Ergonomic research study on aircraft luggage handling. In this study, the lifting and carrying of freight and luggage at a large German airport is classified by typical conditions of performance. These manual load handling tasks take place in height-restricted workplaces. Different methods for an ergonomic evaluation of these tasks are used. The results of the epidemiological, physiological, and biomechanical studies indicated a strain bottleneck during loading tasks in aircraft holds. Therefore, an additional laboratory study was done, in order to analyse the influence of work weight and weight of work object on heart rate, subjective RPE, and number of work objects handled. As a result of the study, the airport has developed a special lifting and carrying training for the aircraft handlers which aims to reduce strain on the postural and locomotor apparatus by an appropriate technique of work. The training programme consists of theoretical instruction by a company doctor and practical exercises under the supervision of a specially-trained sports teacher.


Suppressor analysis suggests a multistep, cyclic mechanism for protein secretion in Escherichia coli. The sec/prl gene products catalyze the translocation of precursor proteins from the cytoplasm of Escherichia coli. Recessive, conditionally lethal mutant alleles of these genes (sec mutations) cause a generalized defect in protein secretion; dominant suppressor mutant alleles (prl mutations) restore export of precursor proteins with altered signal sequences. In prl strains, a precursor protein with a defective signal sequence can be selectively targeted to the suppressor gene product. When a precursor LacZ hybrid protein is used, the targeted prl protein is inactivated by the large, toxic hybrid molecule, a result termed suppressor-directed inactivation (SDI). Using SDI, two different secretion-related complexes can be generated: a pretranslocation complex that contains a hybrid protein with an unprocessed signal sequence, and a translocation complex in which the hybrid protein is jammed in transmembrane orientation with the signal sequence cleaved. Additional Sec proteins that are contained within, and thus sequestered by, each of these complexes can be identified when their functional levels are lowered using the conditional lethal sec mutations. Results of this genetic analysis suggest a multistep pathway for protein secretion in which the translocation machinery assembles on demand.


Conversion of human interleukin-4 into a high affinity antagonist by a single amino acid replacement. Interleukin-4 (IL-4) represents a prototypic lymphokine (for a recent review see Paul, 1991). It promotes differentiation of B-cells and the proliferation of T- and B-cell, and other cell types of the lymphoid system. An antagonist of human IL-4 was discovered during the studies presented here after Tyr124 of the recombinant protein had been substituted by an aspartic acid residue. This IL-4 variant, Y124D, bound with high affinity to the IL-4 receptor (KD = 310 pM), but retained no detectable proliferative activity for T-cells and inhibited IL-4-dependent T-cell proliferation competitively (K(i) = 620 pM). The loss of efficacy in variant Y124D was estimated to be greater than 100-fold on the basis of a weak partial agonist activity for the very sensitive induction of CD23 positive B-cells. The substitution of Tyr124 by either phenylalanine, histidine, asparagine, lysine or glycine resulted in partial agonist variants with unaltered receptor binding affinity and relatively small deficiencies in efficacy. These results demonstrate that high affinity binding and signal generation can be uncoupled efficiently in a ligand of a receptor belonging to the recently identified hematopoietin receptor family. In addition we show for the first time, that a powerful antagonist acting on the IL-4 receptor system can be derived from the IL-4 protein.


The carboxy-terminal exon of the adenovirus E1A protein is required for E4F-dependent transcription activation. The adenovirus-2 E1A 289R transcription activator protein contains a 49 amino acid sequence (designated CR3) that has been suggested to represent the minimal domain required for E1A-induced activation of viral early transcription. We show here that the non-conserved carboxy-terminal E1A exon contains two interchangeable elements that are required for efficient CR3-dependent transactivation of the adenovirus E4 promoter in HeLa cells. These two elements do not encode independent transactivation functions and have been designated auxiliary regions (ARs) 1 and 2. The effects of AR1 and AR2 are not additive, suggesting that they contribute a mechanistically analogous function in transcription. Previous studies have suggested that two cellular transcription factors, ATF-2 and E4F, can function together with E1A to induce transcription of the E4 promoter. The importance of respective factors for E4 transcription has not been resolved. We find that E1A activation of E4F, but not ATF-2 (or other ATF factors), is AR1- and AR2-dependent. This result suggests that E1A induction of the E4 promoter in HeLa cells is primarily mediated by E4F.


Inter-study variability in left ventricular mass measurement. Comparison between M-mode echography and MRI. In order to compare variability in M-mode echography and MRI in the assessment of left ventricular mass, 20 echogenic patients without evidence of coronary artery disease were investigated. Two MR and two M-echo examinations were performed within 4 days by different trained operators, each unaware of the other's results. M-mode echo was carried out according to Devereux's method, using the 'Penn-Cube' formula. MR protocol included multislice (8 to 12) true, short-axis spin-echo imaging (10 mm thick with a 1 to 3 mm gap) encompassing the entire left ventricle. Planimetry was manually traced with standardized window settings. Correlations between both echographic and both MR measurements showed r = 0.89, SEE = 22.7 g and r = 0.96, SEE = 11.2 g, respectively. Mean inter-study variability was 11 +/- 6.4% and 6.75 +/- 3.8% (P = 0.0021). The threshold value corresponding to the 95th percentile of the variability data was 21.5% for echography and 13.5% for MR. In conclusion, MR appeared to be a significantly more reproducible examination tool, when compared with M-mode echo, for the evaluation of left ventricular mass (variability, 63% higher with echo than with MR). The main practical consequence of this result lies in the reduced number of patients required to demonstrate a significant change in the LVM with MR as compared with echography.


Relative efficacy of angiotensin converting enzyme inhibitors on mortality of patients with congestive heart failure: implications of randomized trials and role of the aetiology (ischaemic or non-ischaemic) of heart failure. We examined the influence of angiotensin converting enzyme inhibitors (ACE inhibitors) on mortality in patients with heart failure of both ischaemic or non-ischaemic origin. Eleven, randomized, placebo-controlled trials of ACE inhibitors involving 1266 patients were selected. The follow-up period varied from 3 to 6 months. Four different ACE inhibitors were used in the 11 clinical trials. A total of 679 patients presented with an ischaemic heart failure and 587 with a non-ischaemic heart failure. Meta-analysis, performed for both subgroups, showed that mortality was significantly decreased in the ischaemic subgroup only (ischaemic group: odds ratio 0.45; 95% confidence interval 0.28 to 0.71; non-ischaemic subgroup: odds ratio 0.7; 95% confidence interval 0.4 to 1.5). Although the two odds ratio are not significantly different, further randomized, placebo-controlled trials with ACE inhibitors are required in order to determine more precisely the benefit/risk ratio in patients with non-ischaemic heart failure.


Distribution of two types of infiltrating helper T cells among rat renal allografts, spleen, peripheral blood, and regional lymph node cells. Cytokine production by graft-infiltrating cells (GIC) of helper T cell (Th) subtypes were investigated in rat kidney allograft compared to those infiltrating the spleen, those in peripheral blood, and in regional lymph nodes. The relative proportion of OX-22 positive cells (Th type 1) and the W3/25-positive cells (Th) of GIC was 49.4 +/- 2.1% on day 3 and 52.7 +/- 2.2% on day 6, whereas those in spleen cells was 80.3 +/- 4.1% (p less than 0.05) and 95.5 +/- 2.5% (p less than 0.01), respectively. GIC produced more B cell-stimulating factor 2 (BSF-2, 106.3 +/- 8.2 U/ml) when compared to spleen cells (15.9 +/- 4.6 U/ml; p less than 0.01), peripheral blood mononuclear cells (4.2 +/- 1.0 U/ml; p less than 0.01) or lymph node cells (24.9 +/- 4.0 U/ml; p less than 0.01) upon stimulation with donor BN lymphocytes in vitro. However, GIC produced less interleukin 2 (IL-2, less than 1.0 U/ml) and IL-3 (less than 1.0 U/ml) than produced by spleen cells (5.1 +/- 0.9 U/ml, p less than 0.05, and 10.5 +/- 2.3 U/ml, p less than 0.01, respectively) or lymph node cells (4.5 +/- 0.4 U/ml, p less than 0.01; 23.5 +/- 3.1 U/ml, p less than 0.01, respectively). These results demonstrate that the subtype of Th cells which produce BSF-2 migrates selectively to allografts.


Therapeutic lidocaine concentrations have no effect on blood platelet function and plasma catecholamine levels. The effects of therapeutic plasma concentrations of lidocaine on blood platelet function and plasma catecholamine levels were assessed in 9 healthy subjects. There were no significant effects on plasma levels of beta-thromboglobulin, collagen and adenosine diphosphate-stimulated platelet aggregation, thromboxane-B2-concentration in plasma after collagen and ADP stimulated platelet aggregation, or on plasma nor-adrenaline and adrenaline. No significant correlation could be demonstrated between any of the variables tested. Thus, it appeared that lidocaine had no effect on platelets that could be of benefit in acute myocardial infarction. It should be possible to use lidocaine, in combination with thrombolytic therapies without increasing the risk of bleeding complications.


[Age-related non-tumorous lesions in SPF C57BL/6 mice with special reference to amyloidosis]. Non-tumorous pathological changes in C57BL/6 CrSlc mice, which were reared under a barrier system and died spontaneously, were examined. At 3 months intervals 125 to 209 mice were purchased at 4 weeks of age and raised for the supply of aged animals. A large portion of the mice were used for various experiments between 3 and 30 months of age, while not a small number died spontaneously and were autopsied. The major non-neoplastic lesion was amyloidosis, with incidence of 55.5% and 74.4% for the autopsied female and male, respectively. The organs involved were the liver, kidneys, spleen, adrenal glands, ileum, heart and lungs. Skin ulceration and its scar, cerebral vascular calcification, glomerulosclerosis and sepsis in both sexes, distension of the seminal vesicles in males, fibroblast growth of the adrenal glands in females were commonly found. Incidence of spontaneous neoplastic lesions was 69.7% and 55.1% for the female and male, respectively.


Fine structure of atrial natriuretic peptide (ANP)-granules in the atrial cardiocytes in the hamster, guinea pig, rabbit, cat and dog. In the hamster, guinea pig, rabbit, dog and cat, the right and left atria and ventricles were examined by immunohistochemistry, and the right auricular cardiocytes were studied by transmission electron microscopy. Moreover, ANP-granules in the cardiocytes were analyzed by ultrastructural morphometry. Immunohistochemically, the most intensely ANP-reactive cardiocytes were localized in the right auricle, particularly more prominent in the hamster and guinea pig than in the rabbit, dog and cat. The immunoreaction in the dog and cat was weaker than that in the rabbit. ANP-immunoreactivity was not detected in the ventricular myocardium of any of all species examined, but was occasionally observed in the subendocardium of the ventricular septum. Ultrastructurally, ANP-granules were localized principally in the perinuclear region associated with the Golgi apparatus and scattered throughout the sarcoplasmic layers. The Golgi apparatus of the cardiocytes was better developed in the hamster and guinea pig than in the rabbit, dog and cat. It was poorly-developed in the dog and cat. By ultrastructural morphometry, the number of granules was greatest in the hamster followed by the guinea pig, rabbit and dog or cat, in this order. On the other hand, the diameter of granules was largest in the guinea pig and reduced via the hamster to the rabbit. The diameter was significantly smaller in the dog than in the rabbit. The diameter of granules of the cat was lay between the rabbit and dog.


