Multiple regression analysis of twin data: a model-fitting approach.

The multiple regression methodology proposed by DeFries and Fulker (DF; 1985, 1988) for the analysis of twin data is compared with maximum-likelihood estimation of genetic and environmental parameters from covariance structure. Expectations for the regression coefficients from submodels omitting the h2 and c2 terms are derived. Model comparisons similar to those conducted using maximum-likelihood estimation procedures are illustrated using multiple regression. Submodels of the augmented DF model are shown to yield parameter estimates highly similar to those obtained from the traditional latent variable model. While maximum-likelihood estimation of covariance structure may be the optimal statistical method of estimating genetic and environmental parameters, the model-fitting approach we propose is a useful extension to the highly flexible and conceptually simple DF methodology.


Genetic and environmental effects on type A scores in monozygotic twin families.

Monozygotic (MZ) twin pairs with spouses and children, altogether 787 subjects, completed the Jenkins Activity Survey (JAS). The observed correlations for the various sets of relationships fitted well with biometric models including only parameters for additive genetic effects and, for Type A and Job Involvement, assortative mating. There was no evidence of effects of the family environment (cultural transmission) or genetic dominance (nonadditivity). For all but the Hard Driving and Competitive scale, there was evidence of effects of sex-specific genes. The heritability estimates were, for males and females, respectively, .33 and .39 for Type A, .36 and .48 for Job Involvement, .20 and .52 for Speed and Impatience, and .13 (both sexes) for Hard Driving and Competitive. The estimates given here are deflated by measurement errors and should probably be corrected by multiplying by values in the neighborhood of 1.3. Even after correction, the results suggest that individual differences for Type A and related traits depend more on nonfamilial environment than on genes.


Mouse models of human single gene disorders. I: Nontransgenic mice.

Mouse models of human genetic disorders provide a valuable resource for investigating the pathogenesis of genetic disease and for testing potential therapies. The high degree of resolution of linkage mapping in the mouse allows mutant phenotypes to be mapped precisely which, combined with the accurate definition of areas of homology between the mouse and human genomes, greatly facilitates the identification of mouse models. We describe here mouse models of human single gene disorders dividing them into three categories depending on the information available; phenotypic similarities, comparative mapping and identification of the underlying genetic lesion.


The molecular biology of desmosomes and hemidesmosomes: "what's in a name"?

Desmosomes are junctions involved in intercellular adhesion of epithelial cells and hemidesmosomes are junctions involved in adhesion of epithelia to basement membranes. Both are characterised at the ultrastructural level by dense cytoplasmic plaques which are linked to the intermediate filament cytoskeleton of the cells. The plaques strongly resemble each other suggesting a relationship between the two kinds of junctions, as implied by their names. Recent characterisation of the molecular components of the junctions shows they are, in fact, quite unrelated implying that structural similarity is fortuitous. The molecular biology raises many fascinating problems relating to their structure and function.


The molecular basis of the type 1 glycogen storage diseases.

Microsomal glucose-6-phosphatase catalyses the last step in liver glucose production. Glucose-6-phosphatase deficiency, now termed type 1 glycogen storage disease, was first described almost 40 years ago but until recently very little was known about the molecular basis of the various type 1 glycogen storage diseases. Recently we have shown that at least six different proteins are needed for normal glucose-6-phosphatase activity in liver. Four of the proteins have been purified and three cloned. Study of the type 1 glycogen storage diseases has stimulated investigations of the mechanisms of small molecule transport across the endoplasmic reticulum membrane and demonstrated the existence of novel endoplasmic reticulum transport proteins for glucose and phosphate.


A mammalian molecular clock?

The molecular clock hypothesis remains controversial more than a quarter of a century after it was first proposed. A variety of approaches have been applied to testing the molecular clock in mammals. In many of these studies apparent refutation of the molecular clock has been based on false assumptions about the pattern of mammalian evolution. With a few exceptions there now appears to be little evidence for variation in the rate of molecular evolution among mammalian lineages, although comparison of more genes and proteins among a greater variety of mammalian orders is needed before a definitive conclusion can be reached on the issue.


Carrier detection of deletions in female relatives of X-linked disorders by non-isotopic in situ hybridisation.

Recent studies suggest that a non-isotopic in situ hybridisation (NISH) approach can be successfully employed to investigate the carrier status of female relatives in families of selected patients with Duchenne muscular dystrophy (DMD) or Hunter syndrome, whose diseases are due to a specific X chromosome deletion. Whilst the majority of metaphase spreads from normal females show specific hybridisation signals on both X chromosomes when tested with either dystrophin or Hunter gene-derived probes, only one X chromosome in each metaphase spread will show the relevant hybridisation complex in female carriers of deletions involving the dystrophin or Hunter gene. Thus, the NISH method can be a valuable diagnostic tool for the detection of the carrier status of female relatives of patients with X chromosome deletions.


Two proteins with ribosome-inactivating, cytotoxic and abortifacient activities from seeds of Luffa cylindrica roem (Cucurbitaceae).

Two immunologically distinct glycoproteins, with a molecular weight of 28,000 and 28,500 respectively, were isolated from seeds of Luffa cylindrica using a procedure that involved acetone precipitation, gel filtration on Sephadex G-75 and ion exchange chromatography on CM-Sepharose CL-6B. The proteins differed in the content of aspartic acid, threonine, proline and alanine but were otherwise similar in amino acid composition. The proteins were capable of inducing mid-term abortion in mice, inhibiting protein synthesis in a cell-free system and suppressing thymidine uptake by human choriocarcinoma cells.


Novel fractionation of proteins using novobiocin as a precipitating agent: application to ribosomal proteins from the archaebacterium Sulfolobus acidocaldarius and Escherichia coli.

A fractionation method involving precipitation of proteins with novobiocin has been developed to separate ribosomal proteins of the archaebacterium Sulfolobus acidocaldarius. Four groups of proteins were obtained corresponding to those precipitated with 0.2, 0.5 and 1.0 mg/ml novobiocin and the proteins soluble at 1.0 mg/ml novobiocin. The same procedure was also successful for the fractionation of ribosomal proteins from Escherichia coli. Two dimensional gel electrophoresis showed that proteins present in each group were distinct with only a minor contamination of proteins from other groups. Our results indicate that proteins with similar charge and molecular weight can be separated by this method. This procedure may have applications for the fractionation of other complex protein mixtures.


Purified uteroglobin lacks anti-proteinase activity.

Rabbit uteroglobin was purified from both uterine fluids and lung lavages by a combination of gel filtration and ion-exchange column chromatography. Anti-trypsin and anti-papain activities were measured in the fractions of the eluates. Anti-proteinase activities were detected in minor contaminants eluting close to the uteroglobin peak but the protein itself was devoid of anti-proteinase activity. Ion-exchange-purified uteroglobin also lacked inhibitory activity of elastase, chymotrypsin or subtilisin. The presence of contaminants could explain the anti-proteinase activity reported occasionally for uteroglobin.


Reconstitution of a purified and partially active low Km iodothyronine 5'-deiodinase with phospholipids.

We previously reported the detergent-extraction and purification to homogeneity from rat liver microsomes of an iodothyronine 5'-deiodinase which differed from the type I enzyme in its molecular size as well as its Km, thiol responsiveness, and its expected sensitivity to the antithyroid drug, propylthiouracil. The specific activity enhancement (22-fold) of the delipidated enzyme was, however, relatively modest. We report here that the purified enzyme can be activated a further 25-fold by reconstitution in microsomal phospholipids and, to a lesser extent, in soybean phospholipids. The data suggest that a native lipid environment is necessary for proper conformation of the enzyme for optimal activity.


Bovine erythrocyte glutathione S-transferase: purification, inhibition, and complex formation.

Glutathione S-transferase has been purified from bovine erythrocytes by affinity chromatography. The enzyme has an isoelectric point of 7.2, behaves as a 48-kDa protein composed of two identical subunits, and has an N-terminal sequence of PPYTIVYFPVQGR?EAMRMLL. This sequence, the amino acid composition, and the kinetic parameters suggest that the enzyme belongs to the pi-class of transferases. Hemins, porphyrins, and fatty acids form complexes with the enzyme and serve as effective inhibitors. Treatment of the transferase with N-ethylmaleimide, 3-amino-1,2,4-triazole, diethyl pyrocarbonate, or 2,3-butanedione inhibits transferase activity without altering tetrapyrrole binding. The role of the complexation and inhibition of glutathione S-transferase in erythroid metabolism has yet to be elucidated.


Purification and characterization of the HPR protein of Pediococcus halophilus.

HPr protein, the ptsH gene product, of the phosphoenolpyruvate:sugar phosphotransferase system of Pediococcus halophilus was purified to homogeneity using heat and acid treatments, DEAE-Sepharose CL-6B and hydroxyapatite column chromatographies. The purified protein complemented the HPr activity of the phosphotransferase system in Staphylococcus aureus ptsH mutant cell lysate. The molecular weight was estimated as 6,500 by SDS polyacrylamide gel electrophoresis. The amino acid sequence of the amino-terminal part (1-44) of the native purified protein was highly homologous to those of HPr proteins from gram-positive bacteria. Antiserum raised against the purified HPr protein specifically reacted with the Pediococcus halophilus HPr protein and did not cross-react with Staphylococcus aureus or Escherichia coli HPr proteins.


A tentacle gel simplifies the purification of ceruloplasmin.

The mechanism of how chloroethylamine-treated agarose greatly simplifies the purification of ceruloplasmin (Cp) by preferentially binding this protein from blood plasma has been investigated. Chloroethylamine readily cyclizes to ethylenimine under alkaline conditions which then polymerizes to polyethylenimine (PEI). Ethylenimine and PEI were detected in the reaction mixture used to generate the resin. PEI polymers are grown from the matrix and form "tentacle" ligands, while PEI-silica and PEI-cellulose which are not tentacle gels did not bind Cp as readily. Spermine and spermidine, naturally occurring polyamine compounds, were attached to CNBr activated-agarose and showed weak affinity for Cp. [It should be noted that ethylenimine is a potential human carcinogen and presents an inhalation hazard.]


Rate laws for the exchange of guanine nucleotides bound to EF-Tu catalysed by EF-Ts.

The rate laws governing the exchange of guanine nucleotides bound to EF-Tu catalysed by EF-Ts are set out and the information that is available from various studies is analysed in terms of the laws. In particular the consequences of k-1 being equal to k-4 in the reaction sequence as set out below are examined in relation to results obtained by Hwang and Miller (J. Biol. Chem. 260, 11498-11502) and the contradiction these pose in relation to other published results. Measurement of on and off rate constants for binding of GTP to EF-Tu.EF-Ts and of the affinity of EF-Ts for EF-Tu.GTP should prove of importance in resolving discrepancies.


