Use of exposure databases for status and trends analysis. Exposure databases are useful for monitoring status and trends in environmental health. However, other supporting data are usually needed to infer human exposure or internal dose. Program planning and evaluation, environmental health surveillance, epidemiologic research, and contributions to international efforts are four major purposes for monitoring environmental exposure status and trends. Although databases play an important role in monitoring human exposure, certain methodological problems need to be overcome. The work group developed six criteria for meeting information needs for human exposure assessment. Areas that need attention are (1) specification of location, (2) specification of facility and chemical identifiers, (3) documentation of special populations at risk, (4) provision of early warning of new problems, (5) monitoring changes over time, and (6) enhancement of documentation. We tested these criteria by examining six available databases that might be used for monitoring exposure to contaminants in drinking water. Available data fell short of information needs. We drew four conclusions and offered several recommendations for each. First, available data systems lack adequate measures of human exposure. Second, data for monitoring exposures for many important population subgroups and environmental settings are inadequate. Third, an "early warning" system that monitors human exposures is needed. Fourth, designers of data-collection systems should consider the needs of users who monitor status and trends of human exposure.


Clinical associations of IgG antibodies to the ribonucleoprotein p67 polypeptide in patients with systemic lupus erythematosus. The ribonucleoprotein (RNP) p67 antigen was purified from rabbit thymus and used in an enzyme linked immunosorbent assay (ELISA) with low interassay variability to detect IgG antibodies to p67 in patients with autoimmune connective tissue diseases. These antibodies were found in eight (80%) patients with a clinical diagnosis of mixed connective tissue disease (MCTD) but also in 27 (40%) patients with systemic lupus erythematosus (SLE). Sixty six per cent of the 12 patients with SLE with high levels of antibodies to p67 (> 50 U) had three or more features of MCTD, including myositis, fibrosing alveolitis, Raynaud's phenomenon, and sclerodactyly. Antibodies to the p67 RNP were not associated with the presence or absence of renal disease in the patients with SLE. This study suggests that antibodies against the p67 RNP are markers for clinical features of MCTD even in the context of SLE.


Work related upper limb disorder: the relationship between pain, cumulative load, disability, and psychological factors. Repetitive strain injury, or work related upper limb disorder, provides an interesting paradigm for the study of the relative contribution of physical and psychological factors to the resulting pain and disability. Sixty three subjects were studied, comprising the work-force of a subsection of a large local industrial company, in whom pain in the arm related to work was known to be common. Ergonomic data were obtained by estimating the cumulative daily load on the wrist joint for each of four identified tasks. Data on the occurrence of pain, treatment sought, and disability were obtained by a structured self administered questionnaire. Psychological data were obtained by administering the Hospital Anxiety and Depression (HAD) scale, a self reported measure of anxiety and depression, and the Bradford Somatic Inventory (BSI), an inventory of somatic symptoms associated with anxiety and depression. The employment specific period prevalence of work related upper limb disorder was 81%, with 30% of the subjects having pain at the time of the study. Domestic disability was minimal in all but two subjects, though the use of devices such as jar openers at home was common (12 of 51 subjects). Medical advice was seldom sought. Twenty per cent of subjects had received anti-inflammatory drugs, 10% had received physiotherapy, and 47% had wrist splints. Pain was related to the tasks with the highest estimated daily loads, but a history of pain and current pain were associated with higher scores on the HAD and BSI scales, suggesting an interaction between physical and psychological factors.


Continuation of long term treatment with hydroxychloroquine in systemic lupus erythematosus and rheumatoid arthritis. Hydroxychloroquine is used for the treatment of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). Long term studies have shown a high rate of termination of hydroxychloroquine treatment in patients with RA. Although it has been shown that discontinuation of treatment with hydroxychloroquine is associated with exacerbation of SLE, long term maintenance rates of treatment with hydroxychloroquine in patients with SLE have not been investigated. Hydroxychloroquine use in patients with RA and SLE in a group of patients in a single community rheumatology practice was studied. Information was drawn from a computer drug use database containing details of the beginning and end of treatment. Data were analysed using life table methods. Four hundred and three treatment episodes (366 patients with RA, 37 patients with SLE) were observed over eight years. In patients with RA, the cumulative probability of discontinuing treatment was 37% at 12 months and 54% at 24 months. In contrast, hydroxychloroquine treatment of patients with SLE continued over significantly longer periods of time (p < 0.001); the discontinuation probabilities at 12 and 24 months were 8 and 24% respectively. Treatment terminations were predominantly for inefficacy; terminations for toxicity were limited to the first 19 months of treatment. No ocular toxicity was observed. Treatment of patients with RA in a community rheumatology practice with hydroxychloroquine has a low probability of long term continuation, mostly because of inadequate control of disease manifestations rather than toxicity. In patients with SLE, treatment with hydroxychloroquine has a significantly higher probability of long term continuation.


Methotrexate treatment of rheumatoid arthritis: is a fortnightly maintenance schedule enough? In 15 patients with rheumatoid arthritis who were in clinical remission the weekly regimen of methotrexate treatment was changed to fortnightly without a change in dose. A total of 13 patients completed a 12 month trial. No change in clinical or laboratory parameters occurred. There was no change in the use of analgesics or non-steroidal anti-inflammatory drugs and the patients remained in remission. Two patients had to be withdrawn after two and four months respectively because of a flare in disease activity. It is suggested that in most patients with rheumatoid arthritis who are treated with methotrexate and whose disease activity is stable a fortnightly regimen can be permitted without affecting drug efficacy.


Long term prognosis in yersinia arthritis: clinical and serological findings. Twenty two patients were followed up for a mean of 10.7 years after acute yersinia arthritis. Their clinical course, agglutination antibodies, antibodies against plasmid coded yersinia proteins, and laboratory parameters of inflammation were analysed. The clinical findings were unremarkable. Serum agglutinating antibodies against Yersinia enterocolitica were not found in any patient. Ten patients had no antibodies against plasmid coded bacterial proteins. Five patients showed IgG antibodies, one patient had an IgA and IgG result of questionable significance, and six patients had IgA and IgG antibodies in the immunoblot reaction. No evidence of systemic inflammation was found. The persistence of IgA or IgG antibodies, or both, did not have a discernible influence on the clinical course. The development of erosive changes in the sacroiliac joints occurred independently of yersinia infection.


Giant cell tumours in mandible and spine: a rare complication of Paget's disease of bone. The case of a man who was diagnosed as having polyostotic Paget's disease at the age of 52 years is described. He developed a rare neoplastic complication of Paget's disease--a giant cell tumour in his mandible, which was excised. Nine years after the diagnosis of this tumour he developed a new giant cell tumour arising from the L3 vertebral body. He was born in Avellino in Italy, from where five other cases of giant cell tumours arising from Pagetic bone disease have been reported. No family relationship between our patient and the other cases was established. His Paget's disease was particularly aggressive and resistant to treatment with two single high dose infusions of pamidronate almost two years apart.


The role of polymorphonuclear neutrophils in myocardial damage during ischemia and reperfusion. In addition to their traditional role in infection elimination and scavenging dead tissue, neutrophils may be injurious to myocardial tissues. Besides mechanical participation they produce toxic oxygen species, PAF, arachidonic acid metabolites, and release the proteases from intracellular granules. The deleterious role of PMN within myocardial ischemic focus depends also on the cooperation with platelets, macrophages, endothelial and mast cells. Rapid entry of neutrophils during ischemia contribute to arrhythmia, loss of coronary vasomotion, no-reflow phenomenon and extension of cellular injury. Further work is needed to define the precise role that neutrophils play in tissue injury and to propose an appropriate strategy by which PMN function can be modified to limit infarct size.


Chemiluminescence of bronchoalveolar macrophages: effect of adherence to plastic cuvette. The chemiluminescence of alveolar macrophages was determined in presence of luminol or lucigenin, either before or after the macrophage isolation by adherence to plastic. In presence of luminol, the purified and non-purified macrophages behave almost in the same way, and their chemiluminescence activity is stimulated both by zymosan and phorbol ester. On the other hand, the lucigenin chemiluminescence of non-purified macrophages is considerably higher than that obtained in presence of luminol, but is almost completely insensitive to the zymosan or phorbol ester stimulation. Moreover, the adherence purification considerably inhibits the lucigenin chemiluminescence. These results can be indicative of functionally different macrophage states.


Inhaled natural human interferon alpha induces bronchospastic reactions in asthmatics. There is a relationship between viral infection and aggravation of bronchial asthma symptoms. Viruses belong to the strongest and fast acting inducers of interferon (IFN), which, in turn, may enhance histamine release from effector cells. We have administered IFN intrabronchially to 49 patients suffering from bronchial asthma. The impairment of lung ventilation with significant decrease of forced expiratory volume in one second (FEV1) have been observed in majority of the patients who inhaled crude or purified IFN preparations.


IgG subclasses distribution in immunoglobulin preparations for intravenous use. The levels of IgG subclasses were determined, by radial immunodiffusion using monoclonal antibodies, in 16 production batches of two human immunoglobulin preparations for intravenous use--Immunovenin and Immunovenin-intact. While the partially degraded preparation--Immunovenin contained exclusively IgG1, the subclass distribution in Immunovenin-intact was close to that observed in normal human serum.


Association of giant multinucleated cells with deterioration of osteoinductive potency of bone matrix. A thirty four intramuscular implants of rat decalcified bone matrix into mice and rats were histologically inspected in order to find out if there is an inverse correlation between the induction of multinucleated giant cell formation and the bone induction activity. It was found that the presence of multinucleated giant cells which avidly resorbed the implanted matrix was almost always combined with the lack of local bone induction. In contrast, the specimens exhibiting intense bone induction were devoided of multinucleated giant cells. Our observation confirm the opinion expressed by Walters and Schneider that multinucleated giant cells induced by mineralized bone chips are not osteoclasts but a type of inflammatory cells, analogous if not identical to the "foreign body cells". It is postulated that formation of multinucleated giant cells is a morphological expression of hyperactivity of mononuclear phagocytes. Hyperactive multinucleated giant cells avidly resorb bone matrix and degrade bone inducing substance(s) like skeletal growth factor and/or bone morphogenetic peptides resulting in the deterioration or lack of bone induction.


Serological characterization of Proteus penneri species novum. Serological characterization of the first collection of the 20 Proteus penneri strains is presented. All anti-0 sera were examined in microagglutination, semi-quantitative precipitation and passive hemagglutination tests. Some P. penneri lipopolysaccharides showed strong cross-reactivity in passive hemagglutination additionally confirmed by inhibition in this test. Serological similarity between species within genus Proteus is discussed.


