The beneficial effects of a thromboxane receptor antagonist on spinal cord perfusion following experimental cord injury.

The eicosanoids thromboxane A2 and prostacyclin have opposing actions causing vasoconstriction and vasodilation respectively. The ratio of these two eicosanoids is thus an important determinant of circulatory homeostasis. An increase in this ratio occurs in certain inflammatory conditions with dramatic consequences in organ perfusion. In spinal cord trauma, in addition to direct physical perturbation of the spinal cord, it is likely that further structural and functional loss occurs as a result of decreased tissue perfusion precipitated by an increase in the thromboxane/prostacyclin ratio. This study evaluated hemodynamics and organ perfusion, 3 h following 24 g-cm spinal cord trauma in the rat. The role of thromboxane was investigated with an inhibitor of thromboxane synthesis (Dazoxiben) and with a receptor antagonist (13-APT). Cardiac output and blood pressure were unaffected by Dazoxiben, 13-APT, or spinal cord trauma. Injury effected approximately a 40% decrease in spinal cord perfusion from 0.41 to 0.25 ml/min/g which was not improved by the thromboxane synthase inhibitor, Dazoxiben. 13-ATP completely abrogated the decline in spinal cord blood flow flowing injury. Perfusion of other selected organs demonstrated little change as a result of the spinal trauma. Brain flow remained constant at 0.78 ml/min/g brain. Coronary blood flow, however, declined from 3.2 to 2.0 ml/min/g heart tissue. The data suggest consideration of the importance of thromboxane in therapeutic attempts to reduce secondary injury arising in spinal cord trauma.


In-vivo SPECT imaging of D2 receptor with iodine-iodolisuride: results in supranuclear palsy.

We assessed the potential use of [123I]iodolisuride (ILIS), a new iodine ergolene derivative, to study human striatal D2 dopamine receptors with SPECT. In normal subjects, we found that the tracer accumulated preferentially in striatum. This was prevented by high doses of haloperidol. The striatal accumulation was maximal between 60 and 180 min after injection. The striatum-to-cerebellum radioactivity concentration ratio as an index of specific binding, measured 60 min after injection, was 1.52 +/- 0.19 (mean +/- s.d.) in controls and 1.36 +/- 0.11 in patients with supranuclear palsy (p less than 0.03). Our results show that ILIS may be used to study D2 receptors with SPECT. In-vivo changes of D2 receptors in human brain may be detected with this method.


Angiotensin converting enzyme inhibitors and the cardiovascular system.

To show that angiotensin converting enzyme (ACE) inhibitors benefit the cardiovascular system to a greater extent than other antihypertensive agents. An extensive literature search has shown that ACE inhibitors can induce regression of left ventricular hypertrophy, which may improve systolic and/or diastolic function in the left ventricle; increase in arterial compliance; and improve clinical signs of congestive heart failure and reduce mortality in patients with severe heart failure. In addition, ACE inhibitors have either a neutral or a beneficial effect on blood lipids and lipoproteins. These agents also have potential anti-ischaemic and anti-arrhythmic activity but do not depress resting cardiac function. ACE inhibitors are uniquely valuable first-line antihypertensive agents, especially in patients with cardiovascular risk.


Immunohistochemical studies on the innervation of human transplanted liver.

The changing pattern of innervation in the human transplanted liver was studied from the day of transplantation to 5 years later. Seven liver biopsies from non-transplant controls, 37 liver biopsies from 22 transplant patients, and one of these biopsied livers removed at retransplantation, were available for the study. Sections were immunostained for protein gene product 9.5 (PGP 9.5), neurone-specific enolase (NSE), S-100 protein, and vasoactive intestinal polypeptide. NSE and PGP 9.5 demonstrated nerves most successfully in our tissues. Staining for most small nerves was reduced by day 5 post-transplantation. Scanty fine nerves could be detected from day 13 to day 241 in occasional biopsies. Consistently identifiable immunostaining of PGP 9.5 and NSE nerve fibres was again apparent in portal areas after this time in all but one case. The findings indicate that in transplanted liver limited reinnervation can eventually take place. This could be due to either proliferation of intrinsic nerves, or regrowth of extrinsic nerves, or both.


Hepatic reinnervation following orthotopic liver transplantation in man.

We have studied changes in the pattern of intrinsic hepatic innervation in sequential liver biopsies from 16 patients who underwent orthotopic liver transplantation. Seventy-one needle biopsies were used, including specimens obtained at the time of transplantation (time zero) and up to 4 years post-transplantation; five transplant hepatectomy tissue blocks removed 3-32 months after transplantation were also assessed. Paraffin sections were immunostained with anti-PGP 9.5 and anti-S-100 to identify nerve fibres. All 'time zero' biopsies contained portal nerves and all but two showed staining of parenchymal fibres. After 1 week, no subsequent biopsies contained parenchymal fibres. The disappearance of portal fibres was less rapid and showed greater variability between patients, but they had all disappeared by 6 weeks and there was no positive staining between 6 and 60 weeks. Thereafter, a minority of biopsies showed innervation of a few small portal tracts. Samples from the porta hepatis, hepatectomy specimens, and needle biopsies containing large tracts showed persistence of major nerve trunks at all stages. Abnormally large nerve bundles were seen in some of these areas. The pattern of nerve staining showed no obvious relationship to the intensity of rejection changes. Our results suggest that there is a limited, delayed capacity for regeneration of portal, but not parenchymal, fibres in the transplanted human liver. The physiological significance of this long-term parenchymal denervation in transplanted livers remains to be determined.


Isoenzyme analysis of Hammondia hammondi and Toxoplasma gondii sporozoites.

Isoenzyme analysis using isoelectrofocusing in polyacrylamide gels was used to distinguish Hammondia hammondi and Toxoplasma gondii sporozoites. Five enzyme systems were studied: aconitase (EC 4.2.1.3), aspartate aminotransferase (EC 2.6.1.1), glucose phosphate isomerase (EC 5.3.1.9), lactate dehydrogenase (EC 1.1.1.27), and phosphoglucomutase (EC 2.7.5.1). Three stocks of T. gondii belonging to 3 zymodemes were compared to 1 stock of H. hammondi. Hammondia hammondi differed from T. gondii at all 5 loci analyzed. This was observed for all 3 zymodemes of T. gondii. These results indicated clear genetic differences between the 2 species.


Comparison of isozyme patterns between Spirometra erinacei and Spirometra mansonoides by isoelectric focusing.

No differences were observed in the isozyme patterns of 4 enzymes examined between fresh samples stored at -80 C and samples stored at room temperature for 10 days after lyophilization, which supports the validity of comparing lyophilized samples to fresh frozen tissue. Mature proglottids as well as plerocercoids of Spirometra erinacei from Japan and Australia were indistinguishable by comparison of isozyme patterns after isoelectric focusing. The isozyme patterns of acid phosphatase, glucosephosphate isomerase (GPI), and mannosephosphate isomerase from plerocercoids of Spirometra mansonoides were distinctly different from those of plerocercoids of S. erinacei. The adenylate kinase isozyme patterns of the mature proglottids of S. mansonoides were also distinctly different from those of the mature proglottids and the plerocercoids of S. erinacei. The GPI isozyme pattern of the mature proglottids of S. mansonoides was also distinguishable from the GPI patterns of those of S. erinacei. These electrophoretic data suggest that the S. erinacei from Japan and Australia are closely related, if not identical, but that S. mansonoides is genetically distinct from S. erinacei.


CD8+ T lymphocytes are not required for murine resistance to human filarial parasites.

Mice are resistant to the establishment of infection with the nematode parasite Brugia malayi, an etiologic agent of human lymphatic filariasis. We have recently shown that T and B lymphocyte-deficient C.B.-17 scid/scid mice are permissive for infection with this parasite, whereas coisogenic C.B.-17+/+ mice are resistant. This observation suggests that T and B lymphocytes that comprise the antigen-specific immune system orchestrate murine resistance to B. malayi. In order to define the component of the antigen-specific immune response that is responsible for this resistance, we have tested the susceptibility of beta 2M-/- mice to infection with B. malayi L3 larvae. These mice are homozygous for insertional disruption of their B2m genes, which encode beta 2-microglobulin, the small subunit of the major histocompatibility (MHC) antigens. They do not express beta 2-microglobulin and, as a consequence, fail to express the class I major histocompatibility antigens, and they do not develop the CD8+ class I MHC-restricted cytotoxic T cell subset. We find that these mice are completely resistant to B. malayi, indicating that the CD8+ T lymphocyte subset is not an obligate requirement for murine resistance to human filarial parasites.


Preferred antagonist binding state of the NMDA receptor: synthesis, pharmacology, and computer modeling of (phosphonomethyl)phenylalanine derivatives.

A series of substituted [phosphono-, sulfo-, carboxy-, and (N-hydroxycarbamoyl)methyl]phenylalanines were synthesized as probes for the investigation of the preferred antagonist state of the NMDA receptor antagonists. The potency of these compounds was evaluated by measuring electrophysiological responses induced by NMDA in cultured mouse cortical neurons. 3-(Phosphonomethyl)phenylalanine [1(m)] a formal AP7 analogue, has been shown to be the most potent antagonist in this study with an IC50 of around 5 microM. The isomeric 2-(phosphonomethyl)phenylalanine [1(o)] was about half as active as 1(m) and as active as compound 5(3), a derivative which is cis-hydrogenated on the phenyl ring of 1(m). Replacement of a phosphono by a sulfo group led to a large reduction in the ability of these compounds to antagonize NMDA responses, although the ortho and meta isomers retained some activity in their reduced forms. In both series the para isomers were almost completely inactive at 100 microM. Introduction of a carboxyl or a bidentate HONHCO group in place of the phosphono moiety in the 3-position results in compounds devoid of activity. The active and inactive compounds of this study were used in conjunction with the most potent linear and cyclic phosphono-containing NMDA antagonists reported to date to determine, via computer modeling techniques, a three-dimensional model corresponding to a antagonist preferring state of the NMDA binding site. This structure defines a pharmacophore which is characterized by (i) well-defined distances between the central atoms of the polar groups PO3H-, NHn+, (n = 2, 3), and COO- (P-N = 5.89 +/- 0.12 A, P-C = 6.66 +/- 0.08 A, and N-C = 2.28 +/- 0.01 A), (ii) a sterically allowed region between the C5 methylene and the PO3H- group, and (iii) a molecular electrostatic field in which the positive, neutral, and negative potential zones are self-contained--with the negative potential zone connecting the PO3H- and COO- groups as the largest. We have compared our results to a preliminary model of the NMDA antagonist site by Hutchison et al. and to a topological model of the NMDA-glycine receptor site by Cordi et al. Our proposed steric-electrostatic pharmacophore which refines, simplifies, and improves these models has now to be validated by the design of new NMDA antagonists.


Nuclear DNA content and nucleation patterns in rat cardiac myocytes from different models of cardiac hypertrophy.

