Preliminary observations on the pathogenesis of experimental bovine spongiform encephalopathy (BSE): an update

Further preliminary observations are reported of an experiment to examine the spread of infectivity and the occurrence of pathological changes in cattle exposed orally to infection with bovine spongiform encephalopathy. Calves were dosed at four months of age and clinically monitored groups were killed sequentially from two to 40 months after inoculation. Tissues were collected for bioassay, for histopathological examinations and for the detection of PrP. Previous reported observations have included the presence of infectivity in the distal ileum of cattle killed after six to 18 months, the earliest onset of clinical signs in an exposed animal after 35 months, and diagnostic histopathological changes in the brain, in association with clinical disease, after 36, 38 and 40 months. In spite of the relative inefficiency of the bioassay of scrapie-like agents across a species barrier the new observations confirm that the onset of clinical signs and pathological changes in the central nervous system (CNS) occur at approximately the same time. The earliest pathological change, the presence of abnormal PrP 32 months after inoculation, coincided with the earliest detected infectivity in the CNS and occurred shortly before there was evidence of typical spongiform changes in the brain 36 months after inoculation. Infectivity has now been demonstrated in the peripheral nervous system, in the cervical and thoracic dorsal root ganglia 32 to 40 months after inoculation and in the trigeminal ganglion 36 and 38 months after inoculation. At the time of writing evidence of infectivity in other tissues is confined to the distal ileum, not only after six to 18 months but also after 38 and 40 months, but these findings may be supplemented by the results of further mouse assays. Nevertheless, they are in general agreement with current knowledge of the pathogenesis of scrapie.     

Molecular analysis of bovine spongiform encephalopathy and scrapie strain variation

Five mouse scrapie strains, a mouse-passaged scrapie isolate derived from a field case in sheep in Germany, and 2 mouse-passaged bovine spongiform encephalopathy (BSE) isolates were analyzed by immunoblot in regards to banding patterns of proteinase K-digested pathologic prion proteins (PrPres). To obtain reliable results, the photo-imager technique was used for measurement of staining band intensities. Distinct and reproducible profiles were observed for the different strains or isolates. A British and a German BSE isolate were similar, suggesting the same source of infection. The German scrapie isolate resembled scrapie strain ME7, which has frequently been isolated from sheep scrapie in the past. In selected strains or isolates, no influence of the mouse lines used was observed on PrPres profiles, nor were brain region-specific differences apparent. This investigation suggests that PrPres glycotyping can be an invaluable tool for the in vitro differentiation of BSE and scrapie isolates.     

Neuropathological findings in cattle with clinically suspect but histologically unconfirmed bovine spongiform encephalopathy (BSE)

Neuropathological observations were made in 200 clinically suspected cases of bovine spongiform encephalopathy (BSE) in which pathognomonic vacuolar changes were absent. Routine histological and immunocytochemical techniques were applied to formalin-fixed, paraffin-embedded sections of the central nervous system. Significant neuropathological findings were detected in 85 (42.5 per cent) cases. The most frequent lesion, detected in 46 (23 per cent) cases, was a focal white matter vacuolation principally affecting the substantia nigra, but its clinical significance was unclear. Listeriosis was diagnosed in 17 (8.5 per cent) cases. In three of seven cases of non-suppurative encephalitis, lesions suggested sporadic bovine encephalomyelitis, a disease not previously reported in the UK. Suppurative thromboembolic or granulomatous lesions accounted for other inflammatory changes. Neuroectodermal tumours were present in five cases (2.5 per cent); three were identical in form and considered to be atypical ependymoma. Cerebrocortical necrosis, oedema or both were detected in four cases. The remaining cases (4.5 per cent), comprised those in which the changes were minor and of doubtful significance. Incidental pathological findings included occasional degenerating or vacuolated neurones, which occurred in the red nucleus in 105 brains, in the habenular nucleus in 71 brains, and singly at other sites in 17 brains. In sections of 37 brains immunostained with antiserum to prion protein (PrP), no evidence of PrP accumulation was found, providing some evidence that the series did not contain bovine prion disease cases which, based on the histological diagnosis, had given a false negative result. It is suggested that, of 115 cases (57.5 per cent) which lacked significant histological lesions, some were suffering from metabolic disorders. The study identified diseases and lesions which feature in the differential diagnosis of BSE. Their more accurate diagnosis may become particularly important if, as predicted, the BSE epidemic declines.     