Antibacterial and antifungal agents. XV. Synthesis and antifungal activity of structural analogues of bifonazole and ketoconazole. The synthesis and antifungal activities of the cis- and trans-1-acetyl-4-[4-[[2-(1,1'-biphenyl-4-yl)-2-(1H-imidazol-1-ylmethy l)- 1,3-dioxolan-4-yl]-methoxy]phenyl)piperazines 3 and 4 are reported. Stereochemical assignments to diastereomeric pairs of cis/trans isomers were made on the basis of 1H- and 13C-NMR data. Among test derivatives the best activity was shown by the benzoyl esters of the cis- and trans-[2-(1,1'-biphenyl-4-yl)-2-(1H-imidazol-1-ylmethyl)-1,3-di oxolan-4- yl]methanols 9 and 10.


[Pharmacokinetics of hypolipedemic agents. 9. The question of biliary excretion of the hypolipidemic agent ciprofibrate (1) (2-(4-(2,2-dichlorocyclopropyl)-phenoxy)-2-methylpropionic acid]. After oral administration of 200 mg Ciprofibrate (1) to 10 patients, who were cholecystectomated with subsequent T-drainage, 1 could be determined in the collected bile only as its 1-O-beta-acylglucuronide to a very small amount. Approximative calculations have shown that two days after a single dose, less than 0.5% of the administered dose of 1 is excreted on the biliary route. It is an unsolved problem if there is an increase of biliary excretion after multiple dose.


[N-phosphorylated amino acids as potential cytostatics]. Phosphoramidic dichlorides 3 are synthesized by nucleophilic attack of methyl 2-(1-ethylenimino)-alkanoates 1 on POCl3 or by the reaction of amino acid methyl ester hydrochlorides 2 with POCl3. Subsequent treatment of 3 with ethylenimine affords the TEPA-analogs 4, and with 3-(1-ethylenimino)-1-propanol (6) the IFOSFAMID analogous 1,3,2-oxaza-phosphorines 5, the cytostatic activity of which was tested in the P388-leukemia of the CD2F1-mouse.


[Synthesis of enantiomerically pure 6,10-epoxybenzocycloocten-7-amines with CNS activity]. In an oxa-Pictet-Spengler reaction the methyl (S)-phenyllactate 6 and methyl levulinate (7a) are condensed to the 2-benzopyrans cis-8a and trans-8a, which react with CH3I to yield the dimethyl ethers cis-9a and trans-9a. Cis-9a and trans-9a can be separated by medium pressure liquid chromatography. In the subsequent Dieckmann-Cyclisation cis-9a is transformed to the laevorotatory beta-ketoester (-)-10a, while the dextrorotatory enantiomer (+)-10a is obtained from trans-9a after C-3-epimerisation. With Eu(hfc)3 the ketone (-)-11, prepared by saponification and decarboxylation of (-)-10a, proves to be enantiomerically pure. By reductive amination, ketone (-)-11 is transformed to the amines (-)-12a and (-)-12b. Symptoms typical for central damping are caused after i.p. application of (-)-12a and (-)-12b to mice. In the mouse writhing-test (-)-12a HCl affords an ED50-value of 7.0 mg/kg, comparable with the ED50-value of tramadol.


A call to the international nursing community: put AIDS in developing countries on the agenda. In some African countries 10%-15% of the middle class may due during the 1990s and at the present time the human infrastructure is being eroded by HIV. Over 90% of home and community care is being supervised by nursing and midwifery personnel in some developing countries. What will happen if these carers become ill or die? Peter Kerr provided a micro view of HIV disease in developing countries in 'Health care crisis in Africa', August ANJ. In this article he discusses the macro implications for developing countries as they encounter the consequences of what is likely to be the most devastating epidemic of this century.


Casemix is coming.

Casemix is coming. Casemix is the term used to describe the types of patients being treated in hospitals. During the 1980s, the main casemix classification adopted in Australia utilised diagnosis related groups (DRGs) and Australian Governments are now moving towards output-based funding of hospitals using DRGs. Changes to hospital funding mechanisms, increased focus on the output or product of hospital services and the availability of more information linking service provision to costs will have an impact on all nurses employed in hospital settings.


Experimental exposure of broiler chickens to boric acid to assess clinical signs and lesions of toxicosis. One-day-old broiler chicks were housed on litter treated with 0, 0.9, 3.6, or 7.2 kg of boric acid per 9.3 m2 of floor space. After 15 days, average body weights were 340.7 g, 278.1 g, and 213.2 g for chickens exposed to 0, 3.6, and 7.2 kg boric acid/9.3 m2, respectively (P < or = 0.05). Feed conversion was 1.68 and 2.29 in chickens exposed to 0 and 7.2 kg boric acid/9.3 m2 (P < or = 0.05). In a second experiment, after birds ingested feed treated with 2500 and 5000 ppm of boric acid for 2 weeks, body weights were 254.8 g and 149.6 g, respectively. The chickens in the control group weighed an average of 285.0 g (P < or = 0.05). The 5000 ppm treatment group had a feed conversion of 1.70, vs. 1.45 in the controls (P < or = 0.05). A dose-related feathering abnormality was evident in treated chickens. No significant histopathologic changes were observed. Livability, weight gain, and feed conversion were not adversely affected in broilers as a result of exposure to litter treated with boric acid at recommended levels of 0.4-0.9 kg/9.3 m2.


Polymerase chain reaction amplification for direct detection of chicken anemia virus DNA in tissues and sera. Direct detection of chicken anemia virus (CAV) DNA in tissues and sera was investigated by a polymerase chain reaction (PCR) assay. Using a pair of primers constructed to amplify the coding sequence of the CAV DNA genome, the PCR assay was shown to be extremely sensitive, being able to detect 1 fg of CAV replicative form DNA. The oligonucleotide primers used for the PCR yielded 583 base-pair (bp) amplified product, which was sized by ethidium bromide-agarose gel electrophoresis. Tissue samples from seven cases of suspected chicken infectious anemia were obtained for CAV isolation. DNA extracted from the homogenized suspension of pooled tissues of each case was analyzed by the PCR assay. Results showed that five of the seven cases were positive for CAV DNA by PCR, whereas CAV was isolated from four cases only. The PCR assay also detected CAV DNA in two of 37 serum samples from disease-free chickens. The specificity of PCR was confirmed by chemiluminescence dot-blot analysis of the amplified products.


Increasing incidence of anemia among clinically ill Georgia broilers: 1988-89 vs. 1990. The incidence of anemia in clinically ill Georgia broilers climbed from 66.4% (324/488) during 1988-89 to 80.9% (531/656) during 1990. The incidence of polycythemia fell from 1.6% (8/488) during 1988-89 to 1.5% (10/656) during 1990. Specifically, compared with 1988-89, the 1990 incidence of anemia increased significantly in chicks at age 7 days (P = 0.0002) and 28 days (P = 0.05). We have no certain explanation for this shifting incidence of anemia in clinically ill Georgia broilers. Anemic chicks have plasma that contains virus particles with morphologic characteristics consistent with a virus (chicken anemia agent [CAA]) known to cause anemia in chickens. If CAA is the predominant etiology for anemia in clinically ill Georgia broilers, then our observation could be easily explained. The increasing rate of anemia could indicate a decline in broiler health over time.


Effect of age of maternal flock and strain on the incidence of tibial dyschondroplasia in growing chicks. Two experiments were conducted to study the effects of age of the maternal flock (27 vs. 57 weeks of age at time the eggs were set) and strain of broiler chicks (Cobb 500, Ross, Arbor Acres, and Avian) on the incidence of tibial dyschondroplasia (TD) in broiler cockerels grown to 3 weeks of age. The chicks were fed a corn/soybean meal-based diet containing 3145 kcal metabolizable energy corrected for nitrogen retention (MEn)/kg, 24% protein, and 0.50% available phosphorus. Each treatment was replicated four times with eight chicks per pen. In Expt. 1, chicks from the old parent flock had significantly higher body weights (P < or = 0.05) than those from the young parent flock. There were no significant differences in feed efficiency or incidence of TD. In Expt. 2, body weights, feed efficiency, and TD were unaffected by strain.


Comparison of MTT colorimetric assay and tritiated thymidine uptake for lymphocyte proliferation assays using chicken splenocytes. The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) colorimetric assay was compared with the conventional tritiated thymidine deoxyriboside (3H-TdR) incorporation for assay of lymphocyte blastogenesis using mononuclear cells isolated from the spleens of specific-pathogen-free chickens. The study was undertaken in an effort to simplify methods for assessing avian lymphocyte proliferation, specifically for evaluating response to mitogens or for indirect measurement of T-cell growth factors. The results from stimulated cells in both assay methods were significantly different from results from the control cells, and the MTT assay results regressed in a significant linear manner on counts from 3H-TdR incorporation. On this basis, the MTT assay is a valid test for evaluation of lymphocyte proliferation of chicken splenocytes.


Failure of the Congo red dye uptake test to discriminate between virulent and avirulent avian Escherichia coli. Twenty avian Escherichia coli isolates from normal and diseased chickens were compared by use of three virulence tests. These tests included the uptake of Congo red dye, an embryo lethality test, and a quantitative microtiter complement resistance test. A direct correlation was seen between the results of the complement resistance test and the embryo lethality test. The results of the Congo red test did not correlate with the two other tests.


A feather-trap system for the removal of chicken feathers from laboratory sewage. A simple feather-trap system is described for use on the drain lines of buildings housing poultry for research or other purposes where floors are frequently washed. The trap uses disposable plastic-mesh bags that can efficiently remove almost all feathers from the water, preventing sewer lines from being blocked by compacted feathers. Critical measurements and operational procedures are described.


Immunization of young chickens by trickle infection with Eimeria tenella. Immunization of chickens was attempted with low levels of Eimeria tenella oocysts (50 oocysts per day) over the first 1 or 2 weeks of life--the "trickle infection" (TI) method. When chickens were immunized by TI at 0-13 days of age, no cecal lesions and a reduced number of oocysts in ceca were observed after challenge at 17 days of age. TI at 0-6 days of age conferred better protection against challenge with E. tenella than did TI at 7-13 days. However, cecal lesions were observed in almost all of these chickens. These findings indicated that TI for 2 weeks (0-13 days of age) provided better immunity than TI for 1 week (0-6 or 7-13 days of age).


