J Bacteriol, 1992 Mar, 174(5), 1647 - 55 Purification of histidase from Streptomyces griseus and nucleotide sequence of the hutH structural gene; Wu PC et al.; Histidine ammonia-lyase (histidase) was purified to homogeneity from vegetative mycelia of Streptomyces griseus . The enzyme was specific for L-histidine and showed no activity against the substrate analog, D-histidine . Histidinol phosphate was a potent competitive inhibitor . Histidase displayed saturation kinetics with no detectable sigmoidal response . Neither thiol reagents nor a variety of divalent cations had any effect on the activity of the purified enzyme . High concentrations of potassium cyanide inactivated histidase in the absence of its substrate or histidinol phosphate, suggesting that, as in other histidases, dehydroalanine plays an important role in catalysis . The N-terminal amino acid sequence of histidase was used to construct a mixed oligonucleotide probe to identify and clone the histidase structural gene, hutH, from genomic DNA of the wild-type strain of S . griseus . The cloned DNA restored the ability of a histidase structural gene mutant to grow on L-histidine as the sole nitrogen source . The deduced amino acid sequence of hutH shows significant relatedness with histidase from bacteria and a mammal as well as phenylalanine ammonia-lyase from plants and fungi. Gastroenterology, 1992 Mar, 102(3), 1065 - 70 Bacillary peliosis hepatis as a cause of acute anemia in a patient with the acquired immunodeficiency syndrome; Garcia-Tsao G et al.; A 33-year-old white homosexual man, infected with the human immunodeficiency virus, presented with acute anemia and thrombocytopenia not responsive to transfusions or to treatment with steroids and intravenous gamma-globulin . Hematologic workup was compatible with peripheral sequestration or loss of blood cells; however, there was no evidence of gastrointestinal or other sources of hemorrhage, and the only significant finding was a progressive liver enlargement . An abdominal computerized tomographic scan showed a massive homogeneous liver without focal lesions, a very small amount of ascites, and no retroperitoneal fluid collections . A transjugular liver biopsy specimen showed the cystic, blood-filled cavities characteristic of peliosis hepatis . Cavities varied in size, all contained pooled erythrocytes, and some had areas suggestive of thrombi in various stages of organization . Bacteria similar in morphology to those described in bacillary peliosis hepatis were seen in the peliotic spaces . The clinical picture began resolving shortly after treatment with zidovudine and ampicillin/sulbactam was started and had totally resolved 6 months after presentation . This case shows that bacillary peliosis hepatis is a reversible entity that may produce acute sequestration of blood in the liver. Gastroenterology, 1992 Mar, 102(3), 1000 - 8 Bile secretory immunoglobulin A in biliary infection and cholelithiasis; Yio XY et al.; Bile samples from 71 patients with cholelithiasis and a control group of 10 subjects without hepatobiliary diseases were cultured for bacteria and measured for secretory immunoglobulin A (SIgA) using enzyme immunoassay specific for SIgA . The results of bile bacterial culture were all positive in patients with primary bile duct pigment stones, and significantly lower bile SIgA levels were observed than in normal controls (P less than 0.005) . It was also shown that the constitutent ratios of SIgA to total bile immunoglobulin and the bile-serum ratio of SIgA were markedly lower in these patients than in normal controls (P less than 0.001, P less than 0.001) . In patients with cholecystolithiasis, bile SIgA concentrations of patients with biliary infections were remarkably lower than those of patients without biliary infection (P less than 0.01) and those of normal controls (P less than 0.01) . These results suggest a close relationship between biliary tract infection and low concentrations of bile SIgA. J Infect, 1992 Mar, 24(2), 191 - 5 Paecilomyces lilacinus infection in a child with chronic granulomatous disease; Silliman CC et al.; Chronic granulomatous disease (CGD) is a rare inherited disorder in which neutrophils do not appropriately generate cytotoxic superoxide anion, the respiratory burst, in response to invading bacteria or fungi as a part of normal host defence . We report the case of a child with CGD who had two abdominal wall abscesses caused by Paecilomyces lilacinus, an organism not previously known to cause infections in patients with CGD . The abdominal wall is a location that is rarely associated with Paecilomyces infections . Parenteral amphotericin B eradicated the infection in an immunocompromised child whereas this regimen has heretofore largely been unsuccessful in the treatment of this infection . Paecilomyces species and other fungi from immunocompromised hosts and thought to be laboratory contaminants, need to be carefully investigated for they may become pathogens in this clinical setting. J Neuroimmunol, 1992 Mar, 37(1-2), 149 - 60 Serotonergic aspects of the response of human platelets to immune-adjuvant muramyl dipeptide; Polanski M et al.; The human platelet serotonergic responses of aggregation and uptake were shown to be modulated by muramyl peptides . Muramyl dipeptide inhibited serotonin uptake in a temperature dependent and stereospecific manner . It also blocked the binding to platelets of {3H}imipramine, a well-known inhibitor of serotonin uptake . Muramyl dipeptide decreased serotonin2 (5-HT2) mediated platelet aggregation, and blocked the binding of a 5-HT2-specific ligand: lysergic acid diethylamide . Since muramyl peptides are released upon degradation of bacteria, the findings offer a possible mechanism for neuro-immune modulation by emphasizing the interaction between 5-HT (a neurotransmitter) and muramyl peptides (immuno-adjuvants). In Vivo, 1992 Mar-Apr, 6(2), 205 - 9 Inhibition of biological effects of endotoxins by phenothiazines; Molnar J et al.; The importance of change transfer reaction involving endotoxins of bacteria and interactions between endotoxin-induced tumor necrosis factor (TNF) were investigated both in vitro and in vivo in the presence of several phenothiazines . Complex formation between endotoxins and ring-substituted phenothiazines, benzodiazepines, amantadine and promethazine was measured using spectrophotometric methods . The endotoxin-induced hypotensive effect was prevented by phenothiazine pretreatment of the dogs; however, the endotoxin-phenothiazine complex had the same effect as the endotoxin itself . The tumor necrosis factor-inducing ability of endotoxin in human monocytes was prevented by promethazine . The TNF induction by endotoxin was inhibited by promethazine in vivo. Biophys J, 1992 Mar, 61(3), 694 - 703 Dynamics of energy transfer and trapping in the light-harvesting antenna of Rhodopseudomonas viridis; Zhang FG et al.; By low intensity picosecond absorption spectroscopy it is shown that the exciton lifetime in the light-harvesting antenna of Rhodopseudomonas (Rps.) viridis membranes with photochemically active reaction centers at room temperature is 60 +/- 10 ps . This lifetime reflects the overall trapping rate of the excitation energy by the reaction center . With photochemically inactive reaction centers, in the presence of P+, the exciton lifetime increases to 150 +/- 15 ps . Prereducing the secondary electron acceptor QA does not prevent primary charge separation, but slows it down from 60 to 90 +/- 10 ps . Picosecond kinetics measured at 77 K with inactive reaction centers indicates that the light-harvesting antenna is spectrally homogeneous . Picosecond absorption anisotropy measurements show that energy transfer between identical Bchlb molecules occurs on the subpicosecond time scale . Using these experimental results as input to a random-walk model, results in strict requirements for the antenna-RC coupling . The model analysis prescribes fast trapping (approximately 1 ps) and an approximately 0.5 escape probability from the reaction center, which requires a more tightly coupled RC and antenna, as compared with the Bchla-containing bacteria Rhodospirillum (R.) rubrum and Rhodobacter (Rb.) sphaeroides. Dev Comp Immunol, 1992 Mar-Jun, 16(2-3), 95 - 101 Formation of brown bodies in the coelomic cavity of the earthworm Eisenia fetida andrei and attendant changes in shape and adhesive capacity of constitutive cells; Valembois P et al.; The formation of brown bodies in the coelomic cavity may result from an aggregation of coelomocytes around offending foreign cells such as bacteria, gregarines, incompatible graft fragments, and altered self structures such as setae or necrotic muscle cells . The initial nodule rapidly increases in volume by aggregation of new coelomocytes and various waste particles . When a brown body has reached a diameter of 1-2 mm, its external cells flatten and lose their adhesiveness toward free coelomocytes or waste particles and its pigment rapidly darkens . Brown bodies play an important role in homeostasis. Plant Cell, 1992 Mar, 4(3), 273 - 82 Mutations in the D1 subunit of photosystem II distinguish between quinone and herbicide binding sites; Ohad N et al.; The structure-activity relationships of the plastoquinone QB binding domain in the D1 subunit of photosystem II (PSII) were investigated by characterization of mutations introduced in the D1 protein . Eight novel point mutations in the gene psbA, which encodes D1, were generated in the cyanobacterium Synechocystis PCC6803 by site-specific mutagenesis in vitro . The effects of the resulting modifications in D1 on electron transfer in PSII and on herbicide binding were analyzed . The results extend the structural analogies between the secondary quinone binding site in D1 and in subunit L of the photosynthetic reaction center in purple bacteria . The involvement of Phe255, Ser264, and Leu271 of D1 in plastoquinone binding and electron transfer in PSII was established . An indirect effect of Tyr254 on the binding of QB was demonstrated . Changes in binding of herbicides and QB to D1 as a result of the mutations revealed specific interactions between amino acid residues in D1 and the plastoquinone and distinguished between the binding sites of QB and herbicides. Khirurgiia (Mosk), 1992 Mar, (3), 81 - 4 {The combined treatment of acute suppurative diseases of the fingers and hand using decamethoxin}; Fishchenko AIa et al.; The authors discuss the results of complex treatment of 286 patients with acute pyoinflammatory diseases of the fingers and hand with the use of a new Soviet-produced antiseptic decametoxin . Panaris was diagnosed in 196 (68.5%), phlegmons and abscesses in 82 (29.7%), furuncle in 6 (2.1%) and carbuncle in 2 (0.7%) patients . 224 (78.4%) patients received out-patient and 62 (21.6%) in-patient treatment . The authors established that as the result of the applied complex treatment with the use of various antiseptic compositions containing decametoxin the mean duration of treatment was 7.8 days . The article discusses the causes of the disease, the methods of operative treatment, and management of patients in the postoperative period. J Med Assoc Thai, 1992 Mar, 75 Suppl 2, 20 - 3 Pre-operative shaving and wound infection in appendectomy; Rojanapirom S et al.; The study on the effects of shaving the skin on wound infections after appendectomy was done in 80 patients in 1988 in Siriraj Hospital . Patients were divided randomly into two groups, 40 patients each, a control and an experiment group . The control group had their skin shaved and the experiment group did not . Data showed no difference in: demography, preoperative admission time, interval between skin preparation and surgery, thickness of subcutaneous tissue, operating time, and suture materials . Shaving of the skin resulted in no alteration in bacteria found on the skin, on the walls of the wound before closing . Stitch abscesses were the only wound infection found in 3 patients in each group . It is concluded that skin shaving, though it did not increase wound infection rate, had no beneficial effect on wound infection in appendectomy. Dev Comp Immunol, 1992 Mar-Jun, 16(2-3), 85 - 93 Biological characteristics of the Calliphora vomitoria agglutinin; McKenzie AN et al.; The galactose specific agglutinin from Calliphora vomitoria was found to be expressed in the haemolymph of all the larval instars, but could not be detected at any other time during the life cycle . The haemagglutinating activity was insensitive to wounding of the tegument or injection of saline; however, a significant increase in haemagglutinating titre could be induced upon inoculation of the haemocoel with biotic or abiotic particulate material . The agglutinin also actively agglutinated several bacterial species and appeared capable of playing a role in particle--haemocyte interaction . The presence of the purified agglutinin significantly increased the attachment of fetuin-derivatized beads to haemocytes in vitro, and this activity could be specifically reduced by the addition of galactose, suggesting that the agglutinin may act as an opsonin. Mol Cell Biol, 1992 Mar, 12(3), 991 - 7 SH2 domains of the p85 alpha subunit of phosphatidylinositol 3-kinase regulate binding to growth factor receptors; McGlade CJ et al.; The binding of cytoplasmic signaling proteins such as phospholipase C-gamma 1 and Ras GTPase-activating protein to autophosphorylated growth factor receptors is directed by their noncatalytic Src homology region 2 (SH2) domains . The p85 alpha regulatory subunit of phosphatidylinositol (PI) 3-kinase, which associates with several receptor protein-tyrosine kinases, also contains two SH2 domains . Both p85 alpha SH2 domains, when expressed individually as fusion proteins in bacteria, bound stably to the activated beta receptor for platelet-derived growth factor (PDGF) . Complex formation required PDGF stimulation and was dependent on receptor tyrosine kinase activity . The bacterial p85 alpha SH2 domains recognized activated beta PDGF receptor which had been immobilized