[A trial designed to obtain a specific pathogen free Syrian hamster colony by administration of chemicals]. Conventional Syrian hamsters, contaminated with Giardia spp., Spironucleus muris, Trichomonas spp., Pasteurella pneumotropica and Pseudomonas aeruginosa were treated with chemicals in order to obtain specific pathogen free animals. Hamsters kept in the laminar flow rack were treated orally with metronidazole several times to obtain a flagellate-free colony. After all flagellates had been eradicated, one pair of animals were kept in an isolator and mating was allowed to occur. When their offspring reached the age of seven weeks, they were intramuscularly injected daily with netilmicin sulfate for 10 consecutive days. Following these treatments, all of the hamsters were free of Pasteurella and Pseudomonas. Further breeding of these animals was continued in isolators. To confirm the absence of selected pathogens, they were placed in a barrier room for further breeding as specific pathogen free animals.


Synergistic effects of interleukin 3 and interleukin 11 on murine megakaryopoiesis in serum-free culture. We investigated the effects of interleukin 11 (IL-11) on murine megakaryopoiesis in serum-free cultures, using nonadherent, nonphagocytic, and T-cell-depleted bone marrow cells. IL-11 alone had no influence on megakaryocyte (Meg) colony formation in serum-free methylcellulose cultures, but it significantly enhanced the growth of Meg and granulocyte-macrophage-Meg colonies supported by optimal and suboptimal concentrations of interleukin 3 (IL-3). IL-11 also increased the size of IL-3-dependent Meg colonies as well as increasing the size and DNA content of constituent Meg. In liquid cultures, IL-11 alone did not increase the number of Meg, but it enhanced their size and acetylcholinesterase (AchE) levels. The addition of IL-11 to cultures containing suboptimal concentrations of IL-3 resulted in a synergistic increase of Meg AchE. These results suggest that IL-11, similarly to interleukin 6, has an effect on Meg and acts synergistically with IL-3 to augment murine megakaryopoiesis in vitro.


Dual role of IL-7 in the growth and differentiation of immature thymocytes. The effects of interleukin 7 (IL-7) on subpopulations of CD4-CD8- thymocytes from young adult mice were tested in vitro. When highly purified CD3-CD4-CD8-thymocytes were cultured in the presence of recombinant IL-7, significant proportions of them became CD4+ and/or CD8+ within a day. CD3+ cells were also detected after 2 days. CD3-CD4-CD8- thymocytes were further subdivided into interleukin 2 receptor (IL-2R)- and IL-2R+ populations. The majority of the IL-2R- cells became CD4+ and/or CD8+ in 1 day in the presence of IL-7, and a substantial proportion of them also became CD3+ in 2-3 days. No significant number of CD4+ or CD8+ cells were generated from the IL-2R+ population under the same conditions. However, a small but significant proportion of them became CD3+ in 3-day cultures with IL-7. Although CD4+/CD8+ cells were also generated from the IL-2R- population in 1-day cultures in the absence of IL-7, the viability of the cells declined rapidly, and no significant numbers of CD3+ cells were generated. In proliferation assays, IL-7 alone vigorously stimulated relatively minor subpopulations of CD4-CD8- thymocytes. The IL-7-responsive cells were CD3+, did not express the IL-2R or the heat-stable antigen M1/69, and included both T-cell receptor (TCR)alpha beta + and TCR alpha beta- populations, the latter most likely TCR gamma delta +. The CD3+CD4-CD8- thymocytes, stimulated with IL-7 for 3 days, remained CD4-CD8-. These results demonstrate important roles of IL-7 in the growth and differentiation of CD4-CD8- thymocytes in vitro. It functions as a survival factor and allows CD3-CD4-CD8- cells to undergo their precommitted differentiation without inducing their proliferation, and it also stimulates CD3+CD4-CD8-thymocytes to proliferate without inducing their differentiation.


Lp(a) and the risk of coronary heart disease. A screening study was performed on 106 children with familial risk for coronary heart disease (CHD) and on matched controls. The two groups differed in several parameters. Children of CHD patients exhibited significantly elevated levels of Lp(a) and total cholesterol, reduced HDL apo A1 and apo A2 and increased values of serum hexuronic acid. These results support the concept that genetic and familial factors contribute to the risk of atherosclerosis.


Lp(a): a link between thrombosis and atherosclerosis. Lipoprotein(a), or Lp(a), is a lipoprotein having lipid composition similar to that of LDL, but a protein moiety consisting of ApoB 100 linked by disulfide bridge to apo(a), a glycoprotein with structural similarity to plasminogen. Lp(a) can be both atherogenic and thrombogenic. These two actions are likely to be mutually operative, a fact that on a molar basis makes Lp(a) more of a cardiovascular pathogen than LDL.


Fibrinogen/fibrin in atherogenesis.

Fibrinogen/fibrin in atherogenesis. Fibrin is a major component of many atherosclerotic plaques. Within the intima there is continuous formation of fibrin, and continuous fibrinolysis. In aortic lesions, a lipoprotein bound to fibrin can be released by incubation with plasmin. Most of this lipoprotein is accounted for by Lp(a). The atherogenicity of Lp(a) may be more associated with lipid deposition than with inhibition of fibrinolysis. Fibrin degradation products may be chemotactic to monocyte-macrophages and stimulate smooth muscle cell proliferation.


Disruption of ovarian development in alligator embryos treated with an aromatase inhibitor. It has been suggested that sex differentiation in vertebrates is steroid hormone dependent, that estrogens play a critical role in ovarian differentiation, and that male sex differentiation will occur in the absence of estrogens. Using the model of the alligator in which sex can be manipulated by incubation conditions (eggs incubated at a constant temperature of 30 degrees produce 100% females, and at 33 degrees produce 100% males), a series of experiments using antiestrogens, antiandrogen, estradiol-17 beta, dihydrotestosterone (DHT), and aromatase inhibitors were performed. Test substances were injected into alligator eggs prior to gonadal sex differentiation and the eggs were incubated at male or female temperatures until just before expected date of hatching. Gonads were excised and the sex was verified histologically. Control embryos injected with vehicle produced the expected sex: females at 30 degrees and males at 33 degrees. Estradiol in eggs at 33 degrees (male temperature) produced 100% females and did not alter female development in eggs at 30 degrees. Antiandrogen, DHT, and a steroid antiestrogen had no discernible effect in either the 30 degrees or the 33 degrees eggs at the doses tested. The aromatase inhibitors aminoglutethimide and 4-hydroxyandrostenedione caused a moderate disruption of ovarian development and had no apparent effect on testicular development. The nonsteroidal aromatase inhibitor, Ciba Geigy 16949A, caused inhibition of ovarian development in all treated embryos. The Mullerian ducts did not appear to be affected by this treatment, or by any of the other treatments, and the gonads did not appear masculinized. We conclude that estrogen appears to be necessary for normal ovarian development, but that inhibition of estrogen synthesis alone is insufficient to cause masculinization. Likewise, exogenous androgens appear unable to masculinize embryonic gonads. The evidence suggests that testicular differentiation in amniote vertebrates is dependent on factors other than androgens or level of estrogens.


Fetal atrial measurements before and after delivery: correlation with plasma atrial natriuretic peptide. Fetal and early neonatal atrial measurements correlating with plasma atrial natriuretic peptide were studied to evaluate the circulatory change before and after delivery in a longitudinal study of 10 normal fetuses from 1 week before until 5 days after delivery. Before delivery, right atrial area value was significantly larger than that of left atrial area; however, there were no significant differences between left and right atrial areas after delivery. Right atrial area value before delivery was significantly larger than those after delivery, whereas the left atrial area did not change before and after delivery. Combined atrial area value before delivery was also larger than those after delivery. Plasma atrial natriuretic peptide values did not change after delivery. These results suggest that the change from fetal to neonatal circulation mainly caused by the pulmonary circulation induces the change of right atrial size before and after delivery.


Role of saccharides in immunoglobulin--Fc receptor interactions. The rosettes formed by mouse peritoneal macrophages or DCH-5 cells and TNP-erythrocytes coated with anti-TNP antibodies of different isotypes were inhibited to various extent by monosaccharides. The most effective inhibitors were N-acetylglucosamine, glucosamine, mannose and N-acetylneuraminic acid in 1-5 mmol/L concentrations. Even more efficient were glycopeptides isolated from IgG molecules. The Fc receptors (FcRs) released from DCH-5 cells during cultivation and gradually separated by affinity chromatography on immobilized IgG reacted with aggregated IgG and inhibited the rosette formation. The FcRs eluted by monosaccharides influenced mainly the number of rosettes mediated by IgA and IgE while those eluted with a glycine-HCl buffer inhibited preferentially IgG rosettes. As shown by SDS-PAGE the heterogeneity of the fraction eluted with a mixture of monosaccharides revealed one main component with an effective molar mass of 50 kg/mol. The glycine-HCl eluate contained two major components of 55 and 38 kg/mol. The IgG-Sepharose 4B bound all the fractions but only the binding of the 50 kg/mol molecule could be inhibited by monosaccharides.


Abscisic acid and its synthetic analog in relation to growth and nitrogenase activity of Azotobacter chroococcum and Nostoc muscorum. The plant hormone abscisic acid as well as its synthetic analog LAB 173711 significantly increased the nitrogenase activity of the bacterium Azotobacter chroococcum and the cyanobacterium Nostoc muscorum. The effect depended on the concentration of the substances (0.001-10 mg/L) and the age of the cultures. The biomass of the organisms was not significantly influenced.


Prenatal diagnosis of bilirubin-UDP-glucuronosyltransferase deficiency in rats by genomic DNA analysis. Hepatic bilirubin excretion requires UDP-glucuronosyltransferase-mediated glucuronidation. Patients with type I Crigler-Najjar syndrome and mutant rats (Gunn strain) inherit deficiency of UDP-glucuronyltransferase activity toward bilirubin as an autosomal recessive trait and, as a result, exhibit marked nonhemolytic unconjugated hyperbilirubinemia throughout postnatal life. Heterozygous carriers of the trait have normal serum bilirubin levels. Because of placental excretion of unconjugated bilirubin, type 1 Crigler-Najjar syndrome patients and Gunn rats are not jaundiced in utero, making prenatal diagnosis difficult. Here we report a diagnostic method in Gunn rats based on genomic DNA analysis for prenatal recognition of deficiency of UDP-glucuronyltransferase activity toward bilirubin in Gunn rats and identification of heterozygous carriers. We and others have shown that two distinct messenger RNA species (UDP-glucuronyltransferase activity toward bilirubin and the 3-methylcholanthrene-inducible phenol-UDP-glucuronyltransferase messenger RNA) in Gunn rat liver contain identical deletions of a single guanosine residue in their common 3' regions. Loss of the restriction site for the endonuclease BstNI, which results from this deletion, was used as the basis for a diagnostic test. Female heterozygous Gunn rats were mated with male homozygous Gunn rats. Genomic DNA was extracted from the chorionic aspect of placenta of 17-day fetuses or from leukocytes from normal rats, obligate heterozygotes and homozygous Gunn rats. The DNA was sequentially digested with the restriction enzymes EcoRI and BstNI and subjected to Southern-blot analysis with a double-stranded DNA probe for the common region of UDP-glucuronyltransferase activity toward bilirubin and the 3-methylcholanthrene-inducible UDP-glucuronyltransferase messenger RNAs.(ABSTRACT TRUNCATED AT 250 WORDS)


Immediate allergy to natural latex: clinical and immunological studies. Since 1979 several reports of contact urticaria due to natural latex have been documented. In recent years cases of anaphylaxis, rhinitis, and asthma due to latex have appeared. Nine patients, studied in our clinic between 1986-1991, suffered immediate allergic reactions caused by rubber products. All showed an immediate skin reaction to latex extract. Rub testing with surgical gloves was positive in eight patients. Immunological techniques (RAST, ELISA, HRT) demonstrated specific IgE against latex. Specific bronchial provocation testing was performed in one patient who presented with asthma when she used latex surgical gloves. Patch testing to common rubber additives were negative in our patients. These results suggest that natural latex antigens present in rubber objects can cause hypersensitivity reactions probably due to IgE-mediated mechanisms.