Studies on the effect of concurrent infections of Ascaridia dissimilis and Eimeria meleagrimitis in turkeys. The effects of concurrent infections of Ascaridia dissimilis and Eimeria meleagrimitis in turkeys were studied in two separate trials. In the first trial, newly larvated ova were used to inoculate poults 7 or 3 days before, on the same day as, or 3 days after the poults received E. meleagrimitis. Poults receiving the A. dissimilis 3 days before, on the same day as, or 3 days after receiving E. meleagrimitis had significantly lower total oocyst production than the E. meleagrimitis-positive control. In the second trial, larvated ova that were approximately 100 days old were used in the same regimen. In this trial, poults that were inoculated with A. dissimilis 3 days before or 3 days after receiving the E. meleagrimitis produced significantly fewer oocysts than poults inoculated simultaneously with both parasites. Poults inoculated with A. dissimilis 3 days before receiving E. meleagrimitis also had significantly fewer third-stage nematodes than the A. dissimilis-positive controls. There were no significant differences in weight gain between treatments in either trial.


Simulation of maternal immunity by inoculation of immune yolk preparations into the yolk sac of 1-day-old chickens. Yolk harvested from eggs laid by hens hyperimmunized with killed Newcastle disease virus (NDV) was inoculated into the yolk sac of 1-day-old specific-pathogen-free (SPF) chickens. Serum hemagglutination-inhibition antibody titers reached maximum levels 1 to 4 days after yolk inoculation and declined at a rate similar to that reported for naturally acquired maternal antibody. Expected levels of immune interference were observed when yolk-inoculated chickens were vaccinated with a conventional oil-emulsion NDV vaccine. These results show that yolk-sac inoculation with yolk antibody is a suitable approach for producing maternally immune chickens for laboratory studies.


An inexpensive system to monitor air flow in isolation units. Isolation units are used extensively for conducting infectious disease research in poultry. By necessity, these units are airtight and receive air only through electrically powered ventilation systems. Therefore, interruptions in electrical service to these units present a serious hazard to the animals they contain. A system was designed to monitor the air flow through isolation units and to alert animal caretakers in the event of any interruption in air flow. The "intelligence" of the system relies on an electronic monitor connected to a telephone line that places alerting telephone calls when it detects loss of air flow to the units. The system is constructed from easily acquired and relatively inexpensive parts and components.


Comparisons of packed cell volumes (PCVs) from so-called chicken anemia agent (CAA; a virus)-free broilers to PCVs from CAA-free specific-pathogen-free leghorns. Packed cell volumes (PCVs) from 3-, 7-, 14-, 21-, 28-, and 35-day-old clinically healthy chicken anemia agent (CAA)-free and specific-pathogen-free (SPF) leghorn chicks were compared with PCVs from age-matched clinically healthy CAA-free broiler chicks. The PCVs of the SPF chicks regressed significantly (F = 20.6, df = 2/3, P < 0.025) on age in a linear fashion. The PCVs of the broiler chicks regressed significantly (F = 9.56, df = 2/3, P < 0.05) on age in a cubic parabola. The mean PCVs of the broiler chicks were significantly (P < 0.05) higher than PCVs of SPF chicks at every tested time interval. Results indicate that PCV values are higher in broiler chicks than in SPF leghorn chicks, and that PCVs increase as chicks age. Clinicians, diagnosticians, and investigators who intend to work with chicken blood must be aware of these differences.


Effect of hypoxia and diet on spontaneous turkey cardiomyopathy (round-heart disease). The purpose of this study was to examine the effect of hypoxia-induced hypoxemia and rate of growth on spontaneous dilatory cardiomyopathy in turkey poults. Turkey poults grown in a hypobaric chamber at an atmospheric pressure of 592 mmHg (calculated partial pressure of oxygen: 124 mmHg; calculated altitude and O2 equivalents: 2054 m and 16.3%) on a rapid-growth diet developed a mainly right ventricular dilatory cardiomyopathy typical of the acute form of spontaneous turkey cardiomyopathy (STC). Poults grown in a hypobaric chamber on a slow-growth diet had a much lower incidence of STC. Control poults at atmospheric pressure, altitude 295 m (calculated atmospheric pressure: 735 mmHg; partial pressure of oxygen: 154 mmHg, 20.26% O2), on a rapid-growth diet had an incidence of STC similar to that of the slow-growth hypobaric group, while those on a slow-growth diet had a lower incidence.


Plasma bile acid concentrations in response to feeding in peregrine falcons (Falco peregrinus). A significant post-prandial increase of plasma bile acid concentration (PBAC) was observed in peregrine falcons (Falco peregrinus). In order to avoid physiological food-induced elevations in PBAC, which can complicate interpretation of plasma chemistry results in birds of prey, it is recommended that blood samples be obtained after a fasting period of at least 24 hours.


Sarcocystis in mallards on the Southern High Plains of Texas. Two hundred thirty-one mallards (Anas platyrhynchos) collected during 1989-91 on the Southern High Plains of Texas were examined for macroscopically detectable Sarcocystis spp. Only eight adult mallards were infected. Based on small quantified tissue samples, five, one, and two birds had light, medium, and heavy infections, respectively. Prevalences were 4%, 8%, 9%, and 0% across two consecutive summer and winter periods, respectively. A new locality record for Sarcocystis spp. was established in both migratory and breeding populations of mallards. Our data suggest that geographic regions other than the Southern High Plains are more important in the transmission of Sarcocystis spp. in mallards.


Pullorum disease in Delaware roasters.

Pullorum disease in Delaware roasters. An index case of pullorum disease in 4-week-old roasters from a completely integrated poultry operation is reported from Delaware. Severe articular and periarticular swelling of hock and/or wing joints was observed on postmortem examination. Also seen were moderate to severe hydropericardium and large white-gray nodules in the heart and gizzard that grossly could have been taken for tumors. The livers showed multiple small white foci, petechial hemorrhages, and swelling. Initially, Salmonella pullorum was isolated from the articular fluid by direct culture on MacConkey agar, but it was not isolated from liver, spleen, or heart samples after enrichment in selenite broth.


A syndrome in commercial turkeys in California and Oregon characterized by a rear-limb necrotizing skeletal myopathy. A necrotizing skeletal myopathy of rear limbs was diagnosed in 17 flocks of commercial turkeys. The mean mortality attributed to the myopathy was 2.29% (range = 0.13-9.7%) over a mean period of 9.6 days (range = 6-14 days). The mean age of the birds at the time of onset was 7.4 weeks (range = 4-10.5 weeks). Clinically, birds experienced an episode of watery droppings and high-pitched crying, followed by rear-limb paresis or paralysis. Creatine kinase and aspartate aminotransferase were markedly elevated in birds with the myopathy. Grossly, a few birds had pale streaking in the muscles of the thighs and legs. Histologically, acute and subacute degeneration was present in myofibers of the legs, abdomen, thighs, back, and tail. The subacute lesion was characterized by marked sarcolemmal cell proliferation. Feed analyses ruled out selenium deficiency and the presence of mycotoxins as etiologies. Monensin was present in approved usage or only slightly elevated levels. A known potentiating antibiotic was being used concurrently with monensin in only one flock.


Pyogranulomatous typhlitis and hepatitis of market turkeys. Pyogranulomatous lesions in the liver and cecum were observed in groups of 7-to-8-week-old market turkeys. Necropsy revealed multifocal lesions in the liver and the presence of cecal cores that, in some cases, distended the cecum to the point of rupture. Mortality was associated with the ruptured ceca. Two successive flocks also succumbed to these lesions. An epizootiological study was performed to determine the cause and/or initiation of lesions leading to the formation of cecal cores.


Toxicosis in broiler chicks due to excess dietary ethoxyquin. A commercial broiler company experienced a sudden increase of mortality in four flocks at two separate locations. Affected houses had red dust on inside walls and on the equipment. Affected chickens were 10 to 18 days old. Grossly, chickens had pale, swollen kidneys, dark-brown enlarged livers, and urates in joint spaces. Histologically, the kidneys had multifocal proximal tubular necrosis, the livers had dilated sinusoids, biliary hyperplasia, and accumulation of brown pigment in hepatocytes and bile ducts. One starter ration was analyzed, and it contained 6500 mg ethoxyquin/kg. The red dust in the houses and the brown pigment in livers were interpreted to be ethoxyquin. Clinical signs and histologic lesions were reproduced experimentally with 12,500 mg ethoxyquin/kg feed.


Aeromonas hydrophila conjunctivitis in a pet parrot (Amazona versicolor). A bilateral conjunctivitis in a pet parrot (Amazona versicolor) is reported. Aeromonas hydrophila was isolated in pure culture from both eyes and considered of diagnostic significance. Biochemical characteristics and antimicrobial susceptibility of the strain were studied, as were the factors that could have contributed to the clinical conjunctivitis and the role of A. hydrophila as an opportunistic pathogen.


Sarcoid granuloma-like lesions in the lungs of broiler chicks. Sarcoid granuloma-like lesions were observed in the lungs of two broiler chicks showing depressed growth. Multinucleated giant cells in the lesions often contained foreign bodies comprising amorphous and laminated material. These were morphologically similar to Schaumann bodies found in human sarcoidosis. This is the first report of sarcoid granuloma-like lesions and Schaumann body-like material in chickens.


Tracheal aspergillosis in 6 1/2-week-old chickens caused by Aspergillus flavus. A case of localized tracheal aspergillosis in 6 1/2-week-old single-comb white leghorn pullets caused by Aspergillus flavus is documented. Yellow caseous plaques adherent to the mucosal surface of the tracheas were observed grossly. In several tracheas, the plaques occluded the lumina, and the surrounding tracheal walls were reddened. Histologically, the mucosa was necrotic and infiltrated with macrophages, and fibroplasia was evident in the subadjacent tracheal wall. The lumen of the trachea was almost completely occluded by a combination of fungal mycelia and pyogranulomatous exudate. Portions of tracheal cartilage were elevated into the lumen of the trachea. Other than a sudden increase in mortality to 0.5% per day, there was no evidence of disease in the flock. Depletion of bursal lymphocytes, with concomitant cryptosporidiosis, was evident on histological examination. Acute infectious bursal disease was diagnosed in the succeeding flock at this ranch based upon serology and typical histology.


Bacteremia and vegetative endocarditis associated with a heart murmur in a blue-and-gold macaw. A 6-year-old male blue-and-gold macaw (Ara ararauna) was presented with severe weakness, anorexia, and weight loss of 2 weeks duration. Cardiac auscultation revealed a soft systolic murmur. Blood cultures collected both antemortem and postmortem yielded pure isolates of Enterobacter cloacae. At necropsy, vegetative endocarditis was found involving the left atrioventricular valve. Microscopically, the lesion on the valve was characterized by a mixture of necrotic material, colonies of gram-negative bacteria, fibrosis, and inflammatory infiltrate consisting primarily of heterophils.


