Oral bioavailability of griseofulvin from aged griseofulvin:lipid coprecipitates: in vivo studies in rats. The bioavailabilities of aged coprecipitates of griseofulvin (Gris), dimyristoylphosphatidylcholine (DMPC), or egg phosphatidylcholine (EPC) and cholesterol (CHOL) in rats and correlations with their in vitro dissolution behaviors were determined. In vivo absorption studies of Gris:DMPC (4:1, w/w) or Gris:DMPC:CHOL [4:1(1:0.33 mole ratio)] coprecipitates yielded evidence of a 40% increase in the peak concentration in plasma (Cmax) and a 2.5- to 3-fold decrease in the time to reach Cmax (tmax), compared with those obtained with micronized Gris but a statistically unchanged area under the concentration in plasma--time curve (AUC) when 1-day-aged samples at equivalent doses were used. On the other hand, a 10% decrease in the AUC, a 20% increase in the Cmax, and a three- to fourfold decrease in the tmax were observed for the same formulations aged for 90 days. In comparison, the Cmax produced by the 1-day-aged Gris:EPC:CHOL [4:1(1:0.33 mole ratio)] coprecipitate was the same as that produced by micronized Gris, but the tmax and the AUC were significantly lower; the Cmax produced by the 90-day-aged coprecipitate was 30% higher than that produced by micronized Gris, but the tmax and the AUC remained unchanged. The Gris concentrations after 5 and 30 min (dissolution parameters) and the percent dissolution efficiency also showed excellent correlations with the concentration in plasma after 1 h, the Cmax, and the AUC (in vivo parameters) for all formulations, but the individual in vitro parameters showed poor correlations with the AUC results.(ABSTRACT TRUNCATED AT 250 WORDS)


Gastric emptying of tablets and granules in humans, dogs, pigs, and stomach-emptying-controlled rabbits. Rates of gastric emptying of nondigestible tablets and granules in humans were compared with those in three animal models: dogs, minipigs, and stomach-emptying-controlled rabbits. The rates of gastric emptying of both dosage forms in dogs tended to be faster than or similar to those in humans, whereas the rates in pigs were slower. In stomach-emptying-controlled rabbits, no tablets were emptied from the stomach because of their large size. The rate of gastric emptying of granules in rabbits was slow and variable. Food delayed gastric emptying in dogs, especially for tablets. In rabbits, the rate of gastric emptying of granules was faster when the granules were given before feeding, in comparison with that after feeding or under fasting conditions. We concluded that the dog is a better animal model for bioavailability studies under fasting conditions than the pig and the rabbit.


Simultaneous determination of otilonium bromide and diazepam by first-derivative spectroscopy. A rapid, simple assay procedure was developed for simultaneous analysis of otilonium bromide, a smooth-muscle relaxant, and diazepam in tablets containing 20 mg of otilonium bromide and 2 mg of diazepam (20:2 tablets) or 40 mg of otilonium bromide and 2 mg of diazepam (40:2 tablets) by "zero-crossing" first-derivative spectroscopy. The tablets were dissolved in 0.01 N HCl, mixtures were centrifuged at 3500 rpm (2472 x g) for 5 min, and first-derivative spectra were recorded. The absolute values of the derivative were measured at 264 nm for determination of otilonium bromide and between 406 and 408 nm (380 nm for analysis of 40:2 tablets) for determination of diazepam. The method is linear, quantitative, and reproducible and can also be used for the tablet dissolution test. Ten tablets of the same batch were analyzed by the described method and by a high-performance liquid chromatographic method, and the results were in good agreement.


Effects of verapamil, ouabain, and ethacrynic acid on calcium retention in perfused rat kidneys. A Sprague-Dawley rat kidney perfusion technique was used in situ to study the effect of verapamil, ouabain, and ethacrynic acid on renal calcium retention. The technique involves perfusion of the kidneys via the abdominal aorta and then through the left and right renal arteries and dorsal aorta. Verapamil (1 mM) in Krebs-improved Ringer solution increased calcium retention in the kidneys by approximately 117% compared with controls. With Na-free Krebs-improved Ringer solution, calcium retention increased by only 92%. However, in Krebs-improved Ringer solutions containing 59 and 122 mequivalents of Na, calcium retention in the kidney increased by 46 and 43%, respectively, compared with controls. With Krebs-improved Ringer solution containing 15 mM ouabain, calcium retention in the kidney decreased by 29.5%, whereas with 15 mM ouabain plus 1 mM ethacrynic acid in the perfusate, the effect on calcium retention in the kidney was insignificant compared with controls. These results suggest that two sodium-dependent calcium-transporting systems exist at the peritubular side of the kidney tubules: (1) a Na(+)-Ca+2 countertransport system sensitive to verapamil and (2) a Na(+)-Ca+2 cotransport system sensitive to the intracellular concentration of sodium.


Synthesis and anticholinesterase activity of new bispyridinium compounds. Synthesis of new bis(1-methylpyridinium) compounds containing a 1,4-diacetylbenzene linkage between the pyridinium moieties from commercially available 2-, 3-, and 4-picoline precursors was accomplished via metallation, reaction of the picolyllithium with 1,4-dicyanobenzene, and subsequent quaternization of the resulting bispyridyl compounds. Acetylcholinesterase inhibitory activity was determined colorimetrically with purified electric eel enzyme. Examination of structure-activity relationships indicated that the 3-substituted pyridinium compound is the most potent isomer, followed by the 2-substituted isomer, and that the 4-substituted analogue is the least active.


Structural properties of polyethylene glycol-polysorbate 80 mixture, a solid dispersion vehicle. The structural properties of the mixtures of polysorbate 80 with various polyethylene glycols (PEG), viz., PEG 1000, PEG 1450, PEG 3350, and PEG 8000, have been investigated by powder X-ray diffraction (XRD) and differential scanning calorimetric studies. These mixtures may be used as solid dispersion vehicles to insure complete dissolution of poorly water-soluble drugs. Although polysorbate 80 is a liquid at room temperature, the PEG-polysorbate 80 mixtures with up to 75% (w/w) polysorbate 80 were solid. The XRD studies revealed that the crystal structures (d-spacings) of the PEGs (M(r) 1000, 1450, 3350, and 8000) did not change with increasing amounts of polysorbate 80 in the mixture. The intensities of the XRD peaks, however, varied approximately in proportion to the concentration of PEG present. Similarly, the differential scanning calorimetric studies showed that the melting behavior of a PEG-polysorbate 80 mixture was similar to that of the PEG used. The lowering of the mp of a particular PEG due to the presence of 50% (w/w) polysorbate 80 in the mixture was < 6 degrees C, and the decrease in mp was < 12 degrees C in the presence of 75% (w/w) polysorbate 80. When enthalpies of fusion of the mixtures were normalized for the amounts of PEGs present, they were similar to those of pure PEGs. These results indicate that the crystalline structure of PEG in a PEG-polysorbate 80 mixture is substantially the same as that of the pure PEG, and that polysorbate 80 is incorporated into the amorphous region of PEG solid structure.


Effect of geometric factors on hydration kinetics of theophylline anhydrate tablets. The effect of tablet thickness on the hydration kinetics of types I and II theophylline anhydrate tablets at 95% relative humidity and 35 degrees C was studied with various kinetic equations. Samples of 1- and 2-cm-diameter tablets (1 g) were compressed at 1000 kg/cm2. Types I and II tablets expanded by 11-17% in volume during hydration to the monohydrate. The thickness expansion of all tablets exceeded the diameter expansion. After the hygroscopicity test, the final expansion ratio of type I tablets was more than that of type II tablets. The hydration kinetics analysis suggested that the hydration of theophylline anhydrate tablets proceeds as follows: Hydration of the 1-cm-diameter type I tablet followed the two-dimensional phase boundary equation, and that of the 1-cm-diameter type II tablet followed the three-dimensional phase boundary equation, but those of the 2-cm-diameter types I and II tablets followed the equation for two-dimensional growth of nuclei. The hydration kinetics of tablets does not depend on the nature of bulk powder but on geometrical factors (tablet thickness and porosity).


Physical and lubrication properties of magnesium stearate. The lubrication properties of two commercial-grade magnesium stearates were studied. Their moisture contents and crystal structures were similar. There were minor differences in their fatty acid composition, but the differences did not affect the lubrication properties. The lubrication properties correlated with particle size distributions and specific surface area. The effect of these parameters was further studied with unmilled and milled chemically pure magnesium stearate. Milling decreased the particle size and increased the specific surface area. In both cases, the batch with a smaller particle size and larger specific surface area had considerably better lubricity.


Decomposition of alpha-hydroxyaryl ketones and characterization of some unusual products. Alpha-Hydroxyaryl ketones such as 2-hydroxypropiophenone and 1-(2,4-difluorophenyl)-2-hydroxy-1-propanone, the key intermediates in the preparation of antifungal agents, decompose into oxidized, rearranged, and condensed products. These products were isolated and characterized. The possible mechanisms for the formation of the products are discussed.


A thin-layer chromatographic method to determine process impurities in leucovorin calcium. A densitometric thin-layer chromatographic method for the analysis of process impurities in leucovorin calcium was developed and validated. Using this method, folic acid, N10-formyldihydrofolic acid, p-aminobenzoic acid, and N-(4-aminobenzoyl)-L-glutamic acid were monitored with a limit of detection of approximately 0.1% each. Both absorbance and fluorescence densitometric evaluations were utilized; fluorescence evaluation selectively detected impurities that were not chromatographically resolved from leucovorin.


