Induction of c-fos immunostaining in the rat brain after the systemic administration of nicotine. To search for evidence of altered neuronal gene expression in response to exposure to the highly addictive drug nicotine, rat brains were examined by immunocytochemistry for the fos protein after the systemic administration of nicotine. The drug was administered as an IV infusion over 1 h At a dose of 2 mg/kg, the most dramatic nicotine-induced fos nuclear immunostaining was seen in central visual pathways, including the superficial superior colliculus and the medial terminal nu. of the accessory optic tract, in the interpeduncular nu. Notably, many regions with high levels of nicotine binding sites, including the medial habenula, thalamus, substantia nigra, and ventral tegmental area, failed to express the c-fos gene with this schedule of nicotine administration. A minimal increase in fos immunostaining was seen after a nicotine dose of 0.5 mg/kg, with a much greater response after 1 or 2 mg/kg. The response was seen as soon as 60 min after the beginning of the infusion, was maximal at 2-3 h, and declined thereafter. c-fos expression was substantially attenuated in the superficial gray layer of superior colliculus, medial terminal nucleus of the accessory optic tract, and the interpeduncular nucleus by pretreatment with the centrally acting nicotine antagonist mecamylamine, 5 mg/kg IP, but not with the peripherally acting antagonist hexamethonium, 4 mg/kg IP. These observations identify a subset of central nervous system neurons that respond to nicotine with altered expression of the immediate early gene c-fos. These neurons presumably undergo long-term changes in gene expression as a result of acute exposure to high doses of nicotine.


Ethanol alters monoamines in specific mouse brain regions. Ethanol (3.5 g/kg 60 min post-IP injection) produced the following changes in regional brain monoamine levels and in the respective metabolite/neurotransmitter ratios: for the noradrenergic system, MHPG was decreased in the amygdala and increased in the hypothalamus, while the MHPG/NE ratio was increased in the prefrontal cortex and the hypothalamus. For the dopaminergic system, DA was decreased in the olfactory tubercle, DOPAC was increased in the prefrontal cortex and septum, and DOPAC/DA was increased in the prefrontal cortex, septum, striatum, and hypothalamus. HVA was increased in the prefrontal cortex and septum, while HVA/DA was increased in the same regions plus the olfactory bulb. 3MT was decreased in the olfactory tubercle and striatum. The serotonergic system was not altered. The results demonstrate that ETOH produces selective regional changes in the concentration and utilization of monoamines in mouse brain with a predominant influence on dopaminergic systems and a lesser effect on noradrenergic activity.


Eight-arm maze performance, neophobia, and hippocampal cholinergic alterations after prenatal oxazepam in mice. Outbred CD-1 mice were exposed to oxazepam (15 mg/kg PO twice/day) on days 12-16 of fetal life, i.e., at a critical ontogenetic stage of Type II benzodiazepine (BDZ) receptor increase, and fostered at birth to untreated dams. At adulthood, radial arm maze performance, activity-habituation test in an open-field arena (either single 15-min test or three 5-min sessions at 24-h intervals), approach to a novel stimulus object, and amphetamine or scopolamine effects thereon were assessed in male progeny. Overall, the oxazepam exposed (OX) mice were much less efficient in the radial arm maze task than the vehicle exposed (VEH) animals. Pre-test scopolamine injection, but not amphetamine, significantly impaired the arm maze performance of OX mice when compared with the corresponding VEH-scopolamine animals. In separate nonlearned behavioral tasks, prenatal oxazepam did not affect either baseline activity levels in the open field or the response to the amphetamine and the scopolamine challenge, while it considerably increased the latency of first approach to a novel object and produced a deficit of habituation in the course of the subsequent exploratory period. Concomitant investigation at the neurochemical level showed that the adult OX animals had a significant increase in both Bmax and in the affinity (Kd) of cholinergic muscarinic receptors in the hippocampal formation when compared to the vehicle-exposed controls.


Descending pain inhibitory system involved in acupuncture analgesia. The descending pain inhibitory system (DPIS) associated with acupuncture analgesia (AA), caused by low frequency stimulation of an acupuncture point, was identified by the results of lesion and stimulation procedures previously determined to differentiate the afferent and efferent paths in rats. The DPIS starts in the posterior arcuate nucleus and descends to the hypothalamic ventromedian nucleus (HVM) from whence it divides into two pathways: one path, the serotonin mediated path, descends through the ventral periaqueductal central gray (V-PAG) and then to the raphe magnus (RM). The other, the noradrenaline mediated path, descends through the reticuloparagigantocellular nucleus (NRPG) and part of the reticulogigantocellular nucleus (NRGC). The afferent and efferent paths are both present in the RM and NRGC, and were separately identified by means of the analgesia (SPA) produced by stimulation of the separate regions in AA responders and nonresponders, because SPA of these regions in nonresponders produced only efferent pathway mediated analgesia.


Fate of septohippocampal neurons following intracerebroventricular injections of colchicine. Injection of colchicine into the lateral ventricles (ICV) of rats results in a selective loss of neurons immunoreactive for choline acetyltransferase (ChAT) in the medial septum (MS) and a concomitant loss of acetylcholinesterase (AChE) positive fibers in the hippocampus. To determine if this loss of cholinergic cells is due to neuronal death, septohippocampal neurons were retrogradely labeled with fluoro-gold (FG) 1 week prior to the injection of colchicine. Numbers and sizes of FG-labeled and ChAT-immunoreactive neurons were assessed 3, 6, and 10 weeks after ICV colchicine. In line with previous observations, numbers of ChAT-immunoreactive cells were reduced to fewer than 50% of control in the MS and to fewer than 60% of control in the vertical limb of the diagonal band (vDB). Three weeks after ICV colchicine, numbers of FG-labeled neurons were reduced to 48% in the MS and 24% in the vDB. By 6 weeks, the number in the MS decreased further to 31% of control, whereas the number remained at 24% in the vDB. Ten weeks after colchicine, the numbers of retrogradely labeled cells in both the MS and vDB had decreased to 11% of control. The cells which remained were not reduced in cross-sectional area or in diameter. These data suggest that the selective loss of cholinergic neurons in the MS which occurs following ICV colchicine is due to neuronal death and not just loss of ChAT expression.


Effects of medial septal lesions on an operant delayed go/no-go discrimination in rats. Male hooded rats received either a medial septal lesion or a control operation. The rats were then tested on an operant go/no-go discrimination task, first without a delay contingency (20 days), followed by 25 days under an 8-s delay. A discrete trial procedure with symmetrical reinforcement was used. Each trial began with the operant chamber darkened, except for back illumination of a centrally located press panel. Depression of this panel extinguished the back light and initiated a random presentation of either the go stimulus (2800 Hz tone) or the no-go stimulus (10 Hz pulsing light) for 3 s. Stimulus termination initiated the appropriate delay contingency (0 s or 8 s), which was followed by the cued availability of a lever for 2 s. If the rat pressed the lever on go trials, or refrained from pressing the lever on no-go trials, a food pellet was delivered. The data (% correct responses) were averaged over 5-day blocks. Overall, the septal-lesioned rats performed significantly better than the control rats under both O-s delay (p < 0.025) and 8-s delay (p < 0.001) contingencies. Although there was not a significant difference between the groups during the last block of the O-s delay (septals 93% correct, controls 89% correct, p > 0.05) or the first block of the 8-s delay (septals 54% correct, controls 53% correct, p > 0.05), the septal-lesioned animals performed significantly better than the controls during all the remaining phases of the experiment.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of hyperthermia on the effectiveness of MK-801 treatment in the gerbil hippocampus following transient forebrain ischemia. The effects of dizocilipine maleate (MK-801), a noncompetitive N-methyl-D-aspartate (NMDA) receptor/channel antagonist, were tested on the dysfunction of neurotransmitter and signal transduction systems and morphological damage 7 days after transient forebrain ischemia in gerbils. Neurotransmitter system (adenosine A1, muscarinic cholinergic receptor) and signal transduction system (inositol 1,4,5-trisphosphate receptor: IP3, protein kinase C: PKC, L-type calcium channels) binding sites were mapped by in vitro quantitative receptor autoradiography. All ligands used in the present study decreased significantly in the CA1 subfield 7 days after ischemia. In normothermic animals, pretreatment with MK-801 failed to protect against decreased receptor binding in the hippocampus 7 days after ischemia. Moreover, in a morphological study, pre- and posttreatment of MK-801 failed to show protective effects against ischemic neuronal damage. On the other hand, pretreatment of MK-801, without maintaining body temperature, prevented the neuronal death of CA1 subfield 7 days after ischemia. These results weaken the hypothesis that NMDA receptor/channel may play a pivotal role in the pathogenesis of neuronal damage after transient forebrain ischemia.


A detergent-solubilized nicotinic acetylcholine receptor of Caenorhabditis elegans. We have used a spin column assay to study the detergent-solubilized levamisole receptor, a nicotinic acetylcholine receptor of the nematode Caenorhabditis elegans. The receptor can be successfully solubilized in detergent solutions of Triton X-100, Lubrol PX, or sodium cholate. Centrifugal gel filtration assay using the tritiated ligand [3H]meta-aminolevamisole ([3H]MAL) provides a greater signal and a better signal-to-noise ratio for soluble levamisole receptor binding than either polyethylene glycol precipitation or DEAE filter assay with the same ligand. As for membrane-bound receptor, the detergent-solubilized levamisole receptor consists of more than one affinity state. Detergent solubilization appears to increase the affinity of all states for [3H]MAL (Kd for the highest affinity solubilized [3H]MAL binding state, 41 +/- 5 pM). Data is presented on the equilibrium binding and the association and dissociation reaction rates of the receptor. The similar relative efficacy with which various compounds inhibit specific [3H]MAL binding and deficiencies in solubilizable high affinity specific [3H]MAL binding in two receptor mutants show that the solubilized receptor is the same nicotinic acetylcholine receptor that is detected by assaying membrane-bound specific [3H]MAL binding. The detergent-solubilized levamisole receptor is stable at 0 degree to 4 degrees C, making receptor purification feasible.