Isolation and purification of mitochondrial monoamine oxidase B from bovine liver by chromatofocusing.

Mitochondrial monoamine oxidase B has been isolated and purified from bovine liver. The isolation procedure involves (a) a series of purification steps including isolation of the mitochondria, extraction with non-ionic detergents and chromatofocusing; (b) a novel fluorimetric titration assay. The purified enzyme is isolated following a 8.8 fold purification with an overall yield of 12% and a specific activity of 3460 nM.g-1. A comparison is made with other established procedures. Ion exchange chromatography on DEAE-Cellulose produced a 6.8 fold purification, 6.2% yield and 2383.3 nM.g-1 specific activity while with affinity chromatography on Blue Sepharose CL-6B gave a 3 fold purification, 4.1% yield and a specific activity of 1152.7 nM.g-1.


Membrane binding and autolytic activation of calpain-I in human platelets.

The binding of calpain-I (Ca2+ activated neutral protease with high Ca2+ sensitivity) to the membranes of human platelets and the subsequent autolytic activation of calpain-I were analyzed by an immunoblot technique. In A23187 stimulated platelets, cytosolic calpain-I translocated to the membranes with autolysis in a Ca2+ dependent manner and simultaneously underwent autolysis. Although calpeptin, a cell permeable calpain inhibitor, inhibited autolysis of calpain-I, it was unable to prevent the translocation of calpain-I. In a cell re-constituted system, the membrane binding of calpain-I was also Ca2+ dependent and was significantly inhibited by a substrate of calpain. It was suggested that the binding of calpain-I to the membranes required the substrate binding site of this enzyme.


[Protein motifs and their structure-functional role].

The review deals with repeating fragments of amino acid sequences, so-called "motifs", that are important in maintaining structural integrity and/or function of various proteins, especially those interacting with phospholipid aggregates. The occurrence of Phe-Leu-Gly motif characteristic for the amino acid sequence of the primate immuno-deficiency viruses fusion peptides is analysed in various proteins, as well as tripeptide fragments of general formula Xaa-Xah-Gly (Xaa-Phe, Tyr; Xab-hydrophobic amino acids Ala, Val, Leu, Ile) homologous to the above motif and retro-sequences Gly-Xab-Xaa. These tripeptide repeats are characteristic for the amino acid sequences of complex membrane proteins, viral envelope proteins, proteinases and proteins connected with energy transfer or interacting with lipids. These repeats are frequently met in conservative regions of amino acid sequences, in sites readily accessible for other molecules at the boundary of or between structured fragments, this being due to the backbone semi-coiled form's preference in the given amino acid fragment. This protein motif appears to play an important role at the initial stages of the large protein's interaction with the phospholipid membrane.


[Conformational aspects of the biological action of functional fragments of the p21ras oncoprotein family. 1. Fragments 1-11 and 9-16 of the polypeptide chain].

Using theoretical conformational analysis, spatial structures of the N-terminal undecapeptide, common to all p21 modifications, and of the 9-16 fragments of the protein's active and passive analogues have been investigated. The data obtained reveal an essential differences between the predominant backbone forms of the active and passive modifications of the oncoprotein.


[Conformational aspects of the biological effect of functional fragments of the p21ras oncoprotein family. 2. Fragment 1-16, various sequences].

The conformational analysis data on active ([Val12-Gly13], [Asp12-Gly13] and [Gly12-Asp13]) and passive ([Gly12-Gly13] and [Pro12-Gly13]) modifications of the p21ras family oncoproteins are presented. The activating amino acid substitutions are shown to be accompanied by essential changes in the secondary structure, resulted in the 9-16 fragment spiralization. The spatial structure of the 1-9 fragment does not vary for all the predominant forms of the active and passive analogues. The results of the conformational analysis have been used for studying the structural-functional relationships.


[Effective complementarily-addressed photomodification of nucleic acids by oligonucleotide derivatives, containing aromatic azido groups].

Highly effective site-specific photomodification of a DNA-target was carried out with oligonucleotide reagents carrying aromatic azido groups. Oligonucleotide derivatives with a photoactive function R on the 5'-terminal phosphate and at C-5 atom of deoxyuridine were synthesized: R1NH(CH2)3NHpd(TCCACTT) and d(ULNHRCCACTT), where R1 is p-azidotetrafluorobenzoyl, R2 is 2-nitro, 5-azidobenzoyl, R3 is p-azidobenzoyl; LNH = -CH2NH-, -CH2OCH2CH2NH- or -CH2NHCOCH2CH2NH-. The prepared compounds form stable complementary complexes and effect site-specific photomodification of the target DNA. The modification of pentadecanucleotide d(TAAGTGGAGTTTGGC) with the reagents was investigated. Maximum extent of modification strongly depended on the reagent's type, the photoreagent with R1 being the most effective. Whatever the binding site was, this agent provided a 65-70% modification in all cases except LNH = -CH2NH-, when the yield was twice lower. For the reagents bearing R1 the modification sites were identified. Selective modification at the G9 residue was detected in the case of LNH = -CH2OCH2CH2NH- and when a photoactive group was linked to the terminal phosphate.


[Dependence of anti-acetylcholinesterase effectiveness of phosphoorganic inhibitors on the accessibility of the phosphorus atom].

All equilibrium conformations of 12 anti-acetylcholinesterase organophosphorus inhibitors were calculated by the molecular mechanics method. The accessibility of the phosphorus atom of the inhibitors for interactions with the nucleophilic group at the enzyme active centre was estimated. The conformers with the phosphorus atom sterically accessible from the side opposite to the breaking ester bond were classified as productive. A correlation was revealed between the activity of the inhibitors and the population of their productive conformation.


Low birth weight in the central Canadian Arctic.

Birthing, health and prenatal records of 523 infants born between 1984 and 1988 in the central Canadian Arctic, were reviewed. The incidence of low birth weight (LBW) was 5.5%, which is lower than in many other populations. Principal indicators of risk were a previous history of a LBW infant, a short inter pregnancy interval, low weight at the time of the first prenatal visit and a weight gain of less than 200 gm/wk throughout pregnancy. Use of nicotine and alcohol may have been contributing factors. Although low, it is argued that the incidence could be further reduced through prepregnancy social and educational programs, as well as intensified prenatal care for those at risk.


Infant mortality in Greenland: secular trend and regional variation.

Infant mortality and stillbirths were studied in the Inuit population of Greenland. Infant mortality rate decreased from 110 per 1000 live births in 1951 to 50 in 1970 and 20 in 1989. Statistically significant decrease was in particular seen for acute respiratory infections and certain perinatal conditions. Both perinatal and postperinatal mortality rates increased significantly with remoteness from the capital and were higher in settlements than in towns. The overall trend in Greenland is similar to the Danish pattern 30 years earlier, but there was pronounced regional variation. In towns, the infant mortality rate decreased continuously while in settlements an initial decrease was followed by a substantial increase during 1980-87 when the mortality rate doubled. The greater part of this increase could be attributed to an increased number of deaths in the postperinatal period countrywide during 1981-84 and an increased number of perinatal deaths in the settlements of one particular municipality during 1983-86.


Influence of genetic and demographic factors on etiology and pathogenesis of chronic disease in north Siberian aborigines.

The relationship between genetico-demographic parameters (endogamy, inbreeding, mating structure, gene diversity) and regularities of the prevalence of hereditary and common diseases (ischemic heart disease, arterial hypertension) and their risk factors in the North Khanty, a small, semi-isolated Siberian ethnic group of Finno-Ugric linguistic affinities, were studied. It has been shown that among hereditary diseases autosomal-dominant and polygenic ones are most frequent. Special features of the genetic liability to common diseases were described. The contributions of hereditary factors to the total variability of the 20 studied morphophysiological traits were shown to vary significantly.


The role of hereditary factors in phenotypic variability of hormone levels in the population genetically adapted to circumpolar environment.

The variability of the plasma cortisol, T3, T4, TSH and estradiol levels were studied in the North Khanty population genetically adapted to the extreme conditions of the North. The relationships between genetic markers and hormone concentrations were also studied. Negative linear correlations of T3 and T4 levels with age were obtained. Cortisol, estradiol and T3 levels showed nonlinear age dependency. T4 concentrations were significantly lower in August than in February and November. The highest heritability estimate was shown for estradiol levels (44%).


The death divide: differentials in premature mortality by gender in Puerto Rico.

The use of the indicator "years of potential life lost" (YPLL) highlights the extent to which premature mortality in Puerto Rico is a predominantly male phenomenon. While men accounted for 58.6% of all deaths in 1987, they represented fully 71.8% of all YPLL attributed to the thirteen leading causes of death. The breakdown of YPLL by gender also underlines sex-specific differences in the causes of mortality. While accidents constitute the leading cause of premature death among men, malignant neoplasms take the lead among women. Similarly, homicides and cirrhosis are significant sources of years of life lost among males, while pneumonia/influenza and diabetes are higher priorities among females. These findings suggest that health promotion strategies need to be gender-specific in order to reach the right targets.


The new macrolides: expanding the ways in antibiotic treatment.

After almost forty years of its introduction, erythromycin will not be the exclusive member of the macrolide group of antibiotic agents, but a new generation of its derivatives which surpass it in pharmacological properties and clinical efficacy will also be available. Clarithromycin, a 14-membered derivative, has shown acid stability, longer half-life, lower protein binding and higher lung tissue penetration. Its exceedingly high activity against erythromycin-susceptible gram-positive cocci, Mycoplasma pneumoniae, and Legionella pneumophila makes it and important alternative choice in the therapy of respiratory tract infections. Also, it has shown high activity against Chlamydia trachomatis, and high urinary clearance of this unmetabolized molecule, important properties which would render it a special role in the treatment of genitourinary tract infections. Azithromycin, a 15-membered derivative has shown enhanced basicity (due to the nitrogen atom in its lactone ring), longer half-life and lower protein bindings. Its exceptional activity against Hemophilus influenzae, Branhamella catarrhalis, Neisseria gonorrhoeae, Ureaplasma urealyticum and gram-negative bacteria, and its high concentration in tonsillar, pulmonary, prostatic and female reproductive tract tissues, assigns it an honorific place among the macrolides in the therapy against respiratory tract and genitourinary tract infections. Its role against T. gondii deserves further study, but points out this agent as a promise against this parasite.


Disseminated histoplasmosis.

We have presented the microbiology, pathogenesis, clinical presentation, diagnosis and treatment of disseminated histoplasmosis. Emphasis is presented on the manifestations in the HIV infected patient.


Infected bullae: a recognizable entity.

