Synapses on motoneuron dendrites in the brachial section of the frog spinal cord: a computer-aided electron microscopic study of cobalt-filled cells

Summary Cobalt-labelled motoneuron dendrites of the frog spinal cord at the level of the second spinal nerve were photographed in the electron microscope from long series of ultrathin sections. Three-dimensional computer reconstructions of 120 dendrite segments were analysed. The samples were taken from two locations: proximal to cell body and distal, as defined in a transverse plane of the spinal cord. The dendrites showed highly irregular outlines with many 1–2 m-long thorns (on average 8.5 thorns per 100 m² of dendritic area). Taken together, the reconstructed dendrite segments from the proximal sites had a total length of about 250 m; those from the distal locations, 180 m. On all segments together there were 699 synapses. Nine percent of the synapses were on thorns, and many more close to their base on the dendritic shaft. The synapses were classified in four groups. One third of the synapses were asymmetric with spherical vesicles; one half were symmetric with spherical vesicles; and one tenth were symmetric with flattened vesicles. A fourth, small class of asymmetric synapses had dense-core vesicles. The area of the active zones was large for the asymmetric synapses (median value 0.20 m²), and small for the symmetric ones (median value 0.10 m²), and the difference was significant. On average, the areas of the active zones of the synapses on thin dendrites were larger than those of synapses on large calibre dendrites. About every 4 m² of dendritic area received one contact. There was a significant difference between the areas of the active zones of the synapses at the two locations. Moreover, the number per unit dendritic length was correlated with dendrite calibre. On average, the active zones covered more than 4% of the dendritic area; this value for thin dendrites was about twice as large as that of large calibre dendrites. We suggest that the larger active zones and the larger synaptic coverage of the thin dendrites compensate for the longer electrotonic distance of these synapses from the soma.     

Cytodifferentiation and synaptogenesis in the neostriatum of fetal and neonatal rhesus monkeys

Summary Cytodifferentiation and synaptogenesis in the neostriatum (caudate nucleus and putamen) were analyzed by the Golgi impregnation method and electron microscopy in 14 fetuses and 8 postnatal rhesus monkeys. During the second fetal month the neostriatum consists primarily of simple, mostly bipolar, immature cells and a small number of undefined profiles ending with growth cones. The first morphologically defined synapses appear in the putamen at embryonic day 60 (E60) and in the head of the caudate nucleus at E65. Synaptic density in both structures is less than one per 1000/m² of neuropil at this stage; synapses are characterized by asymmetric junctions between axonal profiles and immature dendritic shafts, accumulation of an intermembrane web and aggregation of round clear vesicles in presynaptic profiles. During the third fetal month neuronal cell bodies and glial cells enlarge, and axonal and dendritic processes in Golgi preparations become more complex. Although the basic morphology of synapses remains unchanged, their density increases to 9/1000 m² in the putamen and 3.7/1000 m² in the caudate. During the fourth fetal month the four principal cell classes of the neostriatum emerge. Spines on the shafts of dendrites are followed closely by the appearance of axospinous synapses. Synaptic density in the putamen is still significantly higher (10.1/1000 m²) than in the caudate (5.4/1000 m²), but by the end of the fifth fetal month (E150) it is the same (80/1000 m²) in both structures. A dramatic increase in synaptic density to 125/1000 m² occurs before term (E165) with the emergence of the first asymmetric synapses as well is symmetric synapses with flat or pleomorphic vesicles that terminate predominately on dendritic shafts. Synaptic density continues to increase after birth, reaching a plateau of approximately 190/1000 m² at the end of the first postnatal month. Throughout postnatal development the proportions of symmetric and asymmetric synapses on the smooth dendritic shafts undergo systematic fluctuations which may reflect the ingrowth of various afferents as well as local cytological differentiation including the formation of cellular compartments.     