Nuclear DNA content and number of nuclei were examined in cardiac myocytes isolated from controls and rats with volume and pressure overload hypertrophy to determine if haemodynamic overload alters these nuclear parameters. The experimental groups were comprised of normotensive (WKY) and Spontaneously Hypertensive rats (SHR). Additionally, Sprague-Dawley rats with aortic constriction (AC), pulmonary stenosis (PS), myocardial infarction (MI), and 5 month arteriovenous fistulas (F) were studied along with appropriate shams for each of these groups. Nuclear DNA content was measured from DAPI-stained nuclei using an image analysis microdensitometry system. Myocyte volume was measured with a Coulter Channelyzer system. Approximately 83% of the left ventricular myocytes from the SHR and WKY groups contained a diploid DNA content with the remainder being tetraploid. The remaining experimental and sham groups, all female Sprague-Dawley rats (SD), were approximately 93% diploid. The nucleation patterns differed slightly between rat strains with the SHR/WKY expressing approximately 85% binucleation, 14% mononucleation and 5% tri- or tetranucleation. All SD groups, control and hypertrophied, showed approximately 89% binucleation, and 10% mononucleation with the remainder being tri- or tetranucleated. In summary: (1) cardiac myocytes from SHR/WKY strains are predominantly diploid but to a lesser degree than myocytes from SD; (2) nuclear number follows the same pattern with SHR/WKY showing a smaller percentage of binucleated myocytes than SD myocytes; (3) neither the duration, severity, or type of overload caused a significant change in the extent of polyploidy in overloaded hearts from SD rats; and (4) the extent of polyploidy in cardiac myocytes from both the right and left ventricles of SHR and WKY animals does not differ statistically.


Inverse modulation of steady-state messenger RNA levels of two non-integrin laminin-binding proteins in human colon carcinoma.

Interactions between cells and the basement membrane glycoprotein laminin are altered during colon cancer progression. Colon carcinoma and normal mucosa cells express a variety of laminin-binding proteins, including the 67-kd laminin receptor (67 LR) and a 31-kd human laminin-binding protein (HLBP31) homologous to the 31-kd human IgE-binding protein/galactoside-binding lectin. To investigate whether various laminin-binding proteins are differentially expressed in human colon carcinoma, we studied messenger RNA (mRNA) levels of the 67 LR and HLBP31 in matched tumor and adjacent normal mucosa samples from a series of 21 patients. Total cellular RNA from tumor and normal mucosa was isolated and analyzed by Northern and slot blot hybridization. In addition, HLBP31 protein levels were assessed by the immunoblot technique. Quantitative laminin affinity chromatography was also used to measure the synthesis of HLBP31 protein in five human cancer cell lines. The steady-state mRNA level of HLBP31 was downregulated (i.e., decreased) in 18 of 21 human colon carcinomas compared with the level in their corresponding normal colonic mucosa. On average, the level of HLBP31 mRNA was decreased 50% +/- 30% (+/- SD) in the colon cancers. The mean ratio of colon cancer HLBP31 mRNA to adjacent normal mucosa HLBP31 mRNA was twofold lower in primary tumors of patients with metastases (0.3 +/- 0.2 SD) than in primary tumors of patients free of metastatic lesions (0.6 +/- 0.2 SD). The differences between the two groups of patients were statistically significant (P less than .05, Wilcoxon-Mann-Whitney test). We have previously shown that the ratio of colon cancer 67 LR mRNA to corresponding normal mucosa 67 LR mRNA was increased in the same patient population. When the two ratios (ratio of cancer to normal HLBP31 mRNA and ratio of cancer to normal 67 LR mRNA) were compared, HLBP31 mRNA/67 LR mRNA was significantly lower (P less than .05) in primary tumors with metastases (mean +/- SD, 0.3 +/- 0.2) than in primary cancers without metastases (mean +/- SD, 0.7 +/- 0.5). The steady-state level of HLBP31 mRNA was directly correlated with the amount of HLBP31 protein in both colon tissue samples and human cancer cell lines. HLBP31 mRNA expression in colon cancer tissues is modulated inversely to that of 67 LR mRNA expression. The down-regulation of HLBP31 appears to be associated with the metastatic capabilities of colon cancer cells. Prospective studies on a large cohort should determine if the systematic detection of HLBP31 and 67 LR protein and/or mRNA can be a valuable adjunct in the prognostic evaluation of primary colon cancers.


Prophylactic antibiotics for the prevention of infectious complications including empyema following tube thoracostomy for trauma: results of meta-analysis.

Since 1977, six clinical trials have been performed on the subject of routine antibiotic prophylaxis in patients requiring tube thoracostomy for trauma. No definitive conclusions have been reached regarding the efficacy of antibiotic use in this setting. The results of these clinical trials were pooled to generate an unbiased estimate of the efficacy of antibiotic prophylaxis for tube thoracostomy using the technique of meta-analysis. Meta-analysis is a statistical method for synthesizing results from separate but similar experiments, grouping them, and comparing each to the null hypothesis. Meta-analysis allows synthesis of all of the available data on antibiotic prophylaxis for tube thoracostomy to resolve the controversy surrounding this issue generated by different but similar clinical studies with conflicting results. Despite different conclusions of value when taken individually, the combined analysis does not support the null hypothesis (no effect of antibiotics). The statistical method is highly significant despite different mechanisms of injury, pathologic findings, and antibiotics employed.


The alterations of norepinephrine and acetylcholine concentrations in immature rat urinary bladder caused by streptozotocin-induced diabetes.

The concentrations of norepinephrine (NE) and acetylcholine (ACh) in the body and base of the bladder were investigated two, four and eight weeks after the administration of streptozotocin to four-week-old rats. At four and eight weeks after following dosage, NE concentrations in the base of the bladder of the diabetic rats were significantly higher than those of control rats. At the two-week diabetic condition, ACh concentrations in both the body and base of the bladder diabetic rats were about two-fold higher than in the respective control levels and remained constant for eight weeks. However, the control rats showed dramatic increase in ACh concentration from two to four weeks after dosage. There was no significant difference in the response of bladder muscle strips to phenylephrine, isoproterenol and ACh in the diabetic vs. control rats throughout the experiment. These observations suggest that the alteration of autonomic nervous system may occur at an early stage of diabetes mellitus in immature rats and that the high concentration of NE in the diabetic bladder base may reflect an adaptive overfunction of the adrenergic system probably to maintain continence. Furthermore, the lack of change in ACh concentration with age in diabetic rats may suggest an impaired development of the cholinergic system.


IL-1 receptor antagonist inhibits monocyte chemotactic peptide 1 generation by human mesangial cells.

The elicitation of neutrophils and monocytes from the circulation into the inflamed glomerulus is a key process in the pathogenesis of proliferative glomerulonephritis. The aim of this study was to determine the factors which regulate the expression and synthesis of the monocyte specific chemotaxin, monocyte chemotactic peptide 1 (MCP-1). Mesangial cells in culture did not constitutively express MCP-1, but could be induced to express both MCP-1 mRNA and antigenic MCP-1 by either stimulation with IL-1 alpha or TNF alpha, which are also stimuli for interleukin 8 (IL-8/NAP-1) expression and release. Pre-treatment of mesangial cells with the IL-1 receptor antagonist (IL-1ra) induced dose-dependent inhibition of both the expression of MCP-1 and IL-8 mRNA as well as the release of both chemotactic peptides in response to IL-1 alpha, while the receptor antagonist had no significant effect on TNF alpha induced MCP-1 and IL-8 generation. This study demonstrates that the IL-1 receptor antagonist was four times more effective at inhibiting the IL-1 induced expression and release of IL-8 compared to that of MCP-1. These results suggest that mesangial cell-derived MCP-1 may play an important role in the recruitment of monocytes in glomerular inflammation and that an IL-1 receptor antagonist may have therapeutic potential for the treatment of glomerulonephritis.


Improved heart failure protection by FK664, a novel positive inotropic agent, in dog heart-lung preparations.

The effects of FK664, a novel positive inotropic agent, and enoximone on pentobarbital-induced heart failure were compared in dog heart-lung preparations. Both FK664 and enoximone improved the cardiac function curve in a dose-dependent manner and restored it to the control level at drug concentrations of 1 microgram/ml and 10 micrograms/ml, respectively. Therefore, the cardiotonic potency of FK664 appears to be 10 times that of enoximone. These agents were almost equal in force-rate separation of cardiac effect. Neither of the agents produced arrhythmia at any dose tested. These results suggest that FK664 may be a potent cardiotonic agent for the treatment of heart failure.


Characterization of binding of a specific antagonist, [3H]-SQ 29,548, to soluble thromboxane A2/prostaglandin H2 (TP) receptors in human platelet membranes.

Binding of [3H]-SQ 29,548 was characterized to soluble thromboxane A2/prostaglandin H2 (TP) receptors from human platelet membranes as a means of examining ligand-receptor interactions outside the lipophilic environment of the cell membrane. Kinetic determination revealed a rate of ligand-receptor association of 1.4 x 10(7) +/- 0.2 M-1 x min-1 and a rate of dissociation of 0.5 +/- 0.07 min-1. The resultant equilibrium affinity constant was 36.3 +/- 5.8 nM. Saturation binding analysis revealed a single class of [3H]-SQ 29,548 binding sites with an affinity constant of 39.7 +/- 4.3 nM and a B(max) of 1735.7 +/- 69.1 fmol/mg protein. Specific [3H]-SQ 29,548 binding was inhibited by specific TP receptor antagonists and agonists in a rank order of potency similar to that seen in platelet membranes: SQ 33,961 much greater than SQ 29,548 greater than BM 13,505 greater than or equal to U 46619 greater than BM 13,177. PGD2, PGE2 and PGI2 did not appreciably inhibit the specific binding of [3H]-SQ 29,548. These data indicate that [3H]-SQ 29,548 binding to soluble human platelet TP receptors was specific, saturable, and reversible.


Microvascular flow vectors in normal and hypertrophic myocardium as determined by the method of colored microspheres.

Sequential in vivo infusion of two differently colored microsphere suspensions into the left atrium of normal and hypertrophic rat myocardium revealed that certain coronary capillaries contained microsphere aggregates of both colors. A capillary flow vector was established based on the sequence of colors embolized within each aggregate. Critical examination of flow vectors among neighboring capillaries enabled the characterization of capillary flow direction. Results indicated a predominance in concurrent flow direction, which decreased significantly (P less than 0.001) with capillaries further removed from an individual reference capillary. The percentage of concurrent flow was also found to be significantly lower in subendocardium (P less than 0.001) than in midmyocardium. Cardiac hypertrophy was not a contributing factor to the above findings. This study provides previously unattainable data regarding transmural capillary flow direction and suggests regional adaptations in coronary microvascular flow.


Serum levels of helper factors (IL-1 alpha, IL-1 beta and IL-6), T-cell products (sCD4 and sCD8), sIL-2R and beta 2-microglobulin in patients with B-CLL and benign B lymphocytosis.