Evidence for opiate-dopamine cross-sensitization in nucleus accumbens: studies of conditioned reward. We investigated opiate-amphetamine interactions within the nucleus accumbens in responding for conditioned reward. Separate groups of animals received 4-day intra-accumbens treatment with either saline, morphine (0.5 microgram/0.5 microliter), [D-Ala2 NMe-Phe4 Gly-ol5]-Enkephalin (DAMGO; 1.0 micrograms/0.5 microliter), or [D-Pen2,5]-Enkephalin (DPEN; 2.0 micrograms/0.5 microliter). One two subsequent test days, these rats were given a challenge of d-amphetamine (2.0 and 10.0 micrograms/0.5 microliter) and responding for conditioned reward was measured. In the conditioned reinforcement (CR) procedure, food-deprived animals were trained in an initial phase to associate a food reward (primary reinforcement) with a compound stimulus (light/click). In the next phase, a lever was introduced and responding on the lever produced the compound stimulus alone (secondary reinforcement). Previous evidence shows that psychostimulants but not opiates markedly potentiate responding for conditioned reward. In the present design, animals previously treated with either morphine or DAMGO (preferential mu agonists) showed potentiated lever responding following amphetamine challenges, relative to either DPEN- or saline-treated animals. These findings show that prior exposure of nucleus accumbens neurons to mu-selective opiates induces sensitization to the effects of amphetamine. The results are discussed in terms of opioid effects on dopamine transmission and second messenger systems.


Distribution of corticotropin-releasing hormone immunoreactivity in golden hamster brain. The distribution of corticotropin-releasing hormone-immunoreactive (CRH-IR) neurons and fibers was observed in golden hamsters. CRH-IR neurons and fibers were observed within the hypothalamus, thalamus, amygdala, cortex, midbrain, and hindbrain. The largest numbers of CRH-IR neurons were seen within the magno- and parvocellular divisions of the paraventricular nucleus of the hypothalamus and within the septum, bed nucleus of the stria terminalis, preoptic area continuum. The highest density of immunoreactive fibers was observed in the external zone of the median eminence. In addition, many immunoreactive fibers were observed within the bed nucleus of the stria terminalis and the preoptic area. The distribution obtained in hamsters was compared with previously reported distributions from rats, and both were generally similar.


Hippocampal neuronal damage after transient forebrain ischemia in monkeys. To investigate cerebral injury in the monkey due to transient ischemia, monkeys were each subjected to temporary occlusion of eight (bilateral common carotid, internal and external carotid, and vertebral) major arteries. After 0 (control), 5, 10, 13, 15, and 18 min occlusion, blood flow was restored. The monkeys were sacrificed by perfusion fixation 5 days after the operation, and all brain regions were then histologically examined for ischemic neuronal changes induced by the occlusion. The amplitude of EEG signals from skull and scalp became almost isoelectric within 1-6 min after the onset of occlusion. The EEG signals from the hippocampus were markedly attenuated within 1-4 min, although they did not become completely isoelectric. Blood pressure was significantly increased after 10-min ischemia. Five-min occlusion produced no ischemic neuronal changes except a slight increment of glial cells in the striatum and III, V, and VI layers of the neocortices. After 10- to 15-min occlusion, there were ischemic cell changes restricted exclusively to the CA1 subfield of the hippocampus. Eighteen-min occlusion produced more prominent ischemic neuronal damage in the CA1 subfield of the hippocampus, but ischemic neuronal damage was no longer confined to the hippocampus. These results suggest that only the CA1 subfield of the monkey hippocampus could be damaged by mild ischemic insult. We demonstrate that the limited lesion of the hippocampus, especially the CA1 subfield, after 10- to 15-min occlusion of eight arteries in the monkey, produces a model equivalent to human amnesia caused by transient ischemic insult.


Vitamin E attenuates the toxic effects of 6-hydroxydopamine on free radical scavenging systems in rat brain. Rats were divided into the following four groups namely (a) sham-operated control, (b) 6-OHDA-treated, (c) sham-operated vitamin E-fed and (d) Vitamin E-fed treated with 6-OHDA. Total glutathione (GSH) and superoxide dismutase (SOD) were measured in brainstem (BS), striatum (ST), hippocampus, frontal cortex, and nucleus accumbens (N. Acc.). GSH and SOD levels were significantly decreased in all regions of 6-OHDA-treated rats compared to controls. Feeding of vitamin E resulted in a significant reduction of GSH in ST and N. Acc. but caused increases in SOD in BS, ST, and N. Acc. Pretreatment of rats with vitamin E caused significant attenuation of the effects of 6-OHDA on GSH and SOD in most of the brain regions. These results show that vitamin E can spare the scavenging systems from the injurious effects of 6-OHDA.


[Contribution of a hematologic cytogenetic laboratory to the diagnosis, classification and prognosis of various hematologic malignancies]. The presence of an haematological cytogenetic laboratory into a clinical unit allowed to realize a close collaboration between clinicians and cytogeneticists, and to achieve a maximal exploitation of the results of the bone marrow cytogenetic studies in some haematological malignancies. It made it possible to perform sequential bone marrow karyotypes studies during the different phases of a disease: We could thus establish: The diagnostic value of cytogenetic findings for chronic myelocytic leukaemia, secondary acute leukaemia, but not for dysglobulinemias. The prognostic value of cytogenetic abnormalities in chronic myelocytic leukaemia where additional abnormalities to the Ph1 are a hallmark of blastic transformation; in primitive dysmyelopoiesis where they represent a bad prognostic factor; in dysglobulinemias, where they are a signal of terminal evolution. In all these diseases, only complex and multiple abnormalities have a prognostic significance pointing out the emergence of a malignant clone, where as rare and single abnormalities are of no significance. Nowadays, haematological cytogenetic must be bound to molecular biology.


[Iatrogenic disease observed in a hospital setting. II. Analysis of causes and suggestions for novel preventive measures]. In a previous article, we reported the results of a survey of iatrogenic illness in teaching hospitals. In the second part of this study, we shall consider the origins of the 109 adverse events which occurred in a 24-hour period among 1733 hospital patients in a total of 43 units (mean prevalence, 6.28%). In a total of 89 cases (83 of which were drug-related and 6 of which were of various non-instrumental causes), self-treatment and/or poor patient compliance appeared to be the main cause in 16 (18%); negligence or erroneous prescription (non-respect of a contraindication, wrong indication or dosage, excessive duration of treatment, inadequate monitoring, etc...) appeared to be responsible in 27 cases (30.3%). It may be that at least some of these cases of iatrogenic illness due to errors or negligence on the part of the patient and/or physician (48.3%) could be avoided in future by better training of health personnel and improved public health education. The human and financial costs of iatrogenic illness show that risk-prevention measures are required; we propose a number of strategies concerning both initial training in medical school and ongoing education in therapeutics, together with ways of improving the behaviour of both physicians and the pharmaceutical industry. The latter suggestions are addressed at the media and the health authorities; some concern legislation, while others are aimed at improving early public health education, an essential element in this setting.


[Bronchial epithelium and asthma].

[Bronchial epithelium and asthma]. Asthma is a type of chronic desquamative bronchitis and, so far, the role of the bronchial epithelium in this disease has not been fully assessed. There are at least 3 ways by which bronchial epithelial cells may modulate the pathophysiology of asthma: 1. They are resident cells, capable of liberating pro-inflammatory mediators, which may be chemotactic for other cells such as neutrophils and eosinophils. 2. An asthmatic attack results in desquamation of the bronchial epithelium which may have multiple consequences. 3. A repair phase must follow the desquamation although the precise mechanisms underlying this phase are incompletely understood. The level of reparation is likely to play an important role in the prognosis of the disease.


[Importance of hypergastrinemia in diagnosis of Biermer's disease in adults]. Achlorhydria that is due to antrum sparing atrophic gastritis is a characteristic finding in pernicious anemia. As a consequence of this achlorhydria serum gastrin level rise. Usually several signs are required for assessing a pernicious anemia: megaloblastic anemia, low serum cobalamin level, positivity of a Schilling test, low level of intrinsic factor in the gastric juice, existence of anti-intrinsic factor antibodies in plasma and/or in gastric juice. In this study the usefulness of the serum gastrin level is assessed. We report here the results in investigating sixteen patients with a pernicious anemia. All had: megaloblastic changes on bone marrow examination, serum cobalamin level lower than 150 pg/ml. For 14 patients the performed Schilling test was positive. For 2, no Schilling test was done, but anti-intrinsic factor antibodies were elevated. Serum gastrin levels were higher than 250 pg/ml (254 to 1770 pg/ml). In our patients with pernicious anemia, as a result of hypo or achlorhydria, serum gastrin increase was higher than in any other cause of hypochlohydria: anti-acid treatment, vagotomy... Measurement of serum gastrin is easy. High levels are simple to explain if found together with a sparing antrum atrophic gastritis and/or a low level of serum cobalamin. Serum gastrin measurement may be a cornerstone to rapid and reliable diagnosis of pernicious anemia.


[Application of multifactorial statistical calculation searching for a correlation between fat soluble vitamins and cancer]. The aim of this work was to combine computer methodology applied to multi-factorial statistical calculations with liquid chromatography results on the analyses of serum levels of retinol, bêta-carotene and tocopherols in healthy subjects and in patients with various cancers associated with certain tumor markers. Our results show that the serum levels of the vitamins studied differed in cancer patients in comparison to healthy controls. These results also confirm published data showing that serum variations are generally not characteristic of a specific tumor localization, without indicating if these variations are a cause or an effect of tumor development. Computerized multifactorial analysis taking into account the levels of all vitamins and tumor markers chosen, however, enabled us to classify the different tumors in five groups: cancers of the nervous system, urogenital tract cancers, laryngo-pulmonary cancers and two groups of digestive system cancers. This new approach may lead to the constitution of a data bank on the localization of tumors, as well as a reference system for all analyses of serum levels of vitamins associated with tumor markers.


DIROM: an experimental design interactive system for directed mutagenesis and nucleic acids engineering. A computer system DIROM for oligonucleotide-directed mutagenesis and artificial gene design has been designed for better experimental planning and control. DIROM permits searching for optimal oligonucleotides with respect to certain important parameters, namely sufficient energy of oligonucleotide-target hybridization, the secondary structure of oligonucleotide and target DNA, the presence of alternate binding sites in the target DNA and terminal G/C pairs. It can also be used to plan polymerase chain reaction experiments, for optimal primer selection, in sequencing, etc. DIROM enables one to search for both existing and potential restriction sites, to perform vector + target sequence construction. The system consists of a set of original algorithms that formalize the empirical knowledge of oligonucleotide action as primers.