Mechanism of modulation of single sodium channels from skeletal muscle by the β 1-subunit from rat brain

We studied the molecular mechanism of the rat skeletal muscle -subunit (_I) gating kinetics modulation by the brain ₁-subunit by heterologous expression of single sodium channels from _I and ₁ in Xenopus laevis oocytes. Coexpression of ₁ reduced mean open time at –10 mV to 21% when compared to channels expressed by _I alone. Channels formed by _I exerted multiple openings per depolarization, which occurred in bursts, in contrast to the channels formed by the _I/₁ complex that opened in average only once per depolarizing voltage pulse. Macroscopic current decay (mcd), as evidenced by reconstructed open probability vs. time , was greatly accelerated by ₁, closely resembling mcd of sodium currents from native skeletal muscle. Generally was larger for channels expressed from the pure _I subunit.From our single channel data we conclude that ₁ accelerates the inactivation process of the sodium channel complex.     

Modulation of lymphocyte activating factor activity (interleukin 1 like activity) and acute phase proteins in pertussis-induced air pouch inflammation by muramyl dipeptide

We have studied the effect of the macrophage activator, muramyl dipeptide (MDP) on immune inflammation induced in the rat six day subcutaneous air pouch. Treated animals received either 100 g or 200 g MDP at the time of challenge and twenty four hours before exudate harvest. Using the thymocyte co-mitogenic assay for lymphocyte activating factor (LAF), 100 g MDP enhanced LAFactivity whereas 200 g caused inhibition. Increased dilution of 200 g exudate in this assay removed this inhibition. Similarly, at the lower dose, MDP caused enhanced production of the acute phase protein alpha 1 glycoprotein, whereas the higher dose had no effect. The present study suggests that macrophage activity can be manipulatedin vivo to produce LAF and naturally occurring inhibitors of LAF. These studies indicate that the stimulation of LAF inhibitors by MDP may be a potential theerapeutic action.     

Enhanced stability of YEp plasmids in lager brewing yeasts is related to lager brewing yeast 2-μm DNA

Summary YEp plasmid stability in the presence of either Saccharomyces cerevisiae laboratory strain 2-m DNA, or lager brewing yeast 2-m DNA in the same genetic background, was compared under non-selective culture conditions. It was found that YEp plasmids were more stably maintained in the presence of lager 2-m DNA under these conditions. By construction of laboratory-lager 2-m DNA hybrid plasmids, an 867 bp StuI fragment of lager 2-m DNA was shown to be responsible for the enhanced stability of the YEp plasmid. Nucleotide substitutions at two sites were found by sequencing this region. It was also confirmed that increasing cell ploidy enhanced YEp stability under non-selective conditions.     

Dye coupling between dorsal raphe neurones

Neurones in the dorsal raphe nucleus (DRN) were impaled and filled with biocytin in coronal slices of midbrain taken from young adult rats. The electrophysiological properties and gross morphology of the cells were similar to those reported previously for serotonergic neurones in the DRN. Of 27 cases in which filled neurones were recovered in histological material, almost half (48%) showed labelling of two or three cells, although only one cell had been recorded from. Coupled cells were identified as close or distantly coupled, depending on the distance from the soma of the presumed impaled cell (23.5±15 m, n=7 and 150±26.5 m, n=10 respectively). Whereas close-coupled cells may have been artefactually coupled by the penetrating electrode, coupling between distant cells is most likely to be a result of transfer of biocytin through gap junctions. Camera lucida reconstructions of pairs of labelled cells revealed extensive overlap of dendritic fields and numerous crossings between dendrites. When examined at high magnification under a light microscope, many of the crossing dendrites were found to travel in different focal planes. Nevertheless, for each pair of cells, at least one point of close apposition was observed between dendrites or between the axon and a dendrite of the presumed impaled and coupled cell. The incidence of dye coupling between neurones in the DRN may reflect a relatively high level of electrotonic coupling between the neurones. This form of coupling may be important in determining the synchronous nature of firing of neurones in the DRN.     