Myocarditis as a component of psittacine proventricular dilatation syndrome in a Patagonian conure. Psittacine proventricular dilatation syndrome was diagnosed at necropsy in a Patagonian conure. Gross and histopathological lesions in the proventriculus, ventriculus, and brain were similar to those previously reported. In addition, severe myocarditis was a prominent feature of this case, a finding not previously defined as a significant component of the condition.


Protection of chickens from Newcastle and Marek's diseases with a recombinant herpesvirus of turkeys vaccine expressing the Newcastle disease virus fusion protein. Recombinant strains of herpesvirus of turkeys (HVT) were constructed that contain either the fusion protein gene or the hemagglutinin-neuraminidase gene of Newcastle disease virus (NDV) inserted into a nonessential gene of HVT. Expression of the NDV antigens was regulated from a strong promoter element derived from the Rous sarcoma virus long terminal repeat. Recombinant HVT strains were stable and fully infectious in cell culture and in chickens. Chickens receiving a single intra-abdominal inoculation at 1 day of age with recombinant HVT expressing the NDV fusion protein had an immunological response and were protected (> 90%) against lethal intramuscular challenge at 28 days of age with the neurotropic velogenic NDV strain Texas GB. Recombinant HVT expressing the NDV hemagglutinin-neuraminidase provided partial protection (47%) against the same challenge. Chickens vaccinated with recombinant HVT vaccines had low levels of protection against NDV replication in the trachea when challenged ocularly. Recombinant HVT vaccines and the parent HVT strain provided similar levels of protection to chickens challenged with the very virulent RB1B strain of Marek's disease virus, indicating that insertion of foreign sequences into the HVT genome did not compromise the ability of HVT to protect against Marek's disease.


Effect of used litter from floor pens of adult broilers on Salmonella colonization of broiler chicks. The effect of used pine-shaving litter from broiler floor pens on Salmonella colonization resistance was evaluated in broiler chicks. One-day-old chicks were placed in floor pens on fresh unused litter or on used litter. All chicks were challenged orally with 10(4) S. typhimurium at 3 days of age. The study was replicated in three trials with used litter that was collected and stored for 1 day (Trial 1), 4 days (Trial 2), or 50 days (Trial 3) before the start of each trial. Cecal concentrations of volatile fatty acids (VFAs) were significantly higher (P < 0.05) in chicks placed on used litter than in chicks on new litter. In all three trials, the number of Salmonella in the cecal contents and the number of Salmonella cecal-culture-positive chicks was significantly lower (P < 0.01) at 10 days and 20 days of age in the chicks on used litter than in the chicks on new litter. The results indicated that newly hatched chicks reared on used litter had higher cecal VFA concentrations and higher resistance to Salmonella colonization than chicks reared on new litter.


Experimental Streptococcus bovis infections in pigeons. Thirty pigeons were experimentally infected with Streptococcus bovis using an intravenous infection model. Ninety percent of the inoculated pigeons developed clinical disease. Disease signs included acute death, inability to fly, lameness, inappetence, emaciation, polyuria, and the production of slimy, green droppings. At necropsy, the septicemic character of the disease was evident. Typical lesions included extensive well-circumscribed areas of necrosis in the pectoral muscle, tenosynovitis of the tendon of the Musculus pectoralis profundus, and arthritis of the stifle, tibiotarsal, or shoulder joints. Focal myocardial necrosis also was seen. Meningitis and encephalitis occurred in the cerebrum and the cerebellum. Disease signs and lesions described here after experimental infection were similar to those in naturally occurring cases of S. bovis septicemia.


Effect of virginiamycin on serum carotenoid levels and long, segmented, filamentous organisms in broiler chicks. Virginiamycin (Stafac 20) was mixed with feed at three levels recommended for chickens--5.5, 11, and 22 mg/kg (respectively 5, 10 and 20 g/ton)--and fed to broiler chicks. When fed from 1 day through 2 weeks of age, the drug appeared to retard infection of the lower small intestine by long, segmented, filamentous organisms (LSFOs), and at 2 weeks of age serum carotenoids in treated chicks were significantly higher than levels found in unmedicated chicks. However, as chicks were grown out to 4 and 6 weeks of age, the drug did not completely prevent eventual LSFO infection, even at the highest dose, and mean serum carotenoid levels in treated chicks were not significantly different from levels in control chicks. However, chicks fed at the level of 22 mg/kg had fewer LSFOs. Withdrawal of virginiamycin from treated chicks at 4 weeks of age allowed LSFO infection to occur but did not significantly affect serum carotenoid levels. When the drug was fed for 2 weeks at 22 mg/kg to 2-week-old chicks already infected with LSFOs, the bacteria could no longer be detected, suggesting that virginiamycin may aid the natural decline in LSFO population. Coincident with this treatment, serum carotenoids were higher, but not significantly so. Virginiamycin did not significantly increase the mean body weights of chicks in any of these experiments.


Evaluation of the kidney as a potential site of avian influenza virus persistence in chickens. One-day-old chickens were inoculated intravenously with one of three low-pathogenicity avian-origin influenza isolates. On day 5 postinoculation (PI), the frequency of influenza virus isolation from cloacal swabs following challenge with each isolate ranged from 83% to 100% for clinically normal euthanatized chickens. Influenza virus was also frequently isolated from kidneys of these chickens (47%) and from chickens that died (100%). Kidneys positive for virus isolation had lesions of nephrosis and/or acute nephritis, and influenza viral nucleoprotein was demonstrated in nuclei and cytoplasm of necrotic renal tubule epithelium. On sampling days 28 and 45/60 PI, influenza virus was neither isolated from nor immunohistochemically demonstrated in kidneys (0/125); however, the kidneys (47%) did have chronic histologic lesions that suggested previous influenza virus infection of the kidneys. Influenza virus was isolated from cloacal swabs of two of 44 chickens on day 28 PI, but all cloacal swabs were negative for virus recovery on sampling day 45/60 PI (0/81). These results indicate that replication of influenza virus in renal tubule epithelial cells did not result in persistence of type A influenza virus in this immunologically privileged site.


Toxin production by Clostridium perfringens isolated from broiler chickens and capercaillies (Tetrao urogallus) with and without necrotizing enteritis. A total of 192 isolates of Clostridium perfringens were isolated from 99 broiler chickens and 93 capercaillies (Tetrao urogallus). Fifty of the isolates from broilers and 44 of the isolates from capercaillies were from birds with necrotizing enteritis, and the remainder were from birds without this disease. The isolates were tested for the production of three major toxins (alpha, beta, and epsilon) and four minor toxins (theta, gelatinase, mu, and nu). All isolates were found to be C. perfringens type A. Alpha toxin was produced in significantly larger amounts by isolates from birds with necrotizing enteritis than by isolates from birds without the disease, regardless of bird species. Isolates from broilers produced significantly more alpha toxin than did isolates from capercaillies.


Pathologic and serologic characterization of a variant of duck hepatitis type I virus. A duck hepatitis virus (DHV), isolated from ducks on a farm in Virginia in 1963, was shown to be only partially related to DHV type I (DHV-I) in cross-neutralization and in cross-protection tests. The virus, named DHV-Ia, apparently is a serologic variant of DHV-I; both viruses are serologically distinct from DHV type III. Pathologic responses to DHV-Ia were similar to those described for DHV-I infection.


Optimum conditions for the chicken lymphocyte transformation test. Optimum conditions for chicken (Gallus gallus) lymphocyte transformation tests were determined. Thrice-washed chicken buffy-coat cells obtained after slow centrifugation (40 x g for 10 minutes) responded substantially better to mitogenic stimulation than lymphocytes isolated on separation media containing Ficoll. Maximum responses were obtained with 2 x 10(7) lymphoid cells/ml. Responses to the mitogens were greatest when fetal bovine serum was used at a 5% concentration or pooled chicken serum and autologous plasma were used at a 1.25% concentration. Optimum mitogen concentrations varied with individual birds, timing of the culture, temperature of incubation, and serum concentration in the cultures. When 1.25% chicken serum was used in the cultures, responses were usually greatest with final concentrations of 30-50 micrograms/ml of concanavalin A (Con A) and 30-50 micrograms/ml of phytohemagglutinin-P (PHA-P). The optimum concentration of pokeweed mitogen (PWM) varied from 1 to 40 micrograms/ml among the birds and was practically impossible to establish in general. The incubation in humidified air with 5% CO2 was significantly better at 40 C than at 37 C. The total culture time of 40 hours including pulsing with 3H-thymidine during the final 16 hours of incubation was the best for Con A- and PHA-P-stimulated cells, whereas a longer incubation of 64 hours gave the highest results with PWM stimulations.


Nephrotropic properties demonstrated by A/chicken/Alabama/75 (H4N8) following intravenous challenge of chickens. Tissue tropism properties of A/chicken/Alabama/75 (H4N8) were examined after intravenous inoculation of 5-week-old specific-pathogen-free chickens. From 14 clinically normal chickens euthanatized on days 1-20 postinoculation, the frequencies of virus recovery were highest for cloacal swabs (86%), bursal swabs (64%), and kidney tissues (64%) and lowest for tracheal swabs (14%), thymus tissues (14%), bone-marrow swabs (7%), and brain tissues (0%). Evidence that the high frequency of virus recovery from kidney tissues was associated with virus replication in the kidney tissues was provided by high virus titers, ranging up to 10(9.5) mean embryo infectious dose per gram of kidney tissue, and by identification of intranuclear and intracytoplasmic type A influenza nucleoprotein in kidney cells using immunohistochemistry. Virus-recovery and virus titer results from three chickens that died on days 4 and 5 postinoculation paralleled the results from the clinically normal chickens. These findings indicate that A/chicken/Alabama/75 has nephrotropic properties similar to nephrotropic properties previously reported for waterfowl-origin type A influenza viruses and provide evidence that kidney lesions could be manifestations of systemic influenza infections in commercial laying chickens.


Mycoplasma gallisepticum vaccination-challenge: an egg-production model. Specific-pathogen-free layer hens in maximum lay were exposed by aerosol to a broth culture of Mycoplasma gallisepticum R' strain. Egg-production loss of greater than 50% was evident 7-14 days following challenge of unvaccinated chickens, with a gradual recovery during the next several days. Various vaccine preparations were tested to determine the effect in the model. All vaccinated chickens exhibited significantly (P < or = 0.05) lower egg-production loss than the unvaccinated controls. The model provides a method for testing treatment effects on egg-production losses resulting from controlled exposure to M. gallisepticum and may be useful in simulating field exposure.


