing in Europe.

The European Communities' "Europe against Cancer" programme proposed that Member States should recognize the specialist nature of oncology and recommended a plan of action relating to the training of health workers in cancer (Action 51 Europe Against Cancer Programme 1987-1989). The Advisory Committee on Training in Nursing arranged a review of training for nurses in cancer to be undertaken throughout Europe. One of the proposals formulated from the findings of this report was to produce a core curriculum and subsequent additional curricula in cancer for trained nurses throughout Europe. Following a workshop of European nurse educators and practitioners early in 1989, a core curriculum was produced by the European Oncology Nursing Society (EONS) and was published in September 1989. This paper outlines the content of that core curriculum and presents EONS' subsequent plans to design educational courses in all aspects of cancer care for European nurses. These courses will be supported by written and audiovisual materials, which may also be used for home learning.

Potential for insect transmission of HIV: experimental exposure of Cimex hemipterus and Toxorhynchites amboinensis to human immunodeficiency virus.

Human immunodeficiency virus (HIV) was detected in bedbugs (Cimex hemipterus) up to 8 d after oral exposure to highly concentrated virus in blood meals, but no virus replication was observed. HIV did not replicate in either intraabdominally inoculated bedbugs or intrathoracically inoculated mosquitoes (Toxorhynchites amboinensis). The virus was not detected in bedbug feces. Mechanical transmission of HIV by bedbugs could not be demonstrated in an in vitro model. The persistence of HIV in an insect or on its mouthparts is one of many factors necessary for mechanical transmission in nature. The risk of insect transmission of HIV appears to be extremely low or nonexistent.

Serum neutralizing antibody titers in dairy cattle administered an inactivated vesicular stomatitis virus vaccine.

Two doses of a formalin-killed, cell culture-derived vesicular stomatitis virus (vsv)-New Jersey serotype vaccine were administered intramuscularly, 30 days apart, to all lactating and nonlactating cows in a 350-cow dairy herd. Serum specimens were obtained serially from 96 cows before vaccination and at 30, 52 and 80 days after vaccination and from 24 of these cows 175 days after vaccination. Serum neutralizing antibody titers to vsv-New Jersey serotype were determined from serum-dilution, plaque-reduction tests. Serum neutralizing antibody titers also were determined during the same period for 67 nonvaccinated heifers in the herd. Peak group geometric mean serum neutralizing antibody titers of 1:530.46 +/- 1.14 (group geometric mean titer log10, 2.725 +/- 0.055) developed 21 days after the second vaccination, but decreased to a low value of 1:65.36 +/- 1.38 (group geometric mean titer log10, 1.815 +/- 0.142) by 175 days after vaccination. The nonvaccinated group had no detectable antibody titer to vsv-New Jersey serotype throughout the study. All serum specimens from the vaccinates and controls were negative for heterologous reactivity to vsv-Indiana serotype.

Epizootic vesicular stomatitis in Colorado, 1982: some observations on the possible role of wildlife populations in an enzootic maintenance cycle.

Sera obtained from wild ungulates, carnivores, and rodents in Colorado were tested for neutralizing (N) antibody against vesicular stomatitis, New Jersey serotype (VSNJ), virus to determine their involvement in the 1982 Colorado VSNJ epizootic in domestic animals. Viremic and N antibody responses of two local rodent species to a 1982 Colorado isolate of VSNJ were determined in the laboratory. The rodents produced only weak viremias, but all developed N antibody. N antibody prevalences for VSNJ in sera from wild ungulates was sufficiently high to indicate their involvement during the epizootic. In addition, the demonstration of N antibody in elk (Cervus elaphus) and mule deer (Odocoileus hemionus) prior to the epizootic in cattle and horses suggests that an enzootic cycle may exist in Colorado.

A prospective study of the epidemiology and ecology of Lassa fever.

In studies of Lassa fever in Sierra Leone, the prevalence of human antibody to Lassa virus ranged from 8% to 52%. Mastomys natalensis, the reservoir of Lassa virus, constituted 50%-60% of the rodents captured in houses but only 10%-20% of those captured in surrounding agriculture and bush areas (chi 2 = 90.2, P less than 10(-6), df = 1), a finding suggesting that houses are the most-important location for transmission of Lassa virus. Viral infection of Mastomys from houses ranged from 0% to 80%. The incidence of seroconversions in susceptible persons ranged from 5% to 22% per year; the ratio of illness to infection ranged from 9% to 26%, and the proportion of febrile illness associated with seroconversion was 5%-14%. Eightfold rises in titer of antibody occurred in 1%-18% of the antibody-positive population, a result suggesting reinfection. We estimate the ratio of fatalities to infection to be 1%-2%, a rate lower than estimates based on hospitalized cases. The high incidence of Lassa fever makes it a major problem in West Africa.

A case-control study of the clinical diagnosis and course of Lassa fever.