on a filter, indicating that SH2 domains contact autophosphorylated receptors directly . Several receptor tyrosine kinases within the PDGF receptor subfamily, including the colony-stimulating factor 1 receptor and the Steel factor receptor (Kit), also associate with PI 3-kinase in vivo . Bacterially expressed SH2 domains derived from the p85 alpha subunit of PI 3-kinase bound in vitro to the activated colony-stimulating factor 1 receptor and to Kit . We infer that the SH2 domains of p85 alpha bind to high-affinity sites on these receptors, whose creation is dependent on receptor autophosphorylation . The SH2 domains of p85 are therefore primarily responsible for the binding of PI 3-kinase to activated growth factor receptors. Proc Natl Acad Sci U S A, 1992 Mar 1, 89(5), 1539 - 43 Intrinsic anion channel activity of the recombinant first nucleotide binding fold domain of the cystic fibrosis transmembrane regulator protein; Arispe N et al.; The first nucleotide binding fold (NBF-1) from the cystic fibrosis transmembrane regulator (CFTR) has been expressed in bacteria and found to bind ATP and to express anion channel activity when reconstituted onto a planar lipid bilayer . This evidence suggests that the NBF forms the anion-selective portion of the CFTR channel . We also found that the recombinant NBF-1 anion channel is blocked by ATP (1 mM), under which condition it appears to have a minimal conductance of approximately 9 pS and an ohmic current-voltage relationship . We further found that the recombinant NBF-1 bearing the delta F508 mutation has nearly identical anion channel activity to that of the wild-type protein but can be distinguished from wild type under bianionic conditions with chloride and gluconate . We conclude from these data that the anion channel activity of the recombinant NBF-1 could represent all or part of the anion conductance mechanism of CFTR and that the role of the ATP binding by the NBF could be to modulate this anion channel activity. Arch Oral Biol, 1992 Mar, 37(3), 167 - 73 Dipeptidyl peptidase II- and IV-like activities in gingival tissue and crevicular fluid from human periodontitis lesions; Cox SW et al.; Gingival tissue and gingival crevicular fluid were collected from patients with chronic periodontitis . Gel filtration chromatography of crude tissue extracts yielded separate fractions active against Lys-Ala-7-amino-4-trifluoromethyl-coumarylamide (AFC) at acid pH and Gly-Pro-AFC at alkaline pH . The molecular weights, pH optima and inhibitor responses of these activities were consistent with those of dipeptidyl peptidases (DPP) II and IV, respectively . When tested with the same substrates, crevicular fluid was also found to contain DPP II- and IV-like activities with very similar pH profiles and inhibitor responses to those in tissue . The close resemblance suggested that the crevicular fluid enzymes were derived mainly from inflamed gingival tissues . Slight differences in the DPP II-like activities might be explained by the additional presence in crevicular fluid of enzymes from subgingival bacteria . With use of appropriate buffers, a third substrate, Ala-Pro-AFC, gave selective detection of both DPP II- and IV-like activities in tissue and crevicular fluid . Assays with Ala-Pro-AFC had the advantage of greater sensitivity, especially with DPP II-like activity . Raised levels of this enzyme have previously been found in the gingiva of periodontitis patients and thus DPP II-like activity in crevicular fluid might prove of value in monitoring disease activity. J Hosp Infect, 1992 Mar, 20(3), 193 - 8 Laboratory evaluation of a filter for the control of cross-infection during pulmonary function testing; Kirk YL et al.; Nosocomial transmission of respiratory pathogens is a possible complication of lung function testing . The use of a filter, placed between the patient and the spirometer equipment may be one way of preventing such nosocomial spread . This paper reports the laboratory evaluation of the efficiency of such a filter at removing bacteria from exhaled breath . Volunteers exhaled 100 normal breaths or four forced exhalations through a filter on to a blood agar plate . Bacterial counts on the agar plate were compared with recoverable bacterial counts from the filter . Total challenge from the forced exhalations ranged from 161-84,200 colony forming units . The calculated efficiency of 10 volunteers was 99.9% . For the normal breaths the challenge ranged from 0-262,000 colony forming units . There was no growth on any of the blood-agar plates . These filters appear to be highly efficient at removing exhaled bacteria. Mol Phylogenet Evol, 1992 Mar, 1(1), 26 - 30 Phylogenetic relationships of the endosymbionts of mealybugs (Homoptera: Pseudococcidae) based on 16S rDNA sequences; Munson MA et al.; A portion of the gene coding for the 16S ribosomal RNA from the endosymbionts of three species of mealybugs {Pseudococcus longispinus (Targioni-Tozzetti), Pseudococcus maritimus (Ehrhorn), and Dysmicoccus neobrevipes (Beardsley)} was cloned, sequenced, and compared to a homologous fragment from bacteria representative of aphid endosymbionts as well as major subdivisions of the Proteobacteria . Parsimony analysis of the sequences indicated that the mealybug endosymbionts are related and belong to the beta-subdivision; in contrast, previous studies showed that aphid endosymbionts are part of the gamma-subdivision . These findings suggest that the endosymbiosis of mealybugs is a consequence of a single bacterial infection and indicate that this ancestor was different from the ancestor involved in aphid endosymbiosis. Rev Med Chil, 1992 Mar, 120(3), 304 - 10 {Fournier's disease}; Bannura G; Fournier's disease or perineal gangrene is a severe necrotizing process affecting the ano-rectal, perineal and scrotal region which is usually secondary to ano-rectal pathology and periurethral or cutaneous processes . Ample surgical debridement, wide spectrum systemic antibiotics and general supportive measures are essential for survival in this very serious disease. Res Microbiol, 1992 Mar-Apr, 143(3), 341 - 5 Hybridization of transposon Tn501 for detection of mercury resistance sequences in a marine environment; Prabu SK et al.; Total genomic DNA isolated by concentrating seawater and mercury-resistant bacteria were hybridized with a mer probe to detect the presence of homologous DNA sequences in marine coastal waters of the Bay of Bengal, India . Coastal water extracts induced with mercury hybridized with the mer operon of transposon Tn501 . Most of the mercury-resistant bacteria that volatilized mercury also contained homologous DNA sequences to the mer probe. J Dent Res, 1992 Mar, 71(3), 453 - 7 Silica-induced precipitation of calcium phosphate in the presence of inhibitors of hydroxyapatite formation; Damen JJ et al.; The promotion and the inhibition of hydroxyapatite formation by various substances were determined by measurement of the induction time of spontaneous precipitation (ti) from supersaturated solutions . Silica was found to decrease ti in Hepes-buffered (pH 7.2) supersaturated solutions with a wide range of calcium-to-phosphate ratios and concentrations . Also, in suspensions of the oral bacteria S . mutans or C . matruchotii in 1 mmol/L calcium, 7.5 mmol/L phosphate, and 50 mmol/L Hepes (pH 7.2), silica was capable of stimulating precipitation . Macromolecules derived from these bacteria by freezing and thawing appeared to be strong inhibitors of calcium phosphate precipitation . In the presence of silica, the effects of these bacterial inhibitors could be partially overcome, which supports the idea that silica in dental plaque is a promoter of calculus formation . In contrast, inhibition of calcium phosphate precipitation by a low-molecular-weight inhibitor, pyrophosphate, could not be counteracted by silica. Medicine (Baltimore), 1992 Mar, 71(2), 59 - 72 Anti-Hu--associated paraneoplastic encephalomyelitis/sensory neuronopathy . A clinical study of 71 patients; Dalmau J et al.; We studied 71 patients with "paraneoplastic" encephalomyelitis, sensory neuronopathy, or both associated with the presence of the anti-Hu antibody in their serum . Most (78%) had small-cell lung cancer . In 9 patients no tumor was detected . Fifty-two patients (73%) had signs and symptoms of multifocal involvement of the nervous system; in 28 (39%), 2 areas, and in 24 (34%), 3 or more areas were clinically affected . Sensory neuronopathy was present in 52 patients (74%), but in only 44 (62%) did it dominate the course of the disease . Other predominant findings were: motor neuron dysfunction (14 patients, 20%), limbic encephalopathy (14, 20%), cerebellar symptoms (11, 15%), brainstem encephalopathy (10, 14%), and autonomic nervous system dysfunction (7, 10%) . The presence of the anti-Hu antibody prompted a search for the tumor in 60% of the patients; the tumor when found was usually small and remained localized until death, or was demonstrated only at autopsy . Treatment using steroids and plasmapheresis, immunosuppressants, or both, did not improve the paraneoplastic symptoms . Autonomic and respiratory failure, either of central origin or secondary to neuromuscular weakness, were the principal causes of death . Patients with rapidly developing sensory neuropathy or symptoms of encephalomyelitis should be studied for the presence of the anti-Hu antibody; if the antibody is found, the possibility of small-cell lung cancer should be investigated . If a tumor is not found in the initial search, one may become evident in several months. J Leukoc Biol, 1992 Mar, 51(3), 264 - 9 Fc receptor expression, concanavalin A capping, and enzyme content of bovine neonatal neutrophils: a comparative study with adult cattle; Zwahlen RD et al.; The increased susceptibility of newborns to infection may in part be related to impaired in vitro functions of neonatal polymorphonuclear neutrophils (PMNs) . To evaluate early steps in the activation cycle of bovine PMNs we determined the expression of Fc receptors (FcRs) with an erythrocyte rosetting assay utilizing bovine anti-sheep immunoglobulin G2 IgG2 and the accumulation of ligand receptor complexes or "caps" with fluorochrome-coupled concanavalin A (Con A caps) on neutrophils from adult (A-PMN) and newborn (N-PMN) bovines . In addition, the levels of myeloperoxidase (MPO) and alkaline phosphatase (AP) were determined . FcR expression is reduced in N-PMNs (P less than .001), in contrast to results observed with human N-PMNs . Basal capping of Con A binding sites is reduced (P less than .05) in N-PMNs but is enhanced (P less than .001) upon pretreatment with colchicine (0.5, 5.0, and 50.0 microns) . These findings are again contrary to results observed with human N-PMNs . Consistent with findings in human neonates, however, are reduced levels of cellular MPO (P less than .05) and elevated cellular AP (P less than .001) in the neonate . The functional significance of elevated AP levels and altered Con A capping in N-PMNs is unclear . However, diminished expression of FcR could potentially contribute to impaired adherence and phagocytosis of bacteria, and reduced activity of neutrophil MPO could indicate weaker microbicidal capacity of neonatal cells . The demonstrated impairment of N-PMN functions could potentially contribute to reducing the effectiveness of the cellular host defense system in neonatal calves. Biochemistry, 1992 Feb 18, 31(6), 1886 - 91 Inhibition of protein kinase C by annexin V; Schlaepfer DD et al.; Annexin V is a protein of unknown biological function that undergoes Ca(2+)-dependent binding to phospholipids located on the cytosolic face of the plasma membrane . Preliminary results presented herein suggest that a biological function of annexin V is the inhibition of protein kinase C (PKC) . In vitro assays showed that annexin V was a specific high-affinity inhibitor of PKC-mediated phosphorylation of annexin I and myosin light chain kinase substrates, with half-maximal inhibition occurring at approximately 0.4 microM . Annexin V did not inhibit epidermal growth factor receptor/kinase phosphorylation of annexin I or cAMP-dependent protein kinase phosphorylation of the Kemptide peptide substrate . Since annexin V purified from both human placenta and recombinant bacteria inhibited protein kinase C activity, it is not likely that the inhibitor activity was associated with a minor contaminant of the preparations . The following results indicated that the mechanism of inhibition did not involve annexin V sequestration of phospholipid that was required for protein kinase C activation: similar inhibition curves were observed as phospholipid concentration was varied from 0 to 800 micrograms/mL; the extent of inhibition was not significantly affected by the order of addition of phospholipid, substrate, or PKC, and the core domain of annexin I was not a high-affinity inhibitor of PKC even though it had similar Ca2+ and phospholipid binding properties as annexin V . These data indirectly indicate that inhibition occurred by direct interaction between annexin V and PKC . Since the concentration of annexin V in many cell types exceeds the amounts required to achieve PKC inhibition in vitro, it is possible that annexin V inhibits PKC in a biologically significant manner in intact cells. Gene, 1992 Feb 15, 111(2), 157 - 63 Dictyostelium discoideum as an expression host for the circumsporozoite protein of Plasmodium falciparum; Fasel N et al.; We have used the cellular slime mold, Dictyostelium discoideum (Dd), to express the Plasmodium falciparum circumsporozoite protein (CS), a potential component of a subunit vaccine against malaria . This was accomplished via an expression vector based on the discoidin I-encoding gene promoter, in which we linked a sequence coding for a Dd leader peptide to the almost complete CS coding region (pEDII-CS) . CS production at both the mRNA and protein levels is induced by starving cells in a simple phosphate buffer . Variation in pH or cell density does not seem to influence CS synthesis . CS-producing cells can be grown either