Light microscopic and three-dimensional morphology of the human muscular sarcocyst

Established criteria for morphological typing of sarcocysts was applied to a large series of cases of human skeletal muscle sarcocystosis in Malaysia to determine the type of sarcocyst present. We also wanted to test the general usefulness of this classification and to determine if there are any new cyst types. Three-dimensional (3-D) reconstruction was done to see if the sarcocyst has a distinct 3-D morphology. A total of 66 sarcocysts from 21 cases of human muscle sarcocystosis obtained from a previous prevalence study were examined. Tissue sections (5 m thick) were stained with haematoxylin and eosin and studied under the light microscope. For 3-D reconstruction, an, image analyser was used to align and reconstruct the sarcocyst after microscopic images had been captured with a charge-coupled device (CCD) camera. All the cysts best fit into the type 4 category. This classification is generally useful, although cyst wall characteristics and zoite size appear to be the most reliable criteria for classification. The cyst width averaged 77 m (range, 30–137.5 m). Cyst walls were smooth, had no cytophaneres and were less than 1 m thick. No secondary cyst wall or surrounding inflammation was evident. Numerous cyst merozoites with diameters averaging 1 m filled the cyst lumen. Although septa were not apparent, in many cysts, zoites were arranged in a unique, curvilinear fashion that suggested their presence. 3-D reconstruction showed the sarcocyst to be a long, tortuous cylinder with no branching or other distinguishing feature.     

A current source density analysis of field potentials evoked in slices of visual cortex

Summary The method of one-dimensional current source density (CSD) analysis was applied to field potentials recorded from 350 m thick slices of the primary visual cortex of rats and cats. Field potentials were elicited by stimulation of the white matter and recorded along trajectories perpendicular to the cortical layers at spatial intervals of 25 to 50 m. The resulting CSD distributions resembled closely those recorded from the cat visual cortex in vivo. The responses with the shortest latency were distinct sinks in layers IV and VI probably reflecting monosynaptic EPSP's from specific thalamic afferents. From layer IV activity was relayed along three major routes: 1. to the supragranular layers via strong local connections to layer III and from there via both short and long range connections to layer II, 2. to targets within layer IV, and 3. to layer V. The source distributions suggest that the projections to layers III and II terminate on the proximal and distal segments, respectively, of apical dendrites of layer III pyramidal cells while the projection to layer V contacts the apical dendrites of layer VI pyramidal cells. These results indicate that all the excitatory pathways that are detectable with the CSD technique in the in vivo preparation remain intact in 350 m thick cortical slices. However, in the slice paired pulse stimulation did not lead to a depression of the response to the second stimulus while this is the case in vivo. This might be due to reduced inhibition in the slice which has been reported by several authors.     

Anatomy of soleus α-motoneurone dendrites in normal cats and in cats subjected to chronic postnatal tenotomy or overload of the soleus muscle

Summary The anatomy of intracellularly HRP-labeled soleus -motoneurone dendrites was studied both in normal adult cats (normal soleus, NS) and in adult cats which at a postnatal age of 5–7 days had been subjected to chronic tenotomy of either the soleus muscle (tenotomized soleus, TS), or all the soleus synergists contributing to the achilles tendon (overloaded soleus, OS). A set of structural rules seemed to govern the architecture of normal soleus -motoneurone dendrites. Thus, the dendrites branched dichotomously and the number of daughter branches originating from a preterminal branch was proportional to the diameter of that parent branch. Branch diameter decreased across branching points according to the 3/2 power rule of Rall (1959). Branching occurred down to a preterminal branch diameter of about 0.8 m. Through all branch orders there existed a quite precise relation between the diameter of a preterminal branch and the membrane area of its distal dendritic arborization. The average dendritic path distance from soma to termination was not closely related to the diameter of the stem dendrite, since thick stem dendrites rather generated more profusely branched arborizations than thin stem dendrites. As a corollary of these characteristics close relations existed between the dendritic stem diameter on one hand, and the total number of branches, combined dendritic length, total dendritic membrane area and total volume, on the other. In the OS material, the dendrites were not different from those of normal soleus motoneurone dendrites. In the TS material, the dendrites were less branched and had greater dendritic path lengths, although the relations between various size-parameters within the dendrites were not significantly altered compared with normal dendrites. It was concluded that the change in branching pattern was due to a net elimination of dendritic branches following the muscle tenotomy.     

