Kinesin and myosin ATPases: variations on a theme.

The enzymes kinesin and myosin are examples of molecular motors which couple ATP hydrolysis to directed movement of biological structures. Myosin has been extensively studied and its structure and mechanism of coupling are known in detail. Much less is known about kinesin, but many of its major properties are similar to those of myosin. Both enzymes have two catalytic head groups at the end of a long alpha-helical rod. The head groups contain the sites for ATP hydrolysis and interaction with their respective partners for movement (microtubules or F-actin). In each case the binding and hydrolysis of ATP is rapid and the steady state ATPase rate is limited by a slow step in the region of product release. This slow release of product is accelerated by interaction with actin or microtubules coupled to changes in binding affinity. As there is no evidence for a close evolutionary link between kinesin and myosin, these and other similarities may represent convergence to set of common functional properties which are constrained by the requirements of protein structure and the use of ATP hydrolysis as a source of energy. It will be of particular interest to determine if these common properties are also shared by the large number of divergent proteins which have recently been discovered to possess a domain which is homologous to the head group of kinesin.


The role of nucleoside triphosphate hydrolysis in transducing systems: p21ras and muscle.

A variety of systems use nucleoside triphosphate hydrolysis to control or provide energy for biological processes, mediated through protein-protein interactions. The nature of this coupling may vary, but often there is a degree of similarity. In this paper, two systems are compared: actomyosin in muscle and p21ras in a signal transduction pathway as yet undefined. The mechanism of the nucleotide triphosphate hydrolysis and the consequent changes in the protein-nucleotide complex have been investigated, to understand how the coupling to biological function is achieved. The basal nucleoside triphosphatase mechanisms are compared and the roles of proteins that activate the hydrolysis, actin and GAP, are discussed. The cleavage process was probed by stereochemical techniques to determine the basic mechanism, of either a phosphorylated enzyme intermediate or direct displacement of nucleoside diphosphate by water. Phosphate-water oxygen exchange probes were used to investigate nucleoside triphosphate and inorganic phosphate release steps. A new method of probing the kinetics of inorganic phosphate release directly has been developed. In muscle, this process seems likely to be related directly to force generation. In the GAP-ras system, measurement of phosphate release is allowing the mechanism of the GAP-p21ras interaction to be probed.


Nucleotides and divalent cations as effectors and modulators of exocytosis in permeabilized rat mast cells.

The idea that the universal trigger to exocytosis (the terminal step in the secretory process) is an elevation of the cytosol concentration of Ca2+, and that it is dependent on ATP, is no longer tenable. Working with streptolysin-O-permeabilized mast cells (and other myeloid cells) we have shown that non-hydrolysable analogues of GTP can stimulate exocytosis after depletion of Ca2+ (i.e. at concentrations below 10(-9) M) and ATP. Such Ca2+- and ATP-independent exocytosis is strongly dependent on the presence of Mg2+, and the requirement for Mg2+ declines as the concentration of Ca2+ is brought up to 10(-7) M. We argue that Ca2+ serves to regulate the binding of guanine nucleotides to GE, a GTP-binding protein that regulates exocytosis through its interaction with CE, a calcium-binding protein which serves as an intracellular pseudo-receptor. The onset of exocytosis, following provision of Ca2+ and guanine nucleotides to the permeabilized cells, is preceded by delays which are sensitive to the order of provision of the two effectors (i.e. Ca2+ and guanine nucleotides), the presence or absence of Mg2+, and the identity of the activating guanine nucleotide. In view of the similarity of these features with the activation kinetics of adenylyl cyclase, we argue that GE behaves as a member of the heterotrimeric class of signal transducing G-proteins such as GS.


Studies on the structure and mechanism of H-ras p21.

Current knowledge of the structure of H-ras p21 is reviewed with particular emphasis on the interaction between guanine nucleotides and the active site of the protein. The nature of the conformational change induced by GTP hydrolysis is discussed. The major change is seen in the region known as the effector loop (loop 2), with significant but less well-defined changes occurring in loop 4, which is implicated in the GTPase reaction. Other evidence concerning the mechanism of GTP hydrolysis and its activation by GAP (GTPase-activating protein) is also discussed. Evidence regarding the rate limiting step in the p21 GTPase reaction, and the manner in which this and possibly other steps are accelerated by GAP, is inconclusive.


Hormone signalling via G-protein: regulation of phosphatidylinositol 4,5-bisphosphate hydrolysis by Gq.

Heterotrimeric GTP-dependent regulatory proteins (G-proteins) mediate modulation by many cell surface receptors. Activation of the G-proteins promotes dissociation of their alpha and beta gamma subunits. The similarity of behaviour of beta gamma subunits derived from a variety of G-proteins has led to their use as affinity reagents for the analysis of the more unique alpha subunits. The evolution of these uses is presented. One of the more insightful results was the isolation of a new class of G-protein alpha subunits (the alpha q subfamily) which have been shown to regulate the activity of a phospholipase C (PLC) specific for phosphatidylinositols. The experimental evidence leading to this conclusion is discussed. The activation by alpha q increases the apparent Vmax of the beta isoform of phosphatidylinositol-specific phospholipase C (PLC beta) and can be modulated by beta gamma subunits.


Coupling of ras p21 signalling and GTP hydrolysis by GTPase activating proteins.

Ras p21 proteins cycle between inactive, GDP-bound forms and active GTP-bound forms. Hydrolysis of bound GTP to GDP is mediated by proteins referred to as GAPs, two forms of which have been described. The first, p120-GAP, contains regions of homologies with tyrosine kinase oncogenes, and interacts with tyrosine phosphoproteins as well as with ras proteins; p120-GAP may therefore connect signalling pathways that involve tyrosine kinase and ras p21 proteins. The second type of GAP is the product of the neurofibromatosis type 1 gene (NF1-GAP). This is a protein of 325,000 Da that is defective in patients with NF1; NF1-GAP is regulated by signalling lipids, and may serve to connect ras p21 with phospholipid second messenger systems. The significance of ras p21 interaction with distinct GAPs is discussed.


The kinetic mechanism of the GAP-activated GTPase of p21 ras.

Guanine nucleotides modified by acetylation of the ribose moiety with the small fluorophore N-methylanthranilic acid(mant) have been shown to bind to p21 ras with similar equilibrium and kinetic rate constants as the parent nucleotides. Hydrolysis of p21.mantGTP to p21.mantGDP results in a 10% decrease in fluorescence intensity occurring at the same rate as the cleavage step. A similar process occurs with the non-hydrolysable analogue mantGMP.PNP, and this has led to the proposal that a conformational change of p21.mantGTP precedes and controls the rate of the cleavage step. The fluorescence change with p21.mantGMP.PNP is accelerated in the presence of the C-terminal catalytic domain of GAP, which is consistent with this mechanism. The same conformational change does not occur with oncogenic mutants of p21 ras, Asp-12 and Val-12, but does occur with the weakly oncogenic Pro-12 mutant. Stopped flow measurements of the interaction of GAP with p21.mantGTP show an exponential decrease in fluorescence, the rate of which does not vary linearly with GAP concentration. These data imply a rapidly reversible formation of the p21.mantGTP complex with GAP followed by the isomerization of this complex. This is at least 10(5)-fold faster than the same process in the absence of GAP.


Photochemical mapping of the active site of myosin.

The active sites of myosin from skeletal, smooth and scallop muscle have been partly characterized by use of a series of photoreactive analogues of ATP. Specific labelling was attained by trapping these analogues in their diphosphate forms at the active sites by either cross-linking two reactive thiols (skeletal myosin) or by formation of stable vanadate-metal ion transition state-like complexes (smooth muscle and scallop myosin). By use of this approach combined with appropriate chemistry, several key residues in all three myosins have been identified which bind at or near the adenine ring, the ribose ring and to the gamma-phosphate of ATP. This information should aid in the solution of the crystal structure of the heads of myosin and in defining a detailed structure of the ATP binding site.


The actomyosin ATPase: a two-state system.

Studies of the interaction between actin and myosin subfragment 1 (S1) in solution have shown that the association reaction takes place in at least two steps. Initially the association is relatively weak to form a complex called the A state which can then isomerize to the R state. The rate and equilibrium constants for the isomerization have been measured and are shown to depend upon the nucleotide bound to the S1 ATPase site; with ATP bound the A state is preferred but as ATP is hydrolysed and the products are sequentially released then the complex gradually shifts to the A state. An extensive series of experiments have characterized the A-to-R isomerization both in solution and in contracting muscle fibres and have shown it to be closely associated with the key events in the ATP-driven contraction cycle: the conformational change from the A to the R state can be monitored by fluorescent probes on either actin or the nucleotide; the isomerization can be perturbed by increases in hydrostatic pressure; the actin-induced acceleration of the rate of product release from myosin is coupled to the A-to-R isomerization; tropomyosin may control actin and myosin interaction by controlling the isomerization step and finally pressure perturbations of contracting muscle fibres shows there to be a close coupling between the isomerization of acto.S1 and the force generating event of muscle contraction.


Nitrogenase of Klebsiella pneumoniae: an MgATP hydrolysing energy transduction system with similarities to actomyosin and p21 ras.

The mechanism of ATP hydrolysis by nitrogenase shows some similarity to that proposed for actomyosin and for GTP hydrolysis by p21 ras. All three systems involve the formation of an active complex from two component proteins, nucleotide-induced changes in protein conformation, energy transduction that in the case of nitrogenase involves a decrease in redox potential of metal centres, and a slow dissociation of the protein complex. Metal ion activation (Mg2+ or Ca2+) and in-line displacement of ADP by H2O without enzyme phosphorylation are also common features. At 5 degrees C, stopped-flow calorimetry shows that the kinetic and thermodynamic parameters for endothermic, reversible on-enzyme cleavage of MgATP by nitrogenase and myosin subfragment 1 are remarkably similar. [18O4]Pi-water exchange studies also show that ATP cleavage on nitrogenase and myosin are reversible.


DNA supercoiling and relaxation by ATP-dependent DNA topoisomerases.