First and second moment of counts of words in random texts generated by Markov chains. An exact expression for the variance of random frequency that a given word has in text generated by a Markov chain is presented. The result is applied to periodic Markov chains, which describe the protein-coding DNA sequences better than simple Markov chains. A new solution to the problem of word overlap is proposed. It was found that the expected frequency and overlapping properties determine most of the variance. The expectation and variance of counts for triplets are compared with experimental counts in Escherichia coli coding sequences.


METASIM: object-oriented modelling of cell regulation. Enzymatic processes and substances are modelled as distinct objects, belonging to a limited number of classes. A set of class definitions in C++ is presented that constitutes an object-oriented programming platform. The latter supports 'biological' data types and functions and facilitates simulation of metabolic and regulatory pathways in living cells. To compute the time-evolution, Euler or Runge-Kutta methods are used, though the latter method compromises a strict object-oriented philosophy. As an example, histone gene expression during embryogenesis of Xenopus laevis is modelled. This object-oriented programming system forms a modelling 'language' which is readily understood by both biochemists and programmers. It allows biological problems to be programmed more easily and correctly and brings the program closer to the biological reality, hence making it more meaningful to bioscientists. Moreover, it can readily be extended to new models by class derivation.


ADSP--a new package for computational sequence analysis. A new protein sequence analysis package, ADSP, is described, of which the SOMAP Screen-Oriented Multiple Alignment Procedure forms an integral part. ADSP (Algorithms and Data Structures for Protein sequence analysis) incorporates facilities to generate potent pattern-recognition discriminators and offers four algorithms with which to scan any NBRF format sequence database: the package has been designed, in particular, to interface with the OWL composite sequence database, one of the largest, distributed non-redundant sources of sequence data of its kind. The system incorporates a powerful method for compound feature analysis, which provides the basis for characterizing and predicting the occurrence of complete protein superfamilies and for pinpointing the emergence of related sub-families. Used iteratively, the approach allows diagnostic performance to be rigorously refined and its efficacy to be assessed both qualitatively and quantitatively, and results in the generation of refined structural or functional features suitable for entry into a database: this compilation of characteristic signatures is distinct from, but complementary to, widely used compendia of pattern templates such as PROSITE.


Clustering cDNA sequences.

Clustering cDNA sequences. A set of programs has been written to quantify the similarities between large numbers of cDNA sequences. This information is used to cluster similar sequences together. The main program can cluster thousands of cDNA sequences per day using a novel, computationally inexpensive algorithm. The clustering information is kept in a small index file so that disk storage requirements are negligible. Using this index file, subsidiary programs create various views and statistical summaries of the entire cDNA sequence collection.


CMAP: contig mapping and analysis package, a relational database for chromosome reconstruction. In the contig mapping and analysis package, CMAP, we provide a foundation for reverse genetics by organizing information about DNA fragments obtained from an organism's genome into a physical map. The user can store information about a particular segment of DNA. This information can be both descriptive, such as any genes contained in a particular DNA fragment, or experimental, such as hybridization profiles or restriction digest patterns for comparison with other fragments. The package can then be instructed to update the physical map or provide information on a DNA fragment within the map, such as its location. The user interface is designed to minimize the learning curve associated with database usage, while eliminating the possibility of entering data outside the ranges of fields through error-checking protocols. Queries are currently accomplished by the use of dynamic SQL (structured query language), which gives the user the ability to build queries based on any combination of the attributes contained within the database without requiring that all possible queries be permanently programmed within the query software. In order to eliminate the need for knowledge of SQL, an interface was designed to allow users to build queries by menu choices. Thus, CMAP is a software package supporting a database for both the production and storage of a physical map as well as being the first step toward the production of a physical mapping workstation.


Limitations of Augustinsson plots.

Limitations of Augustinsson plots. The Augustinsson plot transforms the Michaelis-Menten equation by graphing velocity, v, as a function of velocity/substrate, v/s. The plot is linear with a negative slope for a single class of saturable sites and curvilinear for multiple classes of sites. Statistically, Augustinsson plots suffer from the limitation that there is no true independent variable. Experimental errors form a Gaussian distribution that rests neither parallel to the v axis nor v/s axis, but rather along a line emanating from the origin. Widening the range of s values produces increasingly large errors in the calculated Km and Vm values, especially when multiple sites are present. Exact data containing no error produces accurate constants using either the Michaelis-Menten or Augustinsson equation. With Km and Vm constants previously derived by fitting data with the statistically valid Michaelis-Menten equation, the Augustinsson plot may be used accurately to display multiple site data.


Aligning two sequences within a specified diagonal band. We describe an algorithm for aligning two sequences within a diagonal band that requires only O(NW) computation time and O(N) space, where N is the length of the shorter of the two sequences and W is the width of the band. The basic algorithm can be used to calculate either local or global alignment scores. Local alignments are produced by finding the beginning and end of a best local alignment in the band, and then applying the global alignment algorithm between those points. This algorithm has been incorporated into the FASTA program package, where it has decreased the amount of memory required to calculate local alignments from O(NW) to O(N) and decreased the time required to calculate optimized scores for every sequence in a protein sequence database by 40%. On computers with limited memory, such as the IBM-PC, this improvement both allows longer sequences to be aligned and allows optimization within wider bands, which can include longer gaps.


A finite-difference computer model of solute diffusion in bacterial films with simultaneous metabolism and chemical reaction. This finite-difference computer model is designed to simulate complex diffusion/reaction events in bacterial films. It is modular, each module mirroring closely a particular physical, chemical or biochemical factor. It is capable of handling > 20 diffusing/reacting species, but can be easily expanded or simplified to match particular systems. It was originally designed for modelling the events in dental plaque leading to tooth decay, but should find application in other fields. It allows for ion-exchange interactions with, for example, fixed charges on bacterial surfaces, which can act as pH and cation buffer sites. pH-dependent utilization of substrate is modelled implicitly, combining Michaelis-Menten kinetics with diffusion in a single iterative procedure. Advantages are given for computing diffusion of all other species explicitly using single-species diffusion coefficients, with charge-coupling by means of the algorithm Q-COUPLE. Activity corrections and enzyme pH-dependence are included. Chemical equilibria and mineral deposition/dissolution are computed iteratively node by node. The program is tested against some problems having analytical solutions, and an example is given of its application to demineralization of teeth as a result of bacterial action in dental plaque.


MATCH-BOX: a fundamentally new algorithm for the simultaneous alignment of several protein sequences. Original algorithms for simultaneous alignment of protein sequences are presented, including sequence clustering and within- or between-groups multiple alignment. The way of matching similar regions is fundamentally new. Complete matches are formed by segments more similar than expected by random, according to a given probability limit. Any classic or user-defined score matrix can be used to express the similarity between the residues. The algorithm seeks for complete matches common to all the sequences without performing pairwise alignment and regardless of gap weighting. An automatic screening delineates all the similar regions (boxes) that may be defined for a given maximal shift between the sequences. The shift can be large enough to allow the matching of any region of a sequence with any region of another one. It can also be short and used to refine the alignment around anchor points. The algorithm provides the most likely optimal alignment and a comprehensive list of the alignment dilemma. Duality between automatism and interactivity is provided. Depending on the problem complexity, a final alignment is obtained fully automatically or requires some interactive handling to discriminate alternative pathways.


Dynamic programming algorithms for restriction map comparison. For most sequence comparison problems there is a corresponding map comparison algorithm. While map data may appear to be incompatible with dynamic programming, we show in this paper that the rigor and efficiency of dynamic programming algorithms carry over to the map comparison algorithms. We present algorithms for restriction map comparison that deal with two types of map errors: (i) closely spaced sites for different enzymes can be ordered incorrectly, and (ii) closely spaced sites for the same enzyme can be mapped as a single site. The new algorithms are a natural extension of a previous map comparison model. Dynamic programming algorithms for computing optimal global and local alignments under the new model are described. The new algorithms take about the same order of time as previous map comparison algorithms. Programs implementing some of the new algorithms are used to find similar regions within the Escherichia coli restriction map of Kohara et al.


GCWIND: a microcomputer program for identifying open reading frames according to codon positional G+C content. GCWIND is a microcomputer (IBM-PC compatible) program for the identification of protein-coding open reading frames. The program is similar to the FRAME program, but the latter has only been implemented for a specialized graphics package. The base compositions (%G+C) for each of the three possible reading phases through the DNA sequence are displayed separately, together with the positions of potential translation initiation and termination codons (on the leading and complementary strands), to provide an immediate representation of those regions within the sequence that have coding potential.


Cisplatinum in combination with 5-fluorouracil and citrovorum factor in the treatment of advanced colorectal carcinoma. A phase II trial of citrovorum factor, 500 mg/m2/week, plus 5-fluorouracil, 400 mg/m2/week on day 1, and cisplatin, 20 mg/m2/week on day 2, was carried out in a group of 40 patients with metastatic colorectal carcinoma. A partial response with a mean duration of 8.4+ months was achieved in 24% of patients, a minimal response with a mean duration of 5.4 months was obtained in 6% of patients, and a stabilization of 6.2 months was achieved in 41%. Ten patients (29%) progressed. A 38% partial response rate was seen in patients with advanced rectal carcinoma, whereas no response was obtained in patients with colon cancer. Interestingly, 5 partial responses were seen in 12 patients pretreated with 5-fluorouracil. The overall survival was 9.8+ months. The mean survival of patients who achieved a partial response was 12.0+ months, whereas patients who progressed survived a mean of 6.6+ months. Patients with colon cancer had a mean survival of 8.1+ months, and those with rectal cancer survived a mean of 11.1+ months. This difference was not statistically significant. The treatment was generally very well tolerated, with patients showing mostly grade 1-2 gastrointestinal and/or hematological toxicity.


Immune senescence: mechanisms and clinical implications. There are many changes in the immune response with age, most of which can be related to the involution of the thymus gland and alteration of the distribution and function of T lymphocytes. These changes indirectly influence B-cell function, which not only amplifies the immune deficits, but leads to the increased production of autoantibodies and monoclonal immunoglobulins with age. Thus, immune senescence is characterized by both an immune deficiency and an immune dysregulated state.