Role of the liver in splanchnic extraction of atrial natriuretic factor in the rat. Mesenteric, hepatic and splanchnic extraction of C-terminal and N-terminal atrial natriuretic factor was investigated in male Sprague-Dawley rats. Plasma concentrations (mean +/- S.E.M.) of C-terminal atrial natriuretic factor were 55.0 +/- 6.1 fmol/ml, 31.2 +/- 4.0 fmol/ml and 23.5 +/- 3.3 fmol/ml (n = 12) in the abdominal aorta, the portal vein and the hepatic vein, respectively. N-terminal atrial natriuretic factor plasma levels in these vessels were 3031 +/- 756 fmol/ml, 2264 +/- 661 fmol/ml and 1618 +/- 496 fmol/ml (n = 6), respectively. Although the mesenteric extraction ratio was higher (p less than 0.05) for C-terminal atrial natriuretic factor (42% +/- 6%) than for N-terminal atrial natriuretic factor (28% +/- 4%), there were no significant differences in the hepatic extraction ratio (41% +/- 5% vs. 39% +/- 6%) and the splanchnic extraction ratio (56% +/- 5% vs. 50% +/- 7%). These data suggest a major role of the liver in the splanchnic extraction of C-terminal and of N-terminal atrial natriuretic factor in the rat.


Expression of murine cyclin B1 mRNAs and genetic mapping of related genomic sequences. Two cDNAs that encode a protein with 87% identity to human cyclin B1 and that differ only in the length of their 3'-untranslated regions have been isolated from a 70Z/3B murine pre-B leukemia cell library. Three sizes of RNA transcripts were detected in Northern hybridization analyses of a variety of normal tissues and transformed cell lines using the cDNA inserts as probes. The expression of these RNAs can be modulated in tissue culture cell lines by physiologically relevant stimuli, increasing when cells are stimulated to proliferate and decreasing when cells are induced to differentiate. Moreover, RNAs from tissues that contain few proliferating cells have no detectable hybridizing transcripts. The coordinate regulation of these RNAs with other genes that are activated during the cell division cycle and the profound similarity of the predicted amino acid sequence to those of published cyclin B homologues indicate that these genes encode a murine cyclin B1. In Southern hybridization analysis of BALB/cAnPt genomic DNA digested with EcoRI, 12 fragments hybridized with the cDNA probes. Through Southern blot analyses of DNA from backcross and cogenic mice, recombinant inbred strains, and somatic cell hybrids, the genetic loci that produce the cyclin B1-related sequences (designated loci Cycb1-rs1 to Cycb1-rs9) were mapped on mouse chromosomes 5, 1, 17, 4, 14, 13, 7, X, and 8, respectively. Cycb1-rs6 (on chromosome 13) is discussed as the most likely candidate for an expressed structural gene locus.


Radiation hybrid map spanning the Huntington disease gene region of chromosome 4. Radiation hybrid (RH) mapping was used to construct a map of 11 markers in the distal 4 Mb of the short arm of chromosome 4, the region containing the Huntington disease gene. Two different methods for deriving the order of the markers were compared and both arrived at the same order as being the most likely. This order is also consistent with both the physical map constructed using pulsed-field gel electrophoresis (PFGE) and the meiotic linkage map. Comparing the RH map to the map determined by PFGE provided the means to equate RH map units (centirays) with actual physical distance in kilobases of DNA. In addition, a simple procedure for reducing the complexity of human DNA in radiation hybrids is described. One cell line isolated using this procedure contains, as its only human DNA, approximately 2 Mb surrounding the Huntington disease gene.


An estimate of the number of genes in the Huntington disease gene region and the identification of 13 transcripts in the 4p16.3 segment. Physical mapping and genetic linkage studies have positioned the Huntington disease (HD) gene to a relatively large genomic region in the distal portion of the short arm of human chromosome 4 (4p16.3). To estimate the number of genes present in this region and to identify candidate disease genes, several clones that map to the 4p16.3 segment have been examined for clusters of CpG-rich restriction sites and transcribed sequences. Thirteen expressed sequences were identified and were shown by pulsed-field gel electrophoresis not to cluster into a small segment of the 4p16.3 band. The frequency of transcripts in these clones suggests that the putative HD gene region contains about 100 genes.


Chromosomal localization of three human D5 dopamine receptor genes. It is currently thought that genetic predisposition to imbalances in dopaminergic transmission may underlie several neurological disorders, including schizophrenia, manic depression, Tourette syndrome, Parkinson disease, Huntington disease, and alcohol abuse. Originally two receptors, D1 and D2, were thought to account for all of the pharmacological actions of dopamine. However, through homology screening three additional genes, D3, D4, and D5, and two pseudogenes closely related to D5 have been characterized. To begin our genomic and evolutionary analyses of the human D5 dopamine receptor gene and its two pseudogenes, we have mapped each of them to their respective chromosomes. By combining in situ hybridization results with sequence analysis of PCR products from microdissected chromosomes, somatic cell hybrids, and radiation hybrids, we have assigned DRD5 (the locus containing the functional human D5 receptor gene) to chromosome 4p16.1, DRD5P1 (the locus containing D5 pseudogene 1) to chromosome 2p11.1-p11.2, and DRD5P2 (the locus of D5 pseudogene 2) to chromosome 1q21.1.


The mouse CREB (cAMP responsive element binding protein) gene: structure, promoter analysis, and chromosomal localization. In this paper we report the isolation and characterization of the mouse CREB gene. It is composed of 11 exons and 10 introns and spans a region of 70 kb. BR-A and BR-B, the two alpha-helical regions of the proposed basic DNA binding domain of CREB, are encoded separately on exons 10 and 11. The mouse CREB gene is expressed from a promoter that is situated in a CpG island. The promoter contains no TATA or CCAAT box homologies but has a number of putative binding sites for the acidic transcriptional activator Sp1 and a 9/11 match with the initiator region. Transcriptional start site mapping identified five major start sites spread over at least 41 nucleotides. Northern blot analysis indicated that expression of the CREB gene is almost ubiquitous with expression at differing levels of multiple transcripts. Testis expressed a predominant RNA species of approximately 1.6 kb. The CREB gene was found to be single copy in the mouse and well conserved through evolution. Finally Creb-1, the CREB locus, was mapped to the proximal region of mouse chromosome 1.


Effects of prolactin and androgens on enzymes of carbohydrate metabolism in prostate of castrated bonnet monkeys Macaca radiata (Geoffroy). Effects of prolactin (PRL), bromocriptine (Br), testosterone propionate (TP), dihydrotestosterone (DHT) and the combinations of these androgens with PRL/Br on the specific activities of caudal and cranial prostatic cellular enzymes involved in carbohydrate metabolism in castrated mature bonnet monkeys have been studied. Castration decreased all the enzymes studied such as hexokinase (HK), 6-phosphofructokinase (6-PFK), glyceraldehyde-3-phosphate dehydrogenase (G-3-PD), pyruvate kinase (PK), glucose-6-phosphate dehydrogenase (G-6-PD) and 6-phosphogluconate dehydrogenase (6-PGD) in the cranial and caudal prostates. PRL elevated the activities of all the enzymes above normal except G-3-PD of cranial lobe. In the caudal lobe, PRL brought back the activities of HK, PFK, PK, G-6-PD to normal and 6-PGD above normal except G-3-PD. TP/DHT treatment increased all the enzymes in both the lobes. PRL given along with TP/DHT further enhanced the androgen action with regard to HK, PK, G-6-PD and 6-PGD of cranial and PFK, G-3-PD, PK, G-6-PD and 6-PGD of caudal lobe. Br treatment did not produce any alteration of these enzymes in both the lobes. In the cranial lobe, during Br+TP/DHT treatment, the stimulating effects of androgen were unaffected on all the enzymes except PK. On the other hand in the caudal, the stimulatory effects of androgens were affected and the activities of HK, PFK, PK and 6-PGD were significantly decreased. The present results suggest that PRL has a direct as well as a synergistic action with androgens on enzymes of EMP and HMP shunt in the prostates of monkeys.


Cutaneous adverse reactions following the administration of nonsteroidal antiinflammatory drugs and antibiotics: an Italian survey. The Italian Group for Epidemiological Research in Dermatology (GISED) collected a series of cutaneous adverse reactions following NSAID and/or antibiotics administered by topical and/or systemic route. Dermatologists from North and Central Italy took part in this survey by filling in 1457 case report forms during a four-month observation time in 1988-89. The main purpose of our epidemiological study aimed at evaluating a post-marketing surveillance program by examining spontaneous reports of cutaneous adverse reactions. This result seems to be noteworthy, considering the difficulties encountered in Italy to develop such a program.


Laparoscopic common bile duct exploration. Laparoscopic cholecystectomy is now the standard approach to gallbladder disease. While laparoscopic cholecystectomy offers many advantages over the conventional laparotomy procedure one of its drawbacks is that a synchronous common bile duct exploration, for so long a cornerstone of management of choledocholithiasis, is not yet widely practised laparoscopically. Endoscopic sphincterotomy or open surgery, with their attendant hazards and morbidity, remain the most common approaches. A flexible choledochoscope with an operating channel may in future facilitate laparoscopic management of choledocholithiasis but as yet this is not widely available. We report the removal of a common duct stone by dormia basket extraction through the cystic duct at laparoscopic cholecystectomy.


Doxazosin in the management of hypertensive diabetes--a cautionary note (?). Twenty-two patients with diabetes and hypertension were treated with Doxazosin. An acceptable fall in blood pressure was found with 1 mg. in 50% of patients and 2-8 mgs. in 50%. An increase in HDL cholesterol and a fall in LDL cholesterol levels which reached statistical significance was observed. A small but significant increase in HBA1 levels occurred in the 50% of patients on the higher Doxazosin dose.


