Delivery of melanoma-associated immunoglobulin monoclonal antibody and Fab fragments to normal brain utilizing osmotic blood-brain barrier disruption.

Iodinated monoclonal antibodies (IgG 96.5 and two monomeric Fab fragments 96.5 and 48.7) to melanoma-associated antigens were administered after osmotic blood-brain barrier (BBB) opening in normal rats. Osmotic BBB disruption significantly (P less than 0.0001) increased monoclonal antibody delivery to the brain. Following BBB opening and intracarotid administration, there was no difference in the disrupted brain concentration integral area under the curve between Fab and IgG over the 72-h experimental period. However, Fab concentration in the disrupted brain was initially higher than IgG, and the clearance was more rapid (P less than 0.0001), decreasing 50% by approximately 4.5 h compared to 25.5 h for IgG. Plasma clearance was also more rapid for the Fab than IgG. The levels decreased 50% by 1.5 h for Fab and 15 h for IgG. The route and timing of antibody infusion had a significant effect on delivery to the disrupted brain with the Fab fragments but not with the intact IgG. Antibody recovered from disrupted brain retained its immunological reactivity as measured by a cell binding assay for at least 24 h. IgG and Fab delivery to the ipsilateral brain after BBB disruption increased (P less than 0.001) with increasing dose over a more than 3-log dose range. These data provide information applicable to the therapeutic use of monoclonal antibodies in brain tumor treatment.

Histomorphometric effects of calcium or calcium plus 25-hydroxyvitamin D3 therapy in senile osteoporosis.

To evaluate the effects of calcium and 25-OHD in the therapy of senile osteoporosis, we studied a group of 39 women aged 69 +/- 7 (standard deviation, SD) years with severe osteoporosis. The group was characterized histomorphometrically by depressed bone remodeling rates without hyperosteoidosis. No subject had risk factors for osteopenia other than their age and postmenopausal status, and no subject was receiving therapy for bone disease at the onset of the study. Subjects were followed for 2 years after randomization to receive either 1200 mg/day of calcium (as calcium carbonate) and 40 micrograms/day of 25-OHD (calcium-25-OHD group), or 1200 mg/day of calcium plus placebo (calcium-placebo group). Calcium-25-OHD resulted in a clear increase in 25-OHD levels (p less than 0.001) and an increase in calcium absorption as indicated by urinary calcium excretion. Nevertheless, there was no significant change in fasting serum calcium, phosphorus, alkaline phosphatase, PTH, or 1,25-(OH)2D in either group. Radial and phalangeal bone mineral content and trabecular bone volume in the biopsied patients remained stable in both groups over the 2 year period. Unexpectedly, repeat bone biopsies revealed a clear improvement in the rate of mineralization in both groups, presumably as a result of the calcium supplementation alone. In summary, calcium-placebo and calcium-25-OHD treatment were both associated with stable appendicular bone mineral content in women with senile osteopenia. The finding of an effect of calcium supplementation on the rate of mineralization indicates that relative calcium deficiency may impair the mineralization phase of remodeling.(ABSTRACT TRUNCATED AT 250 WORDS)

Calcium and cholecalciferol: effects of small supplements in normal men.

The effect of small calcium and vitamin D supplements on mineral metabolism in normal persons is unclear. To investigate the biochemical response to these medications, we administered 1000 mg Ca and 25 micrograms cholecalciferol per day or a placebo to 92 normal men for 1 y. The Ca and cholecalciferol were tolerated well. 25-Hydroxycholecalciferol [25-(OH)D] and 24,25-dihydroxycholecalciferol [24,25-(OH)2D] levels rose in treated subjects; there was no definite change in 1,25-(OH)2D concentrations. The average difference in 25-(OH)D levels between treated and untreated subjects was 30 nmol/L at 1 y. Fasting serum Ca, alkaline phosphatase, creatinine, and parathyroid hormone levels and the fasting urinary excretion of Ca, phosphorus and cAMP, were not affected. However, 24-h urinary Ca excretion was higher in the supplemented group (3.5 +/- 1.9 vs 4.7 +/- 1.7 mmol/d, p = 0.006). Serum P concentrations were slightly higher in the supplemented group at 1 y. In normal men small calcium and cholecalciferol supplements are safe, provide adequate vitamin D nutrition and apparently increase net gastrointestinal Ca absorption.

