Microb Pathog, 1987 Jan, 2(1), 3 - 14 Experimental murine tularemia caused by Francisella tularensis, live vaccine strain: a model of acquired cellular resistance; Anthony LS et al.; We have established a model of experimentally-induced tularemia in mice, using the live vaccine strain of Francisella tularensis . A sublethal, intravenous inoculation of this organism caused in C57BL/6 strain mice an acute infection which lasted approximately 12 days . The clearance of Francisella from the bloodstream was shown to be complete by 5.5 hours postinfection . At this time, approximately twice as many bacteria were isolated from the spleen as from the liver . Mice which had recovered from a primary infection demonstrated a significant resistance to re-infection with autologous Francisella, a memory which persisted for at least 15 weeks . Resistance to experimental tularemia could be passively transferred from infected mice to naive mice by means of non-adherent spleen cells . Cells capable of adoptive transfer of resistance were present at a maximal concentration 7 days following infection, and persisted in significant numbers within the spleen cell population for at least 20 days after infection . Treatment of mice with serum from recovered animals caused a decrease in resistance when measured in the livers, and an increase in resistance when measured in the spleens . Suppression of T cell-mediated immunity during infection by treatment with cyclosporin A resulted in a dramatic increase in the tissue bacterial counts . Cyclosporin A-induced suppression of antitularemic resistance was first noted 2-3 days following infection and remained apparent for at least 8 days . The results of these experiments demonstrate that resistance to experimental murine tularemia is mediated predominantly by a cell-mediated mechanism . This mechanism involves T cells which become activated as early as 2-3 days following infection . Experimental, non-lethal infection with Francisella tularensis is thus an excellent model for investigating the mechanisms of acquired cellular immunity. Biomed Chromatogr, 1987, 2(2), 76 - 8 Evaluation of vaginal malodor and efficacy of treatment by high performance ion exchange chromatography; Stanek R et al.; High performance ion exchange chromatography was employed to evaluate the presence of short chain organic acids in the vaginal fluid of a woman troubled by persistent foul vaginal odor, but who did not have typical bacterial vaginosis . The vaginal secretions from this patient were collected on a weighed cotton swab and eluted into water and extracted by acidified ether . Salts of the acids were back-extracted into aqueous solution and chromatographed on an H-form resin column and compared to commercially available standards . A strikingly large amount of caproic acid was found . The caproic acid disappeared after metronidazole therapy, and a subsequent follow-up chromatogram showed a predominance of lactic acid . The success of this technique in evaluating the present case suggests that such a method may prove useful in other types of vaginal infection. Arch Immunol Ther Exp (Warsz), 1987, 35(3), 289 - 302 Immunostimulation in recurrent respiratory tract infections therapy in children; Prusek W et al.; Selected immunologic parameters and effectivity of immunotherapy was evaluated in 117 children (12-month-10-year-1-old) suffering from recurrent respiratory tract infections . All the children displayed a profound depression of T lymphocytes number, which resemble the situation seen in AIDS patients . An increase of serum IgM concentration was also noted . Immunotherapy included treatment with the following preparations: TFX and Levamisol which stimulate T cell functions, Broncho-Vaxom which stimulates specific antibody production and a complex herb preparation PADMA showing undefined general stimulatory activity . Separate group of children was subjected to climatotherapy in Czerniawa Sanatorium and received no immunostimulants . All methods of treatment employed had beneficial effect . The highest percentage of positive results was obtained in children receiving TFX and Levamisol . In all groups under study, an elevation of T cells percentage was observed . This was especially evident in Levamisol treated patients . There was no correlation, however, between T cells number and clinical improvement. Arch Oral Biol, 1987, 32(11), 765 - 72 Biochemical and scanning electron microscope study of lipids chloroform-methanol extracted from unerupted and erupted human tooth enamel; Goldberg M et al.; Almost twice as much fatty acid was extracted without demineralization from the mature enamel of erupted teeth treated with 2:1 chloroform-methanol as from the enamel of unerupted teeth (32.64 micrograms/g versus 15.60 micrograms/g) . Palmitic C16:0, stearic C18:0, oleic C18:1 omega 9 and linoleic acids C18:2 omega 6 were the chief fatty acids detected by gas-liquid chromatography . The percentage of free fatty acids and diglycerides was larger in unerupted than erupted teeth, whereas that of triglycerides, phosphatidylethanolamine and phosphatidylcholine was larger in erupted teeth . These changes may be due to bacteria and other contaminants (saliva and pellicle) . The large amounts of free fatty acids, diglycerides, cholesterol and phospholipids as intrinsic components were probably due to the persistence of membrane remnants entombed during enamel formation, as indicated by the visualization of holes and by the increase in the size and number of focal holes after lipid-solvent interaction with the enamel surface . These defects always occurred in rods and never in inter-rod material . Other minute defects, including empty-widened intercrystal spaces, were also observed inside some rods. Acta Med Austriaca, 1987, 14(1), 1 - 4 {Clinical manifestations of acquired immunologic deficiency syndrome (AIDS)}; Goebel FD; The acquired immunodeficiency syndrome AIDS is caused by the retrovirus HIV . About 20% develop after the inoculation of the virus an acute clinical picture resembling infectious mononucleosis . Several weeks to months after the infection antibodies can be demonstrated in the serum . Lateron a lymphadenopathy syndrome or AIDS related complex may develop . Most of the patients with LAS or ARC will progress to the full blown picture of AIDS . This is defined as immunodeficiency complicated by Kaposi-sarcoma or central nervous system malignance lymphoma or opportunistic infections . The most common infections are due to certain parasites, c . e . pneumocystis carinii, toxoplasma gondii and cryptosporidia . Fungi, bacteria and viruses can also cause opportunistic infections. Dev Biol Stand, 1987, 67, 171 - 6 Recombinant DNA manipulations and their interfaces with protein technology; Nagley P; In recent years molecular biologists have learnt to mimic and exploit the natural processes of cell biology which are based on the fundamental principle that the amino acid sequence of a protein is encoded within the nucleotide sequence of DNA in its gene . The resultant set of novel methodologies, termed recombinant DNA technology, or genetic engineering, has revolutionized the study of almost all branches of biology and has presented radical new methods of producing proteins otherwise available only in very small quantities . Central technical features of the recombinant DNA technology are the ability to isolate and propagate individual genes from natural sources, and to express into the protein products the information intrinsic to such genes, using the most appropriate biological production systems (e.g . bacteria, yeast or cultured mammalian cells) . Embracing powerful analytical techniques such as DNA sequencing, and novel gene manipulations based on chemically synthesized oligonucleotides, the recombinant DNA technology is now having substantial impact on analytical and preparative protein technologies . The contemporary integrated approach to protein engineering involves not only protein chemistry and tools such as antibodies, but also has a dynamic interactive contribution from the recombinant DNA technology and its gene manipulation and expression skills . This integrated approach is particularly relevant to blood components such as clotting factors, antibodies, hormones and other proteins or peptides of powerful biological activity and specificity . Current work is focussed on the large-scale production of individual proteins as well as the molecular dissection of their biological actions and the development of products displaying novel biological activity. Proc Natl Acad Sci U S A, 1987 Jan, 84(1), 6 - 10 Cloning and cDNA sequence of the regulatory subunit of cAMP-dependent protein kinase from Dictyostelium discoideum; Mutzel R et al.; cDNA clones encoding the regulatory subunit of the cAMP-dependent protein kinase (ATP:protein phosphotransferase, EC 2.7.1.37) from Dictyostelium discoideum were isolated by immunoscreening of a cDNA library constructed in the expression vector lambda gt11 . High-affinity cAMP-binding activity was detected in extracts from bacteria lysogenized with these clones . Nucleotide sequence analysis of three overlapping clones allowed the determination of a 1195-base-pair cDNA sequence coding for the entire regulatory subunit and containing nontranslated 5' and 3' sequences . The open reading frame codes for a protein of 327 amino acids, with molecular weight 36,794 . The regulatory subunit from Dictyostelium shares a high degree of homology with its mammalian counterparts, but is lacking the NH2-terminal domain required for the association of regulatory subunits into dimers in other eukaryotes . On the basis of the comparison of the regulatory subunits from Dictyostelium, yeast, and bovine tissues, a model for the evolution of these proteins is proposed. Ann Rech Vet, 1987, 18(4), 429 - 37 Anti-Brucella cell-mediated immunity in mice vaccinated with a cell-wall fraction; Plommet M et al.; Immunity against Brucella, a facultative intracellular bacteria, can be induced in mice by live or by killed-whole-cell or fraction vaccine . This immunity can be adoptively transferred from live vaccinated donor to recipient mice, or passively with immune serum raised against bacterial fractions . A protein-bound peptidoglycan fraction (PG) was used to immunize DBA/2 donor mice intravenously or subcutaneously in the hind footpads . Splenic or popliteal lymph nodes cells were transferred to recipients that were intravenously challenged with a B abortus virulent strain . Brucella spleen counts, and in some cases liver counts, were done 15 days later or at successive times . Spleen cells depleted from macrophages and lymph nodes cells conferred immunity to recipients . This immunity increased after transfer for about 21 days . Lymph nodes cells were more efficient to transfer immunity than spleen cells . This immunity was abated but not abrogated by treatment of cells by anti-Thy serum and was transferred by both nylon wool non adherent (T enriched) or adherent (B enriched) cells . Comparative transfer experiments with DBA/2 versus CBA mice, live vaccine versus PG fraction vaccine, virulent versus avirulent (vaccinal) challenge evidenced that the three factors were involved in induction and/or expression of immunity . DBA/2 mice responded better to fraction vaccine and virulent challenge whereas CBA responded better to live vaccine and to avirulent challenge strain . In contrast DBA/2 X CBA F1 responded equally well against the virulent challenge to transfer of cells from live vaccine or PG fraction vaccinated donor mice . Two cell-mediated immune mechanisms may thus be involved in transferred immunity the expression of which may be genetically determined. Pediatrie, 1987, 42(8), 585 - 7 {Treatment of purulent leukorrhea in young girls}; Berlier P; The case of a prepuberal girl complaining of a more or less purulent genital discharge, eventually associated with bleeding, evokes a vulvovaginitis . Vulvovaginitis is much less frequent than vulvitis . When a vulvovaginitis is resistant to medical treatment or is recurrent, an endoscopic examination is required in order to eliminate the presence of a vaginal foreign body . Treatment includes local cleaning, local antibiotics and sometimes a short course of oestrogen therapy . Oral antibiotics are indicated only in specific germ infections. Pediatr Pathol, 1987, 7(4), 447 - 55 Mineral oil embolization and lipid pneumonia in an infant treated for Hirschsprung's disease; Rabah R et al.; A 5-month-old infant girl with Hirschsprung's disease died 1 month after colostomy as a result of mineral oil embolism and lipid pneumonia . She had received multiple mineral oil enemas and irrigations as treatment for impacted stools . Mineral oil peritonitis was present on the surface of the bowel adjacent to the stoma and covered large portions of the surface of the liver and spleen . There was 600 cc of slightly cloudy and bloody peritoneal fluid . The peritoneal exudate contained bacteria that were associated with only a minimal inflammatory reaction . This case calls attention to a previously unreported complication of the use of mineral oil. Microbiol Immunol, 1987, 31(11), 1113 - 5 A plasmid of group Q which confers resistance to trimethoprim and sulfonamides; Hedges RW; A 5.2-Mdal plasmid, determining resistance to trimethoprim and sulfonamides, is a member of incompatibility group Q. Arkh Patol, 1987, 49(11), 16 - 26 {Diagnosis of infectious diseases in the practice of pathologic anatomists}; Tsinzerling AV; A review of laboratory and morphologic methods currently used to study tissues in diseases caused by viruses, bacteria, mycoplasmas, fungi, and protozoa . Special attention is given to cytologic examination of smears and scrapings from viscera whereby valuable information that should be included in the death certificate can be obtained before histologic sections have been prepared and the laboratory examination has been completed. Am J Ind Med, 1987, 12(6), 759 - 64 The health investigation of cotton textile workers in Beijing; Liu