Chemical radiosensitizers in cancer therapy. The development of effective low-LET radiation therapy for cancer has been hindered by the lack of consistent differential responses to radiation between tumor and normal tissues. One major difference between many solid tumors and the surrounding normal stroma is the presence of hypoxic foci in solid tumors due to the inadequate supply of nutritional needs as a result of the breakdown of microvasculature. Consequently, failure of conventional radiotherapy and local recurrences are in part attributed to the radioresistant hypoxic cell populations, present in the tumor. Local cure/control rates of a tumor can be increased only by an effective increase in the radiation dose. At the same time, an increase in such a dose would damage the oxic normal stroma, more than the hypoxic tumor cells. Hence, specific modification of tumor radiosensitivity by the use of chemical radiosensitizers, in combination with conventional radiotherapy, is an attractive alternative. Many clinicians and radiotherapists are skeptical about the outcome of using radiosensitizers in patients. Nevertheless, a vast amount of information is currently available regarding the first- and second-generation radiosensitizers both in murine and in human tumors. As a result, it is hoped that eventually a radiosensitizing drug would be discovered/synthesized that will overcome the drawbacks so far encountered in their use in the clinic. In this article, the development of chemical radiosensitizers since the early sixties, the basis for their selection, their mechanism(s) of action, and the results obtained with the various groups of radiosensitizers are reviewed.


Tamoxifen in axillary node--negative breast cancer: multisystem benefits and risks. Adjuvant tamoxifen therapy is associated with modest improvement in disease-free and overall survival in women with invasive axillary node-negative breast cancer. The preponderance of data supporting these general conclusions are from trials in postmenopausal women; in premenopausal women data appear convincing regarding disease-free, but not overall, survival. Firm conclusions regarding magnitude of benefit related to presence of different prognostic factors cannot be drawn at present. In postmenopausal women tamoxifen appears to alter favorably some risk factors for cardiovascular diseases and osteoporosis, which are the most common causes of mortality or morbidity in older American women. Adjuvant tamoxifen is associated with a significantly reduced risk of second primary breast cancer. Major serious risks of tamoxifen therapy include depression, and possibly thrombophlebitis and uterine endometrial cancer. Symptomatic vasomotor and gynecological side effects are frequent. Decision making with women should include assessment of these multisystem benefits and risks.


[Transduction of the visual signal in retinal cells]. Transduction of a visual signal is a complex process. It involves photochemical, enzymatic and ionic reactions. An electrophysiological response is generated on absorption of a photon by a photoreceptor cell's pigment molecule, then propagates to the synapses. The first photochemical reaction, isomerization of retinal, in vertebrates occurs in the photoreceptor cells--rods and cones--of the retina, so changes conformation and activity of a pigment-bound protein, rhodopsin, in membranes of intracellular discs. Rhodopsin becomes enzymatically active and catalyses the activation by GTP of a great number of transducins, which in turn activate cGMP phosphodiesterase. This enzymatic chain propagates and greatly enhances hydrolysis of cytoplasmic cGMP. One photon incites hydrolysis of 10(5) cGMP molecules in 100 ms. Local cGMP decrease frees it from specific binding sites in cytoplasm occupied by proteinaceous canals in cell membrane around the activated disc. In darkness high cGMP concentration, hence binding, kept canals open, maintaining high cellular cation permeability, especially to Na+, and a strong cellular depolarization. Ca2+ influx, also allowed, balanced Na+ movement. Canal closure induces local hyperpolarization, the first electrophysiological response, which propagates through the cell to synaptic contacts. It also lowers intracellular Ca2+ concentration which initiates cGMP synthesis--from GTP by a guanylate cyclase controlled negatively by recoverin, a calcium-dependent protein--to restore cGMP towards at rest level. Although all macromolecules involved in this now fairly complete scheme have been isolated and characterized, cloned and sequenced, no three-dimensional structure has yet been established. The proteins are membrane-bound rather than in independent crystal form, which renders such structural studies difficult.(ABSTRACT TRUNCATED AT 250 WORDS)


[Activity of polar glycopeptidolipids from Mycobacterium chelonae in the reversal of chemically induced leukopenia in mice]. Intraperitoneal administration of polar glycopeptidolipids extracted from Mycobacterium chelonae (GPLp-Mc) has led to reversal of Doxorubicin-induced leucopenia in a manner comparable to that effected by GM-CSF administered in a dose of 100 IU (2.5 micrograms/kg). The mode of action and the toxicity of this product are being studied. Results obtained on the mouse indicate that it would be worthwhile to undertake tests in man aimed at studying the effect of GPLp-Mc on chemotherapy- and radiotherapy-induced leukopenias, once toxicological studies have been carried out.


[Metabolic effects of phosphate receptors stimulating the ileal transport of calcium]. Several compounds, either metabolizable such as sorbitol and creatine or unmetabolizable such as L-xylose or ethyl-ethanolamine, increase ileal calcium transport, similarly to the well documented effect of lactose. These compounds increase the duration time but not the rate of calcium transport. They have no specific effect on intestinal calcium absorption but they prolong the presence of soluble calcium in the ileal loop, thus allowing the calcium absorption. This stimulating effect of these compounds on mineral absorption is associated with an increase of ileal mucosal ATP content.


[Description of a tissue culture technique using tissue from mussel (Mytilus edulis) for the preparation of chromosomes]. Explants from mantle and foot tissues of adult mussel were grown in culture tubes containing a medium composed of Eagle's Basal Medium supplemented with salts, Hepes buffer, egg yolk and antibiotics. The cultures were maintained at 18 degrees C and pH 7.50, without medium renewal. After 6-7 days, the cultures were stopped and harvested for slide preparation. Numerous metaphase spreads that were good enough for karyotyping were consistently obtained. This method may prove to be a reliable source of actively dividing cells that is a prerequisite for extensive chromosome structure analyses in the bivalves.


[Mollicute Acholeplasma florum possesses a gene of phosphoenolpyruvate sugar phosphotransferase system and it uses UGA as tryptophan codon]. Acholeplasmas are mollicutes which do not require sterol for growth and do not possess a phosphoenolpyruvate-dependent sugar-phosphotransferase system. In contrast to spiroplasmas, mycoplasmas and ureaplasmas, they utilize UGA as a stop codon and not as a tryptophan codon. Acholeplasma florum, because of its metabolic properties and its close phylogenetic relationship to members of the genera Spiroplasma and Mycoplasma, does not seem to be an acholeplasma senso stricto. Here, we present molecular data to support this hypothesis. We have detected a gene coding for one of the components of the phosphotransferase system (PTS) in A. florum. In addition we demonstrate that the organism uses UGA as a tryptophan codon and not as a stop codon. These findings offer strong evidence, along with an earlier phylogenetic proposal, that A. florum is not a member of the genus Acholeplasma.


[Spatial memory of Meriones libycus (Rodienta, Gerbillidae)]. Meriones libycus, a desert rodent with a large home range, has a good spatial memory over at least 90 days, which makes it able to travel for a long way and come back to its nest. Males have a better spatial memory than females.


An enzymatic determination of hydrogen peroxide by using spectrophotometry. A new spectrophotometric method for determining low hydrogen peroxide concentrations by using horseradish peroxidase in the presence of NADH at pH 7.5 has been described. Both total NADH consumption and initial reaction rate may be used for the determination. Using the NADH consumption, a linear response with respect to hydrogen peroxide was observed in the concentration range 7 x 10(-8)-2.5 x 10(-6) M. Due to the presence of superoxide dismutase, hydrogen peroxide is partly regenerated and an amplification of the signal results, which explains the sensitivity.


[Respective role of mifepristone (RU486) binding to blood and tissue proteins in its quantitative distribution in the body]. In plasma, mifepristone (RU486)-alpha 1-glycoprotein acid (AGP) interaction follows a quickly saturable process within the range of therapeutic concentrations. The high affinity to AGP, Kap = 8 x 10(6) M-1, suggests that the tissue distribution of the drug is possibly impaired. Checking this hypothesis was done by simulating its body distribution between plasma proteins and tissues. Two methods were used. The first was to calculate the number of available binding sites in both plasma (Np) and tissues (NT), to measure the relevant association constants Kap and KaT, and to consider that mifepristone partitioned according to their ratios, expressed as [NpKap]/[NpKap + NTKaT] for the plasma and [NTKaT]/ NpKap + NTKaT] for the tissues. The second method used equations relating the apparent volume of distribution and the plasma unbound fraction of mifepristone with volumes of physiologic spaces to calculate the percentage of drug located in plasma, extracellular and intracellular fluids. The results yielded by the two methods are close. They show that with AGP levels within a normal range (18.4 microM), only 18% of mifepristone is bound to AGP and the remaining part 82% is located in tissues. By contrast, when AGP level dramatically increases (300 microM) most of the dose (77%) is retained in plasma. These results suggested that when AGP concentration increases, the efficacy of therapeutic concentration of mifepristone may be partially decreased if only the unbound fraction of this synthetic hormone were biologically active.


Inhibition of sulfur mustard-increased protease activity by niacinamide, N-acetyl-L-cysteine or dexamethasone. The pathologic mechanisms underlying sulfur mustard-induced skin vesication remain undefined. Papirmeister et al. (1985) have postulated a biochemical mechanism for sulfur mustard-induced cutaneous injury involving DNA alkylation, metabolic disruption, and enhanced proteolytic activity. We have previously utilized a chromogenic peptide substrate assay to establish that human peripheral blood lymphocytes exposed to sulfur mustard exhibited enhanced proteolytic activity. In this study, compounds known to alter the biochemical events associated with sulfur mustard exposure or to reduce protease activity were tested for their ability to block the sulfur mustard-increased proteolysis. Treatment of cells with niacinamide, N-acetyl-L-cysteine, or dexamethasone resulted in a decrease of sulfur mustard-increased protease activity. Complete inhibition of sulfur mustard-increased proteolysis was achieved by using protease inhibitors (antipain, leupeptin, and 4-(2-aminoethyl)-benzenesulfonylfluoride). These data suggest that therapeutic intervention in the biochemical pathways that culminate in protease activation or direct inhibition of proteolysis might serve as an approach to the treatment of sulfur mustard-induced pathology.


