Enzyme Protein, 1993, 47(2), 57 - 64 A novel ornithine transcarbamylase present in mycoplasma-infected myeloma cells; Matsuno F et al.; A myeloma cell line (KHM-4) from a patient with multiple myeloma and idiopathic hyperammonemia, and another myeloma cell line (RPMI8226) were seen to have activity to form ammonia from arginine . High activity of ornithine transcarbamylase (OTC), a hepatic urea cycle enzyme, was detected in these cell lines . OTC of these cells was much more heat-stable than the liver enzyme, and did not cross-react with an antibody against the liver enzyme . As the OTC activity was also detected in the culture medium of the myeloma cells and because the activity was markedly decreased by the antimycoplasma drug MC-210, the OTC activity was assumed to be associated with mycoplasma infection . Polymerase chain reaction, using degenerate oligonucleotide mixtures corresponding to the two highly conserved sequences of OTC, amplified a DNA sequence that apparently encodes a portion (about 67% in length) of mycoplasma OTC . The predicted amino acid sequence of the mycoplasma enzyme was 33-47% identical with those of the enzymes of bacteria, yeast and mammals. Folia Med Cracov, 1993, 34(1-4), 105 - 19 Clinical ecology and its role in diagnosis of chronic diseases caused by environmental pollution . Indoor air pollution as a major factor; Krop JJ; The author discusses the four basic principles of clinical ecology, which include total load, such as exposure to chemicals, effects of physical factors (cold, heat, positive ions, radiation), exposure to bacteria, viruses and fungi, effects of consumed foods and psychological stress . Adaptation is discussed in detail as it pertains to chronic degenerative disease and chemical sensitivity . Lack of understanding of adaptive phenomena leads contemporary physicians to prescribe drugs instead of finding and eliminating the cause of disease to alleviate suffering . Body response to environmental stimuli during adaptive processes is biphasic in nature and is characterized by stimulatory and withdrawal reactions . Symptoms developed during these phases are described in detail . Effects of foods and chemicals on patients' health should be examined and considered before excluding in favour of idiopathic, mental or psychosomatic disturbances . In order to understand the variety of symptoms and signs developed during the process of degenerative disease one has to accept and apply the concept of biological individuality . Each patient should be assessed and dealt with carefully since each represents a unique clinical picture . The author further on discusses indoor air pollution, its causes and effects on health . Particular emphasis is placed on the indoor use of pesticides in the Canadian setting . Four groups of population have been defined as the result of exposure to chemicals and are discussed in brief . The author concludes with a comparison of traditional and environmental medicine in respect to history taking, the stage at which disease is diagnosed, the patient's viewpoint, specialization and therapies. Cell Mol Biol Res, 1993, 39(5), 463 - 82 The 17 kDa HBx protein encoded by hepatitis B virus interacts with the activation domains of Oct-1, and functions as a coactivator in the activation and repression of a human U6 promoter; Antunovic J et al.; The hepatitis B virus 17 kDa x gene product expressed in bacteria transactivates a human U6 promoter three- to eightfold in an ATP-independent manner in HeLa cell NTP-depleted extracts containing preassembled transcription preinitiation complexes . However, if added prior to assembly, HBx squelches the promoter . Both the HBx dependent "squelching" of U6 transcription observed in transient transfection analysis, and the transactivation observed in vitro is dependent on the presence of an upstream octamer element . HBx is incorporated via protein-protein interactions into DNA complexes containing the activation domains of Oct-1, and into a stable U6 preinitiation complex . This is consistent with a role as a coactivator interacting with the basal transcription machinery . We propose that the HBx dependent transactivation and repression of U6 transcription occurs by changes in the transcription factor limiting initiation, and propose that HBx may have a dual role in the regulation of transcription in vivo. Ann Biol Clin (Paris), 1993, 51(6), 641 - 8 {Evaluation of a new rapid ELISA technique for detecting human cytomegalovirus antibodies (CMV) . Applications in emergency blood transfusion and in organ transplantation}; Goldschmidt P et al.; Immunocubes can detect very low HCMV IgG levels with at least the same sensitivity as the reference technique: ELISA . Samples containing bacteria showed no particular interference on the test, and results for non-infected material were in agreement with those of ELISA . The rapid test did not give any false positive reactions even with samples from patients with very high antiherpes immunoglobulins levels . Samples from transplant patients at the beginning of seroconversion or diluted to IgG titers corresponding to ELISA cut-off values were always detected as positive by immunocubes . In view of the rapidity of the test, its high specificity, its ability to detect levels corresponding to extremely low IgG signals in ELISA, and the possibility of conserving objective proof of the test, we conclude that this rapid immunocube technique is of great interest for current serologic screening in an emergency context, especially for transfusion and organ transplantation. Ann Chir, 1993, 47(10), 1039 - 42 {"Pouchitis": new routes of research and signification}; Kelly KA; "Pouchitis" is clearly disease-related, in that patients with ulcerative colitis are more likely to get the condition after the ileal pouch-anal canal operation than are patients with familial adenomatous polyposis . Moreover, extraintestinal manifestations of colitis may flare up with the onset of pouchitis in colitis patients, and RFD9+ macrophages characteristics of colitis patients are more common in the pouch mucosa of patients with pouchitis than in those without pouchitis . Nonetheless, luminal factors have a role . Some, like bacteria and their products and nonsulfated bile acids, likely aggravate pouchitis, while others, like glutamine, short-chain fatty acids and perhaps calcium, seem to improve symptoms and suppress pouchitis . The inflammatory response may also be suppressed by corticoids, 5-aminosalicylic acid, allopurinol, and the immunosuppressive agents, cyclosporine and FK 506 . Nonetheless, the prevention of pouchitis is empiric and imperfect, and the response to treatment of often temporary . The exact cause and the specific cure of pouchitis are still unknown. Ann Chir, 1993, 47(10), 1029 - 33 {"Pouchitis"; histology}; Keighley MR; "Pouchitis" is a well recognised complication of restorative proctocolectomy characterised by acute diarrhoea, sometimes with blood, often complicated by incontinence, malaise, arthritis, erythema nodosum and fever . The ileal mucosa is hyperaemic, there may be shallow ulcers and contact bleeding . Biopsies characteristically show villous atrophy, a poly-morphonuclear infiltrate and chronic inflammatory cells, but the histopathological features are often patchy and may be difficult to differentiate from ischaemic ileitis, colonic metaplasia or Crohn's disease . "Pouchitis" is probably due to an overgrowth of intestinal bacteria secondary to stasis but there is some evidence that there may be an ischaemic factor . Most patients respond rapidly to oral metronidazole. Biosystems, 1993, 31(2-3), 99 - 109 Classification of organisms: living and fossil; Anderson C; The classification advanced herein provides a framework to which any organism can be readily placed and offers comprehensive coverage of the entire biological spectrum . The viruses are incorporated as a kingdom of organisms and a rationale advanced for a 6-kingdom system . Many seldom-classified fossil organisms are included, with a significant number of them classified at an advanced level . Individual phyla are recognized for the fossil genera Tribrachidium, Amiskwia, Dinomischus, Anomalocaris, Tullimonstrum, Hallucigenia, Opabinia and Pikaia . Other seldom-recognized phyla include viruses, conularids, loriciferans, tentaculites, hyoliths, scleritophores, myzostomans, acorn worms, conodonts, monoblastozoans and volborthellids . The notion that failures (geologically) cannot be phyla is rejected . A system of kingdom-related and rank-related suffixes is incorporated with the flexible opportunity, if deemed necessary, to advance or reduce taxa by re-suffixing . Other features of the complete classification of Anderson (Classification of Organisms: Living and Fossil, Golden Crowns Press, Lancaster, OH, 1992) include the elimination of duplicate taxa, incorporation of geologic ranges and number of species for many groups, and extensive usage of common names, which are completely indexed. Clin Rev Allergy, 1993 Winter, 11(4), 471 - 90 The role of allergens in atopic dermatitis; Jones SM et al.; AD is a disorder that affects up to 12% of the pediatric population . This disease is multifactorial and encompasses a wide array of etiologic factors . Strong evidence has existed in the literature over the past century for the role of inhalant and food allergies in the pathogenesis of AD . Much work is currently ongoing to delineate the role of individual cellular components, cytokines, and other mediators in the pathogenesis of AD . The answers to these questions, as well as a more comprehensive understanding of hereditary factors, will provide key information to our overall understanding of AD and our ability to treat patients with this disease more effectively in the future. Rev Elev Med Vet Pays Trop, 1993, 46(1-2), 309 - 11 The epidemiology and control of camel dermatophilosis; Gitao CG; Camel dermatophilosis was only recently described . It appears however that it is more widespread than originally thought . In Kenya it has generally been found in the main semi-arid camel rearing areas of Samburu and Laikipia districts although it has not yet been found in the arid areas of Turkana district . In an investigation of ticks on 200 camels, no Amblyomma variegatum ticks were found although many other ticks were present . A . variegatum is suspected to transmit dermatophilosis in many domestic animals . The only control method of dermatophilosis currently practised in Kenya is in one commercial farm, where camels are regularly washed with a 1% potassium aluminium sulphate solution . The camels have shown progressive improvement . Recently, some 50 camels imported from Pakistan in this farm came down with a severe skin infection which closely resembled dermatophilosis . All imported adult camels were involved although no calves were involved . Since no bacteria were isolated from all the sick camels, it was thought to be due to vitamin deficiency. Probl Tuberk, 1993, (4), 51 - 2 {Production of purified antigenic drug from Mycobacterium tuberculosis humanus and its use for serodiagnosis of tuberculosis}; Baenskii AV et al.; A variety of antigenic preparations from whole bacteria M . tuberculosis humanus (H37Rv) and relevant subcellular components have been tested using indirect ELISA for determination of antituberculous antibodies . Cell membranes of mycobacteria and antigens isolated from KCl-extracts of cell membranes in gel Toyopearl HW 55 F demonstrated the highest activity . Fraction 6 usage raised the reaction sensitivity to 83.3% in specificity 86.7% . As indicated by the ELISA, the highest antibodies content under the above antigen introduction was reached in patients with fibrocavernous tuberculosis, the lowest one in focal tuberculosis. Vet Res, 1993, 24(6), 503 - 14 {Microclimate and air composition in a closed chamber meant for the study of the toxicity of atmospheric pollutants for the piglet}; Urbain B et al.; An experimental model representing the pollution background in a pigpen was constructed in order to study the toxicology of pig respiratory air pollutants . Temperature, relative humidity, air-flow rate, total dust, total viable particles, endotoxins in respirable dust (< 5 microns) and ammonia were measured in a 1.9 m3 environmental chamber designed for piglets . The activity of 1 piglet was recorded on video . Temperature and relative humidity were respectively 23.9 +/- 1.3 degrees C and 70.5 +/- 8.3% (mean +/- SD) . Air flow rate was 10 m3/h . Dust, viable particles, endotoxin and ammonia concentrations were respectively 7 +/- 2 particles/ml, 3.4 x 10(4) +/- 2.9 x 10(4) BCFP (bacterial colony-forming particles)/m3, 4.8 +/- 1.5 ng/m3 and 6 +/- 0.8 ppm (mean +/- SD) . All these parameters were comparable to the lowest values recorded in pigpens. Mol Microbiol, 1993 Jan, 7(2), 285 - 8 Q pili enhance the attachment of Moraxella bovis to bovine corneas in vitro; Ruehl WW et al.; Moraxella bovis, the causative agent of infectious bovine keratoconjunctivitis, exhibits several virulence factors, including pili, haemolysin, leukotoxin, and proteases . The pili are filamentous appendages which mediate bacterial adherence . Prior studies have shown that Q-piliated M . bovis Epp63 are more infectious and more pathogenic than I-piliated and non-piliated isogenic variants, suggesting that Q pili per se, or traits associated with Q-pilin expression, promote the early association of Q-piliated bacteria with bovine corneal tissue . In order to better evaluate the role of Q pili in M . bovis attachment, several M . bovis strains and a recombinant P . aeruginosa strain which elaborates M . bovis Q pili but not P . aeruginosa PAK pili, were evaluated using an in vitro corneal attachment assay . For each strain tested, piliated organisms attached better than non-piliated bacteria . M . bovis Epp63 Q-piliated bacteria adhered better than either the I-piliated or non-piliated isogenic variants . Finally, recombinant P . aeruginosa organisms elaborating M . bovis Q pili adhered better than the parent P . aeruginosa strain which did not produce M . bovis pili . These results indicate that the presence of pili, especially Q pili, enhances the attachment of bacteria to bovine cornea in vitro. Biull Eksp Biol Med, 1993 Jan, 115(1), 77 - 9 {Electron-autoradiographic study of burn wound healing while treated with levosin ointment}; Chekmareva IA et al.; Electron-microscopic radioautography was employed to study the crust and granulation tissue from patients with thermal burn injuries degree II-IV, before treatment, 10 and 20 days after application of levosin ointment . Local introduction of the ointment promoted the burn healing as shown by an increase in the number of fibroblasts and vessels of the granulation tissue, by activation of fibroblast function and proliferation, by destruction of bacteria and reduction in their number. Vet Res Commun, 1993, 17(6), 429 - 47 Natural killer (NK) cells in domestic animals: phenotype, target cell specificity and cytokine regulation; Evans DL et al.; A comprehensive review of NK cells in animals of veterinary medical importance has not been previously published . However, these cells have a high level of immunological/medical relevance due to their role in tumour cell destruction and B-cell regulation, as well as their inhibitory activities against various parasites and bacteria . In the present review, NK cells from agriculturally important animals are characterized . Cell phenotype descriptions have shown that for each species, unique (i.e . non-cross-reactive with anti-human CD antibodies) and different monoclonals are required to identify NK cells . These cells lyse certain tumour and virus transformed target cells and, as might be expected from the diverse species compared in this review, analysis of the tissue distribution of NK cells gives highly varied results . NK cell morphology differs in these species from small agranular to large granular lymphocytes . The final area considered relates to studies on the effects of cytokines on NK function and to research identifying which cytokines (if any) are produced by NK cells during activation responses . The largest quantity of available data concerns functional and descriptive studies in these animals . However, it is appropriate to consider this research as a starting point on which to build comparative and molecular studies of the roles of NK cells in immunosurveillance and immunoregulation. Rocz Panstw Zakl Hig, 1993, 44(2-3), 171 - 3 {Tests of Helicobacter pylori survival in milk}; Kafel S et al.; The survival of microaerophilic bacteria Helicobacter pylori (Hp) isolated from gastric mucosa was studied in 2 ml and 10 ml samples of milk contaminated with a suspension of these bacteria in 20% glucose solution . Milk was kept at 5-7 degrees C temperature in refrigerator and at room temperature . Hp was found in samples kept at 5-7 degrees C during 3 days . On the other hand, Hp was not found after 24-hours keeping at room temperature. Rev Chil Obstet Ginecol, 1993, 58(5), 349 - 54 {Mycoplasma hominis and Ureaplasma urealyticum in pregnant and infertile women . Differences in tubal pathology}; Reid I et al.; We studied the frequency of cervicovaginal bacteria in 47 pregnant women (16-41 weeks of gestational age), in 47 infertile patients and in 4 with gynecological diseases (not infections) . When we compared the frequency of bacteria between pregnant and infertile women, we found that the biggest difference was with U . urealyticum . In fact the frequency of this germ was 61% in the total population studied, in pregnant women was of 72% which is statistically higher than the frequency in infertile women which was of 47% (p < 0.025) . We think that the reason for the higher frequency of this germ is a consequence of the effect of estrogens that favors the proliferation of this of germ . In the case of M . hominis the frequency in pregnant women was of 13% and in infertile women was of 21% which was not statistically different . Within the group of infertile patients, those that had tubal damage had a higher frequency of germs that the patients that did not have damage, moreover M . Hominis was not detected in the infertile patients without tubal damage and in patients with tubal damage its frequency was of 25%. Eisei Shikenjo Hokoku, 1993, (111), 149 - 50 {Significance of mutagenicity tests on safety evaluation of medicinal drugs}; Sofuni T; As one of toxicological tests for safety evaluation of medicinal drugs, three representative mutagenicity tests, that is, reverse mutation test in bacteria, chromosomal aberration test with mammalian cells in culture and micronucleus test with rodents have been carried out . These three tests, especially two in vitro tests have been used for initial hazard identification . Further, the micronucleus test using experimental animals has offered significant information for in vivo safety evaluation . Based on recent collaborative studies with mouse micronucleus tests on human carcinogens and results from newly developed in vivo gene mutation assay using transgenic mice, the combination of both test systems seems to be powerful for in vivo safety evaluation. Neuropsychobiology, 1993, 28(1-2), 110 - 2 Direct innervation of lymphoid organs: substrate for neurotransmitter signaling of cells of the immune system; Felten DL; The nervous system can generate outflow capable of signaling cells of the immune system via two routes: (1) hormonal influences via hypothalamopituitary-target organ activation, and (2) neurotransmitter influences via direct innervation of the parenchyma of both primary and secondary lymphoid organs . Both noradrenergic and neuropeptidergic nerve fibers are found directly adjacent to cells of the immune system in bone marrow, thymus, spleen, lymph nodes, and other sites . These neurotransmitters can modulate immune responsiveness at several levels, including individual cellular functions, collective cellular interactions and immune responses, and overall host immunological response to a challenge by bacteria, viruses, tumor cells, or other sources . The ability of neural signals to modulate immune responsiveness is influenced by such factors as co-localization with other neurotransmitters, synergistic or dual signaling by neurotransmitters and cytokines, hormonal influences on ligand availability or receptor responsiveness, extent of catabolism and inactivation of the neural signal, and unique chemical and morphological aspects of the specific compartmentation of the lymphoid organ in which signaling is taking place . Thus, neural-immune signaling is a complex and dynamic process, with bidirectional interactions in a complex microenvironment. Bull Soc Pathol Exot, 1993, 86(5 Pt 2), 489 - 91 {Javelle water and drinking water}; Jacquet F; Because of its exceptional disinfecting properties, hypochlorite bleach is, along with vaccines and antibiotics, an outstanding tool in the prevention of water-borne infectious diseases such as cholera, and especially in tropical environments . In addition, hypochlorite bleach is available around the world, and at low cost . Moreover, hypochlorite bleach is safe for human when used as directed . It is also sale for the environment in household use . In the Public Health domain, hypochlorite bleach with its 200-year history remains both a modern product and a product of the future. Biotechnology (N Y), 1993 Jan, 11(1), 60 - 3 Application of magnetic beads in bioassays; Haukanes BI et al.; This review outlines the possibilities of magnetic separation techniques and the application of magnetic beads in bioassays . By linking monoclonal antibodies or DNA to magnetic beads, or by using magnetic beads coated with streptavidin, a specific interaction with the corresponding target is ensured . By means of an external magnet, the recovery of material for further studies is greatly simplified . Magnetic beads have proven valuable in cell separations, for example, removal of tumor cells from bone marrow and isolation of lymphoid cells from peripheral blood, and for the isolation, identification and genetic analysis of specific nucleic acid sequences (DNA or RNA) and for isolation of DNA binding proteins . In addition, some of these techniques have also proven to be useful in the detection of specific nucleic acids from viruses or bacteria. Microbiol Immunol, 1993, 37(7), 563 - 71 Evaluation of opsonophagocytic dysfunctions in severely burned patients by luminol-dependent chemiluminescence; Ono Y et al.; We compared the luminol-dependent chemiluminescence (CL) response of peripheral blood from severely burned patients with that from normal controls to evaluate the primary defense level against bacterial infection in the patients . The CL was measured upon addition to diluted whole blood of a soluble stimulus, phorbol myristate acetate (PMA) or particulate stimuli such as bacteria or zymosan without special opsonization . In the early post-burn days, the initial rate of whole blood CL induced with the particulate stimulus was much lower than that in the normal controls, whereas the rate was higher when PMA was used as a stimulus . The number of granulocytes in the patients' blood had increased and isolated polymorphonuclear leukocytes (PMNs) from the patients exhibited higher CL responses to the particulate or soluble stimulus as compared with those of normal controls . The results suggest that the PMNs in burn patients were activated and normally mobilized in the early post-burn period but the opsonizing capacity in the blood decreased . In fact, the serum levels of complement, immunoglobulins and fibronectin were found to be lower in the blood from the patients than those from normal controls and a supplement of freshly frozen plasma of human immunoglobulin preparations restored the initial rate of the whole blood CL upon phagocytosis . The prognosis is still poor when severe infection occurs in the patients with decreased CL response of whole blood.Recombinant human granulocyte colony-stimulating factor (rh G-CSF) enhanced the CL response of PMNs from burn patients . The administration of rhG-CSF may be useful for decreasing the morbidity of severe infection following burn injury in the near future. Methods Enzymol, 1993, 225, 587 - 610 Construction of primary and subtracted cDNA libraries from early embryos; Rothstein JL et al.; By modifying current cDNA cloning and electroporation methods, large and representative murine cDNA libraries were synthesized from 10 to 100 ng mRNA isolated from unfertilized egg and preimplantation mouse embryos . High cloning efficiency is essential for complete representation of genes expressed in egg and preimplantation embryos and for the isolation of stage-specific genes using subtractive hybridization . Because the mouse embryo contains no more than 50 pg of poly(A)+ mRNA at any stage of preimplantation development, approximately 5000-10,000 embryos are required to obtain enough mRNA to synthesize libraries using current methods . To obtain a representative library that also includes rare transcripts, the size of the library should be at least 10(6) clones . The average percent conversion of mRNA to single-stranded cDNA was 20-40%, so that a cloning efficiency of nearly 2 x 10(8) cfu/microgram cDNA is required for such a cDNA library . No previous methods have provided directional cloning of cDNA into plasmids with these high efficiencies . The advent of electroporation methods for the introduction of nucleic acids into bacteria has made possible the use of standard plasmid vectors for high-efficiency cDNA cloning . Plasmid vectors are currently available that can accommodate the directional cloning of cDNA such that T7 and T3 RNA polymerase promoter sequences can be used to generate sense and anti-sense transcripts for subtractive hybridization and riboprobe synthesis . The cDNA libraries we derived using this methodology are a reusable and abundant source of genetic information about the control of preimplantation development . Specialized subtractive cDNA libraries enriched for genes expressed exclusively at a predetermined time in development give access to genes expressed in a stage-specific manner . The ability to construct new cDNA libraries from limited amounts of starting material ensures the provision of new and important resources for the identification and study of novel genes or gene families, and it is an important new tool for understanding the molecular control of mammalian development. Clin Lab Haematol, 1993, 15(2), 79 - 85 Pilot study: effects of G-CSF on neutrophil ex-vivo function post bone marrow transplantation; Macey MG et al.; The effects of human recombinant granulocyte colony stimulating factor (G-CSF) on neutrophil recovery have been studied in patients undergoing bone marrow transplantation for haematological disease . Nine patients (five autografts and four allografts) were studied while receiving daily doses of G-CSF (range 2-20 micrograms/kg body weight) were compared to eight patients (four autografts and four allografts) who did not receive G-CSF as a control group . In both groups flow cytometry was used to determine neutrophil IgG Fc receptor (FcRII, FcRIII) expression . Phagocytosis and metabolic burst was assessed using IgG opsonized bacteria . The patients' neutrophils were studied prior to conditioning in autografts and donor cells were studied in allografts . Studies were repeated at neutrophil recovery (neutrophil count 1.0 x 10(9)/l) and ten days post recovery . At recovery FcRII expression was slightly increased in all groups, whereas the number of cells expressing FcRIII was reduced . This reduction was significant in the patients receiving rG-CSF at recovery and post recovery compared to the pretreatment levels . The ability to phagocytose bacteria in cells from patients receiving G-CSF was also reduced . The associated metabolic burst was significantly reduced in the autograft group but not the allograft group receiving rG-CSF . The reduced function and reduced expression of FCRIII suggests the presence of immature neutrophils in patients