Infected bullae are seldom diagnosed. They are frequently confused with pulmonary abscess. We present a case with a characteristic evolution. Bullae were identified previous to the infection. The illness developed gradually with scanty sputum, pleuritic pain and finally a febrile illness. Fever abated 48 hours after initiating two weeks of therapy with intravenous Clindamycin and Amikacin. As it is commonly seen, radiographic resolution was slow but it was complete in 8 weeks. Recognition of infected bullae is important to avoid inappropriate diagnostic or therapeutic interventions.


Synthesis, hypoglycemic and toxicological evaluation of new 1,2-benzisothiazolyl derivatives.

The synthesis of 1,2-benzisothiazol-3-ylguanidines, 1,2-benzisothiazol-3-ylbenzensulphonylureas and 1,2-benzisothiazol-3-ylbenzensulphonamides is described. Some of the new compounds showed moderate hypoglycemic activity but most of them caused serious acute toxic effects.


Controlled release of proteins and peptides from hydrogels synthesized by gamma ray-induced polymerization.

Hydrogels prepared by radiation-induced polymerization at a low temperature have been used as carriers for the controlled release of peptides and proteins. It was found that polymerization of 2-hydroxyethyl methacrylate in the presence of poly (ethylene glycol) methyl ether (MPEG) enabled the more porous and swellable matrics to be obtained, the higher the molecular weight of MPEG. As a consequence, protein release took place at an increasing extent and, provided that MPEG molecular weight was high enough, high molecular weight proteins could also be released. Such a state of affairs was not met in the case of hydrogels based on poly (2-hydroxyethyl acrylate). SEM analysis revealed that even high molecular weight MPEG did not give rise to any porosity, even though the degree of swelling was very high. As a result, no protein release was observed. It was therefore concluded that control of hydrogel porosity for the controlled release of large proteins is of overwhelming importance.


Synthesis and evaluation of gastroprotective and antiulcer activity of some 2-substituted-1H-imidazo[4,5-b] pyridines and -1H-benzimidazoles.

Several compounds possessing imidazo[4,5-b]pyridine and benzimidazole structure bearing substituents in position 2 were prepared in order to evaluate an anti-ulcer and gastroprotective activity in rat pylorus ligature, in comparison with omeprazole. Among sixteen compounds taken as representatives of the synthetised series only one (3d) showed a good activity by i.m. administration at 50 mg/kg, while by oral administration of 100 mg/kg a certain number was active and in some cases this activity was quite superior to that of omeprazole.


Synthesis, spectral studies and biological activities of some N-bridged heterocycles derived from 3-arylaminomethyl-4-amino-5-mercapto-1,2,4-triazoles.

Synthesis of four 3-arylaminomethyl-4-amino-5-mercapto-1,2,4-triazoles starting from substituted anilines is described. These triazoles were employed in the synthesis of some N-bridged heterocycles carrying arylaminomethyl substituents. All the newly synthesized compounds were characterized by analytical, NMR and mass spectral studies. Some of the newly synthesized compounds were screened for their antibacterial and antiviral properties.


Synthesis of some new fluorine bearing trisubstituted 3-thioxo-1,2,4-triazin-5-ones as potential anticancer agents.

In a search for new anticancer agents fluorine bearing trisubstituted 3-thioxo-1,2,4-triazin-5-ones (2-12) have been prepared and characterized by their elemental analysis, UV, IR and 1H-NMR spectral data. The in vitro anticancer activity of all the compounds has been determined. Compounds 3 and 7 showed a moderate activity against Leukemia/Lymphoma, Small/Non small Cell Lung, Colon carcinoma and Melanoma Cells.


Synthesis and local anesthetic activity of some derivatives of N,N-dimethyl-2-(2-alkoxyphenylcarbamoyloxy)-1,1-dimethylethyl-ammonium chlorides.

A series of 8 new derivatives of 2-alkoxyphenylcarbamoic acid were synthesized and assayed for local anesthetic activity. The above compounds were isolated as hydrochlorides and their structure was proved by 1H NMR, 13C NMR and IR spectroscopy. The index of anesthetic activity of the compounds in infiltration and surface anesthesia increases with the length of the alkyl chain and, except for the C1-C3 congeners, all higher homologues proved to be significantly more active than the standard reference compounds, procaine and cocaine, respectively. Toxicity of the drugs decreases with increasing the chain length and is within acceptable limits.


Structure-activity studies on a 1,2,3-triazole derivative, a potent in vitro inhibitor of prostaglandin synthesis: the role of the heterocyclic ring.

This paper reports further structural modifications concerning the 1,2,3-triazole ring of the compound A, an effective in vitro inhibitor of prostaglandin synthesis. The introduction of different heterocyclic rings provided further information about of the role of the heterocyclic ring in enzymatic inhibition, as regards the number and position of the nitrogen atoms, the electronic effects, basicity, steric hindrance and hydrophilicity. The benzimidazole derivative 4e proved to possess a high enzymatic inhibitory activity.


Cyclohepta[b]pyran derivatives with platelet antiaggregating and other activities.

The synthesis of some N,N-disubstituted 4-amino-6,7,8,9-tetrahydro-3-phenylcyclohepta[b]pyran-2(5H)-ones by reaction of phenylchloroketene with a series of N,N-disubstituted 2-aminomethylenecycloheptanones, followed by dehydrochlorination of the primary adducts with DBN, is described. Some compounds showed a platelet antiaggregating activity in vitro superior or comparable to that of acetylsalicylic acid, as well as weak local anesthetic, antiinflammatory and analgesic activities in mice and rats.


1-Phenyl-1H-indazole derivatives with analgesic and anti-inflammatory activities.

The synthesis of 4-hydroxy-1-phenyl-1H-indazole-5-acetic acid 5 4-methoxy-1-phenyl-1H-indazole-5-yl-acetic acid 7 and 5-benzyl-1-phenyl-1H-indazol-4-ol 8, starting from 1,5,6,7-tetrahydro-1-phenyl-4H-indazol-4-one, is described. These compounds showed in mice an analgesic activity superior to that of acetylsalicylic acid and comparable to that of dipyrone; moreover, compound 5 exhibited an appreciable anti-inflammatory activity in rats. Grossbehavioral effects and acute toxicity in mice are also reported.


Cannabinoid acids analysis.

The cannabinoid pattern of vegetable preparations from Cannabis sativa (hashish, marijuana) allows to recognize the phenotype of the plants, to be used as drug or for fiber. Cannabinoid determination by analytical point of view has represented some problems caused by the complex composition of the hexane extract. Capillary gas chromatography of the hexane extracts of vegetable samples, shows the presence of rather polar constituents that eluted, with noticeable interactions, only on polar phase. The compounds can be methylated by diazomethane and silanized (TMS) by silylating reagents. The methyl and methyl-TMS derivatives are analyzed by high resolution gas chromatography (HRGC) and by gas chromatography-mass spectrometry (GC-MS). The identification of the compounds shows their nature of cannabinoid acids, which the main by quantitative point of view results the cannabidiolic acid (CBDA). It is known that the cannabinoid acids are thermally unstable and are transformed in the corresponding cannabinoids by decarboxilation. This is of interest in forensic analysis with the aim to establish the total amount of THC in the Cannabis preparations, as the active component.


Red blood cell partitioning of gallopamil, verapamil and norverapamil.

In-vitro binding of calcium-antagonists gallopamil and verapamil (and its main metabolite norverapamil) to human red blood cells (RBCs) was investigated. The drugs are bound reversibly and dose dependent to RBCs in the same order of magnitude, with partition-coefficients of kRBC = 0.12-0.34 for gallopamil, kRBC = 0.10-0.30 for verapamil and kRBC = 0.10-0.27 for norverapamil. The data indicate that, although RBCs may act as subcompartments of the blood for this class of compounds, they may have no influence on therapeutic plasma concentrations, due to their low kRBC.


Spectrophotometric determination of diltiazem in dosage forms.

A simple and reproducible method for determination of diltiazem in bulk and in dosage forms is presented. The method is based on formation of hydroxamic acid, which reacts with iron (III), forming a complex with a maximum absorption at 525 nm. Assay procedure for diltiazem dosage form requires thin-layer chromatographic separation prior to colorimetric analysis.


[Detection of an inhibition of benzo(a)pyrene and sodium azide induced mutagenesis by extracts from human feces].

Low levels of mutagenic activities were detected in only 5-25% of the feces of people on a normal mixed-western diet, when feces were extracted by solvents and extracts were analyzed for mutagenicity with Ames' standard Salmonella/microsome assay. Since mutagens are know to be present in this type of diet and may be synthesized endogenously by bacteria in the large bowel, the question is if antimutagenic compounds mask the presence of genotoxic substances. We therefore tested the inhibition of known mutagens--benzo(a)-pyrene and sodium azide--by acetone/ethyl-acetate extracts of lyophilized feces in model experiments. These extracts completely suppressed the mutagenicity of benzo(a)pyrene and reduced the mutagenicity of sodium azide by about 60%, but were non-cytotoxic. Using gel filtration over Sephadex LH-20 and subsequent silica gel column chromatography we found that the inhibitors were polar organic compounds with molecular weights about 500 or more. The inhibitory effect could not be changed by esterification or saponification methods but was completely abolished by alkaline potassium permanganate oxidation. We therefore conclude that the antimutagenic activities might be identical with bile pigments.


[The behavior of fiber-reinforced plastics during laser cutting].

The pattern of the organic emissions, which are produced by processing of fibre reinforced plastics (epoxy resins reinforced by aramid and glass fibres and phenol resins reinforced by aramid fibre) with laser beam was studied and the concentrations of the main components determined. Despite the application of plastic materials with different chemical structures, the observed patterns are very similar. Mainly aromatic hydrocarbons are emitted, especially benzene and toluene, as well as some heteroatom-containing aromatic hydrocarbons (e.g. phenol). By use of oxygen as process gas the emissions during cutting of glass fibre reinforced plastics can be reduced, while they will be constantly high with aramid fibre reinforced plastics.


Influence of the pH-value on the growth of Staphylococcus epidermidis, Staphylococcus aureus and Propionibacterium acnes in continuous culture.

A cutaneous isolate of Staphylococcus epidermidis, Staphylococcus aureus and Propionibacterium acnes was grown in continuous culture at varying pH-values ranging from 5.0 to 8.5. In terms of the specific growth rate as well as the bacterial density during the plateau phase there were remarkable differences. In particular, Propionibacterium acnes grew much better in the pH 6.0 to 7.0 range than in a more acidic or alkaline milieu. Staphylococcus epidermidis resembled Staphylococcus aureus showing no major difference at pH 5.5 and 7.0. These findings substantiate the hypothesis that minor changes of the pH in the pH 5.5 to pH 6.0 range as to be induced by chemically neutral or alkaline skin cleansers on the human skin surface can increase the number of propionibacteria but not staphylococci remarkably due to the relative alkalinity by itself.


