Gene, 1995 Apr 14, 156(1), 69 - 74 Sequence of the bphD gene encoding 2-hydroxy-6-oxo-(phenyl/chlorophenyl)hexa-2,4-dienoic acid (HOP/cPDA) hydrolase involved in the biphenyl/polychlorinated biphenyl degradation pathway in Comamonas testosteroni: evidence suggesting involvement of Ser112 in catalytic activity; Ahmad D et al.; The nucleotide sequence of bphD, encoding 2-hydroxy-6-oxo-(phenyl/chlorophenyl)hexa-2,4-dienoic acid hydrolase involved in the biphenyl/polychlorinated biphenyl degradation pathway of Comamonas testosteroni strain B-356, was determined . Comparison of the deduced amino-acid sequence with published sequences led to the identification of a 'lipase box', containing a consensus pentapeptide sequence GlyXaaSerXaaGly . This suggested that the mechanism of action of this enzyme may involve an Asp-Ser-His catalytic triad similar to that of classical lipases and serine hydrolases . Further biochemical and genetic evidence for the active-site involvement of Ser112 was obtained by showing that a semipurified enzyme was inhibited by PMSF, a classic inhibitor of serine hydrolases, and by site-directed Ser112-->Ala mutagenesis. J Biol Chem, 1995 Apr 14, 270(15), 8411 - 9 Mutations at Pro67 in the RecA protein P-loop motif differentially modify coprotease function and separate coprotease from recombination activities; Konola JT et al.; The functional significance of residues in the RecA protein P-loop motif was assessed by analyzing 100 unique mutants with single amino acid substitutions in this region . Comparison of the effects on the LexA coprotease and recombination activities shows that Pro67 is unique among these residues because only at this position did we find substitutions that caused differential effects on these functions . One mutant, Pro67-->Trp, displays high constitutive coprotease activity and a moderate inhibitory effect on recombination functions . Glu and Asp substitutions result in low level constitutive coprotease activity but dramatically reduce recombination activity . The purified Pro67-->Trp protein shows a completely relaxed specificity for NTP cofactors in LexA cleavage assays and can use shorter length oligonucleotides as cofactors for cleavage of lambda cI repressor than can wild type RecA . Interestingly, both the mutant protein and wild type RecA can use very short oligonucleotides, e.g . (dA)6 and (dT)6, as cofactors for LexA cleavage . We have also found two mutations at position 67, which are completely defective for LexA coprotease activity in vivo but still maintain recombinational DNA repair (Pro67-->Lys) and homologous recombination (Pro67-->Lys and Pro67-->Arg) activities . These findings show that the recombination activities of RecA are mutationally separable from the coprotease function and that Pro67 is located in a functionally important position in the RecA structure. Biochim Biophys Acta, 1995 Apr 12, 1235(1), 93 - 9 Transport of glycine and lysine on the chloride-dependent beta-alanine (B0,+) carrier in rabbit small intestine; Munck LK et al.; Transport of glycine, lysine and beta-alanine in rabbit, guinea pig and rat small intestine has been examined by measurements of the unidirectional influx across the brush border membrane of the intact epithelium . In rabbit distal ileum the chloride-dependent fraction of glycine transport, and all sodium- and chloride-dependent lysine transport is carried on the beta-alanine carrier . Lysine eliminates all saturable, sodium-independent transport of glycine . The saturable, sodium-dependent, and lysine resistant influx of glycine is characterized by a K1/2Gly of 60 mM . Glycine transport in the mid intestine of the guinea pig is chloride-independent and in the rat only a minute fraction may be chloride-dependent . These species do not possess an equivalent of the rabbit beta-alanine carrier . In conclusion, glycine transport in rabbit distal ileum is by the sodium-dependent carrier of neutral amino acids, by the sodium-independent lysine carrier, and by the sodium- and chloride-dependent beta-alanine carrier which closely resembles the B0,+ carrier described in mouse blastocysts . All sodium dependent lysine transport in rabbit distal ileum is by the chloride- and sodium-dependent beta-alanine carrier . It is proposed that the beta-alanine carrier in rabbit distal ileum be renamed the B0,+ carrier. Proc Natl Acad Sci U S A, 1995 Apr 11, 92(8), 3463 - 7 Molecular cloning and characterization of a cDNA encoding monoclonal nonspecific suppressor factor; Nakamura M et al.; The monoclonal nonspecific suppressor factor (MNSF) is a lymphokine product of a murine T-cell hybridoma that inhibits the generation of lipopolysaccharide-induced immunoglobulin-secreting cells in an antigen-nonspecific manner . A cDNA clone encoding MNSF beta (an isoform of MNSF) was isolated and expressed in bacteria . The sequence obtained is virtually identical to the Fau protein, a product of the ubiquitously expressed fau gene with unknown function . Northern blot analysis demonstrated a single, 0.6-kb transcript . Specific polyclonal antibodies against synthetic peptides corresponding to the deduced amino acid sequences were elicited in rabbits . Immunoprecipitation experiments with these antibodies showed that MNSF beta is released extracellularly in an aggregate form, albeit it lacks a signal peptide sequence . The anti-MNSF beta affinity eluate from the MNSF-producing murine hybridoma (E17) and concanavalin A-activated splenocyte culture supernatants inhibited the immunoglobulin production by lipopolysaccharide-activated splenocytes . Recombinant MNSF beta also showed a similar biologic activity . Thus, ubiquitin-like protein(s) may be involved in the regulation of the immune responses. Proc Natl Acad Sci U S A, 1995 Apr 11, 92(8), 3132 - 6 Repression by SSN6-TUP1 is directed by MIG1, a repressor/activator protein; Treitel MA et al.; The SSN6-TUP1 protein complex represses transcription of diversely regulated genes in the yeast Saccharomyces cerevisiae . Here we present evidence that MIG1, a zinc-finger protein in the EGR1/Zif268 family, recruits SSN6-TUP1 to glucose-repressed promoters . DNA-bound LexA-MIG1 represses transcription of a target gene in glucose-grown cells, and repression requires SSN6 and TUP1 . We also show that MIG1 and SSN6 fusion proteins interact in the two-hybrid system . Unexpectedly, we found that LexA-MIG1 activates transcription strongly in an ssn6 mutant and weakly in a tup1 mutant . Finally, LexA-MIG1 does not repress transcription in glucose-deprived cells, and MIG1 is differentially phosphorylated in response to glucose availability . We suggest a role for phosphorylation in regulating repression. Proc Natl Acad Sci U S A, 1995 Apr 11, 92(8), 3100 - 4 Stathmin interaction with a putative kinase and coiled-coil-forming protein domains; Maucuer A et al.; Stathmin is a ubiquitous, cytosolic 19-kDa protein, which is phosphorylated on up to four sites in response to many regulatory signals within cells . Its molecular characterization indicates a functional organization including an N-terminal regulatory domain that bears the phosphorylation sites, linked to a putative alpha-helical binding domain predicted to participate in coiled-coil, protein-protein interactions . We therefore proposed that stathmin may play the role of a relay integrating diverse intracellular regulatory pathways; its action on various target proteins would be a function of its combined phosphorylation state . To search for such target proteins, we used the two-hybrid screen in yeast, with stathmin as a "bait." We isolated and characterized four cDNAs encoding protein domains that interact with stathmin in vivo . One of the corresponding proteins was identified as BiP, a member of the hsp70 heat-shock protein family . Another is a previously unidentified, putative serine/threonine kinase, KIS, which might be regulated by stathmin or, more likely, be part of the kinases controlling its phosphorylation state . Finally, two clones code for subdomains of two proteins, CC1 and CC2, predicted to form alpha-helices participating in coiled-coil interacting structures . Their isolation by interaction screening further supports our model for the regulatory function of stathmin through coiled-coil interactions with diverse downstream targets via its presumed alpha-helical binding domain . The molecular and biological characterization of KIS, CC1, and CC2 proteins will give further insights into the molecular functions and mechanisms of action of stathmin as a relay of integrated intracellular regulatory pathways. Mol Gen Genet, 1995 Apr 10, 247(1), 27 - 38 The cycHJKL gene cluster plays an essential role in the biogenesis of c-type cytochromes in Bradyrhizobium japonicum; Ritz D et al.; We present an extended genetic analysis of the previously identified cycH locus in Bradyrhizobium japonicum . Three new open reading frames found in an operon-like structure immediately adjacent to the 3' end of cycH were termed cycJ, cycK and cycL . A deletion mutant (delta cycHJKL) and biochemical analysis of its phenotype showed that the genes of the cluster are essential for the biogenesis of cellular c-type cytochromes . Mutations in discrete regions of each of the genes were also constructed and shown to affect anaerobic respiration with nitrate and the ability to elicit an effective symbiosis with soybean, both phenotypes being a consequence of defects in cytochrome c formation . The CycK and CycL proteins share up to 53% identity in amino acid sequence with the Rhodobacter capsulatus Cc11 and Cc12 proteins, respectively, which have been shown previously to be essential for cytochrome c biogenesis, whereas cycJ codes for a novel protein of 169 amino acids with an M(r) of 17857 . Localisation studies revealed that CycJ is located in the periplasmic space; it is probably anchored to the cytoplasmic membrane via an N-terminal hydrophobic domain . Based on several considerations discussed here, we suggest that the proteins encoded by the cycHJKL-cluster may be part of a cytochrome c-haem lyase complex whose active site faces the periplasm. Nurs Times, 1995 Apr 26-May 2, 91(17), 42 - 5 Hand decontamination: nurses' opinions and practices; Gould D; Infection is spread in hospital mainly by hands, making hand decontamination the most important means of preventing dissemination . There is some evidence to suggest that when access to hand-decontaminating agents is poor or the agents available are disliked, hands are washed too seldom, increasing risks of cross-infection . However, little attention has been paid to the use of towels and factors which promote their use, although it is known that damp hands transfer bacteria more readily than dry ones and that hands which become sore through poor drying have higher bacterial counts, contributing to the risk of cross-infection . This paper reports the results of the Nursing Times Hand Drying survey designed to assess nurses' access to hand decontamination agents and towels . The results suggest that the 112 nurses who participated were aware of the need for attention to hand hygiene but that access to both hand-decontaminating agents and paper towels was variable . Forty-one per cent complained of a shortage of soap and although nearly all used paper towels, these were in many cases of poor quality . Such towels were perceived as damaging to hands, leaving them feeling damp and sore . Good-quality, soft, paper