Turkey macrophage and heterophil bactericidal activity against Pasteurella multocida. Bactericidal activity of turkey macrophages and heterophils was demonstrated in an in vitro colorimetric bactericidal assay. Two vaccine strains and one field isolate of Pasteurella multocida A:3,4 and a single isolate each of Escherichia coli and Staphylococcus aureus were compared for susceptibility to the bactericidal activity of turkey macrophages and heterophils. Only P. multocida A:3,4-strain M-9 (the least virulent strain) was susceptible to macrophage bactericidal activity in the absence of specific immune serum, whereas all three P. multocida A:3,4 organisms were killed when opsonized with specific immune serum. E. coli was susceptible to the bactericidal activity of macrophages, and S. aureus was resistant. All bacteria tested were highly sensitive to the bactericidal activity of intact turkey heterophils, regardless of the opsonin treatment. Electron microscopic findings suggested that heterophils may kill extracellular P. multocida. Only S. aureus and E. coli were killed by lysed heterophils.


Re-evaluation of the pathogenicity of A/chicken/Alabama/75 (H4N8) influenza virus. Avian influenza (AI) virus A/chicken/Alabama/7395/75 (H4N8), a putatively non-pathogenic virus associated with a self-limiting outbreak of severe disease in commercial layers, was selectively passed in chickens or in cell cultures and then in chickens to determine whether virus with increased pathogenicity would emerge. When 20 derivatives of the parental virus were each inoculated intranasally and intratracheally in leghorn hens, mortality rates ranged from zero (0/24) to 25% (6/24); mortality was 4% (1/24) for hens inoculated with the parental virus. Many virus reisolates (51/144) from hens that died exhibited high pathogenicity, killing at least six of eight intravenously inoculated 4-week-old chickens. Most derivatives examined produced plaques in trypsin-free cell cultures more efficiently than the parental virus, but the highest plaquing efficiencies observed (10%) were lower than would be expected (100%) for highly pathogenic subtype H5 or H7 AI viruses. These results confirm that the Alabama H4N8 virus can acquire increased pathogenicity upon passage in chickens and suggest that it may have acted alone in producing the severe disease observed in laying chickens in Alabama.


Significance of adaptation mechanisms in adriamycin induced congestive heart failure. Natural history of myocardial dysfunction due to chronic contractile deficit consists of physiological and pathophysiological adaptations culminating in congestive heart failure. Among the mechanisms considered is the combination of compensatory as well as the harmful overcompensatory role of the adrenergic system during the genesis of a congestive heart failure "spiral" due to the chronic treatment with adriamycin. Refractoriness of this spiral to various inotropic agents may involve reduced sympathetic support of the myocardium, structural loss of contractile elements and abnormalities of the Ca2+ metabolism.


Myocardial contractile protein ATPase activities in adrenalectomized and thyroidectomized rats. This report compares the effects of adrenalectomy and thyroidectomy, with and without hormone replacement, on loss of contractile protein ATPase activities. The rationale for this study was derived from the similarities in their intracellular receptors, mechanisms of action, and the large number of proteins regulated by both hormones. Rats were adrenalectomized, thyroidectomized, or both, and were subsequently treated for 6 weeks with hydrocortisone, triiodothyronine, or saline. Sham-operated rats were given saline for the same period of time. Six weeks of adrenal insufficiency resulted in diminished enzymatic activity of myofibrillar, Ca(2+)-activated myosin ATPase, and actin-activated myosin ATPase fractions. Treatment with hydrocortisone prevented the decline in enzymatic activity due to adrenalectomy. Likewise, thyroidectomy caused a loss of enzymatic activity which was prevented by treatment with triiodothyronine. The full deleterious effect of combined ablation could be partially prevented by treatment with either hydrocortisone or triiodothyronine, but the latter was most effective. The results suggest that hydrocortisone and triiodothyronine each had significant positive effects in the presence of the other, but not in its absence, on the activity of myofibrillar Ca(2+)-dependent Mg-ATPase and Ca(2+)-activated myosin ATPase. The effects of these two hormones on actin-activated myosin ATPase activity were more independent of each other. We conclude that the actions of thyroid and glucocorticoid hormones on the heart are interrelated and that optimum myocardial function results from their combined action.


Vaccination of turkeys with cell-free culture filtrate of Pasteurella multocida: effects of dilution, iron chelation, and heterologous challenge. Two experiments were done to further define cell-free culture filtrate (CCF) from Pasteurella multocida and its endotoxin content in protecting turkeys against challenge. In the first experiment, the greater-than-30,000-molecular-weight fraction of P. multocida strain R44/6 (serotype 3/4/9/12) CCF was used in 10-fold dilutions given by air-sac inoculation or aerosol to vaccinate turkeys, which were subsequently challenged with either homologous (P-1059, serotype 3) or heterologous (X-73, serotype 1) strains. Endotoxin content of the CCF fraction was high. Compared with positive controls given either live Clemson University vaccine or a commercial bacterin, homologous protection was provided by undiluted CCF and 1:10 dilutions of CCF, but there was no heterologous protection. In the second experiment, CCF of strain R44/6 in regular and iron-limiting media and CCF of strain FC127B (serotype 1/4) were used alone or in combination to vaccinate turkeys, which were challenged as in the first experiment. Homologous but not heterologous protection occurred, even though growth of strain R44/6 in iron-limiting media reduced endotoxin content of CCF by approximately 93%. These results indicate that endotoxin levels of less than 10% but greater than 1% of those in CCF from regular media are sufficient to induce protection in turkeys against homologous challenge but that CCF from either regular or iron-limiting medium does not provide protection against heterologous challenge.


Evaluation of the efficacy of an oil-emulsion bacterin for protecting chickens against Salmonella enteritidis. To assess the potential protective efficacy of a Salmonella enteritidis bacterin, an acetone-killed oil-emulsion vaccine was prepared from a phage type 13a S. enteritidis strain and administered subcutaneously to hens in two experiments. Hens were housed individually, and every other hen was vaccinated (at 23 weeks of age in one experiment and at 45 weeks in the other). A second (booster) bacterin injection was administered 6 weeks later in both experiments. Three weeks after the second vaccination, all hens were challenged with an oral dose of approximately 10(9) cells of a heterologous (phage type 14b) S. enteritidis strain. In both trials, S. enteritidis was isolated from fewer internal organs (spleens, ovaries, and oviducts) and pools of egg contents from vaccinated hens than from unvaccinated control hens. Vaccination did not, however, affect the percentage of hens that shed S. enteritidis in feces in either experiment.


Predicting functional recovery from ischemia in the rat myocardium. Depletion of high-energy phosphates, accumulation of inorganic phosphate and intracellular acidosis have each been proposed as important events in the transition from reversible to irreversible ischemic injury. To assess whether each variable is predictive of functional recovery on reperfusion, these were measured in the isolated isovolumic rat heart using 31P NMR. Perfused hearts were subjected to either 10, 12 or 40 min of normothermic ischemia followed by 40 min of reperfusion. Hearts were then freeze-clamped for further analysis of phosphate metabolites by NMR and ion chromatography. High-energy phosphates, Pi, phosphomonoesters and pH were measured by 31P NMR spectroscopy at 2 minute intervals. Heart rate and developed pressure were monitored simultaneously. All hearts undergoing 10 min of ischemia and 40% of hearts subjected to 12 min of ischemia demonstrated good functional recovery. The remainder of hearts ischemic for 12 min went into contracture on reperfusion with little return of function. Hearts subject to 40 min of ischemia went into ischemic contracture and showed no recovery on reperfusion. Intracellular pH, [ATP], and [Pi] measured prior to reperfusion did not predict the extent of recovery. However, phosphomonoesters were detected prior to reperfusion in all hearts that did not recover well, but were not observed in hearts that showed good mechanical recovery. Analysis of tissue extracts by 31P NMR and ion chromatography indicated that the most prominent components of the phosphomonoesters were glucose 6-phosphate, alpha-glycerol phosphate and AMP. In conclusion, of the various phosphorus metabolites that can be measured by 31P NMR, only one group, the phosphomonoesters, was predictive of functional recovery.


Nitric oxide is an important determinant of coronary flow in the isolated blood perfused rat heart. Many vasoactive substances are involved in the regulation of vasomotor tone and some of them, like nitric oxide (NO), are derived from the endothelium. Nitric oxide is able to relax preconstricted coronary resistance vessels almost completely. However, it is not clear what the contribution of NO is to vasomotor tone in the intact blood perfused heart. The aim of the present study was to evaluate the contribution of NO to coronary pressure-flow relations. We used isovolumically beating, donor supported, blood perfused isolated rat hearts. We measured pressure-flow relations under control conditions, after blocking endothelial NO production with NG-nitro-L-Arginine (LNNA) and after administration of L-Arginine (L-Arg) in order to overrule the blocking effect. Administration of LNNA at a perfusion pressure of 105 mm Hg resulted, after about 40 min, in a significant (Wilcoxon's signed-rank test, (n = 8) p < 0.05) reduction of coronary flow to 47 +/- 5% (mean +/- SEM) of control and a reduction of developed isovolumic left-ventricular pressure to 62 +/- 4% of control. L-Arg returned flow to 60 +/- 7% of control which is a significant increase with respect to LNNA (p < 0.05). L-Arg did not increase the left-ventricular pressure. The entire perfusion pressure-flow relation (pressure range 65-125 mm Hg) was significantly shifted downwards after LNNA with respect to control. Pressure-flow relations after L-Arg were in between those during control and after block of NO production. L-Arg alone was found to have no effect on flow and left-ventricular pressure (n = 2) and both LNNA and L-Arg were found to have no effect on contractility of isolated trabeculae (n = 6), thus, coronary blood flow reduction after LNNA administration is mainly the result of inhibition of endothelial NO production. At a perfusion pressure of 105 mm Hg reactive hyperemia is still present after LNNA and subsequent L-Arg administration, indicating that endothelial NO is not the only factor involved in flow regulation. We conclude that endothelium-derived NO is involved in the control of coronary flow in the blood perfused rat heart.


Preconditioning of heart by repeated stunning: adaptive modification of myocardial lipid membrane. Recent studies indicate that preconditioning of the heart by repeated stunning protects it from subsequent ischemic injury. Such myocardial preservation is likely to be due to adaptive modification of lipid composition and dynamic structure of cellular membrane. To test this hypothesis, swine heart was subjected to four episodes of 5 min stunning by LAD occlusion, followed by 10 min of reperfusion after each stunning. The heart was then made regionally ischemic for 60 min by LAD occlusion, followed by 6 h reperfusion. A control heart was perfused for 60 min, followed by 60 min ischemia and 6 h reperfusion. Free fatty acids (FFA) accumulated in the control heart during ischemia, as expected, which was further enhanced by reperfusion. The FFA level was also enhanced during ischemia in stunned myocardium. However, this FFA level was almost restored during reperfusion. The levels of unsaturated fatty acids (UFA) such as oleate, linoleate, and arachidonate followed a similar pattern. Membrane fluidity, monitored by fluorescence polarization, was decreased during ischemia and reperfusion in the unstunned heart, with the corresponding increase in microviscosity. The increased microviscosity was significantly reduced by stunning. Since FFA are presumably generated from membrane phospholipids, these results suggest that stunning may cause the incorporation of a greater proportion of PUFA in membrane phospholipids, leading to preservation of membrane phospholipids and maintaining the membrane fluidity, which may be at least partially responsible for the attenuation of ischemic reperfusion injury.