182 offspring of cows with bovine spongiform encephalopathy (BSE) in Switzerland. 1. Clinical findings

This study involved 182 calves, heifers and cows that were the first generation progeny of cows with bovine spongiform encephalopathy (BSE). All animals underwent clinical and neurological examinations. Blood, milk, urine, ruminal fluid and cerebrospinal fluid samples were examined. Each cow underwent a specific examination for BSE which included assessment of behaviour, sensitivity and locomotion. This examination was performed twice, and there was very good agreement between the results. The most common symptom was nervousness (14 animals). In the first examination, 68 animals were mildly to moderately sensitive to touching of the head, 44 animals were hypersensitive to a halogen light and 35 animals were hypersensitive to a camera flash. Twenty-four animals reacted by kicking when the hindlimbs were touched with a broom. None of the animals had disturbances in locomotion. Based on the interpretation of all findings and the comparison of the results of the first and second examinations, 173 animals were diagnosed as free of BSE. In 9 animals, the disease could not be ruled out; however, it could not be confirmed in any of them. Histological and immunohistochemical examinations of the brain of all animals were negative for BSE (see Part 2 of the study for details). The results of this study indicate that abnormal clinical findings may occur in clinically healthy cattle. However, none of the progeny of cows with BSE exhibited clinical signs typical of BSE, such as disturbances in behaviour; sensitivity and locomotion.     

Effect of rendering procedures on the scrapie agent

A pool of scrapie-infected sheep brains was used to spike mixtures of porcine bone and intestine. These were processed in pilot-scale facsimiles of 12 rendering procedures that were in use within the European Union in 1991, and three that were not. Meat and bone meal, and tallow, were produced from the rendered tissues. Suspensions of all the meat and bone meal samples, and two of the tallow samples were assayed in mice for scrapie infectivity. Neither of the tallow samples had any detectable infectivity but the meat and bone meal samples were positive, except for those produced by processes involving exposure to hyperbaric steam. In addition, greaves were produced from the scrapie-spiked raw materials by an atypical low-temperature process and subjected to solvent extraction with hot heptane. The treated greaves were then exposed to steam to drive off residual solvent. Although the starting titre of infectivity in these greaves was low, there appeared to be no reduction in infectivity as a result of the treatments with hot heptane and steam. However, there was no detectable infectivity in the meat and bone meal prepared from the greaves produced by the atypical low-temperature process after it had been exposed to hyperbaric steam.     

Bovine spongiform encephalopathy (BSE): causes and consequences of a common source epidemic

Bovine spongiform encephalopathy (BSE) is a transmissible spongiform encephalopathy (TSE) or prion disease of cattle first recognized in 1986 in the United Kingdom, where it produced a common source epidemic that peaked in January 1993 and has subsided markedly since that time. The epidemic began simultaneously at many geographic locations and was traced to contamination of meat and bone meal (MBM), a dietary supplement prepared from rendering of slaughterhouse offal. It appears that the epidemic was initiated by the presence of the agent of scrapie (a long-standing TSE of sheep) that was first transmitted to cattle, beginning in the early 1980s, when most rendering plants abandoned the use of organic solvents in the preparation of MBM. The epidemic was probably accelerated by the recycling of infected bovine tissues prior to the recognition of BSE. To terminate the epidemic, a prohibition on the feeding of ruminant-derived protein to ruminants was introduced in the United Kingdom in July 1988. The ruminant feed ban accounts for the decline of the epidemic after an interval of about 5 years, approximately equivalent to the average incubation period of BSE. Relatively few cases of BSE have occurred in cattle born after 1993, and it is predicted that the epidemic will terminate about the year 2000 based on an extrapolation of the present declining curve. A comparison of data from the United Kingdom with data from relatively low incidence countries, such as Switzerland, indicates that this epidemic has been mainly confined to the United Kingdom because of a unique concatenation of risk factors, including: 1) a high ratio of sheep to cattle; 2) a relatively high rate of endemic scrapie; 3) the heavy feeding of MBM to dairy cattle; and 4) changes in the rendering process used to prepare MBM. Recently, cases of a variant form of Creutzfeldt-Jakob disease (a TSE of humans) have been reported in the United Kingdom. These cases, at least 10 of which had onset in 1994-1995, are distinguished by their occurrence in subjects under age 40 years, by their clinical presentation, and by their neurohistopathologic picture. The appearance of this novel disease and its concentration in the United Kingdom have raised the question that it might represent the transmission of BSE to humans. However, the cases gave no history indicating an unusual exposure to live cattle, to the preparation of beef products, or of dietary exposure to bovine tissues, and it remains to be determined whether they are associated with BSE.     