towels were much appreciated by respondents in this sample . It is concluded that the quality of paper towels contributes to good infection control practice. J Gen Virol, 1995 Apr, 76 ( Pt 4), 1051 - 5 Expression of adenovirus type 5 E4 Orf2 protein during lytic infection; Dix I et al.; The human adenovirus type 5 E4 transcription unit has the potential to encode at least seven distinct polypeptides from reading frames accessed by differential splicing of a single primary transcript . Only some of these polypeptides have yet been detected during viral infection of cultured cells . Mutational inactivation of the reading frames whose products have not been described has no apparent effect on the growth of virus in standard cultured human cell lines, indicating that these proteins, if they exist, have only a subtle, non-essential role in the replication cycle . We have raised an antiserum to one of these undefined products, E4 Orf2, expressed in bacteria . Using this reagent, it was possible to show that Orf2 was expressed during the lytic cycle in HeLa cells, being a soluble cytoplasmic component appearing with early kinetics . No association of Orf2 protein with other infected cell components was detected. Indian J Pathol Microbiol, 1995 Apr, 38(2), 147 - 51 Study of bacteraemia using conventional and biphasic culture methods; Bannur M et al.; A total of two hundred blood samples collected from patients presenting with febrile illness were followed by culture using traditional and biphasic culture methods . Use of biphasic medium was observed to be of dual advantage as it grew pathogens earlier (28.5 percent in 18 hours, 63 percent in 36 hours of incubation) as well as in more number of cultures i.e . higher isolation rate (78 percent) when compared to traditional method. G E N, 1995 Apr-Jun, 49(2), 145 - 8 {Use of the Clotest and Pyloriset in the identification of Helicobacter pylori in asymptomatic healthy adults}; Santiago S; The Helicobacter Pylori (Hp) has been related with the pathogenesis of the chronic gastritis, peptic ulcer, non ulcerous dyspepsia, and in the last studies it has been found in a great prevailing way in populations with gastric cancer risks . The purpose of this study was to determine the presence of Hp in asymptomatic adults, volunteers, healthy, by means of the utilization of two diacritic tests, a serologic test for the determination of antibody IgG anti Hp (Pyloriset) and the tests Urease (Clotest) relating it with the development of lesions . We studied 20 persons, from whom we took peripheral blood samples for the determination of Hp by means of the agglutination test of latex (Pyloriset) and a superior digestive endoscopy was carried out (SDE) with a gastric biopsy for the Tests of Ureasa (Clotest) . In 15 adults (75%) the clotest was positive and in 13 persons (65%) the pyloriset was positive . In 9 persons (47%) the Hp was evidence by both methods . In 13 (65%) the SDE was pathological concluded and all were positive for the Hp . In 7 (35%) the SDE resulted normal . The presence of the antibody against the, Hp predicts alterations of the gastric mucosa like the development of the chronic gastritis, peptic ulcer and other pathologies . The Clotest is a simple tests, quick and sensible that allows us to apply immediate treatment . Our results suggest a high incidence of this bacteria in our workers, which justifies high prospective studies to determine same. Arq Gastroenterol, 1995 Apr-Jun, 32(2), 79 - 84 {Immunologic changes in alcoholic liver cirrhosis}; Calamita Z et al.; The alcoholic liver cirrhosis usually causes overall immunological changes which might be attributed to either the hepatic disease itself, to ethanol action and/or to malnourishment of the patient . These immune abnormalities comprise both cellular and humoral immunity, consisting of increased immunoglobulin levels, depressed late-skin response to antigens, lowered proliferative response of lymphocytes to mitogens, lower plasma levels of complement proteins (C3 and C4) and by either lower (IL2 and gamma IF) or increased (IL1, TNF, IL6 and IL8) cytokine levels . Parallel to the systemic immune suppression found in most patients, there is also a concomitant local, genetically based, immune stimulation at the liver level which leads to hepatic self-aggression . The systemic immune-suppression could be responsible for periodical infections or neoplasia found in these patients . The possible factors for the immune exhaustion are: a) lower hepatic clearance of toxins and/or bacteria; b) lower hepatic synthesis of complement components; c) cytokines (IL2 and gamma IF) deficiencies, and d) deficiencies of nutrients related to the antioxidant and/or immune defense mechanisms . The immune stimulation of the liver self aggression is characterized by the preferential migration of cytotoxic T cell and neutrophils to the liver, following stimulatory factors such as Mallory bodies, acetaldehyde and/or antibodies . Moreover, the local increase of cytokines (IL1, TNF, IL6 and IL8) levels would be liable for the local phagocyte chemotaxy (IL8) or part of liver injury (TNF) eased by the lower antioxidant defense of the cirrhotic liver.(ABSTRACT TRUNCATED AT 250 WORDS) Am J Clin Nutr, 1995 Apr, 61(4 Suppl), 915S - 921S Sugars: their origin in photosynthesis and subsequent biological interconversions; Nevins DJ; Sugar has been valued as a commodity for thousands of years . Despite its long history in commerce, the biological mechanisms accounting for the production of sugar are rather recent discoveries . The reactions are remarkable . Sugar is produced by all green plants and photosynthetic bacteria in a reaction sequence capable of forming carbon-carbon bonds . The very first steps occur independently of solar energy input, but to sustain the reaction, the products of initial fixation are phosphorylated and undergo a reduction in oxidation state . These steps responsible for phosphorylation and reduction are driven by products generated in the chloroplast upon the absorption of light . At this point, after just a few reactions, the products of photosynthesis have already acquired the attributes characteristic of sugars . Once carbon is stabilized as simple sugars in the chloroplast, the products undergo a sequence of rearrangements to sustain a cycle leading to new carbon dioxide acceptor molecules, and with each turn of the cycle a new carbon atom is introduced into the pool . As the process continues some of the carbon is diverted to synthesize starch within the chloroplast . Sucrose is synthesized in the cytoplasm adjacent to the chloroplast from exported carbohydrate as a diversion from the formation of starch . Sucrose represents the principal transport substance in most plants . Storage starch, cellulose, and other complex cell wall polysaccharides are typically derived from the sugar monomers found in sucrose . Sugars supply all the fixed carbon for synthesis of biological compounds and are fundamental for sustaining the energy flow to all food systems. J Am Acad Dermatol, 1995 Apr, 32(4), 640 - 52 Antigen processing: the gateway to the immune response; Kalish RS; The T-lymphocyte response to an antigen is governed by the source of that antigen and the way in which it is processed . Before recognition by T lymphocytes, proteins must be degraded to peptides by antigen-presenting cells . The peptides are then presented on major histocompatibility complex (MHC) molecules for recognition by the T cells . Antigens arising outside the cell (e.g., bacteria) are phagocytosed and processed by the exogenous pathway for presentation on MHC class II molecules (e.g., DR) to CD4+ cells . Antigens derived from the cytoplasm (e.g., viral proteins) are processed by the endogenous pathway for presentation by MHC class I molecules (e.g., HLA-A, -B, -C) to CD8+ cells . The response to a hapten or drug is a function of the antigen processing pathway and is determined by its chemical properties . Antigen processing also governs the T-cell response to pathogens, vaccines, and autoimmune conditions. J Bacteriol, 1995 Apr, 177(7), 1760 - 5 Overlapping promoters for two different RNA polymerase holoenzymes control Bradyrhizobium japonicum nifA expression; Barrios H et al.; The Bradyrhizobium japonicum NifA protein, the central regulator for nitrogen fixation gene expression, is encoded in the fixRnifA operon . This operon is activated during free-living anaerobic growth and in the symbiotic root nodule bacteroid state . In addition, it is expressed in aerobic conditions, albeit at a low level . Here, we report that this pattern of expression is due to the presence of two overlapping promoters: fixRp1, which is of the -24/-12 class recognized by the RNA polymerase sigma 54, and fixRp2, which shares homology with the -35 and -10 regions found in other putative B . japonicum housekeeping promoters . Primer extension analyses showed that fixRp1 directed the synthesis of a transcript, P1, that starts 12 nucleotides downstream of the -12 region . In addition to sigma 54, P1 was dependent on NifA and low oxygen tension . Transcripts originating from fixRp2 started at two sites: one coincided with P1, while the most abundant, P2 initiated just two nucleotides further downstream of P1 . Expression from fixRp2 was dependent on the upstream -68 promoter region, a region known to bind a putative activator protein, but it was independent of sigma 54 and NifA . This promoter was expressed in aerobic and anaerobic conditions but was not expressed in 30-day-old bacteroids . Mutations in the conserved 12 region for the sigma 54 promoter did not show any transcript, because these mutations also disrupted the overlapping -10 region of the fixRp2 promoter . Conversely, mutations at the -24 region only affected the sigma 54-dependent P1 transcript, having no effect on the expression of P2 . In the absence of omega(54), anaerobic expression from the fixRp(2) promoter was enhanced threefold, suggesting that in the wild-type strain, the two RNA polymerase holoenzymes must compete for binding to the same promoter region. Infect Immun, 1995 Apr, 63(4), 1391 - 3 Deviation of immune response to Chlamydia psittaci outer membrane protein in lipopolysaccharide-hyporesponsive mice; Westbay TD et al.; The outcome of infection is determined by both the quantity and the quality of an induced immune response . In particular, it has been demonstrated for selected pathogens that induction of TH1 or TH2 type helper T-cell subsets determines whether an immune response gives rise to protective immunity or disease-associated immunopathology . The nature of the antigen and the type of antigen-presenting cells recruited in the induction of a response are critical factors that influence the quality of the immune response . Of particular interest in this respect is the immune response to bacterial particles and the impact of cell wall-associated lipopolysaccharide (LPS) on that response . Nonspecific activation of macrophages and B lymphocytes by LPS could skew the phenotype of activated antigen-presenting cells and selectively alter the immunoglobulin isotypes and helper T-cell subsets that are induced following infection . In an initial attempt to detect immune deviation associated with LPS stimulation, we have compared the immunoglobulin isotypes of antibodies specific for the cysteine-rich outer membrane protein Omp2 induced in normal and LPS-hyporesponsive mice following