[Famotidine does not modify the plasma levels of lidocaine for spinal anesthesia. A comparative study of famotidine and cimetidine]. H2 receptor antagonists are often used to reduce gastric acidity prior to general or regional anaesthesias. However cimetidine and ranitidine have been found to increase plasma levels of local anaesthetics such as lidocaine. This study aimed to compare famotidine--another H2 receptor antagonist--with cimetidine in this respect. 33 male patients 70 years old or more, scheduled for surgery under spinal anaesthesia and randomized into 3 groups, received either diazepam 0.15 mg.kg-1 and famotidine 20 mg (group A) or diazepam and cimetidine 200 mg (group B) or diazepam only (group C) on the previous night and 90 minutes before spinal anaesthesia by hyperbaric lidocaine 5%, 3 ml. Arterial blood samples were taken 3 minutes after spinal injection then every 15 minutes unto 90 minutes after the first sampling. Lidocaine plasma levels were measured by fluorescence polarization immunoassay and the results were statistically analyzed. In famotidine group lidocaine levels were intermediate between B and C groups levels, and the increase was of lesser duration than in B group. Elimination mechanism of lidocaine with and without H2 inhibitor is briefly discussed. Therefore famotidine would appear to be more convenient than other H2 receptor inhibitors whenever antacid protective effects are sought after prior to regional anaesthesia.


[A comparative study of the cost of open-circuit as opposed to closed-circuit ventilation]. The authors compared two open randomized groups of patients undergoing surgery through general anaesthesia. Group 1 consisted of 54 patients ventilated by a Siemens 900 B ventilator in open circuit, and group 2, 56 patients ventilated by an ELSA de Gambro ventilator in a closed circuit. Comparative hour cost for nitrous oxide (N2O), oxygen (O2) and halogen gas, Enflurane, Isoflurane, was noted. All patients received the same regimen of anaesthesia and the two groups were identical in age, weight, surgery, respiratory volume and ventilation time. The evaluation of comparative hour cost included specific materials of close circuit ventilator: CO2 filter (Aridus), Lime. Were excluded maintenance and gas consumption expenditures before patient connected to the ventilator. The total hour cost (O2, N2O, specific materials for close circuit, without halogen gas) was 8.23 FF in closed circuit against 13.28 FF in open circuit, an economy of 38.27%. Hence, for oxygen, the hour cost was 0.70 FF in open circuit against 0.27 FF in closed circuit (gain of 65.3%). For nitrous oxide, the hour cost in open circuit was 12.50 FF against 2.44 FF in closed circuit (80.5%). For Isoflurane, the open circuit hour cost was 41.38 FF against 22.44 FF in closed circuit (47%). For Enflurane, the open circuit hour cost was 14.17 FF against 5.94 FF in closed circuit (58.1%). And, lastly for Enflurane, open circuit hour cost was 14.17 FF against 5.94 FF in close circuit, gain of 58.1%. These "modest" economy against those found in previous studies can be explained by the long-time duration of ventilation, saturating time in open circuit more or less long, depending on the physician, specific materials for closed circuit ventilation--lime, CO2 filter--in not taken into account, the hour cost of O2 + NO2 goes from 8.23 FF to 2.71 FF, and the gain against the close circuit becomes 79.6%: reducing hour cost by 5 times. In order to improve the effective cost of close circuit, the authors proposed: the use of closed circuit ventilation for more than 3 hours surgery, gas saturation in closed circuit after denitrogenation--which demands the use of halogen infjectors, and lime in containers cheaper than disposable cartridges. Respecting the above criteria, the total hour cost in close circuit fell to 4.90 FF, gain of 63% against open circuit. For O2 et N2O, the hour cost goes from 1.34 FF in close circuit to 13.28 FF in open circuit, 90% economy.(ABSTRACT TRUNCATED AT 250 WORDS)


Expression of heat shock-regulated human growth hormone genes containing or lacking introns by NIH-3T3 and Wish cell lines. A plasmid containing the complete genomic DNA of the human growth hormone (ghGH) comprising four introns and driven by the human promoter of the human gene of the 70 kDa heat shock protein (hsp70) has been used to transfect mouse NIH-3T3 and human Wish cells. Selected cell lines were characterized for stable hGH secretion. Similarly in the same NIH-3T3 cells, the stable expression of the same plasmid construct, but containing the complementary DNA of the hGH gene (chGH), was compared in terms of the effect of introns on heterologous protein synthesis. Genomic hGH recombined cells synthetized, in a heat regulated fashion, matured hsp70/hGH hybrid mRNA able to drive the secretion of a 22 kDa polypeptide. Like the natural hGH, this polypeptide expressed the functional hormonal activity of prolactin on casein secretion by mammary cells. The time course of hGH secretion was prolonged in ghGH transcripts, while that of mRNA degradation appeared delayed, especially in Wish cells, as compared to chGH expression. In the human Wish cells the decay of endogenous hsp mRNA has been compared to that of recombinant hsp mRNA, demonstrating that this human hsp70/hGH hybrid mRNA was present in the cytoplasm during a longer period than the human endogenous hsp70 mRNA. In conclusion, similar levels of expression and resulting gene products were expressed from the chGH or the ghGH gene in an inducible manner.


Comparison of in vivo acute lethal potency and in vitro cytotoxicity of 48 chemicals. The cytotoxicity of 48 compounds included in the MEIC (Multicenter Evaluation of In Vitro Cytotoxicity) list was determined in cultures of rat hepatocytes, McCoy, and MDBK cells. The average minimum concentration of each compound inducing cytotoxicity was measured in each cell type. The cytotoxicity values were then compared with published oral LD50 values for rats and mice. The logarithmic transformation of in vivo toxic doses and the corresponding in vitro cytotoxic concentrations showed a statistically significant correlation between the in vitro and in vivo values. The results show that an accurate in vivo LD50 dose could be predicted from in vitro data for at least 75% of the selected compounds. It is hoped that this finding will not only stimulate others to pursue in vitro technique but will eventually lead to elimination of the in vivo LD50 test.


[Severely burned patients and atracurium: the train-of-four response]. Severely burned patients show a relative hyposensitivity to atracurium. The neuromuscular block obtained in 20 patients with major thermal injury after 0.75 mg.kg-1 i.v. of atracurium, by muscular activity recording (train of four-TOF), was studied. The results confirmed the resistance to atracurium, which mechanism is unknown. Even the doses administered, 50% higher than clinical dose (0.3-0.5 mg.kg-1), proved insufficient in order to obtain a complete neuromuscular block in severely burned patients.


Toxic effects of sulfur mustard on respiratory epithelial cells in culture. Sulfur mustard (SM) is known to induce cutaneous injury and to cause acute damage to the respiratory tract. Although skin vesication has been demonstrated on human epidermal keratinocytes in culture, no study has been carried out to analyze the effects of SM on the ultrastructural and functional activity of surface respiratory epithelial cells. To evaluate this SM toxicity, we developed an in vitro model of respiratory epithelial cells in primary culture. The study was performed on surface epithelial cells from rabbit trachea cultured according to the explant-outgrowth technique. The functional activity of the cultures was evaluated by measuring the ciliary beating frequency (CBF) of the ciliated cells with a videomicroscopic method. The morphological aspects of the cells were analyzed by light and electron microscopy. Addition of 0.1 mM SM directly into the culture medium produced a sudden and irreversible CBF inhibition, first observed after 2 hr on the ciliated cells of the outgrowth periphery. The arrest of the ciliary beating progressively reached the whole surface of the outgrowth and was simultaneously observed with a detachment of the outgrowth cells. It began at the outgrowth border, leading to the exfoliation of cell sheets, and then to the whole culture after 48 hr. Morphological damage was expressed by intense vacuolisation and disorganization of cytoplasmic and nuclear structures. These findings suggest that the detachment of the respiratory epithelial cells from the matrix represents a major toxic effect of 0.1 mM SM. SM dramatically affects the viability of respiratory epithelial cells in culture. Moreover, the sudden CBF inhibition is more likely due to the death of the ciliated cells than to a specific ciliotoxic effect of SM.


[The effect of the local anesthetic temperature on spinal anesthesia using 0.5% bupivacaine]. The aim of this study was to compare spinal anaesthesia with 0.5% bupivacaine at 20 degrees C and 5 degrees C. One hundred two patients ASA I-II were randomized into two groups. After atropine and IV fluid administration, patients of group I received intrathecally at L3-L4 level 0.02 ml.cm-1 of plain 0.5% bupivacaine at 20 degrees C, while patients of group II received the same solution and the same doses but at 5 degrees C. No significant difference was found between the times for installation of sensory and motor blocks nor between the durations of motor block in the two groups. On the other hand significant differences were found between the durations of sensory block in group I (283.33 min +/- 9.40 min) and group II (314.35 +/- 8.80 min) (P < 0.001) and all the more (P < 10(-9)) between the cephalad block extensions in group I (D 7.45 +/- 0.14) and group II (D 4.82 +/- 0.20). Therefore plain bupivacaine 0.5%, isobaric at 20 degrees C, seems to get the properties of an hyperbaric solution when cooled at 5 degrees C.


[The intravascular transfer of glycine during percutaneous kidney surgery]. Transurethral prostatic resection using a 1.5% glycine solution causes a well known clinical and metabolic syndrome called TURP-syndrome. Recent development of percutaneous renal surgery is responsible of several similar accidents. In a prospective study of 150 patients (mean age: 35 +/- 10) subjected to a percutaneous nephrolithotomy, the natremia and the amino acid content of the plasma were measured preoperatively and immediately postoperatively by chromatography. The study shows that there is a post-nephrolithotomy syndrome in two per cent. This syndrome contains a hemodilution with hyponatremia and reabsorption of irrigation fluid. Glycolemia, serinemia and threoninemia increase significantly. These modifications have a good correlation between them except for the natremia. Variability of results in this study and in the literature is explained by difficulty and duration of surgery, volume of glycol used, increasing intrarenal pressures and sudden opening of vessels peroperatively. The gravity of post-nephrolithotomy syndrome requires to change the irrigate solute and use normal saline solution when it is possible.


[The importance of neurologic monitoring in anesthesiology]. Owing to the recent advances of bioelectric signals computerized treatment a real monitoring of anaesthesia is now possible, either by EEG or by classical or event-related evoked potentials methods. Available methods and their indications and difficulties are discussed with the help of some clinical examples.


[Practical aspects of the laryngeal mask]. The laryngeal mask (LM) is increasingly used. An overview is made: description, insertion, cardiovascular responses, advantages, LM and aspiration, LM as an aid to ventilation, indications, LM in difficult intubations, contra-indications, sterilization and cost.


Paracetamol anaphylaxis.

Paracetamol anaphylaxis. We describe four cases of anaphylaxis to paracetamol without co-existing aspirin intolerance and review the Australian experience in paracetamol anaphylaxis. Paracetamol sensitivity may differ in the mechanism from sensitivity to aspirin and other non-steroidal anti-inflammatory drugs as previously believed.