Pharmacokinetic characterization and tissue distribution of the new glucocorticoid soft drug loteprednol etabonate in rats and dogs. Loteprednol etabonate, a new glucocorticoid soft drug with a characteristic chloromethyl ester function in the 17 beta-position, is currently in the early phases of clinical development. As the basis for human trials, this study describes a new reversed-phase high-performance liquid chromatographic method for the determination of levels of drug in plasma and urine samples and assesses the pharmacokinetic properties of loteprednol etabonate in dogs and rats. Intravenous administration of loteprednol etabonate (5 mg/kg) to dogs revealed a terminal half-life of 2.8 h, a volume of distribution of 3.7 L/kg, and a total body clearance of 0.9 L/h/kg. Intact loteprednol etabonate was not detectable in the urine. After oral administration of the drug (5 mg/kg) to dogs, only metabolites, but no intact drug, were found in the plasma, an indication for a high first-pass effect. A pronounced binding of the drug to plasma protein (> 90%) and a high erythrocyte-buffer partition coefficient of 7.8 were determined in vitro. Preliminary information about tissue distribution and possible metabolic pathways were obtained in rats after oral administration of a 14C-labeled loteprednol etabonate suspension (5 mg/kg). pH-selective extraction into ethyl acetate revealed three distinguishable fractions: (1) a neutral lipophilic fraction, presumably intact drug, (2) an acidic, lipophilic fraction, and (3) a hydrophilic nonextractable fraction. Levels of intact drug and metabolites were highest in liver and kidney, whereas significantly lower levels were found in other investigated organs (lung, brain, heart).(ABSTRACT TRUNCATED AT 250 WORDS)


Liquid chromatographic determination of oxytetracycline in bovine plasma by double-phase extraction. A liquid chromatographic method for the determination of oxytetracycline in bovine plasma was developed. Contrary to most current tetracycline assays involving solid-phase extraction, oxytetracycline and the internal standard tetracycline were isolated from buffered plasma by double-phase extraction with ethyl acetate-isopropyl alcohol. The latter component was found to be essential to ensure reproducible partitioning of both tetracyclines into the organic layer. Chromatography was conducted on a Lichrosorb RP8 column, compounds were eluted with 0.01 M oxalic acid-methanol-acetonitrile, and detection was at 357 nm. Linearity was observed over the 0-8.5-micrograms/mL range, and the detection limit approximated 5 ng/mL. The extraction yield was 80.4 +/- 3.4% (n = 12), and the coefficients of variation for repetitive within-run and between-run analyses at two concentration levels (0.25 and 2 micrograms/mL) were 1.0-4.3 and 2.4-7.2%, respectively. The method was applied to a pharmacokinetic study of oxytetracycline in three nonlactating cows after a single intramuscular administration of a long-acting preparation.


The synthesis and anti-inflammatory properties of a new sulindac analogue synthesized from natural safrole. The synthesis of the new sulindac (3) analogue (Z)-5,6-methylenedioxy-2-methyl-1-(p-methylsulfinylbenzylidene)-3- indenyl acetic acid (1) from natural safrole (2), an abundant natural product occurring as the principal chemical constituent of Sassafraz oil, is described. The principal feature of this route is shortness, stereoselectivity, and high overall yield. The new analogue is produced in a yield of approximately 30% from the natural product. The results include the anti-inflammatory activity of 1 as well as that for its corresponding sulfide (12), a synthetic precursor of 1 that may be an important metabolic product of 1 by analogy to 3 itself. The anti-inflammatory profiles of these derivatives, measured in the carrageenan-induced rat paw edema test, indicated a 50% effective dose of 42 mg/kg for 1 and of 23 mg/kg for 12, confirming the structure-activity relationships of these agents. These results indicate that the new analogue 1 could represent a prodrug with a similar therapeutic profile to that of the pharmaceutical product 3.


Pharmacokinetics of moxisylyte in healthy volunteers after intravenous and oral administration. The concentration-time profiles of metabolites of moxisylyte, an alpha-blocking agent, in the plasma and urine of 12 healthy volunteers were investigated after intravenous (iv) and oral (two formulations) administration. The study was conducted with an open, randomized Latin squares design. Plasma and urine levels of moxisylyte and its biotransformation products were assayed by a specific HPLC method with fluorescence detection. Plasma levels declined in a monophasic or biphasic pattern depending on the subject. Two metabolites, conjugated desacetylmoxisylyte (DAM) and conjugated monodesmethylated DAM (MDAM), were found in plasma and urine. Unconjugated DAM was found in plasma only after iv administration. The apparent elimination half-lives of unconjugated DAM, conjugated DAM, and MDAM were 0.86, 1.7, and 3 h, respectively. The total amounts of metabolites (expressed as the equivalent of DAM) excreted in the urine were 75% after i.v. administration and 68 and 69% after oral administration of the two formulations. Oral absorption appeared to be complete for the two treatments. There was no statistical difference between the two oral formulations studied.


Genes, viruses and neurodevelopmental schizophrenia. Recent neuroimaging and neuropathological studies suggest a developmental origin for schizophrenia. Some cases may, therefore, be caused by a genetic defect in the specification of brain development. Early environmental hazards such as obstetric complications, and maternal exposure during pregnancy to influenza epidemics, have also been found to increase the risk of later schizophrenia. The relationship between the prevalence of influenza and birth date has been found more consistently for female than male schizophrenics. Female schizophrenia is also associated with a higher risk of schizophrenia in first degree relatives. This raises the question of whether part of the genetic predisposition to schizophrenia may comprise an abnormal reaction to maternal influenza.


The concept of schizophrenia: the conflict between nosological and symptomatological aspects. A brief overview is given of the history of psychiatric nosology during the last 150 years. The varying role of genetics for theories of nosology is described. Classifications of mental disorders, more or less based on nosological concepts, are mentioned, as well as the recent emphasis on operational tools for classification. Conflicts between nosology and modern classifications are mentioned. The place of the schizophrenic syndrome in relation to nosological theories is described, including the questions of schizophrenia being a disease entity, or several entities, or even a part of a continuum, or just a symptomatically characterized reaction form.


Genetic transmission and improved diagnosis of schizophrenia from pedigrees of adoptees. In previous investigations of the prevalence of schizophrenic illness among the biological relatives of schizophrenic adoptees in Copenhagen and the remainder of Denmark, the operation of heritable spectrum illness was clearly implicated. The findings supporting that conclusion are briefly summarized. Classical chronic schizophrenia was found almost exclusively in the biological relatives of chronic schizophrenic probands and its prevalence was ten times greater than that in the biological relatives of controls. These were global diagnoses, made without knowledge of the relationships and family histories of the subjects, and based upon the descriptions of dementia praecox or schizophrenia by Kraepelin and Bleuler. They showed considerably greater sensitivity and at least equal specificity in comparison with diagnoses made on the same material in accordance with operational criteria as exemplified by DSM-III. The prevalence of a disorder in the biological relatives of adoptees with that disorder in comparison with biological relatives of control adoptees offers a useful test for the expression of genetic factors in the disorder, but also a much needed evaluation of the validity of diagnoses based on clinical observation.


The importance of measures of affective temperaments in genetic studies of mood disorders. Collaboration between the University of Pisa, Italy, and the University of Tennessee, Memphis, U.S.A., on patients presenting with major depressive episodes (in the absence of nonaffective psychiatric illness) focused on the detection of depressive and hyperthymic temperaments. From our data on symptomatology, family history and course of 538 such patients, several findings emerge of cardinal relevance to genetic studies. Hyperthymic temperament, observed more commonly in men, appears as one pole of an attenuated form of manic-depressive illness. Thus, major depressives with this temperament have high rates of bipolar family history, even in the absence of hypomanic and manic episodes. The depressive temperament, more prevalent in women, is correlated with earlier onset and higher number of depressive episodes, greater severity of the Hamilton Rating Scale for Depression (HAM-D), as well as higher familial loading for mood disorders, compared with major depressives without this temperament. Building on Akiskal's latest model on the multifactorial origin of mood disorders, we submit that these temperamental dysregulations constitute the intermediate step between predisposing familial-genetic factors in affective illness and gender-related clinical expressions of mood disorders. The authors recommend that future high-risk prospective studies and genetic investigations should include measures of affective temperament.


Genetics of affective disorders.

Genetics of affective disorders. Studies of the mode of inheritance of affective disorders have been reviewed. The overall picture which emerges does not seem to involve specific modes of inheritance. Linkage studies based on the single gene hypothesis, support X-linkage for a bipolar variant, as well as possible linkage to chromosomes 6 and 11. The more recent approach of linkage studies using large numbers of RFLP markers scattered throughout the genome looks promising, though it may still give rise to misinformation. Several confounding factors have been suggested such as genetic and phenotypic heterogeneity, as well as the lack of well defined diagnostic criteria.


The complexity of psychiatric nosography and the "simplicity" of molecular genetics. Are molecular genetic approaches, related at the present time to psychiatric categories, mostly conventional? If the psychiatric field is to be related with the plasticity of the brain, and then to its ability to be reorganized relative to experiences, it seems to be more important to study the relation between genetics and brain plasticity. It therefore seems more significant today, to be raising questions rather than gathering data.


Tracking disease genes by reverse genetics. Increasingly, human genes are being identified by the "reverse genetics", or "positional cloning" approach. This molecular genetic strategy is particularly useful in mental illness, for which no readily detectable functional alterations are present to indicate candidate genes. The positional cloning procedure is briefly described. Significant examples of successful positional cloning are presented, including the fragile-X mental retardation syndrome gene. The study of gene expression may be complicated by genetic and non-genetic variability. Genomic imprinting may play a role in several mental illnesses, and may provide an explanation for the unusual inheritance pattern in fragile-X syndrome, for the phenotypic differences observed between Angelman and Prader-Willi syndromes, and for the juvenile onset form of Huntington disease. DNA instability may explain disease anticipation in fragile-X syndrome and myotonic dystrophy. Finally, the prospects of improvements in positional cloning methods for tracking genes responsible for mental illness are briefly discussed.


How genetically heterogeneous are the major psychiatric disorders? Linking specific chromosomal regions with the abnormal genes responsible for major psychiatric disorders is rendered particularly difficult because of three factors: diagnostic uncertainty; the possibility that some psychiatric disorders are caused by the concerted action of more than one gene; and the possibility that similar disorders may result from alternative genetic abnormalities. The approach proposed here is to concentrate initially on the highly focused problem of resolving the question of genetic heterogeneity of a given disorder. To this end, it is recommended (a) that studies concentrate on pedigrees large enough to contain at least two nuclear families with several affected individuals, and (b) that, within them, work be confined to an intensive examination of such small nuclear families containing an affected parent and at least two affected offspring with a fairly unmistakable phenotype--bipolar disorder, type I. By using molecular and genetic techniques designed to extract maximal information from this relatively small number of affected individuals, it should be possible to determine whether or not bipolar disorder is genetically heterogeneous. This is a limited but very important question, and the technology is presently available to answer it. The answer should, in turn, guide subsequent approaches to identification of the relevant gene or genes.


Extending the phenotype of schizophrenia: implications for linkage analysis. On the basis of simulation studies, we suggest that most existing designs for studying linkage in schizophrenia do not have sufficient power to detect a major contributing locus, even if one is present. For this reason, recent failures to replicate reports of linkage in schizophrenia are not surprising. Inclusion in the linkage design of phenotypes genetically related to schizophrenia, but more commonly found in the relatives of schizophrenics than schizophrenia itself, may increase the power of linkage analysis substantially.