receiving rG-CSF post BMT. Crit Rev Microbiol, 1993, 19(2), 99 - 115 Biology, fiber-degradation, and enzymology of anaerobic zoosporic fungi; Wubah DA et al.; Anaerobic zoospore-producing fungi that inhabit the gastrointestinal tract of herbivores, especially ruminants, have recently been discovered . These fungi have been isolated from the rumen, hind gut, and the feces of ruminants . Thirteen species, belonging to five genera, of these fungi have been assigned to the class Chytridiomycetes . These species are classified according to the number of flagella on the zoospores and the types of thalli that develop from the zoospores . Their life cycle consists of a zoospore that encysts and develops into a vegetative thallus with zoosporangia, which at times become resting sporangia . These fungi produce a wide range of active hydrolytic enzymes, notably cellulases and xylanases, that provide them with the potential to degrade the major structural polysaccharides in plant cell walls . Their cellulases are among the most active reported to date and solubilize both amorphous and highly ordered cellulose . Their esterases are active against both feruloyl and p-coumaroyl arabinoxylans, which provides an advantage in degrading poorly biodegradable cell walls . They degrade lignin-containing cell walls, but do not metabolize the lignin moiety . Rhizoids of vegetative thalli penetrate cell walls, and they are better able than bacteria or protozoa to attack recalcitrant tissues and weaken the textural strength of plant material. Gut, 1993, 34(2 Suppl), S74 - 6 Biological basis for the clinical use of interferon; Dianzani F; Interferons are proteins produced by certain cells in response to stimuli such as foreign cells (including tumour cells), bacteria, and viral antigens . They interact both with the interferon producing cells and other cells through production of effector proteins . There are three main types of interferons, known as alpha, beta, and gamma, which have direct antiviral and immunomodulatory effects . Antiviral effects may include inhibition of viral replication, protein synthesis, maturation, or release from infected cells . Immunomodulating effects may include enhancement of macrophage, cytotoxic T cell, and natural killer cell activity . In chronic viral hepatitis, the precise mechanisms of action of alpha interferon are not yet certain . Patients with chronic hepatitis B, however, have been shown to lack endogenous interferon production; those who respond to alpha interferon treatment show a characteristic peak in serum amino-transferase activity before resolution of the infection, indicating an immune reaction . In chronic hepatitis C, the antiviral effect may be more important; patients who respond to alpha interferon tend to have higher values of 2'5' oligo adenylate synthetase, an enzyme induced by interferons that breaks down viral RNA . The clinical relevance of anti-interferon neutralising antibodies produced by some patients during interferon treatment has yet to be firmly established. Drug Alcohol Depend, 1993 Jan, 31(2), 139 - 47 A discriminant function analysis of various interferon parameters among alcoholics and heavy smokers; Windle M et al.; A three-group design of alcoholics, heavy smokers and controls was used to investigate the status of the interferon system, including natural killer cell activity . Group differences were indicated via discriminant function analysis which correctly classified 86% of subjects . Test-retest relations were investigated for 14 subjects following a 30-day alcoholic inpatient program . Whereas significant immune suppression was indicated at the time of detoxification, recovery was evident at the 30-day follow-up . Results are discussed in terms of the significance of alcohol abuse on immune system functioning with consequences for susceptibility to viruses, bacteria and medical illnesses. J Clin Periodontol, 1993 Jan, 20(1), 1 - 6 Effectiveness of open flap root debridement with rubber cups, interdental plastic tips and prophy paste . An SEM study; Schwarz JP et al.; This study was designed to ascertain whether conservative cleaning of surgically-exposed root surfaces can achieve complete plaque removal . 10 single-rooted teeth (40 surfaces) from 4 patients with advanced periodontitis were included in the study . After flap reflection, the root surfaces were cleaned using only rubber cups, EVA plastic tips and prophy paste . No attempt was made to remove calculus . Immediately after treatment, the teeth were extracted . Then root surfaces were systematically examined in the scanning electron microscope to detect any residual bacteria (plaque) . 27 of the 40 treated root surfaces were plaque-free . On the other 13 root surfaces, only a few isolated small islands of plaque were detected . On the other hand, relatively extensive areas of the root surfaces exhibited calculus . Bacterial plaque accumulation was routinely observed on the rough calculus surfaces and at the periphery of the hard deposits . These results demonstrate that the instruments used in this study can successfully remove plaque from exposed root surfaces . However, subgingival calculus that is firmly attached to root surfaces virtually always harbors plaque bacteria; such deposits require more aggressive instrumentation (scalers, curettes) for removal. Mutat Res, 1993 Jan, 285(1), 109 - 16 Directed mutation: paradigm postponed; MacPhee DG; Claims have been made in recent years that spontaneous mutational events are "directed" by (i) the presence in a selective medium of a single carbon source which the cells are unable to utilize (e.g . lactose), or (ii) the absence from a selective medium of an amino acid which the cells are unable to make for themselves (e.g . tryptophan) . In resurrecting the previously rejected hypothesis of a "directed" origin for spontaneous mutants, it was noted that selecting for valine-resistant mutants did not allow comparable increases in colony numbers to be observed when overlays containing valine and glucose were added to plates which had accumulated (or were still accumulating) large numbers of "directed" Lac+ or Trp+ mutants . The present paper shows that Valr mutants do occur in much greater numbers if a carbon source other than glucose is added to the plates along with the valine-containing overlays; the evidence is that even small amounts (< 0.02%) of glucose in the overlays prevent the recovery of Valr mutants . From these results, it is argued that the apparent "directed" nature of the spontaneous mutation process is actually a manifestation of the long-known phenomenon of glucose repression. Rev Elev Med Vet Pays Trop, 1993, 46(1-2), 257 - 61 Culture filtrate proteins of Dermatophilus congolensis; el Jack MA et al.; In previous studies on the antigens of Dermatophilus congolensis very little attention has been given to the hyphae and to excreted-secreted products (ESP) of actively growing bacteria . In this study we have grown four isolates of D . congolensis in a serum free synthetic liquid culture medium based on RPMI 1640 . Diafiltration and ultrafiltration were used to prepare ESP from infected culture fluid . These methods produced sufficient quantities of ESP that the polypeptide profiles of the four isolates could be examined by SDS-PAGE and Western immunoblotting . The four isolates produced a large number of polypeptides in culture, most of which were produced by all four isolates . However each isolate produced polypeptides that were unique to it . Western immunoblotting studies using pooled sera from chronically affected animals from Ghana showed that a number of polypeptides in ESP of a Ghanaian isolate were antigenic . When the same sera was tested against ESP from a Scottish isolate a number of polypeptides of the same molecular weight as those in the Ghanaian isolate and some at different molecular weights were recognized . This indicates that isolates of D . congolensis from different geographical areas produce ESP with shared antigenic determinants. Annu Rev Microbiol, 1993, 47, 945 - 63 The Tn5 transposon; Reznikoff WS; The bacterial transposon Tn5 encodes two proteins, the transposase and a related protein, the transposition inhibitor, whose relative abundance determines, in part, the frequency of Tn5 transposition . The synthesis of these proteins is programmed by a complex set of genetic regulatory elements . The host DNA methylation function, dam, inhibits transposase promoter recognition and indirectly enhances the transposition inhibitor promoter . The inhibitor lacks the N-terminal 55 amino acids of the transposase, suggesting that this sequence plays a key role in the transposition process . An intact N-terminal sequence is required for the transposase's recognition of the 19-bp end DNA sequences . This is the first critical step in the transposition process . Transposase-end DNA interaction is itself regulated by an intricate series of reactions involving several host proteins: DnaA, Dam, and Fis . The transposase is a unique protein in that it acts primarily in cis and inhibits its own activity in trans . Models to explain these properties are described . Finally circumstantial evidence suggests that transposition occurs preferentially from newly replicated DNA that has yet to be partitioned to progeny cells . This timing of transposition is likely to have a selective advantage for the host and the transposable element. Springer Semin Immunopathol, 1993, 15(2-3), 155 - 71 Evolution and pathophysiology of the human natural anti-alpha-galactosyl IgG (anti-Gal) antibody; Galili U; Anti-Gal is a human natural antibody which interacts specifically with the mammalian carbohydrate structure Gal alpha 1-3Gal beta 1-4GlcNAc-R, termed, the alpha-galactosyl epitope . This antibody constitutes approximately 1% of circulating IgG in human serum and is produced, upon stimulation, by 1% of circulating B lymphocytes . Anti-Gal is also present as IgA antibodies in body secretions such as saliva, milk and colostrum . The antigenic source for the constant production of anti-Gal seems to be the alpha-galactosyl-like epitopes found on many bacteria of the gastrointestinal flora . Whereas anti-Gal is abundant in humans, apes and Old World monkeys, it is absent from New World monkeys, prosimians and nonprimate mammals . The latter group of species produces, however, large amounts of alpha-galactosyl epitopes (> 10(6) epitopes per cell) . It is estimated that anti-Gal appeared in ancestral Old World primates less than 28 million years ago, possibly as a result of an evolutionary event which exerted a selective pressure for the suppression of alpha-galactosyl epitopes expression by inactivation of the gene for the enzyme alpha 1,3 galactosyltransferase . This also resulted in the loss of immune tolerance to the alpha-galactosyl epitope and the production of anti-Gal . The physiologic role of this antibody is not clear as yet . It may participate in the protection against gastrointestinal bacteria . In addition it seems to contribute to the removal of normal and pathologically senescent red cells by interacting with the few hundred cryptic alpha-galactosyl epitopes which are exposed de novo in the course of red cell aging, thereby opsonizing these cells for phagocytosis by reticuloendothelial macrophages . The alpha-galactosyl epitope has been found to be aberrantly expressed on human cells and the interaction of anti-Gal with such epitopes may result in autoimmune disease . Preliminary data suggest such a mechanism in Graves' disease . Anti-Gal has been found to interact with therapeutic recombinant proteins expressing alpha-galactosyl epitopes, but so far there is no indication that it affects the half-life in the circulation and the biologic activity . Detection of anti-Gal in the seminal fluid and in the cerebrospinal fluid may serve as a simple means for assessment of damage to the blood-genital tract barrier or the blood-brain barrier . Studies on the interaction of anti-Gal with aberrantly expressed alpha-galactosyl epitopes on human cells may elucidate the possible role of anti-Gal in human autoimmune diseases. Ann Fr Anesth Reanim, 1993, 12(3), 293 - 307 {Severe acute pancreatitis: diagnostic approaches and therapeutic implications}; Tanguy M et al.; Diagnosis of acute pancreatitis (AP) can be obtained with a high level of accuracy by clinical assessment and determination of common laboratory parameters such as serum amylase and lipase concentrations . However, the key of an optimal management of patients with AP is based on an early discrimination between interstitial oedematous and necrotizing forms . The former resolves spontaneously whereas parenchymal necrosis acting as a focus for bacteria has a very high severity . In this respect, multifactor prognostic scoring systems and new biological assessments like C reactive protein are valuable methods for forecasting the prognosis of AP . However, these indicators of severity require a full 48 hour period of observation . In order to overcome these drawbacks, other prognostic criteria have been explored based mainly, on laboratory data . The most interesting ones are trypsinogen activation peptides and leucocyte elastase . Finally, the more useful tool is computed tomography (CT) . Combined with high dose intravenous contrast agent, it allows an early identification of necrosis . Other goals of computed tomography are an accurate diagnosis of infection by guided needle aspirations and a preoperative recognition of devitalized and infected tissues, which require a careful surgical necrosectomy . A prolonged drainage is always recommended but relative merits of a conventional closed drainage and an open one are controversial . Another therapeutic challenge is gallstone associated to severe pancreatitis . An early stone removal is advocated by some authors but others prefer delayed surgery because of high mortality rates in case of emergency surgery . Delayed surgery until biological parameters of pancreatitis are normalized seems preferable . An early endoscopic