[Halogenated organic compounds in swimming pool water].

18 different swimming pool waters in the Bonn-Rhein-Sieg area were analysed for trihalogenmethanes, halogenated acetic acids, 2,2-dichloropropionic acid, dihalogenacetonitriles and chloral hydrate. Most substances mentioned above were determined in almost every sample reaching total concentrations up to 538 micrograms/l. Considering the results one can see that some of the main pollutants are polar organic compounds like di- and trichloroacetic acid as well as chloral hydrate.


T cell activation and antigen presentation in human thyroid autoimmunity.

In 1983, a hypothesis concerning the relevance of class II expression and antigen presentation to the induction and maintenance of endocrine autoimmunity was published. This article reviews the evidence that has been marshalled to support this concept, both in man, chiefly in Graves' disease, and in murine systems. New data concerning the multiplicity of thyroid autoantigens recognized by in vivo activated thyroid infiltrating T cells are compatible with this concept since the thyroid epithelial cells are the source of these antigens. Whether other autoimmune diseases also employ the same mechanism remains to be ascertained, as does the initial trigger for the autoimmune process.


Studies on the role of tumor necrosis factor in murine and human autoimmunity.

We have analyzed the roles of tumor necrosis factor alpha (TNF-alpha) in human systemic lupus erythematosus (SLE) and murine models of lupus as well as in type 1 diabetes in NOD mice. These studies suggest an important role for TNF-alpha in the pathogenesis of autoimmune disease. Rather than being involved mainly in the effector arm of the inflammatory process of autoimmune organ destruction, our data suggest a primary involvement in some of the basic mechanisms of the autoimmune process. Evidence has been presented that emphasizes the possibility of the involvement of this cytokine in the genetic predisposition to SLE. The data may imply that the effect of TNF on the immune system may be more relevant to the pathogenesis of the autoimmune disease than direct local effects at some target organs. Based on the data presented, one should be cautious in extrapolating the effects of this cytokine in various in vitro systems to the in vivo situation.


The effects of cyclosporin A on non-T cell components of the immune system.

Cyclosporin A (CsA) is a potent inhibitor of cytokine (IL-2-IL-6, IFN gamma) production by CD4+ T lymphocytes stimulated via the T cell antigen receptor pathway. This action results in indirect inhibitory effects on the growth and differentiation of B lymphocytes (IL-4 and IL-6). Using experimental models, it has also been shown that the functional activities of mononuclear phagocytes (IFN-gamma) and other antigen-presenting cells, production of mast cells (IL-3) and eosinophils (IL-5) and the activity of natural killer (NK) cells may be inhibited indirectly by CsA. In addition, however, CsA blocks B cell responses to Ca(2+)-dependent signals (e.g., anti-IgM) downstream of phosphatidyl inositol diphosphate hydrolysis; Ca(2+)-independent responses (e.g., to LPS or IL-4) are largely unaffected. In general terms, the functions of macrophages are unchanged or reduced in the presence of CsA. These include phagocytic activity in vitro and in vivo, chemotactic migration, superoxide and H2O2 production, protein (including monokine) secretion and MHC gene product expression. Antigen presentation (e.g., by epidermal Langerhans cells) may be affected, especially at high drug concentrations. There is recent evidence that CsA inhibits mediator (histamine and prostaglandin) release from human mast cells and that mucosal mast cell numbers may be diminished in CsA-treated animals exhibiting graft-versus-host disease or helminth infections.


Cyclosporin monitoring: its role in autoimmune indications.

This paper describes some of the methodological problems related to the measurement of cyclosporin. The clinical value of the measurements following organ transplantation are discussed and those areas also applying to autoimmune indications are highlighted. It is concluded that the routine use of cyclosporin monitoring in samples from patients receiving the drug for autoimmune indications is unlikely to be of significant value as a guide to efficacy or toxicity. However, some settings, such as suspected poor patient compliance, for the avoidance of potential drug interactions and for research on the absorption of new formulations are considered to be useful applications of the methodology.


The optimal use of cyclosporin A in autoimmune diseases.

The optimal use of cyclosporin (CsA) in autoimmune diseases aims at achieving the best risk/benefit ratio and ensuring the absence of potentially irreversible adverse effects, particularly with respect to the kidney [corrected]. The experience gained with CsA therapy in more than 3,000 patients with autoimmune diseases is the basis for the current recommendations: the initial dose should be the lowest effective one and not exceed 5 mg/kg/day in non-life-threatening conditions; treatment should be as brief as possible (2-4 months) in cases of inefficacy; once a satisfactory clinical improvement has been achieved, the treatment should be maintained in the long-term using the lowest individually titrated effective dose; the dose of CsA should be decreased when serum creatinine rises by more than 30% above pre-CsA level. Continuous clinical and biological monitoring (especially of blood pressure and serum creatinine) is mandatory as long as CsA is prescribed. When these conditions are fulfilled, CsA may be an effective and safe therapy for selected autoimmune diseases, even in long-term treatment.


Timing of immunosuppression in the natural history of autoimmune diseases.

We have insufficient data to guide us to the optimal timing of immunosuppression in the natural history of any autoimmune disease. Moreover, there are differences among the many autoimmune diseases and the many drugs available for use. Nevertheless, certain principles have emerged. Prophylactic non-specific immunosuppression prior to the onset of the immune-mediated process often is of minimal benefit. Vigorous immunosuppression shortly after the onset of the immune-mediated process is most effective; many agents are of benefit at such times. If the disease has progressed to substantial clinical involvement, certain drugs previously useful may no longer be effective. At such a time of moderately advanced clinical involvement, only selected agents may suppress the disease. With substantial loss of function of irreplaceable organs, or parts thereof, immunosuppression becomes progressively less effective. Such drugs can interfere with inflammatory processes, but are of little benefit after deletion of cells or scarring of an organ. Therefore, to have any benefit, immunosuppression must be instituted prior to the time of irreversible loss of critical organs or parts thereof.


The case for early intervention in rheumatoid arthritis.

A case for early intervention in rheumatoid arthritis is presented on the basis of the following observations: (1) established rheumatoid arthritis of 5 years duration or more is a progressive disease in most patients; (2) most patients with rheumatoid arthritis already have evidence of permanent radiographic damage within the first 2 years of disease; (3) the morbidity and mortality of rheumatoid arthritis are predicted by baseline data indicating more severe clinical status, rather than drug toxicity or causes 'unrelated' to rheumatoid arthritis; (4) currently used therapies, even those documented to be effective in randomized controlled trials, are not effective in most patients over periods of 2 years or longer. It is not known whether earlier intervention will result in better outcomes. However, these phenomena suggest a need for studies to evaluate early aggressive intervention in rheumatoid arthritis.


Immunologically specific treatment of spontaneous insulin-dependent diabetes mellitus of NOD mice.

This paper discusses the possibility of immunologically specific therapy of spontaneous autoimmune disease. The results of animal studies encourage optimism.


The expanding use of immunosuppression in the treatment of non-infectious ocular disease.

Cyclosporin has been used for an increasing number of ocular indications. This review attempts to put into perspective the present scope of cyclosporin use in the treatment of ophthalmic disorders. This includes a review of randomized masked trials performed to date and work done to minimize and characterize cyclosporin induced renal toxicity. In addition, the ocular surface is an ideal target for topical cyclosporin therapy, and the status of this therapeutic approach is reviewed.


Insulin autoimmunity: the rate limiting factor in pre-type I diabetes.

Type I diabetes is preceded by a series of autoantibodies including recently recognized subtypes of cytoplasmic islet cell antibodies (ICA) which we have termed 'restricted' and 'non-restricted'. Amongst the autoantibodies detected in prediabetics the antibodies to insulin are unique in that their levels correlate with the rate of progression to type I diabetes. The levels of insulin autoantibodies appear to be stably regulated prior to the appearance of ICA and the highest levels are associated with expression of DR4. The above studies have led to the hypothesis that an immune response to insulin is an early and central feature of anti-islet autoimmunity and that such as immune response when associated with loss of tolerance to other islet antigens (e.g. ICA) is pathogenic.


Low-dose cyclosporin therapy of ocular inflammation: preliminary report of a long-term follow-up study.

Cyclosporin A (CsA) is an effective therapy for severe intraocular inflammation but nephrotoxicity and hypertension are major side effects even in low dose in combination with oral corticosteroids and clinical studies on the long-term effects of low-dose CsA therapy outside the field of organ transplantation are lacking. This multicentre, open, longitudinal study has been established to evaluate the long-term efficacy and side effects of low-dose CsA therapy (initial dose less than or equal to 5 mg/kg/day, with a maximum dose of 7 mg/kg/day, and total treatment duration greater than 3 months) in severe ocular inflammation where conventional therapy had failed to control the disease or caused intolerable side effects. Visual response to treatment, clinical signs and symptoms of side effects, biochemical and haematological parameters have been recorded at 3-monthly intervals since January 1987 and will continue until December 1993. Data for 74 patients (age 35.5 +/- 16.6 years) and 293 follow up visits are presented in this preliminary report. [table: see text] Other side effects include (% of all visits): hypertrichosis (4.2), headache (2.8), cramps (1.8), arthropathy (1.8), paraesthesiae (1.8), abdominal pain (1.5), weakness (1.5), dyspepsia (1.4), nausea (1.4), others (4).


Cyclosporin A in asthma therapy: a pharmacological rationale.

Lymphocytes play a central role in immunological reactions and control the mobilisation and recruitment into lung tissue of eosinophils and mast cells. Since cyclosporin A can profoundly influence lymphocyte activation, it is appropriate to consider this drug as a novel anti-asthma therapy. Inhalation of low doses of cyclosporin A strongly inhibits the influx of inflammatory cells into the airways during acute allergic reactions, even though associated bronchospasm and airway hyper-reactivity are undiminished. It is suggested that cyclosporin A will be an effective anti-asthma therapy with an anti-asthma profile resembling that of established glucocorticosteroids.


Post-thymic tolerance to self antigens.