MZ; This study reports findings from a survey of byssinosis among 289 cotton workers in certain cotton textile mills in Beijing . Incidence of byssinosis was 4.2% (12 cases) . It decreased with a lower concentration of cotton dust and increased with yearly increment of cotton dust exposure . An elevation in body temperature over 37 degrees C was present among 14.2% of the workers on the first working day after rest . Acute lung function decrement was related to high dust concentration in the rooms . The higher the dust concentration in the rooms, the more evident was the chronic lung function decrement . Chronic bronchitis in cotton workers was higher than in controls . Incidence of byssinosis in smoking cotton workers was higher than in nonsmoking cotton workers. Acta Neuropathol (Berl), 1987, 75(2), 185 - 98 Brain pathology induced by infection with the human immunodeficiency virus (HIV) . A histological, immunocytochemical, and electron microscopical study of 100 autopsy cases; Budka H et al.; Neuropathological examination of brain tissue of 100 patients with infection by the human immunodeficiency virus (HIV), including 98 with clinically manifest acquired immune deficiency syndrome (AIDS), revealed distinct multifocal-disseminated and diffuse brain tissue lesions, which can be regarded as HIV-induced brain lesions: multifocal giant cell encephalitis (MGCE; 4) and progressive diffuse leukoencephalopathy (PDL; 25) . These lesions were found in 38 brains, and in 17 in absence of infectious, necrotizing or inflammatory changes of other types . In 13 brains, a combination of MGCE with PDL was seen, suggesting a spectrum of HIV-induced brain lesions . MGCE is characterized by perivascular accumulations predominantly of rod cells, monohistiocytes and macrophages, all of which are strongly labeled with a monoclonal antibody to macrophages . Most conspicuous are multinucleated giant cells which are also labeled by anti-macrophage antibody, and which can be regarded as evidence of the local presence of HIV, as confirmed by electron microscopical detection of HIV particles in four MGCE brains, and by immunocytochemical detection of HIV proteins in two MGCE brains . PDL is characterized by a triad: diffuse myelin loss, astroglial proliferation, and infiltration by mono- and multinucleated macrophages . HIV-induced lesions can be morphologically differentiated from histopathological brain lesions known in immunosuppression, including what is called here nodular encephalitis {"subacute encephalitis" of the literature, in most cases attributable to cytomegalovirus (CMV) or toxoplasmosis}, by their characteristic histopathology including the hallmark presence of multinucleated giant cells, by direct immunocytochemical and electron microscopical demonstration of HIV in the lesions, and by the absence of opportunistic agents (bacteria, fungi, Toxoplasma, CMV, HSV or papovaviruses) . Diffuse poliodystrophy (diffuse proliferation of astroglia with swollen nuclei, occasionally minor neuronal loss and rod cell proliferation) was found in the cerebral cortex and other gray matter in half of all brains, including cases with gyral atrophy, and may be another correlate of HIV damage to the brain . Morphological delineation of HIV-induced brain lesions is a necessary prerequisite for a meaningful clinical definition of HIV-induced cerebral disease. J Math Biol, 1987, 25(5), 543 - 51 A piecewise-linear growth model: comparison with competing forms in batch culture; Kolker Y; A simplified one-dimensional growth model based on the mass conservation law is proposed . Mathematically, it represents an easiest special case of so called L-systems . The model developed being considered as a descriptive tool of the growth curves is compared with four other similar and widely used models, in application to numerical data of bacteria (taken from literature) and of algae (an original experiment) growing in batch . Identification of free constants of the five models in comparison is performed using a nonlinear least squares iterative procedure, for which special programs were written . Theoretical and practical (approximative) merits of the new model are shown and discussed. Biorheology, 1987, 24(6), 541 - 9 Secretion and ion transport in airways during inflammation; Nadel JA et al.; Water and secretions interact in airways to produce the sol and gel layers that allow for entrapment of foreign materials and subsequent clearance by ciliary movement and by cough . Active Cl ion transport produces fluid, and this process is activated by products of mast cells (leukotrienes), eosinophils (major basic protein), and by other inflammatory mediators (prostaglandins, bradykinin) . Gland secretions produce the bulk of the volume of secretions . Airway irritation stimulates gland secretion reflexly via vagal muscarinic pathways . Recently, the sensory nerves have been discovered to release substance P and other neuropeptides when the airways are irritated . The stimulatory effects of neuropeptides on gland secretion (and on other inflammatory sites) are modulated by enkephalinase a membrane-bound enzyme that cleaves neuropeptides and thereby inactivates them . Up- or down-regulation of enkephalinase is predicted to change the degree of inflammatory response to neuropeptides . Finally, the cell surface of airway epithelial cells have been discovered to secrete large molecular weight glycoconjugates; these secreted products are increased markedly by a series of proteinases produced by inflammatory cells (neutrophils, mast cells) and by bacteria . Their exact physiologic roles are still unknown but they may contribute to the bulk and viscoelastic properties of airway secretions, and they may serve an important role in bacterial, viral and inflammatory cell adhesion. Ciba Found Symp, 1987, 131, 124 - 39 Relationship of tumour necrosis factor and endotoxin to macrophage cytotoxicity, haemorrhagic necrosis and lethal shock; Rothstein JL et al.; In this communication we discuss preliminary evidence suggesting a very strong synergism between tumour necrosis factor (TNF) and lipopolysaccharide (LPS) or between TNF and other bacteria in causing haemorrhagic necrosis and lethal shock . We found that TNF by itself does not cause haemorrhagic necrosis when injected into normal skin . TNF also had a rather low systemic toxicity when injected into disease-free, germfree-derived, defined-flora animals . On the other hand the addition of small amounts of LPS markedly raised the lethality of intravenous TNF treatments, and LPS injected into normal skin 'prepared' the site of injection for subsequent induction of haemorrhagic necrosis by locally injected TNF . Similar synergism was observed between TNF and mycoplasma . We suggest that the synergism between TNF and bacterial endotoxin (or other bacteria or bacterial products) may be part of an important defence mechanism against infections which is independent of specific immunity mediated by B and T cells . This synergism may be useful in increasing the therapeutic effects of TNF on tumours if the development of systemic toxicity in this treatment can be prevented. Cancer Invest, 1987, 5(6), 615 - 25 Perioperative blood transfusion and solid tumor recurrence--a review; Blumberg N et al.; Evidence regarding the association of blood transfusions with recurrence of solid tumors is largely conflicting . This is perhaps not surprising given the retrospective nature of the studies, the complexity of the disease and its treatment, and variations in local transfusion practices . Nonetheless, data demonstrating that transfusions of whole blood are associated with earlier cancer recurrence are most readily explained by a cause and effect relationship . There is a growing literature documenting previously unforeseen immunologic consequences of homologous blood transfusion . These possible clinical consequences may include earlier cancer recurrence, and increased susceptibility to infection with bacteria and viruses . The questions raised in this review will likely be answered by further studies . For the present, the prudent clinician will select red blood cells rather than whole blood for transfusion, and recognize that blood transfusion is a therapy with considerable benefits, but also considerable risks. Dev Comp Immunol, 1987 Fall, 11(4), 679 - 704 A review of fixed phagocytic and pinocytotic cells of decapod crustaceans, with remarks on hemocytes; Johnson PT; Clearance of foreign materials from the hemocoel of decapod crustaceans involves several distinct kinds of cells . Proteins, and possibly viruses below 30 nm diameter, are known to be removed by the branchial podocytes, which are specialized pinocytotic nephrocytes located in the gills . These cells, and podocytes of the antennal gland (excretory organ) which may also be of importance in clearance, are similar to podocytes of Bowman's capsule of the vertebrate kidney . Phagocytic clearance of particulate material (with possible exception of the small viruses) is accomplished by three kinds of cells, one free and the other two fixed to tissues facing hemal spaces . The hemocytes, free cells suspended in the hemolymph, are the most abundant and generalized of these phagocytic cells . Phagocytic reserve cells are applied to myofibers in the heart of penaeid and palaemonid shrimp, and probably the same as reserve cells found throughout the hemocoel of all decapods . They are minimally involved in clearance of injected carbon and carmine, but are not known to phagocytize naturally occurring particles such as bacteria or viruses . The fixed phagocytes are very important in the clearance of some substances . They are located on the exterior surfaces of arterioles in hemal spaces of the hepatopancreas . They are highly specialized cells designed to sequester large amounts of various types of particulate matter, not only through phagocytosis but by trapping and retaining vast numbers of particles within a net- or sievelike layer of granular material which surrounds the free surface of the fixed phagocyte . Derivation, morphology, development, and function of the above cell types are described and discussed. Am J Ind Med, 1987, 12(6), 737 - 42 Some opinions on byssinosis in China; Xing GC; On the basis of some study results in China and reports from abroad, a conclusion should be drawn that leads to the recognition of a pneumoconiosis-like lesion of interstitial fibrosis . This disease is called "cotton pneumoconiosis" and we may classify these occupational lesions into four types: 1) byssinosis; 2) chronic obstructive pulmonary disease (COPD); 3) cotton fever; and 4) cotton pneumoconiosis . Byssinosis, COPD, cotton fever and cotton pneumoconiosis may be different types of responses due to the different duration of exposure, the different parts of bronchial tree (upper respiratory tract, small airway, and respiratory part) where deposition occurs, and the different components of cotton dust (broken cotton fibers, bracts, pericarps, bacteria, and fungi) . These responses, which include histaminelike reaction, allergy, and stimulation of foreign material, happen in different symptoms of the syndrome among cotton mill workers . But no matter whether responses caused by inhalation of dust are inflammation or allergic reaction, cotton dust is foreign stimulation on deposited sites, causing lung fibrosis after lung stimulation . For the health of cotton workers, we must pay attention not only to the acute effects but also to the chronic lesions . We therefore suggest that these four types of occupational lung disorders caused by inhalation of cotton dust may be called by the joint name, "Byssinosis syndrome." Arch Oral Biol, 1987, 32(7), 473 - 6 Proteolytic activities in the supragingival plaque of monkeys (Macaca fascicularis); Smith K et al.; Thirteen peptidase substrates were hydrolysed by this plaque . After fasting the monkeys for 24 h, the rates of hydrolysis of N-leucyl-, N-alanyl-, N-isoleucyl- and N-tyrosyl-2-naphthylamines, glycyl-L-proline 4-nitroanilide, N alpha-benzoyl-DL-arginine-2-naphthylamine and L-leucyglycine were significantly increased . Changes in these peptidase activities may serve to enhance the mobilization of amino acids and peptides, and they may explain why the growth rates of dental-plaque bacteria in vivo are generally unaffected by the availability of the host's diet. J Dent Res, 1987 Jan, 66(1), 2 - 9 Possible mechanisms involved in the immunoregulation of chronic inflammatory periodontal disease; Seymour GJ; It is generally agreed that immunological mechanisms are involved in the pathogenesis of periodontal disease; however, regulation of these mechanisms has hitherto received scant attention . Regulatory networks exist at both a cellular and a molecular level . At the cellular level, the existence of helper (T4-positive) and suppressor (T8-positive) T lymphocytes, the expression of Class II major histocompatibility complex antigens, and the heterogeneity of macrophage subpopulations are central to an understanding of the regulatory mechanisms involved . It is only recently that studies of these separate components, in both humans and experimental animals, have begun to provide a basis for understanding the complex interactions occurring in periodontal disease . Studies using the human experimental gingivitis model have shown an immunoregulatory picture consistent with a controlled immunological reaction with an essentially normal T4:T8 ratio of 2.0 . In contrast, studies utilizing cells extracted from adult periodontitis lesions have shown a reduced T4:T8 ratio (approximately 1.0) and an inability to respond in, or to stimulate, an autologous mixed lymphocyte reaction . Animal studies using athymic nude rats have supported the concept of a central role for T-cell control in periodontal disease and the possibility of an imbalance in this control with disease progression . These results are reviewed and areas of future research explored. Annu Rev Nutr, 1987, 7, 157 - 85 Dietary regulation of gene expression: enzymes involved in