Prevalence of disabilities in a national sample of 7-year-old Israeli children. The prevalence of chronic conditions and illnesses causing disability in Israeli Jewish children aged 7 years born in 1975 was studied on the basis of a national sample (n = 7,739). Eighty medical conditions causing disability were defined. The study showed a total disability rate of 17.5%, which is higher than that reported on a similar national sample of 3 year olds (prevalence of 6.9%). The percentage of disabilities among very low birthweight children and those with family problems was four times greater than in the total population. Mild retardation and undefined learning problems were more prevalent among children of mothers with low educational level and among children whose birth order was fourth or more. Asthma and spastic bronchitis were more prevalent among children whose mothers were of European/American origin (P less than 0.05). Behavior and mental disorders, learning problems, speech and language disorders were more prevalent among male children. Two-thirds of the children with a diagnosed problem also had at least one functional disability. There were relatively more children from lower social classes in the special education schools than in the national sample. Increased prevalence of disabilities among children of very low birthweight, low maternal educational level, high birth order, and those from families whose origin is Asian/African and from families with intrafamilial problems defines the children at risk for disabilities and placement in special education schools.


[Quality control in epicutaneous testing--reproducibility in the right-left comparison]. In 121 patients, 26 substances of a standard test battery were applied symmetrically on both sides of the upper back. The Finn chamber on Scanpor was the test system used, the final reading being taken on day 3 or 4. Of 166 tests, 131 were positive on both sides. Test substances and patients age had no influence. In comparison with the results in the literature, ours show a good reproducibility of 78.9%. Doubtful reactions (?+) were difficult to evaluate. These reactions deserve future research.


[Lichenoid eruptions after gold therapy. Report of two cases]. Lichenoid eruptions appearing in two women receiving chrysotherapy (Solganal B oleosum R) a described. The morphological diagnosis, distribution of skin eruptions, histological and immunological features and monoclonal antibodies (Leu 4, Leu 2A and Leu 3A) against several cell surface markers were studied. Individual lesions were not distinguishable from idiopathic lichen planus, with some features typical for gold lichenoid eruptions: hyperpigmentation, psoriasiform eruptions, and hair loss. Histopathological and immunological examination of biopsy specimens with monoclonal antibodies generally confirms clinical impressions. The authors suggest similar pathogenetic mechanisms in the development of idiopathic lichen planus and of lichenoid eruptions after gold therapy.


The health of mothers and fathers with a child with a disability. Iris Cairns examined the different effects on the health of mothers and fathers when they have a child with a disability. Both have a higher incidence of psychiatric morbidity than parents in the general population, with the mothers' health more significantly affected than that of the fathers, her study revealed.


Quantitative determination of calcium-activated myosin adenosine triphosphatase activity in rat skeletal muscle fibres. A quantitative histochemical technique was developed for determining the kinetics of the calcium-activated myosin ATPase (Ca(2+)-myosin ATPase) reaction in rat skeletal muscle fibres. Using this technique, the maximum velocity (Vmax) and the apparent Michaelis-Menten rate constant for ATP (K(app)) of the Ca(2+)-myosin ATPase reaction were measured in type-identified fibres of the rat medial gastrocnemius (MG) muscle. The Vmax and the K(app) of the Ca(2+)-myosin ATPase reaction were lowest in type I fibres and highest (i.e., approx. two times greater) in type IIb fibres. The K(app) in type IIa fibres was similar to that in type I. However, the Vmax was 1.5 times greater in type IIa fibres, compared to type I fibres. Evidence is presented to suggest that the type IIb fibre population in the MG does not represent a single myosin isozyme. In addition, the broad range of Vmax and K(app) values indicates that there is marked heterogeneity in the myosin heavy chain and myosin light chain composition of myosin isozymes among individual fibres.


Use of etretinate for treatment of primary keratinization disorders (idiopathic seborrhea) in cocker spaniels, west highland white terriers, and basset hounds. An open clinical trial was used to evaluate the synthetic retinoid, etretinate, for treatment of idiopathic seborrhea in Cocker Spaniels, West Highland White Terriers, and Basset Hounds. Clinical and histologic improvement was seen in the Cocker Spaniels. Etretinate had no beneficial effect on the skin disease of the West Highland White Terriers or the Basset Hounds. Etretinate treatment did not alter the type or degree of otitis externa. Clinical side effects were minimal. Relevant laboratory abnormalities were not detected.


In vivo antitumor effects of fluoropyrimidines on colon adenocarcinoma 38 and enhancement by leucovorin. Antitumor effect and active metabolites of fluoropyrimidines were examined in mice with transplantable colon adenocarcinoma 38 (Co 38). 5-Fluoro-2'-deoxyuridine (FUdR) treatment resulted in a much higher level of free 5-fluoro-2'-deoxyuridine-5'-monophosphate in the tumor than 5-fluorouracil (5-FU) did, and thymidylate synthase was almost completely inhibited after FUdR treatment, but FUdR showed weaker antitumor activity than 5-FU did. Moreover, 5-fluorouridine (FUR) also hardly inhibited tumor growth. A more marked tumor inhibition was obtained when FUdR and FUR were administered together. The antitumor activity of 5-FU was similar to that of the combination of FUdR and FUR. In combination with 2,2'-anhydro-5-ethyluridine, a uridine phosphorylase inhibitor, FUdR lost its antitumor activity, but that of FUR was somewhat potentiated. On the other hand, in combination with leucovorin (LV), 5-FU showed markedly potentiated antitumor activity, while the antitumor activity of FUdR or FUR was not potentiated. Addition of LV to the combination of FUdR and FUR enhanced the inhibitory effect of the drugs. From these results, the combination of FUdR and FUR together with LV, and the combination of 5-FU and LV seem to be highly efficacious against Co 38.


Fast and slow intrafusal fibre type systems in chicken leg muscle spindles. The results of this study indicate that in serial sections incubated with monoclonal antibodies against MHC and stained for mATPase, intrafusal fibres in chicken leg muscle spindles are separable into slow and fast types. As observed here, fast fibres are a homogeneous group; however, it had been shown earlier (Maier & Zak, 1990) that based on relative amounts of slow MHC, the slow fibres can be further divided into 2 subgroups. These 3 types of intrafusal fibre conform to 3 recently demonstrated patterns of motor innervation (Maier, 1991). It is suggested that the respective actions of slow and fast intrafusal fibres produce different components of the afferent discharge of chicken muscle spindles.


Myosin heavy chain expression in rabbit masseter muscle during postnatal development. The expression of isoforms of myosin heavy chain (MHC) during postnatal development was studied in the masseter muscle of the rabbit. Evidence is presented that in addition to adult fast and slow myosin, the rabbit masseter contains neonatal and 'cardiac' alpha-MHC. During postnatal growth myosin transitions take place from neonatal and fast (IIA, IIA/IIB--referring to a fibre containing both IIA and IIB MHCs) MHC to adult 'cardiac' alpha-MHC and I/alpha-MHC. Since there is a temporary population of fibres containing IIA/alpha-MHC during the first 4 wk of development with a peak in the 3rd to 4th wk, the transition from IIA-MHC to alpha-MHC may occur in these IIA/alpha-MHC-containing fibres. The appearance of 'cardiac' alpha-MHC coincides with the timing of weaning, suggesting that the changes in MHC content, that probably result in a transition to a lower speed of contraction, have functional significance related to weaning. The finding of neonatal MHC in adult rabbits indicates that the masseter develops at a rate and in a way that is distinct from most other skeletal muscles. A spatiotemporal variation in expression of myosin isozymes within the masseter was observed, with many fibres containing more than one myosin type, indicating developmentally regulated spatial differences in function.


Individual efficacy of clarithromycin (A-56268) and its major human metabolite 14-hydroxy clarithromycin (A-62671) in experimental pneumococcal pneumonia in the mouse. In man, clarithromycin (A-56268) is metabolized to a 14-hydroxy derivative (14-OH clarithromycin, A-62671); this metabolite is absent in mice. An experimental mouse model was used to compare the efficacy of clarithromycin versus 14-OH clarithromycin in pneumococcal pneumonia by treatment with either the parent drug or its 14-OH metabolite alone. Four groups of 15 mice were infected intra-tracheally with a virulent strain of Streptococcus pneumoniae serotype 3 (approximately 10(5) cfu/mouse). Treatment was begun 18 h post infection, with clarithromycin or erythromycin 50 mg/kg subcutaneously (sc) bd, 14-OH clarithromycin 25 mg/kg sc bd or antibiotic free saline sc bd for 24 or 48 h. Survival rates ten days after one day of treatment with clarithromycin, 14-OH clarithromycin, erythromycin, or saline were 53%, 60%, 27% and 0% respectively. After two days treatment the rates were 100%, 100%, 53% (P less than 0.01) and 0% respectively. Thirty-six h after two doses, 14-OH clarithromycin demonstrated superior intra-pulmonary killing activity to erythromycin (2.7 +/- 0.4 vs 6.6 +/- 0.8 log10 cfu/mL lung homogenate) (P less than 0.001) and comparable activity to clarithromycin (3.6 +/- 1.3 log10 cfu/mL). These results show that clarithromycin (50 mg/kg sc) and 14-OH clarithromycin (25 mg/kg sc) have similar in vivo efficacy in a mouse model, in which the 14-OH metabolite is not produced from clarithromycin. This suggests that in man, the clinical efficacy of clarithromycin is not impaired by metabolism to 14-OH clarithromycin, which achieves serum concentrations in man of approximately 1 mg/L after multiple doses of clarithromycin 500 mg po.


TLC G-65 in combination with other agents in the therapy of Mycobacterium avium infection in beige mice. The activity of TLC G-65 (a liposomal gentamicin preparation), alone and in combination with rifapentine, clarithromycin, clofazimine and ethambutol, was evaluated in the beige mouse (C57BL/6J--bgj/bgj) model of disseminated Mycobacterium avium infection. TLC G-65 was found to be more active than amikacin. The combination of rifapentine and TLC G-65 was more active than either agent alone. The activity of clarithromycin in combination with TLC G-65 was similar to that of either agent alone. Clofazimine improved the activity of TLC G-65 with respect to the spleen, while ethambutol improved the activity with respect to the liver. Clofazimine and ethambutol enhanced the activity of TLC G-65 against bacteria in the lungs. TLC G-65 in combination with rifapentine appears to be an attractive regimen for the treatment of infections caused by bacteria in the M. avium complex.


