Gangliosides and Glycosphingolipids of Peripheral Nervous System Myelins—a Minireview

A summary is provided of the available data on the composition of gangliosides and glycosphingolipids in the peripheral nervous system (PNS) including myelins and their antigenic properties. The composition of gangliosides and glycosphingolipids in the PNS is very different from that in the central nervous system (CNS), both quantitatively and qualitatively. One major difference is the abundance of neolacto-series gangliosides in the PNS, with the backbone structure Gal 1-4GlcNAcl-3Gal l-4Glcl-lCer. Their abundance contrasts with the abundance of ganglio-series gangliosides in the CNS. The neolacto-series gangliosides are localized mainly in the myelins of the PNS. In addition to gangliosides, other acidic and neutral glycosphingolipids in the neolacto-series are also characteristic of the myelins of the PNS. The ceramide (fatty acid and sphingosine base) compositions of gangliosides in the PNS are different from those in the CNS gangliosides, having greater percentages of long-chain fatty acids and dehydrosphingosines than found in the CNS gangliosides.     

Über das Wachstum der Pankreaszellen von Säugetieren als Monolayer Cultures

Zusammenfassung Fetale menschliche Pankreaszellen bilden sowohl spontan auf zuerst gewachsenem Zellrasen (self-aggregation) als auch in Rollröhrchen (48 rph) (rotation-mediated-aggregation) histotypische Aggregate, die A- und B-Zellen enthalten. In der Nährlösung dieser 6–17 Tage lang gewachsenen Gewebestückchen konnte, Insulin (=IRI=immunoreactive-insulin) nachgewiesen werden.

---

Monolayer cultures of pancreatic tissueII. Formation of histotypic aggregates from cell suspensions of fetal human pancreas

Summary Fetal Pancreas cells of man, whether in self-aggregation or rotation-mediated-aggregation, form spontaneously histotypical aggregates containing A- and B-cells. It is possible by means of an immunological method to determine insulin (=IRI=immuno-reactive-insulin) in the nutrient medium of these 6 to 17 days old tissue aggregates.

     

Potentiation of growth suppression and modulation of the antigenic phenotype in human melanoma cells by the combination of recombinant human fibroblast and immune interferons

Summary Administration of interferon as a single therapeutic regimen in cancer patients with various neoplasias has had only limited efficacy in ameliorating the negative clinical course of their disease. In the present study, we have evaluated the effect of recombinant human fibroblast (IFN) and immune (IFN) interferon, alone and in combination, on growth, differentiation and the expression of class I and II histocompatibility locus antigens (HLA) and melanoma-associated antigens on the human melanoma cell line H0-1. The effect of combinations of interferons on the antigenic profile of human melanoma cells displaying different organ colonization and spontaneous metastatic potential in athymic nude mice was also determined. H0-1 cells were more sensitive to the antiproliferative activity of IFN than to IFN and the combination of interferons resulted in a potentiation of growth suppression. The antiproliferative effect of both interferons was greater in later-passage than in earlier-passage H0-1 cells, possibly reflecting alterations in the evolving tumor cell population as a result of long-term in vitro propagation and/or the selective outgrowth of cells with an increased growth rate. The enhanced growth suppression observed in H0-1 cells treated with the combination of IFN plus IFN was not associated with a significant increase in the level of melanin, a marker of melanoma differentiation, above that observed with either interferon used alone. IFN and IFN differentially modulated the expression of class I and II HLA and melanoma-associated antigens in H0-1 cells and a series of melanoma cells with different organ colonization and metastatic potential, including MeWo, MeM 50-10, MeM 50-17, 3S5 and 70W. No consistent potentiation or antagonism in the expression of any specific antigen was observed in any of the melanoma cell lines exposed to the combination of interferons. The present study demonstrates that the combination of IFN plus IFN can potentiate growth suppression in H0-1 human melanoma cells and that this effect is not associated with an increase in differentiation or a potentiation in antigenic modulation. In addition, no direct correlation between the expression of any specific antigen or its modulation by IFN or IFN, alone or in combination, and organ colonization and metastatic potential in nude mice was observed in the different melanoma cell lines.     