immunization with Chlamydia psittaci strain guinea pig inclusion conjunctivitis whole elementary bodies . We report that there is a dramatic shift of Omp2-specific antibody from predominantly immunoglobulin G2a (IgG2a) isotype in LPS-hyporesponsive mice to high levels of IgG1 isotype in LPS-responder strains . The dependence of the IgG1 isotype shift on the LPS responder status is linked to the structure of the antigen and its natural processing pathway since LPS-hyporesponsive mice are not, in general, deficient in IgG1 antibody production . In particular, the antibody response to purified recombinant Omp2 is predominantly of the IgG1 isotype even in LPS-hyporesponsive mice. Surg Neurol, 1995 Apr, 43(4), 344 - 50 A no-touch technique protocol to diminish cerebrospinal fluid shunt infection; Faillace WJ; BACKGROUND: Cerebrospinal fluid (CSF) shunts become infected primarily by bacterial organisms indigenous to the patient's skin flora, but bacteria from the operating room environment, hospital, or no obvious source may also infect a shunt . To decrease the incidence of shunt infection, a no-touch technique protocol was developed and utilized in a prospective manner . METHODS: A before-after trial analysis was performed to compare the infection rates between patients who had CSF shunts placed using the no-touch technique protocol versus patients who had surgery without the protocol . Patients were stratified by age, etiology of hydrocephalus, type of shunt surgery, and presence of a contaminated skin wound, namely, tracheostomy, gastrostomy-jejunostomy, colostomy, or halo . The differences in infection rates were analyzed with the Fisher exact test with midpoint value correction, and standard statistical methods were used to calculate the 90% confidence interval odds ratio and number to treat . RESULTS: The no-touch technique protocol resulted in a clinically significant threefold decrease in shunt infection rate from 9.1% to 2.9% (p = 0.058 at 0.10 level, odds ratio 0.305, confidence interval 0.084-1.11), and a near threefold decrease in the infection rate per patient from 11.3% to 3.9% (p = 0.032 at 0.10 level, odds ratio 0.243, confidence interval 0.065-0.906) . CONCLUSIONS: The no-touch technique protocol as described herein is a useful method to decrease shunt infection . A larger prospective, randomized, multicenter clinical trial is encouraged to stringently assess the efficacy of the protocol. Poult Sci, 1995 Apr, 74(4), 742 - 52 The effects of ultrapasteurization with and without homogenization on the chemical, physical, and functional properties of aseptically packaged liquid whole egg; Martinez RM et al.; The effects of ultrapasteurization with and without homogenization on some chemical, physical, and functional properties of liquid whole egg were observed . Heat treatments of 64, 68, and 72 C each at 30, 60, and 95 s were conducted on liquid whole egg, unhomogenized and homogenized {10.34 mPa (1,500 psi)} . The viscosity of the egg increased with increased processing temperature, and unhomogenized egg was more viscous than homogenized egg . alpha-Amylase was inactivated at 68 C for 30 s . Although cake height was highly correlated to soluble protein content, this relationship did not hold for all time-temperature combinations . Due to homogenization effects on soluble protein results, soluble protein is not a reliable predictor of egg functionality when homogenization is used in the process . Homogenization had no effect on the population reduction of aerobic bacteria in all but three of the nine time-temperature combinations. Immunology, 1995 Apr, 84(4), 549 - 54 Production of TNF-alpha, IL-6 and TGF-beta, and expression of receptors for TNF-alpha and IL-6, during murine Mycobacterium avium infection; Champsi J et al.; The Mycobacterium avium complex comprises intracellular bacteria associated with disseminated infection in patients with acquired immune deficiency syndrome (AIDS) . Immune defects that lead to infection are unknown but cytokines appear to play an important role in the immunomodulation of host defence mechanisms . We evaluated the cytokine profiles seen temporally after murine M . avium infection . Spleen cells were obtained from M . avium-infected C57BL/6 mice and uninfected mice at weeks 1, 2, 3, 4 and 5 . Cells were cultured in vitro and subsequently pulsed with killed M . avium . Supernatants were collected from the cultured splenic cells and the concentrations of interleukin-6 (IL-6), transforming growth factor-beta 1 (TGF-beta 1) and tumour necrosis factor-alpha (TNF-alpha) were measured . TGF-beta 1 was detected at week 1, followed by IL-6 production at week 2 . Elevated TNF-alpha levels were observed at week 3 . The addition of polyclonal anti-TGF-beta 1 antibody to M . avium-infected peritoneal macrophages in the presence of splenic cell supernatants from weeks 1, 3 and 5 led to decreased bacterial counts compared to controls . Anti-IL-6 antibody did not have any effect on macrophage anti-mycobacterial activity . Concurrently, we observed decreased expression of TNF-alpha receptors on infected macrophages . We propose that the early elevated levels of TGF-beta 1, a known suppressor of macrophage function, in conjunction with down-regulation of TNF-alpha receptors may help explain the suboptimal macrophage response to TNF-alpha, leading to impaired anti-mycobacterial activity. Genetics, 1995 Apr, 139(4), 1469 - 81 Involvement of the host initiator function dnaA in the replication of coliphage lambda; Wegrzyn G et al.; We demonstrate that the initiation of coliphage lambda DNA replication is dependent on the host initiator function dnaA, provided that the lambdoid prophage Rac is absent . Presence of Rac compensated the absence of dnaA function, causing initiation of replication . In dnaAts rac+ cells at 43 degrees, most of parental phage DNA molecules, after one round of theta replication, switched to a replication with features of the sigma mode and produced progeny at high yield . Initiation of replication of the lambda Pts1 mutant at 43 degrees was blocked by dnaA function; however, under dnaA-rac+ conditions all parental phage DNA molecules, after one round of theta replication, switched to the sigma mode and produced progeny at high yield . Taking into account our recent finding that transcriptional activation of ori lambda seems to be dnaA-regulated (to be published elsewhere), we suggest that the DnaA-lambda Pts1 incompatibility occurs at the insertion of the ori lambda-bound lambda O-lambda P-DnaB preprimosome between the complementary lambda DNA strands . The role of Rac and the mechanism of the switch from theta to sigma mode of lambda phage DNA replication are discussed. J Periodontol, 1995 Apr, 66(4), 279 - 84 Modulation by progesterone of interleukin-6 production by gingival fibroblasts; Lapp CA et al.; The gingivitis associated with pregnancy has been attributed to increased concentrations of circulating estrogen and/or progesterone . However, the mechanism by which these steroids increase gingival inflammation is not known . Interleukin-6 (IL-6), a pleiotropic cytokine produced by many cell types including human gingival fibroblasts (hGF), is secreted in response to inflammatory challenges such as bacterial lipopolysaccharide and interleukin-1 (IL-1) . This study tested the hypothesis that progesterone could modulate the local production of IL-6 by hGF . The effects of progesterone on IL-6 production were measured in vitro in serum-free, phenol red-free medium to eliminate possible effects of such medium additives . The concentration of IL-6 secreted into supernatant medium after a 24 hour challenge with IL-1 beta was estimated by radioimmunoassay . Total RNA from steroid-treated hGF was probed for IL-6 mRNA . In serum-free medium, progesterone dose-dependently and significantly (P < 0.05) inhibited IL-6 production by hGF, as did the glucocorticoids hydrocortisone (HC) and dexamethasone . At progesterone concentrations common in late pregnancy, IL-6 production was reduced to levels 40 to 50% of control . In addition, mRNA was significantly down-regulated by progesterone and HC, at both basal levels and after IL-1 beta challenge . These results suggest that high levels of progesterone during pregnancy affect the development of localized inflammation by down-regulation of IL-6 production, rendering the gingiva less efficient at resisting the inflammatory challenges produced by bacteria. J Periodontol, 1995 Apr, 66(4), 274 - 8 Polyamines found in gingival fluid inhibit chemotaxis by human polymorphonuclear leukocytes in vitro; Walters JD et al.; Putrescine and spermidine occur at concentrations approaching 1 mM in gingival fluid at diseased periodontal sites . Previous work demonstrates that these polyamines potentiate Ca2+ signaling in polymorphonuclear leukocytes (PMNs), resulting in enhanced degranulation and superoxide generation . The present study extends this work by characterizing the effects of polyamines on PMN chemotaxis and phagocytosis, in which Ca2+ signaling plays a less defined regulatory role . Putrescine (1 mM) and spermidine (0.1 to 0.5 mM) significantly inhibited chemotaxis to fMet-Leu-Phe and C5a (P < 0.05) . This inhibition was not strongly related to any effect polyamines have on PMN adhesion, actin polymerization, or formyl peptide receptor expression . Neither putrescine nor spermidine had a significant impact on phagocytosis of opsonized bacteria by PMNs . Thus, at concentrations similar to those found in gingival fluid, polyamines could potentially inhibit recruitment of PMNs to diseased pockets without impairing their ability to engulf invading bacteria. J Laryngol Otol, 1995 Apr, 109(4), 300 - 3 The management of sinogenic orbital complications; Singh B; The place of conservative treatment in sinogenic orbital complications has not been fully explained in the literature . The question that remains unresolved is--at which stage of the disease is surgery indicated? A study was undertaken in 240 patients with sinogenic orbital complications, to determine this . The patients were divided into three groups according to the stage of the disease as determined clinically: Group 1 (52 patients) with early stage disease, as detected by cellulitis only; Group 2 (76 patients) with intermediate stage, as detected by periorbital cellulitis and proptosis, but with full range of eye movement and unaltered vision; Group 3 (122 patients) with late stage disease, as detected by periorbital cellulitis and gross proptosis, with limitation of eye movement and altered vision . Group 1 and Group 2 patients were treated conservatively, with intravenous antibiotics and antral lavage . Group 3 patients were treated with intravenous antibiotics and surgery . External frontoethmoidectomy was performed in 31 (bilateral in two), ethmoidectomy in 91 (bilateral in five), sphenoidectomy in 15 and bilateral antral washout in all (122 patients) . There was 100 per cent success with conservative treatment in Group 1 patients, whilst in Group 2 there was 98.6 per cent failure . The 75 patients in whom conservative treatment failed were successfully treated with surgery: frontoethmoidectomy was performed in 66 and ethmoidectomy in nine . In Group 3 patients, 100 per cent success was achieved with intravenous antibiotics and surgery . Sinogenic orbital complications can be treated conservatively and surgically, depending on the stage of the disease on presentation . Conservative treatment is only suitable for early complications, i.e . patients with periorbital cellulitis only.