Serum protein concentrations and bone mineral content in aging normal men.

Low serum protein levels and low dietary protein intakes possibly influence bone metabolism. To evaluate the relationships among serum protein concentrations, protein intakes, and bone mineral content (BMC), we have examined two populations of normal men. BMC was measured at two radial sites and a vertebral site. Aging was associated with a fall in BMC at all sites. Serum albumin levels also fell with age and were related to BMC. Albumin concentrations were associated with BMC when other variables (calcium and protein intake, vitamin D levels, parathyroid hormone, and urinary calcium) also were considered. Thus, alterations in protein metabolism may affect BMC and may play a role in the genesis of senile osteopenia.

Long-term intake of soy protein improves blood lipid profiles and increases mononuclear cell low-density-lipoprotein receptor messenger RNA in hypercholesterolemic, postmenopausal women.

The long-term clinical effects of soy protein containing various amounts of isoflavones on lipoproteins, mononuclear cell LDL receptor messenger RNA concentrations, and other selected cardiovascular risk factors are not well known. Sixty-six hypercholesterolemic, free-living, postmenopausal women were investigated during a 6-mo parallel-group, double-blind trial with 3 interventions. After a control period of 14 d, all subjects were randomly assigned to 1 of 3 dietary groups (all with 40 g protein): a National Cholesterol Education Program (NCEP) Step 1 diet with protein from casein and nonfat dry milk (control), an NCEP Step 1 diet with protein from isolated soy protein containing moderate amounts of isoflavones (ISP56), or an NCEP Step 1 diet with protein from isolated soy protein containing high amounts of isoflavones (ISP90). Non-HDL cholesterol in both the ISP56 and ISP90 groups was reduced compared with the control group (P < 0.05), whereas total cholesterol was not changed. HDL cholesterol increased in both the ISP56 and ISP90 groups (P < 0.05), whereas the ratio of total to HDL cholesterol decreased significantly in both groups compared with the control (P < 0.05). Mononuclear cell LDL receptor messenger RNA concentrations increased in subjects consuming ISP56 or ISP90 compared with the control (P < 0.05). These results indicate that soy protein, with different amounts of isoflavones, may decrease the risk of cardiovascular disease via improved blood lipid profiles, and that the mechanism by which apolipoprotein B-containing lipoproteins were depressed may be via alterations in LDL receptor quantity or activity.

Risk factors for drug-induced parkinsonism in tardive dyskinesia patients.

Using multivariate statistical analyses, the authors identified risk factors for development of drug-induced parkinsonism (DIP) in 66 tardive dyskinesia (TD) patients. Older age, recent use of neuroleptics, shorter duration of past neuroleptic exposure, and severity of TD were associated with increased risk of DIP. The clinician should devise treatment strategies in anticipation of the occurrence of DIP regardless of the presence or absence of TD, especially in older patients. New models for the pathophysiology of the two disorders are needed.

Application of clinically valid cardiac risk factors to aortic aneurysm surgery.

The operative outcome of 97 consecutive nonruptured infrarenal aortic aneurysms is analyzed regarding clinically identifiable cardiac risk factors. Clinically evident coronary artery disease was present in 45 patients (46%). Operative mortality was 4% (four cardiac deaths) with an additional 4% nonfatal postoperative myocardial infarction rate. All cardiac complications occurred in patients with clinically evident coronary artery disease, while no mortality occurred in 52 patients lacking a preoperative history of myocardial infarction, congestive heart failure, or angina. Preoperative risk factors having a significant negative influence on outcome include a history of prior myocardial infarction and compensated congestive heart failure. Few patients with aneurysms who have clinical evidence of coronary artery disease are indicated for coronary arteriography and bypass prior to aneurysm repair. Furthermore, indications for invasive cardiac screening of the patient with an aneurysm who lacks cardiac symptoms are limited.

Risk factors for infection with Toxoplasma gondii for residents and workers on swine farms in Illinois.