Molecular characterization of β-thalassemia in Czechoslovakia

Summary We have identified different -thalassemia mutations in 93 members of 34 families of Czech or Slovakian descent using gene amplification, hybridization with specific ³²P-labeled oligonucleotide probes, sequencing of amplified DNA, and gene mapping. The GA mutation at IVS-I-1 was found in 18 families; other Mediterranean mutations were IVS-II-1 (GA), IVS-II-745 (CG), IVS-I-110 (GA), and codon 39 (CT); these were present in 9 additional families. The GT mutation at codon 121, known to cause Heinzbody -thalassemia, was present in 3 families, and the frameshift at codons 82/83 (-G), first described in the Azerbaijanian population, in 2 families. A newly discovered allele was a frameshift at codons 38/39 (-C). One -thalassemia allele was incompletely characterized. We observed in 2 families a TC mutation at position +96 UTR (untranslated region) relative to the termination codon; this mutation likely is a rare polymorphism, -Thalassemia was rare; only one person carried the -^3.7 heterozygosity, and one other had a yet to be identified -thalassemia-1, while seven had the ^{anti 3.7} triplication.     

Cytochemical localization of β-galactosidase in resident and inflammatory peritoneal macrophages from C57BL mice

Summary A cytochemical method for the detection of -galactosidase (-Gase) in mouse peritoneal macrophages was used to study the ultrastructural localization of this enzyme in these cells. It was found that the reaction product for -Gase was localized in the perinuclear cisternae, the endoplasmic reticulum, the Golgi complex, lysosomes, vesicles and on the cell surface of peritoneal macrophages from untreated C57BL mice. When examined by X-ray microanalysis the crystalline reaction product was found to contain bromine, an element present in the indolyl substrate which was used to identify -Gase. Injection of Proprionibacterium acnes (P. acnes) intraperitoneally or BCG intravenously caused a visible loss in -Gase from all the organelles and from the cell surface of the macrophages.Abbreviations used -Gase -galactosidase - RP reaction product - PNC perinuclear cisternae - RER rough endoplasmic reticulum     

Colony cycle of an Australian swarm-founding paper wasp,Ropalidia romandi (Hymenoptera: Vespidae)

Summary Colonies of an Australian swarm founding polistine wasp,Ropalidia romandi, on the Atherton Tableland, Queensland, manage perennial nests by practicing seasonal colony activity. In summer nests, most cells were occupied by immatures of all developmental stages. The proportion of cells that did not contain immatures increased towards winter, when most nest cells did not contain immatures and half of these winter nests had rather large amounts of nectar deposited in empty cells. While the visits to tree flowers byR. romandi were not frequent in summer, nearly 60 % of insects collected on tree flowers in winter wereR. romandi. With such seasonal switching in the modes of colony reproductive activity and of nectar foraging behavior,R. romandi responds to its physical and biotic environment. In this way,R. romandi may be able to continue colonies throughout the year, with much reduced level of colony activity in winter, in areas with seasons that are unfavorable for reproductive activity of a colony.     

Restriction fragment analysis of the secalin loci of rye

Analyses of wheat/rye addition lines by Southern blotting confirmed the presence of sequences related to theSec 1, Sec 2, andSec 3 loci on chromosomes 1R and 2R. Comparison of the 1R and 2R addition lines allowed the identification of -secalin genes atSec 1 andSec 2, respectively, while -secalin and -secalin genes atSec 1 were discriminated by comparative hybridization with three probes: -secalin, total -secalin, and 3 -secalin. The high molecular weight (HMW) secalin genes atSec 3 were identified using a homologous HMW subunit probe from wheat. Gene copy numbers were estimated as about 40–60 for -secalins, 5–10 for -secalins, and 2 for HMW secalins. Comparison of individual plants of cv. Gazelle showed a high degree of polymorphism, particularly for sequences related to -secalins and HMW secalins.     

Synaptic junction development in the spinal cord of an amphibian embryo: An electron microscope study

Summary An electron microscopical study has been made of the cervical spinal cord of Xenopus laevis embryos, from the time that the neural tube closes until the larvae were hatched and could swim. Sections of the whole cord were searched for intercellular junctions during this period. Two nonsynaptic types were found, the first were widely distributed puncta adherentia, the second were rare and similar to gap junctions. Membrane specializations with synaptic vesicles were first found when the neural folds had fused; membrane-vesicle clusters which looked like the presynaptic half of a synaptic junction were present, together with synaptic junctions lacking any postsynaptic membrane thickening or cytoplasm density. About four hours later, mature synaptic junctions with full thickening of the postsynaptic membrane, dense cytoplasm and striated or dense material in the synaptic cleft were present. Presynaptic mitochondria, dense-cored and flattened vesicles, fibre to fibre and fibre to cell body synapses were present from the first, as were synapses onto very fine dendrites which might be filopodia from dendritic growth cones. Synaptogenesis may start with the accumulation of vesicles in dense cytoplasm near a thickened cell membrane; the postsynaptic element becomes associated with this membrane-vesicle cluster and matures by increasing cleft and cytoplasmic density, and by membrane thickening.     