Chronic B-lymphocytic leukemia (B-CLL) cells may be regulated by immune functions. In an attempt to analyze such functions, helper factors (IL-1 alpha, IL-1 beta and IL-6), T-cell products (sCD4 and sCD8) and sIL-2R and beta 2-microglobulin were measured in serum of patients at different stages of the disease. Patients were classified as having monoclonal lymphocytosis of undetermined significance (MLUS), stable or progressive B-CLL respectively. A significant, but modest, increase of IL-1 alpha was found in B-CLL as well as in MLUS patients whereas IL-6 levels were increased in MLUS only. sCD8 levels were increased both in MLUS and B-CLL but augmented sCD4 concentrations were found statistically significant only in progressive B-CLL. beta 2-microglobulin and sIL-2R were related to the extent of the monoclonal B-cell fraction. The data indicate an increased T-suppressor activity in both MLUS and B-CLL patients and a selective increase of helper T-cell activity in progressive B-CLL. A possible immunoregulatory influence of helper T cells on disease progression is discussed.


Improvement in outcome for very low birthweight children: apparent or real?

To determine whether improvement in the survival rate of infants with a birthweight of less than 1501 g was accompanied by an increase in the rate of neurological impairment or disability among the survivors. Two cohorts of consecutive very low birthweight infants (birthweight less than 1501 g) in one tertiary perinatal centre were followed prospectively to eight years of age; for both cohorts, comparison groups of children of birthweight more than 1501 g were randomly selected from hospital births. The first cohort was born before the introduction of assisted ventilation (1966-1970), the second after assisted ventilation was well established (1980-1982). Comparisons between cohorts, at eight years of age, of the survival rates and the rates of severe sensorineural impairments and disabilities. The survival rate for very low birthweight infants to eight years of age almost doubled between these cohorts, from 37.1% to 67.8% (odds ratio [OR], 3.4; 95% confidence interval [CI], 2.5-4.7; chi 2 = 57.6; P much less than 0.0001). The biggest gain was the increase in non-disabled survivors at eight years of age, from 52.6% in the first cohort to 80.8% in the second cohort (OR, 3.5; 95% CI, 2.2-5.7; chi 2 = 26.7; P less than 0.0001). Furthermore, the rate of severe disabilities in survivors fell substantially, from 13.6% to 4.1% (OR, 0.31; 95% CI, 0.14-0.69; chi 2 = 8.3; P less than 0.01). Of specific impairments, the rate of severe sensorineural deafness fell substantially (3.2% to 0%: OR, 0.14, 95% CI, 0.02-0.81; chi 2 = 4.8; P less than 0.05), as did the rate of severe intellectual impairment (13.0% to 2.7%: OR, 0.25; 95% CI, 0.11-0.57; chi 2 = 10.7; P less than 0.002). Only the rate of cerebral palsy increased, but not significantly (2.6% to 6.8%; OR, 2.6; 95% CI, 0.89-7.6; chi 2 = 3.0). It has been possible to improve the survival rate of very low birthweight infants over time without increasing the number of severely disabled survivors. Whether the long-term outcome for these infants is continuing to improve with more recent advances in perinatal care remains to be determined.


Effect of antiinflammatory agents on synthesis of MCP-1/JE transcripts by human blood monocytes.

The human monocyte chemoattractant protein (MCP)-1 encoded by the JE gene belongs to a family of low molecular weight secretory cytokines with monocyte-stimulating activity. JE transcripts are constitutively synthesized by normal and leukemic monocytes, as well as mesenchymal cells, including fibroblasts, vascular endothelial cells, and smooth muscle cells. Expression of MCP-1/JE is increased severalfold upon exposure of cells to recombinant human granulocyte-macrophage colony-stimulating factor but is down-regulated when cells are treated with lipopolysaccharide (LPS). Given the proinflammatory properties of MCP-1/JE, we have examined the modulatory effects of various antiinflammatory agents, including indomethacin, dexamethasone, cyclosporin A, and interleukin-4, on levels of MCP-1/JE transcripts either constitutively or inducibly expressed by human peripheral blood monocytes. Whereas indomethacin had no detectable effect on synthesis of MCP-1/JE transcripts and interleukin-4 treatment resulted in only a modest increase in steady state JE mRNA levels, exposure of monocytes to dexamethasone (DXS) led to a significant (2.5-10-fold) down-regulation of MCP-1/JE transcript levels. Studies examining the mechanism of down-regulation of JE mRNA by DXS indicated that DXS was acting transcriptionally and posttranscriptionally, by reducing the transcriptional rate of the MCP-1/JE gene and by destabilizing JE mRNA, a process requiring de novo RNA and protein synthesis. Although cyclosporin A by itself had no effect on synthesis of JE transcripts, it apparently relieved LPS-mediated down-regulation of JE transcript levels, by interfering with the destabilizing effect of LPS on JE mRNA. These results may provide new information regarding the action of antiinflammatory agents on synthesis of endogenous proinflammatory cytokines.


Increasing scientific power with statistical power.

A survey of basic ideas in statistical power analysis demonstrates the advantages and ease of using power analysis throughout the design, analysis, and interpretation of research. The power of a statistical test is the probability of rejecting the null hypothesis of the test. The traditional approach to power involves computation of only a single power value. The more general power curve allows examining the range of power determinants, which are sample size, population difference, and error variance, in traditional ANOVA. Power analysis can be useful not only in study planning, but also in the evaluation of existing research. An important application is in concluding that no scientifically important treatment difference exists. Choosing an appropriate power depends on: a) opportunity costs, b) ethical trade-offs, c) the size of effect considered important, d) the uncertainty of parameter estimates, and e) the analyst's preferences. Although precise rules seem inappropriate, several guidelines are defensible. First, the sensitivity of the power curve to particular characteristics of the study, such as the error variance, should be examined in any power analysis. Second, just as a small type I error rate should be demonstrated in order to declare a difference nonzero, a small type II error should be demonstrated in order to declare a difference zero. Third, when ethical and opportunity costs do not preclude it, power should be at least .84, and preferably greater than .90.


Nucleotide deletion of T cell receptor V gamma 2 and J gamma 2 coding sequences at V gamma 2-J gamma 2 junctions in immature B cell lines.

Immature B cell lines transformed with a temperature-sensitive mutant of Abelson murine leukaemia virus undergo preferentially V gamma 2 to J gamma 2 joinings during culture at a non-permissive temperature (39 degrees C). Here we examined nucleotide addition and deletion at the V gamma 2-J gamma 2 junctions in these V gamma 2 to J gamma 2 joinings, in comparison with the V gamma 2-J gamma 2 junctional sequences reported previously in T cells. Forty-eight V gamma 2-J gamma 2 junctions were PCR-amplified and sequenced. Only three of 48 V gamma 2-J gamma 2 junctions had nucleotide addition. The average nucleotide deletion of V gamma 2 coding sequences at the V gamma 2-J gamma 2 junctions was 6.9 nucleotides in immature B cells and 5.1 nucleotides in T cells (p less than 0.05). Also, the average nucleotide deletion of J gamma 2 coding sequences at the V gamma 2-J gamma 2 junctions was 3.1 nucleotides in immature B cells and 1.9 nucleotides in T cells (p less than 0.01). The average of total number of the deleted nucleotides at the V gamma 2-J gamma 2 junctions was 10.0 nucleotides in immature B cells and 7.0 nucleotides in T cells (p less than 0.01). No correlation was found between the extent of the nucleotide deletion of the V gamma 2 coding sequence and that of the J gamma 2 coding sequence at each V gamma 2-J gamma 2 junction in immature B and T cells. These results demonstrated that nucleotide deletion at the V gamma 2-J gamma 2 junctions was significantly wider in immature B cells than in T cells.


Isotypic and clonal variations in the interactions between model monoclonal immune complexes and the human erythrocyte CR1 receptor.

Erythrocytes (E) play a central role in handling circulating immune complexes (IC) in primates. E capture IC via complement receptors, type 1 (CR1) which can bind to C3b and C4b ligand sites generated on IC during activation of the complement cascade. The present study was designed to explore how the immunochemical properties of IC affected their interactions with human E. Model IC were constructed by combining murine monoclonal anti-dinitrophenyl (DNP) antibodies with DNP-bovine serum albumin. A panel of 10 independently-derived monoclonal IgG1, IgG2a, IgG2b, IgG3, IgM and IgA antibodies were used to construct IC and their interactions with human E were examined in vitro. The data reveal that IC constructed with the different monoclonal antibodies differed with respect to their rate of binding to E, the peak magnitude of IC binding to E, and the rate and extent of IC release from E. IC containing IgG1 antibodies (IgG1 IC), IgG2a IC, IgG2b IC, and IgA IC all bound rapidly to E, whereas IgG3 IC and IgM IC were bound relatively slowly to E. The peak magnitude of IC binding to E correlated directly with their binding rate. There was an inverse correlation between the antigen/antibody ratio of the IC and the magnitude of IC binding to E. The rate of release of the various types of IC from E also differed. IgG2a IC and IgG2b IC displayed the most rapid maximum release rates while IgG3 IC had the slowest peak release rate. IgM IC and IgA IC were also released relatively slowly from E. IgG1 IC had an intermediate release rate. There was no direct correlation between the maximum release rate and either the maximum binding rate or the peak magnitude of IC binding to E. While there were some clonotypic differences in binding and release rates between IC made with different IgG2a, IgG3 and IgM antibodies, antibody isotype appears to be of fundamental importance with respect to both the binding of IC to E and the release of IC from E. These data indicate that the immunochemical properties of IC can profoundly affect their interactions with human E and that the panel of IC constructed with monoclonal antibodies can serve as a useful model to explore these interactions.


Evidence for the presence of regional differences in the calcium antagonist receptors in lower urinary tract smooth muscle.

(+)-[3H]PN 200-100 (a dihydropyridine calcium channel antagonist) was utilized to characterize calcium channel binding sites in rabbit bladder dome, bladder base, and urethra. Specific binding of (+)-[3H]PN 200-110 to membrane particulates was saturable, reversible, linear to protein concentration, and of high affinity. The density of (+)-[3H]PN 200-110 binding sites (Bmax values in fmol/mg of protein) and the affinity constants for (+)-[3H]PN 200-110 (KD value in pM) in urethra, bladder dome and bladder base were 64.1 +/- 7.8 and 179 +/- 31; 21.9 +/- 3.0 and 213 +/- 36; and 18.8 +/- 4.2 and 140 +/- 28, respectively. Agonists and antagonists inhibited (+)-[3H]PN 200-110 binding with Ki values in the following rank order: nitrendipine less than nifedipine less than niguldipine much less than Bay K 8644 much less than verapamil. Although carbachol-induced contractile responses were 20-30 times smaller in muscle strips from urethra than from bladder base or bladder dome, KCl-induced contractions were only 3-4 times smaller in urethra than in bladder tissues. Nifedipine inhibited carbachol-induced contractions in urethra, bladder dome, and bladder base by 76%, 64%, and 60%, respectively, and completely inhibited KCl-induced contractions in all three tissues. IC50 values for nifedipine inhibition of both carbachol- and KCl-induced contractions were significantly smaller in urethra than in bladder base or bladder dome. Nitrendipine, niguldipine and verapamil inhibited urethral contractions induced by carbachol and KCl to the same degree as did nifedipine. The IC50 values, obtained from functional studies, for calcium channel antagonists were in good agreement with Ki values obtained from binding studies.(ABSTRACT TRUNCATED AT 250 WORDS)


Preferential location of N-methyl-D-aspartate (NMDA) receptors on postsynaptic membranes and on non-noradrenergic nerve terminals of the rat brain cortex.