Bacterial DNA gyrase and the eukaryotic type II DNA topoisomerases are ATPases that catalyse the introduction or removal of DNA supercoils and the formation and resolution of DNA knots and catenanes. Gyrase is unique in using ATP to drive the energetically unfavourable negative supercoiling of DNA, an example of mechanochemical coupling: in contrast, eukaryotic topoisomerase II relaxes DNA in an ATP-requiring reaction. In each case, the enzyme-DNA complex acts as a 'gate' mediating the passage of a DNA segment through a transient enzyme-bridged double-strand DNA break. We are using a variety of genetic and enzymic approaches to probe the nature of these complexes and their mechanism of action. Recent studies will be described focusing on the role of DNA wrapping on the A2B2 gyrase complex, subunit activities uncovered by using ATP analogues and the coumarin and quinolone inhibitors, and the identification and functions of discrete subunit domains. Homology between gyrase subunits and the A2 homodimer of eukaryotic topo II suggests functional conservation between these proteins. The role of ATP hydrolysis by these topoisomerases will be discussed in regard to other energy coupling systems.


Nucleotide hydrolysis regulates the dynamics of actin filaments and microtubules.

Actin filaments and microtubules are major dynamic components of the cytoskeleton of eukaryotic cells. Assembly of these polymers from monomeric actin or tubulin occurs with expenditure of energy, because ATP (or GTP) tightly bound to actin (or tubulin) is irreversibly hydrolysed during polymerization. Therefore, actin filaments an microtubules are dissipative structures. Our purpose has been to understand how the dissipation of chemical energy perturbs the laws of reversible helical polymerization defined by Oosawa, and affects the dynamics of these polymers. A kinetic study has shown that nucleotide is hydrolysed on the polymer within at least two steps consecutive to the incorporation of the monomer: cleavage of the gamma-phosphoester bond followed by the slower release of Pi; only the second reaction appears reversible. Pi release, and not cleavage of the gamma-phosphate, is linked to the destabilization of protein-protein interactions in the polymer, and therefore plays the role of a conformational switch. The dynamic properties of the polymer in the NTP- and NDP-Pi intermediate states of the assembly process have been investigated using non-hydrolysable analogues of nucleotides and structural analogues of Pi, AlF4- and (BeF3-, H2O). Because nucleotide hydrolysis is uncoupled from polymerization, actin filaments and microtubules grow with a 'cap' of terminal NTP- and NDP-Pi-subunits that interact strongly, and prevent the rapid depolymerization of the unstable core of the polymer formed of NDP-subunits. The fact that the dynamic properties of the polymer are affected by bound nucleotide results in a nonlinear dependence of the rate of elongation on monomer concentration.(ABSTRACT TRUNCATED AT 250 WORDS)


Self-organization of polymer-motor systems in the cytoskeleton.

Microtubules and actin filaments are organized into dynamic arrays inside cells. In this paper I discuss in conceptual form the assembly mechanisms of three specific arrays: asters, spindles and leading edge structures. The role of energy transducing processes, particularly motor protein activity, in assembly is explored. I conclude that dynamic interaction between motor proteins and cytoskeletal polymers play a very general role in spatial organization of the cytoplasm.


Flesinoxan shows antidepressant activity in a DRL 72-s screen.

Schedules which selectively reinforce low rates of responding (DRL, differential reinforcement of low rate) distinguish between antidepressants and other types of drugs. In a DRL schedule a subject is required to pause for a specified minimum period of time between two consecutive responses in order to obtain a reinforcer. The dependent variables are rate of responding and rate of reinforcement. Response patterns of rats treated with clinically effective antidepressant drugs such as imipramine (2.0-32.0 mg/kg) or fluvoxamine (4.0-32.0 mg/kg) are characterized by a decrease in response rate and an increase in reinforcement rate. Treatment with the 5-HT1A agonist flesinoxan (0.1-3.0 mg/kg) also dose-dependently decreased response rates while at the same time increasing reinforcement rates. Chlordiazepoxide (2.5-20.0 mg/kg) and diazepam (0.25-2.0 mg/kg) had no effects in the present experiment. d-Amphetamine increased response rates at low doses (0.5-2.0 mg/kg), and decreased it at the higher doses (4.0 mg/kg), but reinforcement rates were unaltered. Overall analysis of the effects of haloperidol (0.02-0.32 mg/kg) showed decreased responding and increased reinforcement rates. Post hoc analysis, however, clearly differentiated between haloperidol's profile and that of the antidepressants. As such, the results of the present experiment show that flesinoxan might possess antidepressant activity in humans.


A comparison of various antidepressant drugs demonstrates rapid desensitisation of alpha 2-adrenoceptors exclusively by sibutramine hydrochloride.

The functional status of presynaptic and postsynaptic alpha 2-adrenoceptors in murine brain was respectively monitored using the hypoactivity (sedation) and mydriasis (pupil dilatation) responses to clonidine (0.1 mg/kg IP). Both responses were attenuated 24 h after 3 days of injection of sibutramine hydrochloride (3 mg/kg IP). To ascertain whether this property was exclusive to sibutramine, the following antidepressant drugs were also tested for their ability to down-regulate alpha 2-adrenoceptors rapidly: amitriptyline, doxepin, nomifensine, desipramine, amoxapine, fluoxetine, zimeldine, tranylcypromine and mianserin. When given for 3 or 5 days at the low dose of 3 mg/kg IP, none of the other antidepressants reduced clonidine-induced hypoactivity or mydriasis. Furthermore, increasing the dose of amitriptyline, doxepin, nomifensine, desipramine, amoxapine and tranylcypromine to 10 mg/kg IP did not enable these antidepressants to attenuate the alpha 2-adrenoceptor-mediated responses after 3 days of treatment. An electroconvulsive shock (ECS; 200 V, 2 s) given once daily attenuated clonidine-induced mydriasis, but not hypoactivity, when administered for 3 days and both responses when administered for 5 days. In conclusion, this comparative study using antidepressant treatments with differing pharmacological modes of action demonstrated that sibutramine was the only drug which rapidly down-regulated pre- and postsynaptic alpha 2-adrenoceptors. ECS down-regulated postsynaptic alpha 2-adrenoceptors when given for 3 days, but required 5 days to desensitise both alpha 2-adrenoceptor populations.


Measurement of depression and negative symptoms in schizophrenia.

The validity of the Hamilton Depression Scale (HAM-D) as a measure of depressive symptomatology in schizophrenic patients is questionable since it was not developed for this purpose, nor has it been validated in a schizophrenic population. Accordingly, 80 schizophrenic inpatients were administered the HAM-D, the 18-item Brief Psychiatric Rating Scale (BPRS), and the Scale for the Assessment of Negative Symptoms (SANS) at drug-free baseline and after 4 weeks of neuroleptic treatment. The findings revealed that the HAM-D total score was nonspecific, while individual HAM-D subfactors provided a better index of various symptom complexes. The HAM-D contained a depressive factor that correlated strongly with the BPRS depression factor and a negative symptom factor that correlated strongly with the SANS and the BPRS negative symptom factor. These findings suggest the need to utilize specific assessment techniques rather than global measures when assessing depression in schizophrenia.


Estimation of time since exposure for a prevalent cohort.

In natural history studies of human immunodeficiency virus type 1 (HIV-1) infection a substantial proportion of participants are seropositive at time of enrollment in the study. These participants form a prevalent subcohort. Estimation of the unknown times since exposure to HIV-1 in the prevalent subcohort is of primary importance for estimation of the incubation time of AIDS. The subset of the cohort that tested negative for antibody to HIV-1 at study entry and was observed to seroconvert forms the incident subcohort that provides longitudinal data on markers of maturity (that is, duration) of infection. We use parametric life table regression models incorporating truncation to describe the conditional distribution (imputing model) of the times since seroconversion given a vector of the markers of maturity. Using the fitted model and the values of the markers of maturity of infection provided by the seroprevalent subcohort at entry into the study, we can impute the unknown times since seroconversion for the prevalent subcohort. We implement multiple imputation based on a model-robust estimate of the covariance matrix of parameters of the imputing model to provide confidence intervals for the geometric mean of the time since seroconversion in the prevalent subcohort, and to compare maturity of infection of cohorts recruited in different cities. The accuracy of imputation is further validated by comparisons of imputation-based estimates of AIDS incubation distribution in the seroprevalent subcohort with more direct estimates obtained from the seroincident subcohort.


[New positive inotropic drugs in acute and chronic heart failure].

Beta-adrenergic stimulants (Dobutamine and Dopamine) and recently introduced phosphodiesterase inhibitors (PDI) such as Amrinone, Milrinone, Enoximone and Piroximone are the principal inotropic agents for the treatment of acute cardiac failure. Most of the hemodynamic effects of these drugs are comparable, but peripheral vasodilatation is more marked with PDI. A potential advantage of the latter group is the lack of development tolerance, which occurs within 48 to 72 hours after beta-stimulants. On simultaneous administration, additive effects can be observed. Short term clinical results with PDI are good, especially in patients with postoperative cardiocirculatory failure, including cardiogenic shock. In contrast, long-term oral treatment with Amrinone, Milrinone and Enoximone in recent studies was disappointing. Efficacy was not superior to Digoxin, and unwanted side effects were frequent. Intermittent instead of continuous administration of positive inotropic agents should be evaluated in patients with severe congestive heart failure not responding to vasodilators and diuretics.


[Primary prevention of varicose bleeding: current status].

1. Surgical shunts should be abandoned for prevention of the first bleeding because of an increased mortality rate, despite a marked reduction in frequency of bleeding. 2. Sclerotherapy is not recommended for clinical practice in preventing the first bleeding. The meta-analysis of numerous randomized clinical trials shows a small effect on bleeding risk and mortality rate. But these effects are remarkably heterogenous in various studies and are compensated by the complications of sclerotherapy. 3. Prophylactic treatment with beta-blockers has similar results as sclerotherapy documented by meta-analysis of several studies. In contrast to sclerotherapy both risk and side effects are minimal. More controlled clinical trials are needed to define subgroups of patients with higher benefit of the prophylaxis. In individual high-risk patients (size of varices, liver dysfunction, red-colour-signs) beta-blocker prophylaxis may have possible benefit.


[Axonal neuropathy and salazosulfapyridine: slow-acetylator phenotype].

We report a case of an axonal sensorimotor neuropathy involving salazosulphapyridine in a slow-acetylator patient with ulcerative colitis. Rather than hypersensitivity the mechanism of the neuropathy can be assumed to be toxicity. The role played by the respective different metabolites in the occurrence of this uncommon side effect is uncertain.


[Treatment understanding and compliance in asthma of children. Results of a prospective survey].