sphincterotomy in an attractive alternative method. Biochem Biophys Res Commun, 1992 Dec 30, 189(3), 1296 - 302 Molecular characterization of the gene encoding glutamine synthetase in the cyanobacterium Calothrix sp . PCC 7601; Elmorjani K et al.; In order to study the regulation of the synthesis of glutamine synthetase in response to changes in environmental parameters (light and nitrogen sources), we have cloned and sequenced the glnA gene from the filamentous cyanobacterium Calothrix PCC 7601 . This gene consists of 472 codons and encodes a polypeptide of M(r) 52,290 highly homologous to that from Anabaena PCC 7120, but more distant from those identified from other procaryotes . The relative abundance of the two glnA transcripts (1.6 and 1.8 kb) is equivalent in cells grown under either red or green light, but the 1.6-kb species predominates in nitrate-grown cells and the 1.8-kb species in ammonia-grown cells . The very high identity (74%) observed between the 374-bp long nucleotide sequence upstream from the Calothrix and Anabaena glnA genes suggests the existence of similar regulatory signals for the control of glnA expression in both cyanobacteria. Cell, 1992 Dec 24, 71(7), 1073 - 80 High frequency mutagenesis by a DNA methyltransferase; Shen JC et al.; HpaII methylase (M . HpaII), an example of a DNA (cytosine-5)-methyltransferase, was found to induce directly a high frequency of C-->U transition mutations in double-stranded DNA . A mutant pSV2-neo plasmid, constructed with an inactivating T-->C transition mutation creating a CCGG site, was incubated with M . HpaII in the absence of S-adenosylmethionine (SAM) . This caused an approximately 10(4)-fold increase in the rate of reversion when the mutant neo plasmid was transformed into bacteria lacking uracil-DNA glycosylase . The mutation frequency was very sensitive to SAM concentration and was reduced to background when the concentration of the methyl donor exceeded 300 nM . The data support current models for the formation of a covalent complex between the methyltransferase and cytosine . They also suggest that the occurrence of mutational hot spots at CpG sites may not always be due to spontaneous deamination of 5-methylcytosine, but might also be initiated by enzymatic deamination of cytosine and proceed through a C-->U-->T pathway. Biochemistry, 1992 Dec 22, 31(50), 12665 - 72 Studies on Na+ and H+ translocation through the Fo part of the Na(+)-translocating F1Fo ATPase from Propionigenium modestum: discovery of a membrane potential dependent step; Kluge C et al.; The purified ATPase of Propionigenium modestum (F1Fo) was incorporated into liposomes, and the F1 part was dissociated . The Fo-liposomes catalyzed proton uptake in response to a potassium diffusion potential (inside negative) . Proton translocation was abolished by rebinding F1 to the Fo-liposomes or after incubation with the c-subunit-specific inhibitor dicyclohexylcarbodiimide (DCCD) . Proton uptake was also sensitive to the presence of external Na+ or Li+ ions and was completely abolished at 2 mM NaCl or 150 mM LiCl, respectively . However, the same concentrations of these salts in the internal volume of the Fo-liposomes were without effect, suggesting that the cation binding site is not accessible from both sides of the membrane simultaneously . An open channel-type of transport through Fo from P . modestum is therefore excluded . The Fo-liposomes also catalyzed Na+ influx or efflux in response to a K+ diffusion potential that was negative on the inside or outside, respectively . These Na+ fluxes could not be created, however, by delta pNa+ of about 60-180 mV . The initial rate of Na+ uptake depended strongly on the size of the membrane potential with no significant conductivity below -40 mV, followed by a proportional increase up to about -115 mV . In the absence of a membrane potential, the Fo-liposomes catalyzed 22Na+ counterflow against a 28-fold concentration gradient . Uptake of 22Na+ into Fo-liposomes against delta pNa+ (counterflow) was completely prevented by imposing an inside-positive potassium diffusion potential of 90 mV . The catalysis of 22Na+ counterflow by Fo from P . modestum is a clear indication of a carrier (transporter)-type mechanism and excludes a channel mechanism.(ABSTRACT TRUNCATED AT 250 WORDS) FEMS Microbiol Lett, 1992 Dec 15, 79(1-3), 361 - 70 Chromatins of low-protein content: special features of their compaction and condensation; Kellenberger E et al.; Histonic chromatin with a relatively high-protein content (RPC of about 1) is compared with naturally occurring chromatins of low-protein contents (RPCs of less than 0.5) . The features of these chromatins, with respect to compaction and condensation, are discussed . Liquid crystalline chromatin, as found in dinoflagellates and phage heads, can apparently only be formed by condensation of chromatin of low-protein content and when it is not supercoiled . With histonic chromatin, liquid crystals are never found . Chromatins with low-protein contents might also form compactosomes (or 'labile nucleosomes'), as, for instance, in bacteria . They are forms of supercoiled DNA without a protein core and are so labile that they are difficult to study and even to detect . Chemical fixatives, as commonly used for electron microscopy, do not cross-link the chromatins of low-protein content, a feature which they share with naked DNA . It is postulated that these fixatives even relax the existing supercoil, which seems to be preserved after cryofixation only. FEMS Microbiol Lett, 1992 Dec 15, 79(1-3), 273 - 80 Specific removal of chlorine from the ortho-position of halogenated benzoic acids by reductive dechlorination in anaerobic enrichment cultures; Gerritse J et al.; Anaerobic enrichment cultures catalysing the reductive dechlorination of chlorinated benzoic acids were obtained from three fresh-water sediments collected from seven different locations . Sub-cultures from these enrichments specifically removed ortho-substituted chlorine from 2,3,6-, 2,3,5- and 2,4,6-trichlorobenzoic acid, yielding chloride and 2,5-, 3,5-, and 2,4-dichlorobenzoic acids, respectively . These reductive dehalogenations were stimulated by the addition of benzoate and/or volatile organic acids . In one of these enrichments dehalogenation of ortho- and/or para-chlorine substituents was also observed from 2,3-, 2,4-, 2,5-, and 3,4-dichlorobenzoic acid, yielding 3- and 4-chlorobenzoate . Removal of meta-chlorines was not observed in any of the enrichments. Eur J Biochem, 1992 Dec 15, 210(3), 849 - 54 Purification and characterization of glycerate kinase from a serine-producing methylotroph, Hyphomicrobium methylovorum GM2; Yoshida T et al.; The glycerate kinase of a serine-producing methylotroph, Hyphomicrobium methylovorum GM2, was purified to complete homogeneity and characterized, the first time for an enzyme from a methylotroph . The enzyme was a monomer with a molecular mass about 41-52 kDa . The enzyme was stable against heating at 35 degrees C for 30 min at pH values over 6-10 . Maximum activity was observed at pH 8.0 and around 50 degrees C . The Km values for D-glycerate and ATP were 0.13 mM and 0.13 mM, respectively . The enzyme showed high specificity for D-glycerate, and was activated by potassium and ammonium ions . The reaction product of the enzyme was identified as 2-phosphoglycerate. Proc Natl Acad Sci U S A, 1992 Dec 15, 89(24), 12180 - 4 Papillomavirus L1 major capsid protein self-assembles into virus-like particles that are highly immunogenic; Kirnbauer R et al.; Infection by certain human papillomavirus types is regarded as the major risk factor in the development of cervical cancer, one of the most common cancers of women worldwide . Analysis of the immunogenic and structural features of papillomavirus virions has been hampered by the inability to efficiently propagate the viruses in cultured cells . For instance, it has not been established whether the major capsid protein L1 alone is sufficient for virus particle assembly . In addition, it is not known whether L1, L2 (the minor capsid protein), or both present the immunodominant epitopes required for induction of high-titer neutralizing antibodies . We have expressed the L1 major capsid proteins of bovine papillomavirus type 1 and human papillomavirus type 16 in insect cells via a baculovirus vector and analyzed their conformation and immunogenicity . The L1 proteins were expressed at high levels and assembled into structures that closely resembled papillomavirus virions . The self-assembled bovine papillomavirus L1, in contrast to L1 extracted from recombinant bacteria or denatured virions, also mimicked intact bovine papillomavirus virions in being able to induce high-titer neutralizing rabbit antisera . These results indicate that L1 protein has the intrinsic capacity to assemble into empty capsid-like structures whose immunogenicity is similar to infectious virions . This type of L1 preparation might be considered as a candidate for a serological test to measure antibodies to conformational virion epitopes and for a vaccine to prevent papillomavirus infection. FEMS Microbiol Lett, 1992 Dec 15, 79(1-3), 59 - 65 DNA extraction for 16S rRNA gene analysis to determine genetic diversity in deep sediment communities; Rochelle PA et al.; A protocol was devised which permitted the extraction of DNA from deep marine sediments up to 503 m below the sea floor . These sediments have been laid down over the last 3 million years . 16S rRNA gene sequences were amplified from the DNA by the polymerase chain reaction . The details of the successful extraction and polymerase chain reaction methodology varied between samples from different depths . This emphasizes the attention to detail required to allow the diversity of bacteria in these deep sediments to be studied. Nucleic Acids Res, 1992 Dec 11, 20(23), 6235 - 8 Transcriptional regulation by folate: inducible gene expression in Dictyostelium transformants during growth and early development; Blusch J et al.; The Dictyostelium discoidin genes are induced in bacteria-grown cells shortly before the onset of development but are also highly expressed during growth in axenic medium . We here show that axenically growing cells strongly respond to the extracellular signal folate by suppressing discoidin synthesis while cell growth and development is not substantially affected . Repression occurs via two previously identified promoter elements, the dIE and the dAXE . Removal of the signal molecules or setting cells up for development results in rapid reactivation of the promoter . Based on this observation, we constructed the transformation vector pVEII and describe a convenient method which allows for controlled expression of a gene of interest in growing cells and also for external modulation in early development . Deletion constructs of the discoidin promoter can be used in addition to vary transcriptional activity over about one order of magnitude. J Immunol Methods, 1992 Dec 8, 156(2), 267 - 9 Use of passive immunization for the production of monoclonal antibodies against Actinobacillus pleuropneumoniae serotype 1,6 and 12; Stenbaek E; Hyperimmune sera from BALB/c mice immunized intraperitoneally (IP) with Actinobacillus pleuropneumoniae serotype 2 were used for passive intraperitoneal (i.p.) immunization of BALB/c mice . The immunized mice were subsequently immunized i.p . with a mixture of A . pleuropneumoniae serotypes 1, 6 and 12 . Numerous monoclonal antibodies specific for serotypes 1, 6 and 12 were obtained . Using this immunization scheme antibodies can be obtained against specific antigens from closely related bacteria. J Immunol Methods, 1992 Dec 8, 156(2), 163 - 70 Development of a monoclonal antibody to the conserved region of p34cdc2 protein kinase; Kamo K et al.; Mice and rabbits were injected with various forms of a 16 amino acid synthetic peptide representing PSTAIR, the evolutionarily conserved region of the protein kinase p34cdc2, for polyclonal antisera and hybridoma-monoclonal antibody production . Antisera from mice injected with an unconjugated monomeric form of the peptide showed no reaction to the peptide . Of four animals injected with the monomeric form of the peptide conjugated to keyhole limpet hemocyanin via m-maleimidobenzoyl-N-hydroxysulfosuccinimide (MBS), antisera from only one mouse had a very low titer to the peptide, and all four animals produced antibody to the MBS bridge . Both mice injected with an octameric multiple antigen peptide (MAP) of PSTAIR produced antisera reactive to the octameric MAP form of the peptide in ELISA and also to the cdc2 protein expressed in bacteria in an immunoblotting assay . Splenocytes from one mouse injected with the octameric MAP form of the peptide were successfully used for hybridoma-monoclonal antibody production . A monoclonal antibody was produced that reacted with octamer, monomer and cdc2-expressed protein and specifically with the carboxyl terminus of the 16 amino acid peptide. Ann N Y Acad Sci, 1992 Nov 21, 663, 139 - 53 Genotoxic-stress-response genes and growth-arrest genes . gadd, MyD, and other genes induced by treatments eliciting growth arrest; Fornace AJ Jr et al.; As discussed throughout this paper, many mammalian DDI genes are associated with growth responses, including both positive responses to growth stimulation and negative responses involving transient growth arrest and terminal differentiation . It is interesting that several immediate-early genes encoding transcription factors, the jun genes, are DDI, are induced by terminal differentiation, and also are associated with positive growth responses . In negative growth-response genes, their control is complex and almost certainly involves multiple regulatory mechanisms . The role of growth-arrest genes after exposure to DNA-damaging agents is currently not known, but as growth arrest can have a protective effect on cells exposed to DNA-damaging agents in both bacteria and eukaryotes, some protective role(s) for the gadd genes may exist . Whatever the roles are for the individual gadd genes, the response of the gadd genes to DNA-damaging agents and other growth-arrest signals has been highly conserved during mammalian evolution, and it is likely that this stress response, as reflected by induction of one or more gadd genes, is present in most or perhaps all mammalian cells . Our findings that the gadd group overlaps with another group of growth-arrest genes, the MyD, indicate that these two groups combined define a new class of genes whose protein products are likely to play a role in cell growth cessation. Med Hypotheses, 1992 Dec, 39(4), 323 - 33 Metabolism of the malignant cell, in vivo, is anaerobic and significantly plays a factor in the pathway to carcinogenesis; White MW; The malignant cell, in vivo, metabolizes and respires anaerobically . For a good many years scientists have been aware of anaerobia being present in malignant growths . The prevailing opinion is that the malignant cell is able to remain viable in an anaerobic medium . This assumption is incorrect and is actually misleading . My investigative studies indicates that the malignant cell metabolizes and respires intracellularly so that the consequence of such a physiology is anaerobic and the basis for such anaerobia is a metabolic phenomenon . There is a reason for this anaerobiosis . Studies demonstrated the fact that there are present within the malignant cell and in the immediate area bacterial spores arising from one of several varieties of plant bacteria . And with an adequate circulating flow of blood, by the animal (human) host, to that particular site, provides the metabolites, enzymes, and the exchange of gases to enhance this abnormal physiopathological anaerobiosis . It is this intracellular anaerobic metabolism within an animal cell that becomes the basis for its malignant transformation. Scott Med J . 