Sexual behavior, smoking, and HIV-1 infection in Haitian Women. To determine whether a previously observed association between human immunodeficiency virus type 1 (HIV-1) infection and smoking in Haitian women could be explained by confounding high-risk behaviors. A nested case-control study at a primary care health clinic in Cité Soleil, Haiti. Women who reported having smoked (n = 89) and randomly selected nonsmokers (n = 329) who had participated in a survey 1 to 12 months earlier evaluating risk factors for HIV-1 infection. Lifetime sexual practices, smoking, health beliefs and practices, and other factors potentially confounding the relationship between smoking and HIV-1 infection. Compared with nonsmokers, smokers reported higher rates of high-risk behaviors, including more lifetime sex partners (P less than .001), being less likely to be married (P less than .01), and being more likely to have visited folk healers (P less than .01). No intravenous drug use was reported, and no significant differences were noted between smokers and nonsmokers in numbers of past surgical or dental procedures or injections. Adjustment for all factors associated with HIV-1 infection and smoking in regression analyses revealed an independent association between smoking and HIV-1 infection (odds ratio [OR], 3.4; 95% confidence interval [Cl], 1.6 to 7.5). Other factors associated with HIV-1 infection included having more than two life-time sex partners (OR, 3.4; 95% Cl, 1.7 to 6.8) and lower socioeconomic status as reflected by a dirt floor in the home (OR, 8.6; 95% Cl, 3.3 to 22.0). Smoking is a marker for high-risk sexual behavior and is associated with an increased risk of HIV-1 infection in this population. The persistent association between smoking and HIV-1 infection after adjustment for all known risk factors suggests the possibility of a biologic effect of smoking that warrants further evaluation in other populations.


Effect of partial obstruction of the rabbit urinary bladder on malate dehydrogenase and citrate synthase activity. Previous studies have demonstrated that partial outlet obstruction in rabbits induced a significant decrease in oxidative metabolism in urinary bladder smooth muscle. The current experiments were designed to determine whether the decreased oxidative metabolism of obstructed bladder tissue is associated with alterations in the activities of specific mitochondrial enzymes. The activities of two important enzymes in the tricarboxylic acid cycle, malate dehydrogenase and citrate synthase, were measured in samples of bladder body and base from normal bladders and in bladders from rabbits in which partial outlet obstruction had been produced seven days prior to the experiments. The results can be summarized as follows: malate dehydrogenase activity was similar in bladder body and base isolated from control rabbits; and decreased by approximately 40% in both segments of the bladder isolated from obstructed rabbits. In contrast to malate dehydrogenase, citrate synthase activity was significantly higher in the bladder body than in the base of normal rabbits. Outlet obstruction caused about a 50% decrease in activity of this enzyme in the bladder body, but had no significant effect on citrate synthase activity of the bladder base. These findings demonstrate that the deficiency in bladder function following partial outlet obstruction is associated with a marked decrease in the activities of two essential enzymes in oxidative metabolism: malate dehydrogenase and citrate synthase. This decrease in enzyme activity is consistent with the previously observed decrease in oxidative metabolism and would be expected to lead to an inability of the tissue to supply sufficient metabolic energy for proper contractile function.


Antiproliferative effects of a c-myc antisense oligonucleotide on human arterial smooth muscle cells. The effects of a c-myc antisense phosphorothioate DNA oligonucleotide were assessed on the proliferation rate of human arterial smooth muscle cells (HSMCs). Compared to a control oligonucleotide the antisense oligonucleotide suppressed the proliferation of HSMCs in a concentration-dependent manner without a major cytotoxic effect. Outgrowth of HSMCs from media explants was significantly inhibited as well. Induction of c-myc expression by serum stimulation of cells was blunted by the antisense oligonucleotide, as shown by immunoblotting. These results demonstrate that c-myc expression is an essential factor for proliferation of HSMCs after growth stimulation, and they show the potential of antisense technology for modulating gene expression of HSMCs in vitro.


The reciprocal protective effects of magnesium and calcium in hyperkalemic cardioplegic solutions on ischemic myocardium. The cardioprotective effects of magnesium and calcium in hyperkalemic cardioplegic solutions were investigated in isolated rat hearts. Isolated rat hearts were arrested for 30 min at 37 degrees C in cardioplegic solutions containing magnesium and calcium in varying concentrations. In St. Thomas' Hospital cardioplegic solution, the magnesium and calcium concentrations were varied as follows: Mg 0; Ca 0.1, 0.3, 0.6, 1.2 mmol/l (mM)./ Mg 4; Ca 0.1, 0.3, 0.6, 1.2 mM./ Mg 8; Ca 0.1, 0.3, 0.6, 0.9, 1.2 mM./ Mg 12; Ca 0.1, 0.6, 0.9, 1.2 mM./ Mg 16; Ca 0.1, 0.6, 1.2, 1.5 mM. For each magnesium concentration, the percentage recovery of aortic flow generated dose-response curves depending on calcium concentration. The maximum percentage recovery of aortic flow was 76.0% +/- 2.7% (mean +/- SEM) in the Mg0-Ca0.1 mM group, 77.1% +/- 2.0% in the Mg4-Ca0.3 mM group, 78.5% +/- 2.3% in the Mg8-Ca0.6 mM group, 79.8% +/- 2.4% in the Mg12-Ca0.9 mM group and 80.0% +/- 3.4% in the Mg16-Ca1.2 mM (ST solution) group. Significant difference in the recovery of aortic flow has not been observed among these groups, and furthermore, significant differences in the recovery of other parameters of cardiac function and Ck leakage have not been observed among these groups. These results suggest that the cardioprotective effects depend on the relative combination of magnesium and calcium concentrations, and that it is important to maintain an appropriate balance of magnesium and calcium in hyperkalemic cardioplegic solutions.


The electrophysiology of cardiac allograft rejection: independent effects of rejection and perioperative ischemia on the sinus node recovery phenomenon after cardiac transplantation. We characterized the effect of cardiac allograft rejection on the sinus node (SN) recovery response from overdrive suppression. A total of 54 corresponding data sets (SN recovery time [SNRT]/endomyocardial biopsy [EMB]) was available in 24 transplant recipients with normal SNRT. Data were pooled in the rejection vs the no-rejection group (n = 16 vs n = 38, respectively). During cardiac rejection (defined as a 7-day period starting 3 days prior to and lasting until 3 days after the EMB) the SNRT curves were moderately, but significantly shifted towards higher values (F = 13.4, p = .0003). All changes occurred within accepted normal limits for the SNRT. Multivariate analysis indicated independent effects of donor heart ischemic time (p = .0005) on SNRT in addition to that of rejection. After accounting for that influence of ischemic time respective F values regarding the influence of rejection on the SNRT excursions were 10.8 (ischemic time < 100 min, p = .0014) and 4.36 (ischemic time > or = 100 min, p = .039). This study shows that cardiac allograft rejection significantly delays the SN recovery response from overdrive suppression. These changes, however, are subtle and, hence, are an unlikely explanation for the often grossly abnormal postoperative SN function.


A microassay for proteases using succinylcasein as a substrate. A photometric assay for proteases has been developed. A chemically modified casein whose amino groups were succinylated was used as a substrate. After incubation with trypsin, chymotrypsin, thermolysin, and subtilisin, the extent of hydrolysis of the substrate was determined with trinitrobenzene sulfonate (TNBS). The whole procedure of the assay was performed in the microtiter plate wells and the increase in the absorbance resulting from the reaction between TNBS and newly formed amino groups in the substrate was able to be determined with a high sensitivity by a microtiter plate reader, enabling the simultaneous measurement of a number of samples. Application of this method to the measurement of proteolytic activity contained in the protein extract of Tapes philippinarum is demonstrated.


Differential effects of quinidine and flecainide on plateau duration of human atrial action potential. Quinidine and flecainide, two class-I antiarrhythmics increase action potential duration (APD) at 90% repolarization and cellular refractory period in human atrial fibers without significant change in resting potential. On the other hand, quinidine decreases APD at 50%, whereas flecainide slightly increases, which suggests different effects on Ca2+ current. Using isolation cell procedure and whole cell recording, we found that 10 microM quinidine (34.77 +/- 6.5%, n = 5) and 0.5 microM flecainide (50.46 +/- 6.2%, n = 4) decrease calcium current in human atrium. It is concluded that, at therapeutical concentrations, quinidine and flecainide modify the action potential plateau phase in a different manner, which is not only related to the calcium current decrease.


A genetic abnormality of cardiac myocytes from the blind mutant (RC) chick heart: abnormalities of cardiac structure and choline transport. A new genetic cardiomyopathy was identified in a blind mutant avian strain. Cardiac myocytes were cultured from 7-day-old chick embryos from Rhode Island Red chickens and from another strain of this species that has been identified to have several abnormalities, the most striking of which is blindness. Cardiac myocytes were maintained in tissue culture. Morphologically, in culture, the cardiac myocyte from the blind mutant strain assumed a spherical shape and showed abnormalities of sarcolemmal membrane compared to control myocytes from heterozygous animals. Choline uptake and metabolism were examined, using [methyl 3H] choline, because it is a sarcolemmal transport process and choline is metabolised to phosphatidylcholine, an important phospholipid for cellular structure and function. Choline uptake through the active transport process was markedly and significantly reduced in the mutant cells compared to control cells, while choline metabolism to phosphatidylcholine was not significantly altered. These results demonstrate a new abnormality of cardiac myocytes, a cardiomyopathy that can be studied in cell culture and one with abnormalities of cellular choline transport.


Combined immunohistochemical and immunofluorescence method to determine the phenotype of proliferating cell populations. To determine the phenotype of proliferating cell populations. The double immunostaining technique combines the autofluorescent properties of alkaline phosphatase substrate naphthol/Fast Red with immunofluorescence using fluorescein. Fresh human tonsil and fresh atherosclerotic aortic aneurysm wall tissue were studied using a panel of monoclonal antibodies including Ki-67, CD4, CD8, CD19, CD22, HLA-DR alpha, CD68 and CD31. This double immunostaining method permitted simultaneous colocalisation of different markers on the same cell and could be used to identify HLA-DR positive cells as well as proliferation associated Ki-67 positive cells in human tonsil tissue and in chronic periaortitis associated with advanced atherosclerosis. This technique is simple and the results may be viewed using a single fluorescence filter. The Fast Red reaction product is stable and does not fade under storage. The staining works particularly well with markers for nuclear antigens in combination with markers for cytoplasmic or surface antigens.