Risk factors for Toxoplasma gondii infection in workers and residents of swine farms were studied on 43 farms in Illinois. Blood samples were collected from 174 adults in 1993. The T. gondii seroprevalence was 31%. An interview was conducted with each participant, obtaining information on demographic characteristics and behaviors suspected to affect the risk of T. gondii infection. Factors associated with increased risk of T. gondii seropositivity were a higher number of seropositive cats trapped on the farm, male sex, rearing pigs on pasture, and gardening. Factors associated with a decreased risk were handling of pig feed and presence of cats inside the pig facilities. Thus, infection of cats with T. gondii increased the risk of human infection, and contact with soil was a likely mechanism for transmission. The increased risk of seropositivity in males is attributed to less attention paid to cleanliness in food preparation and eating.

Social networks and negative versus positive symptoms of schizophrenia.

The social networks of people with schizophrenia are less functional than those of nonpsychiatric populations. In this study, positive versus negative symptoms were examined in relation to social networks. Thirty-nine patients with chronic schizophrenia were assessed for positive and negative symptoms and for size and quality of social network. Patients with more negative symptoms had significantly smaller social networks. Their networks were similarly more dysfunctional on a variety of measures. Positive symptoms did not correlate significantly with any network variable.

Underrecognition of tardive dyskinesia and drug-induced parkinsonism by psychiatric residents.

Recognition of tardive dyskinesia (TD) and other neuroleptic, drug-induced, extrapyramidal side effects presents a major challenge in modern clinical psychopharmacology. Failure to recognize these disorders can lead to poor patient care and may contribute to societal pressure for external control of psychiatric practice. This study reports the occurrence of tardive dyskinesia and drug-induced parkinsonism (DIP) in 101 inpatients, and documents underrecognition of both disorders by resident physicians. Researchers noted TD in 28% of cases and residents only described TD (or symptoms of TD) in 12%. The researcher determined DIP prevalence rate of 26% contrasted with an 11% rate found by residents. Patients with psychotic disorders were more likely than other patients to have researcher-identified TD, whereas DIP (researcher cases) occurred more often in patients with affective diagnoses. Residents tended to miss milder cases of TD, and to miss DIP in younger patients and in patients with affective disorders. Improved teaching and clinical exams are recommended to improve recognition.

Contributions of exercise, body composition, and age to bone mineral density in premenopausal women.

The purpose of this cross-sectional study were to determine whether exercisers have greater bone mineral density (BMD) than nonexercisers, whether aerobic dancers have greater BMD than walkers, and to determine the contributions of energy expenditure, body composition, and dietary factors to spine and femur BMD. Measurements were obtained on 93 eumenorrheic women (walkers N = 28; aerobic dancers, N = 34; nonexercisers, N = 31) ages 25-41 yr; lumbar spine and proximal femur BMD, body composition, physical activity, and nutrient intakes. Mean height, weight, and body mass index and median age and calcium intakes were similar for the three groups. Mean (+/- SD) values of the spine, total femur, and femoral neck BMD, respectively, were: walkers (1.092 (+/- 0.098), 0.947 g.cm-2), dancers (1.070 (+/- 0.124), 0.990 (+/- 0.104), 0.908 (+/- 0.106) g.cm-2), and nonexercisers (1.020 (+/- 0.112), 0.887 (+/- 0.073), 0.792 (+/- 0.089) g.cm-2) multiple regression analyses indicated that exercise contributed to spine (P = 0.018), total femur (P =0.012), and femoral neck (P < 0.0001) BMD, whereas type of exercise (aerobic dance vs walking) did not (P > 0.05). Total femoral BMD was influenced by exercise (P = 0.012) and energy expenditure (P = 0.023), while vertebral BMD was influenced by age (P = 0.0067), body weight (P = 0.017), and exercise (P = 0.018). These findings suggest that walking and aerobic dance exercise may provide physically active premenopausal women with greater lumbar and femoral BMD than sedentary females.

Sensitivity of the standardized pseudorabies virus neutralization test varies with the test strain used.

The effect of altering the strain of the test virus used in the standardized pseudorabies virus neutralization (VN) test on the sensitivity of the assay was evaluated. Comparative VN tests were performed using 4 different strains: the avirulent Bartha parental, the avirulent recombinant Bartha gIIIKa, the moderately virulent Shope (currently used for the VN test at the National Veterinary Services Laboratory, Ames, IA), and the highly virulent P2208 (Funkhauser). A radioimmunoassay and a Western immunoblotting technique were employed to verify the presence of anti-pseudorabies virus (PrV) antibodies in sera. Statistical analysis indicated that replacement of the Shope strain by the Bartha gIIIKa or the P2208 strain resulted in VN titers that were 4.23- and 2.00-fold higher, respectively. Despite these differences, specificity with regard to PrV diagnosis was unaltered. This apparent enhancement of the sensitivity of the PrV VN test would be beneficial for the serologic identification of PrV-infected animals during an eradication effort.