Compliance with treatment is a crucial factor in the management of asthmatic children which depends on the understanding of the disease and its treatment. The understanding of, and compliance with treatment were evaluated by means of a questionnaire in 50 parents of asthmatic children. 50% of these parents used anti-histamines as maintenance treatment and 30% also used these drugs during attacks. Half the parents knew about the bronchodilator effects of theophylline and B2-agonists. 42% and 30% respectively of the parents thought that the side-effects of theophylline or corticosteroids were few or inexistent; 86% claimed regular attendance to out-patient clinics, but 30% confessed that they had forgotten such drugs as theophylline and antihistamines. 50% took the appropriate therapeutic measures when confronted with a moderate or severe attack of asthma. Each child received 2.3 drugs on average for his or her asthma. This study shows that parents have an insufficient knowledge of asthma treatments, and this may partially explain the poor compliance with therapy as well as the morbidity and mortality associated with childhood asthma.


[Medullary cancer of the thyroid and multiple endocrine neoplastic syndromes].

Despite its rarity, medullary thyroid carcinoma which is hereditary in a quarter of cases and often associated with multiple endocrine neoplasia 2a and 2b syndromes, avises much interest which has lead to the formation of a French multidisciplinary group for the study of calcitonin tumors. This collaboration has resulted in the establishment of a national register and has broadened the knowledge of all specialists concerned. Consequently diagnostic methods are improved, becoming quicker, more reliable and also more thorough due to the setting up of therapeutic protocols. An immunocytochemical staining for calcitonin allows a preoperative diagnosis, providing optimal therapeutic conditions for the patients, especially systematic research of pheochromocytoma and management by a specialized team. Total thyroidectomy and careful lymph node dissection are recommended for all cases. Such progress significantly modifies the prognosis of the disease. Family screening, using plasma calcitonin and, in the near future, genetic studies will hopefully enable the identification of abnormal gene carriers before the clinical stage, thus making total recovery possible.


MHC class II beta-chain expression in the rainbow trout.

A cDNA clone corresponding to the MHC class II beta-chain of the rainbow trout (Oncorhynchus mykiss) has been isolated and used in restriction fragment length polymorphism (RFLP) studies in a family of full siblings of rainbow trout. A very simple RFLP pattern was detected, suggesting segregation of a homozygote AA genotype and a heterozygote AB genotype. The MHC class II beta-chain of the rainbow trout seems to be transcribed in the same type of cells as class II genes of higher vertebrates even though the cDNA clone recognizes atypical messenger sizes of 2.2 kb and 3.6 kb in the analysed family. Surprisingly the transcripts seem to be allele-specific for the assigned genotypes.


Takayasu's arteritis syndrome associated with systemic lupus erythematosus.

Takayasu's arteritis (TA) rarely coexists with systemic lupus erythematosus (SLE). Eighteen cases of TA associated with SLE found by worldwide literature search are reviewed, and an additional unique case is reported in association with the presence of anti-cardiolipin antibody. Patients with TA and/or SLE have similar age of onset and female predominance. The absence of specific SLE markers in patients with TA who subsequently develop SLE suggests that the coexistence of these conditions may be coincidental. The antiphospholipid syndrome in patients with SLE may mimic the occlusive vasculitis of TA.


Tyrosine kinase signal transduction in rheumatoid synovitis.

Explants of synovial cells in rheumatoid arthritis display a transformed phenotype with focus formation and anchorage-independent growth. Many of the cytokines that activate these fibroblasts mediate their action through tyrosine kinase growth factor receptors. Mechanisms of signal transduction via such tyrosine kinases are therefore relevant to the pathogenesis of rheumatoid lesions. Data are presented using the neu oncogene product p185neu as a model system to explore signal transduction by receptor tyrosine kinases. Evidence is shown that increased tyrosine kinase activity in the oncogenic form of this protein may result from dimerization of the tyrosine kinase receptor. In the normal cellular counterpart of p185neu, dimerization appears to be mediated by the action of an as yet unidentified ligand. Dimerization also appears to be important in signal transduction mediated by epidermal growth factor, platelet-derived growth factor, and colony-stimulating factor 1. These cytokines also alter the phenotype of rheumatoid synovial fibroblasts to resemble transformed fibroblasts. Additionally, preliminary data that suggest increased tyrosine kinase activity in rheumatoid arthritis synovia compared with osteoarthritis synovia are presented. Molecular characterization of tyrosine kinase receptors will be an important direction for future studies of the pathogenesis of rheumatoid disease.


Pharmacological characterization of excitatory synaptic potentials in rat basolateral amygdaloid neurons.

The pharmacological properties of synaptic responses in rat basolateral amygdaloid (BLA) neurons were studied using intracellular recording techniques. Three distinct types of synaptic potential were evoked by stimulation of the adjacent ventral endopyriform nucleus: 1) a fast excitatory postsynaptic potential (f-EPSP); 2) a late EPSP (1-EPSP) following the f-EPSP; and 3) a multiphasic hyperpolarization following the initial depolarizing potential. Superfusion of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), a selective non-N-methyl-D-aspartate (non-NMDA) receptor antagonist, blocked the f-EPSP in a concentration-dependent manner. The ED50 for this effect was around 4 microM. In the presence of CNQX, however, a small depolarizing potential remained. This residual depolarizing component was markedly enhanced on removing Mg++ from the perfusing medium and could subsequently be abolished by DL-2-amino-5-phosphonovaleate (DL-APV, 50 microM) indicating its mediation via NMDA receptor-coupled ionophore. The l-EPSP was reversibly blocked by DL-APV. These results suggest that the pyriform cortex-amygdala pathway is mediated through excitatory amino acids acting on non-NMDA as well as NMDA receptors located on the BLA neurons.


Neonatal 6-OHDA lesions differentially affect striatal D1 and D2 receptors.

Lesions to the dopamine (DA) system in early postnatal development have different behavioral consequences compared to lesions made in adulthood. Intrastriatal injections of the neurotoxin 6-hydroxydopamine (6-OHDA) on the day of birth (PO) or postnatal day 1 (P1) produce a selective supersensitivity to D1 receptor agonists and a subsensitivity to D1 antagonists (Neal and Joyce, 1991a). In this paper, we describe the long-term effects of early DA loss on DA receptor regulation. Pups received bilateral intrastriatal injections of the neurotoxin 6-OHDA (4 micrograms per striatum) on PO or P1. Adult rats were killed at 90 days of age and the brains were processed for quantitative autoradiography (QAR) or tyrosine hydroxylase (TH) immunocytochemistry. Cohorts were tested for the behavioral responses to the selective D1 receptor agonist SKF38393 (10 mg/kg). Neonatally lesioned rats exhibited increases in abnormal perioral movements in response to D1 receptor stimulation. There was a heterogenous and patchy loss (40-50%) of [3H]mazindol binding to high-affinity DA uptake sites (a marker of DA terminal density) and a similar loss of TH-like immunoreactivity within the striata of the neonatally lesioned rats. There was also a reduction in the number of mu-opioid receptor patches (labelled with [3H]naloxone), a marker for the striatal patch compartment, and a similar patchy loss of D1 binding sites (labeled with [3H]SCH23390). The binding of [3H]spiroperidol to D2 sites was not altered. This is in contrast to the changes observed following adult 6-OHDA lesions, wherein there is a significant increase in the number of D2 binding sites (Joyce, 1991a,b). The results are discussed with respect to the behavioral consequences of neonatal lesions and the differences between neonatal and adult lesions.


Heart allografts in murine systems. The differential activation of Th2-like effector cells in peripheral tolerance.

Activated CD4+ Th2 cells release cytokines (IL-4,-10) that block activation & cytokine (IL-2/IFN-gamma) release by proinflammatory T (CD4+,CD8+) effector cells. To test the hypothesis that peripheral tolerance to alloantigen is linked to differential activation of CD4+ Th2 cells we measured cytokine transcripts in heart grafts (C57BL/10----C3H/HeJ) and spleens of mice rendered "tolerant" by donor-specific blood transfusion, anti-CD4 mAb pretreatment, and cyclosporine administration. The expression of IL-2/IFN-gamma transcripts was reduced greater than 90% in grafts from tolerant recipients. IL-4/IL-10 transcripts were generally enhanced and persisted in graft and recipient spleen. Accordingly adoptive transfer studies were performed to determine whether Th2-like effectors, which express Fc receptors (FcR), mediate suppression in this model. Unfractionated mononuclear cells (MC) (5 x 10(6), isolated from spleens of heart graft recipients made tolerant by DST, prolonged the survival of test grafts greater than 90 days in irradiated (680 rads) recipients reconstituted with a sufficient number of MC from spleens of naive C3H to precipitate rejection of the test graft in 18.2 days (MST, n = 5). Conversely adoptive transfer of inocula depleted of FcR+ cells on immune complex columns or with anti-FcR mAb 24G2 caused test grafts to be rejected in 8-11 days. These results suggest that peripheral tolerance to alloantigen may be linked to differential activation of Th2 cells that induce anergy by suppression. The possibility that Th2-like effectors mediate peripheral tolerance to self is discussed.


Local immunity in the mammary gland.

The mammary glands of pregnant and non-pregnant sheep were stimulated by infusion of killed Staphylococcus aureus, and the lymphoid cell response delineated with a panel of monoclonal antibodies. Seven days after antigen infusion, the mammary glands of both pregnant and non-pregnant sheep displayed a striking feature, characterised by the presence of numerous CD45R+ MHC class II+ B cells in the periductal connective tissues. These cells were seen to be clustering around blood capillaries with very prominent endothelial cell lining. Some CD5+ CD4+ lymphocytes were scattered among the B-cell clusters, whereas a few CD8+ lymphocytes were seen mainly at the periphery of the B-cell clusters. Fourteen days after antigen infusion, numerous plasma cells were observed, most of them being of the IgA isotype. Seven days after parturition (approximately 40 days after antigen infusion) the number of lymphocytes and plasma cells in the infused glands had declined dramatically. These data indicate that B cell and helper T-cell interaction can take place at the local sites of antigen stimulation in the mammary gland.


[H2-blocker refractory ulcers: what can be done?].