Various wedge isodose angles for treatment planning. Various wedge isodose angles or simply wedge angles smaller than the nominal wedge angle were created by combining the isodose distributions generated from a single physical wedge with the isodose distributions of the open field for the 8-MV photon beam. The particular wedge angle generated depends on the weights imposed on these isodose distributions. The relationship between these weights and the wedge angle were examined and found to be nonlinear. The difference between the wedge angles defined at 10 cm depth and those defined using the 50% isodose curve is less 6 degrees. The present data was fitted using two proposed empirical equations.


Wedge filter effects on dosimetric parameters of a linear accelerator. The open-field and wedged-field output factors as a function of field size for two linear accelerators were measured. Wedge factors were determined by taking the ratio of the outputs with and without the wedge filter. For one linear accelerator, the difference in the output factors between the wedged field and open field can be as large as 5%. The wedge factor for this linear accelerator also varies with the field size. On the other hand, the other linear accelerator shows smaller variation of output factors between wedge field and open field. The variation of wedge factor is less than 1% for a 60 degree wedge. In addition to modifying the isodose distributions, the wedge filter also changes the percent depth dose curves, the output factor, and the wedge factor. The degree of wedge effects on these dosimetric parameters is different for different linear accelerator.


A dosimetric study comparing three-, four-, and six- field plans for treatment of carcinoma of the prostate. We present a three-dimensional dosimetric analysis of 3-, 4-, and 6-field plans using 24 MV photon beams for treatment of carcinoma of the prostate. We compare isodose distributions on a transverse plane through the center of the target as well as differential and integral dose volume histograms for the target and critical structures, respectively. An extensive study on a representative case led to the development of a technique where two complementary 3-field daily plans deliver the same daily target dose as the standard 4-field box while affording sparing of the bladder and rectum similar to that achieved with a 6-field plan. This technique was shown to yield the same results on a sample of four additional patients representing a range of target and patient sizes. We conclude that the combined two-day, 3-field method for treatment of the prostate may be a better choice than the standard 4-field box or the 6-field daily plan for dose escalation studies.


Three-dimensional dosimetric comparison of radiation therapy treatment planning of the pancreas. Radiation therapy for carcinoma of the pancreas requires high doses for local control. Three-dimensional dose distributions are calculated for four patients with pancreatic cancer using two conventional (4-field box and 3-field techniques) and two noncoplanar (4 and 6 oblique fields) treatment field arrangements. Uniform dose distributions are obtained for all beam arrangements. The 4-field oblique beams show a potential advantage for lower dose to the kidneys when compared with 4-field box technique, and to the small bowel, when compared with 3-field beams. The data suggest that the noncoplanar beams may be useful alternative techniques for treating this disease with certain case presentations and should be considered during the treatment planning process.


Comparison of absorbed doses in water and solid water for electron beams. Solid water, as a substitute for water, has become commercially available for dosimetry measurements. A study was undertaken to compare the dose in water and solid water respectively for 6, 9, 12, 16 and 20 MeV electron beams. Measurements using ion chamber show that the dose in water is higher than the dose in solid water by 1% for 6, 9, and 12 MeV electrons. For 16 and 20 MeV electrons, the dose in water and solid water are the same within the uncertainty of our measurements.


Dosimetry of shaped electron fields using a radial integration method. The feasibility of an analytical approach to calculate monitor units for shaped electron fields is investigated. A radial integration method is used to calculate the dose at prescription depth from an average output factor and an average depth dose. This concept, as implemented in a commercial planning system, has been tested on various arbitrary fields, and 66 shaped electron fields clinically used for head and neck, chestwall, internal mammary, breast boost, and skin lesions. The measured and prescribed doses agreed within 3.5% or better for 71% of all clinical fields tested; for 91% the agreement was 5.5% or better. The greatest discrepancy (-7.2%) was found for a narrow, long internal mammary field. All treatments were administered on a linear accelerator with electron energies between 6 and 20 MEV.


A simple method for indirectly verifying noncoplanar treatment fields in radiotherapy. The use of a vertex field can be a practical technique for treatment of pituitary and brain tumors. This approach may be underutilized, primarily because of the difficulty in documenting noncoplanar treatment fields. This paper describes a method for documenting vertex treatment fields by inference using portal films. This method is relatively simple to implement, yet yields accurate results.


Advantages of using high activity 125I seeds in temporary interstitial breast implants. There has been considerable interest in the use of high activity 125I sources as a substitute for 192Ir seeds for removable implants of the breast and prostate. 125I seeds with an initial activity of approximately 5 mCi per seed, loaded in special afterloading nylon catheters, are used to improve dose distribution in the tumor volume and minimize dose to the adjacent critical organs and normal tissues. Seed spacing in strands is adjusted to maintain a dose rate of 40-60 cGy per hour at a distance of 5 mm from the plane of the implant. Implants custom loaded with 125I sources achieve superior isodose distribution compared to implants loaded with standard 192Ir seed strands. High activity 125I seeds also offer the advantage of reduced exposure to radiation oncology staff, nurses, and visitors leading to better patient care. Due to reduced exposure to personnel, the accuracy of the actual implant geometry can be verified by taking a localization film with actual 125I sources placed in the tumor bed.


A simple eye-lens shielding technique compatible with independent jaws of a linear accelerator. A technique, compatible with accelerators having independent collimators, is presented, which greatly simplifies alignment of pencil eye-lens shields. The patient is placed so that the eye to be shielded is on the central axis of the beam and the accelerator's collimators are adjusted independently to define the field. A divergent lead shield 14 cm long is suspended from a wire that hangs along the central axis from a blocking tray in its usual location. The suspension length is variable and adjusted on the first day so that the shield is about 1 cm above the patient's eye. On subsequent treatment days, the wire is simply put in place on the tray with no further adjustment required. This technique reduces setup time and makes the shield positioning highly reproducible.


Photon beam attenuation for a patient support assembly during arc therapy for a medical linear accelerator. Arc therapy is one of the treatment techniques for small, centrally located deep-seated tumors. However, care must be taken to remove any components that would interfere with the beam as the gantry rotates around the patient. One such component that may interfere with the beam is the patient support assembly (PSA) or treatment table. Beam attenuation factors due to the presence of the couch side-rails and the centerspine bar of the PSA are presented for both 6 MV and 18 MV photon beams of a Clinac 1800 during a 360 degrees rotation. Dose perturbations arising from these obstructions are displayed using an Alderson Rando phantom. A method is described to calculate the start and stop angles for the largest unobstructed arc for a given field size (FS), centerspine bar to the isocenter height (H), centerspine bar width (W), and SAD. As an illustration, for an SAD of 100 cm and a W of 4 cm, the start and stop angles for arc therapy with a FS of 10 x 10 cm2 and an H of 12.7 cm would be 29 degrees and 331 degrees.


Mutational specificities of N-nitrosamines in a host-mediated assay: comparison with direct-acting N-nitroso compounds in vitro and an approach to deducing the nature of ultimate mutagens in vivo. The mutational activities and specificities of several N-nitrosamines in Salmonella recovered from mouse liver in the host-mediated assay (HMA) were compared with the specificities of related direct-acting N-nitroso compounds in vitro. The specificities of the direct-acting methyl, ethyl, propyl, and 2-hydroxypropyl compounds were all different and presumably are attributable to the DNA adducts resulting from the corresponding alkyldiazonium or carbonium ions. Introduction of a 2-hydroxyl group greatly influenced the mutational specificity. The 2-oxopropyl compound showed the same specificity as the methyl compound. This result is consistent with one of the known breakdown pathways of the oxopropyl diazonium ion (or related reactive species), which leads to a methyl diazonium ion. The N-nitrosodialkylnitrosamines N-nitrosodimethylamine (NDMA), N-nitrosodiethylamine (NDEA), and N-nitrosodipropylamine (NDPA), which all require metabolic activation, showed specificities in the HMA similar to those of their direct-acting counterparts. The cyclic nitrosamine N-nitrosopyrrolidine was weakly active in the HMA, although its direct-acting derivative was a potent mutagen in vitro. The results for NDMA and NDEA were consistent with most previous studies of the metabolism of these compounds in vivo. However, NDPA can yield methylating, and probably hydroxypropylating, species in addition to propyldiazonium ion. As the specificity of NDPA was similar to that of a propylating agent, NDPA appears to lead to genotoxic products in the mouse liver mainly by direct alpha-hydroxylation. The initial results described here indicate that mutational specificity in the HMA can be used to deduce metabolic pathways leading to genotoxic products when the appropriate proximate mutagens are available as standards. Furthermore, we observed a reasonable correlation between potency in the HMA and hepatocarcinogenesis.


Suppression of radiation-induced neoplastic transformation by overexpression of mitochondrial superoxide dismutase. Manganese superoxide dismutase (MnSOD) scavenges toxic superoxide radicals produced in the mitochondria. Transfection of the human MnSOD gene into mouse C3H 10T1/2 cells resulted in production of active MnSOD, which was properly transported into mitochondria. Overexpression of MnSOD protected cells from radiation-, but not chemically-induced neoplastic transformation. This finding demonstrates that oxidative stress that occurs in the mitochondria plays an important role in the development of neoplastic transformation.


Partial suppression of tumorigenicity in a human lung cancer cell line transfected with Krev-1. A human non-small-cell lung carcinoma cell line, Calu-6 (from an anaplastic carcinoma), was transfected with the Ki-ras-related anti-oncogene Krev-1. Several transfectant lines were obtained that showed a reduced tumorigenicity in nude mice with respect to the parental and control transfected cell lines. This decrease was approximately 50% in tumor incidence at 4 wk after subcutaneous inoculation of the transfected cells. In addition, the volume of the Calu-6 revertant-derived tumors was three to 10 times smaller than that of the equivalent tumors produced by inoculation of the control cell line transfected with the neomycin-resistance gene. Krev-1--transfected cells that exhibited reduced tumorigenicity expressed Krev-1 mRNA and had variable numbers of copies of the Krev-1 gene. Moreover, Krev-1--transfected cells exhibited a more differentiated squamous epithelial morphology than the parental and control cell lines did. Moderately elevated levels of protein kinase C activity were detected in some revertant clones. Such activity correlated with the level of expression of Krev-1 mRNA in most cases. In summary, Krev-1 induced important morphological and biological changes in transfected Calu-6 cells that we interpreted as partial reversion of the malignant phenotype.


ras oncogene activation in mammary carcinomas induced by N-methyl-N-nitrosourea in Copenhagen rats. The occurrence of Ha-ras and Ki-ras oncogenes was investigated in mammary tumors produced by treating genetically resistant Copenhagen (Cop) rats with N-methyl-N-nitrosourea. G35-->A codon 12 mutations in both Ha-ras and Ki-ras genes were analyzed by a polymerase chain reaction/liquid hybridization and gel retardation assay. More than half of the adenocarcinomas analyzed contained an activated Ha-ras gene. This was also the predominant mutation in similar tumors from susceptible Buf/N rats, suggesting a common mechanism of initiation. In contrast, only two of 15 mammary adenosquamous carcinomas from the Cop rats contained an activated Ha-ras gene, suggesting a different initiation mechanism for most of these tumors. Ki-ras activation was found in none of five and one of five adenocarcinomas from Buf/N and Cop rats, respectively, and in none of 13 adenosquamous carcinomas from Cop rats. These results suggest that Ki-ras activation does not play a major role in the initiation of the mammary tumors.