From an animal model of an attentional deficit towards new insights into the pathophysiology of schizophrenia. The paper presents an animal model of schizophrenic-like attentional deficit, consisting of an inability to ignore irrelevant stimuli. It is based on the paradigm of latent inhibition (LI), in which animals learn to ignore repeatedly presented stimuli not followed by meaningful consequences. In a series of experiments it was demonstrated that the capacity to ignore irrelevant stimuli is lost in rats treated with systemic or intra-accumbens injections of amphetamine, in normal volunteers given amphetamine, in high "psychosis-prone" persons, in acute schizophrenic patients and in untreated male adult rats that were raised until weaning under conditions of extremely restricted stimulation. In addition, LI is lost following the disruption of the hippocampal input to the nucleus accumbens. In all of the above conditions tested for antagonism by anti-psychotic drugs a loss of LI is reversed. On the basis of these results we propose an animal model which accommodates a neurodevelopmental dysfunction, hippocampal pathology, mesolimbic DA overactivity, vulnerability to stress, and gender differences, all of which have been postulated as factors in the pathophysiology of schizophrenia.


On familial factors in the attentional deficit in schizophrenia: a review and report of two new subject samples. We review the literature on attentional impairment in schizophrenics and their first-degree relatives and present new information from ongoing family studies of the disorder in Ireland and Israel. Subjects were administered a neuropsychological test battery (the NIMH Attention Battery) intended to measure four different elements of attention: encode, focus/execute, sustain, and shift. Results from both samples indicated that schizophrenic subjects performed most poorly on the tests, the control subjects performed best, and the scores of the relatives fell somewhere between the other two groups. Separation among subject groups was most significant for those tests comprising the focus/execute and sustain elements. Some of the results indicate, as well, that tests of auditory sustained attention may be the most discriminating of all. Overall comparison of the Irish and Israeli cohorts revealed striking differences. While within cohort differences remained, subjects from the Irish sample performed more poorly on many of the tests than Israeli subjects. This between-cohort difference was not found in the sustain element, as measured by the Continuous Performance Test. The socio-cultural implications of these findings are discussed with respect to future neuropsychological studies of schizophrenia.


A summary of attentional findings in the New York High-Risk Project. We summarize here our findings with respect to attentional impairment among offspring of schizophrenic, affectively ill, and normal parents followed from childhood to adulthood during the two decades of the New York High-Risk Project (NYHRP). We review our data first, on childhood attentional performance in each of our two independent samples of such subjects and, second, on the relationship of early dysfunctions observed in this domain to psychopathological outcomes in adolescence and adulthood. Our cumulative results contribute strong support to the contention that global attentional dysfunctions may be viewed as a biobehavioral marker for the genetic liability to schizophrenic disorders.


Behavioral markers of schizophrenia useful for genetic studies. The advent of powerful molecular biological techniques have already led to the discovery of chromosomal loci linked to some genetically transmitted diseases. These techniques, however, lose their power if applied to a disease trait that is not Mendelian in its transmission. The low familial prevalence of psychiatric diseases such as schizophrenia make these techniques unsuitable for linkage studies of these conditions, if identification of schizophrenia relies solely on the clinical manifestation of the schizophrenic psychosis. Broadening the disease phenotype in diseases such as schizophrenia, with low recurrence risk, and narrowing it in diseases such as major affective disorder, with very high recurrence risk, may be an effective strategy for linkage studies of these diseases. Several alternative phenotypes are discussed, including smooth pursuit eye movement abnormalities, event related potentials, and deficient attentional deployment as measured by the continuous performance test. The strategy assumes that schizophrenia is a pleiotropic disorder, and that the psychosis is the rare form of the condition. The paper focuses principally on smooth pursuit eye movement abnormalities, and claims a plausible place for them as an independent expression of schizophrenia. With this strategy, the possibility for successful linkage studies increases, since familial distributions of schizophrenia and pursuit abnormalities, considered together, appear to fit an autosomal dominant pattern.


Gene expression in mental illness: a navigation chart to future progress. An initial course in disentangling complex causal interactions in psychiatric illnesses, we suggest, is finding co-familial traits with classical Mendelian segregation. Starting with non-Mendelian traits, three methods can be used to find underlying Mendelian phenotypes. (1) Statistically-inferred latent traits, with more nearly Mendelian transmission than the measures from which they are derived, can serve as pointers to concrete Mendelian phenotypes. (2) Linkage of non-Mendelian traits to genetic markers, if it can be established, can be followed by searching for phenotypes that discriminate carriers from non-carriers of the imputed trait gene. (3) In the long run, the most successful method is likely to be direct refinement of non-Mendelian behavioral and physiological traits into more fundamental components.


Subtypes of depression, efficacy, and the Depressive Experiences Questionnaire. The purpose of our study was to reanalyze the Depressive Experiences Questionnaire (DEQ; Blatt, D'Afflitti, & Quinlan, 1976, 1979) and to evaluate relationships between the questionnaire's factors and other construct-related measures, the Beck Depression Inventory (Beck, Ward, Mendelson, Mock, & Erbaugh, 1961) and the Edwards Personal Preference Schedule (Edwards, 1959). We argue that composite scores derived from this study's principal components analysis are more interpretively useful than the original factor scales reported by Blatt et al. (1976; Blatt, Quinlan, Chevron, McDonald, & Zuroff, 1982). Correlational evidence reported in this study does not support the Dependency and Self-Criticism factors as adequate representations of anaclitic and introjective depression (Blatt, 1974). The third DEQ factor scale, Efficacy, as revised for this study, demonstrated moderate to strong associations with other personality measures.


Effects of stimulus color, pattern, and practice on sex differences in mental rotations task performance. Redundant color information improved performance for both sexes on the Shepard Mental Rotations Task (MRT; Shepard & Metzler, 1971). Absolute score gains for women were larger than those for men; therefore, relative improvement was greater. Substantial practice effects, also favoring women, were apparent in both studies. Study 1 showed that redundant color improved performance by 0.25 SD. Study 2 demonstrated that redundant black-and-white pattern information did not have any effect; a second visuospatial channel, redundant color, was a critical factor in improving scores of men and women on difficult mental rotations tasks.


Unhappy self-esteem: a meaningful exception to the rule. Self-esteem and well-being are often seen as closely related and sometimes even considered conceptually identical. In marked contrast to this association, the focus of this article is on experiences of a high level of self-esteem coexisting with a low level of well-being: unhappy self-esteem. Although these experiences constitute an exception to the general rule, they nevertheless have a theoretically coherent character. In this article, a framework was used in which the self was considered an organized process, and an idiographic assessment procedure--the method of self-confrontation--was called upon. The self-statements of 15 subjects in the Netherlands showing unhappy self-esteem were compared, and the phenomenon was found to clearly refer to anger and opposition.


The ability to encode facial and emotional expressions by educable mentally retarded and nonretarded children. The object of the present study was to investigate whether 20 educable mentally retarded (EMR) children matched for verbal mental age on the Peabody Picture Vocabulary Test (PPVT; Dunn, 1965) with 20 nonretarded (NR) controls were adept in identifying emotional facial expressions and producing the equivalent word adjectives. I also reassessed the relationship between the phase of identifying facial expressions (i.e., happy, sad, angry, and scared) and the phase of producing emotional word adjectives, consisting of 16 emotional linguistic constructions (4 short stories for each of the adjectives) between higher in verbal ability educable mentally retarded (HEMR) children and higher in verbal ability nonretarded controls (HNRC) and between low in verbal ability educable mentally retarded (LEMR) children and low in verbal ability nonretarded controls (LNRC). There were no significant differences between EMR and NR children in general, despite the fact that EMR children had deficits in receptive linguistic competence.


Treatment of persistent and complicated pancreatic pseudocysts. In a review of 1895 patients admitted with pancreatitis during a 4-year period, 241 (12.7%) were identified as having pseudocysts. The majority of these were treated without operation, but 59 patients (24.5%) needed surgical intervention because of persistence (17 cases) or development of complications (biliary obstruction in 16, infection in 12, duodenal obstruction in ten and haemorrhage in four). Most cysts (68%) resulted from alcohol-related chronic pancreatitis. Blunt abdominal trauma was the cause in three. Operations included internal drainage in 35 (cystogastrostomy in 23, cystojejunostomy with Roux-en-Y in ten and cystoduodenostomy in two), external drainage in 20, pancreatic resection in two, and gastroenteric or bilioenteric bypass in ten. There were six postoperative deaths (10.2%), one after internal drainage (3%) and 5 (25%) after external drainage (P < 0.01, Fisher's exact test). Pseudocyst decompression failed to relieve biliary obstruction in half of the patients and biliary-enteric anastomosis was necessary because of a stricture in the distal bile duct. Massive bleeding from pseudocyst-related false aneurysms was successfully controlled by transcatheter angiographic embolization in four patients. During 1-5 years' follow-up, 24 of the 53 surviving patients (45%) were readmitted with pancreatitis and three of these died. Pseudocysts recurred in three patients, with spontaneous resolution in two and need for operation in one. It is concluded that operative treatment of complicated pseudocysts carries a substantial mortality rate. The need for additional biliary-enteric bypass after cyst decompression should be carefully assessed during operation. Angiographic embolization of pseudocyst haemorrhage is a valuable therapeutic manoeuvre.


Therapeutic options in subclavian artery disease. Chronic ischaemia of the upper limb secondary to subclavian artery disease is uncommon. These patients may present with a variety of symptoms caused by cerebrovascular or upper limb hypoperfusion. If these symptoms are severe, revascularization may be required. This may be accomplished by percutaneous transluminal angioplasty or surgery. The experience of the management of these patients has been reviewed to document their presentation, treatment and outcome.


Retrospective analysis of the Bangour Burn Unit. The Bangour Burn Unit was opened in 1968 having been built using existing buildings modified in a relatively inexpensive way. The buildings were unsophisticated because the Unit was designed to be temporary before an eventual move into a new regional plastic surgery unit elsewhere. An analysis of some salient observations over the 22-year period of its lifetime is discussed.


Ambulatory hydrocele surgery: a review of 50 cases. In a prospective study, 50 cases of hydrocele were treated by ambulatory surgery. Lord's technique was performed under local anaesthesia. A modified simple scrotal dressing was used to facilitate ambulation. In one case (2%), haematoma was reported. There were no wound infections. Patients tolerated the procedure well and ambulation was excellent in all cases. On the grounds of safety and cost effectiveness, most hydrocele repair procedures should be performed on ambulant patients.