There is now convincing evidence for the imposition of self-tolerance by means of the clonal deletion of self-reactive T cells operating within the thymus. Since not all self components may be encountered there, the question must be asked whether tolerance can occur post-thymically. To test this, we have used transgenic technology to direct expression of a known 'non-self' gene, H-2Kb, to the insulin producing beta cells of the pancreas of mice. H-2Kb-bearing skin, but not skin from other mouse strains, failed to be rejected by the 'RIP-Kb' transgenic mice indicating specific tolerance. Following in vitro stimulation, their spleen cells could not kill H-2Kb-bearing targets, but could respond to third party targets. Their reactivity to H-2Kb was restored by providing them with IL-2. Two hypotheses could account for the above: tolerance results either from the deletion or functional silencing of high affinity effector cytotoxic cells or of regulatory, IL-2-producing helper T cells. Since it is difficult to distinguish between these, we have produced a second series of transgenic mice with rearranged T cell receptor (TCR) genes encoding an anti-H-2Kb TCR, and obtained 'double transgenic' offspring by mating these mice with RIP-Kb mice. The TCR utilized the V beta 11 segment which can be detected by a monoclonal antibody. Although the double transgenic mice were tolerant of H-2Kb, there was no evidence of deletion of anti-H-2Kb T cells. It seems, therefore, that a non-deletional mechanism operates to induce post-thymic tolerance.(ABSTRACT TRUNCATED AT 250 WORDS)


A multicenter randomized study comparing cyclosporin-A alone and antithymocyte globulin with prednisone for treatment of severe aplastic anemia. The cooperative group on the treatment of aplastic anemia.

This article reports the results of a randomized multicentric study comparing the efficacy of antithymocyte globulin (ATG) with cyclosporin-A (CsA) as first line therapy for severe aplastic anemia (SAA). Patients were randomized to receive ATG and prednisone (PDN) or CsA; hematological response and toxicity were compared. At 3 months, patients who had no or minimal response received the alternative therapy in order to assess the value of a sequential immunosuppressive therapy for treatment of severe aplastic anemia. One hundred and sixteen patients were analysed, 60 received CsA and 56 received ATG/PDN. The actuarial survival was 55% with a median follow-up time of 19 months. There was no significant difference in survival between the two groups. The main prognostic factor was the absolute neutrophil count (ANC) at entry: patients with ANC less than 0.2 x 10(9)/L had a significantly lower survival when compared with patients with an ANC greater than 0.2 x 10(9)/L (P = 0.0001). At 3 months, 16% of patients had a complete or partial response and a cross-over treatment was given to 68 patients. At 12 months, 77 patients were alive, with a complete or partial response in 47 patients. Patients who had responded to the first treatment had a better recovery of bone marrow failure than those who had a sequential immunosuppression. The main complication was infection which was more often observed and was more often lethal during ATG and PDN. This study demonstrates that CsA is comparable to ATG for primary treatment of SAA, but it is less toxic with fewer infectious deaths.(ABSTRACT TRUNCATED AT 250 WORDS)


Psoriasis: immunopathology and long-term treatment with cyclosporin.

Psoriasis is a disorder of abnormal keratinocyte proliferation mediated by T lymphocytes. Initiation of psoriasis is associated with an influx and activation of CD4 T lymphocytes into the dermis and epidermis, and resolution with an influx and activation of CD8 T lymphocytes within the epidermis. As a result of these observations it was postulated that cyclosporin (CsA) should prove to be an effective treatment for psoriasis. A total of 82 patients with psoriasis have been treated with CsA. Thirty patients with moderate disease were treated for periods up to 12 weeks, and comprised a short-term study. Two groups have been treated with long-term CsA. A group of 18 patients have been treated for greater than 1 year but less than 3 years, (mean 2.1) and a group of 10 patients for 4.0-5.5 (mean 5) years. A total of 15% of patients can be maintained on less than or equal to 2 mg/kg/day; 55% on 3 mg/kg/day, 80% on 4 mg/kg/day, and 92% on 5 mg/kg/day. Hypertension occurred in 17% of patients in the short-term study, 29% in the 2.1-year group and 44% in 5-year group. Blood pressure returned to normal in all hypertensive patients when CsA was discontinued for 1 month. Nephrotoxicity was assessed by serum creatinine levels and glomerular filtration rate (GFR) and renal biopsy in the 5-year group. In the short-term group there was no significant rise in the serum creatinine.(ABSTRACT TRUNCATED AT 250 WORDS)


Cyclosporin for palmoplantar pustulosis.

Palmoplantar pustulosis (PPP) is a chronic skin disease characterized by persistent erythematous, scaly plaques incorporating sterile pustules on palms and soles, which is resistant to most treatments. Recently, two published uncontrolled studies suggested that cyclosporin (CsA) could be an effective treatment for PPP. Similarly, an unpublished, randomized, placebo-controlled study showed that CsA is effective in preventing new pustule formation in PPP. In the present paper we review the treatment of PPP with special emphasis on CsA treatment.


Extension of indications of immunosuppression in inflammatory bowel disease.

Treatment of the two chronic inflammatory bowel diseases (IBD), Crohn's disease (CD) and ulcerative colitis (UC) is still hampered, because we do not understand the pathophysiological processes behind them. Are they caused by a normal inflammatory response reacting towards a foreign agent, or are they caused by an abnormal inflammatory response against a common or even universal agent? Our only choices at present are symptomatic treatments, including immunosuppressive drugs. To make the necessary controlled testing of new immunosuppressive drugs ethically acceptable, we have to choose carefully between different designs: comparing a new drug against placebo in overall treatment failures, to add the new drug or placebo to ongoing treatment (addition design), or to apply a classical group comparison between test drug and placebo in moderately active cases. There exists a strong need for extension of indications for immunosuppressive and anti-inflammatory treatments: to be able to treat long-term instead of short-term, to do without extensive surgery in some UC cases, to treat severe UC cases with isolated distal involvement, to treat CD patients with short bowel syndrome, and to treat CD patients with fistulae and severe extra-intestinal manifestations. Future drug treatment of IBD will probably still be based on immunosuppressive and anti-inflammatory drugs, while waiting for a pathophysiological breakthrough in these two diseases.


Immunointervention in primary biliary cirrhosis: an overview.

Since autoimmune processes are probably involved in the early stages of primary biliary cirrhosis (PBC), immunomodulatory drugs have been investigated with the aim of prolonging survival, delaying transplantation, slowing histological progression and relieving symptoms. Corticosteroids, azathioprine, chlorambucil and, more recently, cyclosporin A and methotrexate have all be subjected to clinical investigation. In the latest of these, a European multicentre trial, cyclosporin A has been shown to delay death or transplantation with a reduction in liver related deaths and slowing of the rise of serum bilirubin. The incidence of nephrotoxicity and hypertension are low at the doses used.


Immunointervention with cyclosporin A in autoimmune neurological disorders.

Cyclosporin A has proven a remarkably effective compound in suppressing disease activity in a number of animal models of autoimmune neurological disorders. During the last decade a number of controlled clinical trials have been carried out in human neurological disorders which are considered to be immune-mediated. The results of those trials are reviewed.


Treatment of the nephrotic syndrome with cyclosporin A.

The three main causes of primary nephrotic syndrome are minimal change nephropathy, focal glomerulosclerosis and membranous nephropathy. Corticosteroids obtain remission of proteinuria in most patients with minimal change nephropathy. Many patients, however, show either frequent relapses or steroid dependency. A short course of cyclophosphamide or chlorambucil can achieve stable remission in many of these patients but alkylating agents cannot be repeated, their toxicity being cumulative. Analyses of the available studies showed that some 80% of patients can be maintained in remission with cyclosporin A (CsA) but relapse occurs when the drug is stopped. Severe side effects are rare. In particular, the mean values of creatinine clearance did not deteriorate in cyclosporin-treated patients and repeat renal biopsy showed only mild changes in some patients. There is no definite treatment for focal glomerulosclerosis. An analysis of 10 clinical trials showed that some 17% of nephrotic patients may enter complete remission of proteinuria and another 13% may attain partial remission of the nephrotic syndrome with CsA. There is concern over the use of this drug since cases of irreversible renal dysfunction have been reported. However, retrospective reviews of the available studies showed that the mean serum creatinine levels did not modify if patients had normal renal function when given CsA. A 6-month course of methylprednisolone and chlorambucil may obtain remission of the nephrotic syndrome in some 60% of patients with membranous nephropathy. Some trials have shown that CsA may improve proteinuria and there is also some suggestion that the drug might protect against renal function deterioration. Thus, when given at correct doses, CsA may exert an anti-proteinuric effect without deteriorating renal function, suggesting that the drug may represent a further tool in treating the primary nephrotic syndrome.


Pharmacokinetic strategies for cyclosporin therapy in organ transplantation.

Marked interindividual variations in cyclosporin (CsA) produce disparate clinical results in organ transplant recipients. In an attempt to eliminate marked deviations of insufficient or excessive CsA concentrations consequent to the administration of uniform drug doses, test dose pharmacokinetics were performed on each potential organ transplant candidate. An intravenous 3 mg/kg test dose delivered over 3 h proved to be readily performed, namely 53% perfect studies, and relatively reliable, namely 73% of observed concentrations within 10% of the predicted values. Furthermore, the use of CsA doses predicted by pretransplant studies reduces the incidence of delayed graft function, early rejection episodes and transplant loss. The oral test dose study predicted a suitable amount of CsA to achieve sufficient gastrointestinal absorption but was less accurate than the iv prediction method: namely, 40% of observed post-transplant concentrations were within 10% of the predicted target value. Furthermore, patients who received oral doses predicted by the test dose strategy showed no improvement in the incidence of acute rejection episodes between 7 and 60 days, and only modestly improved serum creatinine values. The lower accuracy of predictions from oral test dose studies may reflect the impact of non-linear oral (as opposed to iv) drug pharmacokinetics, of variable diet, and/or of altered postoperative gastrointestinal function.


Immunological monitoring in organ transplantation and autoimmune disease.

The clinical manifestations of graft rejection or autoimmune disease represent the terminal phase in a complex sequence of inflammatory events. Although the spectrum of response to injury within each tissue is normally narrowly defined, the immune mechanisms involved may be heterogeneous, influenced by the immune status of the host, the nature of the stimulus (i.e. viral antigen, allostimulation, or aberrant recognition of self), and the use of exogenous immunosuppression. Within this framework, immunological monitoring is employed to distinguish the causative effector mechanisms, to characterize the disease course, to tailor therapeutic intervention, and to monitor treatment impact in individual immunological disorders.


The effect of cyclosporin on renal function.

The depression of renal function caused by cyclosporin does not generally reflect permanent kidney damage but is caused by a reversible vasoconstriction, with no relevant changes in tubular function. Serum creatinine may remain within the normal range during therapy, but any decline in renal function can be detected by a rise in serum creatinine above the baseline value. Measurements of glomerular filtration rate before and during therapy have shown the degree of renal dysfunction in individual patients to correspond to their rise in serum creatinine. The cause of renal vasoconstriction is uncertain but animal experiments have highlighted several possibilities. These include: (1) albumin leakage with circulatory volume contraction; (2) enhanced Ca mobilization in contractile cells; (3) activation of a renin-like enzyme in vessel walls; and (4) renin accumulation in the renin-producing cells of the afferent arteriole. Such mechanisms, although seeming to operate at different doses, may act in unison at high doses, when renal function is most severely depressed.