Gonadectomy induces laminin biosynthesis and basement membrane assembly in anterior pituitary glands of adult rats

Summary Laminin biosynthesis and basement membrane assembly in anterior pituitary glands of gonadectomized rats were studied by immuno-electron microscopy and radioimmunoassay. Three weeks after gonadectomy, rats received intravenous injections of sheep anti-laminin IgG conjugated to horseradish peroxidase, and glands were fixed and processed for microscopy 1 h later. Peroxidase reaction product uniformly labeled all perivascular and glandular epithelial basement membranes. In addition, reaction product was also found in abnormally multi-layered basement membranes seen especially beneath gonadotrophs, and unusual basement membrane-like structures projecting between gonadotrophs were also labeled. Pituitary sections from gonadectomized rats labeled with pre-embedding immunoperoxidase and post-embedding immungold techniques also localized intracellular laminin within biosynthetic organelles and light body vesicles of gonadotrophs. Neither abnormal basement membrane structures nor intracellular laminin were detected in pituitaries of nongonadectomized, control rats. Radioimmunoassays of pituitary homogenates showed nearly twice as much soluble laminin ( 15 ng/gland) in gonadectomized rats than in controls ( 8 ng/gland), which paralleled gland growth, but serum laminin concentrations did not differ ( 10 ng/ml in both groups). When anterior pituitary glands of gonadectomized rats that received injections of anti-laminin IgG-HRP were fixed 5 days after injection, lengths of unlabeled basement membrane were distributed between labeled lengths. This indicated that new basement membrane was spliced into old by a process similar to that seen in normal development. Supplementation of gonadectomized rats with testosterone, however, arrested laminin biosynthesis and basement membrane assembly and reversed glandular hypertrophy. These results indicate that, in an absence of sex hormone feedback, renewed synthesis of basement membrane components occurs in the anterior pituitary and is probably necessary to support the additional growth and differentiation of gonadotrophs and other pituitary cells.     

Cluster Maintenance in Mobile Ad-hoc Networks

The main contribution of this paper is to propose a new cluster maintenance algorithm and a companion cluster initialization algorithm based on a number of interesting and novel properties of diameter-2 graphs. The initialization algorithm naturally blends into cluster maintenance, showing the unity between these two operations. We refer to our algorithms as tree-based since they depend on a spanning tree maintained at various nodes. Unlike the vast majority of published clustering algorithms, our algorithms are cluster-centric, as opposed to node-centric, and work in the presence of node mobility. Extensive simulation results have shown the effectiveness of our algorithms when compared to other clustering schemes proposed in the literature.Lan Wang received his B.S. and M.S. degrees in computer science from Harbin Engineering University, China in 1992 and 1995, respectively. From 1995 to 1999, he worked as a software engineer at the System Engineering Research Institute of CSSC, Beijing, China. He is currently a PhD student at the Computer Science Department of Old Dominion University.Stephan Olariu received the M.Sc. and Ph.D. degrees in computer science from McGill University, Montreal, Canada in 1983 and 1986, respectively. In 1986 he joined the Computer Science Department at Old Dominion University where he is now a full professor. He has published extensively in various journals, book chapters, and conference proceedings. His research interests include wireless networks and mobile computing, parallel and distributed systems, performance evaluation, and medical image processing. Prof. Olariu serves on the editorial board of several archival journals including IEEE Transactions on Parallel and Distributed Systems, Journal of Parallel and Distributed Computing, International Journal of Foundations of Computer Science, Journal of Supercomputing, International Journal of Computer Mathematics, VLSI Design, and Parallel Algorithms and Applications.     