The effect of DSP4-induced destruction of noradrenergic neurones on 3H-3-(2-carboxypiperazine-4-yl)propyl-1-phosphonic acid (3H-CPP) binding to N-methyl-D-aspartate (NMDA) receptors and on 3H-desipramine (3H-DMI) binding to the neuronal noradrenaline carrier was investigated in rat brain cortex buffy coat membranes. 3H-DMI bound with high affinity to a single site at the neuronal noradrenaline carrier (KD = 5.26 +/- 1.67 nmol/l) whereas the binding of 3H-CPP to the NMDA receptor was of intermediate affinity (KD = 274 +/- 45 nmol/l). Fourteen days after a single-dose treatment with DSP4 (1) the Bmax value for 3H-DMI binding was reduced by 74%, (2) the Bmax value for 3H-CPP binding only tended to be decreased (by 24%; not statistically significant), (3) the endogenous noradrenaline content was reduced by 70% compared to untreated controls and, (4) the absolute amount of the NMDA-evoked 3H-noradrenaline overflow but not the fractional release was reduced by 55%. It is concluded that in the rat cerebral cortex presynaptic NMDA-receptors on noradrenergic nerve endings, which have previously been detected in release experiments with NMDA on cortical synaptosomes preincubated with 3H-noradrenaline, cannot be identified in radioligand binding experiments. Obviously, the cerebral cortical NMDA receptors are predominantly located on postsynaptic neuronal membranes and potentially on non-noradrenergic nerve terminals as well.


Purification and characterization of RHP (factor H) and study of its interactions with the first component of complement.

RHP has been purified from the plasma of both normal individuals and patients with rheumatoid arthritis (RA). RHP from both these sources was shown to be identical with Factor H by reaction with antisera and N-terminal amino acid sequence analysis. Factor H, from both normal and RA sera, inhibited the solubilization of immune precipitates but did not affect prevention of immune precipitation. Factor H was shown to inhibit the haemolytic activity of fluid-phase C1, but unlike C1-inhibitor, it had little effect on C1 bound to EA (EAC1). Factor H was shown to complex with intact C1, to isolated C1q and to the C1r:C1s tetramer. However, binding of factor H to C1 did not dissociate the C1 macromolecule. A C1-Factor H complex was detected in the serum and plasma from normal individuals and patients with systemic lupus erythematosus and RA. Serum levels of this complex were reduced, by EDTA-treatment of serum and by activation of complement by the classical pathway.


Intravascular balloon dilatation therapy for intracranial arterial vasospasm: patient selection, technique, and clinical results.

Patients presenting with symptomatic intracranial arterial vasospasm are now being treated by interventional neurovascular techniques in selected cases. From a percutaneous transfemoral approach, a custom-designed silicone microballoon can be guided through the intracranial vessels, and inflated to dilate the spastic vessel(s). This technique has been useful for both focal and diffuse areas of spasm. In clinical trials, 28 patients, ranging in age from 15-73 years have been treated. A total of 99 vascular territories have been successfully dilated in both the anterior and posterior circulations. Clinical improvement following treatment was observed in 17 cases (60.7%). Technical complications directly related to therapy included 2 cases (7.1%) of vessel rupture. In long term clinical follow-up 17 patients (60.7%) had good to excellent outcome, 2 patients (7.1%) remained in poor condition, and 9 patients (32.1%) died despite therapy. In patients presenting with symptomatic intracranial arterial vasospasm who are unresponsive to medical therapy, treatment by balloon angioplasty techniques may help to improve cerebral perfusion and clinical outcome of the patient.


[Lipoprotein (a), a new atherosclerotic risk factor. Blood levels in normal pregnancy].

Serum levels of lipoprotein (a), an important factor of atherosclerotic risk, in 32 patients during normal pregnancy and puerperium, were evaluated. Our results show a significant increase of the level of this lipoprotein (M = 20 mg/dl) versus controls (M = 16 mg/dl) during all periods of pregnancy, with a very significant increase at week 20th.


Characterization of transforming growth factor-beta 1 gene expression in porcine immune cells.

We have investigated the regulation of transforming growth factor beta 1 gene expression in a variety of porcine immune cell populations, including peripheral blood mononuclear cells (PBMC), peripheral blood monocytes, alveolar macrophages and lymphoid cells from various swine lymphoid tissues. Using porcine transforming growth factor beta 1 cDNA probes in Northern blot assays, messages of 2.5 and 3.5 kb TGF beta 1 mRNA were detected in the cells investigated. A variety of mitogenic and immunomodulatory substances were examined for their ability to induce TGF beta 1 mRNA expression. These include phorbol 12-myristate 13-acetate (PMA), phytohemagglutinin (PHA), concanavalin A (Con A), lipopolysaccharide (LPS), dexamethasone (Dex), tumor necrosis factor (TNF) and interleukin (IL)-1 alpha. While low level constitutive expression of TGF beta 1 mRNA was detected from all cells investigated, PMA treatment of PBMC and alveolar macrophages resulted in a more than 10-fold increase in the steady-state level of TGF beta 1 mRNA within 2 hr of PMA addition. Also, the effect of opiate drugs, methadone (Md) and morphine (Mor), on TGF beta 1 gene expression was determined. Cells treated with opiates expressed the same levels of TGF beta 1 mRNA as untreated cells. Since TGF beta 1 biological activity can be induced by opiates, the regulation of TGF beta 1 gene expression likely involves mechanisms that do not cause changes in mRNA levels.


Differential modulation of complement factor H and C3 expression by TNF-alpha in the rat. In vitro and in vivo studies.

The biosynthesis of alternative regulatory complement protein factor H was investigated using both an in vivo rat model and an in vitro rat hepatocyte culture system, and compared to that of C3 component. Subcutaneous injection of a single dose of 20 micrograms of recombinant murine tumor necrosis factor-alpha (rmTNF-alpha) had no effect on factor H liver mRNA levels, while it increased C3 mRNA levels. In correlation with this, serum factor H levels remained unchanged after rmTNF-alpha injection, whereas C3 levels were increased. In contrast in vitro studies showed that rmTNF-alpha had no effect on factor H and C3 expression by rat hepatocytes. Recombinant human interleukin-1 alpha (rhIL-1 alpha) did not alter the expression of factor H, whereas it increased C3 expression, and recombinant human interleukin-6 (rhIL-6) stimulated expression of both proteins. This study shows that TNF-alpha is not directly responsible for the increased levels of factor H observed in vivo during induced inflammation in the rat. Its in vivo effect on C3 secretion might be secondary to the TNF-alpha-induced release of IL-1 and/or IL-6.


Effects of the thromboxane-receptor antagonists AH 23848 and BM 13.177 on human uteroplacental arteries.

We studied the effects of the thromboxane (Tx)A2-receptor antagonists AH 23848 and BM 13.177 in small isolated human uteroplacental arteries. Fetal stem villous arteries and maternal intramyometrial arteries were dissected from placental specimens and from myometrial biopsies obtained at cesarean or from nonpregnant women after hysterectomy. Vascular ring preparations were prepared and mounted in organ baths, and isometric tension was recorded. AH 23848 produced competitive, concentration-dependent inhibition of responses to the TxA2-mimic U46619 in all vessel types tested. Mean (+/- standard error of the mean) pA2 values (the negative logarithm of the concentration of antagonist needed to double the half maximum response [EC50] value for U46619) were 8.69 +/- 0.16 in the stem villous arteries, 9.58 +/- 0.33 in intramyometrial arteries from term pregnant women, and 9.25 +/- 0.47 in intramyometrial arteries from nonpregnant women. In stem villous arteries, the pA2 value for BM 13.177 was 6.15 +/- 0.13, whereas these values in intramyometrial arteries could not be assessed. However, the concentrations needed to produce inhibition of U46619-induced contractions were considerably higher for BM 13.177 than for AH 23848. Both drugs inhibited responses to prostaglandin (PG)F2 alpha and PGE2 in stem villous arteries, while leaving responses to vasopressin in intramyometrial arteries unaffected. No differences in the effects of the two antagonists were found between intramyometrial arteries from nonpregnant and term pregnant women. Our results suggest that TxA2-receptor antagonists effectively inhibit responses to TxA2 in human uteroplacental arteries, and such drugs may represent an interesting therapeutic approach in preeclampsia.


The effect of laparoscopic ablation or danocrine on pregnancy rates in patients with stage I or II endometriosis undergoing donor insemination.

The effect of treatment on stages I and II endometriosis was evaluated in 61 women with laparoscopically proven disease who were undergoing therapeutic donor insemination. Only treatment cycles completed after diagnostic laparoscopy were used for analysis. To evaluate fecundity, we performed life-table analysis on 343 treatment cycles of therapeutic donor insemination in 67 patients with stage I or II endometriosis and compared it with 212 cycles in 43 patients with no female infertility factors. Average monthly fecundity and cumulative conception rates over six cycles were calculated for each group. A significant difference was found when patients with laparoscopically proven normal anatomy were compared with those with endometriosis (P = .002). The fecundity did not differ significantly between stage I and stage II endometriosis (P greater than .05). Neither ablation during laparoscopy nor medical treatment with danocrine improved the fecundity of patients with early-stage endometriosis (P greater than .05).


[Kynurenine and its metabolites in nervous system diseases].

Kynurenine is a metabolite of the tryptophan-nicotine-amide-adenine-dinucleotide pathway. Recent preclinical and clinical data suggest that kynurenine and its metabolites (kynurenic acid, quinolinic acid) may play important role in the pathogenesis of neurological and other disorders (Huntington's disease, epilepsy, hypoxia, ulcus, hepatic and infectious diseases). Experimental data and theoretical considerations suggest that modification of kynurenine metabolism and influence of the concentrations of kynurenine metabolites may be useful therapeutic strategies in treatment of several disorders. This review is a summary of the pathobiochemical mechanisms and possible therapeutic strategies.


Standardised method of follow-up assessment of preterm infants at the age of 5 years: use of the WHO classification of impairments, disabilities and handicaps. Report from the collaborative Project on Preterm and Small for gestational age infants (POPS) in The Netherlands, 1983.