Immunocytochemical localization of annexin V (CaBP33), a Ca(2+)-dependent phospholipid- and membrane-binding protein, in the rat nervous system and skeletal muscles and in the porcine heart. We investigated the ultrastructural localization of annexin V a Ca(2+)-dependent phospholipid- and membrane-binding protein in the nervous system, heart, and skeletal muscles. The results indicate that in the cerebellum the protein is restricted to glial cells, where it is found diffusely in the cytoplasm as well as associated with plasma membranes. Bergmann glial cell bodies and processes and astrocytes in the cerebellar cortex and oligodendrocytes in the cerebellar white matter displayed an intense immune reaction product. In sciatic nerves, the protein was exclusively found in Schwann cells with a subcellular localization similar to that seen in glial cells in the cerebellum. Pituicytes in the neurohypophysis were intensely immunostained, whereas axons were not. In the heart, annexin V was restricted to the sarcolemma, transverse tubules, and intercalated discs. In skeletal muscles the protein was localized to the sarcolemma and transverse tubules. No evidence for the presence of the protein in the sarcoplasm or in association with mitochondria, the sarcoplasmic reticulum, or contractile elements was obtained. The observation that plasma membranes in cells expressing annexin V have the protein associated with them is in agreement with previous data on Ca(2+)-dependent binding of the protein to brain and heart membranes, and on existence of both EGTA- and Triton X-100-extractable and resistant fractions of annexin V in these membranes. The present data support the hypothesis that annexin V might be involved in membrane trafficking and suggest a role for this protein in the regulation of cytoplasmic activities in glial cells.


Influence of long-term hypoxia on the energy metabolism of the haemoglobin-containing bivalve Scapharca inaequivalvis: critical O2 levels for metabolic depression. The oxygen consumption rate of Scapharca inaequivalvis measured under normoxic conditions over 48 h showed a significant daily cycle with lowest values occurring shortly after the dark period; all hypoxia exposure experiments were carried out during the declining part of the cycle. Animals were exposed to a constant level of hypoxia for a 12-h period in a series of 14 experiments, each at a different oxygen tension. The oxygen consumption was measured continuously, and the extent of accumulation of end-products (succinate and propionate), and the inhibitory effect of adenosine triphosphate on phosphofructokinase were determined at the end of exposures. All three parameters (oxygen consumption, end-product accumulation, phosphofructokinase inhibition) showed a remarkable correlation with major changes occurring between 2.5 and 1.5 ppm (7 and 4 kPa) O2. The oxygen consumption rates showed a drop to 6% of the normoxic rate, but a consistent low consumption remained below 2 ppm (5.5 kPa) which partly recovered over the 12-h exposure period by about three-fold. Succinate and propionate accumulated progressively between 2.5 and 1.5 ppm (7 and 4 kPa); at [O2] less than 1.5 ppm (4 kPa) the concentration did not increase further, indicating that anaerobic metabolism had reached a maximum. Over the same range, phosphofructokinase showed an increased sensitivity for adenosine triphosphate, the lower inhibitor concentration at 50% Vmax value pointing to depression of glycolytic rate. Despite the activation of anaerobic metabolism and the evident depression of aerobic metabolism, simple calculation demonstrates that Scapharca inaequivalvis relies mainly on aerobic metabolism even during severe hypoxia.(ABSTRACT TRUNCATED AT 250 WORDS)


Influence of aldehydes on selected mechanical properties of resin composites. The present study investigated whether propanol, a monofunctional aldehyde, was able to improve the mechanical properties of dental polymers. The underlying hypothesis was that a cross-linking reaction is possible between various functional groups of different polymers. Propanol was added to monomer mixtures, which were then made light-curing and loaded with filler. The monomer mixtures were varied with respect to monomer composition and content of aldehyde. Four mechanical properties of the experimental resin composites were determined. Addition of propanol gave rise to significant improvements in mechanical properties, which may be indicative of a cross-linking ability of monofunctional aldehydes. With the exception of modulus of elasticity, the mechanical properties of resin composites based on UEDMA/HEMA were superior to those of BISGMA/TEGDMA-based materials, even though the improvements in flexural strength and modulus of resilience were most pronounced for the BISGMA/TEGDMA-based resin composites.


Plasma endothelin levels in cirrhotic subjects. Endothelin-1, a potent vasoconstrictor peptide with 21 amino acid residues, is released by the vascular endothelium. Plasma immunoreactive endothelin levels were measured in 23 patients with cirrhosis and in 20 healthy subjects. Concentrations were significantly lower in patients with non-uraemic cirrhosis than in normal subjects (19.4 +/- 8.9 pmol/l vs. 48.8 +/- 24.8 pmol/l, p less than 0.002). Plasma renin, aldosterone, atrial natriuretic peptide, arginine-vasopressin and catecholamines did not show significant correlations with plasma endothelin-1 levels. Furthermore, there were no significant differences in plasma endothelin levels for etiology of cirrhosis, presence of ascites or varices. These data suggest that low circulating endothelin may be involved in the development or maintenance of systemic vasodilatation in cirrhosis.


Ontogeny and distribution of the murine B cell Fc gamma RII. The distribution and expression of the IgG FcRII (Fc gamma RII) on normal murine B cells was examined. Using multicolor flow cytometry, spleens from neonatal mice of increasing age and adult bone marrow were analyzed for expression of the Fc gamma RII. In addition, B cells from peripheral lymphoid organs, as well as panel of B cell tumors, were tested. The results demonstrate that the Fc gamma RII is expressed on all pre-B cells and immature B cells in the neonatal spleen and adult bone marrow, on all mature B cells in peripheral lymphoid organs, and on switched B cells in Peyer's patches. Furthermore, the Fc gamma RII was found to be present on B cell tumors representative of all stages of B cell maturation and differentiation. Taken together, the results indicate that Fc gamma RII is expressed during the entire lifetime of the B cell. In addition, examination of spleen cells from neonatal mice revealed a large number of pre-B cells, phenotypically defined as B220+, IgM-. These pre-B cells were present at birth, peaked in number between 2 and 3 wk of age, and became a minor population by day 30. Further phenotypic analysis of these cells demonstrated the expression of the BLA-1 and BP-1 Ag, and the lack of T cell and NK cell markers, thus confirming their assignment to the B cell lineage. Finally, the Fc gamma RII present on these pre-B cells was shown to be functional, by virtue of its ability to bind aggregated IgG.


Analysis of an in vitro-generated signal that induces systemic immune deviation similar to that elicited by antigen injected into the anterior chamber of the eye. The selective deficit in delayed hypersensitivity that characterizes anterior chamber-associated immune deviation (ACAID) is the direct result of a blood borne, Ag-specific, cell-associated signal that is created after Ag is injected into the anterior chamber of the eye of normal mice. The cells that carry this signal via the blood to the spleen express the mature macrophage marker F4/80 and are similar to, or perhaps even arise from, F4/80+ dendritic cells found within the stroma of normal iris and ciliary body. We have recently reported that ACAID-inducing properties can be conferred upon conventional F4/80-bearing macrophages harvested from the normal peritoneal cavity by incubating these cells in vitro with the soluble protein Ag, BSA, in the presence of supernatants harvested from cultured iris and ciliary body cells. Using this in vitro induction system, we have examined the limiting conditions for conferring ACAID-inducing potential on peritoneal exudate cells. We have found that an ACAID-inducing signal can be created in vitro with several different soluble Ag, including the retinal autoantigen-interphotoreceptor retinol binding protein, and that active endocytosis and processing by peritoneal exudate cells is required because chloroquine prevents these cells from acquiring ACAID-inducing properties. In addition, we have determined that for supernatant-treated peritoneal macrophages to induce ACAID to soluble Ag the cells must be 1) alive, 2) injected i.v. or i.p. (but not s.c.), and 3) administered to recipients with an anatomically intact spleen. When these conditions are met, as few as 20 F4/80+ macrophages pulsed with Ag in the presence of iris and ciliary body supernatants are sufficient to induce ACAID. Macrophage hybridomas derived from "conventional" APC can acquire ACAID-inducing potential in vitro if exposed to iris and ciliary body supernatants, whereas macrophage hybridomas derived from "suppressor inducer" APC constitutively possess ACAID-induced potential. Peritoneal macrophages that were endowed with ACAID-inducing properties by in vitro exposure to supernatants were found to elicit splenic suppressor cells similar to those found in spleens of mice with ACAID. Moreover, the expression of experimental autoimmune uveitis in mice immunized with interphotoreceptor retinol binding protein was significantly suppressed if the animals were pretreated with peritoneal exudate cells pulsed with this Ag in the presence of iris and ciliary body supernatants.(ABSTRACT TRUNCATED AT 400 WORDS)


Cyclosporine A and peripheral tolerance. Inhibition of veto cell-mediated clonal deletion of postthymic precursor cytotoxic T lymphocytes. Veto cell-mediated suppression of CTL responses has been proposed as one mechanism by which self tolerance is maintained in mature T cell populations. We have reported that murine bone marrow cells cultured in the presence of high-dose IL-2 (activated bone marrow cells) mediate strong veto suppressor function in vitro and in vivo, and that such veto activity is effected through clonal deletion of cytotoxic T cell precursors. In our studies, we have determined that bone marrow cell populations from athymic NCr-nu mice (H-2d) mediate strong veto cell activity without exposure to exogenous IL-2 in vitro. To examine mechanisms by which these naturally occurring veto cell populations in BM suppress precursor CTL (pCTL) responses, we used as a responding cell population in MLC, spleen cells of transgenic mice expressing at high frequency TCR specific for H-2 Ld encoded Ag with stimulation by H-2d-expressing cells in culture. Flow cytometric analysis was performed by staining the responding MLC cell population with the mAb 1B2 specific for the transgene-encoded TCR and determined changes of 1B2+ T cells. Such experiments demonstrated that the anti-H-2d cytotoxic response by these cell populations was specifically suppressed by NCr-nu (H-2d) bone marrow, and that 1B2+ pCTL were in fact specifically deleted from the responding cell population by incubation with such naturally occurring veto cell populations expressing the appropriate target Ag. In addition, to further understand the interactions of pCTL and veto cells and possible contributions by the latter to peripheral tolerance, we evaluated the effect of cyclosporine A (CsA) on veto cell-mediated suppression of pCTL of the transgenic mice. CsA inhibited veto cell-mediated suppression of cytotoxic T cell responses, and this inhibition correlated with a lack of clonal deletion of pCTL by veto cells in the presence of CsA. Furthermore, CsA exerted its effect through pCTL and not through veto cells, indicating that pCTL may play an active role in their own deletion by veto cells.


Age-dependent increase of extrathymic T cells in the liver and their appearance in the periphery of older mice. The liver is a major site of generation of extrathymic T cells with unique properties (e.g., expressing intermediate TCR and containing self-reactive clones). We investigated herein whether the levels of extrathymic alpha beta T cells varied in various organs as a function of age. A systematic examination of the number of mononuclear cells in various organs of BALB/c mice revealed that the number of hepatic MNC increased with age whereas the number of thymocytes decreased. These changes were more striking in mice fed under conventional conditions than under specific pathogen-free condition. The age-dependent changes in the number of mononuclear cells in the spleen and lymph nodes were minimal. Although the total proportion of alpha beta T cells in each organ remained constant, the staining patterns of TCR-alpha beta as shown by immunofluorescence profiles varied. The most prominent change was that intermediate TCR-alpha beta cells, which constituted a small population in the liver of young mice, expanded in the liver of older mice. Intermediate TCR cells appeared even in the periphery of older mice. These findings were confirmed by the appearance of extrathymic T cells with other unique properties, e.g., double-negative CD4-8- phenotype and CD44 expression. In athymic nude mice, only intermediate TCR cells were present in the liver and periphery. An age-dependent increase of intermediate TCR cells was also seen in these mice. Taken together with the result of bromodeoxyuridine-injection experiment, which showed an intensive in vivo proliferation of cells in the hepatic sinusoids, extrathymic T cells may differentiate predominantly in the liver and appeared even to the periphery in older mice.