A prospective case-control study of Lassa fever was established in Sierra Leone to measure the frequency and case-fatality ratio of Lassa fever among febrile hospital admissions and to better delineate the clinical diagnosis and course of this disease. Lassa fever was responsible for 10%-16% of all adult medical admissions and for approximately 30% of adult deaths in the two hospitals studied. The case-fatality ratio for 441 hospitalized patients was 16.5%. We found the best predictor of Lassa fever to be the combination of fever, pharyngitis, retrosternal pain, and proteinuria (predictive value together, .81); of outcome, the best predictor was the combination of fever, sore throat, and vomiting (relative risk of death, 5.5). Complications included mucosal bleeding (17%), bilateral or unilateral eighth-nerve deafness (4%), and pleural (3%) or pericardial (2%) effusion. Lassa fever is endemic in this area and is a more-common cause of hospital admission and death than has previously been described; this disease must be considered when diagnosing febrile illness in West Africa.

Clinical virology of Lassa fever in hospitalized patients.

We measured levels of virus in sequential specimens from 137 patients with Lassa fever. The probability of fatal disease increased significantly with the level of viremia measured either on admission or during the course of illness. The odds ratio of death in patients with viremia greater than 10 TCID50/ml was 3.7 (90% confidence interval, 1.9-7.2). The same ratio in patients with viremia greater than 10 TCID50/ml and with levels of aspartate aminotransferase greater than or equal to 150 IU/liter was 21.5 (95% confidence interval, 5.2-99.0). Virus was found in throat cultures from 39% of viremic patients, compared with 14% of nonviremic patients (P less than .002); however, the level of virus was usually less than or equal to TCID50/ml. Fewer than 3% of patients were viruric during acute illness, and virus was isolated from three of three samples of cerebrospinal fluid. On admission, 53% of patients had IgG antibodies, and 67% had IgM antibodies. Recovery was not associated with the presence of either IgG or IgM. Virus was isolated from greater than 100 serum specimens that also contained high titers of IgG. Clinical Lassa fever was shown to be a disseminated systemic, primary viral infection, with an outcome highly associated with viremia but not with development of antibody.

Epizootic vesicular stomatitis in Colorado, 1982: infection in occupational risk groups.

In 1982-1983, an epizootic of vesicular stomatitis occurred in the western United States. Veterinarians, research workers, and regulatory personnel who were exposed to vesicular stomatitis virus were examined for patterns of human infection and prevalence of vesicular stomatitis New Jersey serotype neutralizing antibody. Insight into the mechanism of transmission was sought by comparing activities of antibody-positive and antibody-negative persons. A statistically significant risk factor was a history of infected animals sneezing in the face of serosurvey participants. Elevated odds ratios were also calculated for those who usually examined the oral cavity of affected animals, had open wounds on hands or arms, and had exposure to saliva through the eye or skin. Relatively intimate direct contact was required; a higher risk was associated with examining horses than cattle. Neutralizing antibody prevalence was significantly higher among exposed persons with illness (23%) than in exposed persons without a history of clinical illness (7%). Overall, however, infectivity of VSNJ for humans during the epizootic was low.

Epizootic vesicular stomatitis in Colorado, 1982: epidemiologic and entomologic studies.

An epizootic of vesicular stomatitis (VS) caused by the New Jersey serotype of VS virus affected livestock and humans in 14 western states in 1982-1983. Epidemiological observations were made on at least 10% of the cattle in 4 dairy herds that were located in the vicinity of Grand Junction, Colorado. High rates of neutralizing antibody to the New Jersey serotype were seen in all cattle regardless of whether livestock in the dairy had clinical VS or a decrease in mild production. Antibody titers remained high in these cattle for as long as 2 years after the epizootic. No virus isolations were made from 32 humans with clinical signs compatible with viral disease. Entomological information was obtained during the epizootic from 23 premises in northwestern Colorado. Insect collections yielded 4 isolates from Culicoides spp. midges, 2 from C. variipennis, and 1 each from C. stellifer and C. (Selfia) spp. This is the first report of VS virus isolations from field-collected Culicoides.

Epizootic vesicular stomatitis in Colorado, 1982: epidemiologic studies along the northern Colorado front range.

Epidemiologic evaluations were made of farm personnel on vesicular stomatitis-affected premises along the front range of the Rocky Mountains in Colorado during the 1982 epizootic. A similar antibody prevalence was noted to that of veterinarians and research and regulatory personnel who were involved with the same epizootic. Risk of infection resulted from intimate physical contact with infected horses or cows. Incidence and infection rates in horses were 45%; rates in cows were much lower, only 5%. Some epidemiologic clues were gained by a detailed study of an equine ranch. The pasture was incriminated as the area of highest risk, where 100% infection rates were noted. Horses in open pens and barns were at lower risk. Severe clinical disease in horses resulted in higher neutralizing antibody titers than inapparent or mild infection. Maternal antibody was detected in foals up to 4 months of age, and the level of antibody in the foal was a reflection of the dam's antibody level.

No evidence for increased risk of Lassa fever infection in hospital staff.

A prospective serological study was undertaken in hospital personnel who care for Lassa fever (LF) patients in an endemic region of Sierra Leone, West Africa. Among personnel from three hospitals where barrier nursing is practised, antibody prevalence and seroconversion by age and sex were consistently equal to or lower than those of persons in nearby village populations. No group among hospital personnel evaluated by age, sex, contact, or occupational exposure was at higher risk than another. Hospital staff in Sierra Leone who care for LF patients using simple barrier nursing methods have no higher risk of infection than the local population. These findings support the proposal that patients with LF in non-endemic countries need not be confined to isolators.