Localization of luteinizing hormone β-mRNA by in situ hybridization in the sheep pars tuberalis

Summary The localization of luteinizing hormone beta (LH)-mRNA was studied by in situ hybridization in the pars tuberalis of sheep using a homologous sheep double-stranded ³²P-or ³⁵S-cDNA. The labelled cDNA probe detected one mRNA sequence in the pars tuberalis by Northern blot analysis; this sequence was similar to that detected in the pituitary. In situ, the labelling of LH-mRNA in the horizontal and sagittal tissue sections was found throughout the pars tuberalis. This labelling was prevented by adding an excess of cold probe or treating the sections by ribonuclease before in situ hybridization. Controls showed a labelling in the pars distalis, but not in the median eminence, hypothalamus, cerebral cortex and liver sections. Double labelling by using a specific LH-antiserum indicated that the labelling of LH-mRNA appeared more intense in LH-containing cells that were found only in the ventral part of the pars tuberalis. These results suggest that the entire pars tuberalis is able to produce the LH subunit, but that the level of translation greatly varies according to the location of the cells.     

Verteilung und Aktivität hydrolytischer Enzyme in Hypophysenadenomen

Zusammenfassung An 17 Hypophysenadenomen [12 Chromophobe (endokrin-inaktive), 5 Mischtypadenome (endokrin-aktive)] wurde das Verhalten und die Aktivität der hydrolytischen Enzyme -Galaktosidase, -Glucuronidase, -Glykosidase und Arylsulfatase mit histochemischen Methoden geprüft.Die Mischtypadenome zeigen insgesamt eine höhere Aktivität als die Chromophoben. Dabei reagieren -Galaktosidase und -Glykosidase am stärksten, -Glucuronidase etwas schwächer, während Arylsulfatase die niedrigste Aktivität zeigt. Die Befunde werden mit anderen enzymhistochemischen Untersuchungen an Hypophysenadenomen und tierexperimentellen Ergebnissen verglichen. Daraus folgt, daß wahrscheinlich zwischen der Aktivität der untersuchten lysosomalen Enzyme und der endokrinen Aktivität ein Zusammenhang besteht.

---

Summary Behaviour and activity of the hydrolytic enzymes -galactosidase, -glucuronidase, -glycosidase and arylsulphatase are tested in 17 adenomas of the hypophysis (12 chromophobic, endocrine-inactive; 5 mixed cell adenomas, endocrine-active).Mixed cell adenomas show an altogether higher activity than chromophobic adenomas. -galactosidase and -glycosidase show the highest, -glucuronidase a slightly lower, and arylsulphatase the lowest activity. The findings are compared with other enzymhistochemical methods and results from animal experiments. The results of this comparison indicate that there is a correlation between the endocrine activity of the lysosomal enzymes in question.

---

---

Der Deutschen Forschungsgemeinschaft danken wir für ihre Unterstützung.

---

Fräulein Renate Kott, Fräulein Marianne Lehnen und Fräulein Edith Klasmeier danke ich für ihre technische Unterstützung.     

Activation of protein kinase C alpha is required for TPA-triggered ERK (MAPK) signaling and growth inhibition of human hepatoma cell HepG2

The signaling mechanisms for most of the antiproliferative processes are not fully understood. We have demonstrated that ERK(MAPK) signaling was involved in the induction of both p15^INK4band p16^INK4a CDK inhibitors and growth inhibition of hepatoma cell HepG2 triggered by the tumor promoter tetradecanoyl phorbol acetate (TPA). In this study, the upstream signal mechanism for TPA-induced ERK(MAPK) activation was investigated. In HepG2 cells only one of the cPKC isozymes, PKC, but not cPKCII, nPKC or aPKC was activated by TPA as demonstrated by its membrane translocation within 10–30 min and down-regulation at 24 h after TPA treatment. Pretreatment of 0.2–2.0 M Bisindolylmaleimides, an inhibitor of PKC, attenuated the TPA-induced phosphorylation of ERK, gene expressions of p15^INK4band p16^INK4a, and growth inhibition of HepG2 cell in a dose-dependent manner. Consistently, transfection of HepG2 with 1.0–3.0 M antisense (AS) PKC, but not (AS) PKCII, or nPKC oligonucleotides (ODN), for 36 h prior to TPA treatment also prevented the TPA-induced molecular and cellular effects described above. Taken together, we concluded that PKC is specifically required for TPA-induced ERK(MAPK) signaling to trigger gene expressions of p15^INK4band p16^INK4a leading to HepG2 growth inhibition.     