1992 Dec;37(6):185. Abscess formation as a complication of chickenpox; Guthrie CM; Chickenpox is a common infectious disease of childhood . Skin lesions may occasionally become secondarily infected with bacteria . Large abscess formation is reported complicating a mild case of chickenpox in a previously healthy patient. Singapore Med J, 1992 Dec, 33(6), 570 - 4 Evaluation of therapies in the treatment of Helicobacter pylori associated non-ulcer dyspepsia; Nafeeza MI et al.; This randomised double blind placebo controlled study evaluated the effectiveness of colloidal bismuth subcitrate (CBS), ampicillin and their combination in the treatment of Helicobacter Pylori in non-ulcer dyspepsia (NUD) and assessed if elimination of this organism is associated with improvement of gastritis and the symptoms . Forty-eight NUD patients with H . pylori and histologic gastritis were randomly allocated to one of the three regimens for 28 days . Symptoms were assessed before and after treatment . Forty-three patients completed the trial . Repeat endoscopy within 48 hours of completing treatment showed suppression of H . pylori in 6 of 7 patients (85.7%) on combined therapy and one of 8 patients (12.5%) on CBS therapy (p = 0.0205) . There was no suppression of the bacteria in patients treated with ampicillin . Repeat endoscopy performed 2 weeks after completing treatment showed suppression of H . pylori in 3 of 7 patients (42.9%) on combined therapy and none in the other two groups . Patients on combined therapy who had suppression of H . pylori, 48 hours or 2 weeks after completing treatment were noted to have historical improvement of their gastritis (p = 0.0001 and p = 0.05 respectively) . This was also associated with improvement of symptoms in these patients. Rozhl Chir, 1992 Dec, 71(12), 677 - 82 {The modern approach to dressing wounds}; Sedlarik KM; The author evaluates from the pathophysiological aspect the impact of various dressings applied to wounds with regard to the permeability of secretions, gases, maintenance of a stable temperature and preservation of an optimum pH as well as with regard to preventing penetration of toxic substances, bacteria and foreign bodies . The author also pays attention to the important condition of non-adhesiveness of the dressing to the surface of the healing wound . From this aspect he evaluates cotton, paraffin, hydrocolloid, porous and silicon dressings . Based on the study of extensive literature and his own experiments, the author reaches the conclusion that it is essential to provide optimal mechanical and thermal protection of the wound and that for every type and stage of wound healing an appropriate dressing must be selected. J Card Surg, 1992 Dec, 7(4), 357 - 62 Simplified techniques of valve replacement; Cooley DA; After an extensive trial of many different suture techniques for implanting valvular prostheses, we have recently returned to using a simple, continuous suture of monofilament polypropylene . We believe this technique has many theoretical and practical advantages . Valvular implantation takes less time and is even more secure than when individual sutures are used . In addition, the need for porous felt pledgets, which may harbor bacteria, is eliminated . This simple technique facilitates implantation in the mitral and aortic positions. Anal Biochem, 1992 Dec, 207(2), 285 - 90 Alkylation of cysteine with acrylamide for protein sequence analysis; Brune DC; Alkylation of cysteine in proteins with acrylamide under mildly alkaline conditions yields a thioether derivative, Cys-S-beta-propionamide (Cys-S-Pam), which is stable during automated Edman degradation . Its phenylthiohydantoin derivative, PTH-Cys-S-Pam, is easily separated from other PTH-amino acids by HPLC and is thus useful for cysteine identification during protein sequencing . PTH-Cys-S-Pam was first noticed during sequencing polypeptides blotted onto polyvinylidene difluoride membranes from polyacrylamide gels, in which cysteine had reacted with residual unpolymerized acrylamide . Cysteine in proteins is easily alkylated by reaction of proteins in aqueous solution with acrylamide . Methods are also presented for alkylation of cysteine in proteins adsorbed on fiberglass disks in the reaction cartridge of a protein sequencer . Finally, PTH-Cys-S-Pam was synthesized chemically . The synthetic compound is unstable in neutral solution, but can be stabilized by acidification . It has the same HPLC retention time as the product formed from cysteine when sequencing proteins alkylated with acrylamide. Microbiol Rev, 1992 Dec, 56(4), 648 - 61 The antigen 85 complex: a major secretion product of Mycobacterium tuberculosis; Wiker HG et al.; The large number of different proteins synthesized by the mycobacterial cell are currently classified and studied in terms of groups of proteins with certain common properties such as physical and chemical characteristics, function, and localization in the mycobacterial cell . Proteins that are actively secreted during culture on synthetic media represent a particular group of great current interest . At least eight proteins secreted by Mycobacterium tuberculosis have been isolated and characterized to various extents . The genes coding for five proteins secreted from M . tuberculosis and/or Mycobacterium bovis BCG have been cloned and sequenced . All of them contain typical signal sequences . The proteins of the antigen 85 complex, which form the main subject of this review, are often the most common proteins in M . tuberculosis culture fluid . The constituents denoted 85A, 85B, and 85C are encoded by three genes located at different sites in the mycobacterial genome and show extensive cross-reactivity as well as homology at amino acid and gene levels . The proteins differ slightly in molecular mass in the 30- to 31-kDa region, and all of them are fibronectin-binding proteins, but the significance of the latter observation and the role of these proteins in mycobacterial physiology and interaction with the infected host remain to be elucidated . The antigen 85 complex proteins are strongly immunogenic in natural and experimental mycobacterial infections in terms of both induction of antibody synthesis and T-cell-mediated reactions . The well-recognized difference in the efficacy of live and dead mycobacterial vaccines should be considered in relation to the group of secreted antigens . After inoculation, live bacteria in vaccines such as BCG multiply in the host, probably releasing several constituents belonging to the class of secreted proteins and hence resulting in more efficient stimulation of the immune system . Secreted mycobacterial antigens are expected to be of particular significance in induction of various immune responses that are responsible for development of protective immunity in some individuals and for clinical symptoms and complications of the ensuing disease in others. J Lipid Res, 1992 Dec, 33(12), 1797 - 806 Fatty acid composition in deep hydrothermal vent symbiotic bivalves; Ben-Mlih F et al.; Fatty acids in deep hydrothermal vent bivalves have been analyzed . Their composition is completely different from that of a littoral mussel collected in the Mediterranean sea . The distribution of fatty acids in the littoral mussel is characterized by a predominance of polyunsaturated fatty acids (20:5n-3, 22:6n-3) reflecting the planktonic origin of the food . Vent bivalve fatty acid distribution is dominated by an abundance of the monounsaturated acids (double bond in the n-7 position) 16:1n-7, 18:1n-7, and 20:1n-7 which are clearly of bacterial origin and give an indication of the symbiotic bacterial activity in the bivalves . Differences between the fatty acid composition of the bivalves from two hydrothermal sites (13 degrees N and Galapagos) and differences between the mantle and the gill were observed and are discussed with respect to vent activities at the two sites and species metabolic capacities as a function of ecological conditions. FEMS Microbiol Rev, 1992 Dec, 9(2-4), 119 - 29 Biological complexes of poly-beta-hydroxybutyrate; Reusch RN; Short-chain complexed poly-beta-hydroxybutyrate, 130-170 monomer units, is a ubiquitous constituent of cells, wherein it is usually associated with other macromolecules by multiple coordinate bonds, or by hydrogen bonding and hydrophobic interactions . This conserved PHB has been isolated from the plasma membranes of bacteria, from a variety of plant tissues, and from the plasma membranes, mitochondria, and microsomes of animal cells . In bacterial membranes, PHB has been found complexed to the calcium salts of inorganic polyphosphates, and to single-stranded DNAs . The ability of PHB to solvate salts, consisting of cations having high solvation energies and large delocalized anions, is in accordance with its molecular characteristics, that of a flexible linear molecule possessing a large number of electron-donating ester oxygens suitably spaced to replace the hydration shell of cations . In turn, PHB may be rendered soluble in aqueous media by complexation to water-soluble proteins, such as serum lipoproteins and albumin . Such solvates are highly resistant to hydrolytic enzymes . These findings suggest that the physiological roles of this unique biopolymer may include the solvation of salts of polymeric anions to facilitate their movement through hydrophobic barriers, and the protection of cellular polymers from enzymatic degradation. Appl Environ Microbiol, 1992 Dec, 58(12), 3954 - 8 Resistance of proline-containing peptides to ruminal degradation in vitro; Yang CM et al.; Mixed ruminal bacteria utilized an enzymatic digest of casein at a rate faster than that for an enzymatic digest of gelatin, but neither amino acid source was completely utilized even when the incubation period was as long as 96 h . Since the reaction of ninhydrin with the residual nonammonia, nonprotein nitrogen was more than twofold stronger when the samples were hydrolyzed with 6 N HCl, it appeared that much of the residual nitrogen was from peptides . Approximately 66% of the nonammonia, nonprotein, ninhydrin-reactive material could not be recovered as amino acids, but there was a significant decrease in total amino acid nitrogen when the samples were pretreated with a C18 Sep-Pak column to remove peptides . The resistant peptides had an abundance of proline, and subsequent incubations showed that synthetic dipeptides which contained proline were hydrolyzed slowly . Lysine appears to be the amino acid which is most apt to limit ruminant production . Dipeptides containing proline and lysine were hydrolyzed at least fivefold slower than lysine-alanine . Methionine, another potentially limiting amino acid, was also degraded at a slower (2.5-fold) rate when it was present as part of a proline dipeptide. J Periodontol, 1992 Dec, 63(12), 974 - 8 Regenerative surgery of intrabony periodontal defects using ePTFE barrier membranes: scanning electron microscopic evaluation of retrieved membranes versus clinical healing; Selvig KA et al.; Sixteen intrabony defects in 12 patients were treated by gingival flap surgery including root surface debridement and placement of an expanded polytetrafluoroethylene (ePTFE) membrane . The membranes were removed after 4 to 6 weeks and examined by scanning electron microscopy (SEM) for bacterial contamination and adherent connective tissue elements . Twelve months postsurgery, the defect sites were reexamined for changes in probing attachment level and probing bone level . Comparison of ultrastructural findings and clinical observations revealed that extent of bacterial contamination of the membrane correlated inversely with clinical assessment of attachment gain . The results indicate that the extent of oral exposure and bacterial contamination of the ePTFE membrane at the time of removal may be an indicator of the long-term success or failure of the regenerative procedure. Neurosurgery, 1992 Dec, 31(6), 1132 - 5; 1134 e-PTFE ventricular shunt catheters; Gower DJ et al.; The perfect polymer surface for implantation should resist the adhesion of bacteria while producing no reaction in surrounding tissues . Silicone elastomer is a common material used for medical implantation