Excessive wastage of blood resources in elective colorectal surgery. A prospective study of perioperative blood transfusion requirements for elective colorectal surgery over a 6-month period has been conducted. A total of 106 procedures was performed, 94 of which were for malignancy. A median of 2 units of packed red cells was cross-matched per patient. Twenty-three patients (21.7%) required a transfusion in the postoperative period. Fourteen of these patients required a second cross-match sample because their transfusion occurred more than 48 h after the original request. The greatest transfusion requirements were for patients undergoing abdominoperineal resection or subtotal colectomy. Routine perioperative cross-matching and reservation of blood for elective colorectal surgery may be an unnecessary expense, resulting in wastage of resources. A selective policy of serum being retained for grouping in these patients is justified.


Adverse influence of fibrin sealant on the healing of high-risk sutured colonic anastomoses. The effect of fibrin glue sealing on sutured colonic anastomoses was studied using a 'high-risk' colon anastomosis model in the rat. Animals (n = 104) were randomized to have their sutured anastomosis sealed with fibrin glue or left untreated. They were assessed clinically until they were killed on the fourth day after surgery when contrast radiology, detailed post-mortem examination, anastomotic bursting pressure (ABP) and assessment of adhesion formation were performed. The clinical outcome was worse in the glued group (toxic or death from sepsis: 18 versus seven in the non-glued group; P = 0.0354), which also showed a significantly higher moderate to major leak rate (17 versus two in the non-glued group; P = 0.0009). The median ABP was significantly higher in the glued anastomosis group (96 versus 68 cmH2O in the non-glued group; P = 0.0367). Excessive perianastomotic adhesion formation was significantly greater in the glued group. Microscopic examination showed an extremely intense inflammatory reaction in the glued anastomoses compared with that in the untreated group. These results indicate that sealing of a sutured anastomosis with fibrin glue containing an antiproteinase impairs healing the of anastomotic wound, probably by resisting the ingrowth of vascular granulation tissue during the early stages of repair.


Tissue expanders in the gut: a histological and angiographic study. A study to assess the effects of an intraluminally placed tissue expander in the rabbit jejunum was performed. Oval expanders (8 cm3) were placed via a midline coeliotomy through an enterotomy on the antimesenteric border of the jejunum. The reservoir was tunnelled subcutaneously through the abdominal wall. Expansion was carried out through this port at 7-10-day intervals using 0.5-cm3 increments. At the end of the experiment, angiography was performed and routine haematoxylin and eosin-stained sections were produced. Angiography revealed an increase in vessel diameter and vascular density of the specimen. Histology showed an increase in villi size, an increase in the number of goblet cells and a marked increase in the thickness of the muscular layer. The possible applications of this technique are discussed.


Wrist and hand ganglion treatment with hyaluronidase injection and fine needle aspiration: a tropical African perspective. In an attempt to provide an alternative non-invasive treatment to surgical excision of ganglion cysts of the hand, and as part of the departmental audit resulting from the prevailing economic depression, 340 consecutive patients with 349 ganglia were treated in a prospective investigation by intralesional injection of hyaluronidase (up to 150 units in 1 ml) followed by fine needle aspiration (FNA) of the cyst to dryness. Pressure was applied over a piece of gauze and maintained with a crepe bandage for 24 h. Of the 340 patients treated in this way, the vast majority (323 or 95.0%) were considered to be cured on clinical examination at 6-month follow-up; only 17 patients (5.0%) exhibited recurrence during this period and these were successfully treated by re-aspiration. To the knowledge of this author, this is the first report of the use of the enzyme hyaluronidase as an adjunct to FNA in the treatment of ganglion cysts of the hand. The results clearly show that this method of treatment is a safe, fast, well accepted and cost-effective alternative to surgical excision, which is relatively expensive and is known to be associated with certain complications, including hypertrophic scars and cheloids.


Sensory nerve damage during surgery on the hallux. Division of the medial branch of the superficial peroneal nerve during surgery on the hallux can lead to unpleasant and troublesome symptoms, including neuroma formation. This study aims to show the incidence and consequences of damage to this nerve and to describe an easily applied solution. A total of 75 feet (51 randomly selected patients) was examined, with a mean follow-up of 4 years. Evidence of nerve damage was seen in 45% on clinical examination but, interestingly, only 29% were aware of their symptoms when questioned beforehand. In 3% the symptoms were severe and disabling. In view of this unexpectedly high incidence, ten cadaveric feet were dissected with a reliable surface marking being described to avoid the nerve during surgery.


Sex hormones in postmenopausal HLA-identical rheumatoid arthritis discordant sibling pairs. Dehydroepiandrosterone sulfate (DHEAS), testosterone, androstenedione, 17-beta estradiol and sex hormone binding globulin have been assayed in 50 HLA-identical postmenopausal rheumatoid arthritis (RA) discordant sibling pairs. The only difference was for DHEAS, siblings with RA having a significantly lower level than their sisters. However, in 68 patients with RA, the level of DHEAS inversely correlated with disease duration, a radiographic grading score, the Health Assessment Questionnaire score, duration of morning stiffness, and a clinical score of disease activity and severity (the Spread/Severity index). These observations, taken with that of previous work on DHEAS, suggest that low levels may be a consequence of RA rather than predisposing to the disease. The role of sex hormones in RA will have to be approached in alternative ways.


Autoantibodies to lipocortin-1 are associated with impaired glucocorticoid responsiveness in rheumatoid arthritis. Autoantibodies to the antiinflammatory protein lipocortin-1 have been found in patients with rheumatoid arthritis (RA) receiving oral glucocorticoids. The highest antibody titers correlated with a requirement for high maintenance doses of steroid (> 7.5 mg/day prednisolone). Forty-two patients with RA were grouped according to high or low autoantibody titer. In 18 patients, peripheral blood leukocyte counts and phenotypic analysis were performed before and 4 h after a single intravenous (iv) dose of 100 mg hydrocortisone. The group with low titer antibody exhibited a normal poststeroid peripheral blood lymphopenia, but the response in the group with high antibody titer was considerably blunted. In a 2nd study, 24 patients received 3 separate doses of 1000 mg iv methylprednisolone. After 8 weeks the group with the high titer antibody had shown no improvements in clinical or laboratory variables observed in the group with low titer antibody. Thus, the presence of high titer antilipocortin-1 antibody is associated with impaired responses to glucocorticoid therapy both in terms of clinical efficacy and effects on the immune system. This could explain the relative glucocorticoid resistance reported in a proportion of patients with RA.


The effects of methotrexate on interleukin 1 in patients with rheumatoid arthritis. The effect of methotrexate (MTX) treatment on patients with rheumatoid arthritis on interleukin 1 (IL-1) was evaluated. Eight patients received a weekly MTX dose of 7.5 mg for 6 weeks; none received concomitant corticosteroids. Peripheral blood was obtained 1 day before and 1 day after their weekly MTX dose at Week 1 and 6, isolated monocytes stimulated ex vivo with 1 mg/ml zymosan for 18 h and IL-1 measured by bioassay. Serum and plasma IL-1 levels were measured by immunoassay. Four patients manifested a sharp decrease in IL-1 production 6 weeks after MTX administration associated with a decrease in the number of painful but not swollen joints. No effect on blood levels was observed. Our results suggest that MTX may affect IL-1 production and IL-1 mediated events in some patients.


Nitrogen mustard as induction therapy for rheumatoid arthritis: clinical and immunologic effects. We treated 5 patients diagnosed with rheumatoid arthritis (RA) with nitrogen mustard (HN2) and monitored clinical and immunologic variables. HN2, 0.3 mg/kg ideal body weight was given over 7 days. Disease activity and immune function were monitored during and after treatment. Duration of morning stiffness (p = 0.0044), joint count (p = 0.0140), and assessment of pain (p = 0.0264) and function (p = 0.0057) improved by Day 6. T lymphocytes (p = 0.0060), especially T memory cells (CD4CD29; p = 0.0017) fell dramatically. HN2 is effective for rapidly gaining control of active RA. This effect is T cell specific.


Erythroid abnormalities in rheumatoid arthritis: the role of erythropoietin. Erythroid alterations were studied in 136 patients with rheumatoid arthritis (RA). Anemia was present in 75 cases. A definite diagnosis was determined in 65. The most frequent anemia was that of chronic disease (ACD) (43 cases); 14 patients with ACD presented with moderate to severe anemia. Prevalence of deficiencies were also high (15 cases had iron deficiency anemia, IDA). Serum erythropoietin levels were different in patients with RA compared with a healthy control group (p < 0.00001). Serum erythropoietin was increased in ACD (49 +/- 28.8 U/l) with respect to both RA (38.6 +/- 12.7 U/l, p = 0.0036) and controls (18.2 +/- 7.6 U/l, p < 0.00001). Although hemoglobin (Hb) was similar in ACD and IDA, serum erythropoietin in ACD was lower than in IDA (p = 0.01). There was a negative relationship between Hb and serum erythropoietin in ACD (r = -0.42, p = 0.005). In conclusion, almost 50% of patients with RA have anemia and ACD is the most frequent. As serum erythropoietin in ACD is blunted, patients with moderate to severe ACD are possible candidates for erythropoietin treatment.


Digital subtraction radiography for the assessment of bone changes in rheumatoid arthritis. A digital subtraction radiography (DSR) method was developed to assess changes of bony erosions in the hands of patients with rheumatoid arthritis (RA). The method initially was validated in vitro using simulated lesions in cadaver hands. The sensitivity, specificity and accuracy in detecting simulated lesions were 84.6, 100, and 92.3%, respectively. This technique was then pilot tested clinically using 17 patients with RA and 5 controls with no arthritis. All subjects received the nonsteroidal antiinflammatory drug, flurbiprofen, during the 6-month study period. No progressive changes in bone were observed in the control group; however, 18% of the target joints studied in the group with RA showed significant erosive changes (positive or negative) which correlated with the clinical assessment. The results indicate that DSR may be useful in detecting small changes in erosions on hand/wrist radiographs of patients with RA.


Growth and characterization of fibroblasts obtained from bronchoalveolar lavage of patients with scleroderma. We describe the detection and the growth of fibroblasts with human smooth muscle cell differentiation features from bronchoalveolar lavage (BAL) fluid of 53% of patients with scleroderma, but not from healthy controls. The binding of alpha-actin, vimentin and desmin antibodies by scleroderma lung fibroblasts exceeded that of normal adult lung fibroblasts, indicating that scleroderma lung fibroblasts express some markers of human smooth muscle cell differentiation (myofibroblasts), which may account for differences in biological behavior. A mesenchymal cell phenotype was documented by mRNA analysis, showing high expression of collagen type I and fibronectin in these cells. Fibronectin is also released in significantly higher amounts by scleroderma alveolar macrophages than by macrophages from healthy donors.