Growth and transformation suppressor genes for BHK Syrian hamster cells on human chromosomes 1 and 11. To map putative tumor suppressor genes for the near-diploid baby hamster kidney fibrosarcoma cell line BHK, we transferred five different normal human chromosomes (1, 3, 7, 11, and 12) into these tumor cells by microcell-mediated chromosome transfer. Transfer of human chromosome 1 into BHK cells resulted in suppression of cell growth both on plastic and in soft agar, indicating that chromosome 1 has a generalized effect on cell growth and thereby suppresses anchorage-independent growth. Selection against cells with an intact chromosome 1 was observed. In contrast, the introduction of chromosome 11 into BHK cells resulted in suppression of anchorage independence but not growth on plastic. Most chromosome-11 growth-suppressed BHK hybrids retained intact copies of human chromosome 11. Tumorigenic derivatives of chromosome 11 hybrids had lost this chromosome. Transfer of human chromosome 3, 7, or 12 into BHK cells did not correlate with growth suppression of BHK cells on plastic or in soft agar. Thus, we conclude that genes that suppress BHK-cell growth in general or in agar reside on human chromosomes 1 and 11, respectively.


[Dimensions of the human aortic arch and thoracic aorta studied by nuclear magnetic resonance]. The size of the aortic arch and thoracic aorta was measured in 50 healthy volunteers studied by NMR. In each case the following parameters were measured and statistically elaborated: diameter (in mm) of ascending aorta (between onset and arch beginning), of aortic arch (intermediate zone), of isthmus, of descending aorta (between isthmus and diaphragm), of diaphragmatic isthmus; a mean diameter was also calculated, and for every diameter mean and standard deviation were obtained. Moreover, the length (in mm) of superior and inferior contours of the aortic arch, as the ratio of these two values were obtained. At last, total length of considered aortic tract, area of visualized surface and theoretic volume were obtained. For each parameter a diagram was then plotted, with age in ordinates and values of above mentioned parameters in abscissae. Statistical evaluation allows to affirm that the size of the aorta grows with ageing. On the contrary, the ratio between superior and inferior contour of the aortic arch tends to remain unvaried; this fact seems to disagree with the current opinion that the superior wall of the arch grows with age more than the inferior one.


[Structural organization of the menisci of the human knee: scanning electron microscopy study]. The structure of the meniscus of the knee joint was studied in 10 human subjects after meniscectomy for acute trauma. The specimens were studied by scanning electron microscopy after cryofracture. Collagen fibres are arranged in two layers: a thin superficial zone with radially oriented fibres and a deep zone with circumferentially oriented ones. Deep fibres are surrounded by oblique fascicules. The cells are rare and oval-shaped. The observations enhance the hypothesis that the meniscus of the knee joint is not a real fibrocartilage.


Mechanical properties of the coronary vasculature: indirect evaluation. Information on the mechanical properties of the coronary vascular bed can be obtained indirectly by modelling the vascular system. This indirect approach, unlike 'in vitro' measurements, allows to take into account the vasomotor conditions of the circulatory district as well as the effect of the surrounding embedding tissue on the vascular performance. An experimental manoeuvre of sudden occlusion and subsequent release of the thoracic descendent aorta on 5 anaesthetized dogs with open pericardium induces a step-like variation in the coronary perfusion pressure and the occurrence of oscillations in the mean coronary flow. Such a behaviour can be described using a second-order model ('windkessel'+inductance, which takes into account blood inertia in the large vessels). The value of the coefficients entering the equations have been obtained with a 'best-fit' procedure (minimum of the chi-squared variable) on the haemodynamical data. Coefficient variations are in agreement with the direct estimation of the myocardial compliance and volume, measured by Ultrasound Echocardiographic imaging (4-chamber projection mode).


[High-conductance K+ channels in guinea pig gallbladder epithelium]. Since secretion of electrolytes may be regulated by membrane potential difference, ion channels were studied using patchclamp technique. We have identified, in cell-attached configuration, inward-rectifying channels: the zero-current potential corresponded to the K+ equilibrium potential calculated from intracellular K+ activity. Using inside-out configuration and symmetric 145 mM KCl salines, i/V curve was linear, channel conductance was about 170 pS and the reversal potential 0 mV. The channels were selective for K+ over Na+, N-methylglucamine and anions and were activated by membrane depolarization.


Fertilization and embryonic development in vitro in cattle. Morphological and cytogenetic observations. The presented ichonography provides a documentation of the changes occurring during maturation, fertilization of oocytes and embryonic development in vitro; it points out the correlations between morphological features and the stage of nuclear chromatin and gives the possibility to observe some abnormalities that frequently occur during these processes.


The reliability of eye movement quantitative evaluation. I. Saccadic eye movements. We adopted the estimate of the intraclass coefficient of reliability, R, to evaluate the reliability of saccadic eye movements quantitative analysis. At a one-week interval we recorded refixative saccadic eye movements twice from fifteen healthy subjects by means of the binocular electrooculographic technique. R was computed for the constants and the slopes of the amplitude/duration and the amplitude/peak velocity relationships, for the mean precision values and for the mean latency values adjusted for subject's age. Our data demonstrated that the reliability of saccade parameters is fairly good for the amplitude/peak velocity relationship, good for the precision and very good for the amplitude/duration relationship. Finally, we believe that the normal variability values we obtained can be usefully employed in neurophysiological longitudinal studies not only in normal subjects, but also in pathological condition provided the more reliable parameters and the more adequate strategies to compute normal variability values are chosen.


The reliability of eye movement quantitative evaluation. II. Smooth pursuit eye movements. We adopted the estimate of the intraclass coefficient of reliability, R, to evaluate the reliability of smooth pursuit eye movement quantitative analysis. At a one-week interval, we recorded twice smooth pursuit eye movements from fifteen healthy subjects by means of the binocular electrooculographic technique. R was computed for the constant and the slope of the target velocity/gain relationships. R values were rated good for the slope and excellent for the constant. Finally, we computed for each parameter the maximum variability value according to two differing methods; on the basis of the within-subjects mean square values, we defined the normal range of biological test-retest variability for the two parameters.


The effect of daily hassles on salivary IgA: experimental evidence. The study aimed at evidencing the suppressive effect of daily hassles on salivary immunoglobulin A (S-IgA); in fact, previous research did not detect a clear relationship between the two variables. Twenty-four subjects were tested as to the hassles that had occurred in their recent life and to S-IgA; the tests were repeated three times, with an interval of four weeks between each. The results show that the variation of S-IgA values between times 2 and 3 is associated with the variation of hassles between times 1 and 2 (r = -.46, p < .01).


Autogenic training and signs of distress: an experimental study. The research investigated the autogenic training (AT) effects, analyzing the emotional and somatic distress symptoms of patients during their learning of the AT standard exercises (N = 79). An improvement was observed even before the beginning of the course, thus demonstrating the Balint effect of the drug-doctor phenomenon. The control subjects, entered in a waiting list, showed no further improvements from this phase of the study; the experimental ones, who had begun their AT exercises, continued to show significant improvement.


Assessment of functional ability in Alzheimer disease: a review and a preliminary report on the Cleveland Scale for Activities of Daily Living. Assessment of activities of daily living (ADL) in Alzheimer disease (AD) is critical in establishing the diagnosis, monitoring disease progression, evaluating the efficacy of treatment interventions, and determining the need for health and social services. The proper method to measure ADL depends on the purposes to which the scale is to be put. Existing ADL scales differ as to the type of behaviors assessed, the nature of the observations made, and the manner in which the observations are quantified. These scales were not specifically designed to evaluate changes in the nature and extent of the broad spectrum of functional difficulties seen in individuals with AD. We describe the Cleveland Scale for Activities of Daily Living (CSADL), an informant-based instrument designed to expand upon the capacity of existing physical and instrumental ADL scales by assessing both premorbid and current component acts (e.g., initiation versus implementation) of daily living functions.


Alzheimer Disease Assessment Scale: a subtest analysis. The Alzheimer Disease Assessment Scale (ADAS) was administered to 61 Alzheimer patients, 52 elderly controls, and 80 controls between age 7 and 54 years. The Alzheimer group was categorized into different severity levels of dementia based on MMSE scores: very mild (> or = 24), mild (> or = 20), moderate (10-19), and severe (0-9). All 11 ADAS Cognitive subtest scores for the mild, moderate, and severe dementia groups were statistically worse than the elderly control group. This was also the case for the very mild group, except for Naming, Commands, Constructional Praxis, and Ideational Praxis. In terms of magnitude of effect, memory and spontaneous language items were the earliest indicators on the ADAS, while praxis, commands, and naming items were only sensitive later in the course of the disorder. The best single indicators of progression throughout the severity continuum of dementia (i.e., from normal to severe) were the Orientation subtest, the ADAS Cognitive score, and the ADAS Total score. The ADAS Noncognitive subtests generally did not show the progression with increasing dementia that was evident on the ADAS Cognitive subtest. Differences in educational level had no statistically significant effects on any of the ADAS subtest scores, and age differences were few and small in magnitude. The differential rate of decline of the various ADAS subtests appears to reflect both the changing pattern of cognitive impairments as a function of severity of DAT and also to some extent the psychometric limitations of some of the subtests.


Colony-stimulating factors: a new step in clinical practice. Part I. Haematopoietic colony stimulating factors (CSFs) stimulate the proliferation, differentiation and functional activity of myelocytes. Since approval of the use of these factors in patients with chemotherapy-induced neutropenia, patients undergoing autologous bone marrow transplantation (BMT) and patients with delayed engraftment after BMT, several other potential clinical uses have been described. Current and potential future clinical applications of CSFs will be reviewed.