Evaluation of the sensitivity and specificity of two diagnostic tests for antibodies to pseudorabies virus glycoprotein X.

The diagnostic performance of 2 enzyme-linked immunosorbent assays (gX-T, gX-H) for antibodies to pseudorabies virus (PRV) glycoprotein X (gX) were evaluated using 311 serum samples from a nonvaccinated quarantined herd. When the standardized virus neutralization (VN) test, which uses the Shope strain (VN Shope), was used as the comparative diagnostic standard, the gX-T test had a 7% false-negative rate and a 52% false-positive rate, and the gX-H test had a 19% false-negative rate and a 19% false-positive rate. When the VN test with a Bartha recombinant strain (VN Bartha gIIIKa) was used as the diagnostic standard, the gX-T test had a 9% false-negative rate and a 26% false-positive rate, and the gX-H test had a 24% false-negative rate and a 11% false-positive rate. Thus, the gX-T test was more sensitive and the gX-H test was more specific. Additional diagnostic tests on 79 serum samples from a noninfected herd did not produce false positives for the gX-H test, but there was an 8% false-positive rate for the gX-T test. Previous studies from our laboratory have demonstrated that VN Bartha gIIIKa has higher sensitivity than VN Shope, without losing specificity, and thus is a better comparative diagnostic standard. When adding a suspect range to the gX-T test, using the same criteria as the suspect range for the gX-H test, the false-positive rate of the gX-T test was reduced to 5% when evaluated versus VN Bartha gIIIKa in the infected herd and to 1% for the PRV-negative herd.(ABSTRACT TRUNCATED AT 250 WORDS)

Application of a quantitative algorithm to restriction endonuclease analysis of Aujeszky's disease (pseudorabies) virus from a geographically localized outbreak.

A retrospective epidemiologic study was conducted to evaluate the application of an objective quantitative algorithm for estimating genetic similarity from restriction endonuclease analysis data. The analysis was performed to assist the determination of chronologic trends in an Aujeszky's disease viral epidemic in a geographic region. DNA from each viral isolate obtained during the epidemic was digested with 4 restriction endonucleases and molar ratio labeled to generate separate fragment patterns that were simultaneously compared using the algorithm. The resultant estimates of genetic similarity were then used in conjunction with time of virus isolation and specific geographic location of the outbreaks to identify the probable sources of infection and the patterns of spread among swine herds. This type of quantitative analysis enabled a more precise and objective approach than previously has been applied to the interpretation of restriction endonuclease data, thereby demonstrating the benefit of this methodology for the investigation of infectious disease outbreaks.

Quantitative assessment of genomic similarity from restriction fragment patterns.

A quantitative algorithm for comparing restriction fragment patterns (RFPs) was developed and used to estimate the genomic similarity of 18 isolates of pseudorabies virus of known origin. Variants of this algorithm using either untransformed or square-root-transformed differences between fragment sizes were investigated with regard to their ability to classify RFPs. Multidimensional scaling was used to represent spatially the genomic relatedness among samples, with 3 dimensions producing the most meaningful results. The square-root transformation provided more interpretable dimensions. The first dimension differentiated samples geographically, separating North American from European isolates. The second and third dimensions differentiated isolates with specific gene deletions (gE and gG, respectively) from those not having these deletions. Clusters of isolates were identified that were related either by collection from the same geographic area during a specific time period, or by laboratory intervention to create vaccines. These methods offer increased precision in the determination of genetic relatedness based on RFPs, and thus offer increased diagnostic accuracy for the determination of sources of infection.

Herd factors affecting the selection and success of intervention strategies in the program for eradication of pseudorabies (Aujeszky's disease) virus from Illinois swine farms.