Bovine spongiform encephalopathy (BSE): clinical aspects, duty of notification and consequences of positive findings

Excimer lasers and high technology instrumentation have ushered in a new era of vision improvement surgery in Hawaii, replacing the more traditional forms of refractive surgery: cataract surgery, corneal transplant surgery, and radial keratotomy. Corneal surgery has been enhanced by new techniques of microsurgery and a more effective tissue procurement system for donor corneal tissue. Several laser centers provide the latest in FDA-approved excimer laser procedures including PRK and PTK. Mild to moderate myopia and astigmatism may now be corrected. Off-label use of LASIK, too, may soon be realized.     

The incidence of bovine spongiform encephalopathy in the progeny of affected sires and dams

Case control study techniques were used to compare the incidence of bovine spongiform encephalopathy (BSE) in the progeny of two affected sires and 110 affected dams with the incidence of BSE in the progeny of animals known to be unaffected at the last record. All the progeny were born before the ban on ruminant-derived protein in feedstuffs issued in July 1988. The results provide little, if any, evidence of differences between the incidence in the progeny of the affected animals and the incidence in the progeny of the presumed unaffected animals. Data from five herds were used in a logistic regression analysis to study the effects of the disease status of the dam and the age of the dam at the birth of the calf on the incidence of BSE. The disease status of the dam did not significantly affect the disease status of its progeny, after allowance had been made for the effects of herd, year and the age at last record of the progeny. The difficulty of establishing maternal transmission if a high proportion of the dams are incubating the disease and transmission can occur early in the incubation period is discussed.     

Alteration of rendering and meat hygiene legislation in the course of the BSE/TSE crisis. Validity of decisive legal provisions (1)

The decisive European legal provisions to fight bovine sponfiform encephalopathy in the context of the rendering and meat-hygiene legislation are the Commission Decisions 96/449/EC "on the approval of alternative heat treatment systems for processing animal waste with a view to the inactivation of spongiform encephalopathy agents" and 97/534/EC "on the prohibition of the use of material presenting risks as regards transmissible spongiform encephalopathies". Both decisions are based on Council Directives which do not provide the necessary authorization for these decisions by the Commission. Consequently, both have to be regarded as legally invalid. Furthermore, the legal acts to convert these decisions into German law (Rendering Plants Regulation and Meat-Hygiene Regulation) have to be considered invalid--partly due to formal and partly due to factual reasons. Because of the thus created legal uncertainty the legal provisions in question can only come fully into force after the elimination of this specifically described legal shortcomings or after clarification of the legal situation by the responsible courts.     

Prion protein immunohistochemical staining in the brains of monkeys with transmissible spongiform encephalopathy

BACKGROUND: Spirituality is receiving greater attention in the medical literature, especially in the family practice journals. A widely applicable instrument to assess spirituality has been lacking, however, and this has hampered research on the relationship between spirituality and health in the clinical setting. METHODS: A new instrument, called the Spiritual Involvement and Beliefs Scale, was designed to be widely applicable across religious traditions, to assess actions as well as beliefs to address key components not assessed in other available measures, and to be easily administered and scored. The instrument is a questionnaire containing 26 items in a modified Likert-type format. Following careful pretesting, the instrument was administered to 50 family practice patients and 33 family practice educators. The validity and reliability of the instrument were then evaluated. RESULTS: By several measures, instrument reliability and validity are very good, with high internal consistency (Cronbach's alpha = .92); strong test-retest reliability (r = .92); a clear four-factor structure; and a high correlation (r = .80) with another established measure of spirituality, the Spiritual Well-Being Scale. CONCLUSIONS: The Spiritual Involvement and Beliefs Scale (SIBS) appears to have good reliability and validity. Compared with other instruments that assess spirituality, the SIBS has several theoretical advantages, including broader scope, use of terms that avoid cultural-religious bias, and assessment of both beliefs and actions. More testing is underway to further assess its usefulness.     