Structural and Functional Rearrangements of Mesophyll as a Probable Basis for the Cytokinin-Dependent Assimilate Translocation in Detached Leaves

The effects of benzyladenine (BA) on the mesophyll functioning, such as osmotic potential (), the effect of the inhibitors of ⁺-ATPase on the influx of ¹⁴C-sucrose, the direction of carbon metabolism, and the rate of dark respiration, were followed in the detached leaves of pumpkin (Cucurbita pepo L.) and broad beans (Vicia faba L.). BA elevated and established a gradient of (_p) between the treated and untreated leaf regions. The inhibitors of H⁺-ATPase did not affect the BA-induced influx of ¹⁴C-sucrose. The changes were accompanied with the elevated synthesis of starch and other polymeric compounds and the diminished synthesis of the substances of relatively low molecular weight. The stimulation of dark respiration was short and inconsiderable. The author concludes that the BA-induced transport was a passive process related to a increase. Leaf expansion accompanied by the synthesis of high-molecular-weight substances essential for cell growth and by starch synthesis apparently increased the sink capacity of the BA-treated detached leaves. The diminished efflux from the leaf blade was probably related to a lowered level of the transportable carbon compounds restricting their entry into the phloem. The influx induction could result from the activation of growth and metabolic processes, the decline in the number of organic molecules per cell volume unit, and the development of _p between the source and sink leaf regions.     

Lipochemistry of the progamic stage of a self-incompatible species: Neutral lipids and fatty acids of the secretory stigma during its glandular activity,and of the solid style,the ovary and the anther in Forsythia intermedia Zab. (Heterostylic species)

Chromatographic (thin-layer, gas column, column chromatography) analyses of neutral lipids and fatty acids of reproductive tissues of Forsythia intermedia Zab., a self-incompatible species, were performed with two objectives in mind: 1. To determine whether there is a qualitative evolution of the different classes of lipids and fatty acids that could be correlated with the three functional stages observed during previous histochemical and ultrastructural studies. The stigmatic exudate and intracellular accumulations consist mainly of neutral lipids. 2. To compare the lipid composition of the stigma (both thrum and pin forms) with that of the style, the ovary, and the anther, and to investigate the possible existence of a stigma-specific lipid compound. Stigmatic neutral lipids are found mostly in a glyceridic mixture probably containing hydrocarbons and terpenes. The fatty acids identified are between C:7 and C: 12, with the maximum unsaturated form being a C: 18. During the secretory process there is no great qualitative diference between the neutral lipids and fatty acids found in the stigmas of thrum and pin forms. Sterols are present in styles, ovaries, and anthers, but not in stigmas. They represent the only difference in the lipid composition of these various floral structures.     

Genetic control of β-diketone and hydroxy-β-diketone synthesis in epicuticular waxes of barley

Summary Five eceriferum, (cer) mutants in barley which influence -diketone and hydroxy--diketone synthesis in spike and internode epicuticular waxes have been characterized. The mutation cer-u ⁶⁹ blocks the synthesis of hydroxy--diketones and leads to a compensatory increase in the amount of -diketones, indicating that -diketones are precursors of the hydroxy--diketones. Furthermore, highly lobed wax plates were observed for the first time on barley lemmas, in addition to the characteristic wax tubes. Both diketone classes are selectively and proportionally reduced in the spike wax of cer-i ¹⁶, which has shorter wax tubes. The three mutants cer-c ³⁶, -q ⁴², and -c,u ¹⁰⁸ synthesize neither diketone class and form no wax tubes. In contrast to the variable composition of most individual barley wax classes, only a single -diketone was identified, namely hentriacontan-14,16-dione.     

Association of β<Subscript>2</Subscript>-microglobulin with the α<Subscript>3</Subscript> domain of H-2D<Superscript>b</Superscript> heavy chain

MHC class I molecules are heterotrimeric complexes composed of heavy chain, ₂-microglobulin (₂m) and short peptide. This trimeric complex is generated in the endoplasmic reticulum (ER), where a peptide loading complex (PLC) facilitates transport from the cytosol and binding of the peptide to the preassembled ER resident heavy chain/₂m dimers. Association of mouse MHC class I heavy chain with ₂m is characterized by allelic differences in the number and/or positions of amino acid interactions. It is unclear, however, whether all alleles follow common binding patterns with minimal contributions by allele-specific contacts, or whether essential contacts with ₂m are different for each allele. While searching for the PLC binding site in the ₃ domain of the mouse MHC class I molecule H-2D^b, we unexpectedly discovered a site critical for binding mouse, but not human, ₂m. Interestingly, amino acids in the corresponding region of another MHC class I heavy chain allele do not make contacts with the mouse ₂m. Thus, there are allelic differences in the modes of binding of ₂m to the heavy chain of MHC class I.     