on their normal substrate, bacteria, or on a semi-synthetic media, without affecting CS accumulation level . The CS produced in Dd seems similar to the natural parasite protein as judged by its size and epitope recognition by a panel of monoclonal antibodies . We constructed a second expression vector in which the CS is under the control of a Dd ras promoter . CS accumulation can then be induced by external addition of cAMP . Such a tightly regulated promoter may allow expression of proteins potentially toxic to the cell . Thus, Dd could be a useful eukaryotic system to produce recombinant proteins, in particular from human or animal parasites like P . falciparum. Blood, 1992 Feb 15, 79(4), 888 - 94 Rapid recovery from cytopenia in hairy cell leukemia after treatment with 2-chloro-2'-deoxyadenosine (CdA): relation to opportunistic infections; Juliusson G et al.; Sixteen patients with symptomatic hairy cell leukemia were treated with a single course of 2-chloro-2'-deoxyadenosine (CdA), 0.7 mg/kg total dose . Twelve patients achieved complete remission (CR) . One patient with a CD19+/CD5+/CD25- phenotype and one with a pentostatin-treated CD19+/CD25- variant form had minor responses . Two patients with advanced disease and poor performance status died early from invasive mycosis . Three patients recovered from infections caused by cytomegalovirus and by candida . No patient had infections caused by bacteria or by unknown organisms . The median time to full recovery from anemia and thrombocytopenia was 6 and 2 weeks from start of therapy, respectively . Patients with infections, however, recovered at 13 and 5 weeks, respectively . Neutrophil, monocyte, and lymphocyte counts returned to normal at a median of 5, 5, and 10 weeks, respectively . Infections developed more frequently in pancytopenic patients than in those with one or more blood cell count within the normal range (P less than .01) . All patients with one or no previous therapy had a CR, whereas those with more than one previous regimen had a lower CR rate (P less than .01) . Thus, 1 week of CdA therapy frequently induced CR also in patients resistant to interferon . Toxicity was limited, and recovery from cytopenia was faster than what is reported during interferon therapy. J Immunol Methods, 1992 Feb 14, 147(1), 73 - 81 Species differentiation of mycoplasmas by EF-Tu specific monoclonal antibodies; Kamla V et al.; Ten mouse hybridoma cell lines producing IgG monoclonal antibodies to mycoplasmal elongation factor Tu (EF-Tu) were established . These mAbs showed different degrees of cross-reactivity between mollicutes and even other bacteria . This finding, indicating protein structure diversities of pan bacterial EF-Tu should permit species differentiation of mycoplasmas by epitope pattern analysis of a single protein . Epitope patterns of 23 mollicute type strains and of 40 M . hominis isolates were determined by ELISA . All M . hominis patterns were found to be closely related whereas intrageneric patterns differed in a species specific manner. Am J Trop Med Hyg, 1992 Feb, 46(2), 105 - 15 Cell-mediated immune responses of adults to vaccination, challenge with Rickettsia rickettsii, or both; Dumler JS et al.; As a part of a study to evaluate a formalin-killed Rickettsia rickettsii vaccine, lymphoproliferative (LT) and delayed-type hypersensitivity (DTH) skin test responses to killed R . rickettsii were measured as correlates of cell-mediated immunity in volunteers who were vaccinated, challenged with R . rickettsii, or both . We detected LT responses in 26 (51%) of 51 volunteers after vaccination . After challenge, six of six unvaccinated volunteers and 12 of 16 vaccinated volunteers developed Rocky Mountain spotted fever (RMSF); all 22 mounted LT responses . The vaccinated individuals developed LT responses of greater magnitude and 1-2 weeks earlier than unimmunized controls (41,049 versus 15,084 mean net counts per minute {cpm}), suggesting that vaccination primed the cellular immune system . Moreover, development of LT responses postvaccination was associated with the amelioration of RMSF, as indicated by a slightly longer mean incubation period (328 hr versus 302 hr) and a shorter illness (19 hr versus 26 hr) in LT responders than in LT nonresponders . However, the postvaccination LT response did not discriminate between vaccinated individuals who resisted challenge and those who did not . Skin tests using killed R . rickettsii as antigen, performed in volunteers 14-17 months postvaccination or 12-15 months after challenge, revealed a weak but significant reaction in 50% of those who had received vaccine only, and a moderately strong reaction in all vaccinated and unvaccinated volunteers who had been challenged with R . rickettsii . The relationships between induction of protective immunity against intracellular bacteria by killed and replicating organisms and LT and DTH responses are discussed. Nucleic Acids Res, 1992 Feb 11, 20(3), 501 - 6 Directional recombination is initiated at a double strand break in human nuclear extracts; Lopez BS et al.; The involvement of a double strand break in the initiation of homologous recombination was examined in human nuclear extracts . M13 duplex derivatives, containing inserts in the LacZ' region (producing white plaques), were cleaved by restriction enzymes and coincubated in the extracts with a circular plasmid containing the LacZ' region without insert, and unable to produce plaques . Repair was estimated by the ability to produce plaques after transfection into JM109 (recA1) bacteria . Recombination with the plasmid enhances the number of plaques and also the frequency of M13 producing blue plaques . Heterologous insertions in the region surrounding the break were analyzed for their effects on initiation of recombination . The extent of repair by recombination (number of plaques) was compared with the number of blue plaques among the repaired population . Initiation of recombination is inhibited when heterologous insertions are located at 7bp from the break, on the right side as well as on the left side . A low level of recombination is measurable for 27 bp of homology but the maximum efficiency of recombination occurred with homologies of 165 or 320 bp from the break to the heterologous insertion . At 320 bp, the extent of recombinational repair remained at a plateau level but the frequency of blue plaques progressively decreases . We have also analyzed the effect of different sizes of inserts . With longer inserts, a longer length of homology adjacent to the break is required for optimum recombination . However, the size of the insert does not affect the low level of recombination that occurred with a short homology (27 bp) . The results indicate that the process is initiated at or near the break, requires homology on both sides of the break and is followed by an elongation from the double strand break to the distal regions of the DNA . Our data provide some support to the double-strand-break repair model established for meiotic recombination in yeast. J Trauma, 1992 Feb, 32(2), 179 - 82 Necrotizing fasciitis of the extremities; Wang KC et al.; Necrotizing fasciitis is a limb- and life-threatening soft-tissue infection . Eighteen patients with necrotizing fasciitis of the extremities were reviewed . These infections occurred most commonly after minor trauma . Associated chronic debilitating diseases were present in 13 patients . All but two infections were polymicrobial . The overall mortality rate was 33% . Death was caused by persistent wound sepsis in three and systemic septic complications in spite of apparent local infection control in three . At the admission physical examination the condition may resemble a benign, low-grade cellulitis . Three of four patients died because of a delay in surgical debridement for more than 24 hours after admission . Multiple radial debridements of the involved skin, fat, and fascia are essential to control progressive necrosis . Our results suggest that early diagnosis and prompt surgical debridement of necrotizing fasciitis are essential for survival. Proc Natl Acad Sci U S A, 1992 Feb 1, 89(3), 839 - 42 Materials derived from biomass/biodegradable materials; Luzier WD; Interest in biodegradable plastics made from renewable resources has increased significantly in recent years . PHBV (polyhydroxybutyrate-polyhydroxyvalerate) copolymers are good examples of this type of materials . This paper provides an overview of the manufacturing process, properties, biodegradability, and application/commercial issues associated with PHBV copolymers . They are naturally produced by bacteria from agricultural raw materials, and they can be processed to make a variety of useful products, where their biodegradability and naturalness are quite beneficial . PHBV copolymers are still in the first stage of commercialization . But they are presented in this paper as an example of how new technology can help meet society's needs for plastics and a clean environment. Proc Natl Acad Sci U S A, 1992 Feb 1, 89(3), 1021 - 5 Effects of different DNA polymerases in ligation-mediated PCR: enhanced genomic sequencing and in vivo footprinting; Garrity PA et al.; We have developed a simplified procedure for the ligation-mediated polymerase chain reaction (LMPCR) using Thermococcus litoralis DNA polymerase (Vent DNA polymerase) . We show that Vent DNA polymerase produces correct, blunt-ended primer extension products with substantially higher efficiency than Thermus aquaticus (Taq) DNA polymerase or modified T7 DNA polymerase (Sequenase) . This difference leads to significantly improved genomic sequencing, methylation analysis, and in vivo footprinting with LMPCR . These improvements include representation of all bands with more uniform intensity, clear visualization of previously difficult regions of sequence, and reduction in the occurrence of spurious bands . It also simplifies the use of DNase I cut DNA for LMPCR footprinting. Infect Immun, 1992 Feb, 60(2), 653 - 6 Topographic mapping of Helicobacter pylori colonization in long-term-infected pigs; Engstrand L et al.; Four barrier-born pigs were inoculated with Helicobacter pylori during gastroscopy . Infection in all pigs was established after 3 weeks, and the animals were kept isolated from other pigs in ordinary experimental sites . The pigs were sacrificed and examined 3, 5, 6, and 6.5 months postinoculation . A detailed urease mapping of the pig stomachs showed a patchy distribution of H . pylori . The bacteria colonized in all pigs, with a concentration of H . pylori-positive areas in the antrum and fundus . Furthermore, the number of colonized areas tended to increase with time, and some of these areas showed a strong urease reaction, indicating a heavy colonization with H . pylori . Biopsies from these areas contained 10(2) to 10(5) CFU per 2-mm-wide biopsy . We conclude that persistence of H . pylori infection in barrier-born pigs can be demonstrated for at least 6.5 months . The patchy distribution and the variability of viable bacteria were similar to those described for humans. Appl Environ Microbiol, 1992 Feb, 58(2), 724 - 6 Comparison of assimilable organic carbon and UV-oxidizable carbon for evaluation of ultrapure-water systems; Governal RA et al.; Bacterial growth potential was measured in an ultrapure-water pilot plant by modified assimilable organic carbon (AOC) and UV-oxidizable carbon tests . An ion-exchange unit increased UV-oxidizable carbon, yet did not significantly (P greater than or equal to 0.05) alter AOC values . UV radiation decreased UV-oxidizable carbon and increased AOC. Appl Environ Microbiol, 1992 Feb, 58(2), 584 - 92 Measurement of marine picoplankton cell size by using a cooled, charge-coupled device camera with image-analyzed fluorescence microscopy; Viles CL et al.; Accurate measurement of the biomass and size distribution of picoplankton cells (0.2 to 2.0 microns) is paramount in characterizing their contribution to the oceanic food web and global biogeochemical cycling . Image-analyzed fluorescence microscopy, usually based on video camera technology, allows detailed measurements of individual cells to be taken . The application of an imaging system employing a cooled, slow-scan charge-coupled device (CCD) camera to automated counting and sizing of individual picoplankton cells from natural marine samples is described . A slow-scan CCD-based camera was compared to a video camera and was superior for detecting and sizing very small, dim particles such as fluorochrome-stained bacteria . Several edge detection methods for accurately measuring picoplankton cells were evaluated . Standard fluorescent microspheres and a Sargasso Sea surface water picoplankton population were used in the evaluation . Global thresholding was inappropriate for these samples . Methods used previously in image analysis of nanoplankton cells (2 to 20 microns) also did not work well with the smaller picoplankton cells . A method combining an edge detector and an adaptive edge strength operator worked best for rapidly generating accurate cell sizes . A complete sample analysis of more than 1,000 cells averages about 50 min and yields size, shape, and fluorescence data for each cell . With this system, the entire size range of picoplankton can be counted and measured. Chem Pharm Bull (Tokyo), 1992 Feb, 40(2), 479 - 83 Homeostasis as regulated by activated macrophage . I . Lipopolysaccharide (LPS) from wheat flour: isolation, purification and some biological activities; Nishizawa T et al.; Based on our new concept of ontogenic inflammation, we have sought a substance which can prime macrophage in terms of the endogenous production of tumor necrosis factor (TNF) . A lipopolysaccharide (LPSw) was found in wheat flour, purified and characterized . The molecular size of LPSw was about 5 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it contained 3-deoxy-D-manno-octulosonic acid: 1, hexosamine: 4 and one phosphorus in a single molecule . LPSw can prime macrophage to release TNF when given intradermally, percutaneously or even orally in mice