devices . This material is superior to most available for implantation because of its flexibility and low, but detectable, tissue reaction . We have evaluated a flexible, expanded polytetrafluoroethylene (e-PTFE) ventricular catheter for cerebrospinal fluid shunting and have found that although e-PTFE is safe for cerebral implantation, the porosity of the material allows tissue ingrowth that obstructs the lumen . Our limited clinical evaluation of 5-microns internodal distance e-PTFE revealed that this open-cell structure, allowing tissue ingrowth, apparently is a poor candidate to replace silicone elastomer in cerebrospinal fluid shunting devices . However, further investigation may provide a polymer superior to silicone elastomer to create a cerebrospinal fluid shunt. Histopathology, 1992 Dec, 21(6), 521 - 8 Modes of Helicobacter colonization and gastric epithelial damage; Chan WY et al.; A total of 144 gastric biopsies colonized by Helicobacter-like organisms were studied under light and differential interference contrast microscopy for the modes of bacterial colonization . Biopsies were also graded for the degree of epithelial damage (epithelial-damage-grade: 0 to 6, in ascending order of severity) and density of Helicobacter-like organism (Helicobacter-grade: 0 to 6, in ascending order of bacterial density) . Three modes of colonization were identified: free-in-mucus, surface-adhesion and intercellular colonization . Because light microscopy cannot definitely prove the presence of intracellular colonization, bacteria located between cells and below the apical cell border were counted together as intercellular colonization . Bacteria free-in-mucus were seen in all biopsies . Surface adhesion was seen in 50-87.9% of biopsies, without obvious correlation with the epithelial-damage- and Helicobacter-grades . The incidences of intercellular and intracellular colonization were directly proportional to the epithelial-damage- and Helicobacter-grades . Free-in-mucus as the predominant mode of colonization was mainly seen in biopsies with lower (1-3) epithelial-damage- and Helicobacter-grades . Conversely, biopsies with intercellular colonization as the predominant mode of colonization were mainly cases with higher (4-6) epithelial-damage- and Helicobacter-grades . In cases showing predominantly bacteria between cells, 69.2% had a gastric ulcer whereas only 38.8% of cases showing predominantly bacteria free-in-mucus showed ulceration (P < 0.01) . These results indicate that Helicobacter-like organisms can invade and penetrate between epithelial cells . When free-in-mucus, Helicobacter-like organisms are less likely to induce epithelial damage . However, the more invasive modes of colonization (intercellular) were associated with severe epithelial damage and high Helicobacter density.(ABSTRACT TRUNCATED AT 250 WORDS) Am J Clin Pathol, 1992 Dec, 98(6), 569 - 74 Helicobacter pylori-related gastritis and gastric ulcer . A continuum of progressive epithelial degeneration; Leung KM et al.; One hundred forty-five consecutive gastric biopsy specimens showing colonization by Helicobacter pylori (HP) were studied . Biopsy specimens were obtained from patients with the following conditions: gastric ulcer (GU; 76), active chronic gastritis (ACG; 52), GU with duodenal ulcer (DU; 10), and ACG with DU (7) . The mean age of the patients in the ACG group was 8.6 years less than the patients in the GU group . Helicobacter pylori colonization and HP-induced epithelial degeneration (ED) were quantified by a grading system (grades 0 to 6) comprising both focal and global scores for bacterial density (HP grade) and severity of ED (ED grade) . The ED grade was directly proportional to the HP grade in all biopsy specimens . Gastric ulcer biopsy specimens were associated with higher HP grades: HP grade more than 5 in 25 cases (32.9%) and ED grade more than 5 in 18 cases (23.6%) of GU compared with similar respective scores in 9 cases (17.9%) and 2 cases (3.8%) of ACG . The difference was due primarily to a higher global score of bacterial density and higher focal score of ED in the GU biopsy specimens . These results support the hypothesis that HP-positive ACG and HP-positive GU are lesions within a single disease spectrum . Heavy HP colonization and severe HP-induced epithelial damage are predisposing factors in ulcerogenesis . Because HP-positive ACG is probably a preulcerative state, eradication of the bacteria in HP-positive ACG might prevent subsequent GU. Obstet Gynecol, 1992 Dec, 80(6), 1017 - 9 Effect of time and storage temperature on amniotic fluid glucose concentration; Meyer WJ et al.; OBJECTIVE: To evaluate the effects of time and storage temperature on amniotic fluid (AF) glucose concentration . METHODS: Amniocentesis was performed on 47 patients with preterm labor or preterm rupture of membranes at or before 34 weeks' estimated gestation and the AF was cultured for aerobic, anaerobic, Mycoplasma, and Ureaplasma organisms . Twenty-one samples were stored at 37C, 13 were kept at room temperature (22C), and 13 were frozen (-20C) . Glucose concentration was measured at 0 and 12 hours on all AF samples and at 2, 4, and 6 hours on the unfrozen samples . RESULTS: No significant change in AF glucose concentration occurred over time at any storage temperature in the 35 samples with negative cultures . In 12 AF samples with positive cultures, a significant decrease from baseline was noted at 2 and 4 hours in those kept at 37C and 22C (P = .016) . Glucose concentration was unchanged in frozen samples regardless of the culture results . The initial AF glucose concentration was significantly lower in samples with positive than in those with negative cultures (P < .0001) . CONCLUSION: Amniotic fluid glucose concentration does not change over 12 hours in the absence of intra-amniotic infection. J Bacteriol, 1992 Dec, 174(23), 7595 - 605 Identification, isolation, and structural studies of the outer membrane lipopolysaccharide of Caulobacter crescentus; Ravenscroft N et al.; The lipopolysaccharide (LPS) of the outer membrane of Caulobacter crescentus was purified and analyzed . Two distinct strains of the species, NA 1000 and CB2A, were examined; despite differences in other membrane-related polysaccharides, the two gave similar LPS composition profiles . The LPS was the equivalent of the rough LPS described for other bacteria in that it lacked the ladder of polysaccharide-containing species that results from addition of variable amounts of a repeated sequence of sugars, as detected by gel electrophoresis in smooth LPS strains . The purified LPS contained two definable regions: (i) an oligosaccharide region, consisting of an inner core of three residues of 2-keto-3-deoxyoctonate, two residues of alpha-L-glycero-D-mannoheptose, and one alpha-D-glycero-D-mannoheptose unit and an outer core region containing one residue each of alpha-D-mannose, alpha-D-galactose, and alpha-D-glucose, with the glucose likely phosphorylated and (ii) a region equivalent to the lipid A region of the archetype, consisting primarily of an esterified fatty acid, 3-OH-dodecanoate . The lipid A-like region was resistant to conclusive analysis; in particular, although a variety of analytical methods were used, no amino sugars were detected, as is found in the lipid A of the LPS of most bacteria. J Virol, 1992 Dec, 66(12), 7113 - 20 Baculovirus phosphoprotein pp31 is associated with virogenic stroma; Guarino LA et al.; The PstI K fragment of Autographa californica nuclear polyhedrosis virus (AcMNPV) encodes a protein with a molecular weight of 31,000 . To define the role of this protein (pp31) in virus infection further, it was overexpressed in bacteria and used to produce polyclonal antiserum . Radioimmunoprecipitation analysis indicated that pp31 was synthesized during both the early and late phases of virus infection, consistent with previous analyses indicating that the gene was regulated by tandem early and late promoters . Metabolic labeling of cells with carrier-free phosphate indicated that pp31 was phosphorylated . Biochemical fractionation experiments showed that pp31 was localized in the nucleus and that it was more stably associated with the nucleus at later times of infection . Immunoblot analysis of subnuclear fractions indicated that pp31 was associated predominantly with the chromatin and nuclear matrix fractions . Immunofluorescence experiments confirmed that the pp31 protein was localized in the nucleus . Nuclear staining was relatively uniform early but was more centrally nuclear later in infection . Immunoelectron microscopy indicated that the pp31 protein was a component of virogenic stroma . Southwestern (DNA-protein) blot analysis demonstrated that pp31 is a DNA-binding protein . These findings suggest a possible role for pp31 in the virus life cycle. J Infect Dis, 1992 Dec, 166(6), 1449 - 53 A subpopulation of Treponema pallidum is resistant to phagocytosis: possible mechanism of persistence; Lukehart SA et al.; While untreated syphilis infection is characterized by spontaneous resolution of early lesions, a few organisms evade the host immune response and persist for many years . Macrophages are generally recognized as the effector cell responsible for bacterial clearance, and phagocytosis is enhanced by immune serum . This study examined the susceptibility of Treponema pallidum isolated at various stages of lesion resolution to opsonization and phagocytosis by macrophages in vitro . Findings suggest that the population of organisms remaining after the majority of bacteria have been cleared in vivo is resistant to phagocytosis . This may provide a mechanism for the persistence of T . pallidum in the face of an otherwise active immune response. APMIS, 1992 Dec, 100(12), 1066 - 72 Fimbriae of Branhamella catarrhalis as possible mediators of adherence to pharyngeal epithelial cells; Ahmed K; This study attempted to elucidate the role of fimbriae in the adherence of B . catarrhalis to human oropharyngeal epithelial cells . Antifimbrial immune serum was prepared by immunization of rabbit with whole fimbriated bacteria, and adsorption of the serum with a nonfimbriated B . catarrhalis strain . After pretreatment with the antifimbrial antiserum, the adherence of fimbriated B . catarrhalis to human epithelial cells was significantly decreased (p < 0.05) . The adherence was also significantly (p < 0.001) decreased by trypsin treatment . Electron microscopy revealed destruction of fimbriae after trypsin treatment . These observations suggest that fimbriae are involved in the adherence of B . catarrhalis to epithelial cells. Infect Agents Dis, 1992 Dec, 1(6), 279 - 93 Challenge of Chlamydia research; Stephens RS; Chlamydia is an obligate intracellular bacterial pathogen that severely challenges the patience and creativity of all its investigators--even to the point that some investigators have forsaken this field for more productive and fertile areas of research . The two principal difficulties that touch every aspect of chlamydial research are (a) that chlamydiae only grow within eukaryotic host cells and (b) there are limited genetic approaches available . Despite these technical difficulties, the fundamental underlying problem has been the expectation that chlamydiae are similar to other bacteria (or, historically, viruses) and amenable to study from this perspective . However, this has often turned out not to be the case . Chlamydiae have shown themselves to be unique at many levels and thus represent a formidable, yet enticing, research challenge. Zhonghua Jie He He Hu Xi Za Zhi, 1992 Dec, 15(6), 339 - 40, 379 {Laboratory study and clinical evaluation of ABC-ELISA for detection of mycoplasma pneumoniae antigens}; Yu KL; Using anti-M . pneumoniae IgG obtained by immunizing rabbits with whole cell antigen of M . pneumoniae strain FH, we established an ABC-ELISA for detection of M . pneumoniae in simulated positive specimens, ABC-ELISA had a detection limit of 10(4) cfu/ml of specimen . There were no cross-reaction with 4 other species of human mycoplasmas and 12 common species of bacteria in respiratory tract . In 47 patients suspected of being M . pneumoniae infection, we compared the results of ABC-ELISA with the combined results from serodiagnostic methods and M . pneumoniae culture . The coincident rate between them was 80.9%. Oral Microbiol Immunol, 1992 Dec, 7(6), 364 - 7 Interaction of Porphyromonas gingivalis with gingival epithelial cells maintained in culture; Lamont RJ et al.; Interactions between Porphyromonas gingivalis and gingival epithelial cells were investigated . Gingival epithelial cells were cultured from surgically removed gingival tissue . Electron microscopy demonstrated adherence of P . gingivalis to the cell membranes and microvilli followed by internalization of the bacteria into the epithelial cell cytoplasm . Saliva from healthy and periodontally diseased patients inhibited P . gingivalis association with the epithelial cells . Attachment to and penetration of gingival epithelial cells by P . gingivalis may be important virulence factors in periodontal disease . Salivary molecules may play a role in modulating these interactions. Exp Hematol, 1992 Dec, 20(11), 1278 - 84 Photodamaging effects of merocyanine 540 on neutrophils and HL-60 cells; Smith OM et al.; Merocyanine 540 (MC540) is a photosensitizing dye that has been used in several preclinical models and in a phase I clinical trial for the extracorporeal purging of tumor cells from autologous bone marrow grafts . The mechanism of the cytotoxic activity of MC540 is not yet fully understood, and the subcellular targets of MC540-mediated photodynamic damage remain to be identified . The human neutrophil provides an attractive model with which to study the effects of photoactivated MC540 on several well-defined cellular functions . As we report in this paper, simultaneous exposure of neutrophils to MC540 and light inhibited phagocytosis, random migration, chemotaxis, hydrogen peroxide production, and oxygen consumption . By contrast, the ability