Stimulation of membrane-associated polysaccharide synthetases by a membrane potential in developing cotton fibers

Conditions which induce a transmembrane electrical potential, positive with respect to the inside of membrane vesicles, result in a substantial (4–12-fold) stimulation of the activity of membrane-associated -glucan synthetases in a membrane preparation derived from the developing cotton (Gossypium hirsutum L.) fiber. Induction of electrical potentials which are negative with respect to the inside of the membrane vesicle results in little or no stimulation of -glucan synthesis. Those products whose synthesis is stimulated are mainly -1,3-glucan, but there is also a considerable increase in -1,4-glucan. No -1,4-glucan (starch) was detected in the reaction products. A transmembrane pH gradient was found to have no effect on -glucan synthesis. The results indicate that a transmembrane electrical potential can influence, either directly or indirectly, the activity of membrane-associated polysaccharide synthetases.Abbreviations UDP-glucose uridine-5-diphosphoglucose - PEG polyethylene glycol - BTP bistrispropane (1,3-bis[tris(hydroxymethyl)methylamino]propane) - MES 2(N-morpholino)ethanesulfonic acid - VAL valinomycin     

Expression of a chimeric stilbene synthase gene in transgenic wheat lines

A chimeric stilbene synthase (sts)gene was transferred into wheat. Stilbene synthases play a role in the defence against fungal diseases in some plant species (e.g. groundnut or grapevine) by producing stilbenetype phytoalexins like resveratrol. Resveratrol is also claimed to have positive effects to human health. Embryogenic scutellar calli derived from immature embryos of the two commercial German spring wheat cultivars Combi and Hanno were used as target tissue for cotransformation by microprojectile delivery. The selectable marker/reporter gene constructs contained the bargene either driven by the ubiquitinpromoter from maize (pAHC 25, also containing the uidAgene driven by the ubiquitinpromoter), or by the actinpromoter (pDM 302) from rice. The cotransferred plasmid pStil 2 consisted of a grapevine stscoding region driven by the ubiquitin promoter. Eight transgenic Combi and one Hanno T_Oplant were obtained and, except one Combi T_Oplant, found to be cotransformants due to the integration of both the stsgene and the selectable marker or reporter genes. Expression of the stsgene was proven by RTPCR, and, for the first time, by detection of the stilbene synthase product resveratrol by HPLC and mass spectrometry. The stsgene was expressed in four of the seven transgenic Combi T_{_o}plants. Two of the respective T₁progenies segregated in a Mendelian manner were still expressing the gene. Investigations into methylation of the stsgene showed that in three nonexpressing progenies inactivation was paralleled by methylation.     

β-Lactamases from Actinopolyspora halophila,an extremely halophilic actinomycete

Actinopolyspora halophila, an extremely halophilic actinomycete, produced both cellular and exocellular -lactamases when grown in liquid media containing 20% (w/v) NaCl. Maximal exocellular -lactamase production occurred at 48 h growth and exceeded cellular enzyme levels 7-fold. Maximal cellular -lactamase was observed at 72 h as cultures achieved full growth.Both -lactamases were purified to molecular homogeneity by a sequence involving gel filtration on Bio-Gel P-100, DEAE Bio-Gel A chromatography, preparative isoelectric focusing, and gel filtration on Sephadex G-75. Cellular -lactamase was purified 99-fold with 12% recovery and had a molecular weight of 42,200, and an isoelectric point of 4.15. Exocellular -lactamase was 12-fold purified with 1.2% recovery of initial activity and had a molecular weight of 38,000 and an isoelectric point of 3.85. Its specific activity was 7-fold greater than that of the cellular enzyme.A variety of penicillin and cephalosporin substrates were degraded by both enzymes. While the cellular -lactamase degraded phenoxymethylpenicillin, methicillin, and cephaloglycin most efficiently, the exocellular enzyme was most active against methicillin, carbenicillin, ampicillin, and cephalosporin C. Both enzymes were stimulated and protected from thermal deactivation by NaCl but not KCl or MgCl₂-Neither enzyme was inhibited by iodine.Abbreviations IEF Isoelectric focusing - SDS-PAGE sodium dodecylsulphate-polyacrylamide electrophoresis - PEG polyethyleneglycol - DTT dithiothreitol - DEAE diethylaminoethyl Issued as NRCC 25164