The program for eradication of pseudorabies virus (PrV) from swine herds in Illinois was evaluated with respect to compliance with Livestock Conservation Institute (LCI) guidelines for selection of intervention strategies and for the effectiveness of these interventions under different herd conditions. The sample consisted of 395 swine operations quarantined between 1988 and 1994. These herds were followed until the end of 1996. The association of herd characteristics (number of sows, sow PrV seroprevalence, type of housing, number of PrV-seropositive farms within 1.5 mi) at the time of quarantine with the producer's selection of an initial intervention strategy (vaccination, offspring segregation, test-and-removal, depopulation-and-repopulation) was analyzed using logistic multiple regression. The interaction of herd characteristics with intervention strategies to affect the duration of quarantine was analyzed using multivariable Cox regression. Factors favoring selection of vaccination were increased herd size, higher sow PrV seroprevalence, and more PrV-seropositive farms within 1.5 mi. Offspring segregation was preferred when sow PrV seroprevalence was higher, and test-and-removal was preferred when seroprevalence was lower. Depopulation-and-repopulation was more likely in outdoor operations. Except for depopulation-and-repopulation, selection of an intervention strategy was in accordance with LCI guidelines. Vaccination and offspring segregation were associated with longer times under quarantine, and test-and-removal and depopulation-and-repopulation with shorter times. Test-and-removal was more effective in reducing the duration of quarantine when sow PrV seroprevalence was low. Vaccination increased the duration of quarantine less when sow PrV seroprevalence was high. Vaccination increased the duration of quarantine more when there were one or more PrV-seropositive farms within 1.5 mi than when there were no PrV-seropositive farms within 1.5 mi. It is apparent that herd characteristics affect the duration of quarantine and therefore need to be taken into account in the selection of a PrV-intervention strategy.

Associations between genetics, farm characteristics and clinical disease in field outbreaks of porcine reproductive and respiratory syndrome virus.

Porcine reproductive and respiratory syndrome (PRRS) is a disease of domestic swine characterized by exceptionally high clinical variability. This study addresses the question of whether clinical variability in PRRS results from (a) genetic variation among viral isolates and/or (b) variation in management practices among farms on which isolates are found. Genetic data (open reading frame 5 gene sequences) and data on farm characteristics and associated clinical disease signs were collected for 62 PRRS virus (PRRSV) field isolates, representing 52 farms. Clinical disease signs were interrelated--confirming that a true reproductive syndrome exists (involving abortions, infertility in sows, deaths of sows and preweaning mortality). Pairs of farms experiencing deaths in their sow populations also tended to share viral isolates which were more similar to one another than expected by chance alone. This implies that sow death (one of the more-severe manifestations of PRRS) is under genetic influence. Large herd size was a significant risk factor for the death of sows and for respiratory disease in nursery pigs. All-in-all-out management practices in the nursery were protective against reproductive signs in the sow herd. All-in-all-out management practices in the finishing stages of production were protective against respiratory disease in nursery pigs--but were paradoxically associated with an increased risk of infertility in sows. These results suggest that farm-management practices can also influence which PRRS clinical signs are manifested during an outbreak. In general, signs associated with PRRS appear to result from a combination of genetic factors and herd-management characteristics. The relative contributions of these two influences differ depending on the specific clinical sign in question.

A field trial of the effectiveness of a feline Toxoplasma gondii vaccine in reducing T. gondii exposure for swine.

A 3-yr field trial was conducted on 8 commercial swine farms in Illinois to determine the effectiveness of a feline Toxoplasma gondii vaccine in reducing the exposure of swine to T. gondii. A vaccine consisting of live bradyzoites of the mutant T-263 strain, capable of preventing oocyst shedding by cats, was used in this study. Each farm was visited 3 times in 1994, 3 times in 1995, and once in 1996. Cats were trapped and inoculated with the T-263 oral vaccine during 1994 and 1995. On each visit, the following samples were collected: blood from pigs, cats, and mice for detection of serum antibodies to T. gondii, feces from cats to detect oocysts, and heart and brain tissues from rodents to determine the presence of T. gondii tissue cysts. The modified agglutination test (MAT), with a positive titer set at the 1:25 dilution, was used to determine serum antibodies. At first capture, 72.6% (61/84) of juvenile cats and 32.6% (31/95) of adult cats had no detectable antibodies (seronegative), indicating no prior exposure to T. gondii when they received their first vaccine. Of these first-time seronegative cats, 58.1% (18/31) of adult and 45.9% (28/61) of juvenile cats were recaptured and received a second dose of vaccine. Changes in the prevalence of T. gondii infection were evaluated from the prevaccination (1992, 1993) to the postvaccination (1996) period. Eleven cats (5%) were detected shedding oocysts between 1994 and 1996, of which 10 (90.1%) shed during 1994. The last detection of oocyst shedding by cats was during the first farm visit in 1995. There was a significant decrease in T. gondii seroprevalence for finishing pigs (P < 0.05, Wilcoxon sign rank test). There was a positive correlation (Spearman's p = 1.0, P < 0.0001) between the change in prevalence in juvenile cats and the change in prevalence in finishing pigs. The seropositivity rate (MAT > or = 1:25) in mice among all farms decreased from 4% in 1992-1993 to 0% in 1996. The mean prevalence of T. gondii tissue cyst isolation for mice on all farms decreased from 1.1% in 1994, to 0.8% in 1995, and to 0.5% in 1996. The results of this study suggest that the reduced exposure of pigs to T. gondii was due to the administration of the T. gondii vaccine to cats.