Chloroquine therapy in psoriatic arthritis. Our aim was to examine the efficacy of chloroquine in psoriatic arthritis (PsA) and to assess whether chloroquine therapy exacerbated psoriasis. Thirty-two patients had been given chloroquine therapy while attending the University of Toronto Psoriatic Arthritis Clinic. Twenty-four patients continued therapy for at least 6 months, and 18, or 75%, demonstrated > 30% reduction in the actively inflamed joint count. Moreover, there was a significant decrease in the number of actively inflamed joints. A control group, consisting of 24 patients taking no remittive agents, seen during the same period of time, was identified. Only 14, or 58%, of these patients had a > 30% reduction in inflamed joint count over a 6-month period. This pattern was not significantly different (p = 0.19) from the chloroquine treated group. Of the 32 patients who had been given chloroquine, a total of 6 had an exacerbation of psoriasis, only 1 discontinued therapy. There was no case of exfoliative dermatitis. Six of the 24 control patients had an exacerbation of psoriasis. Our experience suggests that chloroquine may be an effective treatment in PsA, and that it does not exacerbate psoriasis. A prospective, randomized, double blind, controlled trial of antimalarial therapy in PsA is warranted.


The utility of the arthritis impact measurement scales for patients with psoriatic arthritis. The Arthritis Impact Measurement Scales (AIMS) consists of 9 scales that measure physical function, pain and psychosocial function. It has been validated for use in various forms of arthritis, but not in psoriatic arthritis (PsA). The AIMS was administered to 145 patients attending our PsA clinic. We carried out simultaneous assessment of clinical measures of function, measures of disease activity, and measures of disease severity. Most scales of physical function were moderately to highly correlated with clinical measures of function (r = 0.33-0.57; p = 0.0001), measures of disease activity (r = 0.24-0.53, p = 0.003-0.0001), and measures of disease severity (r = 0.23-0.6; p = 0.02-0.0001). The pain scale was highly correlated with clinical measures of function and measures of disease activity (r = 0.38-0.58; p = 0.0001) but not with measures of disease severity. Of the psychosocial scales, the depression scale was moderately correlated with clinical measures of function (r = 0.27-0.3; p = 0.001-0.0001). Our data suggest that the physical function and pain scales are good indicators of overall function and disease activity and are valid for use in PsA.


Autoimmune thyroid disease, thyroid antibodies and giant cell arteritis: the supposed correlation appears fortuitous. The coexistence of giant cell arteritis (GCA) and autoimmune thyroid diseases has been suggested. In our prospective study of 39 patients with GCA, we measured autoantibodies and thyroid function and compared the results to those of a control group. No statistical difference was found. Our conclusions differ from those of other authors.


A new protein antidenaturant agent, bindarit, reduces secondary phase of adjuvant arthritis in rats. Bindarit (or 2-[(1-benzyl-indazol-3-yl)methoxy]-2-methyl propionic acid) reduces heat induced denaturation of bovine and rat serum albumin in vitro (EC50 = 8.5 and 65 micrograms/ml, respectively) and inhibits heat induced serum albumin denaturation after in vivo (12.5-25-50 mg/kg po) administration in rats. To assess the relationship between protein denaturation and the development of chronic inflammatory diseases, the drug (0.5 or 0.12% medicated diet) was studied in comparison with indomethacin (1 mg/kg po daily) in rats injected with complete Freund's adjuvant. Bindarit appeared different from aspirin-like drugs, antiinflammatory steroids and immunosuppressants because it does not reduce primary inflammation of arthritic rats and was shown to be completely inactive on cyclo and lipooxygenase activity in vitro and on immune reactions of mice in vivo. Nevertheless, the drug strongly reduced the development of the secondary phase of adjuvant induced arthritis. The most significant effect of bindarit in this phase was a strong inhibition of serum albumin denaturation in arthritic rats. Assessment of both electrophoretic and quantitative changes suggests that the reduction of albumin during inflammation is due, at least in part, to a denaturation of native albumin, which loses its electrophoretic mobility. The involvement of protein denaturation in the production of new antigenic determinants, their pathogenic relevance in the development of adjuvant arthritis and the possibility that protein stabilization by bindarit could be the mechanism of action of the drug are discussed.


Epidemiology of Lyme disease in Olmsted County, MN, 1975-1990. Lyme disease has been reported in Minnesota since 1980. Until now, it has not been regarded as endemic in Olmsted County. We reviewed the medical records of all 68 cases of Lyme disease diagnosed in residents of Olmsted County for 1980-1990. Only 17 of the 68 cases met Centers for Disease Control surveillance criteria for definite Lyme disease (13 men, 4 women). Of these 17 cases, 3 were likely acquired in Olmsted County. The age and sex adjusted incidence rate was 2.3/100,000 (95% CI 1.1-3.4). Although reporting of Lyme disease is mandatory in Minnesota, only 7 suspected or definite cases were actually reported to public health officials. Of these, 4 met the case definition. In this population Lyme disease was both over-diagnosed and underreported. This observation questions current data and could have important health policy implications if confirmed nationally.


Measurement of synovial fluid volume: a new dilution method adapted to fluid permeation from the synovial cavity. A new dilution method was developed to measure the synovial fluid (SF) volume in a single knee joint of a rabbit. In this dilution method, we used 2 different kinds of dextrans, one having a mean molecular weight (M(r)) of 487,000 and labelled with fluorescein isothiocyanate (FITC) as a marker, and the other M(r) of 300,000 to give hypertonicity to the solution. By employing these high M(r) dextrans in their optimal concentrations, we were able to minimize the inevitable transsynovial shift of the fluid and marker after intraarticular injection. The original SF volume was then calculated using the fluorescence intensities of the injected and retrieved fluids. By determining the accurate volume of SF, it became possible to measure per joint the protein and hyaluronic acid contents in SF as well as the total and differential cell counts. The results obtained by this new method were reasonable and consistent, except for volume, with the reported results based on pooled samples from multiple joints. We believe that the method can contribute to the analysis of SF changes under various conditions in small laboratory animals.


Effect of calcitonin or the anabolic steroid Decadurabolin on serum beta 2 microglobulin in osteoporotic postmenopausal women. beta 2-Microglobulin (beta 2m) is a small polypeptide of 99 amino acids with a molecular weight of 11.800. It is found in serum, on the surface of almost all mammalian cells and is part of the surface antigen of the cell membrane of human lymphocytes. Moreover, the heavy chain of major histocompatibility complex class I molecules normally associates in the membrane with beta 2m. According to Canalis, et al this polypeptide is a bone derived growth factor. In our study, serum beta 2m, osteocalcin bone Gla-protein (BGP) and calcium phosphorous metabolism were evaluated in a group of 18 osteoporotic women. Nine were treated with an anabolic steroid, DecaDurabolin and 9 with salmon calcitonin (sCT) for 3 months. The same variables were evaluated in a control group (6 osteoporotic women) treated with oral Ca (1,500 mg/day for 3 months). A significant increase in serum beta 2m and BGP (p < 0.001) was observed after anabolic steroid while a significant decrease in serum beta 2m and BGP (p < 0.01) was observed after sCT. No significant variation for serum beta 2m and BGP was observed in the control group. No significant variation of calcium-phosphorous metabolism was observed in either the control group or the treated group. BGP variations suggest that anabolic steroid stimulates bone formation while a decrease in BGP observed after sCT suggests that this hormone depresses bone turnover via a block in osteoclast activity. Moreover it could be suggested that anabolic steroid and sCT affect beta 2m producing cells.


Nocturnal pain correlates with effusions in diseased hips. Thirty-five of 50 patients with different hip joint disease had sonographic evidence of joint effusion. Arthrocentesis confirmed effusions in 30 of these 35 patients. Thirty-two of the 35 patients had nocturnal pain. Both nocturnal pain and sonographic evidence of effusion decreased after aspiration (15 patients) and aspiration and injection of corticosteroids (15 patients). In a further group of 61 patients who subsequently had Charnley arthroplasties, 35 had positive sonograms before operation. Of these, 25 had effusions confirmed at operation, the remaining 10 having synovitis and capsule thickening. Again a correlation was found with nocturnal pain. The sensitivity of sonography in detecting hip joint effusion was 92% with a specificity of 70%. Nocturnal pain had a lower sensitivity, 85%, but higher specificity, 94%.


Impact of running on lumbar bone density: a 5-year longitudinal study. Our study was designed to examine associations of longterm physical impact (running) with changes in lumbar bone mineralization. Study subjects were a volunteer sample of 14 members of a running club now aged 55 to 77 years and 14 matched controls undergoing computerized scans of the first lumbar vertebra both at baseline and after 5 years. Separate analyses included data from 23 runners available over the entire 5-year period. A decrease in bone mineral density (BMD) over time was statistically significant in both runners and controls. Among runners bone loss was most pronounced in those decreasing their running habits substantially. At the 5-year mark, runners maintained greater BMD. A highly significant correlation was found between change in lumbar BMD and average time spent running (min/week) over the 5-year period. Furthermore, changes in BMD were positively correlated with changes in run min/week. We conclude that regular running appears to reduce age related bone loss both in women and men over 50 years of age. However, substantial decreases in physical weight bearing activity are associated with important bone loss in the lumbar spine.


Extraction of calcium containing crystals from synovial fluids and articular cartilage. A protocol for extracting minerals from biological tissues and fluids has been developed. Synovial fluid (SF) and articular cartilage samples were treated with papain and sodium hypochlorite to extract and concentrate calcium containing crystals. The method was validated by testing the ability to reextract synthetic calcium pyrophosphate dihydrate and calcium hydroxyapatite crystals added to SF in an unchanged form. The extracted crystals were analyzed by infrared spectroscopy and X-ray powder diffraction. The concentration of both calcium pyrophosphate dihydrate and hydroxyapatite crystals in SF was found to be higher than published figures.


Clinical manifestations of the antiphospholipid syndrome. We studied a large cohort of patients with systemic lupus erythematosus (SLE) to determine which manifestations associate with the antiphospholipids (aPL) and to ascertain when 2 or more such manifestations coexist, the association with aPL is stronger and the titers tend to be higher. We have confirmed that when aPL occur within SLE, they may account for some disease manifestations. We also described a similar syndrome occurring in the absence of a primary condition which we termed primary antiphospholipid syndrome (APS). Thus, we were able to construct preliminary criteria for the classification of APS as it occurs in SLE. With appropriate additions and exclusions to rule out SLE, these criteria could be applied to the classification of primary APS.