Lability in complement receptor mobilization of granulocytes in patients with bronchial hyperreactivity. The recruitment of activated granulocytes to bronchial mucosa seems to be involved in the prolonged inflammatory response observed in asthma and probably associated with bronchial hyperresponsiveness. We studied the patients with bronchial hyperresponsiveness with respect to the expression of complement receptor type 1 (CR1), its variability and readiness to become mobilized on peripheral granulocytes from patients with bronchial hyperresponsiveness. CR1 expression and its hourly variation was significantly (P less than 0.02, P less than 0.01 respectively) higher in the patient group compared with the control group. In addition the ability to mobilize CR1 spontaneously at +37 degrees C correlated to the variation of CR1 expression that occurred during a 4 hr period. These findings indicate that granulocytes from patients with bronchial hyperresponsiveness have a higher degree of variation in CR1 expression that correlates to their ability to mobilize CR1 and may reflect a more pronounced lability.


Longitudinal variability of skin prick test results. The skin prick test (SPT) is a commonly used procedure for assessing a specific sensitization. The longitudinal variability of test results is of interest for clinical as well as epidemiological investigations. The sensitization to four common aeroallergens (grass pollen, birch pollen, Dermatophagoides pteronyssinus, cat dander) is investigated within the framework of three consecutive SPTs at 11-month intervals for a population of 587 schoolchildren. The prevalence of sensitization based on a weal diameter of at least 2 mm was between 12.9% (cat dander) and 23.9% (grass pollen) in the initial testing. The positive predictive values of the initial SPT were between 75.3% (birch pollen) and 88.2% (cat dander) for the two subsequent SPTs. In the case of initially negative tests with positive second and third SPTs the incidence ranged between 3.2% (cat dander) and 4.3% (birch pollen) per year. A clear increase in the intensity of reaction in subsequent tests was observed in a number of probands testing positively in the initial SPT. In conclusion, our data indicate a high long-term stability of a specific sensitization to aeroallergens in SPT.


Urinary excretion of angiotensin I, II, arginine vasopressin and oxytocin in patients with anaphylactoid reactions. Human urine samples, purified on octadecasilyl-silica cartridges, contained immunoreactive angiotensin I, II, arginine vasopressin and oxytocin. The daily excretion of these peptides in healthy volunteers was 190.00 +/- 38.43 (n = 12), 17.48 +/- 3.09 (n = 12), 63.43 +/- 14.84 (n = 8) and 13.52 +/- 1.42 (n = 7) pmol/24 hr, respectively (mean +/- s.e.m.). Patients with a history of anaphylactoid reactions to drugs or food additives showed clinical symptoms such as urticaria, flush, nausea, dizziness and hypotension after oral provocation with cyanocobalamine, propyphenazone, acetylsalicylic acid and sodium benzoate. In five of the seven patients, angiotensin I and II were increased several fold in the urine fractions after symptoms were reported. The average increase in the urine concentration of both peptides was fourfold and 5.5-fold. In three out of five patients, the mean excretion of arginine vasopressin and oxytocin immunoreactive material was also elevated by a factor of 5.7 and 4.4, respectively. Oral provocation with a placebo failed to elicit anaphylactoid symptoms or an increase in the urine levels of angiotensin I or angiotensin II. Angiotensin I and angiotensin II-like immunoreactivity could be characterized on HPLC as Ile5-angiotensin I, Ile5-angiotensin II and angiotensin II metabolites. HPLC characterization of immunoreactive arginine vasopressin and oxytocin in two different gradient systems showed retention times different than the retention times of the corresponding synthetic standard peptides indicating that both peptides are not authentic AVP and OXT. These results suggest that angiotensin I and angiotensin II may be involved in the clinical events observed during some forms of anaphylactoid reactions.


Autologous bacteria induce chemotaxis of peripheral blood mononuclear cells (MNC) from non-atopic asthmatics. The chemotactic response of peripheral blood MNC from healthy subjects and non-atopic asthmatics against the respective pathogen isolated and cultured from sputum of individual patients was investigated. We found that the wide range of concentrations of autologous bacteria induced chemotaxis of MNC from asthmatics but showed no influence on MNC from healthy subjects. This finding might explain the mechanism of lymphocyte accumulation in the lungs of non-atopic asthmatics.


Allergy. Conventional and alternative concepts. A report of the Royal College of Physicians Committee on Clinical Immunology and Allergy. Allergy is an exaggerated response of the immune system to external substances. It plays a role in a wide range of diseases. In some, such as summer hayfever, the symptoms are due entirely to allergy. In other conditions, particularly asthma, eczema and urticaria, allergy plays a part in some patients but not all. In these situations, allergy may have either a major role or provide just one of many triggers. In an individual patient's illness, the importance of allergy may change with time. The most common allergens (substances causing allergy) are grass and tree pollens, the house dust mite, products from pets and other animals, agents encountered in industry, wasp and bee venom, drugs, and certain foods. Food allergy presents a particularly difficult problem. Some individuals who react to food suffer from food allergy in its strict sense but in others there is no evidence of an alteration in the immune system. Here the term 'food intolerance' is preferable. Conventional doctors treat allergy by allergen avoidance--where this is possible--and drugs that relieve symptoms. In a few selected cases, in which other methods have failed, immunotherapy (desensitisation or hyposensitisation) is recommended. Although patients who consult practitioners of alternative allergy may do so by preference, it is often also because they are dissatisfied with the conventional approach to diagnosis and treatment, or because they have conditions which conventional doctors do not accept as having an allergic basis. There is a very wide range of alternative approaches to allergy, including the methods used by clinical ecologists and other treatments such as acupuncture and homoeopathy. Hypnosis may have a small role to play in helping the asthmatic and similar effects have been suggested for acupuncture. Furthermore, it is likely that there are still many active ingredients in medicinal plants used by herbalists but these need to be clearly identified and purified before their usefulness can be evaluated properly. Apart from these situations, we have yet to be convinced by substantial evidence that any of the other alternative methods of diagnosing or treating allergic disease are of proven value. There have, however, been many false and misleading claims and serious harm may be caused by misdiagnosis or delays in appropriate treatment. The public should be warned against costly methods of diagnosis and treatment which have not been validated.(ABSTRACT TRUNCATED AT 400 WORDS)


Secular trends in the incidence of postmenopausal vertebral fractures. Several studies suggest secular increases in hip fracture incidence through this century, but little is known about such trends for vertebral fracture. We have examined changes in the incidence of clinically ascertained vertebral fractures among Rochester, Minnesota residents aged 35-69 years, that were first diagnosed between 1950 and 1989. Our results indicate no overall increase in incidence over the 40-year period. Categorization of fractures according to the level of preceding trauma, however, revealed a significant increase in the incidence of fractures following moderate trauma among women aged 60-69 years. This increase occurred between 1950 and 1964, and leveled off thereafter. Rates for severe trauma fractures among postmenopausal women, and for vertebral fractures from any cause among younger men and women, remained stable. The rise in moderate trauma fractures in postmenopausal women paralleled that for hip fractures in Rochester and began to plateau at around the same time. It might have resulted from increased diagnosis of vertebral fractures, but the increase in hip fracture incidence is inconsistent with this explanation. An increase in the prevalence of osteoporosis, however, might account for the trend in both types of fractures.


Annual injection of vitamin D and fractures of aged bones. In order to investigate the effect of a supplementation of vitamin D in the prophylaxis of fractures of the bones of aged people, an annual intramuscular injection of ergocalciferol (150,000-300,000 IU) was given to two series of aged subjects: first to 199 (45 male) of 479 subjects (110 male) aged more than 85 years who were living in their own home, and second to 142 (29 male) of 320 (58 male) subjects aged 75-84 and living in a home for aged people. This prospective series was divided into treatment groups according to month of birth. These injections were given annually from September to December in the years 1985-1989, two to five times to each participant. The fracture rates, laboratory values, vitamin D levels, possible side effects, and mortality were followed until October 1990. A total of 56 fractures occurred in the 341 vitamin D recipients (16.4%) and 100 in 458 controls (21.8%) (P = 0.034). The fracture rate was about the same in both outpatient and municipal home series. Fractures of the upper limb were fewer in the vitamin D recipients, 10/341 = 2.9% (P = 0.025), than in the controls, 28/458 = 6.1%, during the follow-up. A similar result was obtained in fractures of ribs, 3/341 = 0.9% and 12/458 = 2.6%, respectively. Fractures of the lower limbs occurred almost as frequently, 31/341 = 9.1%, among the vitamin D recipients as among the controls, 49/458 = 10.7%. The fracture rate was higher in females (22.2%) than in males (9.5%). The fractures were fewer in the vitamin D recipients only in females.(ABSTRACT TRUNCATED AT 250 WORDS)


Crystallographic lattice refinement of human bone. X-ray diffraction studies on bone microsamples (human iliac crest of 87 individuals aged 0-90 years) reveal that certain crystallographic parameters such as unit cell volume of bone apatite, and half-width of (002)-reflection are well correlated with age and with type of tissue (corticalis and spongiosa). Similar to inorganic apatite, the lattice parameters of bone apatite are intensely affected by ionic substitutions and vary mainly due to exchange of hydroxyl- and carbonate-apatite and, to a minor extent, of fluor- and chlorapatite.


Antiidiotypic PTH antibodies as a cause of elevated immunoreactive parathyroid hormone in idiopathic hypoparathyroidism, a second case: another manifestation of autoimmune endocrine disease? A 69-year-old man became hypocalcemic under medical observation. The hypocalcemia occurred in the presence of circulating immunoreactive parathyroid hormone (PTH). Common causes of secondary hyperparathyroidism were excluded, as was PTH resistance using PTH infusions. The immunoreactive PTH was examined in detail. PTH immunoreactivity (1) was not retained on a C18 SPE-column, suggesting unusual molecular or physicochemical properties, unlike bona fide PTH; (2) was precipitated with 15% PEG, indicating a molecular size far in excess of native PTH; (3) had an apparent molecular size similar to immunoglobulins on size exclusion chromatography; (4) was retained on affinity chromatography with both Protein A and anti-hIgG antibodies. These data lead us to conclude that the immunoreactive PTH was due to antiidiotypic PTH autoantibodies. No significant quantities of true PTH were found in the patient's serum suggesting that his hypoparathyroidism was a result of PTH deficiency. Autoimmunity might explain the occurrence of both processes if an arrested antiidiotypic antibody cascade is assumed.