Serial kidney biopsies in patients with nephrotic syndrome treated with cyclosporin.

A total of 21 patients with severe steroid-resistant or steroid-dependent nephrotic syndrome have been treated with cyclosporin (CsA) over a period of 6 to 71 months. A permanent treatment was started in patients with complete [c] (proteinuria less than 0.3 g/day) or partial [p] (0.3 to less than 3.0 g/day) remission. In four cases proteinuria was markedly reduced but still greater than 3 g/day. Informed consent was obtained and all patients agreed to further control biopsies. Out of 14 patients treated for longer than 6 months, a fall in creatinine clearance was seen in three cases, including two patients with focal sclerosis who required dialysis treatment. Control biopsies were performed once in 11 and twice in seven patients. In two cases definite CsA-related damage was diagnosed after 1 and 24 months of treatment. Possible CsA-induced damage could not be excluded in three additional cases. In all other biopsies no CsA-related alterations could be demonstrated up to 47 months of treatment. The mean values of the creatinine clearance showed no deterioration. The drop off in a few cases has been related to the basic disease.


Malignancy in autoimmune diseases.

Rheumatoid arthritis (RA) represents the autoimmune disease that has been most studied in relation to malignancy. An examination of all published cohort studies has indicated a 9.7-fold increase of non-Hodgkin's lymphoma among RA patients after immunosuppressive therapy, and a 2.5-fold increase in the absence of such treatment. Corresponding data for Sjögren's syndrome point to a similar contrast. These findings are inseparable from the hypothesis of impaired immunosurveillance which implies that malignancy is promoted by defects in the immune system. Studies of individuals treated with immunosuppressive drugs, particularly to prevent graft rejection, have indicated that immunosurveillance operates only against a restricted range of neoplasms. These include non-Hodgkin's lymphoma (NHL), squamous cell skin cancer, Kaposi's sarcoma and cervical carcinoma. Other states of immune impairment including AIDS are also associated with marked increases of NHL. There is a striking correspondence between malignancies for which there is epidemiological or laboratory evidence for a virus aetiology and those that are increased by immune impairment. In this respect the epidemiological evidence accords with experimental work that immunosurveillance primarily operates against neoplasms of viral origin. It is therefore possible that a viral aetiology also underlies the excess of NHL in certain autoimmune disorders, particularly after immunosuppressive therapy.


Specific and non-specific autoreactive immunity.

Most autoimmune diseases are HLA-associated which supports the notion that they are dependent upon specific immune activation of a limited set of T cell clones. Findings which imply that induction of autoimmune reactivity probably does not differ from normal immune responses are discussed. The possibility of transferring autoimmune disease using T cell clones indicates that target structures for auto-immune attack are also present in healthy individuals. In the present article, it is argued that autoimmune reactions and immunity against nominal conventional antigens in principle are effected and regulated by similar mechanisms. It is assumed that persistent tissue damage occurs if immune attack is directed against tissues that cannot be regenerated, such as in diabetes, or are only slowly reconstituted, such as in rheumatoid arthritis. Normal immune responses are regulated by various inflammatory mediators and cytokines/interleukins. The joint of patients with rheumatoid arthritis is discussed as a model for propagation of immune reactions and tissue destruction in autoimmune disease. Of the different cytokines which are present in the synovial fluid or produced by cells in the synovial tissue, most are presumed to have originated in macrophages/monocytes such as IL-1, IL-6, IL-8, TNF-alpha and TGF-beta. Even so, T cells are believed to have an important role for the continued reactivity associated with autoimmune disease. This discrepancy can be explained in different ways. T cell products might escape detection because they are short-lived, they are immediately consumed or they are produced only during short time intervals.(ABSTRACT TRUNCATED AT 250 WORDS)


Generation of mutant mice lacking surface expression of CD4 or CD8 by gene targeting.

With the use of homologous recombination in embryonic stem cells, two new strains of mice lacking surface CD4 or CD8 expression have been generated. It is hoped that they will be useful mouse strains for the study of autoimmune diseases, tissue transplantation rejections and tumour rejections.


Inhibition of T cell activation by MHC blockade: a possible strategy for immunointervention in autoimmune diseases.

Autoimmune diseases result from the activation of self-reactive T cells induced by autoantigens or by foreign antigens cross-reactive with an autoantigen. A striking characteristic of autoimmune diseases is the increased frequency of certain HLA alleles in affected individuals. Moreover, as demonstrated for example in rheumatoid arthritis and insulin-dependent diabetes mellitus, class II alleles positively associated with autoimmune diseases share amino acid residues in the hypervariable HLA regions involved in peptide binding. Therefore, it is likely that disease-associated HLA class II molecules have the capacity to bind the autoantigen and present it to T cells, thereby inducing and maintaining, under appropriate conditions, the autoimmune disease. The data reviewed here demonstrate MHC-selective inhibition of antigen-induced T cell responses in vivo by parenterally administered soluble, MHC-binding peptide competitors, under conditions in which the competitor is not immunogenic. This suggests the feasibility of a therapeutic approach based on blockade of MHC class II molecules in the treatment of HLA-linked autoimmune diseases.


T cell receptor peptide therapy for autoimmune disease.

Synthetic peptides corresponding to germline T cell receptor (TCR) V beta sequences shared by encephalitogenic T cells can prevent and treat experimental autoimmune encephalomyelitis in rats. The operative mechanism apparently involves boosting of anti-TCR immunity that develops during the course of experimental autoimmune encephalomyelitis (EAE), leading to the induction of autoregulatory T cells and antibodies. Striking parallels are present between patients with multiple sclerosis and animals with EAE in the T cell frequency and TCR V gene bias of BP reactive T cells, suggesting the involvement of an encephalitogenic process in multiple sclerosis. Preliminary trials with the appropriate human TCR peptides indicate that anti-TCR immunity can be boosted efficiently and safely, with concomitant loss of BP response, thus providing an effective strategy for selective regulation of autoimmunity in man.


Resistance of adult keratinocytes to differentiation-induced decrease in Ha-ras mRNA levels observed in newborn keratinocytes.

During two-stage mouse skin tumorigenesis, the mouse c-Ha-ras oncogene undergoes activation by point mutation after initiation with polycyclic aromatic hydrocarbons. Furthermore, initiated epidermal cells containing an activated Ha-ras oncogene have been shown to be resistant to calcium-induced terminal differentiation. However, the relationship between Ha-ras expression and the differentiation process is not well understood in either normal or initiated cells. Before attempting to explore the role of Ha-ras expression in epidermal differentiation during tumorigenesis, we felt that investigation of Ha-ras gene expression in normal primary epidermal cells undergoing differentiation was warranted, since primary cultures of normal newborn and adult keratinocytes presumably contain the stem cells from which skin tumors arise. In the present studies, northern blot analysis was used to compare Ha-ras expression in normal newborn and adult epidermal cells undergoing differentiation. Steady-state levels of Ha-ras mRNA remained unchanged in primary cultures of normal adult epidermal cells during calcium-induced differentiation, whereas steady-state levels of Ha-ras transcripts decreased during calcium-induced differentiation in primary newborn epidermal cells. Differentiation was induced by switching the adult and newborn keratinocytes from medium containing 0.05 mM Ca2+ to medium containing one of three different calcium concentrations (0.15, 0.5, or 1.2 mM Ca2+). The decrease in Ha-ras mRNA levels observed during differentiation in newborn keratinocytes occurred as an intermediate event in the differentiation process, was specific for the Ha-ras gene, and was not due to a general decrease in transcriptional activity during differentiation. Characteristic patterns of keratin 14 gene expression and cornified envelope formation were observed, verifying that the differentiation process had been induced in both the primary adult and newborn epidermal cells. That adult keratinocytes are resistant to the differentiation-induced reduction in Ha-ras mRNA expression observed in newborn keratinocytes may explain the difference in in vivo tumorigenic potentials of newborn and adult skin.


Tissue-specific expression and induction of human O6-alkylguanine-DNA alkyltransferase in transgenic mice.

O6-Alkylguanine-DNA alkyltransferase (ATase) activity was determined in crude sonicates of tissues obtained from the F2 offspring of human ATase transgenic founder mice. In certain cases, samples were analyzed both before and after administration of zinc sulfate in the drinking water for 2 wk to upregulate the mouse metallothionein-1 promoter that controls the expression of the transgene. In liver samples obtained by partial hepatectomy, the ATase activities of nontransgenic mice ranged from 63 to 139 fmol/mg total protein (mean of 10 mice, 95.3 +/- 23 fmol/mg), whereas in positive transgenic mice, the range was from 503 to 2119 fmol/mg (mean of 10 mice, 963 +/- 475 fmol/mg). The difference between the mean ATase values for these two groups of mice is highly significant (P less than 0.001). All positive mice expressed ATase and in those examined using the human ATase coding sequence as a probe, isoschizomeric-restriction endonuclease digestion showed no evidence of cytosine methylation in the transgene. After zinc sulfate induction, the ATase levels in residual liver tissue were for the controls 84-191 fmol/mg (mean of 10 mice, 123 +/- 31.5 fmol/mg) and for positive mice 908-3273 fmol/mg (mean of 10 mice, 1960 +/- 724 fmol/mg). Induction thus caused a 1.4- to 3.2-fold increase in ATase activity in the tissues of individual transgenic mice (mean, 1.8-fold; P less than 0.003), with the greatest increase generally occurring in those mice that had the lowest preinduction levels. Hepatic ATase levels were thus increased up to 28 times higher in transgenic mice than in nontransgenic mice. When data from other groups of transgenic and nontransgenic mice (eight of each) was included and analyzed in an independent rather than paired fashion, the mean values for zinc-treated controls and transgenic mice, respectively, were 106 fmol/mg and 1415 fmol/mg, still a highly significant (P less than 0.001) difference. In two mice given a single intraperitoneal dose of cadmium chloride, hepatic ATase increased 2.1- and 4.9-fold, respectively. The effect of partial hepatectomy alone was also considered: for transgenic mice the mean ATase level increased from 453 to 661 fmol/mg protein after 48 h. In other offspring subjected to either unilateral nephrectomy or orchidectomy, induction of ATase activity by zinc sulfate was also seen in kidney (5.7- and 8.4-fold) and testis (1.7- and 3.1-fold), although these observations were made with small numbers of mice.(ABSTRACT TRUNCATED AT 400 WORDS)


Mutagenesis by apurinic sites in normal and ataxia telangiectasia human lymphoblastoid cells.