as well as in humans, in exactly the same way as intravenous administration of interferon gamma. J Clin Periodontol, 1992 Feb, 19(2), 113 - 7 The immediate- and medium-term effects of Meswak on the composition of mixed saliva; Gazi MI et al.; Chewing sticks or Meswaks are used for teeth cleaning in many parts of the world . They contain substances that may reduce caries and periodontal disease . The present study consisted of 2 parts . In a short-term experiment, volunteers chewed on an inert eliciting agent (pyrogen-free rubber) and then a piece of Meswak, each for 5 min . For the medium-term experiment, volunteers brushed with either Meswak or a conventional toothbrush 5 x a day for 2 weeks . Saliva produced immediately after chewing Meswak showed statistically significant increases in calcium and chloride, but decreases in phosphate and pH as compared with controls . In the medium-term experiment, saliva samples collected 4 h after the last use of Meswak or toothbrush showed no significant differences in any of the components examined (calcium, magnesium, chloride, phosphate, IgA, IgG, lactate dehydrogenase and aspartate transaminase) . Gingival and plaque indices, however, were significantly lower after brushing with Meswak . Salivary calcium promotes mineralization of tooth enamel and chloride inhibits calculus formation . Our results thus indicate that Meswak releases substances into saliva that could improve oral health . Calcium and chloride values were similar to those of controls after 4 h and thus frequent use of Meswak may be necessary to maintain a favorable salivary environment. J Clin Periodontol, 1992 Feb, 19(2), 103 - 12 Metronidazole in periodontitis: reduced need for surgery; Loesche WJ et al.; A considerable amount of circumstantial evidence indicates that most forms of periodontitis are due to the presence or dominance of a finite number of bacterial species in the subgingival plaque . Almost all of the putative pathogens are anaerobic species, indicating that most forms of periodontitis could be diagnosed as anaerobic infections . In this double-blind investigation, patients with elevated proportions or levels of spirochetes in 2 or more plaque samples, i.e., 60% spirochetes, were randomly assigned to receive either metronidazole, 250 mg 3 x a day for 1 week, or placebo (positive-control) after the completion of all debridement procedures . When the patients were re-examined 4 to 6 weeks later, the patients in the metronidazole group (n = 15) exhibited a highly significant (p less than 0.01) reduction in probing depth and apparent gain in attachment levels relative to the patients (n = 18) in the positive-control group about those teeth that initially had probing depths of 4 to 6 mm . This pattern was also observed about teeth that initially had probing depths greater than or equal to 7 mm . This reduction in probing depths and apparent gain in attachment was associated with a significant reduction in the need for periodontal surgery in the metronidazole-treated patients (difference 8.4 teeth per patient) compared to the positive-control patients (2.6 teeth per patient) . These clinical improvements in the metronidazole group were associated with significantly lower proportions of spirochetes, selenomonads, motile rods, and P . intermedius, and a significantly higher proportion of cocci in the plaques . These findings indicate that systemic metronidazole, when given after all the root surface debridement is completed, leads to additional treatment benefits, including a reduced need for surgery, beyond that which can be achieved by debridement alone. Aust N Z J Surg, 1992 Feb, 62(2), 161 - 3 Ovarian actinomycosis presenting as acute peritonitis; Dawson JM et al.; A patient with acute peritonitis due to ovarian actinomycosis and the association of this disease with the intrauterine contraceptive device is discussed . Because adequate treatment requires prolonged antibiotics the diagnosis should be sure . Unfortunately if antibiotics are given pre-operatively culture is usually unsuccessful and thus careful examination of all removed material is essential . As many of these women are of child bearing age surgical intervention should aim to preserve fertility wherever possiblePIP: Upon admission to Box Hill Hospital in Victoria, Australia, a 38-year old woman was pale and febrile (328.6 degrees Celsius) and had a pulse of 88 beats/minute . She had had midabdominal pain for 1 week and severe lower abdominal pain for 2 days . Her menses were heavy . Other than pain during examination, rectal and vaginal examinations were normal . She had considerable neutrophilia (leukocyte count = 21.2 x 1 billion) . The X-ray revealed free fluid . Ultrasonography indicated an IUD which she had had for 10 years, a mass with small cystic areas near the right ovary, and fluid in the rectouterine pouch . The physicians suspected peritonitis and administered iv broad spectrum antibiotics (1 mg ampicillin, 80 mg gentamicin, and 500 mg metronidazole) every 8 hours . They did a laparotomy . An abscess containing much green pus, the necrotic right ovary, and the appendix, which appeared normal and later shown not to be infected, occupied the right iliac fossa . The tubes were fine . The surgeons removed the appendix and right ovary . They washed out the abdomen with saline and inserted a drain to the right iliac fossa . The woman improved immediately so the physicians stopped antibiotics 3 days after surgery . Histological tests revealed actinomycosis caused by fast-growing aerobic bacteria which is known to cause necrosis, fibrosis, and suppuration . During recovery, the physicians removed the IUD and performed dilation and curettage . Actinomyces normally just dwell in the mouth and intestines, but, in this case, probably migrated up the IUD tail after spreading from the bowel to the perineum to the vagina . The physicians suspected that the presence of Mycoplasma hominis provided the mucosal breach needed to permit actinomyces' invasion . Physicians should consider actinomycosis in acute abdominal sepsis cases with a longterm use of an IUD . They can treat it with antibiotics since Actinomyces tend to be sensitive to broad spectrum antibiotics . Circ Shock, 1992 Feb, 36(2), 134 - 9 Alterations in intestinal permeability and blood flow in a new model of mesenteric ischemia; Horton JW; Disturbances in intestinal circulation for even short periods of time can produce mucosal injury, translocation of gut bacteria, and multiple organ failure . We recently reported a model of intestinal ischemia that included occlusion of the superior mesenteric artery (SMA) and interruption of collateral arcades from the right colic and jejunal arteries for 20 min . This present study was designed to characterize further our model of intestinal ischemia by quantitatively assessing changes in intestinal permeability (plasma to luminal clearance of 51Cr-labeled EDTA) and intestinal blood flow (IBF) (microspheres) . A total of 89 rats were included for study; mean arterial blood pressure and acid-base balance were not significantly altered by intestinal ischemia or reperfusion . Baseline measurements of 51Cr-labeled EDTA were not significantly different among the experimental animals, and clearance did not change throughout the experimental period in the sham-ischemic group (N = 14) . Clearance of 51Cr-labeled EDTA at the end of 20 min of intestinal ischemia (0.194 +/- 0.057 ml/min/100 gm, N = 17) was significantly greater than that measured at control (0.079 +/- 0.006 ml/min/100 gm, P less than 0.05) . In addition, clearance measurements during reperfusion (20 min, 0.362 +/- 0.051; 60 min, 0.267 +/- 0.084 ml/min/gm) were significantly higher than those measured at the end of ischemia . Baseline IBF was similar in all rats (N = 42); SMA occlusion reduced IBF by 99% from baseline (from 1.4 +/- 0.27 to 0.014 +/- 0.001 ml/min/gm, N = 20) . Removal of the SMA clip returned intestinal perfusion to baseline values (1.72 +/- 0.51 ml/min/g).(ABSTRACT TRUNCATED AT 250 WORDS) Zentralbl Bakteriol, 1992 Feb, 276(3), 398 - 406 Delayed type hypersensitivity (DTH): quantitative studies of mycobacteria-induced destructive and non-destructive DTH-reactions in the skin of guinea pigs; Abou-Rebyeh H et al.; Research on delayed type hypersensitivity (DTH) to facultative intracellular bacteria in guinea pigs focused especially on intracutaneous DTH reactions . However, phenomena of erythematous and necrotic skin reactions were only evaluated qualitatively, dose-response relationships not investigated, differences between early and late reactions overlooked and the development of unspecific skin necrosis ignored . In this work, these aspects were investigated by characterisation of DTH-mediated reactions against mycobacterial antigens in the skin of guinea pigs . Intracutaneous injection of mycobacterial antigen into the skin of immunised and non-immunised guinea pigs induced an erythematous reaction which reached its peak after 24 h . On days two and three after challenge, the centre of the erythema blanched and necrotised . During the next days, skin necrosis increased and reached a peak on days six and seven . In immunised guinea pigs, DTH led to a specific enhancement of erythematous and necrotising skin reactions which also appeared in non-immunised guinea pigs . Thus, an early erythematous and a late necrotising DTH-reaction were identified by their morphology and time course . Both types of DTH-mediated allergy revealed dose-dependency from the intracutaneously injected antigen doses . Erythematous and necrotising DTH reactions against viable and killed mycobacteria showed a similar time course and morphology . DTH reactivity which was induced by immunisation with Mycobacterium tuberculosis showed cross-reactivity with antigens of Mycobacterium bovis. Acta Odontol Scand, 1992 Feb, 50(1), 57 - 64 Oral health behavior of 6-year-old Danish children; Petersen PE; In Denmark the Public Dental Health Service (PDHS) is now in the process of evaluation and revision of oral health education programs . The purpose of the present survey was 1) to evaluate the pattern of oral health behavior among 6-year-old children in relation to family and social characteristics; 2) to describe the level of dental knowledge and attitudes among the parents; and 3) to estimate the relative effect on caries experience of social and behavioral risk factors . The study comprised 212 children (response, 73%), and the parents responded to self-administered questionnaires . Moreover, information on def-s and DMF-S was collected from the epidemiologic recording systems for the PDHS . With regard to dental caries, 98% of the parents were aware of the harmful effect of sugar, and 88% knew about the role of bacteria . The causal effect of bacteria in relation to periodontal disease was stressed by 81% . Most of the parents (93%) believed that the dental diseases are preventable by means of proper oral hygiene habits, restriction of sugar and sweets, and the use of fluorides . The high level of dental knowledge among the parents was related to information given by the PDHS . Toothbrushing at least twice a day was performed by 88% of the children, and most used fluoridated toothpaste . Practical support to the children was given by 45% of the parents, and 55% checked the teeth . On a daily basis, the children had healthy foods like vegetables (59%), fruits (87%), and milk (89%), and orangeade was consumed by 53%; 45% of the children consumed sweets on a specific weekday ('Saturday sweets').(ABSTRACT TRUNCATED AT 250 WORDS) Int Dent J, 1992 Feb, 42(1), 3 - 11 Effect of acid etching on the dental pulp in adhesive composite restorations; Fujitani M et al.; Pulpal response to marginal enamel etching or both enamel and dentine etching with 37% phosphoric acid for adhesive composite restorations was studied in the teeth of the Japanese monkey (Macaca fuscata) . Dentine etching irritated the pulp and caused moderate to severe initial changes along the odontoblastic layer at 3 days . When no bacteria were present along the cavity walls, the subsequent pulpal reactions at 30 and 90 days decreased with time and large amounts of irritation dentine were formed . These inflammatory reactions were less than those caused by enamel etching only, or zinc oxide/eugenol cement . Significant correlation (P less than 0.01) was found between the intensity of the inflammatory reactions and the degree of bacterial infection . The pulpal irritation caused by acid etching of dentine for composite restorations is transitory only when the fillings have a good marginal seal and wall adaptation. Rheum Dis Clin North Am, 1992 Feb, 18(1), 11 - 21 HLA-B27 and disease: a consequence of inadvertent antigen presentation? Benjamin R, Parham P. The close association of HLA-B27 with arthritic conditions has led to the suggestion that these diseases are mediated by cytotoxic T lymphocytes that recognize self-peptides presented by HLA-B27 molecules . The further association with enteric bacterial infections suggests that bacterial antigens may prime the CTL that later crossreacts on self . Bacterial infections do not usually generate CTL responses . We speculate here that unusual properties of HLA-B27 molecules may predispose to such responses . Thus, HLA-B27-related disease may be an unfortunate consequence of the generation of a suitable, self-mimicking HLA-B27-binding peptide by certain bacteria, plus an unusual propensity for the HLA-B27 molecule to bind and present such peptides. J Dairy Sci, 1992 Feb, 75(2), 399 - 405 Bovine neutrophil responses to parenteral vitamin E; Hogan JS et al.; Twenty-eight Holsteins were tested to determine effects of dietary and parenteral vitamin E supplementation during the dry period on plasma alpha-tocopherol and in vitro neutrophil functions at calving . Cows were assigned to one of four experimental groups receiving either supplemental dietary vitamin E, injections of vitamin E, both dietary and injections of vitamin E, or neither source of supplemental vitamin E during the dry period in a 2 x 