carbohydrate and lipid metabolism; Goodridge AG; The mechanisms of the responses of an enzyme to different hormones and metabolites or several enzymes to a single hormone are surprisingly varied . There is neither an operon for lipogenic enzymes nor a common step at which hormones and metabolites coordinately regulate the expression of lipogenic genes . In bacteria, coordinated expression of several enzymes in a single metabolic pathway often is achieved by organizing the genes into operons . An operon is a group of genes linked together in a linear fashion and producing a polycistronic mRNA . Trans-acting factors regulate the transcription of these genes by interacting with promoter/regulatory sequences in the 5'-flanking region of the most 5'-ward of the genes . In vertebrate animals, however, coordinated control of gene transcription is not achieved by linking the individual genes, but by putting in the 5'-flanking regions of these genes a regulatory sequence that interacts with common trans-acting factors . Genes controlled by different hormones are expected to have regulatory elements for each hormone . The presence of glucocorticoid and cyclic AMP regulatory elements at the 5'-end of the PEPCK gene is consistent with this notion . Transcription is not the only step at which hormones and metabolites control the pathways for gene expression . The levels of the mRNAs for L-PK, ME, S11, and S14 are increased by T3 at post-transcriptional steps . Glucagon also regulates the accumulation of ME mRNA post-transcriptionally . Neither the mechanism nor the sequence organization of regulatory elements is known for post-transcriptional control of gene expression . In the case of PEPCK and HMG-CoA reductase, the next steps will be to determine more precisely the sequences in the 5'-region that mediate hormone sensitivity and feedback inhibition, respectively, and whether trans-acting factors are involved . For the other genes discussed, identification of the regulated step must precede identification of sequences that confer hormone or metabolite-sensitive regulation on a specific gene . In general, it is probable that the hybrid gene approach, so successful for PEPCK and HMG-CoA reductase, also will be effective in defining cis-acting hormone- or metabolite-regulatory elements in other genes . These techniques should be applicable to both transcriptional and post-transcriptional mechanisms . Our long-term objective is to understand the molecular basis of each event that intervenes between the binding of hormone or metabolite to its appropriate receptor and altered enzyme level.(ABSTRACT TRUNCATED AT 400 WORDS) Comp Biochem Physiol B, 1987, 86(3), 423 - 38 What is the function of protein carboxyl methylation? van Waarde A. The following functions of protein carboxyl methylation seem to be reasonably well established: Multiple, stoichiometric methylation of chemotactic receptors in bacteria at glutamyl residues serves as one (but not the only) adaptation mechanism of the transduction chain to constant background levels of chemotactic stimuli . Stoichiometric methylation of hormones and hormone carrier proteins plays a role in hormone storage and secretion by the pituitary gland . Substoichiometric methylation at D-aspartyl residues is involved in a repair mechanism of aged proteins . Stoichiometric methylation of calmodulin modulates the sensitivity of calmodulin-dependent processes to calcium . Research of the past 3 years has indicated that in order to demonstrate an involvement of methylation in the coupling of surface receptors to intracellular events three new criteria have to be met: (a) the cell should possess a protein carboxyl methylase with relatively narrow substrate specificity; (b) methylation should take place at L-amino acid residues; (c) the methyl accepting proteins should be methylated in a stoichiometric fashion. Eur J Cardiothorac Surg, 1987, 1(3), 158 - 64 Factors predisposing to wound infection in cardiac surgery . A prospective study of 517 patients; Wilson AP et al.; Postoperative wound infection can greatly prolong hospital stay after cardiac surgery, so the identification of predisposing factors may help in prevention or early institution of treatment . Transfer of organisms from the leg to the sternum during coronary artery surgery has been proposed as a major additional cause of sepsis . The definition of wound infection is not standardised and therefore makes comparison between centres difficult . In a prospective study of 517 patients, a wound scoring method (ASEPSIS) has been used to register all abnormal wounds to maximise the chances of identifying factors predisposing to infection . Abnormal healing was noted in 99 (19%) sternal wounds and 29 (8%) leg wounds . Obesity was the principal risk factor (P less than 0.005) . Diabetes, reoperation, length of preoperative hospital stay, age, sex, or previous cardiac surgery had little effect on wound healing . The range of bacteria isolated from chest wounds after coronary artery surgery was similar to that after valvular surgery, but the rate of isolation was significantly greater . With careful attention to technique, leg wound infection rarely presented a clinical problem and did not appear to be a source of bacteria infecting the chest wound. Pol Arch Weter, 1987, 27(1), 81 - 91 {Activity of porcine anti-Brucella abortus immunoglobulins in the acid plate agglutination test (APAT)}; Stryszak A et al.; Serological activity of swine IgM and IgG against Brucella abortus in RBPT was determined in relation to four other reactions used in Poland for diagnosing brucellosis standard agglutination test, complement fixation test, antiglobulin test, 2-mercaptoethanol test) . Isolation of IgG was performed by the method of filtration on Sephadex gel G-200 of swine sera raised against Brucella abortus S19 by double immunization with suspension of killed bacteria . The presence of a certain Ig class in the fractions thus obtained was confirmed by immunoelectrophoresis and immunodiffusion tests . RBPT revealed the reaction of antibodies of IgM and IgG class which proves usability of this reaction diagnosis both early (IgM) and chronic (IgG) infection with brucellosis . Both classes of antibodies mentioned above were active also in SAT and CTT . Also the results obtained in AGT and MET were found interesting . In one of the sera, the absence of incomplete antibodies was observed, whereas positive reaction in antiglobulin test was found in its fractions containing IgG . This phenomenon was determined as concealment of incomplete agglutinins through higher level of complete antibodies in normal serum . In swine (the results were different from those obtained for cattle), apart from incomplete antibodies in IgG class, the presence of these agglutinins in IgM class was noted . On the other hand, the results obtained in MET proved that IgM antibodies of swine were not totally reduced when affected by 2-mercaptoethanol. Gene, 1987, 59(1), 77 - 86 Cloning of a Drosophila melanogaster adenine phosphoribosyltransferase structural gene and deduced amino acid sequence of the enzyme; Johnson DH et al.; The Aprt locus of Drosophila melanogaster encodes the structural gene for adenine phosphoribosyltransferase (APRT) . DNA cloned from microdissected salivary gland polytene chromosome region 62B7-12 was used in conjunction with chromosome walking and hybrid selection of mRNA to isolate the Aprt gene . Aprt lies at cytogenetic position 62B9 and is closely flanked by other genes of unknown function . Nucleotide sequencing shows that four APRT cDNAs have a common 5' terminus with an apparent cap consensus sequence but two different 3' sites of polyadenylation . The distribution of conserved amino acid sequences in APRT from vertebrates, insects and bacteria suggests that they may have shared a common ancestral gene for this ubiquitous enzyme. Lasers Surg Med, 1987, 7(6), 467 - 72 Endogenous porphyrin fluorescence in tumors; Harris DM et al.; The fluorescence of endogenous porphyrins in tumors has been attributed to the presence of several different species of bacteria inhabiting ulcerated and necrotic tissues . We have examined this "autofluorescence" in carcinogen-induced tumors in the hamster oral mucosa and in patients with cancer of the oral cavity . In several instances, bright autofluorescence of normal tissues has been observed . This represents a potential false-positive identification of cancer when systemic hematoporphyrin derivative fluorescence is used as a diagnostic technique. Acta Otolaryngol Suppl, 1987, 435, 85 - 9 Prognosis and endotoxin contents in middle ear effusions in cases after acute otitis media; Iino Y et al.; We analysed the endotoxin content of middle ear effusion (MEE) from patients after acute otitis media, by a Limulus assay . Endotoxin was positive in 70% of the cases treated with antibiotics at an acute event for more than 5 days, while the incidence of bacteria was only 22% . One month after the sample collection, cases with a high concentration of endotoxin in MEE still tended to have effusion . These results suggest that endotoxin in MEE cannot be easily inactivated and may be involved in the development of acute otitis media into chronic otitis media with effusions. Langenbecks Arch Chir, 1987, 370(3), 173 - 83 {Measuring chemiluminescence in phagocytic granulocytes--simultaneously a parameter of their killing function?}; Dziwisch L et al.; The results of the chemiluminescence activity determined in whole blood samples very often were used to compare the unspecific defense mechanism of polymorphonuclear granulocytes (PMN) against causative organisms to preoperative capable risk parameters in surgery . Our investigations try to compare the results of a bacteria-induced chemiluminescence with those we received from a standardized agar killing plate-test . It could be shown that there is no correlation between the used two test systems . Therefore it is not possible to conclude from the chemiluminescence results especially made out of whole blood samples to the potency of the defense mechanism of PMN granulocytes. J Mol Evol, 1987, 24(4), 357 - 65 Accumulation pattern of amino acid substitutions in protein evolution; Kunisawa T et al.; A simple method for the evolutionary analysis of amino acid sequence data is presented and used to examine whether the number of variable sites (NVS) of a protein is constant during its evolution . The NVSs for hemoglobin and for mitochondrial cytochrome c are each found to be almost constant, and the ratio between the NVSs is close to the ratio between the unit evolutionary periods . This indicates that the substitution rate per variable site is almost uniform for these proteins, as the neutral theory claims . An advantage of the present analysis is that it can be done without knowledge of paleontological divergence times and can be extended to bacterial proteins such as bacterial c-type cytochromes . It is suggested that the NVS of cytochrome c has been almost constant even over the long period (ca . 3.0 billion years) of bacterial evolution but that at least two different substitution rates are necessary to describe the accumulated changes in the sequence . This "two clock" interpretation is consistent with fossil evidence for the appearance times of photosynthetic bacteria and eukaryotes. Differentiation, 1987, 33(3), 197 - 206 Distinct developmental regulation and properties of the responsiveness of different genes to cyclic AMP in Dictyostelium discoideum; Bozzone DM et al.; Cyclic AMP (cAMP) is known to be an important mediator of gene expression in eukaryotic cells . At present, little is known about the developmental events which render specific genes responsive to cAMP in distinct cell types, or about the biochemical mechanisms by which cAMP exerts these regulatory effects . By examining the effects of cAMP treatment on specific mRNA levels in Dictyostelium discoideum cells with different 'developmental histories', we defined the developmental states in which specific genes display responsiveness to cAMP . We focused on two specific rapid responses: the ability of cAMP to inhibit the expression of an 'early' developmentally regulated mRNA (discoidin-I) and to stimulate the expression of a 'late', prespore-specific mRNA (PL3) . Using this approach, we showed that, for both mRNAs, the ability to respond rapidly to cAMP is absent from vegetative cells grown on bacteria, and is acquired during development on filters . Furthermore, we identified several developmental states in which the discoidin-I response to cAMP is present, but in which the PL3 response is not . In experiments designed to examine the effects of cAMP analogues on the levels of these two mRNAs, we demonstrated that the analogue specificities of the discoidin-I and PL3 responses are different, and that the specificity for the PL3 response depends on the developmental state . The developmental kinetics and analogue specificity of the PL3 response suggest a two-step mode of action of cAMP in activating the expression of this gene . We discuss possible implications of these findings for the mechanisms of action of exogenous cAMP as well as for the role of cAMP in controlling the changes in gene expression that accompany normal development. Parasitol Res, 1987, 73(3), 199 - 202 Detection of Leishmania parasites by DNA in situ hybridization with non-radioactive probes; van Eys GJ et al.; In situ hybridization techniques develop rapidly into diagnostic tools of considerable value for detection of viruses and bacteria . Here we report the application of this technique for the detection of Leishmania parasites . Biotin-labelled total promastigote DNA was hybridized to cultured Leishmania parasites and to blood and impression smears of infected mice . In promastigotes kinetoplasts were strongly stained, nuclei somewhat more diffuse . In amastigotes both nuclear and kinetoplast DNA hybridized strongly . Amastigotes were easily detected in tissue of infected mice by their stable configuration of kinetoplast and nuclei . Cross-hybridization was observed between Leishmania donovani and L . tropica, but not between these two and L . braziliensis or Trypanosoma cruzi . A minor aspecific staining of host cell nuclei in the smears did not interfere with the detectability of the parasites. J Cell Biochem, 1987 Jan, 33(1), 39 - 51 Site-specific mutagenesis of cDNA clones expressing a poliovirus proteinase; Semler BL et al.; The cleavage of poliovirus precursor polypeptides occurs at specific amino acid pairs that are recognized by viral proteinases . Most of the polio-specific cleavages occur at glutamine-glycine (Q-G) pairs that are recognized by the viral-encoded proteinase 3C (formerly called P3-7c) . In order to carry out a defined molecular genetic study of the enzymatic activity of protein 3C, we have made cDNA clones of the poliovirus genome . The cDNA region corresponding to protein 3C was inserted into an inducible bacterial expression vector . This recombinant plasmid (called pIN-III-C3-7c) utilizes the bacterial lipoprotein promoter to direct the synthesis of a precursor polypeptide that contains the amino acid sequence of protein 3C as well as the amino- and carboxy-terminal Q-G cleavage signals . These signals have been previously shown to allow autocatalytic production of protein 3C in bacteria transformed with plasmid pIN-III-C3-7c . We have taken advantage of the autocatalytic cleavage of 3C in a bacterial expression system to study the effects of site-specific mutagenesis on its proteolytic activity . One mutation that we have introduced into the cDNA region encoding 3C is a single amino acid insertion near the carboxy-terminal Q-G cleavage site . The mutant recombinant plasmid (designated pIN-III-C3-mu 10) directs the synthesis of a bacterial-polio precursor polypeptide that is like the wild-type construct (pIN-III-C3-7c) . However, unlike the wild-type precursor, the mutant precursor cannot undergo autocatalytic cleavage to generate the mature proteinase 3C . Rather, the precursor is able to carry out cleavage at the amino-terminal Q-G site but not at the carboxy-terminal site . Thus, we have generated an altered poliovirus proteinase that is still able to carry out at least part of its cleavage activities but is unable to be a suitable substrate for self-cleavage at its carboxy-terminal Q-G pair. Nature, 1987 Jan 1-7, 325(6099), 70 - 3 Bovine papillomavirus E2 trans-activating gene product binds to specific sites in papillomavirus DNA; Androphy EJ et al.; Enhancers are cis-acting elements that activate transcription in higher eukaryotes independently of their position or orientation relative to the promoter that they activate . The mechanisms by which enhancers activate transcription are poorly understood, in part because, with the exception of the glucocorticoid receptor, the proteins that directly interact with enhancers have not been purified, nor have the genes encoding them been cloned . The upstream regulatory region (URR) that immediately precedes the early genes of the bovine papillomavirus type 1 genome (BPV) has enhancer activity when it is activated by a trans-acting gene product of the BPV E2 open reading frame (ORF) (Fig . 1) . It is not known whether this enhancement represents a direct or indirect effect of E2 on the URR . We have used an E2 peptide expressed in bacteria and a DNA-protein complex immunoprecipitation assay to study E2-mediated enhancement of transcription by the URR . We show here that this peptide directly binds to four specific sites in the BPV URR, and to one site in the human papillomavirus (H)PV16 URR . All the binding sites contain a related sequence of nucleotides; a 23 base pair (bp) fragment containing this sequence can specifically prevent binding of the E2 protein to the BPV URR . The BPV E2-URR enhancer interaction may therefore represent a useful model system for studying the mechanism of transcriptional enhancement, as both an effector protein and its target enhancer can be purified and genetically manipulated. Annu Rev Cell Biol, 1987, 3, 143 - 78 Molecular aspects of B-lymphocyte activation; DeFranco AL; The activation of B lymphocytes from the resting stage to the proliferating stage and then to the fully differentiated antibody-secreting stage is a highly regulated and complicated process . B-cell activation can clearly proceed by a number of different routes, each promoted by different regulatory cells (macrophages, two types of helper T cells) and each dependent upon the properties of the relevant antigen molecules . In most cases the nature of the antigen may simply control the magnitude and/or duration of antigen receptor signaling . An antigen that by itself generates inefficient signaling may require additional signals from helper T cells to induce B-cell activation . In contrast, antigens derived from bacterial cell surface components, such as LPS, can directly activate B cells and macrophages . This vigorous, polyclonal responsiveness to certain bacterial components probably represents a specialized system to enhance antibody responses to bacteria, whereas helper T cell-dependent responses may be primarily useful in making antibodies against soluble proteins and viruses . Thus multiple pathways of B-cell activation probably are needed to generate adequate antibody responses against the variety of pathogenic micro-organisms encountered by vertebrates. Dev Comp Immunol, 1987 Summer, 11(3), 649 - 60 Analysis of suppressor factor in delayed immune responses of a bat, Pteropus giganteus; Chakravarty AK et al.; Bat spleen and mesenteric lymph node cell cultures treated with varying doses of LPS showed significant blastogenic and DNA synthetic responses between 72-96h . Peak responses were not different when the cell culture medium was supplemented either with autologous serum or heterologous serum indicating the absence of any significant suppressor factor in autologous bat serum . In contrast, blastogenesis and DNA synthesis peaks appeared early, at 48h, in lymphocytes depleted of suppressor T cells by pretreatment with cyclophosphamide . Direct antibody producing cells against SRBC were studied in normal spleen and mesenteric lymph node cell cultures . The peak PFC response was lacking even at 120h while CY pretreated bat lymphocytes, showed peak PFC responses at 96h . Thus the delayed immune response in bats seems to be a function of suppressor T cells but serum suppressor factor(s) possibly exerts no significant effect . The function of suppressor cells in bats in relation to their role as carriers of several dreaded bacteria and viruses is discussed. Isozymes Curr Top Biol Med Res, 1987, 16, 49 - 65 Modified beta-D-N-acetylhexosaminidase isozymes for enzyme replacement in GM2 gangliosidosis; Rattazzi MC et al.; The therapeutic potential of enzyme replacement in lysosomal storage disorders has remained largely unfulfilled, perhaps because of negative reactions to the initial disappointing results . Despite the existence of several animal models that can be utilized to explore solutions to the problems of exogenous enzyme targeting, the interest in ERT prevalent during the 1970's seems to have subsided to be replaced by active interest in bone marrow transplantation (BMT, Krivit and Paul {1986}) . This is a logical approach to enzyme replacement in storage disorders of the RE system, and indeed some encouraging results have been obtained . However, in addition to having high morbidity and mortality, in the ultimate analysis BMT presents the same targeting problems as conventional ERT . In our opinion, these problems can be solved more easily in the case of ERT by exploiting the existing cellular uptake mechanisms and infusing enzymes whose structure has been suitably modified by simple biochemical manipulations . Accordingly, we have explored a methodology that takes advantage of negative charges on the cell surface to obtain nonspecific but effective membrane binding of beta-hex coupled to the highly positively charged PLL, followed by internalization and routing to the lysosomes . This system increases uptake of exogenous enzyme by some neurons in vitro and possibly in vivo, but its efficiency depends on the cells' endocytic activity that, in the case of neuronal soma, apparently is low . Thus, we have chosen as recognition marker for specific neuronal uptake a nontoxic fragment of TTx that is efficiently taken up by these cells . The initial results are encouraging; they support our contention that effective enzyme replacement methodologies can be devised, and encourage us to continue our work in this direction . Finally, recombinant DNA techniques are now being applied to a number of LSD, and the genes for several of the pertinent enzymes have been or are being isolated . In addition to representing a first step towards gene replacement therapy, the results of this work will permit the generation of large amounts of human enzymes from bacteria by recombinant DNA methods, thus obviating the problem of enzyme supply for ERT . Since human lysosomal enzymes obtained from bacteria will be nonglycosylated, to obtain cell uptake it will be necessary to resort to the type of modifications that we are trying to develop at this time, i.e., covalent linkage to moieties that allow non-glycosyl-mediated cellular uptake . Thus, our work on beta-hex may provide a model for biochemical manipulations of bacterially produced enzymes applicable to several LSD. Comp Biochem Physiol B, 1987, 87(1), 35 - 9 Inhibition of energy metabolism by benzoxazolin-2-one; Niemeyer HM et al.; The effects of the title compound (BOA) on energy-linked reactions in mitochondria were studied . BOA inhibited electron transfer between the flavin and ubiquinone in Complex I, and ATP synthesis at the F1 moiety of the ATPase complex . These results are discussed in relation to the toxicity of BOA towards a wide range of aerobic organisms. Graefes Arch Clin Exp Ophthalmol, 1987, 225(1), 45 - 9 Immunological and immunopathological aspects of opsin-induced uveoretinitis; Broekhuyse RM et al.; In an extension of our previous studies, experimental autoimmune uveoretinitis (EAU) was induced in Lewis rats by injection of very high doses of bovine opsin . The induced reaction consisted predominantly of a mild posterior retinitis . Varying the amount of injected opsin between 300 and 1,000 micrograms did not influence this result, provided that the antigen was injected in Freund's complete adjuvant . Pathogenicity of opsin appeared to be lower than that of interphotoreceptor retinoid binding protein (IRBP) or S-antigen, while EAU induced by the latter antigens was much more dose-dependent than EAU induced by opsin . An increase of the dose strongly accelerated the onset and increased the incidence of EAU from low to moderate . However, severe inflammation and high incidence were only obtained by co-injection of Hemophilus pertussis bacteria . This adjuvant especially increased cellular immune responses to opsin as measured by lymphocyte transformation . No marked effects on humoral responses were detected by ELISA, using different types of opsin preparations . Development of opsin-induced EAU was inhibited by ciclosporin, a suppressor of certain specific T cell functions . Ciclosporin injections lowered the antibody response of the rats and eliminated measurable lymphocyte transformation in vitro . Induction of opsin-EAU therefore appears to be T-cell-dependent . The effect of pertussis adjuvant may be explained by enhancement of the T cell responses to opsin and by increasing the permeability of the blood-retina barriers . Other properties of the adjuvant may be of importance as well . A relationship between change in molecular conformation and uveitogenicity of opsin is discussed. Int Arch Allergy Appl Immunol, 1987, 82(2), 159 - 67 Effect of polymeric IgG on human monocyte functions; Mannhalter JW et al.; Fc and iC3b receptors are involved in various biological functions of phagocytic cells, such as immune adherence and phagocytosis of opsonized particles, degranulation and superoxide generation . In the present study we examined the expression of specific receptors for the Fc portion of IgG (FcR) and for iC3b (CR3), a cleavage product of the third complement component, on human monocytes following in vitro treatment with polymeric and monomeric IgG . Interaction of polymeric IgG (fluid phase) with the monocyte membrane led to a concomitant modulation of both Fc and iC3b receptors . Monomeric IgG, however, down modulated Fc receptor expression only if surface bound . Under these conditions, no concomitant modulation of the iC3b receptor could be observed . The down modulation of Fc and iC3b receptors induced by fluid-phase IgG polymers was also accompanied by a decrease in monocyte functions as expressed by reduced Fc receptor-mediated phagocytosis, decreased release of oxygen metabolites following stimulation by aggregated IgG and opsonized zymosan, as well as in impaired killing of bacteria . These data suggest that a down modulation of Fc and iC3b receptors might have important implications for host defense mechanisms, since interaction with these receptors is required for the proper elimination of many pathogens. Folia Microbiol (Praha), 1987, 32(5), 382 - 7 Characterization of the submerged growth of Moraxella bovis; Mandl M et al.; A submerged batch cultivation of Moraxella bovis in a medium containing enzymic casein hydrolyzate and supplemented with dialyzed ram blood was described . Up to the growth limitation the bacteria grew exponentially without a lag phase and with a doubling period of 64 min . During the exponential phase no significant decrease of viable cells and cell autolysis were observed . Amino acids were the limiting factor . At the end of growth glutamic acid, threonine and serine were detected at the lowest concentrations . Substrate limitation led to an irreversible decrease of the metabolic activity . Growth yield on oxygen was 2.3 X 10(9) cells per mg O2 . The fraction of respiration required for growth-uncoupled processes was negligible . During the