A nationwide, prospective study was initiated in The Netherlands in 1983, involving 1338 liveborn infants with a gestational age less than 32 weeks and/or a birthweight less than 1500 g. Pre- and perinatal data, methods and results of follow-up until the corrected age of 2 years have been published previously. In this paper, methods of follow-up at the age of 5 years are described. At that age, 966 children were alive, of which 927 (96%) were assessed during a home visit 2 to 6 weeks after their fifth birthday by three specially trained paediatricians. A questionnaire served to collect data on medical history, respiratory function, behaviour and socio-economic factors. Standardised tests were carried out covering the following 10 areas: congenital malformations, neuromotor function, mental development, hearing, visual function, language and speech development, behaviour, musculoskeletal system, respiratory tract and ENT problems, and growth. The outcome was recorded for separate areas and for the child as a whole using the WHO classification of impairments, disabilities and handicaps.


Beneficial effect of the ANF analog A68828 on recovery from ischemic acute renal failure.

We have recently reported that administration of the ANF analog A68828 improves renal function in an acute model of postischemic acute renal failure. The current investigation examined the question whether a short-term infusion of A68828 can attenuate the long-term decrement in renal function following an ischemic event. Acute renal failure was induced in male Sprague-Dawley rats (200-250 g) by complete occlusion of both renal arteries for 30 min. During the initial 60 min following ischemia, vehicle (0.1% BSA in saline), A68828 (10 micrograms/kg/min); dopamine (10 micrograms/kg/min); A68828 (10 micrograms/kg/min) plus dopamine (10 micrograms/kg/min); or ANF[1-28] (0.5 microgram/kg/min) were infused intravenously. In vehicle-treated animals, a very large increase in plasma creatinine was observed, with peak levels at 2 days postischemia (5.5 +/- 1.2 mg/dL). A68828 alone, A68828 with dopamine, or ANF[1-28] infusion attenuated the rise in plasma creatinine levels by approximately 50% on each day of the study; dopamine alone was no different from vehicle-treated controls. Gross histological examination of kidneys on the fourth day postischemia revealed that significantly less damage occurred only in the group treated with A68828 alone. These results indicate that infusion of a reduced-size analog of ANF for a brief period in the postischemic kidney improves renal function and lessens tissue damage as evaluated several days after the ischemic event. Furthermore, dopamine infusion provides no discernable beneficial effect.


[Disability, vacation and travel--goals of a humane travel culture].

In our society, holiday-making and travelling have become civic rights. Handicapped people however frequently find this "civic right to holiday travelling" difficult to implement in the usual manner. Mobility barriers, behavioural uncertainty in the social-communicative contact of disabled and non-disabled people but also financial limitations act as disincentives. It therefore is necessary that disabled persons' organizations, non-profit and commercial service providers but also the Federal government step-up their involvement in this field and get efforts toward improvement started, an endeavour that could be coordinated and inspired by the Tourism and People with Disabilities working group established in 1989.


[Independent living skills for physically handicapped students with profound disability].

Traditional educational curricula are inadequate for physically disabled students with very severe consequences of disablement, as they are insufficiently geared toward building Independent Living skills. Typical of their post-school situation nowadays, only few of them are in a position of making a direct transition into vocational training. The majority continues to need intensive educational-therapeutic services. The schools for the physically disabled seek to smoothen their transition into adult life by leaving-grade programmes aimed at providing opportunities for work-related experience as well as at building coping skills for day-to-day living. The work and life preparation programme represents a special organizational and didactical conception in this context, where the demands of adult life serve as the starting point for developing the learning and educational goals to be attained.


[Patients with severe multiple handicaps in workshops for the handicapped--a project of the Berlin Workshops for the Handicapped GmbH (BWB) 4 December 1989 to 31 May 1990].

A project had been carried out at Berliner Werkstätten für Behinderte GmbH (BWB) aimed at integrating cerebral palsied persons with very severe, multiple disability in a workshop for the disabled. Client workshop readiness had been determined on the criteria of social capacity, extraordinary care need at the workplace, and economic viability of work. The project, planned to last 6 months, had to be discontinued because the 9 disabled persons included in this programme refused to cooperate after a short period of time.


[Analysis of the capacity of handicapped patients in comparison with non-handicapped patients].

A catalogue of psychological items has been developed that enables any disabled person's job-related abilities as well as the psychological requirements of any job to be assessed. With this newly developed instrument, ability profiles of slow learners and of non-disabled persons have been drawn up. Structural analysis of these data show specific differences in the ability structure of the two groups. The results indicate new ways for well-directed, disability-related promotion in vocational rehabilitation.


New antagonists of the vasopressin receptor mediated contraction in rat tail artery.

A perfused isolated rat tail artery preparation was employed to study antagonistic properties of four newly synthesized arginine-vasopressin (AVP) analogues against the V1 receptor. The activity of the agents SCATyr(Me)AVP, OCATyr(Me)AVP, OCAAVP and SCAAVP was related to that of a recognized antagonist d(CH2)5Tyr(Me)AVP. SCATyr(Me)AVP elicited outstanding antagonistic properties by blocking at concentration of 10(-7) M nearly completely the constrictory activity of AVP. At concentration of 10(-9) M the agent inhibited the AVP-induced constriction of artery about 40 times more effectively than the oxytocin (OXT)-induced constriction. The results obtained prove the validity of the structure-activity relationship based search for new potent V1 receptor antagonists.


[The course of rehabilitation in patients with locked-in syndrome].

On the basis of observations of the course taken by 5 patients with so-called Locked-in Syndrome (LIS), the level of functional performance that can be achieved by long-term rehabilitation notwithstanding very severe initial symptoms is set out. The priority measures in the early phases of the condition and the particular problems of long-term rehabilitation are summarized in a checklist. Technical aids to ameliorate communication are dealt with in some detail. Special importance is attributed to patient-centered psychotherapeutic support and to family involvement.


Fusion of the leucine zipper gene HLF to the E2A gene in human acute B-lineage leukemia.

A t(17;19) chromosomal translocation in early B-lineage acute leukemia was shown to result in chimeric transcripts that contain sequences from the E2A basic helix-loop-helix transcription factor gene on chromosome 19, fused to sequences from a previously unidentified gene (HLF) on chromosome 17 that encodes a hepatic leukemia factor. The chimeric protein consisted of the amino-terminal transactivation domain of E2A linked to the carboxyl-terminal basic region-leucine zipper domain of HLF. HLF was normally expressed in liver and kidney, but not in lymphoid cells, and was found to be closely related to the leucine zipper-containing transcription factors DBP (albumin D-box binding protein) and TEF (thyrotroph embryonic factor), which regulate developmental stage-specific gene expression.


Laparoscopic cholecystectomy in a community hospital setting.

Recently, laparoscopic cholecystectomy has become the preferred surgical procedure for removal of the gallbladder. However, many surgeons believe that the safety and efficacy have yet to be proved in the community hospital setting. To address this concern, a retrospective chart review of the initial 271 instances of inpatient laparoscopic cholecystectomy within a community hospital was undertaken. All procedures were performed by 15 general surgeons in private practice and residents in general surgery. Of the 271 patients, 11 were converted to open cholecystectomy. Surgical complications occurred in six of the 260 instances of laparoscopic cholecystectomy (2.3 percent), with only one injury to the common bile duct. Major postoperative complications occurred in 23 patients, including severe postoperative pain (nine patients), prolonged ileus (seven patients), bile leakage (three patients), retained common duct stones (two patients), respiratory failure (one patient) and postoperative myocardial infarction (one patient). The period of hospitalization ranged from one to 64 nights with a median of one night. The operative mortality rate was zero percent. Multivariate analysis identified two factors associated with an increased risk of postoperative complications. Patients 70 years of age or older and patients whose operating times were greater than one hour and 45 minutes were at increased risk for postoperative complications. We believe that these data represent the general outcomes of the laparoscopic procedure in a community hospital setting and lend support to the argument that the procedure can be performed safely and effectively in this setting.


Effect of GR32191 and other thromboxane receptor blocking drugs on human platelet deposition onto de-endothelialized arteries.

A range of thromboxane A2 receptor blocking (TxRB) drugs, prostacyclin and aspirin have been assessed as inhibitors of human platelet deposition onto rabbit and human de-endothelialized arteries in vitro. Platelet deposition was quantified by measuring the radioactivity associated with de-endothelialized arteries following superfusion with 111indium-labelled human platelets reconstituted in blood. Using rabbit aorta, all of the compounds tested produced a similar maximum inhibition (approximately 70%) of platelet deposition; from scanning EM studies the residual deposition appeared to represent a monolayer of adhered platelets. The potency of the TxRB's for inhibiting deposition was GR32191 greater than or equal to GR36246 greater than SQ29,548 greater than ICI185282 greater than or equal to AH23848 much greater than BM13.177 consistent with their TxRB potency on human platelets. Using human umbilical arteries, the TxRB's achieved a smaller maximum inhibition of deposition (approximately 50%) than did prostacyclin or the fibrinogen receptor blocking peptide Gly-Arg-Gly-Asp-Ser (GRGDS) (60-75%). In addition, using human umbilical arteries, the structurally-related TxRB's GR32191 and GR36246 exhibited a greater than 1000-fold enhancement in potency as inhibitors of platelet deposition over that seen in the rabbit aorta. In preliminary experiments, GR32191 also displayed a similar high potency on human cerebral arteries. In contrast, the structurally unrelated compounds SQ29,548, ICI185282 and BM13.177 exhibited similar potencies on human umbilical arteries to those observed on the rabbit aorta; aspirin and prostacyclin also displayed similar potencies on the two preparations. The enhanced effect of GR32191 and GR36246 on human umbilical arteries therefore appears unrelated to their action as TxRB's on human platelets although the mechanism of this unique action is at present unknown. However, if these drugs exhibited a similar high potency for preventing mural thrombus formation in vivo in man, they may represent a major advance in the treatment of occlusive vascular disease.


Plasmin stimulates the release of dermatan sulfate from vascular smooth muscle cells in culture.

We investigated the effect of plasmin on the release of sulfated glycosaminoglycans (GAG) from cultured vascular smooth muscle cells. Confluent cultures of vascular smooth muscle cells from bovine aorta were labelled with [35S]sulfate and incubated at 37 degrees C for principally 60 min in a serum-free medium in the presence of plasmin. Plasmin at 10 mU/ml (approximately 2.7 micrograms/ml) and above significantly increased the release of [35S]sulfate-labeled GAG (35S-GAG) from the cell layer after a 60 min incubation. A time course study showed that plasmin at 10 mU/ml significantly increased the 35S-GAG release after 20 min and longer. However, plasminogen at 100 mU/ml and below did not cause a significant change of the 35S-GAG release after a 90 min incubation. A characterization of 35S-GAG revealed that plasmin increased both dermatan sulfate and the other 35S-GAG in the medium. Plasmin at 100 mU/ml enhanced the cell detachment significantly but only slightly. From these results, it was suggested that a endogenous thrombin inhibitor heparin cofactor II may be activated in the liquid phase by dermatan sulfate released from plasmin-stimulated vascular smooth muscle cells when the vascular is disrupted and plasma is exposed to extravessel.


Photodynamic therapy.