Ultrastructure of a large ventro-lateral dendritic bundle in the rat ventral horn

Summary There is an extensive bundle of dendrites with a rostro-caudal axis in the ventro-lateral lamina of the sixth lumbar segment in each side of the rat spinal cord. Such a bundle has a diameter of about 250 m and contains over 1400 parallel dendrites., each with a diameter of less than 8 m, interspersed between neuronal somata. The volume fraction of dendrites in the bundle neuropil is about 55%, the remainder being equally distributed between astrocytes, synaptic boutons, and axons, most of which are unmyelinated. An analysis of the median percentage covering of dendrites by contiguous elements of the neuropil reveals that as the dendrite diameter decreases from 4 to 0.2 m (mean=2 m), astrocytes increase from 43 to 75%, axons decrease from 21% to zero, boutons decrease from 28% to zero, and dendrites decrease from 10% to zero. There is a mean of 18 synaptic boutons per 100 n² of the overall dendritic surface, but larger boutons tend to be more frequent on larger dendritic profiles. Apposed dendrites and their somata may have either puncta adhaerentia or confronting subsurface cisternae. Synaptic types in the rat are similar to those reported for the cat. The morphological findings are discussed with respect to previously proposed interaction between neural elements.     

Guanosine metabolism in adult rat cardiac myocytes: ribose-enhanced GTP synthesis from extracellular guanosine

The metabolic fate of transported guanosine was examined in adult rat cardiac myocytes. Freshly isolated cells were incubated with 10 M or 100 M [³H]guanosine and the nucleotide products extracted and examined for radiolabel distribution. The data presented show significant incorporation of guanosine into the 5-nucleotide pool, and a marked stimulation of that incorporation by ribose. An average of 233 pmol/mg cell protein extracellular guanosine was incorporated into the cellular 5-nucleotides over 90 min at both 10 M and 100 M external nucleoside. This appeared primarily as GTP (approx. 204 pmol/mg cell protein in 90 min). Only guanine nucleotides contained radiolabel; adenine nucleotides and IMP remained unlabelled even after 90 min incubation of the cells with [³H]guanosine. Addition of 5 mM ribose to the medium stimulated guanosine incorporation into 5-nucleotides 1.6-fold (380 pmol/mg protein vs 234 pmol/mg over 90 min at 10 M guanosine), but did not enhance the amount of guanosine transported into the cells. Intracellular guanosine concentrations exceeded those of the incubation medium at both external guanosine concentrations studied. More [³H]guanosine was salvaged at 100 M than at 10 M external guanosine (562 vs 380 pmol/mg protein in 90 min), but only if ribose was present in the medium. We conclude from these studies that guanosine is salvaged by heart muscle, and that at high guanosine levels the rate of guanosine salvage appears dependent on the availability of phosphoribosylpyrophosphate within the cells. At lower guanosine levels in the presence of ribose, cell guanine concentrations limit the rate of guanosine incorporation into 5-nucleotides.     

Introduction of nonselectible 2μ plasmid into [cir°] cells of the yeast S. cerevisiae by DNA transformation and in vivo site-specific resolution

Summary The 2 DNA plasmid of the yeast Saccharomyces cerevisiae does not confer any known selectable phenotype to the host cell carrying it. Selection of cells transformed with purified 2 DNA therefore cannot be achieved, and the intracellular presence of 2 can only be assessed by molecular analysis of the DNA complement. In addition, 2 alone does not replicate in bacterial hosts, thus rendering its amplification by conventional methods impossible. We have isolated a shuttle plasmid, pBH-2L, generated by in vivo sites-pecific recombination between the endogenous 2 DNA plasmid and pRL, a pBR322 derivative containing the yeast LEU2 gene and one 2 repeat sequence associated with the origin of replication. This new shuttle plasmid has the property, when transformed into yeast, of undergoing site-specific recombinational resolution between its two direct repeat sequences. This releases 2 plasmid and pRL as individual molecules. The latter can undergo progressive mitotic loss during growth in nonselective medium, ultimately leaving leucine auxotrophic transformants that contain only 2 DNA plasmid. This system can be utilized to introduce 2 DNA alone into cells lacking it, thereby providing a novel means to study the biology and the molecular genetics of the plasmid and its potential practical applications as a vector.     