H2-receptor antagonist refractory ulcers are seldom. Less than 5% of duodenal ulcers do not heal under H2-blockers. Today it is more and more accepted that duodenal ulcer is only H2-receptor antagonist refractory after a consequent therapy with H2-receptor antagonists in a standard dose or high doses for at least 2-3 months. The aims of therapy is refractory ulcers include as in non-refractory ulcers healing, pain relief and relapse prophylaxis. Often there is the choice to switch to a more potent antisecretory drug. E.g. ranitidine 300 mg/die was administered successfully in so-called cimetidine resistant ulcers. The specific acid inhibitory profile of omeprazole characterises this substance as a promising alternative in the treatment of H2-receptor antagonist refractory ulcers. In a large controlled clinical therapeutic trial with omeprazole 20 mg/die versus ranitidine 300 mg/die, however, no significant differences were observed in the 2- and 4-weeks healing rates. In two further controlled studies including 99 and 50 patients respectively treated with omeprazole 40 mg/die or cimetidine 800 mg/die and ranitidine 300 mg/die significantly higher healing rates were observed in favour of omeprazole. H2-blocker resistant ulcers are characterised by frequent and rapidly developing relapses. In this cases further treatment with H2-blockers in a two- or threefold daily dose has proved to be effective treatment. The administration of high omeprazole doses (40-60 mg/die) has also produced very convincing clinical results. If the above mentioned therapeutic measure do no provide adequate therapeutic results surgical procedures--usually resection--are indicated.


[Therapeutic possibilities, respective implications in silent ischemia].

In principle, all forms of treatment applied in patients with symptomatic coronary heart disease may likewise be used in silent myocardial ischemia. Based on Bayes' theorem, therapeutic measures may only be applied in patients with a positive exercise ECG with a high likelihood of coronary heart disease, and/or with myocardial ischemia revealed by another, ECG-independent method, such as, for example, thallium-scintigraphy. As symptomatic improvement cannot be expected in patients with silent myocardial ischemia, therapeutic efficacy can only be documented by an improvement in prognosis. Results of controlled randomized trials are not available in silent myocardial ischemia; therapeutic recommendations can, therefore, only be based on analogous results obtained in patients with symptomatic forms of the disease. Apart from reduction of the known risk factors of coronary heart disease, aspirin may be given to all patients at risk. Among the antiischemic antianginal drugs, beta-receptor blocking agents without intrinsic-sympathomimetic activity may be expected to improve prognosis. In asymptomatic patients with left-stem stenosis and three-vessel disease with impaired left-ventricular function (also, in two-vessel disease with a stenotic dominant LAD) aorto-coronary bypass surgery may be considered in order to improve prognosis.


[Breast carcinoma in pregnancy. Clinical, histological and immunohistochemical findings].

The hormonally induced changes in the breast during pregnancy make the diagnosis of breast cancer difficult. Furthermore, examinations during pregnancy tend to concentrate only on the pregnancy itself. In this report the clinical, pathological and immunohistochemical results from 7 patients with breast cancer during pregnancy are presented. All women first noticed the tumor by self-examination. The time periods between discovery of the tumor and medical treatment were between 4 and 22 weeks. An overexpression of c-erbB-2-oncogene coded transmembrane protein p185 could be demonstrated in 4 cases. Of the seven patients, 5 have already passed away. Three of the deceased had p185-positive tumors, and died 4, 8 and 21 months after diagnosis. The two p185-negative patients lived 34 and 65 months post diagnosis. Despite the small amount of cases presented, a trend can be seen that p185 may be an additional prognostic factor for breast cancer in pregnancy.


Rationalizing neuroleptic polypharmacy in chronic schizophrenics: effects of changing to a single depot preparation.

This study investigated the effects of transferring patients on combined depot and oral neuroleptics to a single depot preparation; a secondary objective was to assess the effects of transferring patients from one depot neuroleptic to another. It was found that, whereas transferring from one depot preparation (flupenthixol) to another (fluphenazine) had no clear disadvantage for the patients, changing over from a combined oral and depot (fluphenazine) regimen to equivalent doses of depot alone resulted in an unacceptably high rate of relapse. The reasons for this may relate to either the unique pharmacokinetics of these drugs or subtle qualitative differences between them. It is suggested that caution is necessary whenever attempts are made to rationalize polypharmacy in schizophrenic patients.


Neuroleptic medication and reduced risk of prostate cancer in schizophrenic patients.

A decreased incidence of cancer of the prostate has been demonstrated in a cohort of 6168 chronic schizophrenic patients followed up from 1957 to 1984. A case-control study was performed based on this cohort to determine the possible influence of neuroleptic treatment and other factors on the risk of developing prostate cancer. Thirty-eight male schizophrenic patients who had developed prostate cancer during the observation period were compared with 76 age- and sex-matched controls from the same cohort. The only significant association was that of a reduced risk of prostate cancer among those who had been treated with a cumulative dose of high-dose phenothiazines (primarily chlorpromazine) of 15 g or more. These patients had been treated with an average daily dose of 145 mg chlorpromazine for an average of 12.5 years. No other significant risk factors were identified.


[Collagenous colitis: clinical and histologic improvement after sulfasalazine and topical betamethasone].

We here by report a case of a 42 year-old white woman with an eight-year history of watery diarrhea where the rectal biopsies performed in endoscopically normal mucosa led to the diagnosis of collagenous colitis, characterized histologically by a thickening of the colonic subepithelial basement membrane. A brief review of the current etiopathogenic concepts of this entity is done and the importance of performing rectal biopsies in patients with unexplained diarrhea and normal appearing colon mucosa is stressed. A clinical improvement following therapy with Sulfasalazine and Beta-methasone enemas was found in this patient. We discuss the current views on the therapy and the difficulties of assessing responses to drugs in this condition.


Electroacupuncture enhances activity of adrenal nucleolar organizer regions in ovariectomized rats.

Nucleolar organizer regions (NORs) may reflect the activity of cell differentiation and transcription of nucleolar rDNA; the present paper studied nucleolar organizer regions of the adrenal cortex to explore the regulatory effect of electroacupuncture (EA). Animals were divided into four groups, the ovariectomized electroacupuncture group (OV+EA group, n = 7), the ovariectomized group (OV group, n = 4), the EA group (n = 3) and the control group (CT group, n = 4). Number of AgNORs of 100 cells from zona fasciculata of the adrenal cortex in each case of the four groups was counted at random and the mean+/-SE (number/cell) in each group was calculated as follows: OV+EA group, 2.71 +/- 0.26, OV group, 1.26 +/- 0.15, EA group, 1.21 +/- 0.04 and CT group, 1.48 +/- 0.03. The mean of AgNORs in OV+EA group differed highly significantly from the other three groups (p less than 0.01) as tested with ANOVA and LSD method. No significant difference was found among the other three groups. Gross specimen examination showed that adrenals in the OV+EA group were significantly heavier than those in the other three groups (p less than 0.01). Vaginal smears showed that a response like estrogen-induced one with exfoliative cells appeared in the OV+EA group. EA had no significant effect on normal rats. The results suggest that EA may promote the synthesis and secretion of adrenal steriod hormones, the androgen of which will then be transformed into estrogen in other tissues, thus compensating the deficiency of estrogen induced by ovariectomy.


Osteoelectroacupuncture in the management of vertebrogenic pain syndromes in the lumbar region and lower extremities.

The method for a deep electrical stimulation of the osteofibrous formations involved in a dystrophic process which can be identified according to the criterion of a palpable painfulness or by means of thermography of the projection of local hyperthermia sites is described. An evaluation of the results of the management of vertebrogenic lumbalgia and lumbo-ischialgia by using the proposed method of osteo-electroacupuncture (stimulation by the bipolar asymmetric impulses, O.I msec duration, 5-IO Hz frequency, and up to 200 microA current intensity) has revealed a statistically significant curtailment of the treatment terms and a fewer number of the recurrences when compared to the standard schemes.


Bi-directional transmission of molecular information by photon or electron beams passing in the close vicinity of specific molecules, and its clinical and basic research applications: 1) Diagnosis of humans or animal patients without any direct contact; 2) Light microscopic and electron microscopic localization of neuro-transmitters, heavy metals, Oncogen C-fos (AB2), etc. of intracellular fine structures of normal and abnormal single cells using light or electro-microscopic indirect Bi-Digital O-Ring Test.

In 1985, Omura, Y. discovered that, when specific molecules were placed anywhere in the close vicinity of the path of a light beam (laser), their molecular information, as well as information on electrical & magnetic fields, is transmitted bi-directionally along the path of this light beam. Namely, this information is transmitted in the direction the light beam is projected and towards the direction from which the light beam is coming. This finding was applied to the following clinical and basic research: 1) In the past, using indirect Bi-Digital O-Ring Test, human or animal patients were diagnosed through an intermediate third person holding a good electrical conducting probe, the tip of which was touching the part of the patient to be examined. However, in order to diagnose the patient in isolation from a distance, or a dangerous or unmanagable unanesthesized animal, such as a lion or tiger, the author succeeded in making a diagnosis by replacing the metal conducting probe with a soft laser beam which is held by the one hand of the third person whose index finger is placed in close vicinity of the laser beam generated by a battery-powered penlight-type solid state laser generator. Thus, diagnosis within visible distance, without direct patient contact, became a reality. 2) Using a projection light microscope, by giving indirect Bi-Digital O-Ring Test while contacting with a fine electro-conductive probe on the magnified fine structure of normal and abnormal cells, various normal and abnormal intracellular substances were localized through a third person holding a pure reference control substance with the same hand that is holding the probe as an intermediary for the indirect Bi-Digital O-Ring Test. Instead of the photon beam in a light microscope, the author found that, using an electron beam passing through the close vicinity of specific molecules of specimens in an electron microscope, the molecular information is transmitted to the magnified fluorescent screen, and an indirect Bi-Digital O-Ring Test could be performed through a projected penlight-type solid state soft laser beam on the magnified intracellular structure through an observation glass window. Using the magnified fine structure of the cells, by either a light projection microscopic field or electron microscope, in various cancer cells of both humans and animals, Oncogen C-fos (AB2) and mercury were found inside of the nucleus. Integrin alpha 5 beta 1 was found on cell membranes and nuclear cell membranes of cancer cells. Acetylcholine was not found anywhere within cancer cells.(ABSTRACT TRUNCATED AT 400 WORDS)


The origin of the five elements in the traditional theorem of acupuncture: a preliminary brief historic enquiry.