The nature of antiphospholipid antibodies. Despite the striking clinical manifestations associated with antiphospholipid antibodies (aPL) the role of these autoantibodies in disease and the nature of their true "inducing" and "target" antigens remain elusive. To address these issues, we investigated the immunogenic potential of phospholipid structures. To date, phospholipid immunogens have included hexagonal (II) forms of phosphatidylethanolamine and mixtures of apolipoprotein H (beta 2-glycoprotein I) with cardiolipin. Both hexagonal (II) phosphatidylethanolamine and the cardiolipin/apolipoprotein H mixture were capable of inducing aPL with lupus anticoagulant activity. Bilayer phosphatidylethanolamine and cardiolipin in the absence of apolipoprotein H were nonimmunogenic. Our data support our views that specific phospholipid structures are recognized by the immune system and that such structures serve as inducing and/or target antigens in the pathogenesis of aPL in vivo.


Idiopathic musculoskeletal pain syndromes in children. Between 1982 and 1990, 81 children with localized or diffuse musculoskeletal pain, for which no cause could be found were seen in a pediatric rheumatology clinic. Forty-one children had localized idiopathic pain and 40 had diffuse idiopathic pain. Twenty-four of the patients with localized idiopathic pain fulfilled criteria for definite reflex neurovascular dystrophy. Thirty-five patients with diffuse idiopathic pain fulfilled criteria for fibromyalgia. Four patients with localized idiopathic pain (10%) developed diffuse idiopathic pain during followup; four patients with diffuse idiopathic pain (10%) had a history of localized idiopathic pain and one patient had previously been diagnosed as having Tietze's syndrome. Recurrences or persistence of pain was very common. Many children had potentially important stressors including single parent families, histories of sexual abuse, and learning difficulties. Idiopathic musculoskeletal pain is a common cause of referral to a pediatric rheumatology clinic and is often associated with significant morbidity.


Major histocompatibility complex class II alleles in Kawasaki syndrome--lack of consistent correlation with disease or cardiac involvement. Recent refinements in molecular genetic typing have allowed the precise determination of the extensive polymorphism now recognized in class II major histocompatibility complex (MHC) genotypes. This could have important applications in Kawasaki syndrome (KS), where the relative contribution of genetic factors is now well known. Accordingly, 44 Caucasian, 13 Asian, and 5 American black patients, as well as 221 Caucasian controls were typed for HLA-DRB1, DRB3, DRB4, DQA1, DQB1, and DPB1 alleles by oligonucleotide probe hybridization of polymerase chain reaction amplified genomic DNA using probes and primers supplied by the 11th International Histocompatibility Testing Workshop. Among the 15 HLA-DRB1, 3 DRB3, 9 DQA1, 15 DQB1, and 19 DPB1 alleles examined, none were found to be significantly associated with KS, except for an increased frequency of HLA-DRB3*0301 in Houston Caucasian patients when compared to Houston Caucasian controls (38 vs 11%, pc = 0.012, RR = 5.0). Twelve patients developed coronary artery involvement of whom 7 had aneurysms and 5 had dilatation (8 Caucasians, 2 blacks, 2 Asians). No specific HLA class II allele was associated with this disease complication. Despite a regional association, our data fail to support a consistent role for MHC class II alleles in the pathogenesis of KS.


Chlamydial associated syndrome of arthritis and eye involvement in young children. The current classification of juvenile rheumatoid arthritis (JRA) consists of several distinct subsets. We describe 6 children (2 boys, 4 girls, mean age 3.7 years, range 2.0-4.9 years) with arthritis and eye involvement associated with infection with Chlamydia trachomatis. In some of the children, the clinical picture was similar to early onset pauciarticular JRA: onset within the first 4 years of life, predominance of girls, pauciarticular arthritis, subacute uveitis, and presence of antinuclear antibodies. Joint involvement was pauciarticular in 4 patients and polyarticular in 2. Two patients had clinical symptoms of Reiter's disease. Further investigations of this post chlamydial associated syndrome should be performed to establish appropriate diagnostic, therapeutic and prognostic measures.


Juvenile dermatomyositis and pregnancy: report and literature review. Reports of dermatomyositis (DM) and polymyositis complicating pregnancy are rare. Only nineteen cases have been published. We describe the first case in which juvenile DM appeared during pregnancy. An emergency cesarean section had to be performed due to fetal distress at 37 weeks of gestation. Patient and infant are doing well after 8 months of followup.


Bilateral renal artery occlusion in a patient with primary antiphospholipid antibody syndrome: thrombosis, vasculitis or both? A 43-year-old man presented with oliguria and hypertension. Renal arteriography showed bilateral renal artery occlusion. Circulating antiphospholipid antibodies were found together with a change in natural anticoagulant plasma levels. Immunofluorescence of examined vessels showed immune complex vasculitis. We discuss the pathogenetic mechanism leading to the development of this rare occlusive event.


Ten year survival of a patient with advanced pulmonary hypertension and mixed connective tissue disease treated with immunosuppressive therapy. Pulmonary hypertension is the most frequent cause of death in patients with mixed connective tissue disease. The pulmonary hypertension is progressive and difficult to treat effectively. These patients experience considerable morbidity secondary to diminished cardiac function. We describe a case in which sequential administration of cyclophosphamide and cyclosporine A favorably altered the clinical and hemodynamic course of this serious complication.


Human immunodeficiency virus (HIV) and HIV infected cells in saliva and salivary glands of a patient with systemic lupus erythematosus. A young woman with systemic lupus erythematosus (SLE) was infected with human immunodeficiency virus 1 (HIV-1) and about 6 years later developed persistent bilateral parotid gland enlargement. It was unclear whether this represented salivary gland involvement as a component of her SLE (secondary Sjögren's syndrome) or the initial clinical manifestation of her HIV-1 infection. HIV proviral DNA was found in individual salivary glandular secretions and in whole saliva. Additionally, cells positive for HIV RNA were isolated from whole saliva. A parotid gland biopsy revealed infiltrating lymphocytes containing large amounts of HIV RNA.


Vasculitis with eosinophilia and digital gangrene in a patient with acquired immunodeficiency syndrome. Fewer than 40 cases of vasculitis have been described in the setting of human immunodeficiency virus infection. We describe a patient with the acquired immunodeficiency syndrome (AIDS), a heavy smoker, who developed a syndrome of constitutional symptoms, eosinophilia and digital gangrene. Vasculitis of the digital arteries was documented by angiography. He responded to high dose corticosteroid therapy with arrest of the ischemic process. After steroids were discontinued, he suffered a relapse of the vasculitis documented by skin biopsy. In patients with AIDS with this serious, potentially steroid responsive condition, steroid therapy should be considered in spite of the preexisting immunodeficiency state.


Klebsiella pneumoniae septic arthritis of a lumbar facet joint. Only 3 cases of isolated septic arthritis of a lumbar facet joint have been reported, all due to Staphylococcus aureus. We describe a case of Klebsiella pneumoniae septic arthritis of a lumbar facet joint in an HLA-B27 positive patient.


Ostrich (Struthio camelus) immobilisation using carfentanil and xylazine and reversal with yohimbine and naltrexone. Ostriches (Struthio camelus) (n = 20) were immobilised from a helicopter by darting with a total dose of 3 mg carfentanil and 150 mg xylazine. An initial excitement phase was displayed, commencing on average at 2.67 min (S.D. 0.72) after darting, and the average time to recumbency was 4.97 min (S.D. 1.05). The average heart and respiration rates prior to reversal were 121.2 (S.D. 19.96) and 13.7 (S.D. 5.96) min-1 respectively. Reversal was achieved by the intravenous injection of yohimbine at approximately 0.125 mg kg-1 and 300 mg of naltrexone, and was uneventful. Further investigations need to be done to establish the most appropriate dosage rates for these preparations in ostriches.


Breaking strength of CO2-laser and scalpel blade incisions in the dog. Incisions were made in the skin, linea alba and intestine of dogs (n = 5) with a CO2-laser and a stainless steel scalpel blade. The procedures were carried out on Day 0, 8, 10 and 11 in order to produce 1, 2, 4 and 12-day old samples. On Day 12 the dogs were euthanized, the incisions harvested and the breaking strength of each sample recorded. No significant difference in the strength developed in the skin and intestinal wounds could be observed, whereas the incisions made with the scalpel blade in the linea alba were significantly stronger.


[Resistance of small strongyles in an equine stud in South Africa to the benzimidazole anthelmintics]. This paper is apparently the first report of resistance of helminths of equids to anthelmintics in South Africa. While a strain of Cyathostominae from an Arab horse stud near Pretoria showed greatly reduced susceptibility to benzimidazoles in faecal egg reduction tests, ivermectin was apparently still unaffected. It is suggested that integrated methods of control such as alternation of horses and ruminants on pasture and mechanical removal of faeces should be used in addition to anthelmintics.


Response to selenium supplementation of sheep grazing cultivated pastures in the Natal Midlands. The response to selenium supplementation of sheep grazing cultivated pastures was investigated on different farms in the Natal Midlands, Republic of South Africa. Over a period of one year, a significant (P < 0.01) improvement of 18.2% in live mass gain and of 13.7 and 15.8% (P < 0.05) in greasy and clean wool production, respectively, was measured in response to selenium supplementation in young ewes at the Cedara Research Station. In ewes injected with a long-acting barium selenate product, the glutathione peroxidase (GSH-Px) activity in the erythrocytes was maintained at elevated levels, above that of the control group, for almost 2 years. On 6 private farms, mature ewes were dosed approximately 6 weeks before mating, with a long-acting intraruminal selenium pellet. On 2 of the farms, a significant improvement (P < 0.05) in fertility of ewes was observed with no response to supplementation in the birth mass, lamb growth rate, or in lamb mortality. The long-acting pellets maintained elevated erythrocyte GSH-Px activity in the ewes for approximately one year. Although sheep on some farms in the Natal Midlands responded to selenium supplementation, it seems advisable to establish the selenium status of a flock before embarking on a supplementation programme. The risk of an excessive selenium intake, through the inadvertent supplying of selenium in different feed sources, has been demonstrated and should be guarded against.