Light microscopical study of dendrites and perikarya of interneurones mediating la reciprocal inhibition of cat lumbar alpha-motoneurones

Summary Interneurones which mediate disynaptic inhibition from la muscle spindle afferents of the quadriceps nerve to lumbar alpha-motoneurones were stained with intracellular injection of horseradish peroxidase. Seven best stained and most satisfactorily preserved cells were selected for analysis, and the light microscopic morphology of their cell bodies and dendrites were quantitatively investigated in parasagittal sections. The perikarya were located dorsal or dorso-medial to the motoneurones; they had mean diameters of 51 × 27 m and a mean volume of 35820 m³. The cells had 3 to 7 dendrites, which were arranged asymmetrically around the parent somata. The dendrites extended mainly in the dorso-ventral direction, in which the mean tip to tip distance for each cell was 1742 m. The dendrites had few spines and they branched almost only in bifurcations. On the average, each process divided 3.5 times and in each cell they gave rise to 14.9 branching points as well as a total combined length of more than 7000 m. Primary dendrites had a mean length of 193 m which was generally shorter than the lengths of the branches of higher order. A more detailed analysis of two cells revealed the mean width of primary dendrites to be 5.6 m while that of the 5th order processes was 1.5 m. The mean tapering of individual dendritic branches per unit length was 17%, being somewhat more pronounced for the distally located segments, while at branching points the sum of daughter processes approximately equalled the diameter of the parent process. The surface area and volume of the dendrites constituted 90% and 83% of the total surface area and 46% and 37% of the total volume of the two cells, respectively, excluding the axons. The Ia interneurones differed considerably among themselves with respect to the quantitively investigated parameters. They resembled the inhibitory Renshaw cells of the cat with regard to the number of dendrites, the poverty of spines, and the relationships between cell body diameter and width of primary dendrites.     

Effect of auranofin,a new antiarthritic agent,on immune complex-induced release of lysosomal enzymes from human leukocytes

Auranofin, an oral chrysotherapeutic agent effective in the treatment of rheumatoid arthritis (RA), was found to be a potent, noncytotoxic inhibitor of IgG-RF immune complex-induced lysosomal enzyme release (LER) from human leukocytes. At a concentration of 1 g Au/ml (5M), auranofin produced a marked reduction in-glucuronidase (100%), acid phosphatase (88%), and lysozyme (72%) release. In contrast, gold sodium thiosulfate (GST, an injectable gold compound) had no inhibitory activity on LER at equivalent gold concentrations (i.e., 1g Au/ml) and only modest activity (< 36% inhibition) at concentrations as high as 40g Au/ml. The 50% inhibitory dose (ID₅₀) of auranofin on LER was calculated to be 3–4M (0.6–0.8g Au/ml). Blood gold levels in auranofin-treated RA patients were found to be within the range required for in vitro inhibition of LER, and correlated with decreases in IgG, RF titers, and IgG-RF immune-complex formation in vitro. These results suggest that the therapeutic action of auranofin may be caused, at least in part, by inhibition of LER and/or decreases in immune-complex formation.SK&F D-39162 (2,3,4,6-tetra-O-acetyl-1-thio--D-glucopyranosato-S) (triethylphosphine) Gold.     

Präzisierung der Reizwirkung mittelfrequenter Wechselströme

Zusammenfassung Bei der Mittelfrequenz-Impuls-Reizung ist streng darauf zu achten, daß keine polaritären Reizkomponenten auftreten. Die diesbezügliche Kontrolle wird am besten mit Hilfe des Konvertibilitätstestes vorgenommen, d. h., es darf beim Vertauschen der Zuführungen zu den Reizelektroden weder die Reizschwelle bzw. die Größe des kollektiven Reizerfolges noch dessen Latenzzeit eine signifikante Änderung erfahren. Auf diese Weise wird die Phasenunabhängigkeit des echten Mittelfrequenz-Reizeffektes nachgewiesen.Diesen Anforderungen entsprechen Mittelfrequenz-Impulse, deren Trägerfrequenz über einige wenige Perioden sich aufschaukelt und ebenso wieder abklingt. Demgegenüber sind Mittelfrequenz-Stromstöße mit phasenstarrem Einsatz und Ende nicht unbedingt frei von polaritären Ein- bzw. Ausschalt-effekten, indem sowohl die erste als auch die letzte Trägerperiode einen polaritären Wechselimpuls-Reizeffekt ergeben kann, je nach Phasenlage bezogen auf die wirksame Reizelektrode und Art der Ansprechbarkeit des Reizobjektes (Nerv) auf entsprechend kurze gleitspiegelsymmetrische Wechselimpulse. Für eine echte Mittelfrequenz-Stromstoß-Reizung ist demnach ebenfalls ein Aufschaukeln und Abklingen der Trägerfrequenz über einige wenige Perioden erforderlich.Es besteht ein prinzipieller Unterschied zwischen der echten Mittelfrequenz-Reizung, die phasen -bzw. periodenunabhängig ist und schon früher als apolaritär bezeichnet wurde, und der konventionellen polaritären Reizung, die als polaritäre Komplikation der Mittelfrequenz-Reizung auftreten kann.Diese Präzisierung der Reizwirkung mittelfrequenter Wechselströme wurde angeregt durch zwei im Text erwähnte Publikationen, in denen in keineswegs überzeugender Weise versucht wird, die Mittelfrequenz-Reizung letzten Endes auf das polare Gesetz der Erregung zurückzuführen.