Five Xing is an important integral in the traditional theoretic basis of acupuncture and traditional Chinese medicine. The word Xing has been translated as Element. However, it actually denotes movement and activity. The word element implies a stationary state. Some of the evidence in ancient Chinese literature was reviewed to support the hypothesis that Five Xing were originally meant to be the Five Xing Xin (Moving Stars, i.e., Planets). By the 4th century B.C., associations of the Stars with human events gradually evolved. However, between the 4th and the 6th century A.D., when the Taoist scholar-physicians expanded the Five Xing into abstractive concepts, they used the five basic materials as their representatives. Since they were basically alchemists and not astronomers, they apparently minimized the relationship between the Five Moving Stars and the human illnesses. It is, therefore, proposed that the usage of the word Element be discontinued and the word Xing be employed as is.


[Three-dimensional computerized tomography in trauma surgery. A case presentation].

Three-dimensional images of bone structures can easily be reconstructed from computed tomography data. The technique and the advantages of contemplating reconstructions of bone defects in different directions are described. A special software programme allows to combine all data which are given by the standard CT. The slice diameter is two millimetres. The reconstruction shows bone surfaces in all required views. In traumatic and orthopaedic surgery 3-D-CT is useful in the analysis, detection and preoperative planning of comminuted bone injuries of the spine, pelvis, knee, shoulder and the calcaneus. With this technique it is possible to see the bone in its topographic constellation, to observe fractures in unusual directions and to identify fragments from each other. The surgeon gets a lot of additional information which is helpful to decide on the incision, the reduction and the fixation of fragments. Undesirable effects during operation are reduced, which leads to a more accurate treatment and subsequent better results.


[Limits for recognizing linear fractures of the cranial vault in radiologic diagnosis].

In case of failure to detect a vault fracture in fatal head injury the physician is often accused of medical negligence. The material for the study consisted of a macerated vault with fractures in different widths. In order to detect the threshold of perceptibility it underwent radiographic examination taking pictures under different projections. Our finding is that the presence or absence of a radiographic fracture depends on the width and direction of the fracture. Therefore it is impossible to detect every single linear fracture of the vault in routine skull examination. Clinical and medico-legal consequences are discussed.


[Stabilization of the injured shoulder joint with PDS cord].

A successful treatment of the acromioclavicular separation is the repair of the acromioclavicular and coracoclavicular (CC-)ligaments and a stable reduction of the acromioclavicular (AC-)joint. To avoid dangerous breakage and migration of the K-wire an abduction humeral splint is necessary immobilizing the injured shoulder for 5-6 weeks. In the years 1987-1989 40 patients suffering AC-separation were treated (34 Tossy III separations, 4 Tossy II separations, 2 Tossy I separations). In these cases a stable reduction was achieved by a transarticular K-wire fixation and a combination of AC- and CC-fixation by loops. In 1987 wire loop was used. In 1988 a combination of wire and Polydioxanon (PDS) loops was used. The PDS-loop, a slowly resorbable suture material, fixed the CC-ligament. In 1989 the AC-joint was stabilized by a PDS-loop as well. The examination of 31 patients 6-24 months after the operation showed good clinical results no matter whether PDS-loops or wire loops were used. The advantage of the transarticular K-wire fixation in combination with PDS-loops was the easy removal, which could be done in mostly of the cases as an outpatient procedure. An operation of the AC-Tossy III separation on patients beyond their 4. decade should be well considered. A long time of treatment, remaining pain and a limitation of shoulder movement must be expected.


[Drill wire osteosynthesis in subcapital humerus fractures: percutaneous or open procedure?].

Between 1978 and mid-1990, 135 patients suffering from dislocated, non-luxated fracture of the humerus at the anatomical neck (fractura colli anatomici) were treated by means of open or closed percutaneous drill wire osteosynthesis. Follow-up examination after an average of 9 months did not show any significant differences between the two surgical approaches in 117 patients, independent of the shape of the fracture. However, in about 30% of the cases it was impossible to employ the percutaneous approach due to the presence of an obstacle to reduction, so that open reduction and fixation was the only choice. A great majority of the functional results must be considered as good, fractures of the tubercles having the most unfavourable prognosis independent of the surgical technique. It is, therefore, recommended to first try closed reduction with percutaneous drill wire osteosynthesis. If there are any obstacles to reduction, open reduction should be restored to during the surgery session.


[Treatment of osteomyelitis and reconstructive measures in patients with war injuries].

54 wartime trauma patients injured by bombs, shell splinters or rockets were treated between 1985 and 1989 in the Orthopaedic Clinic of RWTH Aachen. Lesions of the lower limbs were dominating (78%). Treatment was often tedious, and up to 12 operations had to be performed. In 81% one extremity was injured, in 17% two extremities were involved. 78% of the patients were already primarily treated at home, mostly by amputations. 33% of the patients suffered from bone infections at admission. Infections were mostly caused by Staphylococcus aureus or Pseudomonas aeruginosa. In 41% of all patients operative treatment for osteomyelitis was necessary (37% sequesterectomies, 44% stabilisations with fixateur externe). In advanced bone infection amputations were indispensable in 28%. In 27% of our wartime trauma patients reconstructive surgery was performed (spongiosa transplantations in 91%, stabilisation with fixateur externe in 44%). In none of our patients treated with reconstructive surgery an amputation was necessary later on.


[Early management of a fracture of the coxal end of the femur in elderly patients].

The demographical structure of the Federal Republic of Germany leads to an increasing number of very old patients in traumatology. Even minor accidents can cause severe injuries including operative treatment. In this retrospective study the data of 219 patients older than 70 years who had to undergo an operation at the femoral neck region are analyzed. The aim of operative management was a very early operation and a fixation device, that allows an immediate full weight-bearing. Fractures of the femoral neck are stabilized by hemiarthroplasty, fractures of the trochanteric region with the dynamic hip screw. 90% of our patients could be operated within 24 hours, the mortality rate during the first 14 days was 3%.


[Capsular ligament lesions of the knee joint. 5-9 year results of surgical management].

5-9 years follow-up results following 26 out of 37 operated injuries of the ligaments of the knee are reported. While 18 patients described the result of the operation as very good and good, the objective findings presented only 5 stable knees.


[Orofacial injuries in skateboard accidents].

In a clinical study, 25 accidents involving injuries by a fall with a skateboard were investigated and classified in respect of epidemiology, accident mechanism and injury patterns in the facial region. Accident victims are predominantly boys between 7 and 9 years of age. A multiple trauma involving the teeth and the dental system in general and the soft parts of the face is defined as a characteristic orofacial injury pattern in skateboard accidents. The high proportion of damage to the front teeth poses problems of functional and aesthetic rehabilitation necessitating long-term treatment courses in children and adolescents. Effective prevention of facial injuries may be possible by evolving better facial protection systems and by creating areas of playgrounds where skateboarders can practise safely.


A review of the literature on the use of ultrasonography in schistosomiasis with special reference to its use in field studies. 1. Schistosoma haematobium.

This review presents an outline of the pathology resulting from Schistosoma haematobium infections, and the ways in which the lesions can be investigated. The use of ultrasonography is covered in detail. Ultrasonography can provide direct information about lesions in internal organs, and thus provide information about patterns of morbidity and about the regression of pathological changes after treatment. The method has the advantages that it is non-invasive, and is also relatively inexpensive and can be used under field conditions. Ultrasonography has already been used in a number of epidemiological studies in areas where S. haematobium is endemic. The method has proved to be feasible and useful. However, the methodology used for ultrasound studies has varied considerably, so that it is difficult to make valid comparisons between results obtained in different places or at different times. A standardized methodology for making observations and recording the results is needed if the full potential benefit of using ultrasound in the monitoring of schistosomiasis control projects is to be realised. The correlation of results obtained using ultrasound with the results of clinical, parasitological and other observations has been investigated in a number of studies, but many questions remain to be answered.


A review of the literature on the use of ultrasonography in schistosomiasis with special reference to its use in field studies. 2. Schistosoma mansoni.

This review presents an outline of the pathology resulting from Schistosoma mansoni infections, and the ways in which it can be investigated. The use of ultrasonography is covered in detail. Ultrasonography can provide direct information about lesions in internal organs, and thus provide information about patterns of morbidity and about the regression of pathological changes after treatment. The method is non-invasive, and can be used under field conditions. Ultrasonography is valuable for the study of hepatosplenic pathology, to detect lesions such as the development of periportal fibrosis and the enlargement of the portal vein, which can indicate the development of portal hypertension. This may lead to a severe outcome, including bleeding from oesophageal varices, which is a principal cause of death from S. mansoni infection. A problem with the use of ultrasonography is that the mild lesions likely to be observed in population surveys are not always easy to assess. Ultrasonography has already been used in a number of epidemiological studies of S. mansoni infection, and has proved to be feasible and useful. However, the methodology used for ultrasound studies has varied considerably, so that it is difficult to make valid comparisons between results obtained in different places or at different times. A standardized methodology for making observations and recording the results is needed if the full potential benefit of using ultrasound in the monitoring of schistosomiasis control projects is to be realised.


A review of the literature on the use of ultrasonography in schistosomiasis with special reference to its use in field studies. 3. Schistosoma japonicum.

This paper gives a brief description of the pathology resulting from Schistosoma japonicum infection, and ways in which it can be investigated. It then reviews reports of the application of ultrasound in investigating lesions in schistosomiasis japonica, including papers published in Chinese and Japanese. Ultrasonography has been widely used for the diagnosis of schistosomiasis and for the investigation of pathological changes resulting from the infection. Marked and characteristic changes are observed in the structure of the liver parenchyma in advanced disease. Chronic pathology may be seen as a result of past infection. Animal studies have been used to compare ultrasound images with actual pathological changes. Ultrasonography can also be used to detect early changes, for example periportal fibrosis, which can indicate the development of portal hypertension. The problem of differential diagnosis is discussed.


Studies on ultrasonographic diagnosis of schistosomiasis japonica in China--a review of selected Chinese studies.

Diagnostic ultrasound has been used in China since the early 1960's in the investigation of Schistosoma japonicum infection. This paper presents an overview of selected papers from the Chinese literature on the use of ultrasound in studies of schistosomiasis japonica, and briefly reports the results of studies not yet published elsewhere. The method is compared with other diagnostic procedures, and the benefits and limitations of using ultrasound are discussed.


The use of diagnostic ultrasound in schistosomiasis--attempts at standardization of methodology. Cairo Working Group.