of neutrophils to kill engulfed bacteria and to produce superoxide radical was not compromised . Intracellular ATP levels and the activities of the cytosolic enzymes superoxide dismutase, catalase, and myeloperoxidase were only slightly reduced . Even in HL-60 leukemia cells, which bind more dye and are more readily killed by MC540-mediated photodynamic therapy than neutrophils, superoxide dismutase, catalase, and myeloperoxidase activities remained at normal or near-normal levels . These results are compatible with the view that plasma membrane components are primary targets of MC540-mediated photodynamic damage. J Clin Invest, 1992 Dec, 90(6), 2197 - 208 Sequence analyses of three immunoglobulin G anti-virus antibodies reveal their utilization of autoantibody-related immunoglobulin Vh genes, but not V lambda genes; Huang DF et al.; Accumulated sequence analyses of the antibody repertoire have revealed that most autoantibodies and developmentally regulated antibodies share a small set of germline Ig-variable region (V) genes . The findings have prompted speculation that certain autoantibodies are of developmental importance and may be instrumental in maintaining homeostasis of the adult antibody repertoire . In order to evaluate this hypothesis critically, it is first necessary to determine the V gene usage in human antibodies against foreign substances . Unfortunately, only a few such antibodies have had their heavy and light chains characterized . To rectify the situation, we adapted the anchored polymerase chain reaction to clone and analyze rapidly the expressed V genes for three anti-virus IgG antibodies . The results show that all three heavy chain V (Vh) genes are highly homologous to the known autoantibody-related Vh genes . In contrast, two light chain V (VL) genes of the V lambda 1 subgroup are similar to a non-autoantibody-related germline V lambda 1 gene . Taken together with the reported Vh and VL sequences of several antibodies against viruses and bacteria, the data show that many antipathogen antibodies may use the same small set of Vh genes that encode autoantibodies, but diverse VL genes that are distinct from autoantibody-related VL genes . Thus, only a small portion of the potentially functional germline Vh genes are used recurrently to generate most antibodies in a normal antibody repertoire, regardless of their reactivities with either self or non-self. Yan Ke Xue Bao, 1992 Dec, 8(4), 165 - 8, 164 {Vitrectomy combined with intravitreal injection of drugs for the management of advanced suppurative endophthalmitis}; Wen X et al.; Sixty cases (60 eyes) of advanced suppurative endophthalmitis treated by a combination of vitrectomy and intravitreal injection of drugs (antibiotics or/and corticosteroids) were investigated perspectively . Of the 60 cases, 43 were infected after trauma, 12 were infected after surgical procedures, while the others were of unknown causes . During the follow-up period of 1/2-5 years, 57 cases (95%) retained functioning eyes, the postoperative visual acuity of 34 eyes (57%) being 0.05 or better and the postoperative visual acuity of 17 eyes (28%) 0.4 to 1.0 . The advantages of the therapeutic regimen are analyzed . We believe that it is the most effective therapy for suppurative endophthalmitis . The timing of the operation and the factors influencing the results are discussed . This report also presents our clinical experiences in the management of advanced suppurative endophthalmitis. Glycoconj J, 1992 Dec, 9(6), 325 - 9 Glycosphingolipid expression in spontaneously aborted fetuses and placenta from blood group p women . Evidence for placenta being the primary target for anti-Tja-antibodies; Lindstrom K et al.; A 12-week-old fetus and one 17-week-old fetus + placenta were obtained after spontaneous abortions from two women of blood group p . The 17-week-old fetus was dissected into intestine, liver, brain and residual tissue . Nonacid glycosphingolipid fractions were prepared from the tissues . Glycolipid characterization was carried out using thin layer chromatography immunostained with monoclonal antibodies and bacteria and by 1H NMR spectroscopy and mass spectrometry . In the placental fraction substantial amounts of globotetraosylceramide (P-antigen) and globotriaosylceramide (Pk-antigen) were identified . In contrast, the fetuses contained only trace amounts of these structures, as revealed by immunostaining . These results indicate that the primary target for the antibodies of the anti-Tja serum is the placenta tissue, resulting in termination of the pregnancy. Cell Struct Funct, 1992 Dec, 17(6), 417 - 25 Correlation of axenic linkage groups with the position of the microtubule-organizing center in aggregating Dictyostelium; Sameshima M et al.; Positioning of the microtubule-organizing center (MTOC) in Dictyostelium discoideum was found to be genetically regulated . We examined the wild-type strain NC-4 cells independently maintained in different laboratories, freshly recovered cells from spores stocked for over 20 years, the temperature-sensitive growth mutant HU49 isolated from NC-4, as well as strain V-12 which is the opposite mating-type to NC-4 . During aggregation on nonnutrient agar plates, all these strains showed similar cell polarity, as defined by the alignment of the nucleus ahead of the MTOC . By contrast, in Ax2 and Ax3, axenic strains carrying axenic mutations on linkage groups II and III, the MTOC was usually positioned ahead of the nucleus . Cells containing axenic linkage group II but not III positioned the MTOC ahead of the nucleus . Conversely cell polarity of strains including axenic linkage group III but not II was similar to that of wild-type cells . Thus axenic linkage group II, probably axeC or other linked gene(s) not yet identified, is responsible for the location of the MTOC anterior to the nucleus during aggregation . The anterior positioning of the MTOCs was prevented by growth on bacteria in cells carrying both axenic linkage groups, but not in those carrying only axenic linkage group II. Kekkaku, 1992 Dec, 67(12), 787 - 94 {Polymerase chain reaction for identification of Mycobacterium in sputa}; Arai T et al.; Polymerase chain reaction was applied to the diagnosis of mycobacterial infections . This reaction could detect bacteria in clinical specimens within a few hours . Sensitivities of the five primer systems reported in 1990 were compared . It was proved that the primer system reported by Eisenach et al . was as sensitive as culture on Ogawa's medium for the detection of Mycobacterium tuberculosis in sputa . Other systems were found to be less sensitive than this system, and the nested PCR should be applied to make these systems highly sensitive . For the practical application of PCR method, we should improve the detection system to fit for the practice in clinical laboratory . High cost of PCR system could be another barrier for the practical application. Indian J Biochem Biophys, 1992 Dec, 29(6), 508 - 11 Purification of C-reactive protein from Channa punctatus (Bloch); Mitra S et al.; C-reactive protein (CRP) is found to be a normal component of serum of freshwater air-breathing murrel Channa punctatus . Based on the property of binding with C-polysaccharide (CPS) of pneumococcus bacteria in presence of Ca2+, CRP was purified by phosphorylcholine-Sepharose affinity column chromatography . Molecular weight of the intact protein molecule was estimated to be approximately 141,000 by gel filtration . In non-reduced and reduced conditions the molecule showed molecular weight approximately 28,000 and 14,000 respectively in SDS-PAGE . Monospecific antisera was raised against the affinity purified CRP and used as a tool to detect CRP in the hepatic cytosol and egg extract . The level of CRP in the normal serum was estimated to be 220 micrograms/ml. Biokhimiia, 1992 Dec, 57(12), 1862 - 73 {Uneven distribution of amino acid substitutions throughout the amino acid sequence of homologous proteins}; Kostetskii PV; A set of aligned homologous protein sequences is divided into two groups consisting of m and n most related sequences . The value of position variability for homologous protein sequences is defined as a number of failures to coincide in the intergroup comparison of all possible m*n pairs of amino acid residues in that position divided by m*n . The position variability value plotted versus the sequence position number with a window of 10 positions gives the intergroup local variability profile . Area S of the figure included between the local variability profile and the straight line corresponding to the mean local variability value is compared with the average area Sr for 1000 random homologous protein families . If S is greater than Sr by more than 2 standard deviation units sigma r, the local variability profile is assumed to contain peaks and hollows corresponding to significant variable and conservative regions of the sequences . The profile extrema containing the area surplus delta S = S-(Sr+ 2 sigma r) are cut off by two straight lines to locate significant regions . The difference (S-Sr) given in standard deviation units sigma r is believed to be the amino acid substitution overall irregularity along the homologous protein sequences OI = (S-Sr)/sigma r . The significant conservative and variable regions of six homologous sequence families (phospholipase A2, cytochromes b, alpha-subunits of Na,K-ATPase, L- and M-subunits of photosynthetic bacteria photoreaction centre and human rhodopsins) were identified . It was shown that for artificial homologous protein sequences derived by k-fold lengthening of natural protein sequences, the OI value rises as square root of k . To compare the degree of substitution irregularity in homologous protein sequence families of different lengths L the value of standard substitution overall irregularity for L = 250 is proposed. Neuroreport, 1992 Dec, 3(12), 1073 - 6 Polyclonal antibodies against the rat NK1 receptor: characterization and localization in the spinal cord; Moussaoui SM et al.; We have developed antibodies against the NK1 receptor and have investigated its cellular distribution . Rabbit polyclonal antibodies were generated against peptide (19-32) of the rat brain NK1 receptor . They were very specific to the NK1 site as shown by ELISA against various epitopes of NK1, NK2 and NK3 receptors and by immunoblotting of proteins from bacteria transfected with rat brain NK1 receptor cDNA and from rat cortex . Determining how immunostained NK1 receptors are distributed in the rat spinal cord made it possible to identify the cellular structures on which NK1 receptors are located and where they form synapses with SP terminals . In the superficial layers of the dorsal horn, the NK1 receptors appeared mainly of dendritic nature and were, like SP, abundant . In the deep layers of the dorsal horn and in the ventral horn, they were associated mostly with cell bodies. J Neurosci Res, 1992 Dec, 33(4), 639 - 48 L2/HNK-1 carbohydrate and protein-protein interactions mediate the homophilic binding of the neural adhesion molecule P0; Griffith LS et al.; The neural adhesion molecule P0, the most abundant glycoprotein in peripheral myelin of mammals, is a member of the immunoglobulin superfamily and expresses the L2/HNK-1 and L3 oligosaccharides at a single N-glycosylation site . It acts in both homophilic and heterophilic binding mechanisms . To investigate the molecular requirements for homophilic interaction, we have used P0 from human sciatic nerve and the extracellular domain of P0 expressed in bacteria to determine binding of P0 to P0 in solid phase and bead aggregation assays . The binding of P0 to P0 could be partially inhibited in both assays by antibodies to the L2/HNK-1 epitope and by the L2/HNK-1 carbohydrate, but not by L3 antibodies or other carbohydrates . Inhibition of binding was also seen with polyclonal antibodies reacting with the protein backbone of P0 . These observations indicate that both carbohydrate and protein structures are involved in the binding of P0 to P0 and that P0 acts as a presenter of and a receptor for a functionally important carbohydrate. Mutat Res, 1992 Dec, 298(2), 71 - 9 Mutagenicity testing of imidazole and related compounds; Forster R et al.; Ames tests have been performed with imidazole and its principal metabolites, hydantoin and hydantoic acid . N-Acetyl-imidazole, a potential metabolite resulting from the action of intestinal bacteria, and histamine, a structurally related compound which is widely distributed in mammalian tissues, have also been tested . Imidazole and histamine were also tested in the UDS assay in primary rat hepatocytes, while imidazole alone was tested in the M2-C3H mouse fibroblast malignant transformation assay . Imidazole gave consistently negative results in the Ames test, the UDS assay and the transformation assay . The three metabolites of imidazole, namely hydantoin, hydantoic acid and N-acetyl-imidazole, all gave negative results in the Ames test . Histamine gave no evidence of mutagenic activity in the Ames test or of genotoxicity in the UDS assay . These results indicate that imidazole and its metabolites are unlikely to present a mutagenic or carcinogenic hazard. J Mol Evol, 1992 Dec, 35(6), 551 - 6 A redefinition of the Asp-Asp domain of reverse transcriptases; Velasco AM et al.; The rules defining the Asp-Asp domain of RNA-dependent polymerases deduced by Argos (1988) were tested in a set of 53 putative reverse transcriptases (RTs) sequences . Since it was found that some of these rules are not followed by RTs coded by bacteria, group II introns, and non-LTR retrotransposons, we present here a more strict definition of the Asp-Asp domain. Fortschr Med, 1992 Nov 30, 110(30), 565 - 70 {Prevention of recurrent infection of the upper and lower airways . Multicenter, open study over three months}; Debelic M et al.; In a multicenter open study involving 620 patients suffering from recurrent infections of the upper and lower respiratory tract aged between 12 and 89 years, the tolerability of an