(ABSTRACT TRUNCATED AT 250 WORDS) Arzneimittelforschung, 1995 Apr, 45(4), 486 - 90 Studies on the in vitro and in vivo genotoxicity of {2,2-dimethyl-6- (4-chlorophenyl)-7-phenyl-2,3-dihydro-1H-pyrrolizine-5-yl}-acetic acid; Heidemann A et al.; {2,2-Dimethyl-6-(4-chlorophenyl)-7-phenyl-2,3-dihydro-1H-pyrrolizine-5- yl}-acetic acid (CAS 156897-06-2, ML 3000) was examined for genotoxic activity in bacteria and mammalian cells in vitro as well as in vivo . The substance did not increase gene mutation frequencies either in a bacterial system or in a cultured V79 cell line of the Chinese hamster . Both in vitro tests were conducted in the presence and absence of S9-mix . In the unscheduled DNA synthesis assay in vitro with primary rat hepatocytes, negative results were also obtained . A cytogenetic analysis of the bone marrow of male and female Wistar rats was performed . After oral application ML 3000 did not increase the number of cells with structural chromosomal aberrations . The results suggest that ML 3000 has no genotoxic potential in vitro and in vivo. J Mol Evol, 1995 Apr, 40(4), 455 - 63 Evolutionary analysis of aspartate aminotransferases; Winefield CS et al.; Aspartate aminotransferase isoenzymes are located in both the cytosol and organelles of eukaryotes, but all are encoded in the nuclear genome . In the work described here, a phylogenetic analysis was made of aspartate aminotransferases from plants, animals, yeast, and a number of bacteria . This analysis suggested that five distinct branches are present in the aspartate aminotransferase tree . Mitochondrial forms of the enzyme form one distinct group, bacterial aspartate aminotransferase formed another, and the plant and vertebrate cytosolic isoenzymes each formed a distinct group . Plant cytosolic isozymes formed a further group of which the plastid sequences were a member . The yeast mitochondrial and cytosolic aspartate aminotransferases formed groups separate from other members of the family. Eur J Oral Sci, 1995 Apr, 103(2 ( Pt 1)), 77 - 83 Spontaneous gingival antibody production to Fusobacterium nucleatum outer membrane in patients with adult periodontitis; Nunes IP et al.; The local antibody response to Fusobacterium nucleatum outer membrane (FnOM) was analyzed in patients with adult periodontitis (AP) at the single cell level . Furthermore, we analyzed whether periodontal hygienic treatment could alter the antibody response . The number of IgG- and IgM-producing cells were investigated in gingival samples collected from 20 patients with AP . The patients were divided into 2 groups, before (BT, n = 9) and after (AT, n = 11) periodontal hygienic treatment . Four healthy gingival samples were used as controls . The results obtained showed that local antibody production against FnOM occurred in gingiva of patients with AP, but not in healthy gingiva . The IgG anti-FnOM was the predominant isotype observed . However, there was no statistically significant difference between the BT and AT groups . These results indicate that periodontal hygienic treatment was not sufficient to alter significantly the number of IgG- and IgM-secreting cells present in gingival tissue of AP patients, but it promoted a reduction of IgG anti-FnOM secreting cells . The presence of anti-FnOM antibodies in AP but not in control patients indicates that this bacteria may play a role in the pathogenesis of periodontal disease. Curr Microbiol, 1995 Apr, 30(4), 247 - 50 Infectivity of Legionella pneumophila mip mutant for alveolar epithelial cells; Cianciotto NP et al.; Legionella pneumophila can invade and grow within explanted alveolar epithelial cells . Given its potential clinical significance, an examination of the molecular basis of epithelial cell infection was initiated . The mip gene encodes a 24-kilodalton surface protein that promotes macrophage infection and virulence . To determine whether this gene is required for pneumocyte infection, we tested a strain bearing a mip null mutation for its ability to infect both explanted type II cells and type I-like cell lines . For infection of type II cells, the infective dose 50% for the Mip-strain was 25-fold higher than an isogenic Mip+ strain . Type I cell monolayers infected with the mutant for 3 days yielded approximately 50-fold fewer bacteria than did monolayers infected with the parental strain . These data indicate that Mip enhances infection of pneumocytes and that L . pneumophila employs some of the same genes (mechanisms) to infect epithelial cells and macrophages. Kansenshogaku Zasshi, 1995 Apr, 69(4), 438 - 43 {An autopsy case of fungal (Mucor) cerebral aneurysm}; Tokuda T et al.; An autopsy of rupture of Mucor cerebral aneurysm, not diagnosed during the patient's life, was experienced . A 63-year-old female was admitted to our hospital with the chief complaint of disturbance of consciousness and high fever . Her past histories were diabetes mellitus, liver cirrhosis and nasal sinusitis . The remarkable findings on admission were moderate inflammatory data, high blood sugar level in serum and ascites . Brain CT film revealed a non-enhanced low-density area in the frontal region . The cerebrospinal fluid showed bloody color and white blood cell counts were 3300/microliter (mostly neutrophils) . Under our suspected of bacterial encephalomeningitis, intravenous cefotaxime and ampicillin therapy was started immediately . Cultures of cerebrospinal fluid for bacteria were negative . The disturbance of her consciousness gradually improved under general treatment . However, her conscious level suddenly became a coma on the 6th hospital day and she died on the 9th hospital day . An autopsy revealed Mucor at the site of the rupture of the cerebral aneurysm. Iowa Med, 1995 Apr, 85(4), 166 - 8 Beyond measles and influenza: the future of vaccines; Dordick V; The usefulness of antibiotics has become more limited due to the growing ability of bacteria to become resistant . The preventive potential of vaccines may offer solutions to some of today's medical challenges, say UI experts. FASEB J, 1995 Apr, 9(7), 504 - 8 Mercury exposure from "silver" tooth fillings: emerging evidence questions a traditional dental paradigm; Lorscheider FL et al.; For more than 160 years dentistry has used silver amalgam, which contains approximately 50% Hg metal, as the preferred tooth filling material . During the past decade medical research has demonstrated that this Hg is continuously released as vapor into mouth air; then it is inhaled, absorbed into body tissues, oxidized to ionic Hg, and finally covalently bound to cell proteins . Animal and human experiments demonstrate that the uptake, tissue distribution, and excretion of amalgam Hg is significant, and that dental amalgam is the major contributing source to Hg body burden in humans . Current research on the pathophysiological effects of amalgam Hg has focused upon the immune system, renal system, oral and intestinal bacteria, reproductive system, and the central nervous system . Research evidence does not support the notion of amalgam safety. Plast Reconstr Surg, 1995 Apr, 95(5), 860 - 5 Quantitative culture technique and infection in complex wounds of the extremities closed with free flaps; Breidenbach WC et al.; The purpose of this study was twofold: (1) to determine if there is a relationship between a critical quantity of bacteria and infection in complex extremity wounds, and (2) to compare the positive and negative predictive values, sensitivities, and specificities of the following laboratory and clinical tests of potential wound infections: quantitative cultures, swab cultures, mechanism of injury, severity of fracture, and wound position . Fifty patients were evaluated who had 50 free-tissue transfers . Twenty-eight patients had quantitative cultures prior to flap coverage . All 50 patients were evaluated for wound position, presence or absence of a fracture, and mechanism of injury; 16 patients had swab cultures . The best positive predictive value (89 percent) and negative predictive value (95 percent), along with sensitivity (89 percent) and specificity (95 percent), for predicting wound infection were those of quantitative cultures . This demonstrates that in the clinical setting of this study, the test with the highest utility and validity is quantitative cultures. J Infect Dis, 1995 Apr, 171(4), 924 - 9 Bordetella pertussis fimbriae bind to human monocytes via the minor fimbrial subunit FimD; Hazenbos WL et al.; Nonopsonized Bordetella pertussis can bind to and become ingested by human monocytes . Previous studies demonstrated that mutant B . pertussis strains that lack fimbriae express a reduced adherence to monocytes . The present study was undertaken to investigate the involvement of the minor fimbrial subunit FimD in the adherence of B . pertussis to human monocytes using purified fimbriae, FimD, and strains with mutations in fimbrial genes . Flow cytometry demonstrated that purified B . pertussis fimbriae avidly bind to monocytes in a dose-dependent manner but bind less to neutrophils and hardly to lymphocytes; FimD-lacking fimbriae did not bind to monocytes . Purified fimbriae or FimD inhibited the binding of wild-type B . pertussis to monocytes in a dose-dependent manner and similarly inhibited the binding of a mutant defective in the major fimbrial subunits . It did not affect the binding of strains defective in FimD . These results prove that B . pertussis bind to human monocytes via the fimbrial minor subunit FimD. J Infect Dis, 1995 Apr, 171(4), 1060 - 4 Muramic acid in peripheral blood leukocytes of healthy human subjects; Lehtonen L et al.; Peptidoglycan, a specific marker for all bacterial cell walls, was studied in peripheral blood of healthy human subjects by mass spectrometric analysis of muramic acid . Peripheral blood mononuclear and polymorphonuclear cells from 98 healthy adults were hydrolyzed and analyzed by gas chromatography-mass spectrometry as alditol acetate derivatives using selective ion monitoring . Muramic acid was observed in cell samples from 21 of 98 subjects . Blood cultures were done simultaneously and remained negative . As a control, mononuclear and polymorphonuclear cells separated from umbilical vein blood of 41 healthy newborns were studied; all were muramic acid-negative . Since newborns lack gut flora, intestinal absorption of bacteria or of their degradation products appears to be the most likely explanation for the finding. J Clin Invest, 1995 Apr, 95(4), 1690 - 701 Phenotypic and functional changes in peripheral blood monocytes during progression of human immunodeficiency virus infection . Effects of soluble immune complexes, cytokines, subcellular particulates from apoptotic cells, and HIV-1-encoded proteins on monocytes phagocytic function, oxidative burst, transendothelial migration, and cell surface phenotype; Trial J et al.; We postulated that changes in the cell surface display of molecules that facilitate cell-cell and cell-matrix adhesions may reflect the changing immunosurveillance capacity of blood monocytes during progression of human immunodeficiency virus (HIV) infections . In Centers for Disease Control (CDC) stage A patients, whose monocytes' ability to phagocytose bacteria and generate reactive oxygen intermediates is often increased, the frequency of monocytes