2 factorial arrangement . Cows receiving parenteral vitamin E were injected subcutaneously with 3000 IU of vitamin E (dl-alpha-tocopherol) at 10 and 5 d prior to anticipated calving . Cows not receiving parenteral vitamin E were injected with a placebo . Experimental groups receiving dietary vitamin E during the dry period were supplemented with 1040 IU/d compared with none for controls . Cows injected with vitamin E had greater plasma alpha-tocopherol concentration 5 d after the first injection, at calving, and 1 wk after calving than did cows injected with placebo . Plasma alpha-tocopherol concentrations did not differ between dietary vitamin E treatment groups from calving through 4 wk postpartum . No interaction was found between dietary and parenteral supplementation of vitamin E on plasma alpha-tocopherol concentration . Neutrophils from cows injected with vitamin E had greater intracellular kill of bacteria at calving than did neutrophils from placebo-injected cows . Neither phagocytic index nor percentage of neutrophils phagocytizing differed between vitamin E-injected and placebo-injected cows . Dietary vitamin E during the dry period had no effect on neutrophil function at calving . Intracellular kill and plasma alpha-tocopherol were correlated at calving. J Dairy Res, 1992 Feb, 59(1), 11 - 9 Trials of a novel mastitis sensor on experimentally infected cows; Lake JR et al.; Numerous attempts to develop in-line systems for the measurement of the electrical conductivity of milk as an aid to the detection of mastitis have been described . The major technical problem of fouling of electrodes exposed to milk has been overcome in the sensor tested here by use of an inductive system of measurement . The practical application of this system in the milking parlour is reported . The novel mastitis sensor has been shown to detect incipient mastitis before there are visible signs of abnormal milk . Conductivity measurements have the advantage over other diagnostic procedures in that the information is available immediately without additional effort . Normalized conductivity and milk yield variation results indicate the potential for developing a practical, automatic system for mastitis detection. J Mol Evol, 1992 Feb, 34(2), 95 - 114 Directional mutation pressure, selective constraints, and genetic equilibria; Sueoka N; Rates of substitution mutations in two directions, v {from an A-T or T-A nucleotide pair (AT-pair) to a G-C or C-G nucleotide pair (GC-pair)} and u {from a GC-pair to an AT-pair}, are usually not the same . The net effect, v/(u + v), has previously been defined as directional mutation pressure (mu D), which explains the wide interspecific variation and narrow intragenomic heterogeneity of DNA G + C content in bacteria . In this article, first, a theory of the evolution of DNA G + C content is presented that is based on the equilibrium among three components: directional mutation pressure, DNA G + C content, and selective constraints . According to this theory, consideration of both u and v as well as selective constraints is essential to explain the molecular evolution of the DNA base composition and sequence . Second, the theory of directional mutation pressure is applied to the analysis of the wide intragenomic heterogeneity of DNA G + C content in multicellular eukaryotes . The theory explains the extensive intragenomic heterogeneity of G + C content of higher eukaryotes primarily as the result of the intragenomic differences of directional mutation pressure and selective constraints rather than the result of positive selections for functional advantages of the DNA G + C content itself. J Antibiot (Tokyo), 1992 Feb, 45(2), 147 - 50 Nannochelins A, B and C, new iron-chelating compounds from Nannocystis exedens (myxobacteria) . Production, isolation, physico-chemical and biological properties; Kunze B et al.; Novel citrate-hydroxamate siderophores, named nannochelins A, B and C, were isolated from the culture broth of the myxobacterium Nannocystis exedens strain Na e485 . The new substances showed weak growth-inhibitory activity against some bacteria and fungi. J Periodontol, 1992 Feb, 63(2), 87 - 92 Serial doxycycline and metronidazole in prevention of recurrent periodontitis in high-risk patients; Aitken S et al.; The efficacy of metronidazole and doxycycline in preventing recurrent periodontitis was studied in 23 patients . After treatment in the previous 7 months with either bimonthly scaling and 3 weeks of systemic doxycycline (11 subjects) or scaling and placebo (12 subjects), patients were monitored for recurrent periodontitis and were scaled every 2 months . When either a periodontal abscess or greater than 2 mm loss of gingival attachment was observed, metronidazole was administered (250 mg every 8 hours) for 10 days . In the placebo plus metronidazole group, 5 patients (42%) exhibited recurrent periodontitis after the metronidazole regimen compared with only one (9%) in the doxycycline plus metronidazole group (P less than 0.096) . Subgingival plaque samples at study and healthy control sites were screened for the presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Eikenella corrodens, and Fusobacterium nucleatum by immunofluorescence and for spirochetes using Ryu's stain . Presence/absence analysis of the sum of scores of the 6 individual pathogens demonstrated large reductions (P less than 0.005) in the frequency of detection of pathogens in the former doxycycline compared with the placebo plus metronidazole group at both study and control sites before and one month after metronidazole . By 7 months after metronidazole, there was no detectable difference between groups . These results indicate that prevention of recurrent periodontitis with metronidazole may be enhanced by previous treatment with doxycycline. J Dent, 1992 Feb, 20(1), 3 - 10 Microleakage; Taylor MJ et al.; A critical review of techniques used in the assessment of microleakage in dental restorations is presented . These techniques include the use of air pressure, bacteria, radioisotopes, electrochemistry, chemical tracers and dye penetration . Wide variations in methodologies are revealed. J Anim Sci, 1992 Feb, 70(2), 465 - 9 Lean color characteristics of bullock and steer beef; Shackelford SD et al.; Twelve young bulls and 12 steers were slaughtered and the carcasses were fabricated at 48 h postmortem . Top rounds, bottom rounds, and rib eyes were vacuum-packaged and stored at 0 degrees C until 21 d postmortem . After subprimal storage, steaks were removed from each cut and displayed under simulated retail conditions for 3 d . Consumer panelists (n = 110) showed little difference in buying preference for top round, bottom round, and rib eye steaks from steers vs bullocks . Also, consumer panelists could not detect color differences between steaks from steers and bullocks . Experienced panelists (n = 6) detected brighter (P less than .05) muscle color for top round and rib eye steaks from steers but detected no difference in muscle color of bottom round steaks from bullocks and steers . Generally, overall desirability scores favored steaks from steer carcasses . This was particularly true of rib eye steaks . Sex-class of carcass did not affect aerobic plate counts of top round, bottom round, or rib eye steaks either before or after 3 d of retail display . Muscle pH was lower for top round and rib eye steaks from steer carcasses; however, the magnitude of this difference was not large enough to affect retail display stability . Muscle pH of bottom round steaks was not affected by sex-class . Although experienced panelists could detect differences in visual appearance of beef retail cuts from steers and bullocks, consumer panelists did not show a clear purchase preference for steer vs bullock beef . These data suggest that beef retail cuts from bullocks are acceptable in visual appearance. Aliment Pharmacol Ther, 1992 Feb, 6(1), 67 - 77 A comparison of indomethacin with ibuprofen on gastrointestinal mucosal integrity in conventional and germ-free rats; Melarange R et al.; The effects of indomethacin and ibuprofen on gastrointestinal mucosal integrity were studied in conventional and germ-free rats . Only ibuprofen induced significant gastric erosion formation in both conventional and germ-free animals, demonstrating that the presence of micro-organisms is not required in this form of damage . Both indomethacin and ibuprofen caused significant intestinal damage and blood loss in germ-free animals . However, in the conventional counterparts, damage due to indomethacin was enhanced whereas that induced by ibuprofen was not . The results from the present work would suggest that non-steroidal anti-inflammatory drugs such as indomethacin, which are secreted largely in the bile, unlike ibuprofen, may act in concert with bacteria and the constituents of bile to induce, in part, intestinal damage and blood loss. Arch Surg, 1992 Feb, 127(2), 163 - 7; discussion 167-9 Selective gut decontamination reduces nosocomial infections and length of stay but not mortality or organ failure in surgical intensive care unit patients; Cerra FB et al.; Suppression of the gut luminal aerobic flora to reduce nosocomial infections was tested in a prospective, randomized, double-blind, placebo-controlled clinical trial in patients in a surgical intensive care unit who had persistent hypermetabolism . Forty-six patients were randomized to receive either norfloxacin, 500-mg suspension every 8 hours, together with nystatin, 1 million units every 6 hours, or matching placebo solutions administered through a nasogastric tube within 48 hours of surgical intensive care unit admission . Selective gut decontamination with the experimental therapy or placebo solutions continued for at least 5 days or until the time of surgical intensive care unit discharge . Patients were monitored with routine surveillance cultures for the development of nosocomial infections, as defined by criteria from the Centers for Disease Control . All other therapy was given as clinically indicated, including systemic antibiotics . The selective gut decontamination group experienced a significant reduction in the incidence of nosocomial infections and a reduced length of stay . However, these results were not associated with a concomitant decrease in progressive multiple organ failure syndrome, adult respiratory distress syndrome, or mortality. Mol Gen Genet, 1992 Feb, 231(3), 494 - 8 The nifU, nifS and nifV gene products are required for activity of all three nitrogenases of Azotobacter vinelandii; Kennedy C et al.; Strains with mutations in 23 of the 30 genes and open reading frames in the major nif gene cluster of A . vinelandii were tested for ability to grow on N-free medium with molybdenum (Nif phenotype), with vanadium (Vnf phenotype), or with neither metal present (Anf phenotype) . As reported previously, nifE, nifN, nifU, nifS and nifV mutants were Nif- (failed to grow on molybdenum) while nifM mutants were Nif-, Vnf- and Anf- . nifV, nifS, and nifU mutants were found to be unable to grow on medium with or without vanadium, i.e . were Vnf- Anf- . Therefore neither vnf nor anf analogoues of nifU, nifS, nifV or nifM are expected to be present in A . vinelandii. J Clin Microbiol, 1992 Feb, 30(2), 479 - 84 Clinical and epidemiological importance of typing of Mycobacterium avium complex isolates; Tsang AY et al.; The results of the application of a range of typing procedures to the identification and classification of 6,264 cultures of nontuberculous mycobacteria from human sources and the environment are reported . Seroagglutination, an enzyme-linked immunosorbent assay applied to whole bacteria or the glycolipid typing antigens and based on serovar-specific polyclonal or monoclonal antibodies, thin-layer chromatography of these antigens, and gas chromatography of their specific sugar determinants were used to arrive at identifications . As a result of this comprehensive approach, 4,452 (71%) of all cultures and 88% of those of samples from patients with AIDS proved to be typeable . The rank order of frequency of occurrence of individual organisms within the entire group of isolates was Mycobacterium avium complex serovar 4 greater than serovar 8 greater than serovar 1 greater than serovar 9 greater than serovar 6 greater than serovar 14 greater than serovar 2 greater than M . fortuitum greater than M . kansasii greater than M . xenopi greater than an apparent mixture of serovar 4 and M . xenopi greater than a mixture of serovar 4 and serovar 8 . These results were similar but not identical to the pattern observed for isolates obtained from patients with AIDS; the order was M . avium complex serovar 4 greater than serovar 8 greater than serovar 1 greater than a mixture of serovar 4 and M . xenopi, a mixture of serovar 4 and serovar 8 greater than serovar 9 greater than serovar 2 greater than serovar 6 . Serotyping was also used to demonstrate the possible clinical significance of nontuberculous mycobacteria recovered from different body sites . Other information on the distribution of M . avium serovars in patients from different geographical environments is provided. J Clin Microbiol, 1992 Feb, 30(2), 465 - 72 Development of a polymerase chain reaction-based diagnosis of Trichomonas vaginalis; Riley DE et al.; We developed a polymerase chain reaction (PCR)-based test for detecting the protozoan parasite Trichomonas vaginalis . Genomic libraries were constructed from two independent clinical isolates of T . vaginalis . From these libraries, 12 genomic clones were purified, sequenced, and then screened for uniqueness by computer-assisted sequence comparisons . PCR reactions were performed to evaluate eight PCR-primer pairs, including a primer pair that targeted the T . vaginalis ferredoxin gene . All eight primer pairs yielded PCR products of the expected sizes . However, six of the primer pairs amplified their respective target sequences in limited numbers of clinical T . vaginalis isolates, suggesting the presence of significant genomic variability among isolates . An exception