We used a shuttle vector based on the Epstein-Barr virus origin of plasmid replication (oriP) to determine the types of mutations induced by depurination in human cells. Plasmid DNA was incubated at pH 2 at 40 degrees C for various times to induce up to 20 apurinic (AP) sites per 9.7-kb plasmid and electroporated into lymphoblastoid cells derived from either a normal individual or an ataxia telangiectasia patient. After replication of the vector in the human cells, plasmid DNA was isolated and analyzed for mutations induced in the plasmid-encoded herpes simplex virus type 1-thymidine kinase gene. Both the frequencies and types of mutations induced by depurination were essentially identical for normal and ataxia telangiectasia cells. The mutant frequency at 20 AP sites/plasmid was 10-fold to 13-fold greater than that observed for untreated DNA. Deletion and frameshift events accounted for 46-55% of the mutants induced by depurination. The induced deletions were relatively small (median size, 100-150 bp) and characterized by short (1-5 bp) regions of sequence homology at the endpoints. These mutations and the frameshifts, a majority of which occurred in runs of identical nucleotides, are consistent with a model involving AP-site-induced template dislocation during DNA synthesis. A broad spectrum of base-substitution mutations, which accounted for 19-36% of the induced mutants, was observed. The apparent preference for insertion opposite AP sites in human cells was G (43-55%) greater than A approximately C (18-21%) greater than T (9-14%). Our results in human cells contrast markedly with those published previously for the mutational specificity of AP sites in Escherichia coli, in which a large majority of the mutants resulted from insertion of an A opposite the abasic site.


Analysis of point mutations in murine c-Ha-ras of skin tumors initiated with dibenz[a,j]anthracene and derivatives.

This study was designed to evaluate the point mutations in the murine c-Ha-ras gene of skin papillomas induced by initiation with dibenz[a,j]anthracene (DB[a,j]A), its bay-region anti-diol epoxide ((+/-)anti-DB[a,j]A-DE), and a 7,14-dimethyl analogue (7,14-diMeDB[a,j]A). Recent studies (Nair RV, et al., Chem Res Toxicol 4:115-122, 1991) in our laboratory have revealed both deoxyguanosine (dGuo) and deoxyadenosine (dAdo) adducts formed from the anti- and syn-diol epoxides of DB[a,j]A in cultured mouse epidermal cells after exposure to this hydrocarbon. Using PCR amplification and direct sequencing, we found specific A182----T transversion mutations (eight of 10 tumors) in codon 61 of c-Ha-ras in papillomas induced by initiation with DB[a,j]A. Analysis of papillomas generated by initiation with the more biologically potent analogue 7,14-diMeDB[a,j]A revealed that five of five tumors exhibited A182----T transversions in codon 61. The nature of the changes in the two DB[a,j]A tumors not showing codon 61 mutations in Ha-ras is currently not known since these tumor DNAs also did not possess c-Ha-ras mutations at codons 12, 13, or 59. Interestingly, papillomas produced by initiation with (+/-)anti-DB[a,j]A-DE also possessed A182----T transversion mutations in codon 61 of c-Ha-ras (five of five tumors). These data suggest that dAdo adducts derived from both parent hydrocarbons may play an important role in their tumor-initiating activity and possibly implicate a specific diol epoxide-dAdo adduct in this process.


Activation of the Ki-ras gene in spontaneous and chemically induced lung tumors in CD-1 mice.

As part of an evaluation of the effectiveness of using ras mutation analysis for distinguishing carcinogen-induced from spontaneous tumors, we examined the profile of ras gene point mutations in spontaneous, 7,12-dimethylbenz[a]anthracene (DMBA)-induced, and N-nitrosodiethylamine (DEN)-induced lung tumors from Crl:CD-1(ICR)BR (CD-1) mice. Although all of the lung tumors were assayed for mutations in the Ha-ras, Ki-ras, and N-ras genes (codons 12, 13, and 61), only Ki-ras mutations were found, which is consistent with other studies that have noted a strong preference for Ki-ras gene activation in mouse, rat, and human lung tumors. We found that spontaneous CD-1 mouse lung tumors had a very high frequency of Ki-ras gene activation (17 of 20 tumors; 85%), distributed among codons 12 (5 of 20), 13 (1 of 20), and 61 (11 of 20). DMBA-induced lung tumors had a slightly higher frequency of Ki-ras gene mutations (16 of 16; 100%), again distributed among codons 12 (5 of 16), 13 (2 of 16), and 61 (9 of 16). However, seven of the DMBA tumors had mutations qualitatively different from those found in spontaneous tumors. In contrast to DMBA-induced tumors, DEN-induced tumors had a lower frequency of Ki-ras mutations (36%) when compared with spontaneous lung tumors, suggesting that DEN primarily induces lung carcinogenesis by a mechanism other than ras gene activation. Thus, although spontaneous and induced CD-1 mouse lung tumors have a strong tissue-specific preference for carrying an activated Ki-ras gene, the nature of the initiating carcinogen can influence the frequency or profile of Ki-ras mutations.


Fetal abdominal hyperechoic mass: diagnosis and management.

In the last 2 years, we have had the opportunity to follow 12 cases diagnosed with hyperechoic abdominal masses. Four of the cases ended with a fetal demise, while 7 resulted in the birth of an anomalous or medically ill neonate. Only 1 case has shown spontaneous resolution of the hyperechoic mass with the birth of a normal neonate. The in utero diagnosis of a hyperechoic abdominal mass should encourage the clinician to further investigation since the differential diagnosis is quite diverse. Appropriate counselling for the patient is a necessary part of prenatal care.


Fetal nuchal oedema: associated malformations and chromosomal defects.

During an 8-year period, oedema in the dorsal cervical region that produces a characteristic tremor on ballotement of the fetal head (nuchal oedema) was observed in 145 (7%) of the 2,086 fetuses that underwent karyotyping in our unit because of fetal malformations and/or growth retardation. Nuchal oedema was distinguished from nuchal cystic hygromata and from hydrops foetalis. In 132 (91%) of the cases with nuchal oedema, there were other fetal malformations, and 53 (37%) fetuses had chromosomal abnormalities, mainly trisomy 21 but also other trisomies, deletions or translocations, triploidy and Turner syndrome. Furthermore, the chromosomally normal fetuses with nuchal oedema had a very poor prognosis because, in many cases, there was an underlying skeletal dysplasia, a genetic syndrome or cardiac defect.


Strawberry-shaped skull in fetal trisomy 18.

During an 8-year period, a strawberry-shaped skull (flattening of the occiput with pointing of the frontal bones), was observed in 54 (3%) of the 2,086 fetuses that underwent karyotyping in our unit because of fetal malformations and/or growth retardation. In all 54 cases with a strawberry-shaped skull, there were other fetal malformations; in 43 (80%) fetuses, there was trisomy 18 and in 1 triploidy. Therefore, the ultrasonographic finding of a strawberry-shaped skull should initiate a diligent search for the presence of other markers of trisomy 18 and is a strong indication for fetal karyotyping. However, in the total series of 2,086 fetuses with malformations and/or growth retardation, there were another 40 fetuses with trisomy 18 and 41 with triploidy who did not have a strawberry-shaped skull.


Fetal cardiovascular physiology and therapy.

The ability to diagnose fetal cardiovascular malformations prenatally has been established during the past 14 years. This has naturally led to an increased interest in the possibility of providing timely therapy in the neonatal period and has raised the prospects for prenatal intervention. Before medical, catheter, and/or surgical interventions are performed, however, it is important to understand the normal and abnormal physiology of the fetal cardiovascular system. Significant insights have been gained into human fetal cardiovascular physiology utilizing data previously gleaned from fetal lamb models and correlating anatomic, pulsed Doppler, and color flow Doppler observations that have been made echocardiographically during the second and third trimesters of human fetal cardiac development. Regional blood flow distribution studies in the human have demonstrated a relative right-ventricular volume dominance in the human fetus of a somewhat lesser magnitude than previously described in the lamb. Observations of ventricular filling characteristics suggest a relative paucity of diastolic 'reserve' in the human fetal heart implying a relative sensitivity to acute volume overloading. The human fetal heart, like that of the fetal lamb, also appears to have a relatively modest amount of 'systolic reserve', making the heart particularly susceptible to acute ventricular afterload. Further studies have demonstrated the ability to recognize altered intrauterine shunt flow across the fetal ductus arteriosus and foramen ovale. As predicted in fetal lamb models, alterations in shunt flow accompany major malformation complexes including ventricular hypoplasia. Studies to date suggest that attempts at surgical intervention should be made very cautiously due to the inability to ascertain whether altered shunt flow patterns precede (cause?) or follow the development of structural abnormalities.


Early prenatal diagnosis and therapy of fetal hypothyroid goiter.

We report a case of early diagnosis of iodide-induced fetal hypothyroidism at 22 weeks of gestation, confirmed at 29 weeks by cordocentesis and successfully treated intra-amniotically. The ultrasonographic feature was the presence of two echogenic masses in the fetal neck; polyhydramnios was absent. Mild hypothyroidism was diagnosed based on fetal serum obtained by percutaneous umbilical blood sampling at 29 weeks of gestation. The persistence of fetal hypothyroidism in spite of maternal thyroid improvement was confirmed by a second cordocentesis at 35 weeks of gestation, and a single injection of intra-amniotic levothyroxine (250 micrograms) was performed. The serial ultrasonographic examinations showed disappearance of the fetal goiter. A healthy female baby (3,630 g) was delivered at term. At birth, the thyroid gland was not enlarged, and neonatal thyroid hormones were within the normal range. This case suggests that cordocentesis is a reliable method to assess the fetal thyroid status; moreover a single injection of intra-amniotic thyroxine was effective in treating fetal hypothyroid goiter.


Fetal therapy, ethics and public policies.

This article reviews the evolution of ethical problems in the first generation of experimental fetal therapy and the prevailing approaches to them. The problems include: review of risks/benefits, case selection, informed consent, twin pregnancies, and refusal of proven fetal therapy. The article further discusses ethical and public policy issues in the lack of U.S. federal support for fetal diagnosis, fetal therapy, and human embryo research. An argument is made for such support, beginning with experimental gene therapy in the fetus. Ethical principles are identified that support an obligation to learn to relieve and treat such human suffering at the earliest time. Contradictions of these principles are also identified in terms of research not now supported.


Intrathoracic silo for the potential antenatal repair of diaphragmatic herniae with liver in the chest.