Incorporation of sodium fluoride into cortical bone does not impair the mechanical properties of the appendicular skeleton in rats. Clinical studies on the use of sodium fluoride (NaF) in osteoporotic patients have demonstrated increased spinal bone mass without a reduction in vertebral fracture incidence, and a trend towards reduced appendicular bone mass with an increase in peripheral fracture incidence. As previous reports have suggested that NaF becomes incorporated into bone's crystal structure, possibly affecting bone strength, we sought to examine the relationship among bone fluoride content, bone mass, and skeletal fragility. Twenty-one-day-old female Sprague-Dawley rats were treated with four different doses of NaF. The tibiae were subjected to histomorphometric and biochemical analyses, and the femora were tested in torsion for the properties of strength, stiffness, energy storage capacity, and angular deformation. The results showed that over 50% of the skeleton in these rats was turned over in the presence of NaF. The four different doses resulted in a linear increase in bone F concentration and suggested excellent absorption and incorporation of this drug. No changes in histomorphometric indices of bone formation or turnover were found. Despite the large fraction of bone formed during NaF treatment, and the linear increase in bone fluoride content in relation to dose, there were no changes observed in any of the mechanical properties. These results suggest that, even extensive incorporation of fluoride into bone, in the absence of an effect on bone mass or remodeling, does not significantly alter its capacity to withstand mechanical loads.


Cultured embryonic bone shafts show osteogenic responses to mechanical loading. Pairs of 17-day embryonic chick tibiotarsi were removed and maintained in organ culture. One of each pair was subjected to a single 20-minute period of intermittent loading at 0.4 Hz, producing peak longitudinal compressive strains of 650 microstrain (mu epsilon). In the 18-hour culture period following loading, alkaline phosphatase levels in the osteoblasts of the loaded tibiotarsi were maintained whereas in controls they declined. In situ hybridization using a collagen type I cRNA riboprobe showed a substantial increase in expression of mRNA for collagen type I in the periosteal tissue of bones that were cultured for 18 hours after loading compared with that in similarly cultured controls and bones cultured for 4 hours. These results demonstrate that appropriate loading of embryonic chick bones in organ culture elicits adaptive regulation of matrix synthesis as evidenced by increased expression of the gene for type I collagen and alkaline phosphatase activity. This model may be useful as it must contain all the obligatory steps between strain change in the matrix and modified osteogenic activity.


Effect of a five-week swimming program on rat bone: a histomorphometric study. To specify the exercise-induced changes on different skeletal sites, the effect of a 5-week endurance swim training was studied in rats. Eighteen Lyon strain (Sprague-Dawley) 5-week old female rats were divided into nine sedentary and nine swimming rats. Each swim training session was increased by 15 minutes from 2-6 hours per day. A histomorphometric study was performed at the primary and secondary spongiosa of the distal femur and at the secondary spongiosa of lumbar and thoracic vertebral bodies. After training, bone loss was observed in the secondary spongiosa of lumbar vertebral bodies (24.7%) and in the primary spongiosa of distal femur (15.2%). A tendency to bone loss was also detected in the secondary spongiosa of distal femur (10.8%), whereas no change was detected in thoracic vertebral bodies. In secondary spongiosa, bone loss was accompanied with a thinning of trabeculae. Total eroded surfaces and osteoid surfaces were significantly decreased in the three studied skeletal sites, suggesting a decreased bone turnover. The decreased thickness of osteoid seams in both lumbar vertebrae and distal femur could mean that the osteoblastic activity has also been altered at the cell level, leading to thinning of trabeculae. Five-week swim training with such duration and intensity of exercise appears unable to increase bone volume in rats and, therefore, causes adverse effects. The three studied bones seemed to adapt differently to experimental conditions. The lack of ground reaction forces induced by water immersion might have contributed to the observed bone loss. "Normal" gravity would be an important cofactor in the osteogenic effects of exercise.


Competitive adsorption of magnesium and calcium ions onto synthetic and biological apatites. Magnesium (Mg) is a conspicuous constituent of hard tissues but its possible role in biomineralization is poorly understood. It is possible that Mg2+ adsorbed onto bioapatites may contribute to the modulation of crystal growth as such inhibitory activity has been reported for synthetic apatites. The present study was undertaken to determine the adsorption isotherms of Mg ions onto synthetic apatites and biominerals in tooth and bone tissues in the presence of other ions of natural occurrence. Synthetic crystals used as adsorbents were hydroxyapatite and, as a better prototype for the biomineral, Mg-containing carbonatoapatite. Human enamel and dentin materials were obtained from extracted, caries-free, permanent teeth. Porcine dentin materials at two developmental stages were obtained from erupted deciduous and unerupted permanent teeth of a 6-month-old slaughtered piglet. Porcine bone was obtained from the cortical portion of the mandible of the same animal. All biomineral samples were pulverized and then treated by plasma ashing (deproteination) at about 60 degrees C. Each of the powdered samples was equilibrated in solutions containing various initial concentrations of Mg2+, Ca2+, and Na+ (or K+) as nitrate salts. Following equilibration, concentrations (and activities) of magnesium and calcium ions in the experimental solution were determined. The pH values of the equilibrium solutions were in the range of 6.2-6.5. Experimental data of the Mg adsorption onto hydroxyapatite were interpreted on the basis of a Langmuir-type model for binary systems assuming competition of Mg2+ and Ca2+ for the same adsorption sites on the crystal surfaces of the apatites.(ABSTRACT TRUNCATED AT 250 WORDS)


An active neutral metalloproteinase bound to the insoluble collagen in the mineralized phase matrix of adult rat calvaria. Two kinds of proteinases were found in the mineralized phase matrix of 24-week-old rat calvaria by means of enzymography using gelatin as a substrate. One proteinase was a neutral thiol 58kD proteinase as shown in a previous paper [2]. The other was a neutral metalloproteinase that had a molecular mass of 56kD and was detected only when calcium (Ca) ions were added to the incubation buffer. It is believed that the 56kD proteinase is bound to the insoluble collagen of the bone matrix, as it is solubilized by 4 M guanidine HC1 solution from the insoluble collagen fraction, when prepared by removing extractable proteins of the mineralized phase matrix. The insoluble collagen fraction could also be solubilized and prepared as gelatin by heating at 65 degrees C for 5 minutes. The gelatin was then incubated at 37 degrees C without further treatment and became degraded without an activation of 4-aminophenylmercuric acetate (APMA). This nonactivated degradation was enhanced by adding Ca ions. These results suggest that the 56kD metalloproteinase bound to the insoluble collagen of bone matrix is in an active form and may participate in the rapid degradation of collagen during bone resorption. As partially purified 56kD metalloproteinase degraded cartilage type proteoglycan, but not type I, IV, and V collagens, it is possibly related to the degradation of proteoglycans before it binds to collagen fibers during bone formation.


Odontogenic tumors in mice carrying albumin-myc and albumin-rats transgenes. Odontogenic tumors that produce abnormal tooth-like structures are repeatedly observed in mandibles of mice that carry both albumin-myc and albumin-ras transgenes. The earliest lesions appear among the periodontal ligament mesenchymal cells, but later lesions include an epithelial component. Subsequent tumor development recapitulates the process of normal tooth formation, which requires multiple sequential cell signals, and results in cell differentiation, matrix secretion, and mineralization. Tumor cells with epithelial morphology produce ras oncoprotein, consistent with an epithelial origin of these tumors. As albumin regulatory sequences direct oncogene expression in these mice, our findings also suggest that some of the albumin present in normal teeth may be locally produced and have a role in tooth mineral formation. The reproducibility of this phenotype makes these mice an excellent model for studies of both normal and neoplastic odontogenesis.


Ability of vertebral dimensions from a single radiograph to identify fractures. It has been proposed that vertebral dimensions be used to objectively identify vertebral fractures, permitting standardization of methodology for comparisons between studies. In this report, we evaluate the ability of various vertebral dimensions and ratios to identify "abnormal" vertebrae. As no "gold standard" exists for prevalent vertebral fractures, we examined the ability of cross-sectional dimensions (at a single point in time) to detect fractured vertebrae that had been identified from changes in dimensions compared with previous radiographs. Theoretically, a cutoff of 3 SD below the mean will rarely misclassify normal vertebrae as fractured (specificity = 99.9%). However, we found that this cutoff correctly identified only about 70% of the incident fractures. A less stringent criterion (2 SD below the mean; theoretical specificity = 97.7%) identified about 85-90% of true fractures. Dividing by stature or other vertebral heights sometimes yielded marginal improvements in the ability of the anterior or posterior height dimensions to diagnose fractures. The results suggest that the true fracture prevalence may sometimes be substantially higher than suggested by cross-sectional vertebral measurements.


Menopausal bone loss is partially regulated by dietary intake of vitamin D. Five years ago we reported results from a cross-sectional study of the effect of nutritional factors on calcium-regulating hormones and bone loss in perimenopausal women. We found an inverse correlation between serum 25-hydroxyvitamin D (25OHD) and immunoreactive parathyroid hormone (PTH), and we postulated that over time, women with lower 25OHD would lose more bone because of increased bone remodeling induced by secondary hyperparathyroidism. We have followed 38 of these women for 5 years. Twenty-two have gone through menopause and we are reporting observations on these 22 subjects. Bone mineral analysis was performed twice a year at the distal and mid-radius using single-photon absorptiometry. The slope of the bone mineral content curve was calculated by least squares. Bone loss increased within 6 months of the rise in serum follicle stimulating hormone (FSH) to greater than 40 mIU/ml. We continued to see a negative correlation between 25OHD and PTH (r = -0.450, P = 0.03). Premenopause, PTH was negatively correlated with the proximal bone mineral content (PBMC) slope (-0.604, P = 0.002). The distal bone mineral content (DBMC) 5-year slope was correlated with dietary vitamin D (r = 0.509, P = 0.02), the higher the intake, the less negative the slope. The 5-year PBMC slope was negatively correlated with serum osteocalcin (OC) levels (r = -0.382, P = 0.08). Before menopause, the change in PBMC was positively correlated with OC (r = 0.450, P = 0.03). Postmenopause, the correlation with DBMC slope was negative (r = -0.506, P = 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


Ultrasound attenuation of the calcaneus in the female population: normative data. Broadband ultrasound attenuation (BUA) on the os calcis has been proposed as a safe and reliable technique for evaluating skeleted status. The present study provides preliminary normative BUA results on 93 female subjects divided into five age groups. These data can be used as a guide for comparing the results of individual studies. The diagnosis of osteoporosis was determined from lateral calcaneus and spine radiographs. Postmenopausal osteoporotic female subjects had significantly lower BUA values than normal women (P less than 0.001). There was a significant negative linear relationship between BUA and age in the postmenopausal subjects. No correlation was found between body size parameters (weight, height, and body mass index) and ultrasound attenuation. These results indicate that BUA may be a useful technique for detection of persons at risk.