cultivation ammonia was produced and the pH increased but it was not the inhibitory factor . Respiration was not limited by oxygen at concentrations higher than 0.63 mumol O2 per L . Sufficient pili were produced. J Biol Stand, 1987 Jan, 15(1), 17 - 26 The detoxification of Bordetella pertussis with glutaraldehyde; Iida T et al.; To improve the whole-cell pertussis vaccine we have studied the inactivation of the biological properties characteristic of Bordetella pertussis phase I bacteria, i.e . histamine-sensitizing, lymphocytosis-promoting and mouse protective activities, by treating a concentrated bacterial suspension with various concentrations of glutaraldehyde . Under the experimental conditions, treatment with 10 mM glutaraldehyde at 37 degrees C for 30 min resulted in a marked reduction of the toxic activities without grossly diminishing the protective potency . Further tests were performed on the stability of the protective potency, on the agglutinin production in mice, and on the freedom from abnormal toxicity in guinea-pigs. Am J Cardiovasc Pathol, 1987, 1(2), 147 - 56 Cardiovascular effects of congenital infections; Rosenberg HS; In the course of gestation, many bacteria, parasites, and viruses may infect the pregnant woman, but few cross the placenta to affect the fetus and fewer still affect the fetal heart . Although the incidence of fetal cardiac infection is low, the effect on the fetus is major . In terms of frequency, rubella virus, Toxoplasma gondii, and Coxsackie virus B are the principal infectious agents affecting the fetal heart, but any number of organisms may cross the placenta to affect the fetus . The pathogenesis of infection of the fetal heart relates to the agent and to the time of gestation when the infection occurs . The agent affects the heart along one or more of three separate pathways: inhibition of cell growth, cytolysis, and interference with the blood supply . Most agents cause cytolysis, stimulating inflammation and scarring . Although several agents carry the suspicion of teratogenicity, only rubella virus has been incriminated with certainty as capable of functioning along each of the three pathways with the potential to serve as teratogen. Curr Genet, 1987, 11(4), 309 - 14 Phylogenetic implication of heterogeneity of the nontranscribed spacer of rDNA repeating unit in various Neurospora and related fungal species; Verma M et al.; Nontranscribed spacer (NTS) regions of ribosomal (r)RNA genes are non-conserved and are shown to be useful for phylogenetic studies . 32P-labelled N . crassa NTS pCC3400 DNA, was used as a molecular probe to hybridize Southern blots of genomic DNAs obtained from Neurospora, Gelacinospora, Sordaria, bacteria, plants, and animals . Our studies conclude that: (a) the homotahllic species of Neurospora should not belong to genus Gelacinospora (a historical question) and that Neurospora homothallic species are closer to Gelacinospora than to Sordaria; and that (b) all of the filamentous fungal species tested are indeed closer to the higher plant genome than to higher primate animal genome based on shared restriction sites of 12 enzymes . Our studies also demonstrate the usefulness of nontranscribed rRNA gene probes in resolving questions regarding phylogenetic relatedness between widely separated organisms using the parsimony principle based on mutation sites from DNA restriction maps; it has not been possible to do this using DNA: DNA hybridization procedures that involved the total genome. Crit Rev Microbiol, 1987, 15(2), 117 - 40 The genetics and biochemistry of mercury resistance; Foster TJ; The ability of bacteria to detoxify mercurial compounds by reduction and volatilization is conferred by mer genes, which are usually plasmid located . The narrow spectrum (Hg2+ detoxifying) Tn501 and R100 determinants have been subjected to molecular genetic and DNA sequence analysis . Biochemical studies on the flavoprotein mercuric reductase have elucidated the mechanism of reduction of Hg2+ to Hg0 . The mer genes have been mapped and sequenced and their protein products studied in minicells . Based on the deduced amino acid sequences, these proteins have been assigned a role in a mechanistic scheme for mercury flux in resistant bacteria . The mer genes are inducible, with regulatory control being exerted at the transcriptional level both positively and negatively . Attention is now focusing on broad-spectrum resistance involving detoxification of organomercurials by an additional enzyme, organomercurial lyase . Lyase genes have recently been cloned and sequencing studies are in progress. Vet Med Nauki, 1987, 24(6), 10 - 8 {Etiological study of viral gastroenteritis in swine}; Ignatov G et al.; Studied were the etiology and spread of nonbacterial gastroenteritis in pigs over a period of five years . Ascertained were cases of diarrhea caused by the virus of transmissive gastroenteritis, epizootic diarrhea, and rotaviruses . The transmissive gastroenteritis virus was found in 14.1 per cent of the investigated material, mostly in the winter months (67.8 per cent), which was 65.6 per cent of the positive cases . The number of cases and that of the farms involved abruptly rose in the last two years of the investigated period when as many as 73.4 per cent of the positive findings were recorded . Thirty to 37.7 per cent of the studied farms proved to be infected . Enzootic diarrhea was recorded on seven out of the 12 studied farms, whereas 30 to 50 per cent of the sampled material was positive . The serologic study of reconvalescent blood sera of pigs from three infected farms revealed that 90 up to 100 per cent of them contained antibodies . In a great part of the sera (from 15 to 68.9 per cent) there were transmissive gastroenteritis antibodies as well . In 6.8 per cent of the Investigated samples and on 30 per cent of the farms there were also findings of rotaviruses . Cases of rotavirus gastroenteritis were recorded the year round with a rising trend in the cold months . Antibodies to these viruses were established on a large scale--from 58.13 to 100 per cent . There were also mixed infections--corona- and rotaviruses or viruses and bacteria. Boll Ist Sieroter Milan, 1987, 66(2), 158 - 60 {Antiviral activity of a chemical contraceptive (nonoxynol-9)}; Barbi M et al.; The aim of this study was to evaluate, in vitro, the antiviral activity of a chemical contraceptive (Nonoxynol-9) . The results have demonstrated that such a compound is able to inhibit the infectivity of enveloped viruses . It seems that the activity of this contraceptive is due to the lesion of the envelopePIP: The action in vitro of chemical intravaginal contraceptives against bacteria and fungi has already been demonstrated and its antiviral activity is now undergoing testing . Nonoxynol-9 was tested on herpes simplex type II, respiratory sincipital virus and polio virus type II; the viruses were grown in human fetal fibroblasts, continued-line human cells, and continued-line monkey kidney cells . A cytotoxic effect was demonstrated up to a 1-10 dilution of the basic solution within 1 minute, without variances between different cellular lines . The results were tested against untreated control colonies . This study shows the efficacy of 1 chemical contraceptive against unrelated viruses; the target seems to be the pericapsidic membrane . This suggests a possible role for these drugs in the prevention of sexually transmitted venereal diseases such as HTLV-III and herpes . Biochim Biophys Acta, 1987, 895(1), 25 - 39 Principles of coupling between electron transfer and proton translocation with special reference to proton-translocation mechanisms in cytochrome oxidase; Krab K et al.; The recent general acceptance of the proton-pumping function of cytochrome oxidase has stimulated discussion and experiment on possible underlying molecular mechanisms . Adequate experimental design requires clear understanding of the theoretical principles governing such a linked function . The increasing structural knowledge of cytochrome oxidase also contributes to a present-day requirement of more precise chemical and physical description of redox-linked proton translocation, which is the fundamental process underlying conservation of energy from aerobic metabolism in all eukaryotes and many bacteria . This essay is based on our original theoretical treatment of this problem, which is expanded here to include discussion of more recent analyses by others, classification of different types of coupling principles, as well as some concrete proposed molecular mechanisms . The latter will be analysed qualitatively, and in some cases quantitatively where this is possible, using a common theoretical framework to help comparison between models . Experimental findings relevant to this problem will be critically reviewed, and some suggestions will be made to stimulate further experiments dedicated to clarify the problem. Indian J Lepr, 1987 Jan-Mar, 59(1), 9 - 19 A rapid method for viability and drug sensitivity of Mycobacterium leprae cultured in macrophages and using fluorescein diacetate; Bhagria A et al.; The ability of viable M . leprae to hydrolyze Fluorescein diacetate and retain fluorescein inside the bacteria was used to identify viable M . leprae inside the cultured in vitro macrophages . The subjective microscopic count of the FDA test was demonstrated as useful routine test by confirming the results obtained therein with a quantitative and non subjective measurement of fluorescence in spectrofluorimeter . Using this method loss of viability of M . leprae in presence of dapsone and rifampicin was demonstrated . Such an assay, was well correlated with another in vitro assay, the Fc receptor test and also the in vivo mouse foot test . The drug resistance of clinical isolates of M . leprae demonstrated by mouse foot pad was also correlated with FDA test system . Thus we have reported a reliable, consistent and rapid in vitro test system for determining viability and drug sensitivity of M . leprae. Dev Biol Stand, 1987, 67, 281 - 7 The clinical use of intravenous gammaglobulin; Dwyer JM; The availability of safe and effective preparations of human immune globulin that can be administered intravenously has revolutionized replacement therapy for patients suffering from hypogammaglobulinaemia . Of equal importance and greater interest, however, has been the recognition that super physiological doses of IgG can manipulate an abnormal immune system . Future prospects for the use of immunoglobulin preparations to supply specific antibodies includes the standardization of procedures, whereby patients with acute sepsis may receive antibiotics and immunoglobulin simultaneously . Already there is in vitro evidence that suggests that opsonized bacteria are more readily affected by aminoglycosides . It seems certain that gamma globulin will be used routinely in the management of patients with a number of immunomalignancies, such as chronic lymphatic leukaemia and multiple myeloma that feature hypogammaglobulinaemia, especially when chemotherapy is being administered . Control trials are underway to determine whether gamma globulin given intravenously to premature babies will satisfactorily correct their immuno-deficient state and improve their chances of survival . The immunomanipulative capacity of immunoglobulin is yet to be fully realized . Success in ideopathic thrombocytopenic purpura had led to a trial of gamma globulin in a number of autoimmune conditions . Success has been reported in myasthenia gravis, rheumatoid arthritis, diabetes, patients with circulating antibodies to factor VIII and Kawasaki's disease . The mechanism of action is unknown but almost certainly multifactorial . Two proven mechanisms that will be added to in the future, include blockade of the Fc receptors on cells of the reticulo-endothelial system and manipulation of immunoregulatory T cells by the presence of anti-idiotypic antibodies in the preparation. Comp Immunol Microbiol Infect Dis, 1987, 10(1), 9 - 23 Partial characterization of a Legionella pneumophila serogroup 1 immunodominant antigenic determinant recognized by a monoclonal antibody . Legionella specific antigenic determinant; Petitjean F et al.; A monoclonal antibody was used to characterize a serogroup 1 specific Legionella pneumophila Philadelphia strain 1 antigenic determinant . A quantitative fluorometric assay was developed to quantitate the antibody sites (2.7 +/- 0.4 X 10(5)) on Legionella bacteria and to determine the physico-chemical parameters of the antibody-antigen interaction (at 4 degrees C: delta G = -10.9 Kcal X mol-1, delta H = 1.7 Kcal X mol-1, delta S = 45 cal X K-1 X mol-1) . The same method was used to study the modification or the removal of the antigen by chemical and enzymatic means (trypsin, papain, lysozyme, acetone, chloroform-methanol and Tris-EDTA); only Tris-EDTA extraction resulted in a significant decrease in antibody binding sites . Inhibition studies of the fluorescein-labelled antibody binding were performed with different sugars of which only L-fucosylamine was inhibitory, and with other monoclonal antibodies to Legionella pneumophila serogroup 1 in order to compare their fine specificity and affinity . The results indicate that the epitope recognized was an immunodominant carbohydrate including an aminodideoxyhexose and carried by the lipopolysaccharide. Annu Rev Pharmacol Toxicol, 1987, 27, 279 - 300 Health effects of exposure to diesel exhaust particles; McClellan RO; Diesel-powered vehicles emit substantially more particles than do gasoline-powered vehicles with contemporary emission control systems . The DEP are submicron in size and readily inhaled . Approximately one-fourth of the particle mass inhaled by people is deposited in the pulmonary region, some of which is retained with a half-life of several hundred days . In animal studies, exposure to high levels of DEP overwhelms the normal clearance mechanisms and results in lung burdens of DEP that exceed those predicted from observations at lower exposure