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Summary The particular excitatory action exerted by middle-frequency alternating current can only be revealed if care is taken to eliminate the occurrence of so-called polarity effects. Such effects are produced by the short alternating impulses represented by the first and the last period of a middle-frequency current pulse and are based on the polar law of excitation.In order to prevent such polarity intrusions, it is necessary to increase and decrease the amplitude of the middle-frequency current pulses over a few carrierperiods, or, to use amplitude-modulated middle-frequency impulses of variable shape and duration of envelope.A true middle-frequency excitatory effect is easily demonstrated by resorting to the convertibility test. It will then become evident that stimulation threshold, magnitude as well as latency of response do not change during reversal of the stimulating poles. This means, that no significant phase change of the response with regard to the carrier-frequency occurs when the leads to the stimulating electrodes are commuted, and that, as a result, true middle-frequency effects do not depend upon one particular catelectrotonic variation among the carrier-periods of a middle-frequency current pulse.It can thus be concluded that a fundamental difference exists between true middle-frequency stimulation, which is based on a non-polarity or apolarity principle, and the conventional stimulation of the polar or polarity type.This paper has been written in the hope of dispelling some errors of interpretation (discussed in the text) tending to ascribe the excitatory effects of middle-frequency impulse stimulation to the classical polar law of excitation.

     

Activity of trimethoprim/ sulfamethoxazole plus polymyxin B against multiresistantStenotrophomonas maltophilia

The inhibitory activity of eight antibiotics and the inhibitory and bactericidal activities of combinations of trimethoprim/sulfamethoxazole (TMP/SMX) plus three fixed concentrations of polymyxin B (0.01 g/ml, 0.1 g/ml and 0.5 g/ml) against 30 multiresistant strains ofStenotrophomonas maltophilia were tested. Polymyxin B at 0.01 g/ml modified the inhibitory activity of TMP/SMX against only 40% of strains. At 0.1 g/ml and 0.5 g/ml, polymyxin B enhanced the inhibitory activity of TMP/SMX activity against all strains. Polymyxin B enhanced the bactericidal activity of TMP/SMX only at concentrations near the minimum inhibitory concentration of polymyxin B alone.     

Synaptic extensions from the mossy fibers of the fascia dentata

Summary In the hilar region of the rat hippocampus, filamentous extensions have been observed to originate from the en passant synaptic expansions on the so-called mossy fibers, which are the axons of the dentate granule cells. These extensions range in length from about 1 m to 30 m, are often branched, and appear to contact the processes of various cell types in the hilar region. In 28-day-old rats, there are between 4 and 9 such extensions from most mossy fiber expansions, and the total length of the extensions from any one expansion is on the order of 75 m. Analysis of serial electron micrographs through normal and Golgi-impregnated mossy fibers has confirmed that these extensions are, indeed, presynaptic processes. Each contains one or more vesicle-rich foci along its length, and is associated with asymmetric membrane specializations. At these sites, the extensions are in synaptic contact with dendrites and dendritic spines of, as yet, unknown origin. A quantitative analysis of these extensions in Golgi material from rats at different ages indicates that they reach their greatest length around 14 days and then decline to adult values by 28 days.This work was supported in part by grant NS-10943 and was carried out while the author was in receipt of a U.S. Public Health Service post-doctoral fellowship (F32 NSO 5765-02)     