Studies on the efficacy and safety of biapenem (L-627) against infections in pediatrics

The efficacy and the safety of biapenem (L-627), a new carbapenem antibiotic, against infections in pediatrics were studied. The obtained results are summarized as follows. 1. L-627 at dose levels of 5.4 mg/kg to 12.4 mg/kg (daily doses of 16.2 mg/kg to 37.2 mg/kg) was administered by intravenous drip infusion 3 times daily for 5 to 7 days to 2 cases of pneumonia, 3 cases of skin and soft-tissue infections and 1 case of urinary tract infection for a total of 6 cases. As results, all the cases showed good or better responses including 4 excellent and 2 good results. Bacteriological efficacies in all of the 3 eligible cases were assessed as "eradicated". 2. As for the safety, a decrease in the WBC count and slight elevation of GOT and GPT were observed in 1 case as abnormal changes in the laboratory tests results, only incidence of side effect observed was the eruption in 1 case. 3. The results above indicate that L-627 is useful for the treatment of general infections in pediatrics.     

Studies on the desamidation of bovine collagen

A variable air-volume, parallel-plate extrapolation chamber forming an integral part of a Solid Water phantom was built to determine the absorbed dose in Solid Water directly. The sensitive air-volume of the extrapolation chamber is controlled through the movement of the chamber piston by means of a micrometer mounted to the phantom body. The relative displacement of the piston is monitored by a calibrated mechanical distance travel indicator with a precision on the order of 0.002 mm. Irradiations were carried out with cobalt-60 gamma rays, x-ray beams ranging from 4 to 18 MV, and electron beams between 6 and 22 MeV. The absorbed dose at a given depth in Solid Water is proportional to the ionization gradient measured in the Bragg-Gray cavity region with an extrapolation chamber embedded in the Solid Water phantom. The discrepancies between the doses determined in Solid Water with our uncalibrated extrapolation chamber and doses obtained with a calibrated standard thimble ionization chamber are at most 1% for photon and electron beams at all megavoltage clinical energies. Uncalibrated extrapolation chamber thus offer a simple and practical alternative to other techniques used in output measurements of megavoltage photon and electron machines.     

Therapeutical approaches to transmissible spongiform encephalopathies (TSE): the case of amphotericin-B

Within 1987-1995 the authors observed 16 cases of tuberculosis in HIV-infected patients which accounted for 26.7% of AIDS patients treated by them. 14 cases were diagnosed intravitally, 2 postmortem. Infiltrative, generalized, cavernous, intrathoracic lymph node, intraabdominal lymph node tuberculosis and tuberculous pleurisy were identified in 5, 6, 2, 1, 1 and 1 patients, respectively. 6 patients from the above are still alive and are receiving treatment (5 of them with infiltrative tuberculosis), 10 died. Tuberculosis course and outcomes in HIV-infected subjects depended on the stage of their immunodeficiency. In moderate immunodeficiency (CD4-lymphocyte > 200/mm3) tuberculosis ran, as a rule, as local and infiltrative, sensitive to specific therapy. In severe damage to immune system (CD4 < 100/mm3) tuberculosis acquired a generalized course, sometimes fulminant, resistant to treatment. It is inferred that HIV-infected subjects with immunodeficiency need tuberculosis prophylaxis with isoniazide or rifampicin.     

Studies on the efficacy of child psychoanalysis.

This article summarizes 3 studies that evaluated the psychoanalytic psychotherapeutic treatment of diabetic children and adolescents with grossly abnormal blood glucose profiles necessitating repeated admissions to hospital. Study 1 used time series analysis to demonstrate that improvements in control were predicted by unconscious themes emerging in the analytic material. Study 2 compared the effect of psychotherapeutic treatment with that of minimal psychological intervention in 2 well-matched groups (n?=?11). Patients in the treatment group were offered psychoanalytic psychotherapy 3–4 times per week on the hospital ward. The intervention was highly effective in improving the diabetic control of the children, and this improvement was maintained at 1-yr follow-up. Study 3 used single-case experimental design to demonstrate the marked effect of psychotherapeutic help on growth in diabetic children with short stature. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