Induction of cytotoxic T lymphocytes in vivo with protein antigen entrapped in membranous vehicles. Intravenous administration of APC such as splenocytes loaded with a soluble protein Ag has been shown to prime for an Ag-specific CTL response. It is thought that the APC directly presents loaded Ag in a MHC-restricted manner. However, it is demonstrated in this study that allogeneic splenocytes, MHC-free RBC, and even synthetic lipid vesicles (liposomes) after loading with OVA can elicit an OVA-specific and MHC-restricted CTL response. Biodistribution studies of these Ag-associated vehicles showed that the liver, spleen, and lung were the major organs responsible to scavenge these carriers, suggesting that the monocyte-macrophage system was involved in the Ag presentation for CTL. Depletion of macrophages by a specific macrophage killer, Cl2MDP, containing liposomes, abolished the CTL induction by immunization with OVA Ag carried by these vehicles except the induction by syngeneic splenocytes. Thus, the syngeneic splenocytes present Ag directly to the T cells, but other membranous vehicles carry the Ag to the host APC including macrophages, which then present it to the T cells. These results indicate that formulation of an Ag in membranous/colloidal vehicles may be a way to prime for a CTL response.


T cell receptor expression can switch on and off at a posttranslational level. The human T acute lymphocytic leukemia cell line, SUP-T13, is a mosaic of TCR/CD3+ and TCR/CD3- cells. Individual SUP-T13 cells can spontaneously switch on and off surface TCR/CD3 expression. This switching was demonstrated by culturing and analysis of single cell clones that were TCR/CD3+ or TCR/CD3-. The rate of switching is about 10(-2)/cell per generation in either direction. This is too high to be due to a spontaneous mutation event. Furthermore, switched cells can revert at similar rates, as demonstrated by repeated cloning and reanalysis. This makes it likely that a regulatory change is responsible for switching. In support of this, all known TCR/CD3 proteins are found intracellularly in TCR/CD3- cells, and they associate with each other as in TCR/CD3+ cells. Furthermore, no structural abnormalities of the TCR/CD3 chains can be seen in TCR/CD3- cells using two-dimensional electrophoresis. However, in these cells, the chains accumulate in great excess intracellularly. This accumulation is specific to the TCR/CD3 complex, as other glycoproteins are still expressed normally on the cell surface. Thus, there is regulation of TCR expression at a posttranslational level. These TCR/CD3- cells may lead to the identification of novel protein(s) involved in glycosylating, processing, or transporting the TCR/CD3 complex. Potential loss of TCR/CD3 expression may also limit the feasibility of TCR-based therapies for T cell leukemias.


Direct involvement of the CDR3-like domain of CD4 in T helper cell activation. The CD4 glycoprotein, a member of the Ig super-family, has long been known to play an important role in the immunologic activation of Th cells. The precise manner in which CD4 participates in this activation process is not yet understood. In an attempt to further define its role in Th cell activation, we modeled the D1 domain of the murine CD4 protein (L3T4) based on the experimentally determined high resolution structure of the human CD4 protein. Because the D1 domain of CD4 strongly resembles the V kappa chain of an antibody, we addressed the question of whether the CDR-like regions of CD4 are also involved in mediating protein-protein interactions. Consequently, we used the modeled L3T4 structure as a template in the design of conformational mimics of the CDR3-like region (residues 86-94). Only the analog designed to mimic both the sequence and conformation of this region exhibited highly specific inhibition of CD4-dependent responses. Because the inhibitory activity could be localized to the Th cell itself, it appears that this analog acts by uncoupling a CD4 association (independent of an APC) critical to generating a proliferative response.


IL 7-induced T cell receptor-gamma gene expression by pre-T cells in murine fetal liver cultures. IL-7 has previously been shown to stimulate proliferation of immature CD4-CD8- thymocytes from fetal and adult mice. We show that IL-7 mRNA levels are much higher in the day-14 fetal thymus than in the adult thymus, i.e., when the thymus is primarily composed of CD4-CD8- cells. Inasmuch as IL-7 may, therefore, be one of the first cytokines to which pre-T cells from the fetal liver are exposed when they migrate to the thymus during development, we hypothesized that IL-7 may promote differentiation of pre-T cells from the fetal liver. We found that human rIL-7 promotes growth of cells obtained from the liver of fetal mice at day 14 of gestation. Moreover, we found that IL-7 stimulated expression of mRNA encoding TCR-gamma, alpha-, and beta-genes. The size of the transcripts induced by IL-7 suggests that TCR-gamma was expressed in a rearranged form in response to IL-7, whereas TCR-alpha and -beta genes appeared to be expressed from nonrearranged loci. Culture of fetal liver cells for 7 days with IL-7 also caused the appearance of a population of cells expressing Thy-1 and Pgp-1 Ag, a phenotype similar to that seen in the early (day-14) fetal thymus. The Thy-1+ Pgp-1+ population was enriched in TCR-gamma mRNA, as determined by flow cytometric sorting followed by Northern blot analysis. These data support the hypothesis that IL-7 can stimulate some of the early events associated with thymocyte differentiation in the fetus.


Pattern of IL-1 receptor gene expression suggests role in noninflammatory processes. The cytokine IL-1 can be elaborated by most nucleated cell types and exerts its pleiotropic effects on a variety of tissues through binding to its cognate receptor. Two forms of IL-1R, designated types I and II, have been identified. These proteins share limited amino acid identity but both bind IL-1 alpha and IL-1 beta. A large number of cell types have been shown to possess IL-1R in vitro, but few studies have addressed the question of in vivo expression. Using in situ hybridization in normal adult BALB/c mice, we have examined the tissue distributions of both IL-1R types. The results demonstrate that although lymphoid tissues in healthy animals express low or nondetectable levels of receptor transcript, nonlymphoid tissues constitutively express readily detectable levels of IL-1R mRNA. Thymus and spleen samples were largely negative for both IL-1R types I and II, whereas half of the lymph nodes examined expressed IL-1R type I. In nonlymphoid tissues, IL-1R type I transcript was detected in the dentate gyrus of the brain, endocrine pancreas, cardiac endothelium, epidermis, dermis, hair follicle epithelium, uterine serosa, vaginal stroma, vaginal squamous epithelium, developing oocytes, and granulosa cells of ruptured follicles. In contrast, the IL-1R type II probe hybridized to epidermis, dermis, and vaginal basal epithelium. The two types of IL-1R were rarely, if ever, co-expressed by the same cell. The distributional segregation of the receptor types and the expression of IL-1R in normal nonlymphoid tissues has broad implications for the role of IL-1 in homeostatic physiology.


Tumor necrosis factor-alpha and IL-6 up-regulate IFN-gamma receptor gene expression in human monocytic THP-1 cells by transcriptional and post-transcriptional mechanisms. IFN-gamma is a potent activator of monocytic cell functions, including the stimulation of TNF-alpha and the control of IL-6 synthesis. These cytokines might act as autocrine or paracrine factors together with IFN-gamma in inducing biologic responses. In this study, we addressed the question of whether or not TNF-alpha and IL-6 play a role in modulating of IFN-gamma binding. Expression of IFN-gamma surface receptor was measured in the human monocytic THP-1 cells. The capacity of these cells to bind IFN-gamma increased with time after treatment with each cytokine. The number of IFN-gamma R per cell rose by three- and fourfold after 16 h of treatment with TNF-alpha and IL-6, respectively. Scatchard analysis of binding data showed that neither TNF-alpha nor IL-6 affected the affinity of IFN-gamma for its receptor. The increased surface expression of IFN-gamma R induced by TNF-alpha and IL-6 correlated with the rise in IFN-gamma mRNA receptor levels. TNF-alpha-mediated up-regulation of IFN-gamma R was due to an increase in transcriptional activity of the IFN-gamma R gene, as shown by run-on experiments. No significant modulation of IFN-gamma R gene transcription was observed in nuclei from cells treated with IL-6, whose effect appeared to be related to IFN-gamma mRNA receptor stabilization. Taken together, the present results provide, to our knowledge, the first evidence for cytokine-mediated up-regulation of IFN-gamma R in human monocytic cells. This up-regulation by TNF-alpha and IL-6, which are both produced by monocytes, occurs through different mechanisms, and may be of physiologic relevance in host defense.


Schistosoma mansoni eggs induce antigen-responsive CD44-hi T helper 2 cells and IL-4-secreting CD44-lo cells. Potential for T helper 2 subset differentiation is evident at the precursor level. Schistosoma mansoni eggs are potent inducers of biased Th2-like immune responses. Using a model system where mice are immunized with isolated schistosome eggs, we demonstrate that CD44 expression, up-regulation of which has been linked to Th cell development, is increased on Th2 cells. We also investigate the functional properties of CD44-lo Th cells recovered from the overtly Th2 environment constituted by lymph nodes draining sites of egg deposition. Production of high levels of IL-4, IL-5, and IL-10 by Th cells responding to egg Ag is shown to be the property of a subpopulation expressing CD44-hi. This population of Th cells cosegregates with a blasting subpopulation expressing more IL-4R (but similar amounts of IL-2R) than Th cells from normal mice. These results indicate that mature Th2 cells responding to schistosome eggs are CD44-hi and suggest that they use IL-4 as a growth factor. In contrast, CD44-lo cells sorted from lymph node populations responding to eggs are able to produce small amounts of IL-4 and IL-2, but no IL-5 or IL-10. This is surprising, because low expression of CD44 is considered a characteristic of Th cell naivite and concomitant ability to produce only IL-2. Thus, in lymph nodes responding to schistosome eggs, potential for Th2 subset differentiation is evident within the CD44-lo precursor Th subpopulation.


Soluble complement receptor type 1 ameliorates the local and remote organ injury after intestinal ischemia-reperfusion in the rat. We examined the role of C activation in ischemia reperfusion injury by inhibiting C activation in a rat model of mesenteric arterial occlusion. In anesthetized rats, 60 min of mesenteric arterial occlusion was followed by 3 h of reperfusion. PBS alone or containing soluble C receptor 1 (3 or 6 mg) was administered i.v. Controls underwent laparotomy without ischemia. Relative serum C activities were assessed by hemolytic assay, neutrophil (polymorphonuclear leukocyte) sequestration by tissue content of myeloperoxidase (MPO) activity, intestinal mucosal injury by histologic grading, lung vascular permeability by the ratio of bronchoalveolar lavage to blood concentration of radiolabeled BSA, and endothelial cell injury was quantified by measurement of plasma factor VIII-related Ag. After reperfusion, PBS-treated animals had increased intestinal MPO (0.048 +/- 0.007 U/g) compared to sham (0.022 +/- 0.005 U/g (p less than 0.05)) and intestinal mucosal injury score (2.490 +/- 0.221) compared to sham (0.331 +/- 0.045 (p less than 0.05)). Treatment with 6 mg soluble C receptor 1 15 min before reperfusion reduced intestinal MPO (0.017 +/- 0.003 U/g (p less than 0.05)) and mucosal injury (1.733 +/- 0.168 (p less than 0.05)) compared to PBS control. PBS-treated animals also demonstrated increased lung MPO (0.314 +/- 0.025 U/g vs 0.085 +/- 0.018 in sham (p less than 0.05)) and increased lung permeability (bronchoalveolar lavage/blood cpm 11.32 +/- 1.35 x 10(-3) vs sham 2.22 +/- 0.19 x 10(-3) (p less than 0.05)). Treatment with 6 mg soluble C receptor 1 15 min before reperfusion or at reperfusion reduced the lung permeability (bronchoalveolar lavage/blood cpm 3.90 +/- 0.79 x 10(-3) and 5.08 +/- 0.75, respectively (both p less than 0.05)) compared to PBS control, but did not reduce lung MPO (0.342 +/- 0.031 U/g and 0.246 +/- 0.025), respectively. Treatment with sCR1 also reduced the release of factor VIII-related Ag, 5-day mortality, and C hemolytic activity. In this model, C is a major mediator of intestinal injury and extraintestinal injury.