Differential regulation of human eosinophil,macrophage, and neutrophil functions by the allergic mediator release inhibitor CI-959

The cell activation inhibitor CI-959 (5-methoxy-3-(1-methyl-ethoxy)-N-1H-tetrazol-5-ylbenzo[b]thiophene-2-carboxamide, monosodium salt) was evaluated for its effect on the activation of human eosinophils, macrophages, and neutrophils by the phagocytic stimulus serum-opsonized zymosan (SOZ). CI-959 inhibited the respiratory burst of eosinophils and neutrophils, measured as the generation of superoxide anion, with IC₅₀s of 9.6 and 14.5 M, respectively. In contrast, 100 M CI-959 inhibited superoxide anion generation by human macrophages by only 22.7%. The compound exhibited a different inhibition profile for lysosomal enzyme release from these cells. At 100 M, CI-959 inhibited the release of eosinophil peroxidase and macrophageN-acetyl--d-glucosaminidase by only 19.5 and 25.6%, respectively. In contrast, CI-959 inhibited the release of the neutrophil primary granule enzyme myeloperoxidase with an IC₅₀ of 7.5 M, while inhibiting release of lysozyme from secondary granules by only 11.4% at 100 M. These results demonstrate that oxygen radical generation and lysosomal enzyme release by human leukocyte populations are differentially regulated by CI-959.     

Susceptibility to beta-lactam antibiotics in septicemia isolates from twenty-nine European laboratories

In 1984 the European Study Group on Antibiotic Resistance (ESGAR) consecutively collected gram-negative bacilli and staphylococci blood isolates and performed susceptibility testing with 11 antibiotics using the microdilution method. In all 2,578 isolates were collected: 68% gram-negative bacilli and 32% staphylococci. The MICs of ampicillin and cefazoline for the susceptible gram-negative bacilli were 1–8g/ml; of piperacillin0.5–4; of Sch 34343, cefotaxime, moxalactam, ceftazidime and aztreonam0.5–2g/ml; of cefoxitin, cefuroxime and cefamandole0.5–8g/ml. For susceptible staphylococci the MICs of cefazoline and cefuroxime were0.5–1g/ml, and of cefoxitin, moxalactam, ceftazidime and cefotaxime,0.5–32 g/ml. The resistance levels varied between laboratories and countries, being lower in Northern Europe. In clinical protocols on patients with gram-negative septicemia from whom cefazoline-resistant strains were isolated, cefotaxime was the beta-lactam most commonly used (12%). In protocols on patients with staphylococcal septicemia from whom gentamicin-resistant or cefazoline-resistant strains were isolated, the most commonly used beta-lactam was cloxacillin (6%).     

Scanning electron microscopy of merogonous stages ofEimeria falciformis var.pragensis inMus musculus

Asexual stages ofEimeria falciformis var.pragensis in Swiss-Webster mice were studied by scanning electron microscopy. Sporozoites were present in the cecum and colon 2 h post-inoculation (PI) and measured 11.3×2.1 m (9–13.9×2–2.2 m). Sporozoites penetrated epithelial cells with an extended anterior end and were constricted at the site of entry. Asexual generations were found in the cecum and colon epithelial cells. In meronts found at days 3–6 PI, merozoites matured synchronously, were oriented in the same direction, and were arranged in a helical pattern. Such meronts measured 11.3×6.4 m (8–13.7×5–7.4 m) and contained 8–12 meroxoites, which measured 11.9×1.5 m (7.4–15.7×1.3–1.8 m). Meronts which were present at day 7 PI measured 9.5×8.2 m (9–10.5×7–9.5 m) and contained 20–50 small merozoites which budded asynchronously from a central residuum. At days 3–7 PI, parasitized epithelial cells had shorter and fewer or no microvilli. The lumenal plasmalemma of the host cell was often disrupted or absent in cells containing mature meronts and escaping merozoites. At day 6 PI, phagocytic cells appeared on the epithelial surface, some of which were in contact with merozoites. Small foci of exposed basal lamina were present at day 7 PI in areas where cells had sloughed from the epithelium.