The preliminary data suggest that red-light whole-bladder photodynamic therapy is safe and effective in the treatment of Tis and may be useful in the prophylactic management of superficial bladder cancer. Theoretically, whole-bladder photodynamic therapy has the advantage of higher efficacy after a single treatment than most conventional modalities for superficial bladder cancer. In patients with Tis, the complete response rate is 88%, and 25% have recurrences during a mean follow-up of 20 months (range 12-60). In patients undergoing prophylaxis, the recurrence rate is 31% and the median time to recurrence is 18 months. Importantly, none of the high-risk patients treated with whole-bladder photodynamic therapy has developed disease progression in stage or grade at the time of recurrences. Whole-bladder therapy also has the potential advantage of repeat treatment without increased tumor resistance or increased morbidity. Data from the present phase II-III clinical trials involving a large number of patients will define the role of photodynamic therapy in the management of superficial bladder cancer.


[The cystic duct stump after laparoscopic cholecystectomy].

The aim of this study was to evaluate the length of cystic-duct stumps after laparoscopic cholecystectomy. 113 patients underwent intravenous cholangiography 2 to 3 months postoperatively, whereby a cystic-duct remnant of up to 1 cm was found in 34.5% (n = 39) and between 1 and 2 cm in 36.3% (n = 41). In 24.8% (n = 28) a stump measuring 2 to 3 cm was registered and in 4.4% (n = 5) the cystic-duct remnant was more than 3 cm. Possible consequences with regard to the development of the postcholecystectomy syndrome are discussed. In our experience laparoscopic cholangiography seems to be a useful procedure to detect cysticolithiasis and as preliminary measure to undertaking the correct therapeutic steps. Adherence of this strategy might avoid possible later complications by elimination of their principal cause.


Human helper T cell lines established by coculture of normal human cord leukocytes with an HTLV-II-infected rabbit leukocyte cell line (Ra-IIA).

Three helper T cell lines, designated CR-IIA (CR-IIA-1, CR-IIA-2, and CR-IIA-3), were established by coculturing normal human cord leukocytes with a lethally irradiated HTLV-II (human T-lymphotropic virus type II)-infected rabbit leukocyte cell line (Ra-IIA). CR-IIA had a normal human karyotype and expressed the surface markers CD3(+), CD4(+), CD8(-), CD19(-), CD25(+) and HLA-DR(+), confirming their helper T cell nature. CR-IIA cells were all free of Epstein-Barr virus nuclear antigen and were immunoreactive with serum samples from HTLV-I- or HTLV-II-infected patients and with anti-HTLV-I, p19 or p24 antibody. The provirus genome of HTLV-II was detected in these cell lines by the polymerase chain reaction combined with a digoxigenin-enzyme-linked immunosorbent assay. Electron microscopy of CR-IIA-1 cells revealed a few immature type C virus particles. These results suggest that HTLV-II was transmitted from the infected rabbit leukocyte cell line to human cord helper T lymphocytes with the development of immortalized HTLV-II-producing T cell lines.


External calcium sensitivity of low sodium contractures in the control and hypertrophied right ventricle of the ferret.

The existence of possible differences of calcium (Ca2+) fluxes through the sarcolemmal sodium-calcium (Na+/Ca2+) exchanger during hypertrophy has been tested by comparing the characteristics of the contracture--as an indicator of the intracellular Ca2+ concentration--induced by partial or total withdrawal of external sodium (Na+), in the absence of external potassium, in the right ventricular trabeculae of adult ferret hearts. Pressure-overload was induced by pulmonary artery clipping and led to an increase of the right ventricular weight of 60%. At an external Ca2+ concentration ([Ca2+]o) of 3 mM, the dependence of the contractures on extracellular sodium concentration ([Na+]o), the rate of tension development, the time course of spontaneous relaxation and the time course for the repriming of the contracture were unchanged by hypertrophy. However, the relationship between [Ca2+]o and contracture amplitude at various [Na+]o showed that the apparent affinity of the contracture for [Ca2+]o was decreased in hypertrophied preparations. Thus, in 0 mM [Na+]o, half-maximal contracture was induced at a [Ca2+]o of 0.012 +/- 0.016 mM and 0.171 +/- 0.021 mM in control (n = 11) and hypertrophy (n = 12) respectively (P less than 0.001). Although these data may be indicative of a decreased Ca2+ influx through the Na+/Ca2+ exchanger, it cannot be excluded that intracellular buffering mechanism may also be involved in this differential response to [Na+]o withdrawal.


Postural changes in atrial natriuretic peptide (ANP) and fluid balance in conscious goats.

To gain insight into how weak physiological stimuli suffice to release ANP, the effects of 15 degrees head-up and head-down tilt, 90 min each in succession, on plasma atrial natriuretic peptide (ANP), plasma cortisol, plasma and urinary Na, K and osmolality, plasma proteins and haematocrit were studied in five conscious goats before and after 2 wk daily training for the tilting procedure. In the trained goats the 15 degrees tilt did not affect the plasma ANP, cortisol or the urine excretion significantly. Total plasma proteins decreased significantly. In the untrained goats, on the other hand, an increasing trend was observed in the plasma ANP and cortisol as well as in the urine Na excretion during the head-up tilt. During the head-down tilt, the levels of ANP and Na excretion remained elevated. The plasma ANP was significantly increased after 40 min, by 28% as compared to the pre-tilting level. The plasma cortisol was first elevated, but then returned to the starting level. The results suggest that in the trained goats the responses to tilting were unmasked. Despite minor effects of the 15 degrees tilt in itself, increased plasma ANP seemed to be associated with increased natriuresis.


[Laparoscopic ureterolithectomy. A new option].

An integral conception of patients with various diseases can benefit from the contemporary technological developments which allow to differentiated non-invasive approaches as diagnostic and therapeutical resources. This kind of patient has allowed further development of laparoscopic surgery in urology derived from the addition of reported conditions: obstructive prostate carcinoma, juxta-vesical gross lithiasis imbedded for 2 years, complexity of ureteroscopic access, large volume for extracorporeal lithotripsy and the need to know the extend of nodular affectation.


Conservation of docosahexaenoic acid in the retina.

Over the last several years, evidence has accumulated that n-3 fatty acids, particularly 22:6n-3, are essential for the development of the structure and function of the visual system. The importance of 22:6n-3 is reflected in the tenacious manner in which the retina conserves this fatty acid during n-3 deficiency. We have shown that conservation is achieved by recycling 22:6n-3 within the retina or between the retina and the pigment epithelium. Within the retina, recycling could be accomplished by deacylation-reacylation reactions (Louie et al., 1991; Zimmerman and Keys, 1988). Recycling between the retina and the RPE may be achieved through specific transport proteins, possibly interphotoreceptor retinoid-binding protein (Bazan et al., 1985) and/or apolipoprotein E (Bazan et al., 1991).


Docosahexaenoic acid uptake and metabolism in photoreceptors: retinal conservation by an efficient retinal pigment epithelial cell-mediated recycling process.

After 18:3 omega 3 is obtained from the diet, it is accumulated by the liver, where it is esterified and temporarily stored as triacylglycerols. As it is required, 18:3 omega 3 is elongated and desaturated to 22:6 omega 3, then released into the circulation with lipoprotein carriers. RPE cells remove the 22:6 omega 3 from the choriocapillaris and subsequently release it to the retina proper. In the frog, all 22:6 omega 3 input to the photoreceptors occurs by way of the RPE cells. After passing through the interphotoreceptor matrix, it is selectively taken into the myoid region of photoreceptor cells where it is immediately activated and esterified onto position 2 (and sometimes also position 1) of a glycerol molecule. Some phospholipids are passed through the endoplasmic reticulum and Golgi apparatus, while others are not. Generally, transport to the outer segments seems to be independent of the Golgi apparatus. Addition to rod outer segments occurs in two ways: i) a general diffuse pathway, probably common to all fatty acids, which rapidly labels the entire outer segment; and ii) a specific dense pathway, utilized only by 22:6 omega 3-containing phospholipids, which become locked into the matrix of disc membranes along with opsin. There appears to be no exchange between these two forms of label. Accumulation of newly synthesized basal discs pushes older, 22:6 omega 3-laden discs apically until the outer segment tips, high in 22:6 omega 3-phospholipids (the dense form of outer segment label), are shed into the RPE cytoplasm. There, as the 22:6 omega 3 fatty acids are released from the disc membranes during degradation, a recycling mechanism immediately directs these essential fatty acids back into the interphotoreceptor matrix, thus conserving this molecule in the retina, and permitting it to be again selectively taken up by the photoreceptors for photomembrane synthesis. The process of 22:6 omega 3 handling and trafficking by the retina is specifically orchestrated around a conservation mechanism that is regulated by the RPE cells and that ensures, through a short feedback loop from the phagosomes to the interphotoreceptor matrix, adequate levels of 22:6 omega 3 for photoreceptors at all times.


NMDA receptor-mediated arachidonic acid release in neurons: role in signal transduction and pathological aspects.

The N-methyl-D-aspartate (NMDA)-sensitive subtype of glutamate receptor, which gates Ca(2+)-permeable ion channels, is known for its role in learning and memory formation, in the induction of long-term potentiation, and also in seizure activity and neurotoxicity. In primary cultures of cerebellar neurons, agonists of NMDA receptors induce a dose-dependent release of [3H]arachidonic acid ([3H]AA), which is potentiated by activation of the glycine-positive modulatory site and inhibited by NMDA receptor antagonists. NMDA receptor-induced [3H]AA release is inhibited by quinacrine and partially depends on the presence of extracellular calcium. The [3H]AA release is not sensitive, however, to pretreatment with pertussis or cholera toxin, which suggests a Ca(2+)-dependent activation of phospholipase A2 not employing G proteins. Pretreatment of cultures with the natural and semisynthetic sphingolipids GT1b and PKS 3, respectively, inhibits NMDA receptor-mediated [3H]AA release. We also demonstrated glutamate-evoked [3H]AA release from rat hippocampal slices, which is NMDA receptor mediated, calcium dependent and sensitive to quinacrine. Arachidonic acid and its metabolites have been shown to play a role as second messengers and to modulate neuronal activity. Moreover, they are thought to act as transsynaptic modulators in the mechanism of NMDA receptor-induced long-term potentiation in the hippocampus. Their role in ischemic brain pathology has also been postulated. Our experiments on cultured cerebellar granule cells, incubated in a Mg(2+)-free medium deprived of glucose and oxygen, demonstrated a time-dependent stimulation of [3H]AA release. This release was inhibited by antagonists of NMDA receptors and by quinacrine. Stimulation of NMDA-sensitive glutamate receptors and the subsequent calcium-mediated activation of phospholipase A2 may play a role in the in vivo release of arachidonic acid during brain ischemia. This hypothesis is supported by the observation that the enhanced level of thromboxane B2 in the gerbil brain after 5 min of global ischemia is reduced by the systemic application of either the NMDA antagonist MK-801 or the ganglioside GM1.