expressing CD49d, HLA-DP, HLA-DQ, and an activation epitope of CD11a/CD18 was increased and monocyte transendothelial migration was unimpaired . In CDC stage B/C patients, whose monocytes' ability to phagocytose bacteria and migrate across confluent endothelial monolayers was diminished, surface expression of CD49e and CD62L and the percentage of monocytes expressing CD18, CD11a, CD29, CD49e, CD54, CD58, CD31, and HLA-I were significantly decreased . Incubating normal donor monocytes with immune complexes in vitro reproduced the phenotypic and functional abnormalities seen in stage B/C patients . By contrast, in vitro stimulation with subcellular particulates released by apoptotic lymphocytes reproduced changes seen in stage A patients' monocytes . Although circulating monocytes appear to be activated at all stages, these data suggest that the high levels of circulating immune complexes, found predominantly in the later stages of HIV infection, may be particularly instrumental in reducing the monocyte's capacity to maintain surveillance against infection. Exp Cell Res, 1995 Apr, 217(2), 517 - 27 Adhesion activity in fibronectin's alternatively spliced domain EDa (EIIIA): complementarity to plasma fibronectin functions; Xia P et al.; The EDa (EIIIA) domain is one of two alternatively spliced type III homology repeats in fibronectins which differentiate cellular (cFN) from plasma isoforms (pFN) . A bacteria-expressed recombinant polypeptide encoding the EDa type III repeat promotes adhesion of some cell types and its activity is synergistic with neighboring repeats III11 and III12 . We show in this study that co-coating substrata with the EDa-only polypeptide and a suboptimal concentration of pFN leads to increased attachment and extensive spreading of v-src-transformed 3T3 cells relative to that found on substrata of suboptimal pFN or EDa polypeptide alone . This complementarity of activities requires as little as 1 microgram/ml EDa polypeptide in the adsorbing mixture and displays sequence specificity for only EDa (recombinant polypeptides of neighboring repeats III11 or III12 were without effect) . Furthermore, stress fibers and focal contacts are inducible on the EDa:pFN mixture, suggesting that the EDa sequence and its receptor participate in signal transduction . The codistribution of phosphotyrosine proteins, including pp125FAK, along with vinculin and talin into focal contacts supports this hypothesis . Therefore, an alternatively spliced domain EDa which is expressed in various proportions in cells and tissues may have special functions related to adhesion processes by complementing the functions of pFN circulating in blood. Avian Dis, 1995 Apr-Jun, 39(2), 360 - 7 Bordetella avium: an opportunistic pathogen in Leghorn chickens; Jackwood MW et al.; Experiments were conducted in order to produce bordetellosis in specific-pathogen-free (SPF) leghorn chickens . In the first and second of three experiments, young turkeys and chickens, respectively, were allotted into groups and challenged at 2 weeks of age with one of seven different isolates of Bordetella avium and two isolates of B . avium-like bacteria . Isolates of B . avium with the smooth colony type were pathogenic in turkeys but not in chickens . The B . avium-like bacteria and B . avium isolates with rough colonial morphology were nonpathogenic . In the third experiment, SPF leghorn chicks were vaccinated at 1 day of age with infectious bronchitis virus, Newcastle disease virus, or both and then challenged with B . avium at 7 days of age . Chickens vaccinated and then challenged with B . avium showed clinical signs and lesions similar to bordetellosis in turkeys . No clinical signs were observed in unchallenged chickens . Thus, we conclude that B . avium is an opportunistic pathogen in SPF leghorn chickens. Oral Microbiol Immunol, 1995 Apr, 10(2), 76 - 80 Demonstration of adherence properties of Porphyromonas gingivalis outer membrane vesicles using a new microassay; Duchesne P et al.; Vesicles made by Porphyromonas gingivalis possess several biological activities, including the ability to adhere to oral surfaces and to bacteria . In this study, a new and simple method was developed to measure the adherence capability of outer membrane vesicles from P . gingivalis . Vesicles were conjugated to fluorescent microspheres (0.7 micron) and added to wells of a Teflon-coated microscope slide previously covered with a variety of soluble ligands . After incubation and washes, the number of fluorescent microspheres per microscopic field were counted . Vesicle-coated microspheres attached best to gelatin (> 200 per field), whereas other compounds (such as fibronectin, fibrinogen, collagen and laminin) provided moderate attachment, and no attachment was observed to bovine serum albumin . Adherence to any of the tested ligands was not observed when fluorescent microspheres were conjugated to bovine serum albumin or lipopolysaccharides from P . gingivalis . The adherence of vesicle-coated microspheres to ligands was not significantly affected when the pH of the reaction mixture was between 4 and 10 . None of the tested carbohydrates lowered the attachment capability of vesicle-coated microspheres to substrates . When vesicle-coated microspheres were treated with trypsin and chymotrypsin or heated, this resulted in a significant loss of attachment, suggesting a possible involvement of proteinaceous molecules in the process . The present study confirms that vesicles of P . gingivalis are capable of attachment to various molecules and indicate their potential role in colonization. Resuscitation, 1995 Apr, 29(2), 157 - 68 The potential etiologic role of tumor necrosis factor in mediating multiple organ dysfunction in rats following intestinal ischemia-reperfusion injury; Yao YM et al.; Endogenous inflammatory cytokines may function as mediators in the development of remote organ damage in response to local ischemic insult . This study was designed to (a) explore the potential role of tumor necrosis factor (TNF) formation in the pathogenesis of systemic tissue injury, (b) determine the relationship between induction of TNF and gut-derived endotoxemia and/or bacterial translocation, and (c) evaluate the protective effect of anti-TNF monoclonal antibody (MoAb) for vital organs following intestinal ischemia-reperfusion in rats . Animals were subjected to superior mesenteric artery occlusion (SMAO) for 45 min . Systemic plasma TNF levels increased rapidly after the onset of reperfusion, reaching a peak value 2 h later (P < 0.01) . TNF elevation was found to be associated with gut origin endotoxemia, where the maximal TNF levels occurred approximately 2 h after the initial appearance of endotoxin in portal vein . Prophylactic treatment with anti-TNF MoAb markedly blunted the elevation in plasma TNF levels and afforded protection from the development of hypotension, vital organs dysfunction, and metabolic acidosis . Significant improvement in 48-h survival rate was observed by administration of anti-TNF MoAb prior to inducing ischemia (P = 0.007) . These findings suggest that intestinal ischemia-reperfusion could result in TNF production, which may play a key role in mediating subsequent septic response and systemic tissue injury . It seems likely that passage of endotoxin and bacteria from the gut can be responsible for the TNF formation Am J Infect Control, 1995 Apr, 23(2), 65 - 72 The effects of circuit and humidifier type on contamination potential during mechanical ventilation: a laboratory study; Gilmour IJ et al.; BACKGROUND: This study was undertaken because of concerns that ventilator humidifiers could be exacerbating the problem of nosocomial pneumonia in patients receiving mechanical ventilation . METHODS: Four different brands of humidifiers were used in conjunction with a siemens Servo 900B mechanical ventilator (Siemens Life Support Services, Solna, Sweden) . In the first part, the ventilator was operated with humidifiers filled with contaminated water at room temperature . The viability of airborne particles and the effect of flow rates on the number of particles produced were assessed . In the second part, we measured the effect of time and temperature on bacterial survival in humidifier chambers . Because only bubble-through humidifiers were determined to produce infectious particles, the speed with which a contaminated bubble-through humidifier could infect circuit condensate was also determined . Aliquots of chamber water and circuit condensate, as well as air samples and distal circuit swabs, were cultured . RESULTS: Humidifiers other than bubble-through humidifiers did not produce aerosols . Particle production by bubble-through humidifiers varied directly with flow rate (R2 = 0.91) . Chamber temperatures did not affect chamber colony counts except in bubble-through humidifiers . Although chamber colony counts in bubble-through humidifiers decreased with time, organisms remained viable throughout the study . When bubble-through humidifiers were heated, both condensate and effluent gas became heavily contaminated within minutes of flow initiation . CONCLUSIONS: Bubble-through humidifiers produce aerosols that readily contaminate both circuit condensate and effluent gas . Avoiding bubble-through humidifiers should improve patient safety while allowing changes in practice that can result in significant cost savings. Vaccine, 1995 Apr, 13(5), 455 - 62 Immune responses and protection against Bordetella pertussis infection after intranasal immunization of mice with filamentous haemagglutinin in solution or incorporated in biodegradable microparticles; Cahill ES et al.; The intranasal (i.n.) immunization of mice with Bordetella pertussis filamentous haemagglutinin (FHA) either as a solution or incorporated in biodegradable microparticles induced very similar immune responses . Both resulted in strong systemic IgG responses to FHA and good levels of anti-FHA IgG and IgA in the lungs of immunized mice . In comparison, the intraperitoneal (i.p.) immunization of mice with FHA, as a solution, engendered anti-FHA antibody responses which were stronger for serum IgG, similar for lung IgG and lower for lung IgA . The anti-FHA antibody levels, as measured by immunosorbent assay, were shown to correlate with their functional activity in the blocking of B . pertussis adhesion to HeLa tissue-culture cells . All three forms of immunization appeared to stimulate T-cell responses as assessed by in vitro antigen-specific spleen cell proliferation and IL-2 secretion indicative of a Th1 type response, however, cells from i.p . immunized mice only secreted low levels of IL-5 . All three methods of FHA immunization provided mice with significant protection against subsequent aerosol challenge with virulent B . pertussis . Mice which had been immunized intra-nasally eliminated the bacteria from their lungs slightly more rapidly than i.p . immunized mice, demonstrating the efficacy of intranasal administration of FHA in solution and in the more practical biodegradable microparticle form. Histochem J, 1995 Apr, 27(4), 253 - 71 The role of lectins in plant defence; Peumans WJ et al.; Recent progress in the search for the physiological role of plant lectins supports the idea that some of these proteins are involved in the defence mechanisms of the plant . To place the evidence in favour of such a defensive role in a broad perspective, a short overview is given of the