was a primer pair, termed TVA5-TVA6, that amplified a 102-bp genomic sequence, termed A6p, in all of 24 clinical isolates . The A6p sequence was not detected by PCR in human DNA or in a wide variety of flagellates, ciliates, or bacteria tested . The A6p sequence appears highly selective for a broad range of T . vaginalis isolates and holds promise for PCR-based diagnosis of the parasite. J Vet Med Sci, 1992 Feb, 54(1), 37 - 42 Protective effect by cell-free antigen obtained from culture supernatant of phase I Bordetella bronchiseptica; Ohgitani T et al.; The cell-free antigen (CFA), with highly hemagglutination activity, obtained from the culture supernatant of Bordetella bronchiseptica was compounded with oil adjuvant to make a component vaccine (CFAV) . In the immunization trial in mice, the offsprings whose mothers were immunized with CFAV escaped from death when challenged intrapleurally with virulent strain of B . bronchiseptica . The protective indices (difference of LD50 dose of the challenge strain between immunized and control groups) of the offsprings from CFAV-immunized mothers were over 3.0 in common logarithm value . Moreover, about 90% of the offsprings from CFAV-immunized mothers were negative in nasal turbinate atrophy, while over 80% of them from non-immunized mothers showed obvious turbinate atrophy when challenged intranasally with virulent strain . On the one hand, remarkable differences in the number of bacteria recovered from nostrils were observed between both test groups . It was concluded that CFAV is a very effective vaccine against B . bronchiseptica infection in animals. Oral Microbiol Immunol, 1992 Feb, 7(1), 24 - 7 Electron immunocytochemical localization of Actinobacillus actinomycetemcomitans leukotoxin; Berthold P et al.; The Actinobacillus actinomycetemcomitans leukotoxin was localized in A . actinomycetemcomitans bacteria using an electron immunocytochemical thin-section labeling method . An immuno-dot blot procedure was initially used to ascertain the optimal specimen fixation . This consisted of a periodate-lysine-paraformaldehyde (2%) fixative in a phosphate buffer followed by embedding in LR White . Affinity-purified toxin was used to produce a monospecific polyclonal antibody . The reaction sites were visualized with a colloidal gold-tagged reporter antibody . The leukotoxin was found to be localized either in the cell envelope and/or in membranous vesicles on the outer surface of the bacterial cell . These results support previous observations indicating the same location. Lik Sprava, 1992 Feb, (2), 57 - 60 {The sterility of blood products manufactured in Ukraine}; Liubich VV; The author reports results of examination for sterility of 3100 series (7343 samples) of hemocorrectors that were received for State control from zonal blood transfusion stations in the Ukraine from 1987 through 1990 . Four of them proved non-sterile . Strict surveillance for aseptics and antiseptics at all stages of the technological process will ensure sterility of all hemotransfusion media obtained from the blood transfusion services. Kansenshogaku Zasshi, 1992 Feb, 66(2), 189 - 93 {An isolation procedure of Chlamydia pneumoniae and Chlamydia trachomatis}; Kazuyama Y; The author devised a method which permits simultaneous isolation and identification of Chlamydia using a slide chamber with 8 wells . Contaminating bacteria were eliminated by filtration with the membrane filter . The procedure allowed isolation of Chlamydia pneumoniae and Chlamydia trachomatis from 11 (10.1%) out of 109 and 10 (9.2%) out of 109 otolaryngologic clinical specimens, respectively . The use of HL cells together with Hela-229 cells had been considered essential to isolate Chlamydia from specimens in this field . The conditions for isolating Chlamydia pneumoniae were similar to those for Chlamydia trachomatis . The slide chamber procedure with membrane filter treated specimens was suitable for simultaneous isolation and culture of various species of Chlamydia and allowed easy differentiation of Chlamydia pneumoniae from Chlamydia trachomatis with identification by the indirect fluorescent-antibody technique . Routine examination of clinical specimens using this procedure should be required in the future in order to better understand chlamydial infections. Zhonghua Wai Ke Za Zhi, 1992 Feb, 30(2), 88 - 90, 124-5 {Biliogenic liver abscess caused by acute obstructive suppurative cholangitis}; Zhong DC; Biliogenic liver abscess was found by autopsy in 52 of 61 (85.2%) cases died of acute obstructive suppurative cholangitis (AOSC) at our hospital from 1957 to 1980 . Of the 52 cases with liver abscess, 44 (84.6%) had multiple abscesses and 47 (90.4%) suffered complications of the rupture of liver abscess . Liver abscess was clinically diagnosed in only 7 of 52 cases who underwent emergency operation, with the predeath definite diagnosis of 13.5% (7/52) . The authors considered biliogenic liver abscess an inevitable outcome not an accidental complication of AOSC when the high pressure in bile duct could not be relieved, and emphasized the importance of prevention, early diagnosis and treatment of liver abscess in order to decrease the mortality of AOSC. Kekkaku, 1992 Feb, 67(2), 113 - 7 {Application of FDA/EB staining for the detection of viable or non-viable mycobacteria in clinical specimens}; Harada S et al.; Two-hundred sputum specimens from tuberculosis patients were examined for viable or non-viable mycobacteria by a combination of fluorescein diacetate ethidium bromide (FDA/EB) staining, Ziehl-Neelsen staining, and the use of cultures in 3% Ogawa egg medium . The sputum specimens were treated with 3% NaOH for 10 min and washed in PBS . The bacteria was then harvested by centrifugation at 6,000 rpm for 5 min . Each sample was subjected to FDA/EB staining, Ziehl-Neelsen staining and cultures to compare the staining -method results and the results of colony formation . Ziehl-Neelsen staining method revealed acid-fast bacteria in the specimens, distributed from Gaffky 1 to Gaffky 8 . The number of FDA-positive specimens and culture-positive specimens were identical in all Gaffky grades, suggesting that the FDA staining method well reflected the presence of viable mycobacteria in the specimens . We concluded that FDA staining is a valuable method to detect viable mycobacteria in sputum specimens on the first day of examination . It is therefore advantageous for doctors and patients to be immediately informed of culture results rather than waiting for several weeks. Gene, 1992 Feb 1, 111(1), 119 - 24 Analysis of a ribosomal RNA operon in the actinomycete Frankia; Normand P et al.; The organisation of ribosomal RNA-encoding (rrn) genes has been studied in Frankia sp . strain ORS020606 . The two rrn clusters present in Frankia strain ORS020606 were isolated from genomic banks in phage lambda EMBL3 by hybridization with oligodeoxyribonucleotide probes . The 5'-3' gene order is the usual one for bacteria: 16S-23S-5S . The two clusters are not distinguishable by restriction enzyme mapping inside the coding section, but vary considerably outside it . Sequencing showed that the 16S-rRNA-encoding gene of ORS020606 is very closely related to that of another Alnus-infective Frankia strain (Ag45/Mut15) and highly homologous to corresponding genes of Streptomyces spp . Two possible promoter sequences were detected upstream from the 16S gene, while no tRNA-encoding gene was detected in the whole operon . Regions with a high proportion of divergence for the study of phylogenetic relationships within the genus were looked for and found in the first intergenic spacer, in the 23S and in the 16S gene. J Clin Microbiol, 1992 Feb, 30(2), 265 - 74 Characterization of a novel Rochalimaea species, R . henselae sp . nov., isolated from blood of a febrile, human immunodeficiency virus-positive patient; Regnery RL et al.; Isolation of a Rochalimaea-like organism from a febrile patient infected with human immunodeficiency virus was confirmed . Analysis of 16S rRNA gene sequences, together with polymerase chain reaction and restriction endonuclease length polymorphism analysis of a portion of the citrate synthase gene, demonstrated that the agent is closely related to members of the genus Rochalimaea and that the isolate is genotypically identical to the presumptive etiologic agent of bacillary angiomatosis . However, the same genotypic analyses readily differentiated the new isolate from isolates of other recognized Rochalimaea species as well as other genera of bacteria previously suggested as putative etiologic agents of bacillary angiomatosis and related syndromes . We propose that the novel species be referred to as Rochalimaea henselae sp . now. Plant Mol Biol, 1992 Feb, 18(3), 557 - 66 Cloning of a cDNA for rape chloroplast 3-isopropylmalate dehydrogenase by genetic complementation in yeast; Ellerstrom M et al.; Both insect and mammalian genes have previously been cloned by genetic complementation in yeast . In the present report, we show that the method can be applied also to plants . Thus, we have cloned a rape cDNA for 3-isopropylmalate dehydrogenase (IMDH) by complementation of a yeast leu2 mutation . The cDNA encodes a 52 kDA protein which has a putative chloroplast transit peptide . The in vitro made protein is imported into chloroplasts, concomitantly with a proteolytic cleavage . We conclude that the rape cDNA encodes a chloroplast IMDH . However, Southern analysis revealed that the corresponding gene is nuclear . In a comparison of IMDH sequences from various species, we found that the rape IMDH is more similar to bacterial than to eukaryotic proteins . This suggests that the rape gene could be of chloroplast origin, but has moved to the nucleus during evolution. J Cell Biol, 1992 Feb, 116(4), 923 - 31 Identification and characterization of an actin-binding site of CapZ; Hug C et al.; A mAb (1E5) that binds the COOH-terminal region of the beta subunit of chicken CapZ inhibits the ability of CapZ to bind the barbed ends of actin filaments and nucleate actin polymerization . CapZ prepared as fusion proteins in bacteria or nonfusion proteins by in vitro translation has activity similar to that of CapZ purified from muscle . Deletion of the COOH-terminus of the beta subunit of CapZ leads to a loss of CapZ's ability to bind the barbed ends of actin filaments . A peptide corresponding to the COOH-terminal region of CapZ beta, expressed as a fusion protein, binds actin monomers . The mAb 1E5 also inhibits the binding of this peptide to actin . These results suggest that the COOH-terminal region of the beta subunit of CapZ is an actin-binding site . The primary structure of this region is not similar to that of potential actin-binding sites identified in other proteins . In addition, the primary structure of this region is not conserved across species. Mol Cell Biol, 1992 Feb, 12(2), 609 - 18 Point mutations in the abl SH2 domain coordinately impair phosphotyrosine binding in vitro and transforming activity in vivo; Mayer BJ et al.; We have constructed a series of point mutations in the highly conserved FLVRES motif of the src homology 2 (SH2) domain of the abl tyrosine kinase . Mutant SH2 domains were expressed in bacteria, and their ability to bind to tyrosine-phosphorylated proteins was examined in vitro . Three mutants were greatly reduced in their ability to bind both phosphotyrosine itself and tyrosine-phosphorylated cellular proteins . All of the mutants that retained activity bound to the same set of tyrosine-phosphorylated proteins as did the wild type, suggesting that binding specificity was unaffected . These results implicate the FLVRES motif in direct binding to phosphotyrosine . When the mutant SH2 domains were inserted into an activated abl kinase and expressed in murine fibroblasts, decreased in vitro phosphotyrosine binding correlated with decreased transforming ability . This finding implies that SH2-phosphotyrosine interactions are involved in transmission of positive growth signals by the nonreceptor tyrosine kinases, most likely via the assembly of multiprotein complexes with other tyrosine-phosphorylated proteins. J Biotechnol, 1992 Feb, 22(3), 227 - 44 Some recent developments in the use of enzyme catalysed reactions in organic synthesis; Roberts SM et al.; The following processes are discussed in this article: enzyme-catalysed hydrolyses of carboxylic acid esters and amides, phosphate esters, nitriles and epoxides; esterification and inter-esterification reactions catalysed by enzymes; reduction of ketones to secondary alcohols using whole-cell systems or isolated dehydrogenases; oxidation of alicyclic and aromatic substrates using mono-oxygenases and dioxygenases in bacteria and fungi including enzyme-catalysed Baeyer-Villiger oxidations; aldol reactions, formation of optically active cyanohydrins and enzyme-catalysed acyloin type reactions . The use of these biocatalytic methods for the stereo-controlled preparation of important target structures is reviewed and some of the future directions for the biotransformation area are discussed. Zhonghua Wai Ke Za Zhi, 1992 Feb, 30(2), 81 - 3, 124 {Duodeno-biliary reflux during T-tube drainage: a clinical study}; Li XQ; The relationship between duodeno-biliary reflux and bile bacterial contamination was studied in 28 consecutive cases of T-tube drainage after cholecystectomy and choledochotomy . Patients were randomly divided into two groups of 14 each . Group I treated with positive pressure drainage (PPD), with the pressure in T-tube 0.3-0.5 kPa higher than that created by sphincter resistance . Group II treated with traditional negative pressure drainage (NPD) . It was found that the amount of duodenal reflux in group II was obviously larger than that in group I . We conclude that the traditional T-tube drainage incurs inevitably contamination of bacteria to bile by way of duodenal reflux . It is the consequence of bile flow dynamic disorder . PPD raises the bypass outflow resistance against the duodenal reflux and is thus superior to NPD. Rev Med Chil, 1992 Feb, 120(2), 197 - 201 {Classification of septicemias . A clinical and pathogenic approach}; Kraljevic R; Classic sepsis is characterized by the presence of bacteria in blood originating from a primary infection site with secondary location at other sites . Some infectious diseases like typhoid and paratyphoid fever, brucellosis an others share this pathogenetic mechanism but have a characteristic clinical course and usually a good prognosis . After analyzing the differences between the 2 types the author proposes the terms "non specific" and "specific" for each type of sepsis, respectively . The differences between the 2, the organism involved and different reaction of the host in types, may be related to different pathogenetic effects of each case. J Clin Periodontol, 1992 Feb, 19(2), 135 - 42 Enhanced gingivitis in the deciduous and permanent dentition . An experimental study in the dog; Berglundh T et al.; The aim of the experiment was to analyze the reaction of the marginal gingival tissues to 21 days of plaque formation on buccal tooth surfaces in the deciduous and permanent dentition of beagle dogs . In order to enhance the formation of plaque, the buccal surfaces of the experimental teeth were coated with a composite filling material . 