concentrations . A variable amount of the mass of DEP is extractable with strong organic solvents . The extracted material contains more than a thousand individual compounds and is mutagenic in a number of bacterial and mammalian cell assays . Bioassay-directed chemical analysis of DEP had identified several hundred compounds . Many are PAHs, some of which are considered to have human carcinogenic potential . A number of nitrated compounds have been identified that account for a significant portion of the mutagenicity assayed in bacteria . The mutagenicity of the DEPE is generally reduced by addition of an S-9 cellular fraction or of serum proteins . Macrophages rapidly reduce the recoverable mutagenic activity associated with DEP . These findings support a hypothesis that detoxification of DEP-associated organics occurs rapidly in vivo . The association of benzo(a)pyrene and nitropyrene with DEP prolongs their retention in the lungs . This increased retention suggests the need to clarify the relative importance of competing mechanisms that detoxify particle-associated compounds and those that serve to enhance the retention of toxicologically important compounds . Some extracts of DEP evoke tumorigenic responses in skin-tumor bioassays, suggesting their carcinogenic potential in mammals . A number of large-scale studies have been conducted with laboratory rodents to evaluate the effects of chronic inhalation exposure to DE . An increased incidence of lung tumors, some of which were diagnosed as malignant, was observed in 5 studies with rats following exposure for 2 or more years to high levels of DE . Most of the lung tumors were observed after 2 years . Similar studies in Syrian hamsters have yielded negative results . Studies with mice have given mixed results . The results of some studies with laboratory animals exposed to DE and known carcinogens suggest that exposure to DE enhances the effect of the known carcinogens . The specific mechanisms of tumor induction in the DE-exposed rats are unknown . Hypotheses and experimental data have been advanced in support of both genetic and epigenetic mechanisms of action of the DE.(ABSTRACT TRUNCATED AT 400 WORDS) Microbios, 1987, 49(198), 47 - 54 Effects of anti-Ureaplasma urease antibody on homologous and heterologous urease activities; Eng H et al.; Among organisms in the class Mollicutes only Ureaplasma species possess urease . Antiserum to urease of U . urealyticum strain T960 (CX8) was used to examine the cross-reactivity of urease from other Ureaplasma species, as well as urease of jack bean and several urease-possessing walled bacteria . Immunological cross reactivity was used to establish phylogenetic relationships between various antigens . The ability of monospecific anti-urease antibody to inhibit urease activity was examined . The antiserum inhibited urease activity of the homologous strain the least of any Ureaplasma tested . It is postulated that urease possesses a minimum of two sets of epitopes . Binding of antibody to one epitope causes inhibition of enzyme activity; this epitope is common to urease of all Ureaplasma species . Binding of antibody to the other epitope prevents binding to the inhibition epitope; this epitope is specific to U . urealyticum strain T960 (CX8) . No inhibition was observed with urease from jack bean or several walled bacteria. Biochemistry, 1986 Dec 30, 25(26), 8447 - 53 Evidence that heme d1 is a 1,3-porphyrindione; Chang CK et al.; Heme d1 is the noncovalently associated heme prosthetic group of the bacterial nitrite reductase known as cytochrome cd1 . Additional evidence has been obtained in support of a dioxoisobacteriochlorin, or 1,3-porphyrindione, skeleton for this heme . The new data include the natural abundance 13C NMR spectrum of the free base methyl ester derivative of d1, mass spectrometric determinations of the molecular mass of the free base methyl ester and the Cu and the Zn chelates, visible and 1H NMR spectral comparisons between d1 and synthetic porphyrindione model compounds, and the isolation and characterization of several byproducts formed during the purification of the free base methyl ester of d1 . The accumulated evidence strongly supports the following structure for the skeleton of d1: 1-oxo-2-methyl-2'-acetyl-3-oxo-4-methyl-4'-acetyl-5-methyl-6-acrylyl+ ++-7- propionyl-8-methylporphyrin. Cancer, 1986 Dec 15, 58(12), 2640 - 5 Tuberculosis-associated hemophagocytic syndrome . A systemic process; Campo E et al.; Virus-associated hemophagocytic syndrome has been recently identified as a benign, reactive histiocytic proliferation distinct from Rappaport's malignant hystiocytosis . Other etiologic agents, including different bacteria, have also been identified . Three autopsy cases of hemophagocytic syndrome associated to acute tuberculous sepsis are presented . Benign histiocytic proliferation with striking hemophagocytosis was present in a disseminated, multisystemic pattern in all three cases . A relationship between systemic histiocytic proliferation and an anergic status in these patients is suggested. Ann Gastroenterol Hepatol (Paris), 1986 Dec, 22(7), 391 - 3 {Abdominal actinomycosis: a rare complication of gastric surgery? Apropos of a case}; Dellagi K et al.; One case of abdominal actinomycosis is reported which was detected in a diabetic patient three years after surgery for duodenal peptic ulcer . Gastric actinomycosis is very rare and is usually discovered during surgery for gastric mass . The role of surgery as a predisposing condition to infection is discussed . The diagnosis rely on the characterization of the bacteria in cultures from a discharging wall fistula and on histopathologic examination which reveals the characteristic actinomycotic granules. Clin Nephrol, 1986 Dec, 26(6), 303 - 6 Clindamycin phosphate kinetics in subjects undergoing CAPD; Schwartz MT et al.; The kinetics of intraperitoneally administered clindamycin phosphate were studied in 9 volunteer subjects undergoing CAPD . Volunteers were assigned to 2 groups with the first group receiving clindamycin phosphate 300 mg/l in exchanges 1 through 5, and the second group receiving clindamycin phosphate 300 mg/l in exchange 1, and then 30 mg/l in exchanges 2 through 5 . Clindamycin serum and dialysate effluent levels were determined by bioassay . When admixed with dialysate fluid and instilled into the peritoneal cavity, clindamycin phosphate is rapidly activated . Serum concentrations of clindamycin were rapidly achieved in both groups during the first exchange . Subjects in groups I and II had peak serum levels of active drug within 3 (3.94 ug/ml) and 5 (7.35 ug/ml) h, respectively . These results support the practice of not only administering intraperitoneal clindamycin phosphate to treat CAPD-related peritonitis, but using this route of administration to treat systemic infections due to susceptible bacteria in patients without intravenous access. Am J Vet Res, 1986 Dec, 47(12), 2618 - 20 Adherence of Bordetella avium to turkey tracheal mucosa: effects of culture conditions; Arp LH; Adherence of Bordetella avium to the tracheal mucosa of turkeys was evaluated, using bacteria grown under different culture conditions . Several solid and liquid media were used at incubation times of 12, 24, 36, or 48 hours with incubation temperatures of 18, 26.5, or 35 C . Adherence of B avium was greatest when the bacteria were grown on solid media at 35 C . Use of Bordet-Gengou or brain-heart infusion agar was associated with significantly greater (P less than 0.05) adherence compared with adherence of bacteria grown on other media . Adherence was greatest, using cultures in the stationary phase of growth; however, with some media, adherence diminished when incubation was extended beyond 36 hours . Adherence of B avium was reduced but not completely prevented when cultures were incubated at 18 C. Am Rev Respir Dis, 1986 Dec, 134(6), 1287 - 8 Pyrazinamide is not active in vitro against Mycobacterium avium complex; Heifets LB et al.; Strains of M . tuberculosis that elaborate pyrazinamidase are typically susceptible in vitro and in vivo to pyrazinamide (PZA) . However, we found that 33 strains of M . avium complex (MAC), all of which were pyrazinamidase-positive, were resistant in vitro to a high concentration (100 micrograms/ml) of PZA when tested at low pH in 7H12 broth by radiometric (BACTEC) method . The drug was equally ineffective against these bacteria within cultured normal human macrophages . We conclude that the pyrazinamidase test is not suitable for susceptibility studies against M . avium complex . On the basis of our in vitro studies, which support earlier data, we believe that PZA is not appropriate for therapy of MAC disease. Biochem Cell Biol, 1986 Dec, 64(12), 1333 - 8 Sea urchin sperm peroxidase is competitively inhibited by benzohydroxamic acid and phenylhydrazine; Schuel H et al.; Sea urchin sperm contain a phenylhydrazine-sensitive peroxidase that is believed to use hydrogen peroxide produced by the fertilized egg to reduce sperm fertility and thereby assist in the prevention of polyspermy . Strongylocentrotus purpuratus sperm were treated initially with hypotonic phosphate buffer (pH 7.0) to remove catalase and then extracted with 0.5% Triton X-100 in 0.5 M acetate buffer (pH 5.0) . Peroxidase activity in this detergent extract was assayed using 3,3',5,5'-tetramethyl benzidine (TMB) as oxidizable substrate . Kinetic studies showed that the Km for TMB is 250 microM . Benzohydroxamic acid and phenylhydrazine are known to be competitive inhibitors of a variety of plant and animal peroxidases . These substances were found to competitively inhibit the sea urchin sperm peroxidase: for benzohydroxamic acid, Ki = 51.2 microM, mean inhibitory dose (ID50) = 146.7 microM; for phenylhydrazine, Ki = 201 nM, ID50 = 303 nM . These findings indicate that the biochemical properties of the sea urchin sperm peroxidase resembles those of peroxidases found in somatic tissues where oxygen radicals are produced by phagocytes to kill bacteria and support our hypothesis that the sperm peroxidase has a functional role in the prevention of polyspermy during fertilization. Virology, 1986 Dec, 155(2), 593 - 9 Cytoplasmic localization of the HTLV-III 3' orf protein in cultured T cells; Franchini G et al.; HTLV-III, the etiological agent of the acquired immunodeficiency syndrome, contains in its genome coding regions for several novel proteins . One of these, the 3' open reading frame (3'orf) encodes proteins of 26-27 kDa which are expressed in infected cells both in vivo and in vitro . A specific antiserum has been raised against the recombinant 3'orf protein synthesized in bacteria and used to localize the viral proteins by subcellular fractionation and immunofluorescence on HTLV-III infected cells . The antiserum specifically immunoprecipitated the 26- to 27-kDa proteins from both the cytoplasmic (S100) and the membrane fractions, with an enrichment in the latter . The proteins were not detected in the nucleus or organelle (S100 pellet) fractions . These proteins were also recognized in the same subcellular fractions by human sera from patients with AIDS . Indirect immunofluorescence on fixed infected cells confirmed the presence of the proteins in the cytoplasm . Immunoprecipitation and Western blot analysis of total proteins from disrupted HTLV-III virions with the specific antiserum failed to detect the 3'orf protein products, suggesting that they are not a major component of mature virions and may be involved in the intracellular regulation of viral replication. Clin Orthop, 1986 Dec, (213), 211 - 5 Preoperative use of povidone-iodine . A prospective, randomized study; Zdeblick TA et al.; One hundred one adult orthopedic surgical patients were studied in a randomized, prospective clinical trial to compare the effectiveness of the standard povidone-iodine scrub and paint with povidone-iodine painting alone for presurgical skin preparation . No infections occurred in either group . The scrub-plus-paint group showed a 0.601 logarithmic reduction in bacteria counts, compared with 0.622 with painting alone . Further, 36.8% of the patients in the scrub-plus-paint group had skin counts that actually increased after preparation, compared with 13.8% of patients in the paint-only group . The preparation bacterial counts among inpatients, who received preoperative hexachlorophene showers, were significantly lower than that of outpatients, who did not receive preoperative showers . The data support the use of the preoperative hexachlorophene shower and the omission of scrubbing from the surgical skin preparation technique. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1986 Dec, 263(1-2), 133 - 6 A role for interleukin-1 in the pathogenesis of Lyme disease; Beck G et al.; Interleukin-1 (IL-1) is the major immunoregulatory molecule produced by macrophages in response to a variety of environmental insults including chemicals, phagocytosis, bacteria, and bacterial products . Macrophages stimulated by Borrelia burgdorferi produced large quantities of IL-1 when spirochetes were added to macrophages at a ratio of 10 spirochetes per macrophage . The release of IL-1 was dose dependent: a single spirochete per macrophage was sufficient to produce significant quantities of IL-1 . Spirochetal lipopolysaccharide was not required for this activity in that polymyxin B in the spirochete-macrophage culture had no effect on IL-1 production . Normal murine fibroblasts cultured with this IL-1 were shown to have an increased rate of DNA synthesis and an increase in secreted collagenase . IL-1 was found in joint fluids from Lyme disease patients . When IL-1 was injected intradermally into the backs of rabbits, the injection sites became indurated, erythematous, and warm to the touch after 4 hrs and annular lesions much like those of erythema chronicum migrans were seen in some animals after 24 hrs . B . burgdorferi is a powerful inducer for IL-1 in vitro, and it is reasonable to presume that it acts similarly in Lyme disease patients . Our results suggest that IL-1 in turn, may play a role in many of the clinical manifestations of Lyme disease. Zentralbl Bakteriol Mikrobiol Hyg {B}, 1986 Dec, 183(1), 70 - 8 {Recent studies on the recolonization of drinking water}; Schoenen D; About the re- or aftergrowth in drinking water there are some reports . But there are still no real ideas about the reasons and the colony counts to which the water may rise under practical conditions when there is no external influence . In these investigations drinking water was stored in a reservoir for 3-6 weeks and the increase of colonies was observed . In all six tests there was an increase in the colony count . The maximum counts fluctuated between 8-2000 colonies/ml . The water, a mixture of surface- and groundwater, had not a homogeneous tendency of re- or aftergrowth . The chemical parameters gave no indications to these differences. Antimicrob Agents Chemother, 1986 Dec, 30(6), 852 - 5 Evaluation of a radiometric method for pyrazinamide susceptibility testing of Mycobacterium tuberculosis; Tarrand JJ et al.; Pyrazinamide susceptibility testing of Mycobacterium tuberculosis requires an acid environment . By controlling the method of acidification and the quality and quantity of the inoculum, the test can be performed with the BACTEC radiometric system (Johnston Laboratories, Towson, Md.) . We acidified BACTEC 7H12 medium with buffered phosphoric acid and adjusted the test inoculum to 1/10 of that usually employed in BACTEC protocols; after 5 days of growth we correctly identified 36 of 36 strains susceptible to 50 micrograms of pyrazinamide per ml . All 18 resistant strains were classified as pyrazinamide resistant . (Susceptibility or resistance had been determined by standard plate assays.) The test was able to detect small resistant populations in artificial mixtures of 1 or 2% resistant bacteria with a susceptible strain (10 mixtures each) . We tested 70 M . tuberculosis strains in acidified BACTEC 7H12 medium and by the plate dilution test at pH 5.5 . All strains grew in the BACTEC medium, but three strains failed to grow on plates and were not tested further; the results of both methods agreed for the remaining strains. Int J Lepr Other Mycobact Dis, 1986 Dec, 54(4), 578 - 83 Separation of Mycobacterium leprae from contamination with armadillo-liver-derived "pigment" particles; Lloyd GM et al.; Mycobacterium leprae isolated from armadillo liver by the widely used IMMLEP protocol is sometimes contaminated with a particulate "pigment." This paper describes a simple, efficient, and rapid method for purifying large quantities of contaminated bacteria, which may readily be used as an additional step added at the end of the protocol when necessary . The process involves a discontinuous Percoll gradient and generates an essentially pure fraction containing greater than 90% of the original bacteria, and a fraction of "pigment" slightly contaminated with bacteria . Use of the system should release large additional numbers of pure M . leprae suitable for use in human vaccine trials. Antimicrob Agents Chemother, 1986 Dec, 30(6), 883 - 7 Microplate phosphocellulose binding assay for aminoglycoside-modifying enzymes; Cooksey RC et al.; We modified the phosphocellulose binding assay for aminoglycoside-modifying enzymes (AMEs) by use of microdilution plates and a multichannel micropipette . Batteries of aminoglycoside substrates for screening organisms for the presence of AMEs as well as for subclassifying enzymes were prepared and stored in microdilution plates . When tested in parallel with the conventional tube reaction assay, the microplate assay yielded comparable radioactive counts and therefore equally correct identifications of AMEs in 32 isolates representing nine bacterial species . Other modifications, such as multichannel dispensing of crude enzyme preparations and radioisotopic precursors, provided a more rapid, convenient, and less expensive means of examining large collections of organisms for AMEs. Infect Immun, 1986 Dec, 54(3), 811 - 9 Role of mononuclear phagocytes in expression of resistance and susceptibility to Mycobacterium avium infections in mice; Stokes RW et al.; The growth of Mycobacterium avium 702 in the spleens and livers of four inbred strains of mice varied such that the mice could be separated into naturally susceptible (BALB/c and C57BL/6) and naturally resistant (A/Tru and DBA/2) strains . This phenomenon was independent of the size of the infecting inoculum of bacteria in that both low (10(4))- and high (10(7))-dose inocula of M . avium grew progressively in susceptible strains and were eliminated from the target organs of resistant strains . Resistance and susceptibility were also demonstrated in in vitro preparations of macrophages from these strains of mice . Over a 7-day period, replication of M . avium in susceptible mouse macrophages was far greater than that in resistant macrophages . Evidence was obtained to suggest that toxic oxygen metabolites were not responsible for this difference . Though no difference was found in the rate of clearance of M . avium from the blood of susceptible or resistant mice, resident macrophages from susceptible mice ingested more M . avium in vitro than did resident macrophages from resistant animals . Growth of M . avium in spleens of susceptible mice induced a large influx of phagocytes, whereas this was not observed in resistant mice . In contrast to this it was found that, after injection of a variety of inflammatory agents, influx of leukocytes into the peritoneal cavity could not be used to distinguish susceptible and resistant strains of mice. Infect Immun, 1986 Dec, 54(3), 827 - 32 Changes in isoenzyme patterns of a cloned culture of nonpathogenic Entamoeba histolytica during axenization; Mirelman D et al.; The axenization of an Entamoeba histolytica isolate with a nonpathogenic isoenzyme electrophoretic pattern (zymodeme) was recently achieved for the first time (15) . Forty days after the cells were transferred to the medium used for axenic cultivation, the amebae developed virulence properties, and the zymodeme converted to a pathogenic pattern . To exclude the possibility that the original isolate consisted of two zymodeme populations and that conditions of growth selected for a particular population, the experiment was repeated with a cloned culture of a nonpathogenic (zymodeme III) strain, E . histolytica SAW 1734R clAR, isolated by and obtained from P . G . Sargeaunt . Axenization was accomplished, as before, by transferring trophozoites to TYI-S-33 medium containing a mixture of antibiotics to suppress the growth of the associated bacterial flora and a nutritional supplement consisting of gamma-irradiated bacteria . A change in the hexokinase and phosphoglucomutase isoenzyme pattern was observed 21 days after the amebae had been transferred to the axenic medium but before complete axenization of the amebae had occurred . The change in zymodeme was accompanied by an increase in virulence, as evidenced by the ability of fewer amebae to induce hepatic abscesses in hamsters . A reverse conversion to a nonpathogenic zymodeme was also accomplished by reassociating and subculturing the newly converted pathogenic trophozoites of strain SAW 1734R clAR with the bacterial flora that accompanied this ameba in the original xenic culture . The electromobilities of the hexokinase isoenzymes changed back to their original pattern 7 days after the amebae were returned to xenic growth conditions . Our in vitro results demonstrate that culture conditions and bacterial flora can cause changes in the zymodeme and virulence of a cloned ameba isolate and raise the concern that this could happen also in vivo . Thus, the finding of a particular zymodeme in a culture of E . histolytica isolated from a carrier should not be used to predict a clinical condition or serve as a basis for the recommendation of therapy. J Bacteriol, 1986 Dec, 168(3), 1180 - 8 Oxygen-regulated mRNAs for light-harvesting and reaction center complexes and for bacteriochlorophyll and carotenoid biosynthesis in Rhodobacter capsulatus during the shift from anaerobic to aerobic growth; Zhu YS et al.; The stability and regulation by oxygen of mRNAs for the photosynthetic apparatus in Rhodobacter capsulatus have been studied by using proflavin to inhibit transcription and by shifting cells from anaerobic to aerobic conditions . The results from the inhibition experiments show that the mRNA for the light-harvesting LH-II polypeptides (beta, alpha) is more stable than that for the light-harvesting LH-I polypeptides (beta, alpha) during anaerobic growth, whereas the mRNAs for the reaction center polypeptides L (RC-L), M (RC-M), and H (RC-H) are less stable than both the LH-I and LH-II mRNAs . When photosynthetic cells are shifted from anaerobic to aerobic conditions, an immediate decrease in the levels of mRNA for the LH-I, LH-II, RC-L, RC-M, and RC-H proteins was observed . The level of mRNA for the LH-II proteins, however, is more sensitive to oxygen and is reduced faster than the level of mRNA for the LH-I proteins . These results suggest that oxygen represses the expression of genes coding for the light-harvesting antenna and reaction center complexes and may selectively accelerate the degradation of mRNA for the LH-II proteins . The mRNAs for several enzymes in the bacteriochlorophyll biosynthetic pathway are regulated by oxygen in a similar manner . The mRNAs for carotenoid biosynthetic enzymes, however, are regulated by oxygen in a different way . We have found that the amounts of mRNAs for carotenoid biosynthetic enzyme, relative to the amounts of mRNAs for LH and RC, increased during the shift from anaerobic to aerobic conditions . We have particularly shown that although the expression of most photosynthetic genes in R . capsulatus is repressed by oxygen, the crtA gene, located in the BamHI H fragment of the R' plasmid pRPS404 and responsible for the oxidation of spheroidene to spheroidenone, responds to oxygen in an opposite fashion . This exzymatic oxidation may protect the photosynthetic apparatus from photooxidative damage. Postgrad Med, 1986 Nov 15, 80(7), 112 - 9 Hand infections--the academic surgeon's perspective . 2 . A rundown of various causes; Byrne JJ; A large number of organisms can cause infections of the hand, and they can enter the body in a number of ways--in animal or human bites, in punctures, by direct inoculation, or through the bloodstream, intravenous needles, or prosthetic implantation procedures . In addition, all types of scratches and abrasions can become contaminated--by bacteria, viruses, fungi, and a wide variety of marine organisms . When the body's defenses are impaired by disease or other factors, the organisms are more likely to take hold . Treatment depends on the organism involved and the extent of infection, but proper wound care and aseptic technique are paramount in all cases. J Immunol, 1986 Nov 15, 137(10), 3211 - 5 Induction of colony-stimulating activity (CSA) by a synthetic muramyl peptide (MDP): synergism with LPS and activity in C3H/HeJ mice and in endotoxin-tolerized mice; Galelli A et al.; Injection of MDP into mice induces a rapid elevation of monocyte-macrophage CSA in the serum . This effect can also be observed in LPS-hyporesponsive C3H/HeJ mice . MDP and LPS induce CSA synergistically in normal mice . In contrast to the tolerance that is rapidly observed after repeated administration of LPS, MDP does not lose its capacity of inducing serum CSA after repeated injections . Repeated daily injections of MDP also fail to induce tolerance to the LPS-CSA inducing effect . Furthermore, whereas mice rendered tolerant to LPS become hyporesponsive to many other bacteria or bacterial products, they remain responsive to MDP . These data showing that MDP can act synergistically with another CSA inducer, can be injected repeatedly, and can stimulate mice unresponsive to LPS suggest potentially important in vivo applications. Poult Sci, 1986 Nov, 65(11), 2098 - 103 Microelements in the circulation of coccidiosis-infected chicks; Turk DE; Four-week-old Single Comb White Leghorn male chicks were inoculated with either Eimeria acervulina, E . necatrix, E . brunetti, E . tenella, or left uninfected . On Days 1, 3, 5, 6, 7, 8, 9, 10, 14, 21, 28, and 35 after inoculation, blood was removed from five birds in each group; content of copper, zinc, and iron in plasma and total iron-binding capacity were determined . Copper content in plasma was increased during the acute phase of the infections by lower intestinal tract (E . brunetti, E . tenella) infections but not by upper tract (E . acervulina, E . necatrix) infections . Zinc content in plasma was decreased by E . acervulina and E . necatrix infections on the 7th day and was increased by E . acervulina, E . brunetti, and E . tenella infections on the 9th or 10th days . Iron content in plasma was decreased during the acute phase (Days 5 to 7) of E . acervulina, E . brunetti, and E . tenella infections . Total iron-binding capacity was decreased by acute phase E . acervulina and E . brunetti infections . Eimeria tenella infections increased total iron-binding capacity during both the acute and early recovery phases . Plasma content of all components returned to normal by the latter part of the recovery phase of the infections and remained so thereafter . Plasma trace mineral content appeared to be influenced by mineral absorption effects, by hemorrhaging, and perhaps by invasion of the bloodstream by gut bacteria as a result of mucosal damage.