An assessment of the selenium, copper and zinc status of sheep on cultivated pastures in the Natal Midlands. Blood samples were collected from sheep on 24 farms in the Natal Midlands to ascertain the selenium, copper and zinc status of sheep kept under intensive grazing conditions on cultivated pastures in this region. The mean erythrocyte glutathione peroxidase activity was 34.1 enzyme units (EU: nmol NADPH oxidised min-1 mg-1 haemoglobin). A total of 59.4% of the samples contained less than 30 EU, a level below which a marginal deficiency may exist. The mean selenium concentration in whole blood was 0.57 mumol l-1. Sixty four per cent of the samples contained less than 0.63 mumol l-1, a level below which the animals were considered to be marginally deficient. The mean copper concentration in plasma was 16.1 mumol l-1 and that of zinc 15.0 mumol l-1. On none of the farms was the copper concentration in plasma below the concentrations accepted to represent a marginal deficiency, and ewes on only one farm had relatively low plasma zinc concentrations. It is concluded that many sheep grazing cultivated pastures in the Natal Midlands may be marginally deficient in selenium. However, the selenium status in sheep should be determined on individual farms before supplementation is recommended, as external sources of selenium are often supplied, mostly unintentionally. Seasonal variations in blood selenium concentrations did not follow a specific trend, probably because the sheep were grazing cultivated pastures for most of the year and feeding practices differed from farm to farm. It is therefore suggested that the selenium status of sheep be determined at the pre-mating and pre-lambing stages rather than in a specific season of the year.


Use of a semi-quantitative sweat test in thoroughbred horses. A practical test for evaluating the sweating response to various concentrations of the specific beta 2 agonist, salbutamol sulphate, is described. The results of performing this test on horses (n = 54) considered to be "free sweaters", horses (n = 6) that showed signs of heat stress following exercise, and horses with complete anhidrosis (n = 2) are presented. The results indicate that intradermal injections of 0.1 ml of salbutamol sulphate at dilutions of 10(-7) or less are suitable stimuli to elicit a visually detectable local sweating in horses with a normal sweating response. Horses that only sweated at the sites where salbutamol solutions with dilutions of between 10(-4) and 10(-6) were injected, probably had a reduced sweating response and could be considered to be suffering from partial anhidrosis. Horses with severe anhidrosis of long duration did not sweat, even in response to salbutamol solutions with dilutions of 2 x 10(-3).


A quantitative study of the attitudes of members of a black developing community towards their dogs and veterinary services. To further the study of human-companion animal interactions in black developing communities, the clients (n = 94) of a university-based animal hospital, serving primarily a developing community, were interviewed. The interviews covered demographics, socio-economic aspects, perceptions, values and attitudes of clients towards their dogs. Regular and new clients were compared. A profile of the hospital's black clientele was also constructed. The study showed that the majority of black clients value their dogs for the sake of security and protection. Attitudes towards pets parallelled improvements in socio-economic status. Most clients felt positive towards existing veterinary services. The results of this study support and highlight the value of a previous observational study and could prove to be invaluable in designing, implementing and planning socially relevant animal health services.


Anion-dependent transport of thallous ions through human erythrocyte membrane. Undirectional fluxes of 204Tl+ through the human red blood cell membrane were measured. The inward rate coefficient measured in a K(+)-free saline was 15.6 +/- 0.6 hr-1. The influx of Tl+ could be partially inhibited with 0.1 mM ouabain (by 28%), 0.1 mM DIDS (by 50%) or 1 mM furosemide (by 51%). The inhibitory effects of ouabain and DIDS or furosemide were additive. Half-maximal responses were seen at 0.72 microM and 0.22 mM concentrations of DIDS and furosemide, respectively. A similar action of these blockers on Tl+ influx was observed in the erythrocytes incubated in MgCl2-sucrose media. The outward rate coefficient of 204Tl was also inhibited by DIDS and furosemide (by 65 and 52%, respectively). Rate coefficients of 204Tl influx and efflux decreased significantly in the red cells exposed to Cl(-)-free media (NaNO3 or Mg(NO3)2-sucrose). Under these conditions addition of DIDS and furosemide led to only a small inhibition of Tl+ fluxes. There was a linear increase in Tl+ influx with rising of external Cl- concentration within 80-155 mM or HCO3- concentration from 20 to 40 mM when the sum of anions was kept constant (155 mM) with NO3-. The HCO3(-)-stimulated Tl+ influx was completely blocked by 0.05 mM DIDS but only 67% by 1 mM furosemide. The present study provides direct evidence for the occurrence of Cl- (HCO3-)-dependent, DIDS-sensitive movement of Tl+ across the human erythrocyte membrane in both directions. Under physiological conditions, about half of net Tl+ fluxes occurs due to an anion exchange mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)


The permeability of reconstituted liposomes containing the purified lens fiber cell integral membrane proteins MP20, MP26 and MP70. A number of lens fiber cell integral membrane proteins have been localized to junctional regions where they have been proposed to play a role in either mediating or controlling cell-to-cell communication. We have examined the effect of three lens fiber cell membrane proteins, MP20, MP26 and MP70, on the permeability properties of unilamellar phospholipid liposomes. This approach has been previously used to examine the channel-forming properties of MP26. Liposome permeability was determined by measuring the effect of Co2+ on the quenching of the fluorescence of N-4-nitrobenzo-2-oxa-1,3 diazole phosphatidyl ethanolamine (NBD-PE)-containing liposomes as described previously by Scaglione and Rintoul (Invest. Ophthalmol. Vis. Sci. 30:961-966, 1989). The effect of all three proteins on liposome permeability was similar. Permeability was dependent on the protein/phospholipid ratio and was not significantly affected by agents known to modify gap junctional permeability in vivo. Glycophorin A, a non-channel-forming integral membrane protein derived from erythrocytes, was also shown to increase the permeability of unilamellar phospholipid liposomes. The ability of a non-channel membrane protein to increase Co2+ quenching of NBD-PE-containing liposomes (presumably in a nonspecific manner) indicates that reports describing the permeability of lens membrane protein-containing liposomes should be interpreted with caution in terms of their relationship to cell-to-cell communication.


Transvaginal sonography of cervical width and length during pregnancy. Transvaginal ultrasonography has been proposed as a reliable method of assessing dimensions of the cervix. The purpose of the current investigation was to establish normative data for cervical width and length during pregnancy. This information may be helpful in predicting patients at risk for preterm birth. A transvaginal 5 or 7.5 MHz transducer was used on 132 consecutive low-risk pregnant women undergoing evaluation for gestational dating purposes. Even in cases of patient obesity or an empty bladder, high resolution was possible and permitted cervical measurement in all but one case. The external cervical width at the vaginal fornices was found to increase with advancing gestation (R = 0.512, P < 0.005). The cervical length from internal os to external os was found not to change with advancing gestation (R = 0.11, P = 0.30). Using these normative data, investigation is recommended to determine whether measuring cervical width and length is useful in predicting preterm labor and delivery.


G-protein activators induce a potassium conductance in murine macrophages. The whole-cell patch clamp technique was used to test whether intracellular application of G-protein activators affect ionic currents in murine macrophages. Both the J774.1 macrophage-like cell line and primary bone marrow derived macrophages were used. Cells were bathed in Na Hanks' solution and intracellularly dialyzed (via the patch pipette) with K Hanks (145 mM KCl, < 100 nM Ca) plus or minus the G-protein activators GTP gamma S (10 microM), GppNHp (10 microM), or AIF4- (200 microM AlCl3 + 5 mM KF). In the absence of G-protein activators, only two K currents, an inwardly rectifying K current (Kir) and an outward, inactivating K current (Ko) were observed. In the presence of protein activators, two effects were observed: (i) the Kir conductance, which is stable for up to 30 min under control conditions, decayed twice as fast and (ii) an outwardly rectifying, noninactivating current appeared. The induced outward current appeared < 2 min after attaining the whole-cell patch clamp configuration. The current could be distinguished from the Kir and Ko currents on the basis of its direction of rectification (outward), barium sensitivity (> 1 mM), and kinetics (no time-dependent inactivation). Intracellular application of GTP (500 microM), GDP (500 microM), cAMP (100 microM + 0.5 mM ATP), or IP3 (20 microM) did not induce the current; 100 microM ATP gamma S activated a half-maximal amount of current. Induction of outward current by 10 microM GTP gamma S could be prevented by pre-exposing cells to pertussis toxin but not cholera toxin. This current is K selective since (i) its induction was accompanied by hyperpolarization of the cell toward EK, even after Kir had "washed out", (ii) it was present after > 90% of both intracellular and extracellular Cl were replaced by isethionate, and (iii) the induced outward conductance was absent when Ki was completely replaced by Cs, and was reduced by approximately 1/3 when [K]i was reduced by 1/3. Quinidine (1 mM) and 4-aminopyridine (10 mM) inhibited the current, but apamin (1 microM) and charybdotoxin (1 microM) did not.


Inability of color and spectral Doppler to identify fetal renal obstruction. Color flow Doppler imaging was used to obtain fetal renal artery flow velocity waveforms in 130 normal fetuses at various gestational ages to build a database for the establishment of normal ranges for the Pulsatility Index in the fetal renal arteries throughout pregnancy. Twenty-nine cases of fetal renal tract dilation (greater than 5 mm anteroposterior diameter of the renal pelvis) of various causes were investigated. We found that measurements of the pulsatility index were not significantly altered from the normal range, so that color flow Doppler does not appear to be helpful in the differential diagnosis of fetal renal dilation.


Abnormal spinal curvature in the fetus.

Abnormal spinal curvature in the fetus. We reviewed 20 cases of sonographically detected abnormal fetal spinal curvature to help determine the significance of this finding. Both marked (13 cases) and mild (7 cases) curves were diagnosed sonographically. Sonography showed associated anomalies in 19 fetuses, including neural tube and ventral wall defects. Outcomes were poor, with only three infants surviving. Isolated scoliosis was identified prenatally in one neurologically normal infant with hemivertebrae. We conclude that abnormal spinal curvature in the fetus is a significant finding, whether mild or severe. Even as an isolated finding, it is significant, as prenatal detection assists postnatal management.


Evaluation of celiac and mesenteric vascular disease with duplex ultrasonography. A retrospective study of 25 patients was performed to evaluate the applicability of duplex ultrasonography to the celiac and superior mesenteric circulation. Lateral contrast aortograms were compared to the pulsed-Doppler spectral data from duplex examinations in fasting subjects. A significant correlation was identified between the celiac-aortic diastolic ratio and the degree of stenosis measured angiographically, but such a relationship could not be established for the superior mesenteric artery. These data suggest that duplex ultrasonography may not be a reliable screening test for mesenteric vascular disease.