5 beagle dogs were used . The animals were monitored during 2 periods, called period A (42 days during the deciduous dentition) and period B (42 days during the permanent dentition) . The dogs were 10 weeks old at the initiation of period A . Following 3 weeks of plaque control, a groove was prepared into the enamel of the buccal surfaces of the mandibular right 3rd (03P) and 2nd (02P) premolars . A cotton ligature was subsequently attached to the groove using an enamel/etch-technique and a composite filling material . The groove and the ligature did not interfere with the gingival margin but the composite material extended into the subgingival niche . The plaque control measures were abandoned . The animals formed plaque during the following 21 days . A clinical examination was performed and subgingival bacteria sampled on day 21 . Moreover, biopsies were harvested from the 03P and 02P tooth regions . The biopsies were prepared for histometric and morphometric analyses . A 2nd plaque control regimen was initiated . Period B started when the dogs were 15 months old . Following 3 weeks of enhanced plaque control, a cotton ligature was attached as described above at the buccal surfaces of the mandibular left 3rd (P3) and 4th (P4) premolars.(ABSTRACT TRUNCATED AT 250 WORDS) J Periodontol, 1992 Feb, 63(2), 80 - 6 Humoral antibody responses in periodontal disease; Zafiropoulos GG et al.; Several forms of periodontal disease are considered to be infectious diseases with associated specific bacteria . This study examined the humoral antibody levels as assayed by ELISA to Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia in adult periodontitis (AP), localized juvenile periodontitis (LJP), rapidly progressive periodontitis (RPP), and in periodontally healthy subjects (HS) . Sixty-two of the 64 (96.9%) patients had significantly elevated antibody levels to at least one of the three organisms . Elevated levels of antibodies to P . gingivalis occurred in 82.8% of the RPP, LJP, and AP patients with all 3 disease groups showing greater responses than HS controls . Antibodies to A . actinomycetemcomitans were found in 59.4% of the RPP, LJP, and AP patients and were significantly higher in both LJP and RPP patients . Only 21.9% of the RPP, LJP, and AP patients showed elevated levels to P . intermedia with only significantly higher levels in the RPP and LJP groups . Antibodies to A . actinomycetemcomitans and P . intermedia were rarely found alone (only 5.1% and 2.6% of the patients respectively) but were usually accompanied by P . gingivalis . These results suggest that one or more combinations of these 3 bacteria may play a role in the pathogenesis of these forms of periodontal disease. J Allergy Clin Immunol, 1992 Feb, 89(2), 545 - 51 Monocytes from patients with atopic dermatitis are primed for superoxide production; Polla BS et al.; To determine the potential role of monocytes (peripheral blood monocytes, PBMs) in the pathogenesis of atopic dermatitis (AD), we investigated whether PBMs from patients with severe AD are primed to generate toxic oxygen metabolites . To induce in vitro superoxide anion (O2-) production, we used either particulate (e.g., opsonized bacteria or zymosan) or soluble (e.g., phorbol esters) stimuli, which allowed us to test two distinct pathways for reduced nicotinamide-adenine dinucleotide phosphate activation . In addition, PBMs from the same patients were also examined for their levels of expression of the low-affinity receptor for IgE, Fc epsilon receptor 2 (CD23) . We found that PBMs, but not peripheral blood neutrophils, from patients with AD were primed for O2- production as compared to PBMs from either normal control subjects or patients with allergic rhinitis . These cells also expressed increased levels of CD23, and there was a significant correlation between these two parameters . Since the monocytes infiltrating into the AD lesions bear CD23, our data suggest that in vivo priming of PBMs and increased O2- production may participate in the pathogenesis of this skin disease. Artif Organs, 1992 Feb, 16(1), 19 - 42 Is a totally implantable artificial heart realistic? Nose Y. The incidence of local infection that occurred after the implantation of the intrathoracic Jarvik 7 total artificial heart (J-7 TAH) in patients was a major problem of this cardiac prosthesis . Infection rates increased when the J-7 TAH was implanted for longer periods of time, which was contrary to the results that were obtained by the implantation of the left ventricular assist devices (LVAD) either in the abdominal cavity or in the abdominal wall . Currently, the general belief is that the implantation of a cardiac prosthesis inside of the chest cavity is not safe, due to higher rates of infection . Thus, Part I of this paper deals with the question: "Is a total artificial heart physiologically acceptable?" We believe structural differences of the J-7 TAHs and other LVADs are an important part of the problem . The J-7 TAH is a volumetrically dynamic pump, and the other LVADs are a volumetrically fixed stationary pump . Based upon experiences of this investigator, intrathoracic implantation of a smooth surface pulsating device (or a volumetrically dynamic pump) generated persistent local inflammatory reactive tissues directly adjacent to the device . These tissue capsules were nonadherent to the device, and therefore may produce an ideal environment for allowing bacteria to grow . The findings were opposite to those for a non-pulsating surface pump inside the chest cavity . Part II of this paper discusses the question: "Is a totally implantable artificial heart technically achievable?" Many technical problems for the totally implantable TAH were already resolved during the development program of the totally implantable LVAD . However, there are three TAH specific problems that remain to be solved before achieving a clinically useful totally implantable TAH system . They are (a) anatomical compatibilities of the TAH system; (b) reliable control of the TAH system, (c) reliable and effective long-term operation of the TAH system . We have resolved the anatomical problem by the integral design of the pump-actuator system . Overall size of the hard shell Baylor TAH system is 510 cm3 with a diameter of 97 mm and a width of 82 mm . Its stoke volume was reduced to 63 cm3 . The stable and reliable control of TAH performances was established by the three sets of Hall effect sensors with left master alternate mode of pumping . No physiological parameter was used as a feedback signal . Reliable long-term operation of the TAH system was established by the electromechanical actuation system . A simple drive mechanism with commercially available components and subsystems was used.(ABSTRACT TRUNCATED AT 400 WORDS) J Biol Chem, 1992 Jan 25, 267(3), 1719 - 26 Contribution to ligand binding by multiple carbohydrate-recognition domains in the macrophage mannose receptor; Taylor ME et al.; The extracellular portion of the macrophage mannose receptor is composed of several cysteine-rich domains, including a fibronectin type II repeat and eight segments related in sequence to Ca(2+)-dependent carbohydrate-recognition domains (CRDs) of animal lectins . Expression of portions of the receptor in vitro, in fibroblasts and in bacteria, has been used to determine which of the extracellular domains are involved in binding and endocytosis of ligand . The NH2-terminal cysteine-rich domain and the fibronectin type II repeat are not necessary for endocytosis of mannose-terminated glycoproteins . CRDs 1-3 have at most very weak affinity for carbohydrate, so the carbohydrate binding activity of the receptor resides in CRDs 4-8 . CRD 4 shows the highest affinity binding and has multispecificity for a variety of monosaccharides . However, CRD 4 alone cannot account for the binding of the receptor to glycoproteins . At least 3 CRDs (4, 5, and 7) are required for high affinity binding and endocytosis of multivalent glycoconjugates . In this respect, the mannose receptor is like other carbohydrate-binding proteins, in which several CRDs, each with weak affinity for single sugars, are clustered to achieve high affinity binding to oligosaccharides . In the mannose receptor, these multiple weak interactions are achieved through several active CRDs in a single polypeptide chain rather than by oligomerization of polypeptides each containing a single CRD. J Immunol, 1992 Jan 15, 148(2), 584 - 9 Differing macrophage and lymphocyte roles in resistance to Legionella pneumophila infection; Yamamoto Y et al.; Similar to guinea pig macrophages and human monocytes, macrophages from the peritoneal cavity of thioglycolate pretreated A/J mice are permissive for growth of Legionella pneumophila . In contrast, macrophages from BDF1 mice are not permissive for L . pneumophila . Lymphocytes from A/J and BDF1 mice proliferated in response to Legionella Ag but guinea pig lymphocytes did not . Also, splenocyte cultures from A/J mice treated with either Con A or Legionella vaccine produced supernatants which induced A/J macrophages to restrict Legionella growth, but guinea pig splenocyte culture supernatants obtained after stimulation with L . pneumophila vaccine did not induce Legionella growth restriction activity by guinea pig macrophages . Murine rIFN-gamma but not rIFN-alpha markedly inhibited growth of Legionella in A/J mouse macrophages and monoclonal anti-IFN-gamma antibody neutralized the anti-Legionella activity of culture supernatants from A/J mouse splenocytes responding to Legionella Ag . From these data, IFN-gamma appears to be an important factor in anti-Legionella activity of Ag-activated mouse splenocyte culture supernatants . Cyclosporin A, when given to either A/J or BDF1 mice, reduced the proliferation responses of splenocytes to T cell mitogens and also decreased the IFN production of A/J spleen cells to Legionella Ag . In addition, drug treatment decreased the resistance of A/J mice to Legionella infection as shown by an increase in the number of viable bacteria in the liver . However, injection of drug treated mice with lymphokine-rich splenocyte culture supernatant reconstituted the resistance of these animals . These results suggest an important role for lymphocyte activation and lymphokine production in the resistance of A/J mice to Legionella infection . The greater resistance of BDF1 mice, however, may result from nonpermissive macrophages and responsive lymphocytes . In the case of guinea pigs, susceptibility to Legionella infections may result from both the permissive nature of the macrophages and the relatively unresponsive nature of the lymphocytes in these animals. Gene, 1992 Jan 15, 110(2), 239 - 43 Identification of a human cDNA with high homology to yeast omnipotent suppressor 45; Grenett HE et al.; Omnipotent suppression is a well-established phenomenon in yeast and bacteria in which nonsense mutations are misread . Wild-type (wt) suppressors are presumed to be involved in ensuring the fidelity of translation . We report a human homolog to wt yeast omnipotent suppressor 45 which shares 63% identity at the nucleotide level in the area of open reading frame (ORF) and 73% similarity at the amino acid (aa) level . The aa sequence of the human protein was deduced from a 2.3-kb cDNA (TB3-1) isolated from an adenocarcinoma T84 cell line cDNA library . The cDNA contains an ORF of 1284 bp which encodes a 47.8-kDa protein . Two transcripts for the clone were identified (2.6 and 4.0 kb) in a variety of human cell types . The strong structural similarity to yeast omnipotent suppressor 45, and its widespread expression suggest that this cDNA may play a role in the accurate recognition of nonsense codons in mammalian cells. FEMS Microbiol Lett, 1992 Jan 15, 69(3), 267 - 74 Detection of Leptospiraceae by amplification of 16S ribosomal DNA; Hookey JV; The polymerase chain reaction (PCR) was developed to detect Leptospiraceae . Primers were used to amplify 1 631 base-pair (bp) 5'-region of 16S rDNA . Representative strains from the species, Leptospira interrogans sensu stricto, L . borgpetersenii, L . noguchii, L . santarosai, L . weilii, L . inadai, L . meyeri and the single member strain of Leptonema were amplified . In contrast, strains representing the saprophytic species . L . biflexa, L . wolbachii and L . parva were not amplified . There was no PCR product from 23 phylogenetically unrelated species of bacteria . As little as 10-1 pg of purified DNA and as few as 10-1 leptospires could be detected using the PCR analysis . Isolates of leptospires from clinical sources gave a positive PCR band, but those from surface waters did not. Cancer Epidemiol Biomarkers Prev, 1992 Jan-Feb, 1(2), 131 - 5 Polyamine contents in rectal and buccal mucosae in humans treated with oral difluoromethylornithine; Boyle JO et al.; Difluoromethylornithine (DFMO) is an investigational chemopreventive agent that inhibits ornithine decarboxylase (ODC) activity, lowers cellular polyamine concentrations, and decreases cell proliferation in vivo and in vitro . In five subjects we have compared the polyamine concentrations in rectal mucosal biopsies and in exfoliated buccal mucosal cells (EBM) before and after DFMO treatment to assess the suitability of EBM as an easily accessible marker tissue for DFMO suppression of polyamine synthesis in the rectal mucosa . One month of 3 g/m2/day of DFMO treatment caused a statistically significant decrease in putrescine and spermidine concentrations in rectal mucosa biopsy specimens but not in EBM samples . ODC activity in EBM was high (approximately 1 mumol/min/mg protein), resistant to DFMO inhibition (Ki = 4200 microM), dependent on GTP concentration (maximal at 0.1 mM), and was reduced concomitantly with bacterial concentration by antiseptic mouthwashing . Bacteria adherent to EBM were visible by electron microscopy . Forty bacterial colonies/ng protein were culturable from washed EBM samples . Oral bacteria preclude the use of EBM samples as a marker tissue of DFMO effect in the rectal mucosa, but oral DFMO therapy is effective in depleting polyamines in rectal mucosa. J Biol Chem, 1992 Jan 5, 267(1), 310 - 8 Bradyrhizobium japonicum nodD1 can be specifically induced by soybean flavonoids that do not induce the nodYABCSUIJ operon; Smit G et al.; Besides genistein and daidzein, which are active inducers of the nodYABCSUIJ operon in Bradyrhizobium japonicum, soybean seeds also excrete compounds that are not inducers of the nodYABCSUIJ genes but enhance induction of this operon in the presence of a suboptimal genistein concentration . This synergism was studied in detail, and specific compounds were identified in seed exudate which specifically induce the nodD1 gene but not the nodYABCSUIJ operon . Therefore, our current hypothesis is that the observed synergism is caused by a specific induction of nodD1 . The specific nodD1 inducers from soybean seed extract have been purified and characterized chemically . They appear to be derivatives of genistein, glycitein, and daidzein with glucose, malonyl, and acetyl groups attached . Both root and seed exudate appear to contain these compounds, with the seed being the major source . No hydrolysis of these compounds to their aglycone forms was detected in the presence of B . japonicum . A model for nod gene induction in B . japonicum is discussed. J Biol Chem, 1992 Jan 5, 267(1), 649 - 52 Cloning of a Na(+)- and Cl(-)-dependent betaine transporter that is regulated by hypertonicity; Yamauchi A et al.; Many hypertonic bacteria, plants, marine animals, and the mammalian renal medulla are protected from the deleterious effects of high intracellular concentrations of electrolytes by accumulating high concentrations of the nonperturbing osmolyte betaine . When kidney-derived Madin-Darby canine kidney (MDCK) cells are cultured in hypertonic medium, they accumulate betaine to 1,000 times its medium concentration . This results from induction by hypertonicity of high rates of betaine transport into cells . We have isolated a cDNA (BGT-1) encoding a renal betaine transporter by screening an MDCK cell cDNA library for expression of a betaine transporter in Xenopus oocytes . The cDNA encodes a single protein of 614 amino acids, with an estimated molecular weight of 69 kDa . The deduced amino acid sequence exhibits highly significant sequence and topographic similarity to brain gamma-amino-n-butyric acid (GABA) and noradrenaline transporters, suggesting that the renal BGT-1 is a member of the brain GABA/noradrenaline transporter gene family . Expression in oocytes indicates that the BGT-1 protein has both betaine and GABA transport activities that are Cl(-)- as well as Na(+)-dependent and functionally similar to betaine and GABA transport in MDCK cells . Northern hybridization indicates that transporter mRNA is localized to the kidney medulla and is induced in MDCK cells by hypertonicity. Biochim Biophys Acta, 1992 Jan 3, 1123(1), 33 - 40 Transport of poly-beta-hydroxybutyrate in human plasma; Reusch RN et al.; Poly-beta-hydroxybutyrate (PHB) is an amphiphilic lipid that has been found to be a ubiquitous component of the cellular membranes of bacteria, plants and animals . The distribution of PHB in human plasma was investigated using chemical and immunological methods . PHB concentrations proved highly variable; in a random group of 24 blood donors, total plasma PHB ranged from 0.60 to 18.2 mg/l, with a mean of 3.5 mg/l . In plasma separated by density gradient ultracentrifugation, lipoproteins carried 20-30% of total plasma PHB; 6-14% in the very low density lipoproteins (VLDL), 8-16% in the low density lipoproteins (LDL), and less than 3% in the high density lipoproteins (HDL) . The majority of plasma PHB (70-80%) was found in protein fractions of density greater than 1.22 g/ml . Western blot analysis of the high density fractions with anti-PHB F(ab')2 identified albumin as the major PHB-binding protein . The affinity of albumin for PHB was confirmed by in vitro studies which demonstrated transfer of 14C-PHB from chloroform into aqueous solutions of human and bovine serum albumins . PHB was less tightly bound to LDL than to other plasma components; the polymer could be isolated from LDL by extraction with chloroform, or by digestion with alkaline hypochlorite, but it could not similarly be recovered from VLDL or albumin . PHB in the LDL correlated positively with total plasma cholesterol and LDL cholesterol, and negatively with HDL cholesterol . The wide concentration range of PHB in plasma, its presence in VLDL and LDL and absence in HDL, coupled with its physical properties, suggest it may have important physiological effects. Oncogene, 1992 Jan, 7(1), 65 - 70 A divergent ets-related protein, elk-1, recognizes similar c-ets-1 proto-oncogene target sequences and acts as a transcriptional activator; Rao VN et al.; The ets oncogene superfamily consists of a family of sequence-specific DNA-binding proteins that activate transcription . We have previously identified two new members of the ets oncogene superfamily, namely elk-1 and elk-2 . In this report we show that the recombinant elk-1 protein expressed in bacteria, like the c-ets-1 proto-oncogene, binds in a sequence-specific manner to Moloney murine sarcoma virus long terminal repeat, E74 target sequences and the PEA3 motif (polyoma enhancer), but does not bind to PU box sequences . Thus analysis of the DNA-binding specificity of ets-related proteins supports the view that different members show similar DNA-binding specificity, which is a general feature of the homeobox proteins . Our data using the chloramphenicol acetyltransferase gene linked to a thymidine kinase promoter containing multimers of the elk-1 target sequence indicates that elk-1 functions as a transcriptional activator . Interestingly, although elk-1 is the most divergent of all the members of the ets gene family, it shows very close similarities with c-ets-1 in some of its sequence-specific DNA-binding specificities . Here, we propose a new function for the elk-1 gene to act as a transcriptional activator of retroviruses and DNA tumor viruses. J Acquir Immune Defic Syndr, 1992, 5(3), 313 - 5 Instability of HIV sequences in high copy number plasmids; Peden KW; Plasmid proviral molecular clones of the LAI isolate of HIV-1 and the ROD isolate of HIV-2 were originally prepared in moderate copy number plasmids derived from pBR322, which contains the colE1 origin of replication . In these plasmid vectors, the HIV sequences are stable to continuous passage in bacteria . However, when the colE1 origin was replaced by the mutant origin from pUC18, pGEM, or pBluescript plasmids, which replicates to much higher copy numbers in bacteria, then deletions of HIV sequences occurred even in RecA defective strains . Deletions occurred in two different media, at room temperature and 37 degrees C, and with or without plasmid amplification in the presence of chloramphenicol . These results raise a cautionary note when cloning immunodeficiency viral sequences into plasmid vectors containing a high copy number origin of replication. Int J Syst Bacteriol, 1992 Jan, 42(1), 74 - 8 Taxonomic study of the genus Brachybacterium: Brachybacterium nesterenkovii sp . nov; Gvozdyak OR et al.; A new species, Brachybacterium nesterenkovii, is proposed for a group of coryneform bacteria that were isolated from milk products . These organisms have morphological, biochemical, and chemotaxonomic characteristics that are peculiar to the genus Brachybacterium . In contrast to strains of the only previously described species of the genus, Brachybacterium faecium, the representatives of the new species lack glycine in their peptidoglycan, although the peptidoglycan is of the same general type, and have large amounts of rhamnose in their cells . The strains of B . nesterenkovii exhibit no serological relationship with strains of B . faecium; in a numerical phenotypic analysis the two species were easily separated and formed clear-cut clusters . DNA-DNA hybridization between the type strains of B . faecium and B . nesterenkovii showed a level of homology of 22% . Strain 35 (= IMV Ac-752) is the type strain of B . nesterenkovii. Mol Gen Genet, 1992 Jan, 231(2), 286 - 95 Cell-free transcription of the nifH1 gene of Methanococcus thermolithotrophicus indicates that promoters of archaeal nif genes share basic features with the methanogen consensus promoter; Gohl HP et al.; The nifH1 gene of Methanococcus thermolithotrophicus, which encodes the putative dinitrogenase reductase of an archaeon, was accurately transcribed in a homologous cell-free transcription system . Extracts of cells grown with N2 or ammonia as nitrogen source initiated transcription at the nifH1 promoter with similar efficiencies . We confirmed that cells grown under non-N2-fixing conditions do not contain significant amounts of nifH1-specific mRNA . The levels of cell-free transcription initiation at the nifH1 promoter were similar to those observed at a tRNA promoter . The DNA sequence from -40 to +5 relative to the initiator nucleotide of nifH1 mRNA contained all the information required for promoter activity . A mutational analysis of this section of DNA demonstrated that a TATA box at -25 and the TTGT motif (initiator element) at the transcription start site are essential for cell-free transcription . These elements are similar to the structural determinants of a known tRNA promoter of Methanococcus . Mutation of a sequence, showing homology to the bacterial NifA site, which overlaps the transcription start site, did not affect promoter activity . Hence, cell-free transcription of the Methanococcus nifH1 gene is independent of upstream activator elements and does not require alternate cis-acting sequences that differ from the methanogen consensus promoter . These findings suggest that the activation of nif promoters is brought about by fundamentally different mechanisms in Archaea and bacteria. Biotechniques, 1992 Jan, 12(1), 88 - 97 Gene conversion of immunoglobulin variable regions in mutagenesis cassettes by replacement PCR mutagenesis; Near RI; A technique, Replacement PCR Mutagenesis, was developed to replace one immunoglobulin variable region (V) in a M13 phage cassette with a different, homologous V . This allows the use of the same mutagenesis and subsequent expression vectors for many V regions or V segments . The method combines PCR of V fragments and in vitro mutagenesis . Primers homologous to 3' and 5' ends of both V regions initiate PCR synthesis of the V DNA fragment (donor) that will replace the V region (recipient) in M13 . Donor V PCR DNA may originate from mRNA, cloned V genes or genomic templates . The donor V PCR DNA is denatured and annealed to the M13 cassette containing the recipient V to be supplanted . The second strand is synthesized, transfected into bacteria and mutant plaques selected by hybridization . Since restriction sites in primers are not required, altered primer-encoded amino acids are avoided . Further, the PCR donor piece can be of any length if it shares homology with the recipient gene . This allows construction and expression of complete gene replacements and chimeras . This method is also applicable to V "humanization" and studying sets of homologous genes containing polymorphic or evolutionary disparities . The potential uses of the technique are discussed. Carcinogenesis, 1992 Jan, 13(1), 77 - 81 The role of macrophage-derived TNFa in the induction of sublethal tumor cell DNA damage; Fulton AM et al.; In previous studies we showed that tumor-associated macrophages isolated from murine mammary tumors are mutagenic to bacteria and mammalian cells and thus may contribute to tumor progression . We reported previously, and confirm here, that inflammatory macrophages induce DNA strand breaks in cultured mammary tumor cells co-incubated at a 1:1 ratio for 1 h . This activity is prevented by inhibitors of arachidonate metabolism or the removal of H2O2 with catalase . In the present study, we show that two antibodies to recombinant murine tumor necrosis factor alpha (rMuTNFa)--a hamster monoclonal antibody (TN3-19.12) and a rabbit polyclonal antibody (Genzyme)--partially protect tumor cells from DNA strand breaks induced by elicited but not resident peritoneal macrophages . Antibody protection was reversed upon the addition of excess exogenous rMuTNFa . Purified rMuTNFa alone was unable to induce DNA strand breaks in the absence of macrophages, indicating that TNFa is necessary but not sufficient to mediate damage . Tumor target cells were completely resistant to the cytotoxic effects of rMuTNFa in the absence of actinomycin D and relatively resistant (in comparison to WEHI 164 clone 13 cells) in its presence . The incomplete protection seen with either catalase or anti-TNF suggests that macrophage-released TNFa, in the presence of other factors, induces non-cytotoxic DNA effects in tumor cells. Rev Environ Contam Toxico