(ABSTRACT TRUNCATED AT 250 WORDS) Clin Exp Immunol, 1986 Nov, 66(2), 457 - 62 Development of anti-polysaccharide antibodies in asplenic children; Hammarstrom L et al.; Splenectomized patients are highly susceptible to infections with capsulated bacteria and an impaired response to vaccination with bacterial polysaccharides has frequently been observed in these individuals . Based partly on experimental animal data, an important role for the spleen in the production of specific anti-carbohydrate antibodies, i.e . mostly IgG2 in normal adults, has been suggested . We therefore determined the immunoglobulin class and subclass pattern of serum antibodies from asplenic patients against protein and carbohydrate antigens . Normal levels of total serum IgM, IgG2 and specific IgM and IgG2 anti-polysaccharide antibodies were observed, suggesting only a minor role for the spleen in determining the antibody repertoire . The data imply that the impaired phagocytic capacity and/or the inability to mount a sufficiently rapid antibody response are the main factors underlying the increased susceptibility to bacterial infections noted in these patients. Zh Mikrobiol Epidemiol Immunobiol, 1986 Nov, (11), 94 - 9 {Mechanism of the transmission of the causative agents of infectious diseases}; Kuznetsov VA; Another interpretation, differing from the commonly acknowledged one, of the transfer mechanism is presented . The factors transporting pathogens in the body are the integral part of the transfer mechanism . They ensure the interrelation of the infectious and epidemic processes . In typical contact infections these factors function integrally as the transfer mechanism consisting of two components . This is the sum total of environment and internal factors, specific for a given pathogenic species . These factors ensure the travel of the pathogens from an infected body to a new one . From this viewpoint, the source of infection cannot be an independent element of the infectious process . It is only a transfer factor . The localization of the pathogen, constituting just one of the stages of its travel route in the body, cannot determine the specificity of the transfer mechanism . Its specificity depends on the properties of the pathogen. Genetics, 1986 Nov, 114(3), 717 - 30 General nonchemotactic mutants of Caulobacter crescentus; Ely B et al.; We have examined 35 mutants that have defects in general chemotaxis . Genetic analysis of these mutants resulted in the identification of at least eight che genes located at six different positions on the Caulobacter crescentus chromosome . The cheR, cheB and cheT genes appeared to be located in a three-gene cluster . Mutations in these three genes resulted in the inability of the flagellum to reverse the direction of rotation . Defects in the cheR gene resulted in a loss of the ability to methylate the methyl-accepting chemotaxis proteins . In vitro experiments showed that the lack of in vivo methylation in cheR mutants was due to the absence of methyltransferase activity . Defects in the cheB gene resulted in greatly reduced chemotaxis-associated methylation in vivo and a loss of methylesterase activity in vitro . The specific defects responsible for the lack of a chemotactic response have not been determined for the other identified che genes. J Allergy Clin Immunol, 1986 Nov, 78(5 Pt 1), 887 - 90 Cell-mediated immunity in cat-scratch disease; Gerber MA et al.; The importance of cell-mediated immunity in cat-scratch disease (CSD) is suggested by the positive skin test reactions and granulomatous histopathology noted in patients with this disease . However, an earlier investigation found that lymphocytes from patients with CSD and control subjects were equally unresponsive in vitro to cat-scratch antigen . In contrast, we found that 16 patients with CSD had significantly increased lymphocyte transformation responses to cat-scratch antigen when patients were compared to control subjects . This cell-mediated immune response may be directed against nonviable bacteria in the involved lymph nodes and may be the major mechanism responsible for the granulomatous reaction and clinical features of CSD. J Bacteriol, 1986 Nov, 168(2), 799 - 804 Biochemical and immunochemical properties of the cell surface of Renibacterium salmoninarum; Fiedler F et al.; The biochemical composition of the cell envelope of Renibacterium salmoninarum was investigated in a total of 13 strains isolated from different salmonid fish species at various geographical locations of the United States, Canada, and Europe . A marked similarity with the type strain R . salmoninarum ATCC 33209 was found both in the peptidoglycan and the cell wall polysaccharide . The primary structure of the peptidoglycan was found to be consistent with lysine in the third position of the peptide subunit, a glycyl-alanine interpeptide bridge between lysine and D-alanine of adjacent peptide subunits, and a D-alanine amide substituent at the alpha-carboxyl group of D-glutamic acid in position 2 of the peptide subunit . The cell wall polysaccharide contained galactose as the major sugar component which was accompanied by rhamnose, N-acetylglucosamine, and N-acetylfucosamine . The polysaccharide amounted to more than 60% of the dry weight of the cell walls . It was found to be covalently linked to the peptidoglycan and was released by hot formamide treatment . On gel filtration chromatography the extracted polysaccharide behaved like a homogeneous polymeric compound . The purified cell wall polysaccharide showed antigenic activity with antiserum obtained by immunization of rabbits with heat-inactivated trypsinized cells of R . salmoninarum . Immunoblotting experiments with nontrypsinized cell walls and antisera raised against R . salmoninarum cells revealed that antigenic proteins were attached to the cell walls. Arch Intern Med, 1986 Nov, 146(11), 2173 - 5 Polymicrobial bacterascites . A unique entity in the spectrum of infected ascitic fluid; Runyon BA et al.; A retrospective analysis of 1578 abdominal paracenteses revealed ten cases of polymicrobial bacterascites, ie, growth of multiple organisms in ascitic fluid with a neutrophil count less than 250 cells/cu mm . Six of the ten paracenteses that documented this condition were traumatic (bloody or producing feculent material) . Clinical peritonitis developed in only one patient . No one died as a result of the infection . Polymicrobial bacterascites is rare (0.6% of paracenteses) . It is frequently due to a traumatic paracentesis (bowel entry by the paracentesis needle), and is associated with low morbidity. J Virol, 1986 Nov, 60(2), 525 - 30 Adsorption of mycoplasmavirus MV-L2 to Acholeplasma laidlawii: effects of changes in the acyl-chain composition of membrane lipids; Steinick LE et al.; The enveloped mycoplasmavirus MV-L2 and its host Acholeplasma laidlawii JA1 were used to study the ways in which changes in the membrane lipid bilayer affect virus adsorption . The physical state of the membranes was altered by (i) using viruses and bacteria with different membrane lipid acyl-chain compositions, (ii) using incorporation of cholesterol, and (iii) changing the temperature . Adsorption of viruses was strongly dependent on the acyl-chain composition of the virus and the host . Adsorption to homologous hosts was poor, whereas adsorption to hosts with highly different membrane lipid acyl-chain composition was much stronger . We found a heterogeneity within virus populations produced from hosts with different acyl-chain compositions . In a given virus population, various subpopulations differing in acyl-chain composition were found that differed in their ability to adsorb to cells with a specific acyl-chain composition . The adsorption rate increased slightly when cholesterol was present in the viral membranes but decreased considerably when cholesterol was present in the bacterial membranes . The rate of adsorption was temperature dependent with an increase in adsorption rate above 20 degrees C (for hosts with equal amounts of palmitoyl and oleoyl acyl chains) . MV-L2 did not adsorb to the persistently L2-infected strain JA1(2R) but adsorbed very well to the virus-resistant strain A(EF22) . The physicochemical properties of the lipid matrix of both virus and host are obviously important factors in the adsorption process. Infect Immun, 1986 Nov, 54(2), 347 - 53 Entry of Bartonella bacilliformis into erythrocytes; Benson LA et al.; Bartonella bacilliformis, which causes the human diseases Oroya fever and verruga peruana, binds to human erythrocytes in vitro and produces substantial and long-lasting deformations in erythrocyte membranes, including cone-shaped depressions, trenches, and deep invaginations . The deforming force is probably provided by the polar flagella of these highly motile bacteria . Deep invaginations containing bacteria are commonly seen, and membrane fusion at the necks of the invaginations leads to the formation of intracellular vacuoles containing bacteria . Fluorescent compounds present externally render the vacuoles fluorescent and, occasionally, lightly fluorescent cells are seen, suggesting that the vacuoles sometimes rupture to admit the bacteria to the cytoplasm . Vacuoles present in fluorescent erythrocytes prepared by preloading the erythrocytes with fluorescent compounds are seen as dark areas from which the fluorescent marker is excluded . Entry of the bacteria appears to be the result of a process of forced endocytosis. Exp Cell Res, 1986 Nov, 167(1), 119 - 26 Relationship between cellular diadenosine 5',5'''-P1,P4-tetraphosphate level, cell density, cell growth stimulation and toxic stresses; Segal E et al.; In order to elucidate the postulated role of diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A) in cell growth regulation, the Ap4A cellular content was measured in cells submitted to various treatments affecting the cell growth . Ap4A level was found to increase ten times when cells reached confluence, whereas no significant variation of the ATP pool was observed . Cell growth arrest after serum depletion did not cause any variation in the Ap4A pool . A limited increase in the Ap4A pool was observed when growth of arrested cells was reinitiated but this variation reflected only the increase of cell density . No significant variation in the Ap4A intracellular level was observed after submitting two eukaryotic cell lines to various stresses (cytotoxic drugs, ethanol and heat-shock treatments) . These results suggest that, in eukaryotic cells, Ap4A is not involved in cell growth stimulation but rather is associated with cell contact growth inhibition . They also suggest that Ap4A is not an 'alarmone', contrary to what has been proposed for bacteria. Med Hypotheses, 1986 Nov, 21(3), 249 - 52 The presence of trans unsaturated fatty acids in ruminant meat and milk is unnatural; Booyens J; It is proposed that the presence of large quantities of trans unsaturated fatty acid isomers as final metabolic products of the rumen bacteria and their consequent presence in ruminant tissues and milk is completely unnatural . This results from excessive feeding of domestic ruminants and the consequent passage of bacteria containing partially hydrogenated trans fatty acids into the small intestine . Digestion of such bacteria and the subsequent absorption of the released trans fatty acids could account for their unnatural presence in ruminant tissues and secretion in milk . It is also proposed that the feeding of ruminants, poultry and pigs with unsaturated fatty acid-containing concentrates, could also be a source of trans fatty acids which occur in their tissues. J Clin Periodontol, 1986 Nov, 13(10), 939 - 43 Metronidazole: a double-blind trial in untreated human periodontal disease; Watts T et al.; The effect of metronidazole was assessed in 20 subjects with untreated periodontitis, in a double-blind trial . Subjects received a 1-week course of drug or placebo immediately after baseline examination, and were reassessed 1 month and 3 months later . A 0.25 N constant force probe and darkfield microscopy were used . At 1 month, a difference was observed between the deepest individual pockets in experimental and control groups, with regard to change in probing depth and the proportions of non-motile organisms . At 3 months, differences were observed in 1-3 mm baseline pockets, where a greater proportion of control pockets increased in probing depth, and in 4-6 mm pockets, where a greater proportion of baseline bleeding pockets had ceased to bleed in the experimental group . The differences were of limited clinical significance when compared to studies where metronidazole was used as an adjunct to other therapy, and the results were equivocal with regard to the specific plaque hypothesis, since inhibition of connective tissue attachment loss was not unambiguously demonstrable. J Clin Microbiol . 1986 Nov;24(5):859. Evaluation of a xylene substitute in Ehrlich indole test; Egleton JH et al.; Hemo-De (PMP Medical Industries, Inc., Irving, Tex.) was evaluated and found to be a suitable substitute for xylene in the Ehrlich modification of the indole test. J Dent Res, 1986 Nov, 65(11), 1349 - 52 The effects of the availability of diet on the levels of exoglycosidases in the supragingival plaque of macaque monkeys; Smith K et al.; Supragingival plaque from macaque monkeys was assayed for 13 exoglycosidase enzymes, with appropriate p-nitrophenylglycosides and N-acetylneuramin-lactose used as substrates . Protein in each plaque sample was quantitated with a Coomassie Brilliant Blue dye binding assay, and the specific activity of each enzyme was calculated . In monkeys fed a starch-based diet, fasting resulted in significant increases in the levels of alpha-L-fucosidase, beta-N-acetyl-D-glucosaminidase, beta-N-acetyl-D-galactosaminidase, and neuraminidase . Such changes are consistent with the hypothesis that plaque bacteria degrade salivary glycoproteins for their growth and maintenance in both the presence and especially the absence of dietary food . In contrast, fasting monkeys previously fed a sucrose-rich diet showed no significant alterations in the specific activities of those enzymes whose levels were increased in the starch-based diet group . It is considered likely that, under the conditions prevailing when the sucrose-rich diet is fed, the effective and maximal utilization of sucrose by plaque bacteria necessitates the increased mobilization of nitrogen sources, including the amino sugars of glycoproteins. J Clin Periodontol, 1986 Nov, 13(10)