most important plant pathogens and predators . In addition, the solutions that plants have developed to resist the continuous threat of a hostile environment are briefly discussed in relation to the protective role of proteins in general . The presumed involvement of plant lectins in defence mechanisms is first inferred from an analysis of the biochemical, physiological, cellular biological and molecular biological properties of plant lectins . Subsequently, the available experimental evidence for the involvement of lectins in the plant's defence against viruses, bacteria, fungi and herbivorous invertebrates and vertebrates is discussed in some detail . Since the defensive role of plant lectins is determined largely by their ability to recognize and bind foreign glycans, a brief discussion is given of how the basically protective properties of these proteins can be exploited for histochemical applications in biological and biomedical research. Ann Ital Med Int, 1995 Apr-Jun, 10(2), 113 - 8 {In vitro effects of peptides of the corticostatin/defensin family on production of mitogen-induced cytokines}; Masera RG et al.; Cytokines are autocrine, paracrine and endocrine glycoproteins that interact with specific cell receptors and have pleiotropic effects . Increasing evidence indicates that cytokines, immune interferon (IFN-gamma) and interleukin 6 (IL-6) among others, modulate hypothalamic-pituitary-adrenal function . Corticostatins/defensins are a family of cationic peptides recently isolated from phagocytic cells of myeloid lineage . Four peptides have been isolated from human neutrophils: HP-1, 2, 3 and 4 . As defensins they participate in immunosurveillance against viruses, bacteria and fungi . Some members of the family are also able to inhibit ACTH-induced steroidogenesis . Among human peptides, only HP-4 is corticostatic . We previously demonstrated that HP-1 and HP-4 inhibit in vitro the spontaneous and cytokine-inducible natural killer activity of human peripheral blood mononuclear cells (PBMC) and potentiate cortisol-dependent inhibition . The present work was carried out to determine whether two human corticostatins/defensins, HP-1 and HP-4, were able to modulate in vitro IFN-gamma and IL-6 production by human PBMC stimulated with phytohemagglutinin or Concanavalin A . IFN-gamma was titrated using biological assay with WISH cells as indicators and vesicular stomatitis virus as the challenge virus . IL-6 was measured by means of enzyme amplified sensitivity immunoassay . Both HP-1 and HP-4 significantly reduced cytokine production . Our data indicate that HP-1 and HP-4 are novel modulators of lymphocyte functions in vitro . Their depressing properties on ACTH-induced steroidogenesis and on cytokine production add complexity to neuroendocrine-immune circuits involving hypothalamic-pituitary-adrenal function. Semin Thorac Cardiovasc Surg, 1995 Apr, 7(2), 70 - 7 New approaches to prevention and treatment of nosocomial pneumonia; Bassin AS et al.; The critical care environment contributes to the development of respiratory infections . The mortality rate of nosocomial pneumonia is as high as 50% to 70%, and approximately half of the deaths are directly attributable to the presence of lung infection . Altered host defenses, emergence of bacterial resistance, and technological advances that result in various therapeutic interventions contribute to pneumonia and its associated mortality in the critically ill population . Nasogastric intubation, elevated gastric pH, large gastric volumes, and endotracheal intubation may all promote exposure of the lungs to large numbers of bacteria . An understanding of the pathogenesis of this serious illness could allow us to devise methods for curbing the incidence and severity of the disease . The clinical setting in which pneumonia develops can be a guide to the organisms responsible, and an understanding of this relationship is helpful in developing a treatment plan . The diagnosis of nosocomial pneumonia is not always accurate, and empiric therapy with appropriately selected antibiotics can be potentially life-saving. Boll Chim Farm, 1995 Apr, 134(4), 204 - 8 Synthetic approaches to some new sulfur-containing heterocycles of anticipated immunosuppressive activity; Girges MM et al.; Several new isolated or fused heterocyclic ring systems that accomodate the isothioureido functionality, often associated with immunosuppressive activity, were synthesized for possible use as immunosuppressive agents . Preparation of these anchored heterocycles was achieved via a multi-step synthesis starting with the key intermediate thiazolyl thiourea derivative (I) . Structure of the newly synthesized products was confirmed using both of elemental and spectral analyses. J Otolaryngol, 1995 Apr, 24(2), 105 - 10 Phagocytosis and oxidative burst of granulocytes in the upper respiratory tract in chronic and acute inflammation; Muns G et al.; Acute diseases and chronic conditions may be associated with altered neutrophil functions . The aim of this investigation was to study the number and function of granulocytes in the upper respiratory tract in acute and chronic airway inflammation . The 136 participating subjects were assigned to 6 groups; group 1: controls; group 2: smokers, < 10 cigarettes/day; group 3: smokers, > 10 cigarettes/day; group 4: patients with allergic rhinitis; group 5: patients with obstructive sleep apnea; group 6: patients with acute viral upper respiratory tract infection . Whole blood and nasal lavage fluid were obtained and a cell count was performed . Phagocytosis of opsonized bacteria and the oxidative burst was measured . Total cell counts and absolute number of neutrophils in nasal lavage (NAL), but not in blood, were elevated in all groups compared to controls . Highest increases of neutrophils were observed in group 4 (7-fold) and group 6 (25-fold), p < .01 . Phagocytosis and oxidative burst of granulocytes recovered by NAL were found to be increased in group 4 (25% and 15%, respectively) and group 6 (35% and 21%) only, but decreased in the other groups . We conclude that conditions leading to acute or chronic inflammation in the upper respiratory tract are associated with altered functions of granulocytes recovered by NAL but not blood, suggesting local modulating factors . We speculate that chronic inflammation, in contrast to acute conditions, is associated with impaired granulocyte function. Clin Podiatr Med Surg, 1995 Apr, 12(2), 183 - 7 Paronychia; Black JR; Paronychia is defined most often in terms of inflammation and is classified as congenital, acute-acquired, or chronic-acquired . The cause for acquired paronychia usually is trauma, and middle-aged women are at greatest risk . Chronic paronychia has been associated with numerous occupations where fluids are the common denominator . Mixed infections usually are found in chronic paronychia, including bacteria, fungus, and yeast . Depending upon the type of paronychia and associated organisms, therapy includes footgear considerations, pharmacology measures, and surgical measures. J Bioenerg Biomembr, 1995 Apr, 27(2), 207 - 14 Uroporphyrinogen decarboxylase; Elder GH et al.; Uroporphyrinogen decarboxylase (EC 4.1.1.37) catalyzes the decarboxylation of uroporphyrinogen III to coproporphyrinogen III . The amino acid sequences, kinetic properties, and physicochemical characteristics of enzymes from different sources (mammals, yeast, bacteria) are similar, but little is known about the structure/function relationships of uroporphyrinogen decarboxylases . Halogenated and other aromatic hydrocarbons cause hepatic uroporphyria by decreasing hepatic uroporphyrinogen decarboxylase activity . Two related human porphyrias, porphyria cutanea tarda and hepatoerythropoietic porphyria, also result from deficiency of this enzyme . The roles of inherited and acquired factors, including iron, in the pathogenesis of human and experimental uroporphyrias are reviewed. J Bioenerg Biomembr, 1995 Apr, 27(2), 151 - 9 5-Aminolevulinate synthase and the first step of heme biosynthesis; Ferreira GC et al.; 5-Aminolevulinate synthase catalyzes the condensation of glycine and succinyl-CoA to yield 5-aminolevulinate . In animals, fungi, and some bacteria, 5-aminolevulinate synthase is the first enzyme of the heme biosynthetic pathway . Mutations on the human erythroid 5-aminolevulinate synthase, which is localized on the X-chromosome, have been associated with X-linked sideroblastic anemia . Recent biochemical and molecular biological developments provide important insights into the structure and function of this enzyme . In animals, two aminolevulinate synthase genes, one housekeeping and one erythroid-specific, have been identified . In addition, the isolation of 5-aminolevulinate synthase genomic and cDNA clones have permitted the development of expression systems, which have tremendously increased the yields of purified enzyme, facilitating structural and functional studies . A lysine residue has been identified as the residue involved in the Schiff base linkage of the pyridoxal 5'-phosphate cofactor, and the catalytic domain has been assigned to the C-terminus of the enzyme . A conserved glycine-rich motif, common to all aminolevulinate synthases, has been proposed to be at the pyridoxal 5'-phosphate-binding site . A heme-regulatory motif, present in the presequences of 5-aminolevulinate synthase precursors, has been shown to mediate the inhibition of the mitochondrial import of the precursor proteins in the presence of heme . Finally, the regulatory mechanisms, exerted by an iron-responsive element binding protein, during the translation of erythroid 5-aminolevulinate synthase mRNA, are discussed in relation to heme biosynthesis. J Comp Pathol, 1995 Apr, 112(3), 307 - 18 Helicobacter-like organisms: histopathological examination of gastric biopsies from dogs and cats; Hermanns W et al.; Gastric biopsies from the fundic gland region of 122 dogs and 127 cats were subjected to histopathological examination . The aim of the study was to determine infection rates and degrees of colonization by Helicobacter-like organisms (HLOs), and to ascertain their possible relationship to histopathological changes . In all, 82% of the dogs and 76% of the cats had an HLO infection . The most striking histopathological changes were glandular degeneration with accumulation of lymphocytes and neutrophilic granulocytes (dogs, 21%; cats, 39%), fibrosis of the lamina propria mucosae (dogs, 41%; cats, 58%), oedema in the lamina propria mucosae (dogs, 54%; cats, 23%), lymphoid follicles (dogs, 17%; cats, 19%) and lymphoplasmacytic infiltrates . A relation between the degree of colonization by HLOs and the extent of histopathological changes could only be discovered in the cats . It was not possible to ascertain whether these bacteria, irrespective of the degree of colonization, were responsible for the histopathological changes in the dogs. Int Dent J, 1995 Apr, 45(2), 135 - 40 Some physical and biological properties of glass ionomer cement; Mount GJ; It has become apparent through both clinical use and laboratory experiments that the glass ionomer cements have several highly desirable properties . They show a continuing fluoride release and the ability to take up further fluoride under favourable conditions . The presence of fluoride also helps to inhibit plaque formation . The adhesion between tooth structure and cement also results in almost