Left ventricular hypertrophy and antihypertensive therapy.

Left ventricular hypertrophy is more common in hypertensive individuals than in normotensive persons. Its presence in hypertensive patients is associated with an increased incidence of ventricular arrhythmias, myocardial infarction, congestive heart failure, stroke and cardiovascular mortality. Echocardiography is more sensitive than electrocardiography in detecting left ventricular hypertrophy. Echocardiographic evidence of this condition in patients with borderline hypertension may identify those who need treatment. Weight reduction and drug therapy can prevent or reverse ventricular hypertrophy in hypertensive patients. Recent studies suggest that some antihypertensive drugs are more effective than others in reducing left ventricular hypertrophy. These agents include beta-adrenergic blockers, angiotensin converting enzyme inhibitors, calcium channel blockers and sympatholytic agents. Although little evidence exists to show that reduction of left ventricular mass decreases cardiovascular morbidity and mortality, avoidance of antihypertensive agents that may aggravate hypertrophy would seem prudent.


The effect of late patency of the infarct-related coronary artery on left ventricular morphology and regional function after thrombolysis.

The use of early coronary angiography to assess the benefits of coronary patency on left ventricular size and function fails to account for subsequent reocclusion or spontaneous reperfusion. To investigate the relationship between late vessel patency and changes in left ventricular structure and function after thrombolysis, echocardiography was performed within 48 hours and at 6 to 12 weeks in 30 patients treated with intravenous thrombolysis. Left ventricular endocardial surface area index (ESAj; cm2/m2) and extent of abnormal wall motion were quantitated in those with a patent (n = 20) and those with an occluded (n = 10) infarct-related artery on coronary angiography performed 8 +/- 6 days after thrombolysis. Mean ESAi increased from (53 +/- 7 to 61 +/- 10 cm2/m2; p less than 0.02) in the occluded group during the follow-up period but remained unchanged (60 +/- 11 to 62 +/- 11 cm2/m2; p = NS) in the patient group. Mean percentage of abnormal wall motion decreased in the patent group (27 +/- 16% to 18 +/- 16%; p less than 0.01), whereas no significant change was noted in the occluded group (20 +/- 13% to 23 +/- 17%; p = NS). Thus coronary patency at days after thrombolysis is associated with both improvement in regional left ventricular function and attenuated left ventricular dilatation.


Excimer laser angioplasty in the atherosclerotic rabbit: comparison with balloon angioplasty.

The early and late results of excimer laser angioplasty and balloon angioplasty were compared in atherosclerotic rabbit iliac arteries. Immediately after laser angioplasty (n = 13) with a bare 600 microns fiber, there was a 33% increase in angiographically measured minimum lumen diameter; after balloon angioplasty (n = 12), there was a 53% increase. Restenosis (defined as loss of at least 50% of the gain achieved by angioplasty) occurred in none of six laser-treated rabbits studied 1 month later, compared with four of six balloon-treated rabbits (p = 0.06). Planimetric measurements of cross sections of the arterial wall 1 month after angioplasty showed less intimal and medial tissue in laser-treated (1.8 +/- 0.2 mm2) than in the balloon-treated rabbits (3.0 +/- 0.4 mm2; p less than 0.05). Typical thermal effects were absent on microscopic examination of laser angioplasty sites. It is concluded that in this animal model, excimer laser irradiation results in an immediate increase in lumen diameter comparable with balloon angioplasty, but is associated with less residual atheromatous tissue than balloon angioplasty and a trend toward a lower rate of restenosis.


Nonoperative treatment of membranous obstruction of the inferior vena cava by percutaneous balloon transluminal angioplasty.

Percutaneous transluminal angioplasty (PTA) was performed with the Inoue balloon catheter in nine patients with membranous obstruction of the inferior vena cava (MOVC) between November 1988 and April 1991. There were five men and four women, aged 30.8 +/- 8.5 years. Two patients had had previous surgical treatment. Three patients had complete and six had incomplete MOVC. The caval pressure below the MOVC was 24.0 +/- 5.6 and 11.8 +/- 5.3 mm Hg (p less than 0.0001) before and after PTA, respectively. The caval diameter at the site of MOVC increased from 1.5 +/- 1.7 to 20.3 +/- 2.6 mm (p less than 0.0001), and the maximal caval diameter below the MOVC decreased from 28.7 +/- 12.9 to 19.8 +/- 9.9 mm (p = 0.006), before and after PTA, respectively. One patient died of massive pulmonary embolism following successful PTA. All the other eight patients remained asymptomatic during an 18.5 +/- 11.5 months (range 3.5 to 32) follow-up period. Two-dimensional ultrasonograms showed no recurrence of MOVC. We conclude that PTA with the Inoue balloon catheter is an effective and safe alternative to surgical treatment of MOVC.


Changes in left ventricular size, wall thickness, and function in anemic patients treated with recombinant human erythropoietin.

Left ventricular size and function were evaluated in 15 anemic chronic hemodialysis patients before and after the administration of recombinant human erythropoietin (rHuEPO). All patients were studied with two-dimensional and M-mode echocardiographic examinations before the initiation of rHuEPO (T1) and at 28 +/- 7 weeks of rHuEPO therapy (T2). The two-dimensional targeted M-mode echocardiographic measurements obtained were: end-diastolic dimension (EDD); end-systolic dimension (ESD); stroke dimension (SD); dimensional shortening (SD/EDD); systolic posterior wall thickness (PWs); diastolic posterior and interventricular septal thickness; end-systolic wall stress (ESWS); and left ventricular mass. Mean hematocrit in these patients increased almost 50%. The EDD decreased from a mean value (+/- SEM) of 6.41 +/- 0.33 to 4.93 +/- 0.21 cm (p less than 0.05). ESD decreased from a mean value of 4.16 +/- 1.2 to 2.77 +/- 0.06 cm (p less than 0.05). The calculated mean SD decreased slightly but not significantly from 2.21 +/- 0.69 to 2.19 +/- 0.60 cm. The calculated SD/EDD increased from a mean 0.35 +/- 0.09 to 0.44 +/- 0.07 (p less than 0.05). ESWS fell from 59.2 +/- 12.2 to 37.6 +/- 9.3 gm/cm2 (p less than 0.01), and left ventricular mass fell (p less than 0.05) from 347 +/- 15.2 to 227 +/- 59 gm. There was no significant difference in resting heart rate or systolic blood pressure between T1 and T2. The increase in dimension shortening reflects afterload reduction, as indicated by the fall in end-systolic wall stress.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of weight reduction on blood lipids and lipoproteins: a meta-analysis.

Studies designed to examine effects of weight reduction by dieting on total cholesterol (TC), low-density-lipoprotein cholesterol (LDL-C), high-density-lipoprotein cholesterol (HDL-C), very-low-density-lipoprotein cholesterol (VLDL-C), and triglycerides (TGs) have reported inconsistent results. The purpose of this study was to quantify effects of weight loss by dieting on lipids and lipoproteins through the review method of meta-analysis. Results from the 70 studies analyzed indicated that weight reduction was associated with significant decreases (P less than or equal to 0.001) and correlations (P less than or equal to 0.05) for TC (r = 0.32), LDL-C (r = 0.29), VLDL-C (r = 0.38), and TG (r = 0.32). For every kilogram decrease in body weight, a 0.009-mmol/L increase (P less than or equal to 0.01) in HDL-C occurred for subjects at a stabilized, reduced weight and a 0.007-mmol/L decrease (P less than or equal to 0.05) for subjects actively losing weight. Our results indicate that weight reduction through dieting can be a viable approach to help normalize plasma lipids and lipoproteins in overweight individuals.


Cooperative activation of myosin by light chain phosphorylation in permeabilized smooth muscle.

The purpose of this study was to determine the quantitative relationship between the number of myosin molecules that increase their ATPase activity and the degree of myosin light chain phosphorylation in smooth muscle. Single turnover experiments on the nucleotide bound to myosin were performed in the permeabilized rabbit portal vein. In the resting muscle, the rate of exchange of bound nucleoside diphosphate was biphasic and complete in approximately 30 min. When approximately 80% of the myosin light chain was thiophosphorylated, the nucleoside diphosphate exchange occurred at a much faster rate and was almost complete in 2 min. Thiophosphorylation of 10% of the myosin light chains caused an increase in the rate of ADP exchange from much more than 10% of the myosin subfragment-1. Less than 20% thiophosphorylation of the total myosin light chains resulted in the maximum increase in ADP exchanged in 2 min. It appears that a small degree of myosin light chain phosphorylation cooperatively turns on the maximum number of myosin molecules. Interestingly, even though less than 20% thiophosphorylation of the myosin light chain caused the maximum exchange of ADP within 2 min, higher degrees of thiophosphorylation were associated with further increases in the ATPase rates. We conclude that a small degree of myosin light chain thiophosphorylation cooperatively activates the maximum number of myosin molecules, and a higher degree of thiophosphorylation makes the myosin cycle faster. A kinetic model is proposed in which the rate constant for attachment of unphosphorylated cross bridges varies as a function of myosin light chain phosphorylation.


Response of slow and fast muscle to hypothyroidism: maximal shortening velocity and myosin isoforms.

This study examined both the shortening velocity and myosin isoform distribution of slow- (soleus) and fast-twitch (plantaris) skeletal muscles under hypothyroid conditions. Adult female Sprague-Dawley rats were randomly assigned to one of two groups: control (n = 7) or hypothyroid (n = 7). In both muscles, the relative contents of native slow myosin (SM) and type I myosin heavy chain (MHC) increased in response to the hypothyroid treatment. The effects were such that the hypothyroid soleus muscle expressed only the native SM and type I MHC isoforms while repressing native intermediate myosin and type IIA MHC. In the plantaris, the relative content of native SM and type I MHC isoforms increased from 5 to 13% and from 4 to 10% of the total myosin pool, respectively. Maximal shortening velocity of the soleus and plantaris as measured by the slack test decreased by 32 and 19%, respectively, in response to hypothyroidism. In contrast, maximal shortening velocity as estimated by force-velocity data decreased only in the soleus (-19%). No significant change was observed for the plantaris.


Differences in regulation between nuclear and cytoplasmic Ca2+ in cultured smooth muscle cells.