This paper summarises the conclusions of a workshop held in Cairo in 1990 to discuss the standardization of the use of diagnostic ultrasound in schistosomiasis. For epidemiological purposes, it is very important that ultrasound examinations should be carried out and recorded in a standardized way, in order to ensure that results obtained in different places and at different times can be compared. The workshop did not attempt to produce final recommendations, but it did make tentative proposals in a form suitable for field testing. The paper discusses the general problems involved in carrying out and recording ultrasound examinations in a standardized way. The special points involved in assessing lesions due to the three main forms of schistosomiasis are then considered in detail. Methodology that can be used in epidemiological work with large numbers of people is emphasised.


Liver ultrasound findings in a low prevalence area of S. japonicum in China: comparison with history, physical examination, parasitological and serological results.

The paper describes a study carried out in a community in Dongdian township, Anhui Province, People's Republic of China. Medical history and the results of a physical examination, ultrasound investigation, parasitological and serological tests for Schistosoma japonicum infection were compared in 661 persons of 169 households. A lack of correlation between parasitological and serological indicators of infection and morbidity was observed in this area of low (6.4%) prevalence and intensity of infection. The prevalence of abnormal ultrasound findings in the liver in this population was high (56%), and was significantly higher than the prevalence of S. japonicum infection. The abnormal ultrasound findings correlated with a history of schistosomiasis, and the correlation increased significantly according to the number of times treated and the time since the last treatment, which suggested that past parenteral treatment has a role in the high rate of abnormal liver ultrasound findings. The significant correlation between the qualitative and quantitative serological results and abnormal ultrasound parenchymal patterns suggests that cross-reactivity between the etiology of the parenchymal disease and these tests is occurring. The presence of HBsAg correlated with the composite presence of ultrasound abnormalities of the liver parenchyma: increased echogenicity, periportal fibrosis and/or nodules and irregular fibrosis, whereas a normal ultrasound pattern was associated with the absence of HBV antigenemia.


Inter-observer variance in ultrasonographical assessment of Schistosoma mansoni-related morbidity in young schoolchildren.

49 Sudanese schoolchildren aged 6-9 years with Schistosoma mansoni infection were ultrasonographically examined by two independent observers in a double-blind fashion. The first observer recorded normal appearance of the liver in 23 cases, whereas the second observer recorded the appearance as normal in 33 cases. There were 23 concordant observations. For Grade I periportal fibrosis (PF), 13 observations were concordant. PF Grade II was rarely observed (2 versus 3 cases), and Grade III was not recorded at all. In total, 38 out of 49 observations were concordant (77.5%). These preliminary data from two ultrasound observers, from observations on a limited number of patients, can be seen as an indication of a potential inter-observer variation of around 20% for the distinction between the absence of PF and a low level of PF.


Ultrasound in schistosomiasis--a critical look at methodological issues and potential applications.

This is the concluding paper of a series on the use of diagnostic ultrasound in the investigation of schistosomiasis. An earlier chapter in the volume discussed standardization of the methodology, and of recording, when ultrasound is used for epidemiological purposes. The present paper discusses some other requirements for obtaining ultrasound data which can be used to make valid comparisons within and between studies. Since there is an inherent variability in the interpretation of results from ultrasound images, quality control and the training of observers are both essential. It is also necessary to collect more information for each endemic setting about possible concomitant diseases which might lead to misinterpretation of results. Furthermore, the analysis of the data obtained must be uniform if valid comparisons are to be made. A final section considers applications of ultrasonography in research and control programmes. The technique should make it possible to obtain a better understanding of the extent and distribution of organ damage due to schistosomal infection in different geographical areas, and of the way in which lesions develop over time, or may regress in response to treatment. Since ultrasonography will always remain a relatively labour-intensive and expensive technique, it is necessary to establish, in different settings, how its findings correlate with the results of parasitological, serological and biochemical tests. The ultimate aim is to build up a body of information on the potential of ultrasonography, in combination with other procedures, in the various possible approaches to morbidity control.


Dehydroepiandrosterone: antiglucocorticoid action in mice.

The acute effect of dehydroepiandrosterone (DHEA) and its conjugate, DHEA-sulfate (DHEA-S) on glucocorticoid action was tested in vivo using male Swiss-Webster mice. The authors found that DHEA and DHEA-S significantly inhibited induction of hepatic tyrosine aminotransferase activity, although the former was more potent. This inhibition was dose- and time-dependent and was not demonstrable with other steroids. The same inhibitory effect of DHEA was seen with kidney tyrosine aminotransferase induction, as well as with liver and kidney ornithine decarboxylase enzyme activity, another glucocorticoid-induced enzyme. The conclusion is that DHEA acts acutely as an antiglucocorticoid and exerts its effect in different glucocorticoid-sensitive systems.


On some technical aspects of direct DNA diagnosis of the fragile X syndrome.

Direct DNA analysis of fragile X [Fra(X)] mutations has already shown its clear superiority for postnatal and prenatal diagnosis of the disorder and for carrier detection. However, it is of great importance to have conditions which guarantee optimal reliability and sensitivity. Some mutations may be more difficult to detect, especially in female carriers: this is the case for small amplifications of the CGG repeat (premutations) or for smears which can be generated by the instability of the full mutation in somatic tissues. We present the various alternatives (probe/enzymes combinations) for Southern blot based diagnosis, the possible artefacts and our detailed experimental protocol, which has given excellent results on a large number of families. While detection of amplification, using for instance EcoRI, appears sufficient for initial testing of mentally retarded patients, once the fra(X) diagnosis has been established, we favor the use of an EcoRI+EagI digest, which detects both amplification and abnormal methylation, for analysis of the family, including carrier detection and prenatal diagnosis. We discuss the place of proposed PCR based techniques for detection of mutations, or for indirect tracking using polymorphic microsatellites in the immediate vicinity of the fra(X) locus.


Characterization of new PCR based markers for mapping and diagnosis: AC dinucleotide repeat markers at the DXS237 (GMGX9) and DXS102 (cX38.1) loci.

Genomic insert DNAs from 45 probes representing 113.4 kb of the X chromosome were screened for AC dinucleotide repeat sequence. Two new AC repeat sequences were identified with length polymorphism based on variation in repeat copy number. One at DXS237 exhibits 44% heterozygosity and is potentially useful for rapid diagnosis and mapping of X-linked disorders in Xp22.3. The other, at DXS102 in Xq26, has 71% heterozygosity. This marker will improve accuracy of diagnoses by linkage for families with Börjeson-Forssman-Lehmann syndrome. Review of the literature has identified 31 PCR based markers on the X chromosome, with minimum heterozygosity of 50%, applicable to the mapping and diagnosis of X-linked disorders.


Carrier detection of the fragile X syndrome with flanking RFLP markers and linkage analysis.

Linkage analysis was performed in 34 fragile X (fra(X)) families in order to study the efficiency of carrier detection using the restriction fragment length polymorphisms (RFLPs) closely linked to fra(X) locus (FRAXA). The marker loci used were F9, DXS105, DXS98, DXS369, DXS297 and DXS477 proximally and DXS465, DXS296, DXS304, DXS52 and F8C distally to FRAXA. Flanking heterozygosity was achieved in 60% of the females with a combination of 3 restriction enzymes and 6 closest RFLP markers. When adding more distant markers and other restriction enzymes to the analysis, the proportion of females heterozygous for flanking polymorphisms increased to 96%. With RFLP-analysis most (85/91) females at high risk of being a carrier could be separated clearly into 2 groups: those with a very low and those with a very high risk. The 6 cases with a recombination between flanking markers did not benefit from RFLP-analysis.


Direct DNA analysis of fragile X syndrome in Spanish pedigrees.

Eleven complete Spanish pedigrees with fragile X syndrome were analysed by Southern blotting with the DNA probe StB12.3 previously isolated and described by Oberlé et al. [1991]. This probe allowed the direct detection of affected males and carrier females and was able to distinguish between normal males and normal transmitting males (NTMs). One hundred and twenty three individuals were analyzed, 115 from the pedigrees and 8 from the general population. Five mosaic cases were found (4 males and one female) showing both the premutation and the full mutation. One half of the females with the full mutation were mentally retarded but no female with mental retardation carried the premutated pattern, suggesting that the absence of the full mutation in females is a very good criterion for pre-or postnatal diagnosis of normal mental status.


Dose-related effects of doxazosin on plasma lipids and aortic fatty streak formation in the hypercholesterolemic hamster model.

Doxazosin, an alpha 1-adrenergic inhibitor, has been shown to decrease hypertension and plasma lipids, especially total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C), thus reducing certain risk factors associated with increased incidence of cardiovascular disease. One preliminary report indicated that the decrease in LDL-C in hypercholesterolemic hamsters treated with doxazosin was associated with a reduction in fatty streak formation. However, since the effects of doxazosin on plasma lipids, aortic fatty streak development, or the relationship between the two have not been studied in a dose-dependent manner, these effects were further investigated over varying doses of doxazosin (0, 1, 5, 10, and 20 mg/kg body wt/day) during a 10-week period. Doxazosin administration was associated with a dose-dependent decrease in LDL-C of 2%, 29%, 52%, and 60%, whereas the degree of fatty streak formation was reduced 11%, 45%, 76%, and 92% compared with controls, with the first statistically significant decrease for both parameters at the 10 mg/kg dose. Significant correlations between LDL-C concentrations and fatty streak area suggest that doxazosin altered aortic lipid infiltration primarily by its effect on plasma lipids. However, the 20 mg/kg dose of doxazosin significantly decreased lesion area compared with the 10 mg/kg dose without a further effect on plasma lipid concentrations. Three animals at these higher doses demonstrated no stainable lipid inclusions while maintaining plasma lipid values similar to their cohorts. These exceptions to the lipid-lesion relationship raise the possibility of additional effects of doxazosin, which may occur independent of or in concert with lipoprotein cholesterol lowering, on lesion formation.


Abnormal isoform of prion proteins accumulates in the synaptic structures of the central nervous system in patients with Creutzfeldt-Jakob disease.

A new method, which enabled the first immunohistochemical documentation of abnormal prion protein (PrP) in all patients with Creutzfeldt-Jakob disease (CJD), was established. This method designated as "hydrolytic autoclaving" revealed punctate PrPCJD stainings around the neuronal cell bodies and dendrites in CJD brains. These punctate stainings were almost identical with that of synaptophysin, suggesting PrPCJD accumulations in the synaptic structures. Subcellular fractionation revealed that prion protein in Creutzfeldt-Jakob disease (PrPCJD) was most concentrated in the synaptosomal fraction. In CJD patients with a long clinical course, synaptophysin immunoreactivity decreased, and synaptic PrPCJD accumulated with a wider distribution. These results suggest that synaptic PrPCJD accumulations might be responsible for the neuronal dysfunction and degeneration in CJD.