Protein tyrosine phosphorylation induced via the IgG receptors Fc gamma Ri and Fc gamma RII in the human monocytic cell line THP-1. We have investigated the role of protein tyrosine phosphorylation in transmembrane signaling via the IgG receptors Fc gamma RI and Fc gamma RII in the human monocytic cell line THP-1. Fc gamma RI and Fc gamma RII were selectively engaged using the anti-Fc gamma RI mAb 197 (IgG2a) and the anti-Fc gamma RII mAb IV.3 (IgG2b). Addition to cells of mAb 197, but not addition of IgG2a mAb of irrelevant specificity, resulted in the rapid induction of cytoplasmic protein tyrosine phosphorylation as assessed by antiphosphotyrosine immunoblotting. A similar pattern of tyrosine phosphorylation was induced by mAb IV.3, but not by control IgG2b mAb. The induction of tyrosine phosphorylation by anti-Fc gamma R mAb was not dependent on antibody Fc region-FcR interactions, because tyrosine phosphorylation was also induced by cross-linked anti-Fc gamma RI F(ab')2 fragments and by cross-linked anti-Fc gamma RII Fab fragments. To investigate the relationship of Fc gamma R-induced tyrosine phosphorylation and activation of phospholipase C, which is known to follow Fc gamma R engagement, we assessed the effect of the tyrosine kinase inhibitor herbimycin A on Fc gamma R-induced Ca2+ flux. Herbimycin A strongly inhibited cellular Ca2+ flux induced by mAb 197, but did not inhibit Ca2+ flux induced by aluminum fluoride, suggesting that tyrosine phosphorylation may be important in regulating Fc gamma R-mediated activation of phospholipase C. Consistent with this, mAb 197 induced rapid phosphorylation of the gamma-1 isoform of phospholipase C. Finally, herbimycin A strongly inhibited the induction of TNF-alpha mRNA accumulation by Fc gamma R cross-linking. These results suggest that protein tyrosine phosphorylation may play an important role in the activation of phospholipase C and in the induction of monokine gene expression that follows engagement of Fc gamma R in human monocytes.


Variability in T cell receptor V beta gene usage in human peripheral blood lymphocytes. Studies of identical twins, siblings, and insulin-dependent diabetes mellitus patients. Recent studies focused on the diversity and molecular organization of the human TCR-beta complex have begun to establish the genetic basis for the potential repertoire of V beta specificities in T cells. The scope and variability of the actual repertoire derived from this potential repertoire, however, remains to be clarified. In this study, V beta usage by human peripheral T cells derived from serial samples of the same individual, identical twins, and the members of three nuclear families that include four members with insulin-dependent diabetes mellitus (IDDM) was assessed by both quantitative polymerase chain reaction and Northern blotting with V beta subfamily-specific probes. Samples taken from the same individual over a period of 21 months and analyzed in separate experiments indicated stability in the peripheral repertoire, whereas the similarity in peripheral V beta usage in a pair of identical twins suggested a strong role for genetics in shaping the peripheral T cell repertoire. In contrast, V beta usage in siblings and in unrelated individuals was observed to differ substantially. In particular, peripheral expression of V beta 3 and V beta 20 differed by more than sixfold among members of two different families. Segregation analysis of TCR and HLA haplotypes in these families suggested that variation in V beta 20 expression was TCR haplotype specific. Subsequent nucleotide sequence analysis of the V beta 20 gene segment in multiple members of these families revealed the presence of a null allele for V beta 20 expression. No consistent significant differences in V beta usage were observed in IDDM patients relative to their siblings or between identical twins discordant for IDDM. These results suggest that the repertoire of peripheral T cell specificities present in different individuals in human populations varies dramatically because of the effects of multiple factors, including TCR germ-line polymorphism.


[Reconstruction for malignant bone tumor defects by a composite graft--cementless arthroplasty]. Five patients with malignant bone tumor were arthroplastied with Dacron fabric-enveloped prosthesis without using cement after resection of tumor. Usually following the wide resection of the primary lesion, the supporting tissues are always lost. Dacron fabric is intended to act as a scaffold upon which connective tissue can proliferate and form new ligament and supporting soft tissues. Dacron fabric is useful for the repair of ligament, muscles, joint capsule, periosteum, and other supporting soft tissues without significant complications. The lengths of the released supporting tissues are easily adjustable. Biologic fixation of the prosthesis to the soft supporting tissues is successful, then the durability of the prosthesis prolongs. The Dacron fabric is capable of inducing and supporting the ingrowth of vascular connective tissue which becomes stronger thereafter. The author believes that this method provides a stable, functional and esthetically pleasing reconstruction.


Muscular changes in osteoarthritis of the hip and knee. Muscular abnormalities in the patients with degenerative osteoarthritis were examined histologically and histochemically. Enzyme stainings for DPNH-tetrazolium reductase and myosin ATP-ase at pH 10.4 and 4.6 were performed on 26 vastus lateralis biopsies from the patients with osteoarthritis of the knee and 21 gluteus medius specimens from those with osteoarthritis of the hip. All specimens were obtained at surgery. The patients were all female with their ages ranging from 40 to 74 years. Control specimens were obtained from 16 fractured females with absence of arthritis. Type 2 fiber atrophy and internal derangement of fibers were found in both osteoarthritic and control muscles, but were more frequently seen in the vastus lateralis of the patients with osteoarthritis of the knee. Abnormal mosaic patterns of fiber types, such as fiber type grouping and grouped atrophy of type 2 fibers, were frequently found in the vastus lateralis of the patients with osteoarthritis of the knee (73.1%), and less frequently in that of controls (6.3%). However, abnormal mosaic patterns were not found in the gluteus medius obtained from either osteoarthritic patients or control subjects. Abnormal changes in mosaic pattern of fiber types in the vastus lateralis of osteoarthritis of the knee may suggest associated motor neuron dysfunction; the abnormality was not observed in the gluteus medius specimens of the patients with osteoarthritis of the hip, for which a different background in neurological factors may be involved.


Comparison of the effect of a proteolytic enzyme dispase and a chelator for the preparation of epidermal sheet from nude mice. Histochemical studies of epidermal Langerhans cells require well-preserved epidermal sheets. We studied the conditions for the preparation of the epidermal sheets from the skin of the ear, hind limb and trunk of nude mice. Two types of commercial dispase, and ethylenediaminetetraacetic acid (EDTA), were used at several concentrations and the effects were compared. Dermo-epidermal separation was evaluated by the preservation of ATPase activity of Langerhans cells in the epidermal sheet and by the ultrastructure of epidermal cells (Langerhans cells and keratinocytes) as well as epidermal-dermal junction. Good separation depended on the combination of the concentration of the reagent and site of the skin. The concentrations of both dispase and EDTA effective for separation of the epidermis in the nude mouse were lower than those in other mouse strains reported.


A death involving probenecid.

A death involving probenecid. A death following deliberate ingestion of approximately 75 g of probenecid in a 36-year-old man is described. Tissue concentrations of probenecid were highest in serum (710 mg/L) and liver (550 mg/kg). Probenecid was also detected in vitreous and bile. Ethanol was also detected in blood at 0.13 g/100 mL.


Work-related injuries in Athens County 1982 to 1986. A comparison of emergency department and workers' compensation data. In 1987, the Panel on Occupational Safety and Health Statistics issued a report concluding that the existing national surveillance system for occupation injuries might result in substantial underreporting of occupational injuries. In this study, we examined two sources of data on occupational injuries, the National Electronic Injury Surveillance System (NEISS) and lost-work time claims to the Bureau of Workers' Compensation (BWC), available in one community, Athens County, Ohio. Based on comparison of the NEISS and BWC data sets, we conclude that neither data set alone gives a complete nor an accurate picture of occupational injuries in Athens County. The two may provide a more complete representation of occupational injuries when examined together. Using the NEISS and BWC data sets in combination results in a total number of injuries higher than that predicted by national norms.


Young children with disabilities and assistive technology: the nurse's role on multidisciplinary technology teams. Considerations for the involvement of nurses in the provision of assistive technology in education and intervention settings as members of multidisciplinary teams are presented. Philosophical considerations when participating in these processes are discussed, followed by a delineation of factors important in the selection of appropriate technology. Recommendations are discussed that have practical implications for nursing professionals.


Brainstem 5-hydroxytrytamine1A binding sites are not down-regulated by agonists which induce tolerance in the rat: myoclonus and other serotonergic behaviors. To study the regulation of 5-HT1A receptors in the brainstem, the region most relevant to the serotonin syndrome and to serotonin-responsive human myoclonic disorders, we chronically treated rats with various 5-HT1A agonists and labeled 5-HT1A sites with [3H]8-OH-DPAT. Daily injection for 30 consecutive days of 10 mg/kg ip 8-OH-DPAT (pre- and post-synaptic 5-HT1A agonist) significantly decreased 8-OH-DPAT-evoked flat body posture, forelimb myoclonus, and hypothermia compared to chronic vehicle injection. There was no cross tolerance to 8-OH-DPAT in rats chronically injected with ipsapirone or buspirone (presynaptic 5-HT1A agonists). However, none of the 5HT1A agonists significantly altered Bmax of brainstem 5-HT1A binding sites. Chronic injection with other drugs such as 1-propranolol, (+/-) pindolol and spiperone (5-HT1A and 5-HT2 antagonists), methysergide (5-HT1 and 5-HT2 antagonist), and agonists and antagonists at various other 5-HT receptors also had no effect on binding parameters. These data demonstrate lack of cross-tolerance between pre- and post-synaptically acting 5-HT1A agonists and absence of down-regulation of presynaptic 5-HT1A sites at doses which induced tolerance of 5-HT1A-mediated behaviors of the serotonin syndrome. They suggest changes in the post-synaptic cell rather than the receptor recognition site as the mechanism of tolerance.