Sources and reservoirs of Toxoplasma gondii infection on 47 swine farms in Illinois.

Field studies were conducted on 47 swine farms in Illinois during 1992 and 1993 to identify sources and reservoirs of Toxoplasma gondii infection. Blood samples were obtained from swine and from trapped wildlife. Serum antibodies to T. gondii were determined using the modified agglutination test, incorporating mercaptoethanol. Antibodies to T. gondii (titer > or = 25) were found in 97 of 4,252 (2.3%) finishing pigs, 395 of 2,617 (15.1%) sows, 267 of 391 (68.3%) cats, 126 of 188 (67.0%) raccoons, 7 of 18 (38.9%) skunks, 29 of 128 opossums (22.7%), 6 of 95 (6.3%) rats, 3 of 61 (4.9%) white-footed mice (Peromyscus sp.), and 26 of 1,243 (2.1%) house mice (Mus musculus). Brains and hearts of rodents trapped on the farm were bioassayed in mice for the presence of T. gondii. Toxoplasma gondii was recovered from tissues of 7 of 1,502 (0.5%) house mice, 2 of 67 (3.0%) white-footed mice, and 1 of 107 (0.9%) rats. Feces of 274 cats trapped on the farm and samples of feed, water, and soil were bioassayed in mice for the presence of T. gondii oocysts. Toxoplasma gondii was isolated from 2 of 491 (0.4%) feed samples, 1 of 79 (1.3%) soil samples, and 5 of 274 (1.8%) samples of cat feces. All mammalian species examined were reservoirs of T. gondii infection. All farms had evidence of T. gondii infection either by detection of antibodies in swine or other mammalian species, or by detection of oocysts, or by recovery from rodents by bioassay. The possibility of transmission of T. gondii to swine via consumption of rodents, feed, and soil was confirmed.

Risk factors for transmission of Toxoplasma gondii on swine farms in Illinois.

Two epidemiologic studies of risk factors for transmission of Toxoplasma gondii to swine were conducted for farms in Illinois. The first study was a cross-sectional survey of swine farms from the state of Illinois pseudorabies testing program, in which farm owners or managers were interviewed by telephone regarding presence of risk factors for transmission of T. gondii on the farm. There were 123 farms surveyed that provided blood samples for at least 30 sows. The mean sow seroprevalence was 19.5% (median = 10.0%). Multiple regression analysis of the association of sow seroprevalence with outdoor housing of sows, cat access to sow areas, number of sows, open feed storage and water delivery, delayed removal of carcasses, and presence of rodents on the farm indicated that higher sow seroprevalence was associated with cat access to sows (P = 0.009) and fewer sows in the herd (P = 0.05). The second study was a field investigation of 47 swine farms (37 from the cross-sectional study). Data collection included obtaining blood samples from swine, cats, and rodents, and fecal samples from cats, heart and brain tissue from rodents, and feed, water, and soil samples for T. gondii examination. The risk of T. gondii transmission from cats and rodents to sows and finishing pigs was evaluated, taking into account housing conditions and herd size. Multiple regression analysis indicated that T. gondii seroprevalence in finishing pigs increased with more seropositive juvenile cats on the farm (P < 0.0001) and higher seroprevalence in house mice (P = 0.0023).(ABSTRACT TRUNCATED AT 250 WORDS)