Linkage and risk assessment in fragile X families using new DNA probes at Xq27.

Until recently few polymorphic loci had been genetically mapped close to the fragile X syndrome locus [FRAXA]. Six polymorphic loci, DXS369, DXS297, DXS296, DXS304, IDS and DXS374, have now been mapped closer to the fragile X FRAXA than in the present study. We report the results of genetic linkage analysis of 32 fragile X [fra(X)] families using 12 polymorphic loci including these new markers. Cytogenetic and molecular data were combined in two-point linkage analysis for the estimation of lod scores and carrier probabilities in potential carriers. Combined with results from previous studies, recombination fractions (0) corresponding to the maximum lod scores (Z max) were obtained for each of the 12 loci versus FRAXA. Recombination fractions between marker loci in the families were also calculated. The data were evaluated to determine the efficacy of using the strategy suggested by Suthers et al. (1991a) for molecular studies in fra(X) families. The large proportion of females heterozygous for at least one locus (83%) and of females heterozygous for flanking loci (60%) indicate that this is a very useful diagnostic strategy. Use of these new marker loci substantially changed the carrier risk estimates for members of 7 of the 32 families from the risk estimates previously calculated on the basis of less closely linked probes available prior to 1989.


HUT 78 T cells bind to noncytokine-stimulated keratinocytes using a non-CD18-dependent adhesion pathway.

The initial in vitro observation that cultured keratinocytes, when treated with cytokines such as gamma interferon, increased the binding of T lymphocytes, opened up a whole new avenue of research to understand epidermal trafficking patterns in inflammatory skin diseases. A growing body of data strongly supports the in vivo role of lymphocyte-function-associated antigen-1 (CD18) expression by T cells in the binding to intercellular adhesion molecule-1 (CD54) expressing keratinocytes. To further explore the molecular basis for other possible adhesive interactions involving T cells and skin-derived cellular constituents, the authors used 2 cell lines (HUT 78 cells and Jurkat cells) and added them to multipassaged human keratinocytes, fibroblasts, and melanocytes. The skin-derived cells were treated with cytokines alone, or in combination, with a phorbol ester. HUT cells were capable of binding to keratinocytes in the absence of pretreatment with cytokines at 25 degrees C, which was not inhibited by anti-CD18 antibodies, or sensitive to reducing the temperature of the adhesion assay to 7 degrees C. Fibroblasts and melanocytes also constitutively bound HUT cells, but the binding to fibroblasts was highly temperature-sensitive. When keratinocytes were pretreated for 48 hours with gamma interferon plus phorbol ester, a "superadhesive" state was induced, resulting in a synergistically increased binding ability of both HUT cells and Jurkat cells. This effect was related to quantitative increases in keratinocyte intercellular adhesion molecule-1 expression. Several other clear-cut qualitative and quantitative differences were detectable in the ability of HUT cells and JS cells to bind to nontreated and cytokine/phorbol ester-treated keratinocytes, fibroblasts, and melanocytes. These results emphasize the complexity of molecular associations underlying T-cell trafficking patterns, potentially operative in the dermal and epidermal compartments of the skin.


Thrombospondin and other possible related matrix proteins in malignant and benign breast disease. An immunohistochemical study.

Thrombospondin (TSP), which plays an important hemostatic role, is a 450-kd cytoadhesive protein present in the alpha granules of platelets. In vitro experiments using cultured malignant cells suggest that it may help to mediate malignant cell attachment to extracellular matrix and may be important in cancer invasiveness. The authors studied a group of malignant and benign breast tissues for the expression of TSP and provide evidence that TSP may have a role in tumor invasiveness. Using immunohistochemical techniques, the authors found in 48 fresh specimens of breast carcinoma that TSP stained strongly in the desmoplastic stroma or at the basement membrane associated with the malignant ductal epithelium. Tumor cells abutting these tissues revealed cytoplasmic staining for TSP. Stronger TSP staining was seen in the poorly differentiated tumors. These findings were compared with those of normal and benign breast tissue, which showed no TSP staining apart from reactivity in the large distended cysts of fibrocystic disease and faint staining in the stroma of fibroadenomas. Staining for integrin was positive in lymphocytes of both malignant and benign breast disease and equivocally also in the stromal cells of the breast cancer tissue. Immunoreactivity to TSP in tissues was also compared with that of fibronectin, laminin, and collagen type I, III, and IV. The overall findings suggest that thrombospondin may have a role in the tumor biology of invasiveness.


Immunocytochemistry of paraneurons in the female urethra of the horse, cattle, sheep, and pig.

The aim of this study is to describe the presence of neuroendocrine (NE) cells (paraneurons), producing biogenic amines and/or peptidergic hormones, in the female urethra of cattle, sheep, pigs, and horses, by means of histochemical and double labeling immunofluorescent techniques. 5-Hydroxy-tryptamine-, chromogranin A-, cholecystokinin- and somatostatin-containing NE cells are present in the urethral epithelium of all the species studied, with the unique exception of the lack of somatostatin cells in the horse. Paraneurons containing 5-hydroxytryptamine colocalized with chromogranin A or cholecystokinin were also found in all subjects. Such active substances are hypothesized to play a role in the contraction of the urethral musculature, emission of urogenital fluids, and inhibition of endocrine and exocrine secretions.


Low-dose trimethoprim-sulfamethoxazole prophylaxis for toxoplasmic encephalitis in patients with AIDS.

To determine the efficacy of low-dose trimethoprim-sulfamethoxazole (trimethoprim, 160 mg plus sulfamethoxazole, 800 mg; one tablet twice daily, 2 days per week) as primary prophylaxis against toxoplasmic encephalitis in patients with human immunodeficiency virus (HIV) infection and previous Pneumocystis carinii pneumonia. A retrospective study. Tertiary referral teaching hospital. During a 3-year period after primary episodes of P. carinii pneumonia, 60 patients received trimethoprim-sulfamethoxazole, and 95 patients received pentamidine (aerosolized in 78 patients and intravenous in 17 patients) as secondary prophylaxis. No patient in the trimethoprim-sulfamethoxazole group and no patient seronegative for Toxoplasma gondii developed toxoplasmic encephalitis, compared with 12 of 36 (33%; 95% Cl, 19% to 51%) seropositive patients in the pentamidine group (trimethoprim-sulfamethoxazole compared with pentamidine, P = 0.008). A significant difference was seen in the time to development of toxoplasmic encephalitis between the trimethoprim-sulfamethoxazole group (no case at 1153 days) and the pentamidine group (median time, 460 days) (P = 0.004). Neither the CD4+ lymphocyte count at the start of prophylaxis nor zidovudine therapy during the period of prophylaxis influenced the rate of toxoplasmic encephalitis in any group. Low-dose trimethoprim-sulfamethoxazole (four tablets per week) appears to be effective prophylaxis against toxoplasmic encephalitis in HIV-infected patients with previous P. carinii pneumonia. A prospective, randomized, controlled study is needed to further evaluate these findings.


Anticoccidial activities of 7-bromo-N-(2-imidazolidinylidene)-1H-indazol-6-amine and other alpha 2 adrenergic agonists.

Activity against the coccidial pathogen Eimeria tenella in chickens has been discovered among alpha 2 adrenergic agonists. The clonidine analog 7-bromo-N-(2-imidazolidinylidene)-1H-indazol-6-amine was active in feed at 7.5 ppm, a concentration similar to the use levels of potent commercial agents, e.g., maduramicin. Additional alpha 2 agonists were also found to have anticoccidial activity, for example, the catecholamine nordefrin, which is chemically unrelated to clonidine. However, alpha 1 agonists and alpha antagonists were inactive. These observations imply that anticoccidial effects reflect involvement of a receptor with the characteristics of the vertebrate alpha 2 adrenoceptor. alpha 2 agonists that permeate the blood-brain barrier (like clonidine) inhibit feed intake at efficacious levels, whereas those that are restricted to the peripheral compartment (such as catecholamines) do not inhibit feed intake as much. Hence, anticoccidial efficacy may be a peripheral adrenergic effect whereas depression of feed intake is likely centrally mediated.


Further investigation of anticoccidial activity of 7-bromo-N-(2-imidazolidinylidene)-1H-indazol-6-amine.

The clonidine analog 7-bromo-N-(2-imidazolidinylidene)-1H-indazol-6-amine exhibits potent activity against Eimeria tenella infections in chickens. Disease control was abrogated by a selective alpha 2 antagonist, which is consistent with the dependence of such activity upon binding to receptors with characteristics of the vertebrate alpha 2 adrenoceptor. Lack of significant activity against the parasite in tissue culture and our inability to detect significant binding of alpha 2 adrenergic ligands to E. tenella imply that the anticoccidial action may be an indirect effect mediated by the host. Efficacy varied, depending upon the Eimeria species, being greatest for the cecal species E. tenella and less for the intestinal species. The effects described differ substantially from previous accounts of adrenergic actions on parasitic protozoa. The evidence suggests that we have observed a new mechanism of action for antiparasitic drugs.


Enhancement of gamma-glutamylcysteine synthetase mRNA in rat kidney by methyl mercury.

Glutathione (GSH), a major cellular antioxidant, is elevated 2- to 3-fold in kidneys of rats during prolonged treatment with mercury as methyl mercury hydroxide (MMH). Increased renal GSH is accompanied by a dose- and time-related elevation in the relative abundance of mRNA hybridizable to a cDNA probe which encodes renal gamma-glutamylcysteine synthetase (GCS), the rate-limiting enzyme in GSH synthesis. Renal GCS mRNA is maximally elevated 4.4-fold at 3 weeks following initiation of MMH treatment. Enhancement of GSH and GCS mRNA content corresponds to a relative sparing of renal cells from oxidative tissue damage during MMH exposure. These observations suggest that increased synthesis of GSH at the genetic level occurs as an initial adaptive response to mercury-induced oxidative stress in kidney cells.


[Progress in oncogene and antioncogene research].

Recent progress in molecular biology of cancer revealed that mutations which potentiate the activities of proto-oncogenes create the oncogenes to force the growth of tumor cells. On the other hand, inactivation of antioncogenes results in the generations of tumor cells. The progression of many tumors to full malignancy requires both types of changes of the tumor cell genome.