A low affinity vasopressin V2-receptor in inherited nephrogenic diabetes insipidus. Congenital nephrogenic diabetes insipidus (NDI) is an X-linked inherited disorder characterized by renal resistance to the antidiuretic hormonal action of vasopressin. This study describes the molecular basis of nephrogenic diabetes insipidus in a dog family. Kidney membranes prepared from NDI-affected male huskies were examined for vasopressin binding and response. Compared to membranes from unaffected canines, those from the kidney inner medulla of NDI-dogs possessed normal V2-receptor numbers, but with 10-fold lower affinity for [Arg8] vasopressin (AVP). Adenylate cyclase stimulation by AVP in contrast to that by forskolin or GTP-analogues was similarly reduced in a dose responsive manner. The NDI-affected dogs showed antidiuretic responses to very high doses of V2-specific agonists, consistent with their possessing V2-receptors of lower affinity. Prolonged treatment with V2-agonists, 1-deamino [D-Arg8] VP (dDAVP) and 1-deamino [Val4, Sar7] AVP (dVSAVP), rendered the NDI-affected dogs near normal in terms of water intake and urine osmolality.


Delayed implantation. A case report.

Delayed implantation. A case report. In cycles in which conception takes place, serum human chorionic gonadotropin (hCG) becomes detectable between 8 and 12 days after ovulation. A delayed appearance of hCG has been reported in a limited number of cases, most of them ending in spontaneous abortion. We encountered a case of ectopic pregnancy characterized by a delayed appearance of hCG and accompanied by a complete, albeit temporary, halt in the steroidogenic activity of the corpus luteum. Although the patient was at risk of developing an ectopic pregnancy, the findings made an early diagnosis extremely difficult.


NMDA antagonist activity of (+/-)-(2SR,4RS)-4-(1H-tetrazol-5-ylmethyl)piperidine-2-carboxylic acid resides with the (-)-2R,4S-isomer. The tetrazole-substituted amino acid (+/-)-(2SR,4RS)-4-(1H-tetrazol-5-ylmethyl)pip eri dine-2-carboxylic acid (LY233053, (+/-)-1) was resolved into its constituent enantiomers by treatment of a key intermediate in the synthesis of the racemic amino acid, ethyl (+/-)-cis-4-(cyanomethyl)-N-allylpiperidine-2-carboxylate, with either 2S,3S- or 2R,3R-di-p-toluoyltartaric acid. These resolved amines were then converted as for the racemate to the amino acids (-)-1 and (+)-1. The activity of this potent and selective NMDA antagonist was found to reside with the (-)-isomer of 1 (LY235723). X-ray crystallographic analysis of the 2S,3S-di-p-toluoyltartaric acid salt of ethyl cis-4-(cyanomethyl)-N-allylpiperidine-2-carboxylate showed that the resolved amine, and thus (-)-1, possessed the 2R,4S absolute stereochemistry. Affinity for the NMDA receptor was determined using the specific radioligand [3H]-(2SR,4RS)-4-(phosphonomethyl)piperidine-2-carboxylic acid ([3H]CGS 19755; IC50 = 67 +/- 6 nM), and selective NMDA antagonist activity was determined using a cortical slice preparation (IC50 versus 40 microM NMDA = 1.9 +/- 0.24 microM). This compound also demonstrated potent NMDA antagonist activity in vivo following systemic administration through its ability to block NMDA-induced convulsions in neonatal rats, NMDA-induced lethality in mice, and NMDA-induced striatal neuronal degeneration in rats.


Occurrence of mycotoxins in animal feeds. A total of 1120 grain samples of oats, wheat, rye, barley, and maize, delivered for processing of mixed feeds for animals, were collected during the years 1975 to 1979 from commercial feed mills located throughout Poland. In addition, 625 samples of the commercially mixed feeds and protein concentrates were collected during 1976. For the mycotoxin survey, 751 laboratory samples were chosen at random and analyzed. When applying confirmatory tests neither aflatoxins B1, B2, G1, G2 nor sterigmatocystin, zearalenone, or ochratoxin B were found to be present in any of the samples of barley, wheat, rye, or oats. Aflatoxins were detected in about 4% of the maize samples. The presence of ochratoxin A in the range of 2 to 200 micrograms/kg was evident in 9% of the grain samples. The commercially mixed feeds were found to be more contaminated with mycotoxins than were the grains. The aflatoxins were confirmed in about 13% of the samples of mixed feeds. After the preliminary multimycotoxin analysis, out of 42 feed samples that could be suspected of containing ochratoxins, 32 failed to prove their presence. Similarly, out of 27 suspected feed samples, zearalenone was found only in one (0.5%) sample. The lowest percentage of samples contaminated with mycotoxins was found in poultry mixed feeds (4%). The highest contamination occurred among the samples of swine rations, where 17% of the samples contained aflatoxins and 13% ochratoxins. The protein concentrates contained only aflatoxins. Out of 31 analyzed samples, aflatoxins were detected in 19 (61%) in concentrations ranging from 5 micrograms/kg to 500 micrograms/kg. In one sample, aflatoxin concentration (B1 + B2) reached 1140 micrograms/kg. Practical implications of the results are discussed in relation to animal and human safety.


Dihydropyrimidine calcium channel blockers. 4. Basic 3-substituted-4-aryl-1,4-dihydropyrimidine-5-carboxylic acid esters. Potent antihypertensive agents. We have examined a series of novel dihydropyrimidine calcium channel blockers that contain a basic group attached to either C5 or N3 of the heterocyclic ring. Structure-activity studies show that a 1-(phenylmethyl)-4-piperidinyl carbamate moiety at N3 and sulfur at C2 are optimal for vasorelaxant activity in vitro and impart potent and long-acting antihypertensive activity in vivo. One of these compounds (11) was identified as a lead, and the individual enantiomers 12a (R) and 12b (S) were synthesized. Two key steps of the synthesis were (1) the efficient separation of the diastereomeric ureido derivatives 29a/29b and (2) the high-yield transformation of 2-methoxy intermediates 30a/30b to the (p-methoxybenzyl)thio intermediates 31a/31b. Chirality was demonstrated to be a significant determinant of biological activity, with the dihydropyridine receptor recognizing the enamino ester moiety (12a) but not the carbamate moiety (12b). Dihydropyrimidine 12a is equipotent to nifedipine and amlodipine in vitro. In the spontaneously hypertensive rat, dihydropyrimidine 12a is both more potent and longer acting than nifedipine and compares most favorably with the long-acting dihydropyridine derivative amlodipine. Dihydropyrimidine 12a has the potential advantage of being a single enantiomer.


Stability of coliphage lambda DNA replication initiator, the lambda O protein. The initiator of coliphage lambda DNA replication, lambda O protein, may be detected among other 35S-labeled phage and bacterial proteins by a method based on immunoprecipitation. This method makes it possible to study lambda O proteolytic degradation in lambda plasmid-harboring or lambda phage-infected cells; it avoids ultraviolet (u.v.)-irradiation of bacteria, used for depression of host protein synthesis, prior to lambda phage infection. We confirm the rapid decay of lambda O protein (half-time of 80 s), but we demonstrate the existence of a stable lambda O fraction. In the standard five minute pulse-chase experiments, 20% of synthesized lambda O is stable. The extension of the [35S]methionine pulse, possible in lambda plasmid-harboring cells, leads to a linear increase of this fraction, as if a part of the synthesized lambda O was constantly made resistant to proteolysis. Less than 5% of lambda O protein synthesized during one minute is transformed into a stable form. We presume that the stable lambda O is identical with lambda O present in the normal replication complex and thus protected from proteases. We cannot find any stable lambda O in Escherichia coli recA+ cells that were irradiated with u.v. light prior to lambda phage infection, but their recA- counterparts behave normally, suggesting that recA function interferes in the assembly of a normal replication complex in u.v.-irradiated bacteria. The stable lambda O found in lambda plasmid-harboring, amino acid-starved relA cells is responsible for the lambda O-dependent lambda plasmid replication that occurs in this system in the absence of lambda O synthesis. The existence of stable lambda O raises doubt concerning its role as the limiting initiator protein in the control of replication. Another significance of lambda O rapid degradation is proposed.


Inheritance of the replication complex by one of two daughter copies during lambda plasmid replication in Escherichia coli. Direct measurement of DNA synthesis confirmed that lambda plasmid replication proceeds for several hours in an amino acid-starved relA mutant of Escherichia coli, leading to plasmid amplification; this replication is lambda cro-independent, but requires the function of lambda O initiator in the absence of its synthesis. This suggests that after the assembly of the replication complex (RC) at ori lambda the lambda O protein remains in this structure and the affinity of lambda O to ori lambda is alleviated in the assembled RC allowing its movement along the DNA. During amino acid starvation the lambda plasmid DNA synthesis per bacterial mass occurs at a constant level, as would be expected if the number of functioning RCs remained constant. This favors the idea that under these conditions the next replication round operates due to the activity of the RC inherited from the preceding round. Density shift experiments reveal indeed that, from two daughter plasmid copies synthesized after the onset of amino acid starvation only one is able to enter into the next round of replication. We infer that this is the plasmid copy that inherits the lambda O-enclosing RC from the previous replication round. Moreover, the same results of density shift experiments were obtained for plasmids synthesized before the onset of amino acid starvation. Therefore, we presume that in lambda plasmid-harboring bacteria growing in nutrient medium, every second plasmid circle bears an RC that originates from the preceding round of replication. This structure has to be assembled de novo only on the daughter plasmid copy that does not inherit the parental RC. In the absence of lambda O initiator synthesis in amino acid-starved relA cells this process cannot occur, leaving as the only replication pathway that driven by the parental RC. Our results are discussed in relation to the model of regulation of lambda plasmid replication.


Isolated systolic hypertension and subclinical cardiovascular disease in the elderly. Initial findings from the Cardiovascular Health Study. To assess the association between isolated systolic hypertension (ISH) and subclinical disease in adults aged 65 years and above. Medicare eligibility lists were used to obtain a representative sample of 5201 community-dwelling elderly persons for the Cardiovascular Health Study, a National Heart, Lung, and Blood Institute--sponsored cohort study of risk factors for coronary heart disease and stroke. In this cross-sectional analysis of baseline data, we excluded 3012 participants who were receiving antihypertensive medications, had clinical cardiovascular disease, or had a diastolic blood pressure of at least 90 mm Hg. For electrocardiogram: myocardial infarction, left ventricular hypertrophy, and left ventricular mass as measures of myocardial damage and strain; for echocardiography: left ventricular mass, fractional shortening, and Doppler flow velocities as measures of cardiac systolic and diastolic function; and for carotid sonography: carotid arterial intima-media thickness as a measure of atherosclerosis. Among the 2189 men and women in this analysis, 195 (9%) had ISH (systolic blood pressure, greater than or equal to 160 mm Hg) and 596 (23%) had borderline ISH (systolic blood pressure, 140 to 159 mm Hg). Systolic blood pressure was associated with myocardial infarction by electrocardiogram (P = .02). Borderline and definite ISH were strongly associated with left ventricular mass (P less than .001). While there was little association with cardiac systolic function, borderline and definite ISH were associated with cardiac diastolic function (P less than .001). Isolated systolic hypertension was also strongly associated with increased intima-media thickness of the carotid artery (P less than .001). While cohort analyses of future repeated measures will provide a better assessment of risk, both borderline and definite ISH were strongly related to a variety of measures of subclinical disease in elderly men and women.