Rectal salmon calcitonin for the treatment of postmenopausal osteoporosis. In a 2-year study, we examined bone mass and calcium metabolism in 36 elderly women with moderate osteoporosis. The study period comprised 1 year of observation, during which the women received no treatment affecting calcium metabolism, and 1 year of treatment, during which all participants received daily salmon calcitonin (sCT) 100 IU rectally and calcium 500 mg. During the observational period a significant bone loss of 1.5% was seen in the forearm (P less than 0.01), whereas the spinal bone mass was virtually unchanged. After institution of treatment, the bone loss was arrested in the forearm and a significant increase of about 2% was seen in the spine (P less than 0.01). The net effect of treatment revealed a positive outcome in both bone compartments (1.9% and 2.9%, P less than 0.05-0.01). Correspondingly, the parameters of bone turnover (serum alkaline phosphatase, plasma bone Gla protein, and fasting urinary hydroxy-proline/creatinine) did not change during the observational period, but significantly declined, 10-30%, during sCT treatment (P less than 0.01-0.001). Tolerance was generally good, although in one woman, anoscopy revealed irritative changes in the rectal mucosa. We conclude that, given rectally, sCT is well absorbed and well tolerated and that it has a beneficial effect on calcium metabolism in moderately osteoporotic women.


Bone tissue response to four-month antiorthostatic bedrest: a bone histomorphometric study. A histomorphometric analysis were made on iliac crest biopsies from eight healthy male volunteers submitted to a 4-month antiorthostatic bedrest. Bone mass and bone cell parameters, reflecting resorption and formation activities, were measured before and after the bedrest period. Trabecular bone volume and mean cortical thickness were not modified despite a decreased number of trabeculae and nonsignificant increase of the trabecular thickness; total and active resorption surfaces and the number of osteoclast per mm2 of trabecular surfaces do not vary significantly. Osteoid thickness does not vary but we found a reduced osteoid surface and a nonsignificant decreased osteoid volume. Our results suggest that bone architecture may be more affected by the reduction of mechanical forces than the bone mass. These modifications were supposed to be the result of an accelerated bone turnover in the early stage of immobilization. In this study, we failed to find disuse osteoporosis; however, we must point out that the new organization of the trabeculae could affect the bone mechanical properties.


Hemopoietic functions of marrow-derived osteogenic cells. Osteoblasts, members of the marrow stromal cellular network, may play an active role in the hemopoietic microenvironment as well as in bone remodeling. In this study, we examined the extent to which marrow-derived osteogenic cells (MBA-15) possess various stromal functions. This marrow stromal-derived cell line was shown by us to exhibit osteoblastic characteristics in culture and to form bone in vivo. These cells are shown here to constitutively produce and secrete cytokines identified as M-CSF, GM-CSF, and IL-6. MBA-15 cells modulate growth of normal and malignant myeloid and lymphoid cells as well as leukemia cell lines in vitro. Cell-cell interactions were studied in co-cultures with adherent MBA-15 cells and the target hemopoietic cells. Growth inhibition effects, observed under various experimental conditions, can be attributed to the presence of different soluble and membrane-bound inhibitory activities produced by MBA-15 cells. Thus, MBA-15 cells spontaneously produce both stimulators and inhibitors that can affect myeloid and lymphoid cell growth. Marrow osteogenic cells may therefore participate in the stromal regulation of hemopoiesis.


The localization and characterization of proteinases for the initial cleavage of porcine amelogenin. In the outermost layer of porcine-developing enamel adjacent to the ameloblasts in the secretory stage, the activities of two proteinases having molecular masses of 76 and 78kDa were detected by enzymography using gelatin as a substrate. On the other hand, high activities of known 30 and 34kDa proteinases were localized in the inner layer of the enamel. The 76kDa proteinase cleaved the carboxyl-terminal peptide of porcine 25kDa amelogenin to convert it to 20kDa amelogenin. The 78kDa proteinase also acted on the 25kDa amelogenin similarly, but its activity was weak. The results indicate that the 25kDa amelogenin synthesized and secreted by ameloblasts is converted to 20kDa amelogenin by the action of proteinase localized in the outermost layer of the secretory enamel, and then further degraded by the proteinases in the inner layer of the enamel associated with the increase of mineralization.


The anabolic effect of estrogen on endosteal bone formation in the mouse is attenuated by ovariohysterectomy: a role for the uterus in the skeletal response to estrogen? In the mouse, the anabolic effect of estrogen on the uterus and its stimulatory effect on endosteal bone formation are well documented. When these observations are coupled with the recent description of uterine-derived bone cell mitogens, it raises the possibility that uterine hypertrophy in response to estrogen might lead to the production and release of factors that participate in the skeleton's anabolic response to estrogen. To determine if the stimulatory effects of estrogen on endosteal bone formation and uterine tissue in the mouse are related, we have studied this specific skeletal response to ovariectomy (OVX) and ovariohysterectomy (OHTX), and to two levels of 17 beta-estradiol (17 beta-E2). To assess treatment effects, 48 Swiss-webster mice were assigned to six groups: OHTX/oil vehicle, OVX/oil vehicle, OHTX/150 micrograms 17 beta-E2, OHTX/300 micrograms 17 beta-E2, OVX/150 micrograms 17 beta-E2, and OVX/300 micrograms 17 beta-E2. Animals were treated once per week with vehicle or the respective 17 beta-E2 dose. To quantitate bone formation, fluorochrome labels were administered at the beginning and end of the experimental period. At the conclusion of the 5-week study, tibiae were processed undecalcified for embedding in methyl methacrylate plastic. Cross-sectional areal properties and bone formation rates were quantitated from 30 microns mid-diaphyseal sections using a Bioquant Bone Morphometry system. Compared with the vehicle-treated OVX and OHTX mice, 150 micrograms of 17 beta-E2 administered once per week significantly increased cortical bone areas (P less than 0.05) but cortical bone widths and the ratio of cortical bone area to total bone area was increased only in estrogen-treated OVX mice (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)


Experimental changes in mineral content of juvenile mouse femora. Juvenile laboratory mice were exposed to hypergravity (8 g), burrowing exercise, swimming to exhaustion, an anabolic steroid, and swimming and an anabolic steroid for 30 days to determine the variability of skeletal mineralization during growth. Changes in mineralization were correlated with changes in bending strength. Experimental mouse femora were loaded to failure in a cantilever beam configuration to determine bending strength, and ashed to determine total mineral content. Between experimental groups, mineral content ranged from 66.0 to 71.2% with the greatest change from the control being a 4.7% decrease in mineralization in the male swimming exercise group (P less than 0.001). Within two age-matched experiments, the first showed that the group with the greatest decrease in mineralization also had the greatest reduction in bending strength (P less than 0.001). The second age-matched experiment showed that the group with the greatest reduction in mineralization had bending strength greatly reduced (P less than 0.001). However, in this experiment, the weakest femora were in the anabolic steroid group that did not have the mineral content reduced. We conclude that (1) mineralization of juvenile mouse femora is extremely variable given varied conditions of exercise or loading; (2) mineralization of normal bone is decreased more often and to a greater extent than increased from normal exercise controls; (3) the decrease in mineralization seen here can decrease bending strength; and (4) the decrease in mineralization seen was not caused by a decrease in a mechanical loading but was probably due to a corticosterone-mediated psychological stress response.(ABSTRACT TRUNCATED AT 250 WORDS)


Sex differences in the acquisition of total bone mineral mass peak assessed through dual-energy X-ray absorptiometry. Dual energy X-ray absorptiometry evaluation of total body bone mineral content (TBBM), total bone mineral density (TBMD), and regional bone mineral content (BMC) (head, trunk, arms, and legs) was carried out in order to assess sex differences of bone in 120 women and 121 men aged 15-29 years. Subjects from both sexes were divided into 5-year groups (15 through 19, 20 through 24, and 25 through 29 years old, respectively). Significantly higher values for TBBM, TBMD, and regional BMC were observed in males compared with females in the 20 to 24 and 25 to 29-year-old groups (P less than 0.001), but not in the group aged 15-19. After adjusting TBBM for lean body mass (LBM), we observed significantly lower values of TBBM/LBM in the males compared with females in all the age groups. A positive and significant correlation was observed between TBBM and age in the males of all the groups (r = 0.624, P less than 0.001), but not in the females. These data suggest that total bone mass peak acquisition takes place earlier in women than in men, leading to more reduced bone mass value, which in turn may be an osteoporosis predisposing factor.


Reproducibility of lateral spine scans using dual energy X-ray absorptiometry. Reproducibility of lateral spine dual energy X-ray absorptiometry (LAT DEXA) scans using a Lunar DPX-L scanner was assessed in a cadaveric phantom and in patients. One hundred phantom measurements over 7 months demonstrated a longitudinal stability of 1.7% (coefficient of variation, CV). Additional scans were performed with the phantom rotated by up to 20 degrees in each of the three orthogonal planes to assess the effects of variable patient positioning. Horizontal and vertical rotation of the spine had little effect on the estimated bone mineral density (BMD), however, axial rotation of greater than 8 degrees led to errors in the BMD measurement. One hundred consecutive patients had two lateral scans performed within 1 month. BMD (range 0.10-1.6 g/cm2) was determined for each scan by one operator. Significant overlap from ribs and pelvis was often seen with L2 and L4 vertebrae but one vertebra (L3) could be measured in every case. Intraoperator and interoperator variability was assessed by three experienced operators, each analyzing 10 patients' scans on five separate occasions, and was found to be less than 1.1% for a single vertebra. BMD estimation of vertebral bodies and midslices by lateral DEXA scans (CV% of 3.8% and 4.6%) have a 95% confidence interval of 0.074 g/cm2 and 0.096 g/cm2, respectively for two vertebrae. This variability is due mainly to axial rotation, with operator variability, horizontal rotation, and vertical rotation having little effect on BMD estimation.