Characterization of field isolates of suid herpesvirus 1 (Aujeszky's disease virus) as derivatives of attenuated vaccine strains.

Field isolates of suid herpesvirus 1 (Aujeszky's disease virus) from Poland and Hungary were identified by restriction fragment pattern analysis as derivatives of attenuated vaccine strains. The Polish isolates were found to be related to the BUK-TK-900 strain (Suivac A) which is widely used as a live vaccine in Poland, and the Hungarian isolates were related to the Bartha K-61 vaccine strain widely used in Hungary. Pigs experimentally infected with derivatives of BUK-TK-900 or BUK-TK-900 itself were found to develop gI-antibodies, while pigs infected with derivatives of Bartha K-61 showed a gI-negative response.


[Plasma neuropeptides in affective and anxiety disorders].

The levels of Neurotensin, VIP, Somatostatin, beta-endorphin and Bombesin have been investigated in plasma of 16 depressed and 20 anxious patients. VIP and Neurotensin were found significantly decreased in patients vs a group of 20 controls. Neurotensin levels returned to normal values after recovery. There were no significant differences from the normal in the concentrations of Somatostatin, beta-endorphin and Bombesin in the disease groups.


Structure-function studies of HIV-1: influence of long terminal repeat U3 region sequences on virus production.

DNA sequence analyses of several human immunodeficiency virus (HIV) isolates revealed extensive genetic diversity in the env gene and, to a lesser extent, in other regions of the viral genome, including the long terminal repeat (LTR) sequences. Since the LTRs contain elements responsible for the control of transcription, the difference in the LTR region may play a crucial role in the overall replication rate of HIV. To evaluate the role of the LTR, we have constructed a number of infectious hybrid HIV molecular clones containing LTRs from different proviral DNAs linked to the body of the viral genome, and analyzed them in a transient expression system. Both parental and hybrid proviral DNAs were transfected into human rhabdomyosarcoma cells for monitoring virus production. Proviral DNA designate pZ6 (HIVZr6) showed a high level of virus in the medium of the transfected culture in comparison to the pHXB2 (HIVHTLV-III) and pARV (HIVSF-2) DNAs. Hybrid proviral DNAs containing viral genes from pZ6, linked to LTR U3 sequences of pHXB2 and pARV at the 5' end, showed virus production similar to the levels observed with pZ6. These results indicate that the extent of virus production does not correlate with the LTR U3 sequences, and may involve other regions of the viral genome.


Cellular aspects of early T-cell development.

Although the nature of the precursor cells seeding the thymus is still uncertain, their immediate progeny in the adult murine thymus have now been isolated. These lymphoid-restricted, prothymocyte-like cells express CD4, but neither CD4 nor CD8 seem to be involved in the early steps of T-cell development. Cytokines produced by stromal cells are likely to be involved in intrathymic T-cell development, but interleukin-2 and interleukin-4 do not appear to be required. There is still no satisfactory cell-culture model of intrathymic T-cell development. Current culture systems reflect only fragments of the process, or are models of extrathymic developmental pathways.


Thymic selection.

The ability of T cells to recognize foreign antigens, distinguish them from self antigens, and regulate immune responses depends largely on which of the vast array of different T-cell receptors they display on their surface, the so called T-cell repertoire. Two selection processes, positive and negative selection, operate on developing T cells and allow only a subset of all possible T cells to differentiate. New developments in transgenic mouse technology and the study of T-cell signalling are unravelling these fascinating processes.


Extrathymic T-cell selection.

The T-cell repertoire is formed during T lymphocyte maturation in the thymus. There is, however, increasing evidence that there are additions to, and modifications of, this repertoire by extrathymic events. Most importantly, mature peripheral T cells are not only susceptible to activation signals but also to tolerance induction upon encounter of extrathymic antigen. An understanding of these inactivation processes should result in strategies for specific immunosuppression in organ transplantation and autoimmune diseases.


[Liver enzyme values (gamma-GT, GOT, GPT) in intoxicated drivers at the time of the offense].

The enzyme activities (GGT, GPT, GOT), especially of the gamma-glutamyl-transpeptidase, is considered to be a sign of alcohol abuse. Of 219 blood samples taken from drunken drivers for blood alcohol determination, 81 (37.6%) exhibited pathological enzyme activities. In 55.6% of them specifically pathological gamma-Gt values were found. In the cases with normal enzyme activities the average value of blood alcohol concentration was 1.61%, the pathological cases had 1.81%. In the latter cases more than 50% of the drunken drivers involved were workers. Most of them were 30-49 years old (45%), but also young drivers up to 21 years old were represented (about 25%). There was also a difference between the group with normal and pathological enzyme activity concerning their drinking behaviour. The group with pathological values occurred during the week, the other group was mostly arrested at the weekends. Of the total, 6.8% (15) were women, 4 exhibited pathological enzyme activities; three of them were 21-29 years, one was 36 years old. The data obtained demonstrate that drunken drivers exhibit a tendency to alcohol abuse.


Domains of cellular retinoic acid-binding protein I (CRABP I) expression in the hindbrain and neural crest of the mouse embryo.

We describe here the distribution of cellular retinoic acid-binding protein I (CRABP I) in the head of the early mouse embryo from day 8 to day 13 of gestation, using both in situ hybridisation to localise mRNA and immunocytochemistry to localise protein. The distribution of mRNA and protein was found to be identical. CRABP I first appeared in part of the presumptive hindbrain of the presomite embryo and then became localised to rhombomeres 2, 4, 5 and 6. The only other area of expression in the cephalic neuroepithelium was in a part of the midbrain roof. The neural crest and its mesenchymal derivatives, the branchial arches, expressed CRABP I and crest could be seen streaming from the neuroepithelium of individual rhombomeres into particular branchial arches. This suggested a fate map could be constructed describing the rhombomeric origin of branchial arch mesenchyme. Later in development, axons throughout the hindbrain expressed CRABP I. The results are considered in terms of the role of retinoic acid in the specification of neuronal phenotype in the hindbrain and in axon outgrowth.


Immunohistochemical evidence for coexistence of methionine-enkephalin and tyrosine hydroxylase in neurons of the locus coeruleus complex projecting to the spinal cord of the cat.

Previous studies have revealed the presence of pontospinal neurons with either methionine-enkephalin- or tyrosine hydroxylase-like immunoreactivity in the dorsolateral pontine tegmentum of the cat. Using a combined fast blue retrograde transport technique and simultaneous immunofluorescence histochemistry, the present study was designed to reveal the coexistence of enkephalin and tyrosine hydroxylase in cat coerulospinal neurons and to determine if and to what extent the coerulospinal pathway is heterogeneous. Fast blue-labelled neurons with tyrosine hydroxylase- and enkephalin-like immunoreactivities were found in the nucleus locus coeruleus, nucleus subcoeruleus, Kölliker-Fuse nucleus, and the medial and lateral parabrachial nuclei. Approximately 87% of tyrosine hydroxylase-like immunoreactive neurons had enkephalin-like immunoreactivity, whereas about 76% of the enkephalin-like immunoreactive neurons had tyrosine hydroxylase-like immunoreactivity. About 71% of all coerulospinal neurons exhibited both tyrosine hydroxylase- and enkephalin-like immunoreactivities. These findings indicate that coerulospinal activity may lead to spinal cord effects reflecting both norepinephrine and enkephalin activity in most cases but do not rule out each transmitter's isolated functions.


Light microscopic radioautographic localization of somatostatin binding sites in the brainstem of the rat.

The distribution of somatostatin binding sites was investigated by light microscopic radioautography in the brainstem of the rat following in vitro labeling with 125I-Tyr0-DTrp8-somatostatin14. Moderate to high labeling densities were detected within the superior colliculus, the locus coeruleus and subcoeruleus, the parabrachial complex, the nucleus of the solitary tract and the dorsal motor nucleus of the vagus. Most of the white matter was devoid of specific somatostatin binding except for fibers of the glossopharyngeal nerve and the spinal trigeminal tract. In most of the labeled areas, 125I-somatostatin binding was evenly distributed between neuropil and perikarya. In a few instances, however, the binding clearly predominated over nerve cell bodies: namely in the dorsal motor nucleus of the vagus and in the pontine and medullary tegmentum. In the latter two regions, the labeled neurons were identified in adjacent sections by tyrosine hydroxylase immunohistochemistry as belonging to the A5 and A1 catecholamine cell groups, respectively. These findings, together with the confirmed association of somatostatin binding sites with noradrenergic neurons in the locus coeruleus, suggest that interactions with catecholaminergic systems may represent a major mode of action for somatostatin in the brainstem.


Cell type dependent transcription regulation by chick homeodomain proteins.

Five chick homeodomain proteins (CHOXs), CHOX-1.7, -1.1, -1.4, -4.2 and -2.6, had different transcription-regulating activities in a chick cultured cell line, LMH. In particular, CHOX-1.7 highly activated transcription when NP6 was used as the target site whereas CHOX-1.4 did not. This was mainly due to differences in the activation domains since both proteins bound to NP with almost the same affinities in vitro. In LMH cells, they competitively acted on target gene transcription. Moreover, the strength of the CHOX-1.4 activation domain depended on the cell type. These findings suggest that the effect on a target gene is determined by a combination of CHOXs and cell types.


Co-expression of tyrosine hydroxylase messenger RNA 1 and 2 in human ventral mesencephalon revealed by digoxigenin- and biotin-labelled oligodeoxyribonucleotides.

In situ hybridization experiments, using oligodeoxyribonucleotides specific for the two major expressed human tyrosine hydroxylase mRNAs, were performed on human brain sections at the level of the mesencephalon. The specificity of the probes was ascertained by Northern blot experiments carried out with independently in vitro synthesized human tyrosine hydroxylase mRNAs. For in situ hybridization experiments, oligodeoxyribonucleotides were labelled with nucleotides tagged with digoxigenin or biotin molecules. The hybridized oligonucleotides were detected by antibodies coupled with peroxidase and alkaline phosphatase enzymes, which yield, with appropriate substrates, brown and purple products, respectively. The simultaneous detection of the two mRNAs with digoxigeninated and biotinylated probes revealed that these two mRNAs are co-expressed in single cells. The purple product obtained with alkaline phosphatase exhibits a discrete distribution within the dopaminergic cells suggesting these mRNAs are associated with sub-cellular structures. Finally, a heterogeneity in the intensity of the labelling of reactive cells with both probes was visualized as well as the expression of the two mRNA species in neurites.