The free Ca2+ concentrations in the nucleus ([Ca2+]n) and cytoplasm ([Ca2+]c) of cultured smooth muscle cells were estimated using the fluorescent dye indo-1 and the ACAS 570 confocal laser microscope. In resting DDT1MF2 smooth muscle cells [Ca2+]n was found to be lower than [Ca2+]c. Both values increased transiently in response to histamine (100 microM), but during this stimulation [Ca2+]n exceeded [Ca2+]c. Maximal increase of [Ca2+]n was observed in the center of the nucleus, and a maximal increase of [Ca2+]c was observed in the immediate vicinity of the plasma membrane. A similar response was obtained with other agonists, such as carbachol or ATP. Comparable results with ATP were obtained in cultured aorta cells. The differential rise of [Ca2+]n over [Ca2+]c in DDT1MF2 cells did not occur during either spontaneous release of Ca2+ or Ca2+ release induced by caffeine (7.5 mM). The differential rise during histamine stimulation was abolished by the presence of the intercalating substance ethidium bromide. Thapsigargin, a presumed specific inhibitor of the endoplasmic reticulum Ca(2+)-Mg(2+)-adenosine-triphosphatase, abolished the Ca2+ gradient between nucleus and cytosol at rest. During subsequent histamine stimulation the Ca2+ increase was largely blocked in both compartments and attained similar levels. We propose that the lower value of [Ca2+]n at rest is dependent on an active Ca2+ extrusion system. The differential rise of [Ca2+]n over [Ca2+]c during agonist stimulation can be explained by an influx of Ca2+ from perinuclear stores and/or by a release of intranuclear Ca2+ possibly mediated by a process dependent on the inositol lipid metabolism.


Alloxan, but not streptozotocin, increases blood perfusion of pancreatic islets in rats.

It has recently been shown that selective B-cell toxins alloxan and streptozotocin (STZ) possess marked effects also on the vascular system. To evaluate to what extent changes in blood perfusion of islets induced by alloxan or STZ could be of importance for diabetogenic action of these compounds, we first investigated acute effects of alloxan (75 mg/kg body wt iv) and STZ (40 mg/kg body wt iv) on both whole pancreatic blood flow (PBF) and islet blood flow (IBF) in adult rats. Alloxan caused a marked increase in IBF, which was most pronounced 3 min after administration and remained for 30 min. PBF, however, was decreased 3 min after alloxan administration but was similar to that of control animals from 10 min and onward. These two opposite effects on IBF and PBF caused the fraction of whole PBF diverted through islets to increase from approximately 10 to 50%. Pretreatment with glucose (2 g/kg body wt iv), indomethacin (3.5 mg/kg body wt iv), dimethyl sulfoxide (10 ml/kg body wt ip of a 33% solution), superoxide dismutase (SOD, 1,000 kU/kg body wt iv), NG-methyl-L-arginine (30 mg/kg body wt iv), theophylline (7 mg/kg body wt iv), or terbutaline (1 mg/kg body wt iv) failed to affect stimulation of IBF by alloxan observed at 3 min. SOD was found to exert a marked stimulation of IBF both when given alone and together with alloxan. Alloxan increased IBF and decreased PBF also in a syngeneic pancreaticoduodenal graft in rats but did not affect flow distribution in a perfused pancreas-duodenum preparation.(ABSTRACT TRUNCATED AT 250 WORDS)


cDNA sequence and tissue expression of bovine vacuolar H(+)-ATPase M(r) 70,000 subunit.

cDNA clones encoding the 70,000 relative molecular weight (M(r)) subunit of the bovine vacuolar proton-adenosine triphosphatase (H(+)-ATPase) were isolated, and a total length of 3.1 kb in overlapping clones from bovine kidney and brain libraries was sequenced. The cDNA contains a 1.9-kb coding region and yields a deduced protein sequence of 618 amino acids. The subunit sequence has 50% amino acid identity with the corresponding subunit from yeast, carrot, and Neurospora vacuolar H(+)-ATPases. The internal regions of the protein are most highly conserved, whereas the NH2- and COOH-terminals exhibit variability. mRNA levels of the M(r) 70,000 subunit were examined in multiple bovine tissues and were found to be expressed at highest levels in kidney medulla and cortex, at moderate levels in brain and adrenal gland, and at low levels in liver, muscle, and heart.


Localization of mRNAs coding for isozymes of plasma membrane Ca(2+)-ATPase pump in rat kidney.

We have studied localization of mRNAs coding isozymes of rat plasma membrane Ca(2+)-adenosinetriphosphatase pump (rPMCA) in the rat kidney, with use of reverse transcription (RT) with subsequent amplification by polymerase chain reaction (PCR). When zones of the kidney were separated by macrodissection, a large amount of mRNA coding isozyme rPMCA1 was found in all zones; mRNA for isozyme rPMCA2 was abundant in cortex and in outer medulla, and mRNA for isozyme rPMCA3 was prominent in outer medulla. The mRNAs were analyzed in microdissected cortical nephron segments by use of RT-PCR approach described previously [T. Moriyama, H. R. Murphy, B. M. Martin, and A. Garcia-Perez. Am. J. Physiol. 258 (Renal Fluid Electrolyte Physiol. 27): F1470-F1474, 1990]. We detected mRNA for isozyme rPMCA2 in microdissected distal convoluted tubules (DCT) and in cortical thick ascending limbs (CTAL) and, less consistently, also in proximal convoluted tubule and in glomeruli. The mRNA for isozyme rPMCA1 was abundant in glomeruli but was absent in all examined cortical tubular segments. Our results document that mRNAs for all three major isozymes of rPMCA are present and show a unique distribution in the three major zones of rat renal parenchyma. Specific mRNA coding for rPMCA2 was detected in cortical tubules, namely in CTAL and DCT, whereas mRNA coding isozyme rPMCA1 was found in glomeruli. We suggest that isozyme rPMCA2 might be specifically related to epithelial cells and their function, whereas rPMCA1 is probably a component of nonepithelial cells including these in glomeruli.


Left ventricular diastolic and systolic performance during chronic experimental aortic regurgitation.

To study the time course of left ventricular structural and functional responses to chronic aortic regurgitation, aortic regurgitation was surgically induced in rabbits, and Doppler echocardiography was performed preoperatively and serially postoperatively for up to 2.5 yr. Twenty-five New Zealand White rabbits underwent surgical induction of aortic regurgitation and 13 control animals underwent sham operation. Left ventricular endocardial and epicardial surfaces were digitized from M-mode echocardiograms to measure the rates of change of cavity dimensions and wall thicknesses during diastolic relaxation and systolic contraction. Aortic regurgitant animals developed left ventricular dilatation and eccentric hypertrophy that remained relatively stable throughout the follow-up period. Compared with baseline values, left ventricular mass increased 120% and left ventricular internal dimension at end diastole increased 40%, whereas posterior wall thickness at end diastole and fractional shortening remained relatively stable. Left ventricular diastolic performance was enhanced at 6 mo after operation, a finding associated with increased volume load and heart rate following induction of aortic regurgitation. Diastolic performance was then reduced at 12 mo after operation and demonstrated no further decline throughout the remainder of the follow-up period. In contrast, left ventricular systolic performance was not altered following operation and remained preserved until the final assessment at up to 2.5 yr. Thus alterations in diastolic performance occurred without impairment of systolic performance during long-term follow-up of chronic experimental aortic regurgitation.


Are there clinical differences between familial and nonfamilial Alzheimer's disease?

The purpose of the study was to determine whether there are differences in clinical characteristics in two groups of patients with Alzheimer's disease, those reported to have a family history of dementia and those without a family history of dementia. Using a data set from an Alzheimer's disease patient registry, funded as part of a National Institute on Aging cooperative agreement, the authors made comparisons of sociodemographic and clinical variables in a group of 462 patients with Alzheimer's disease, 172 reported to have at least one first-degree relative with dementia and 290 classified with no family history. Patients with a presumptive family history differed from those without a family history in two ways: the course of dementia was described as having a fast rather than a slow progression from onset of symptoms to diagnosis, and caregivers reported a higher prevalence of family history of psychiatric disorders. There were no significant differences in age at onset, duration, female gender, aphasia and apraxia, handedness, family history of Down's syndrome, or number of children, brothers, and sisters. The association of faster course and family history of psychiatric disorders in the patients with a family history of dementia is consistent with the hypothesis of heterogeneity, but the overall results could also be explained by a genetic-environmental model of Alzheimer's disease.


The quality of life in the year before death.

Most Americans wish to live a long healthy life, but fear disease and dependency in their last years. Until recently, little has been known about the prevalence of opposite extremes of health in old age, particularly in the period leading up to death. We used results from the 1986 National Mortality Follow-back Survey to estimate proportions of elderly decedents who were "fully functional" or "severely restricted" in the last year of life. Estimates were based on responses from proxies to questions regarding the decedent's functional status, mental awareness, and time spent in institutions. Approximately 14% of all decedents aged 65 years and older were defined as fully functional in the last year of life; 10% were defined as severely restricted. Proportions varied with the decedent's age and sex, the underlying cause of death, and the presence of other preexisting conditions. Results from this survey and future surveys can be used to learn more about "successful agers"--their medical histories, their life-styles, and whether their relative number is increasing or decreasing overtime.


Failure of intragastrically administered Yersinia pestis capsular antigen to protect mice against challenge with virulent plague: suppression of fraction 1-specific antibody response.

We evaluated the Yersinia pestis capsular (fraction 1 [F1]) antigen as a potential oral immunogen in mice. We found that single doses of as much as 0.4 mg of F1, administered by intragastric (ig) intubation, were unprotective and did not stimulate production of detectable levels of specific antibody. Three weekly ig doses resulted in low serum antibody levels that also did not provide protection against challenge with virulent Y. pestis. Assays of type-specific antibody following intubation and subsequent challenge with a subcutaneous inoculation of F1 revealed that the quantity of antigen intubated and the secondary IgG2a antibody levels were inversely related, suggesting the induction of tolerance to intragastrically administered F1 antigen. Transfer of spleen cells from intubated mice to F1 immune recipients failed to demonstrate suppression of specific antibody, indicating that the immune tolerance observed in intubated mice was not due to a T suppressor cell-mediated effect. The results of this study indicate that Y. pestis F1 antigen is not likely to be an efficacious immunogen for oral vaccination of mice against plague.


Adjunctive intraoperative linear extrusion (Fogarty-Chin) balloon angioplasty.

Eighty-nine patients with 94 stenotic segments (mostly iliac or femoral) underwent balloon angioplasty with the first-generation (no guidewire) linear extrusion (Fogarty-Chin) system, in an adjunctive mode, and the overall long-term patency rate (mean follow-up: 21 months) was 81%. Patients were grouped into those having iliac or superficial femoral artery (SFA) lesions and subdivided according to the length of lesions. The overall primary and late success rates for iliac lesions were 95% and 86%, respectively, and for SFA lesions 91% and 76%, respectively. The primary and late success rates for iliac lesions less than 2 cm were 100% and 96%, respectively, and for iliac lesions 2 cm to less than 5 cm 92% and 80%, respectively. The primary and late success rates for SFA lesions less than 2 cm were 100% and 100%, respectively; for lesions 2 cm to less than 5 cm 100% and 83%, respectively; and for lesions 5 to 10 cm 83% and 67%, respectively. A stratified analysis by vessel and segment length reveals that, in SFA lesions with a segment length greater than 5 cm, there is a significantly lower patency rate (67%) when compared with the combined results of the Fogarty-Chin balloon angioplasty system in iliac and femoral artery lesions less than 5 cm (92%). In comparing the composite results presented in a recent text on endovascular surgery by Moore and Ahn as the base data for the standard coaxial (Gruntzig) balloon system, our results (short and long term) are similar.