oral bacterial lysate was investigated . On the basis of the nature and frequency (1.9%) of adverse effects, and an analysis of the laboratory investigations over an observation period of three months, overall tolerability was adjudged to be excellent . The frequency and severity of infections was reduced . In 75% of the cases, both physician and patient considered the augmentation of the body's defense mechanisms by immunoactivation to have been of benefit . The favourable risk-to-benefit ratio results in a high level of acceptance of immunotherapy, and 96% of the patients completed the study as planned. J Mol Biol, 1992 Nov 20, 228(2), 698 - 700 A myoglobin evolved from indoleamine 2,3-dioxygenase; Suzuki T et al.; Hemoglobins and myoglobins are some of the best studied proteins . They are distributed in animals, plants and bacteria, and the characteristic two intron-three exon structure is widely conserved in animal globin genes (Jhiang et al., 1988) . To date, all of the hemoglobins and myoglobins are believed to have a common origin, and so they are considered to be homologous . We have isolated a completely new type of myoglobin from the red muscle of the abalone Sulculus diversicolor aquatilis . The myoglobin consists of an unusual 41 kDa polypeptide chain, contains one heme per chain and forms a homodimer under physiological conditions . The cDNA-derived amino acid sequence of Sulculus myoglobin showed no significant homology with any other globins, but, surprisingly, showed high homology (35% identity) with human indoleamine 2,3-dioxygenase, a tryptophan degrading enzyme containing heme . This clearly indicates that Sulculus myoglobin evolved from a gene for indoleamine dioxygenase, but not from a globin gene . Sulculus myoglobin lacks the enzyme activity of indoleamine dioxygenase . However, in the presence of tryptophan, the autoxidation rate of oxymyoglobin was greatly accelerated, suggesting that a tryptophan binding site remains near or in the heme cavity as a relic of the molecular evolution. FEBS Lett, 1992 Nov 9, 312(2-3), 115 - 22 Identification, characterization, and molecular cloning of a novel transporter-like protein localized to the central nervous system; Gingrich JA et al.; During the course of large scale purification of the D1 dopamine receptor from rat brain, a protein of approximately 87,000 daltons (p87) was observed to copurify with the D1 receptor through four chromatographic steps . To characterize the nature of this protein, bovine and rat cDNA clones were isolated and sequenced . The bovine and rat clones were highly conserved (98.5% identity) . Each clone possessed an open reading frame of 2226 base pairs encoding a protein of 742 amino acids (calculated MW of 82,500), containing three stretches of peptide sequence obtained from p87 sequence analysis . Comparison of the deduced peptide sequence of this protein with those found in available databanks revealed that it was a novel protein related to the family of nutrient transport proteins from eukaryotes and bacteria, including, the mammalian facilitated glucose transporters, the yeast transporters for maltose, lactose, and glucose, and the proton-driven bacterial transporters for arabinose, xylose, and citrate . In addition p87 also shares with these transporters a similar hydropathicity profile that suggests the presence of 12 transmembrane segments . The mRNA for p87 appears to be localized primarily, if not exclusively, to the central nervous system . Northern blot analysis reveals a message of approximately 4.8 kb in cortex, hippocampus, brain stem, and cerebellum, but no detectable signal in peripheral tissues such as spleen, liver, kidney, lung, heart, or skeletal muscle . Evidence form Western blot analysis and immunohistochemistry suggests that this protein may be expressed in intracellular organelles or the membrane of synaptosomes rather than plasma membrane . Based on its structure and properties, p87 appears to define a new class of transporter-like proteins. Anal Biochem, 1992 Nov 15, 207(1), 142 - 9 Application of an immune-tolerizing procedure to generate monoclonal antibodies specific to an alternate protein isoform of bovine growth hormone; Salata RA et al.; An immune-tolerizing protocol was employed to generate monoclonal antibodies to a variant protein isoform of bovine growth hormone arising from alternative pre-mRNA processing . Variant bovine growth hormone used for immunization was obtained by expression in bacteria and electroelution of the protein from preparative sodium dodecyl sulfate-polyacrylamide gels . Balb/c mice were first immunized with wild-type bovine growth hormone in the presence of the cytotoxic drug cyclophosphamide, thereby tolerizing the mouse to common epitopes shared among the two proteins . Subsequently, the mice were immunized with variant bovine growth hormone to produce antibodies specific to variant epitopes . Comparisons of fusions resulting from standard and tolerizing immunization protocols resulted in a significantly enhanced production of variant bovine growth hormone-specific antibodies as a result of the immunotolerizing protocol . The specificity of the antibodies to the variant growth hormone was substantiated by differential enzyme-linked immunosorbent assay and Western blot . Nearly all hybridomas positive for variant growth hormone were negative for wild-type growth hormone . Finally, the antibodies were used to demonstrate intracytoplasmic staining of COS I cells transiently transfected with a variant growth hormone-producing plasmid . Given the power of the polymerase chain reaction to conveniently clone alternatively processed mRNA species, followed by expression in bacteria to provide antigen, the immunotolerizing protocol provides a convenient general method for producing antibodies specific to desired protein isoforms. Proc Natl Acad Sci U S A, 1992 Nov 15, 89(22), 10887 - 91 Cloning of a human galactokinase gene (GK2) on chromosome 15 by complementation in yeast; Lee RT et al.; A human cDNA encoding a galactokinase (EC 2.7.1.6) was isolated by complementation of a galactokinase-deficient (gal1-) strain of Saccharomyces cerevisiae . This cDNA encodes a predicted protein of 458 amino acids with 29% identity to galactokinase of Saccharomyces carlsbergensis . Previous studies have mapped a human galactokinase gene (GK1) to chromosome 17q23-25, closely linked to thymidine kinase . The galactokinase gene that we have isolated (GK2) is located on chromosome 15 . The relationship between the disease locus for galactokinase deficiency galactosemia, which is responsible for cataracts in newborns and possibly presenile cataracts in adults, and the two galactokinase loci is unknown. J Biol Chem, 1992 Nov 15, 267(32), 23057 - 62 Dominant negative mutants of ornithine decarboxylase; Tsirka S et al.; Conserved lysines of mouse ornithine decarboxylase were individually mutated to arginines . The mutations at amino acid residues 69, 115, and 169 greatly reduced or abolished enzymatic activity . Lysine 69 is the site of Schiff base formation with the cofactor pyridoxal phosphate; the functional role of the other two lysines essential for activity is not known . Coexpression of wild type ornithine decarboxylase along with the lysine 115 to arginine mutant reduced the activity of the former without diminishing the amount of wild type protein . This form of negative complementation was seen when wild type and mutant protein were coexpressed either by in vitro translation or in bacteria . The data are consistent with the conclusion that a wild type and mutant subunit form a heterodimer that is enzymatically inactive. J Clin Microbiol, 1992 Nov, 30(11), 2788 - 92 Use of a recombinant 170-kilodalton surface antigen of Entamoeba histolytica for serodiagnosis of amebiasis and identification of immunodominant domains of the native molecule; Zhang Y et al.; We expressed the gene that encodes one of the major surface antigens of Entamoeba histolytica, the 170-kDa protein (1,270 amino acids), as a glutathione S-transferase fusion protein containing amino acids 1 to 1202 (lacking the putative transmembrane and cytoplasmic regions) and as separate fusion proteins containing each of three major domains of the 170-kDa molecule . Lysates from bacteria induced to express one of these proteins were used as the target antigens in a Western blot (immunoblot) analysis to determine whether a recombinant 170-kDa antigen could serve as the basis for a serologic test used to detect invasive amebiasis and whether there are differences in humoral immunogenicity among the three major domains of the 170-kDa antigen . Among patients with invasive amebiasis from three major areas where the disease is endemic and two sites in the United States, 54 (90%) of 60 had antibodies to the recombinant 170-kDa protein . Among 37 patients from regions where the disease is endemic and 20 patients from the United States without amebic disease, 1 (2%) of 57 had antibodies to the recombinant 170-kDa protein . We found significant differences in seroreactivity to each of three major domains of the molecule among patients seropositive for the complete construct, ranging from 100% seroreactivity with the fusion protein containing the domain designated cysteine rich and 89% seropositivity with the fusion protein incorporating a portion of the region designated cysteine poor to only 9% seropositivity for the fusion protein containing the pseudorepeat domain . Our study indicates that a serologic test based on the recombinant 170-kDA antigen could serve as a highly sensitive and specific test for acute invasive amebiasis. Nucleic Acids Res, 1992 Nov 11, 20(21), 5587 - 92 Isolation of cDNA clones encoding the beta isozyme of human DNA topoisomerase II and localisation of the gene to chromosome 3p24; Jenkins JR et al.; Topoisomerases catalyse the interconversion of topological isomers of DNA and have key roles in nucleic acid metabolism . Human cells express two distinct type II topoisomerase isozymes, designated topoisomerase II alpha (170 kDa form) and topoisomerase II beta (180 kDa form) . We have isolated cDNA clones encoding the beta isozyme from a human B-cell library . The proposed coding region for the topoisomerase II beta protein is 4,863 nucleotides long and would encode a polypeptide with a calculated M(r) of 182,705 . The predicted topoisomerase II beta protein sequence shows striking similarity (72% identical residues) to that of the human alpha isozyme, and homology to topoisomerase II proteins from Drosophila, yeast and bacteria . Regions of greatest amino acid sequence divergence lie at the extreme N-terminus and over a C-terminal domain comprising approximately 25% of the total protein . We have quantified the level of topoisomerase II beta mRNA in a panel of human tumour cell lines of different origin using an RNase protection assay, and compared the level to that of topoisomerase II alpha mRNA . Topoisomerase II beta mRNA was expressed in haemopoietic, epithelial and fibroblast cell lines, although to different extents, with U937 cells (promonocytic leukaemia) showing a particularly high level . There was no obvious relationship in terms of level of expression between the topoisomerase II alpha and beta genes . We have localised the gene encoding topoisomerase II beta protein to chromosome 3p24 in the human genome. J Chromatogr, 1992 Nov 6, 582(1-2), 242 - 5 Ion-pair high-performance liquid chromatography of diaminopimelic acid in hydrolysates of physiological samples; Dugan ME et al.; A method is reported for the determination of diaminopimelic acid (DAPA) in physiological samples . DAPA is derivatized with an o-phthaldialdehyde reagent solution, subjected to reversed-phase high-performance liquid chromatography and detected spectrofluorometrically . The method is a significant advance over previous methods because it uses the ion-pairing agent hexadecyltrimethylammonium bromide (HTMA) to facilitate DAPA measurement . Ion-pairing with HTMA avoids interference with co-eluting derivatives to provide simultaneous, sensitive, reproducible measurement of both DAPA peaks (DD,LL-DAPA and DL-DAPA). Science, 1992 Nov 6, 258(5084), 931 - 6 Transport of proteins across the endoplasmic reticulum membrane; Rapoport TA; The biosynthesis of many eukaryotic proteins requires their transport across the endoplasmic reticulum (ER) membrane . The process can be divided into two phases: (i) a targeting cycle, during which, by virtue of their signal sequences, nascent polypeptides are directed to translocation sites in the ER and (ii) the actual transfer of proteins across the membrane . The first phase has been well characterized, whereas the latter until recently was completely unresolved . Key components of the translocation apparatus have now been identified and it seems likely that they form a protein-conducting channel in the ER membrane . The transport process is similar to the process of protein export in bacteria. J Mol Biol, 1992 Nov 5, 228(1), 302 - 5 Crystallization and preliminary crystallographic investigation of methanol dehydrogenase from Methylobacterium extorquens AM1; Ghosh M et al.; Single crystals of methanol dehydrogenase (MDH) from Methylobacterium extorquens AM1 have been grown by the vapour diffusion method . These crystals diffract to beyond 2 A resolution and are suitable for X-ray crystallography . They belong to the orthorhombic space group P2(1)2(1)2(1) and have the following unit cell parameters: a = 66.79 A, b = 108.9 A, c = 188.9 A . One asymmetric unit contains an alpha 2 beta 2 tetramer of MDH and the location of the non-crystallographic 2-fold symmetry axis of this tetramer is defined by the paired positions of the binding sites of heavy atoms in four MDH-derivatives. Biochemistry, 1992 Nov 3, 31(43), 10529 - 35 Enzymatic properties of the RecA803 protein, a partial suppressor of recF mutations; Madiraju MV et al.; The RecA803 protein suppresses the recombinational repair defect of recF mutations and displays enhanced joint molecule formation in vitro (Madiraju et al