Compression ultrasonography. Treatment for acute femoral artery pseudoaneurysms in selected cases. Treatment of pseudoaneurysms has traditionally been surgical. We evaluated 12 patients who developed femoral artery pseudoaneurysms after cardiac catheterization. Using color Doppler ultrasonography steady pressure was applied, occluding flow in both the neck and body of the pseudoaneurysm for 10 minute intervals. Six patients were treated successfully. We believe the same technique was unsuccessful in five additional patients because compression of a short (< 5 mm) or wide neck is ineffective. In one patient the pseudoaneurysm thrombosed spontaneously prior to a treatment attempt. We believe compression ultrasonography of pseudoaneurysms should be offered as a therapeutic alternative to surgery in selected cases.


[Molecular biological approaches in nephrology]. Recent explosive advances in molecular biological techniques have greatly expanded their applications, and they are now widely employed in nephrology. Main applications of molecular biological techniques in the field of nephrology are 1) molecular cloning of genes coding important proteins, 2) analysis of gene expressions, 3) approach using transgenic animals, 4) molecular genetic approach for inheritable diseases such as adult polycystic kidney disease. These techniques are now considered to be one of the most important and powerful tools in the field of nephrology.


[Application of transgenic model in renal research]. In the present report, we have used transgenic models to analyze the role of growth factors in the development of glomerulosclerosis. All peptides inducing a growth response do not lead to glomerulosclerosis, however, several transgenic models including growth hormone, transforming growth factor-a or SV 40T antigen developed progressive glomerulosclerosis, which was associated with glomerular hypertrophy. Evidence obtained from these transgenic models suggests that glomerulosclerosis may be related to a dysregulation of glomerular cell growth and a synthesis of extracellular matrix components by resident glomerular cells.


[In situ hybridization histochemistry for the kidney diseases]. Crucial points of in situ hybridization methods were pointed out for study on the kidney diseases at molecular level. As examples of in situ hybridization study on kidney, we introduced two cases: 1) expression of the hepatocyte growth factor gene during renal regeneration after unilateral nephrectomy, and 2) induction of the metallothionein gene expression by ischemic acute renal failure. We proposed an idea to establish a probe bank for the in situ hybridization study.


[Usefulness of microdissection of nephron segments and fluorescent indicator for molecular biological studies of nephron functions]. The kidney consists of numerous functional units called nephrons. Thus, the use of individual nephron segments is essential to characterize their functional properties and to clarify the molecular basis of site-specific functions. Nephron segments can be microdissected from collagenase-treated renal slices under a stereomicroscope. A variety of intracellular ionic concentrations or membrane potential can be determined with various fluorescent probes. Fura-2/AM-loaded nephron segments reveal a transient increase of cytosolic free calcium concentrations by agonists such as angiotensin II, vasopressin, kinins, etc. To localize their receptors or to characterize their subtypes, this technique is especially beneficial, because tiny fragments of the nephron are sufficient by combination with a two-wave length microscope fluorometer. As an example, discovery of a novel vasopressin receptor (Vp) is described.


[Renin-angiotensin system].

[Renin-angiotensin system]. Renin is a key enzyme in the renin-angiotensin system (RAS) which controls the blood pressure and electrolyte balance. By numerous studies on RAS and structures of renin and prorenin, the determination of three-dimensional structure for renin has been thought to be important in the rational approach to the drug design for antihypertensives. Here, several investigations for the three-dimensional structure of human and mouse renins are reviewed. These papers showed that the active site cleft had a less open arrangement in renins than that in other aspartic proteinases, although the general topology of both renins were quite similar. They also described subsites of renins and others.


[Molecular biology of the natriuretic peptide system]. The natriuretic peptide system is composed of at least three distinct endogenous peptides: atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and C-type natriuretic peptide (CNP), and three receptors: ANP-A receptor/GC-A, ANP-B receptor/GC-B and the clearance receptor. This system influences the control of body fluid and blood pressure as cardiac hormones and local regulators. Recent advances in molecular biology have unravel the molecular mechanism of the natriuretic peptide system and have facilitated our understanding of it. The present review gives the current knowledge of the molecular biology of the natriuretic peptide system.


[Interleukin 6 (IL-6)].

[Interleukin 6 (IL-6)]. Interleukin 6 (IL-6) is a polypeptide mediator regulating immune response, acute phase reaction and hematopoiesis. Disregulating production of IL-6 was suggested to be involved in a variety of chronic inflammatory diseases. IL-6 is an autocrine growth factor for cultured mesangial cells. IL-6 was localized in the mesangial area of the kidney tissue and intraglomerular expression of mRNA, encoding for IL-6 gene, was observed in patients with mesangial proliferative glomerulonephritis (mes PGN). Furthermore, mesangial proliferation was observed in IL-6 transgenic mice. These results suggest that IL-6 is closely associated with the pathogenesis of mesPGN.


[Growth factors: a regulator of renal function]. We selectively focus on two growth factors, epidermal growth factor (EGF) and insulin-like growth factor (IGF), and discuss their roles on regulation of renal function and associated diseases conditions, as well. EGF, 6 kD polypeptide, is derived by proteolysis from a large precursor (prepro EGF, 133 kD). Prepro EGF is a membrane-anchored protein and its mRNA is predominantly localized to distal tubules of mouse and rat kidneys. We immunohistochemically demonstrated the glomerular distribution of EGF and EGF-receptor in normal and nephritic human kidneys. The physiologic roles of EGF produced in the kidney are various; it is mitogenic for tubular epithelial cell, inhibits gluconeogenesis and salt and water reabsorption in the tubules, and effects on glomerular hemodynamics. Alteration of renal EGF expression is suggested in renal ischemic injury, renal hypertrophy and cystic renal disease. IGF-I, somatomedin-C, is produced in collecting duct, and glomerular cells, and exerts a variety of actions on kidney. IGF-I stimulates gluconeogenesis in renal tubules, and is mitogenic for mesangial cells. The administration of this growth factor increases glomerular filtration rate. Enhanced expression of renal IGF-I was observed in the uninephrectomized animals.


[The molecular basis for the steroid hormone actions in the kidney]. Steroid hormones such as mineralo- and glucocorticoids act as important regulators for the kidney to maintain the body fluid homeostasis. During the past 40 years, numerous investigations have been dedicated to the elucidation of precise mechanisms of steroid hormone actions in their target tissues including the kidney. Now it is well recognized that cellular actions of steroid hormones can be accomplished by two step processes, namely a specific receptor binding in the cytosol and/or the nucleus and a new protein synthesis through a gene transcription. The efforts for the isolation and purification of steroid hormone receptors have been also made and followed by the great successes of molecular techniques in the cDNA cloning and determination of amino acid sequences for them in recent years. The revealed high homology of the functional domains of receptors has renewed the issues of how tissue specificities of steroid hormone actions are determined. The identification of steroid-induced proteins and cellular steroid hormone metabolites would be also important issues to be studied.


[Adenosine and adenosine receptors in the kidney]. In the kidney, adenosine plays important regulatory roles, including renal blood flow, glomerular filtration rate, renin secretion, tubuloglomerular feedback, tubular reabsorption of sodium and water, sympathetic neurotransmitter release, and erythropoietin secretion. These functions are mediated through adenosine 1 (A1)-receptors and adenosine 2 (A2)-receptors. These receptors couple to the inhibition and stimulation of adenylate cyclase, through Gi and Gs proteins, respectively. A variety of other effecter systems have been reported to be coupled to A1 receptors, including phospholipase C, phospholipase A2 and potassium, as well as Ca++ channels. Recently, A1 receptors, A2 receptors and novel A2 receptor have been cloned, sequenced and expressed. In association with the development of selective adenosine analogues, we are now ready to take up problems at the biochemical and molecular biological levels.


[Molecular structure and function of renal adrenergic receptors]. The kidney is the principal organ in the body that mediates the excretion of water and minerals, and also functions as an endocrine organ in producing hormones, enzymes (e.g., renin), and vasoactive molecules (e.g., prostaglandins). Circulating catecholamines and autonomic innervation of the kidney can influence those important functional responses as well as vascular constriction via renal adrenergic receptors. The adrenergic receptors are members of a large family of G protein-coupled receptors. Recent advance of molecular cloning has revealed a growing heterogeneity of adrenergic receptor subtypes. They share a common topographical motif consisting of seven putative transmembrane regions, where. They are thought to be essential for ligand binding. Multiple subtypes of adrenergic receptors may exist in the kidney. Defining the precise role of renal adrenergic receptors in disease states, particularly hypertension, will certainly require the application of new tools of biochemistry and molecular biology.


[Molecular pharmacology in diuretics].

[Molecular pharmacology in diuretics]. Recently, much knowledge of the molecular mechanism of diuretic actions has been accumulated. Molecular cloning of carbonic anhydrase revealed a difference in isozymes and the localization of each isozymes was also identified. Carbonic anhydrase VI, located in brush border of the proximal tubules, is inhibited by acetazolamide. Loop diuretics, furosemide, bumetanide, piretanide and ethacrynic acid, inhibit Na(+)-K(+)-Cl(-)-cotransport system in the thick ascending limb of the Henle's loop. The histochemical localization and biological characterization were investigated by using [3H]-bumetanide. Loop diuretics cotransport protein and inhibit Na+ and Cl- reabsorption. Spironolactone inhibits the binding of aldosterone and cytosol mineralocorticoid receptor in the cortical connecting tubule competitively.


[Regulation of fibrillar and type IV collagen genes]. Control of collagen synthesis can be exerted through control of mRNA levels by regulating of gene transcription and RNA maturation or the stability of mRNA. For fibrillar collagen genes many of the cis-acting regulatory elements are located in a relatively small region 5' to the transcription start site. The exon structure of the type IV genes differs significantly from that of fibrillar collagen in that triple-helical exons show considerable divergence from the 54 bp motif. The first intron of the alpha 1 (IV) is shown to contain an enhancer that can activate promoter. To identify the cellular mechanisms that determine the tissue-specific expression of the different collagen genes, the mechanisms whereby these genes respond to various cytokines and hormones that influence their expression.


[Abnormality of type IV collagen metabolism in the development of diabetic nephropathy]. Diabetic nephropathy is characterized by the thickening of glomerular basement membrane and the expansion of mesangium. These changes are considered to be caused by an accumulation of the extracellular matrix protein including type IV collagen. The mesangial cells are the cells responsible for the synthesis and metabolism of type IV in the mesangial cells are discussed. The role of advanced glycation end products in the pathogenesis of diabetic nephropathy will be also discussed.