Antenatal correction of congenital diaphragmatic hernia in the human fetus is an attractive theoretical possibility that might reverse pulmonary hypoplasia before it becomes too severe for post-natal survival. Initial attempts at correcting these herniae in the human fetus have been beset with many technical problems, one of which has been to prevent acute obstruction to the ductus venosus when the left lobe of the liver is reduced back into the abdomen. We describe a new procedure intended to reduce the liver gradually so that the ductus venosus is never acutely kinked and more normal rib cage mechanics may be maintained.


First trimester interfetal size variation in well-dated multifetal pregnancies.

Sonographic measurements of multifetal pregnancy for dating and growth are considered identical to singleton pregnancies until the second trimester. Observations in 57 patients having triplets or more, who were referred for first trimester multifetal pregnancy reduction, suggest considerable size variability both within an individual pregnancy and among pregnancies of the same gestational ages (GAs). These data are unique because the GAs are precisely known, because the pregnancies were established by assisted reproductive technology. We observed a greater standard deviation at 12 and 13 weeks of GA. We also observed significant interfetal variation within an individual pregnancy. Because multifetal pregnancy reduction was performed, follow-up of any given fetus to term was not possible. We conclude that for high-order multiple gestations: (1) interfetal size variability begins much earlier than often stated; (2) differences in the degree of variability can be observed among different pregnancies of the same gestational age and within the same pregnancy, and (3) there is a trend for increasing variability with increasing gestational age.


Fetal cerebral blood flow velocity during labour. Preliminary report.

The authors have tried to ascertain in a preliminary study if monitoring of fetal cerebral blood flow velocity during labour could identify a situation threatening the fetal brain, which could justify a compulsory fetal extraction. Nine patients were explored at term during labour. The Doppler signal was recorded using the abdominal route at the level of the fetal internal carotid. The diastolic carotid index was used (DCI = D/S). In 5 cases, fetal heart rate (FHR) was normal during labour and the DCI was on average comparable for each of the patients and the mean DCI (20.1 +/- 1%) was in close agreement with the values normally seen after 33 weeks of amenorrhoea during pregnancy. The case of isolated deceleration shows that deceleration causes an almost instant drop in the telediastolic flux, but the index increases more rapidly than could be expected from the increase in cardiac frequency if this phenomenon were of purely cardiogenic origin. In the case of bradycardia, it was noticed that during the return to the initial FHR frequency, the DCI was much higher than during the measurements performed before bradycardia. In the 1st case of pathological FHR, the index was very high and remained elevated during all the observation period. The fetus presented at birth a normal Apgar score as well as normal blood pH and neurological examination. In the 2nd case of pathological FHR, the FHR was abnormal with decelerations, the DCI increased rapidly and remained high.(ABSTRACT TRUNCATED AT 250 WORDS)


Ventrolateral prefrontal cortex lesions in rats impair the acquisition and retention of a tactile-olfactory configural task.

Rats with aspirative lesions of the ventrolateral frontal cortex were tested on acquisition and postsurgical retention of an associative learning task that required that they learn a tactile-olfactory configural discrimination. The task required that they pull up a string to obtain attached food and that they identify the correct string using a compound of string size and odor. The rats were not impaired in initial learning or reversal of the olfactory elements of the discrimination. They were impaired in acquisition and retention of the compound, and their deficit was proportional to lesion size. The results confirm that the ventrolateral frontal cortex is involved in processing of olfactory information and imply that the prefrontal cortex is involved in at least certain types of cross-modal configural associative learning.


Visual and auditory spatial and nonspatial delayed-response performance by Korsakoff and non-Korsakoff alcoholic and aging individuals.

Thirty-six male alcoholics (13 with Korsakoff's syndrome) and 24 controls performed visual and auditory delayed-response tasks sensitive to prefrontal cortical damage in nonhuman primates. Korsakoff patients were consistently impaired compared with other subjects. Impairments by Korsakoff patients were evident when demands were placed on visual processing time (brief stimulus durations), and the deficits became exaggerated with increased demands on short-term memory. Under the most difficult experimental conditions, controls and non-Korsakoff alcoholics who were over 50 years old performed somewhat worse compared with younger groups 27-49 years old. Age-linked deficits were mild compared with Korsakoffs' deficits, and age-group differences disappeared with easier task demands. The results implicate cortical pathology in alcoholism and normal chronological aging and suggest that prefrontal damage accompanies alcoholic Korsakoff's syndrome.


Radio-frequency lesions of the thalamus produce delayed-nonmatching-to-sample impairments comparable to pyrithiamine-induced encephalopathy in rats.

Rats were trained and matched on a delayed-nonmatching-to-sample (DNMTS) task and randomly assigned to treatment. In Experiment 1, radio-frequency (RF) lesions were aimed at lateral portions of the internal medullary lamina (L-IML), midline thalamus (MT), mammillary bodies (MB), and the combination of MT and MB. In Experiment 2, RF lesions were aimed at the fornix. After recovery, DNMTS was retrained at retention intervals retention interval of 3.0-18.0 s, the critical retention interval for 75% DNMTS accuracy was determined by a staircase procedure, and spontaneous exploration was observed in an open field. L-IML lesions produced significant deficits on DNMTS and exploratory behavior that were comparable to deficits on the same tasks in rats recovered from pyrithiamine-induced thiamine deficiency. Fornix lesions produced significant DNMTS deficits that were substantially smaller than for the L-IML group. The MT, MB, and MT+MB treatments had no significant effect on DNMTS.


Eyeblink conditioning in the developing rat.

Long-Evans rat pups, 17-18 or 24 days of age, were trained with an eyeblink conditioning (EBC) procedure that was used previously with adult rats (Skelton, 1988). Pups received 3 sessions of delay conditioning in a single day at about 4-hr intervals (100 trials/session). Trials involved pairings of an auditory conditioned stimulus (2.8-kHz, 82-dB tone) and a periocular-shock unconditioned stimulus (US; 100 ms, 2 mA), which were presented 280 ms apart. EBC was observed at both ages, but older pups learned much more rapidly. Subsequent experiments established that this effect is associative (Experiment 2), that age differences in EBC cannot be attributed to differences in ability to respond or in sensitivity to the US (Experiment 3), and that EBC rate can be modulated by motivational state (Experiment 4). This preparation may help elucidate the relation between neural development and the ontogeny of learning.


Learning-dependent timing of Pavlovian eyelid responses: differential conditioning using multiple interstimulus intervals.

This study demonstrates that individual animals can concurrently acquire differently timed conditioned eyelid responses using a differential conditioning procedure in which distinctive conditioned stimuli (CSs) are individually paired with an unconditioned stimulus, with each using a different interstimulus interval (ISI). This promotes robust conditioning, and the timing of the conditioned responses is appropriate for the respective ISIs, differs for each CS to the extent that the ISIs are dissimilar, and is apparent in individual trials. This procedure was used to demonstrate that response timing is not a function of associative strength. These data suggest response timing is mediated by an ability to make temporal discriminations during the CS. The within-animals comparisons made possible by this differential conditioning should facilitate lesion and unit recording analyses of the neural basis of response timing.


Pelvic and pudendal nerves influence the display of paced mating behavior in response to estrogen and progesterone in the female rat.

Two experiments examined whether the pudendal and pelvic autonomic nerves are important for pacing behavior shown by the female rat during mating. Ovariectomized female rats received bilateral transection of the pudendal (Pux), pelvic (Pex), or pudendal and pelvic (Pu + Pex) nerves or were sham-operated (Shx). Lordotic behavior, precopulatory solicitations, postural adjustments, and pacing behavior were measured 14 days after nerve transection in Experiment 1 after sequential treatment with estradiol benzoate (EB) and progesterone (P) and in Experiment 2 on the day after 7 daily injections of EB. After combined EB and P treatment, disruption of pacing behavior occurred in Pex and Pu + Pex animals. After EB-only treatment, Pux animals as well as Pex and Pu + Pex animals showed decreased pacing behavior. Thus, afferent input via the pelvic nerve is important for the display of pacing behavior, and P may counteract the effects of autonomic nerve transection.


Effects of unilateral hypothalamic manipulations on the sexual behaviors of rats.

Four experiments that assessed the contributions of each side of the hypothalamus to the control of sexual behavior found the following. (a) Exposing the left, but not the right, ventromedial nucleus to estrogen neonatally defeminized sexual behavior in female rats. This asymmetry did not reverse as sexual differentiation progressed. (b) Unilateral cuts lateral to the medial preoptic area disrupted mounting in females that had mounted regularly before surgery, when given testosterone. The deficits were greater when the cuts were on the left side, but a third of the females with unilateral cuts showed severe deficits regardless of the side. (c) Comparable cuts did not impair masculine sexual behavior in gonadally intact males. In fact, left-side cuts seemed to accelerate copulation in males. (d) Unilateral lesions of the ventromedial nucleus disrupted lordosis in female rats in an essentially all-or-none fashion. This effect did not vary with side.


Control of ingestion by oral habituation in rat pups.

The role of habituation of mouthing activity in the control of ingestion was investigated in 6-, 12-, and 18-day-old rat pups. In pups at all ages, oral habituation to a flavored diet inhibited ingestion of a continuous oral infusion of that same diet. Twelve-day-old pups that had orally habituated to a diet continued to consume less of a continuous oral infusion of that diet both 30 min and 3 hr later, and the duration of suppressed ingestion was shown to be dependent on the rate of stimulus presentation during habituation experience. These data suggest that oral habituation may be a diet-specific influence on both intra- and intermeal patterning.


Depletion of cortical norepinephrine in rats by 6-hydroxydopamine does not impair performance of a delayed-nonmatching-to-sample task.

Rats were trained on a spatial delayed-nonmatching-to-sample (DNMTS) task, matched for performance, and randomly assigned to treatment with dorsal noradrenergic bundle injections of either 6-hydroxydopamine, to deplete cortical norepinephrine (NE), or vehicle, to control for the effects of surgery. After recovery, there were no significant differences between the groups when retrained on the DNMTS task at retention intervals (RI) from 0.1 to 15.0 s. Furthermore, no differences were observed when rats were trained at a 6.0-s RI filled with distracting stimuli or when dummy information runs were added to increase proactive interference. These results demonstrate that depletion of cortical NE cannot account for the DNMTS performance deficits observed in rats recovered from pyrithiamine-induced thiamine deficiency (Knoth & Mair, 1991; Robinson & Mair, 1992).


HLA-associations in Leponex/Clozaril (clozapine)-induced granulocytopenia and agranulocytosis. Statistical viewpoints.

This paper describes a statistical analysis of the simultaneous occurrence of 48 human leucocyte antigens (HLAs) in an attempt to predict whether a patient treated with clozapine will develop granulocytopenia or agranulocytosis. Whereas the numerical results show that use of HLAs does not appear to satisfy the intended purpose, emphasis is placed on the statistical methodology recommended for a problem of this type. Other work in this area is critically discussed.