New agents for in vivo chelation of uranium(VI): efficacy and toxicity in mice of multidentate catecholate and hydroxypyridinonate ligands

Soluble uranyl ion [UO2(2+), U(VI)] is a kidney poison. Uranyl ion accumulates in bone, and the high specific activity uranium isotopes induce bone cancer. Although sought since the 1940's, no multidentate ligand was identified, until now, that efficiently and stably binds U(VI) at physiological pH, promotes its excretion, and reduces deposits in kidneys and bone. Ten multidentate ligands patterned after natural siderophores and composed of sulfocatechol [CAM(S)], carboxy-catechol [CAM(C)], or hydroxypyridinone [Me-3,2-HOPO] metal-binding units have been tested for in vivo chelation of U(VI). Ligands were injected intraperitoneally (i.p.) into mice 3 min after intravenous (i.v.) injection of 233U or (232+235)U as UO2Cl2 [ligand-to-metal molar ratio 75 to 92]. Regardless of backbone structure, denticity, or binding unit, all 10 ligands significantly reduced kidney U(VI) compared with controls or with mice given CaNa3-DTPA, and four CAM(S) or CAM(C) ligands also significantly reduced skeleton U(VI). Several ligands removed U(VI) from kidneys, when injected at 1 or 24 h. Injected at molar ratios > or = 300, 5-LIO(Me-3,2-HOPO) and TREN-(Me-3,2-HOPO) reduced kidney U(VI) to about 10% of control. Given orally to fasted mice at molar ratios > or = 300, those ligands significantly reduced kidney U(VI). In mice injected i.v. with 0.42 micromol kg(-1) of 235U and given 100 micromol kg(-1) of one of those Me-3,2-HOPO ligands i.p. daily for 10 d starting at 1 h after the U(VI)) loss of kidney U(VI) was greatly accelerated, and the kidneys of treated mice showed no microscopic evidence of renal injury. Crystals of uranyl chelates with linear tetradentate ligands containing bidentate Me-3,2-HOPO groups demonstrate a 1:1 structure. Considering low toxicity, effectiveness, and reasonable cost, the structurally simple linear tetradentate ligands based on the 5-LI backbone (diaminopentane) offer the most promising approach to a clinically acceptable therapeutic agent for U(VI). Work is in progress to identify the most suitable CAM or HOPO binding unit(s).     

Clinical efficacy of lactulose in cirrhotic patients with and without subclinical hepatic encephalopathy

Seventy-five cirrhotic patients with hyperammonemia in the past or at the time of the study were randomly divided into two groups (treated with lactulose or nontreated) in 14 hospitals in Japan. Thirty-six cirrhotic patients were diagnosed as having subclinical hepatic encephalopathy (SHE), and 39 were diagnosed as non-SHE. SHE was diagnosed when the results of all three of the quantitative psychometric tests used (number connection test, and symbol digit and block design tests of the Wechsler adult intelligence scale [revised]) were abnormal as compared with age-matched normal values. The mean number of abnormal psychometric test results and the prevalence of SHE were used for a quantitative evaluation of the efficacy of the lactulose treatment. Twenty-two of the SHE patients were treated with lactulose (45 mL/d) for 8 weeks, and the other 14 SHE patients did not receive lactulose. In the SHE patients administered lactulose, the results of the quantitative psychometric evaluation were significantly improved at 4 and 8 weeks after the beginning of the lactulose administration. The SHE had disappeared in 10 (50%) of the 20 treated patients at week 8, but it persisted in 11 (85%) of the untreated 13 patients. We concluded that lactulose treatment in cirrhotic patients with SHE is effective with respect to psychometric tests.     

Brainstem auditory evoked potentials (BAEPs) and somatosensory evoked potentials (SEPs) in enteric encephalopathy (EE)

BAEPs and SEPs were studied in 25 patients of enteric encephalopathy in acute phase and the results were compared with 25 healthy control persons. In the study the important observations of BAEPs were delayed peak latency of wave III, wave V and delayed ILP I-V, and of SEPs was prolonged peak latency of N20. The electrophysiological evidence suggests metabolic cause for the coma and the SEP changes were similar to those observed in cerebral malaria reported earlier in this laboratory.