complete prevention of the bacterial micro-leakage . Also, the cement itself is so highly bio-compatible that it is now being used as a bone substitute and it has become apparent that there is no need to place a sub-lining under a glass ionomer restoration . Recent research is leading to the development of self curing cements with enhanced physical properties so that, in the presence of the above advantages, their use in clinical dentistry is rapidly expanding . Glass ionomer cements are of great value for any restoration which is not under undue occlusal stress and they work well also as a long term temporary restoration in the presence of a high caries rate, where zinc oxide and eugenol used to be the material of choice. Dtsch Tierarztl Wochenschr, 1995 Apr, 102(4), 163 - 4 Effect of n-butyric acid on epithelial cell proliferation of pig colonic mucosa in short-term culture; Sakata T et al.; Short-chain fatty acids (SCFA) such as acetic, propionic and n-butyric acids produced by hindgut bacteria stimulate gut epithelial cell proliferation through afferent neural and efferent non-neural systemic transmissions beside a probable local mechanism . In the present study, we developed an experimental system using pig hindgut mucosa in short term culture to clarify the mechanism of the local trophic effect of SCFA . Pig mucosal tissue pieces of the distal colon were cultured in RPMI 1640 medium containing glutamine, 20% (v/v) newborn calf serum and n-butyric acid (0, 0.5, 1.0 or 5.0 mmol/L) . Crypt cell production rate from 0.5 to 3.5 and from 21 to 24 hours of culture was measured . Butyric acid increased crypt cell production rate of pit distal colon only at 5 mmol/L . The effect of butyric acid did not differ between samples of different length of n-butyric acid exposure . The effect of n-butyric acid in this study resembled to that found in human biopsied specimens of the colon . The present results also indicated that epithelial cells of pig colonic mucosa in short-term culture presented here retained the proliferative activity and the responsiveness to n-butyric acid. Dtsch Tierarztl Wochenschr, 1995 Apr, 102(4), 152 - 4 Intestinal methanogenesis in primates--a genetic and evolutionary approach; Hackstein JH et al.; The presence of significant numbers of intestinal methanogens among vertebrates does neither depend on elaborated morphological structures nor on predominantly plant-based diets . Phylogenetic rather than dietary restrictions limit the occurrence of methanogenic bacteria also in the hindguts of primates . The Old-World monkeys are methanogenic--with the only exceptions of Cheirogaleid lemurs and bush-babies . In contrast, among New World monkeys the lack of intestinal methanogens is observed frequently in capuchins and marmosets . Since the absence of methanogens does not parallel distinct morphological, physiological, or ethological characteristics, it is likely that methanogenesis depends on a hereditary predisposition . In humans, methane-producers account for approximately 50% of the European populations . In this study, 56 individuals belonging to 5 families were studied for the occurrence of methane in the breath . The results of this screen are compatible with the assumption that the trait "methane emission" segregates as an autosomal, dominant character . Our findings suggest a high specificity of the symbiosis between primates and methanogenic bacteria . Therefore, the persistence of significant numbers of methanogens in the hindgut might be facilitated by a specific receptor for methanogenic bacteria. J Clin Microbiol, 1995 Apr, 33(4), 893 - 7 Detection of Mycoplasma hyopneumoniae in nose swabs from pigs by in vitro amplification of the 16S rRNA gene; Mattsson JG et al.; We report the use of PCR to detect DNA from Mycoplasma hyopneumoniae, the etiological agent of enzootic porcine pneumonia . A primer set was designed for the amplification of a 649-bp fragment of the 16S rRNA gene from M hyopneumoniae . The PCR product was identified by ethidium bromide staining after gel electrophoresis and by Southern hybridization with an M . hyopneumoniae-specific oligonucleotide probe . No amplification was observed from any other porcine bacteria tested, including several closely related mycoplasmas . It was also possible to demonstrate the presence of M . hyopneumoniae in bronchial lavage samples and lung tissue samples from experimentally infected pigs . Furthermore, the PCR system was used for analysis of nasal samples obtained from pigs in a fattening herd . By this method, we were able to detect M . hyopneumoniae in nose swabs from naturally infected pigs . However, our results suggest that M . hyopneumoniae can be detected in the nasal cavities only during a limited time period. Aust Dent J, 1995 Apr, 40(2), 129 - 34 Cellular adhesion molecules on periodontal lymphocytes; Gemmell E et al.; T cell induced differentiation of B cells has been shown to be dependent on the CD2/LFA-3 and LFA-1/ICAM-1 pathways . Flow cytometric analysis was used to examine these adhesion molecules on T and B cells extracted from gingival tissues before and after stimulation with the putative periodontopathic bacteria, Porphyromonas gingivalis and Fusobacterium nucleatum . Adhesion molecule expression on peripheral blood cells from healthy adults was used as a control . Approximately 50 per cent of B cells extracted from gingival tissues expressed LFA-3 and ICAM-1 compared with 30 per cent positive peripheral blood B cells . Around 50 per cent of gingival T cells expressed CD2 relative to 76 per cent positive peripheral blood T cells . However, 40-50 per cent of both gingival and peripheral blood T cells expressed LFA-1 . There was no difference in the expression of adhesion molecules on T and B cells extracted from health/marginal gingivitis or adult periodontitis lesions . After stimulation of gingival cells in vitro, the per cent CD2 positive T cells and LFA-3 and ICAM-1 positive B cells remained relatively stable over the six-day culture period, although P . gingivalis and F . nucleatum appeared to induce an increase in the percentage of gingival T cells expressing LFA-1 . In contrast to the gingival lymphocytes, stimulation of peripheral blood cells resulted in an increase in the per cent CD2 positive T cells, LFA-3 and ICAM-1 positive B cells, with a decrease in LFA-1 positive T cells . The results therefore demonstrated that gingival T and B cells express adhesion molecules in vivo.(ABSTRACT TRUNCATED AT 250 WORDS) J Infect Dis, 1995 Apr, 171(4), 916 - 23 Etiology of cat scratch disease: comparison of polymerase chain reaction detection of Bartonella (formerly Rochalimaea) and Afipia felis DNA with serology and skin tests; Bergmans AM et al.; To determine the role of Bartonella (formerly Rochalimaea) species and Afipia felis in cat scratch disease (CSD), two polymerase chain reaction (PCR) hybridization assays were developed to detect DNA from these organisms . These assays were applied on 89 pus aspirates from skin test-positive CSD patients (group 1) and on 137 pus and lymph node specimens from CSD suspects (group 2) . Bartonella DNA was detected in 96% of the samples from group 1 patients and in 60% of group 2 samples; however, A . felis DNA could not be detected in any clinical samples . These results suggest that CSD is caused by bartonellae and that A . felis does not play a significant role in this zoonosis . A strong correlation between Bartonella PCR positivity and Bartonella henselae antibody titer was found . Comparison of CSD skin test results with those obtained by Bartonella PCR suggests a low sensitivity of the skin test. J Biol Chem, 1995 Mar 31, 270(13), 7227 - 32 Isoform-specific differences in the potencies of murine protein kinase inhibitors are due to nonconserved amino-terminal residues; Gamm DM et al.; We provide here a detailed characterization of two isoforms of the protein kinase inhibitor (PKI) protein of cAMP-dependent protein kinase that have dramatically different inhibition constants . Murine PKI beta 1 possesses a 32-fold higher Ki than murine PKI alpha as determined by Henderson analysis . This finding led to the investigation of C subunit.PKI interactions involving nonconserved regions in the carboxyl and amino termini of murine PKI alpha and PKI beta 1 . Chimeric cDNAs coding for amino acid sequences from both PKI isoforms were constructed and expressed in bacteria . Surprisingly, exchanging the carboxyl-terminal two-thirds of PKI alpha and PKI beta 1 has relatively little effect on the inhibition constants of the two isoforms . Similarly, introducing amino acid residues corresponding to a beta-turn region of PKI alpha into PKI beta 1 fails to lower PKI beta 1 inhibition constants . However, introducing the amino-terminal alpha-helical region of PKI alpha into PKI beta 1 reduces the Ki and IC50 of PKI beta 1 to values identical with full length PKI alpha . Site-directed mutagenesis of specific residues within this region implicates the presence of a tyrosine at position 7 in PKI alpha as a major contributor to its enhanced inhibitory potency . The results of this study suggest that variations in C subunit.PKI interactions within an amino-terminal alpha-helix provide a major mechanism for altering the inhibitory properties of PKI isoforms. J Biol Chem, 1995 Mar 31, 270(13), 7156 - 60 A dopamine-responsive domain in the N-terminal sequence of Pit-1 . Transcriptional inhibition in endocrine cell types; Lew AM et al.; The POU transcription factor Pit-1 activates the prolactin gene in pituitary lactotrophs and may integrate responses of the gene to external signals . To study the role of Pit-1 in dopaminergic inhibition of the prolactin gene, we transiently transfected Pit-1 and dopamine D2 receptor vectors into a series of heterologous cell lines and examined dopamine regulation of the prolactin gene promoter . Regulation was Pit-1-dependent in all cell lines tested . Moreover, dopamine responsiveness was cell type-specific: stimulatory in fibroblasts (COS-7) and muscle-type cells (P19/Me2SO-induced) and inhibitory in pancreatic endocrine (RIN, InR1-G9) and neural-like (P19/retinoic acid-induced) cells . Because dopaminergic responses in Pit-1-transfected RIN cells paralleled those in pituitary GH4 cells, the islet cell line was used to test for sequences in Pit-1 that mediate negative hormone signals . Dopamine responsiveness of the Pit-1 transactivation domain (residues 8-80) was examined using a chimeric LexA construct . LxPit-1, LxSp1, and Lx-glucocorticoid receptor fusions all activated basal transcription, but only LxPit-1 was regulated by dopamine . Regulatory responses of LxPit-1 and full-length Pit-1 were quantitatively similar . In addition, gain-of-function G alpha mutants that inhibit Pit-1-dependent promoters in GH4 cells also suppressed selectively Pit-1- or LxPit-1-dependent promoters in RIN cells . This demonstrates that Pit-1 can function as a specific target for distinct inhibitory G protein signals . Interestingly, Pit-1 sequences N-terminal to the DNA-binding POU domain appear to be sufficient in mediating regulation by these pathways. J Biol Chem, 1995 Mar 31, 270(13), 7055 - 60 Purification and photoaffinity labeling of herpes simplex virus type-1 thymidine kinase; Rechtin TM et al.; The molecular basis for the treatment of human herpesviruses with nucleoside drugs is the phosphorylation of these drugs by the viral-encoded thymidine kinases . In order to better understand the structural and enzymatic mechanisms by which herpesviral thymidine kinases recognize their substrates, photoaffinity labeling with {alpha-32P}5-azido-2'-deoxyuridine-5'-monophosphate and { gamma-32P}8-azidoadenosine-5'-triphosphate was used to characterize the thymidine, thymidylate, and ATP active sites of the herpes simplex virus-1 (HSV-1) thymidine kinase . For this study, HSV-1 thymidine kinase and a site-specific mutant enzyme (C336Y, known to confer acyclovir resistance) were expressed in bacteria and purified by a rapid, two-step protocol . The specificity of photoaffinity labeling of these HSV-1 thymidine kinases was demonstrated by the ability of site-directed substrates such as thymidine, thymidylate, acyclovir, 5-bromovinyl-2'-deoxyuridine, and ATP to inhibit photoinsertion . Differences in inhibition patterns of photoaffinity labeling correlated with kinetic differences between the wild-type and C336Y HSV-1 thymidine kinases . Cumulative results suggest that the acyclovir-resistant cysteine 336 mutation primarily affects the ATP binding site; yet it also leads to alteration in the binding affinity of nucleoside drugs in the thymidine site . In this study, azidonucleotide photoaffinity analogs are shown to be effective tools for studying the active-site environment of HSV-1 thymidine kinase and related site-specific mutants. Proc Natl Acad Sci U S A, 1995 Mar 28, 92(7), 2647 - 51 Chloroplast gene sequence data suggest a single origin of the predisposition for symbiotic nitrogen fixation in angiosperms; Soltis DE et al.; Of the approximately 380 families of angiosperms, representatives of only 10 are known to form symbiotic associations with nitrogen-fixing bacteria in root nodules . The morphologically based classification schemes proposed by taxonomists suggest that many of these 10 families of plants are only distantly related, engendering the hypothesis that the capacity to fix nitrogen evolved independently several, if not many, times . This has in turn influenced attitudes toward the likelihood of transferring genes responsible for symbiotic nitrogen fixation to crop species lacking this ability . Phylogenetic analysis of DNA sequences for the chloroplast gene rbcL indicates, however, that representatives of all 10 families with nitrogen-fixing symbioses occur together, with several families lacking this association, in a single clade . This study therefore indicates that only one lineage of closely related taxa achieved the underlying genetic architecture necessary for symbiotic nitrogen fixation in root nodules. Orv Hetil, 1995 Mar 26, 136(13), 659 - 62 {Detection of Chlamydia trachomatis in chronic prostatitis by in situ hybridization (preliminary methodical report)}; Kadar A et al.; (Preliminary methodical report) . In situ hybridization of Chlamydia trachomatis in formalin fixed, paraffin embedded specimens from the Urology Clinic were performed in 2nd Department of Pathology Semmelweis University of Medicine by a biotin labelled DNA probe . Chlamydia trachomatis is suspected to be responsible for the chronic abacterial inflammation of the prostate besides Ureaplasma urealyticum and Mycoplasmae . According to our retrospective study out of 79 biopsy specimens 34 had the diagnosis of chronic abacterial prostatitis . We examined 11 specimens of them . Bacteria were not identified . Three specimens were positive for Chlamydia trachomatis which were improved by transmission electron microscope . The age of patients was between 59 and 81 years . The detection of chlamydia infection rises the suspicion of a "healthy" career and especially at younger age could cause Chlamydia trachomatis associated genital disease . The undiagnosed and untreated diseases may lead to infertility. J Biol Chem, 1995 Mar 24, 270(12), 6456 - 63 Human intestinal H+/peptide cotransporter . Cloning, functional expression, and chromosomal localization; Liang R et al.; In mammalian small intestine, a H(+)-coupled peptide transporter is responsible for the absorption of small peptides arising from digestion of dietary proteins . Recently a cDNA clone encoding a H+/peptide cotransporter has been isolated from a rabbit intestinal cDNA library (Fei, Y.J., Kanai, Y., Nussberger, S., Ganapathy, V., Leibach, F.H., Romero, M.F., Singh, S.K., Boron, W . F., and Hediger, M . A . (1994) Nature 368, 563-566) . Screening of a human intestinal cDNA library with a probe derived from the rabbit H+/peptide cotransporter cDNA resulted in the identification of a cDNA which when expressed in HeLa cells or in Xenopus laevis oocytes induced H(+)-dependent peptide transport activity . The predicted protein consists of 708 amino acids with 12 membrane-spanning domains and two putative sites for protein kinase C-dependent phosphorylation . The cDNA-induced transport process accepts dipeptides, tripeptides, and amino beta-lactam antibiotics but not free amino acids as substrates . The human H+/peptide cotransporter exhibits a high degree of homology (81% identity and 92% similarity) to the rabbit H+/peptide cotransporter . But surprisingly these transporters show only a weak homology to the H(+)-coupled peptide transport proteins present in bacteria and yeast . Chromosomal assignment studies with somatic cell hybrid analysis and in situ hybridization have located the gene encoding the cloned human H+/peptide cotransporter to chromosome 13 q33-->q34. Nature, 1995 Mar 23, 374(6520), 371 - 5 DNA-bend modulation in a repressor-to-activator switching mechanism; Ansari AZ et al.; Recent discoveries of activator proteins that distort DNA but bear no obvious activation domains have focused attention on the role of DNA structure in transcriptional regulation . Here we describe how the transcription factor MerR can mediate repression as well as activation through stereospecific modulation of DNA structure . The repressor form of MerR binds between the -10 and -35 promoter elements of the bacterial mercury-detoxification genes, PT, allowing RNA polymerase to form an inactive complex with PT and MerR at this stress-inducible promoter . Upon mercuric ion binding, Hg-MerR converts this polymerase complex into the transcriptionally active or 'open' form . We show here that MerR bends DNA towards itself in a manner similar to the bacterial catabolite-activator protein CAP, namely at two loci demarked by DNase I sensitivity, and that the activator conformation, Hg-MeR, relaxes these bends . This activator-induced unbending, when coupled with the previously described untwisting of the operator, remodels the promoter and makes it a better template for the poised polymerase. Science, 1995 Mar 17, 267(5204), 1655 - 8 Development of a mouse model of Helicobacter pylori infection that mimics human disease; Marchetti M et al.; The human pathogen Helicobacter pylori is associated with gastritis, peptic ulcer disease, and gastric cancer . The pathogenesis of H . pylori infection in vivo was studied by adapting fresh clinical isolates of bacteria to colonize the stomachs of mice . A gastric pathology resembling human disease was observed in infections with cytotoxin-producing strains but not with noncytotoxic strains . Oral immunization with purified H . pylori antigens protected mice from bacterial infection . This mouse model will allow the development of therapeutic agents and vaccines against H . pylori infection in humans. Oncogene, 1995 Mar 16, 10(6), 1167 - 74 The DNA binding activity of wild type p53 is modulated by blocking its various antigenic epitopes; Wolkowicz R et al.; Interaction of wild type p53 with specific DNA target sequences, which is dictated by several structural domains, can be modified by blocking the different antigenic epitopes of the protein . Comparison of p53 protein expressed by recombinant bacteria (wtp53-Bac) to that produced in an eukaryotic system by a vaccinia expression vector (wtp53-Vac), indicated that only the later exhibited spontaneous DNA-binding activity . Furthermore, DNA-binding patterns of these wild type p53 proteins were affected differently by their interactions with monoclonal anti-p53 antibodies recognizing individual antigenic epitopes of the molecule . While the vaccinia derived p53 that spontaneously bound DNA is supershifted by the N'-terminal specific antibodies PAb-248, the bacterial derived p53 protein that retains this antigenic epitope but does not bind DNA spontaneously, is not affected . The C'-terminal specific PAb-421 antibodies accelerated binding of the bacterial p53 protein and modified the pattern of the interaction of the vaccinia derived p53 DNA . DNA-binding patterns generated by PAb-421 and PAb-248, suggest that either interaction of wild type p53 is dependent on modification of the p53 protein or that it interacts with cellular factors which their activity can be mimicked by PAb-421 . Saturation of both types of wild type p53 with several anti-p53 monoclonal antibodies directed against the wild type p53 specific epitope that maps to the N'-terminal border of the DNA-binding region, blocked specific DNA-binding . The fact that most p53 mutants have lost the wild type p53 conformation specific epitope coupled with the observation that blocking of this site by binding specific antibodies, prevents the interaction of wild type p53 with DNA, suggests that maintaining the correct structural conformation of this site is central for DNA-binding activity . The wild type specific epitope which maps to the N'-terminal border of the DNA-binding region is neighboring the first beta-strand detected by the recent crystallographic analysis. Rev Prat, 1995 Mar 15, 45(6), 729 - 32 {Mycobacterial infections in AIDS}; Perronne C; Mycobacterial infections are the main cause of bacterial infections during HIV infection . The HIV infection epidemic contributed to the worldwide resurgence of the tuberculous endemic . The clinical presentation of tuberculosis is often atypical and extrapulmonary . The general guidelines for treatment are not modified, but HIV infection often worsens the tolerance of treatment . A poor compliance to treatment is the main cause of relapse or of selection of resistant mutants . The two main points for the control of tuberculosis in the era of HIV are a free access of patients to antituberculosis drugs, and a surveillance of the compliance until treatment is completed . Atypical mycobacteria, mainly represented by Mycobacterium avium, are bacteria from the environment, poorly virulent, responsible for infections occurring late in the course of AIDS . Current treatments use combinations including clarithromycin. Eur J Biochem, 1995 Mar 15, 228(3), 971 - 5 Processing and hydrolytic mechanism of the cgkA-encoded kappa-carrageenase of Alteromonas carrageenovora; Potin P et al.; The cgkA gene of Alteromonas carrageenovora encodes a kappa-carrageenase with a predicted mass of 44212 Da, much larger than the 35 kDa estimated from SDS/PAGE of the protein purified from culture supernatants . Immunoblotting experiments showed the presence of a protein of 44 +/- 2 kDa in both native and recombinant bacterial intracellular extracts, suggesting that the kappa-carrageenase is produced as a preproprotein which undergoes proteolytic processing twice during secretion . To determine the exact site of C-terminal cleavage, the precise mass of the purified extracellular kappa-carrageenase was measured by electrospray-ionization/mass spectrometry and found to be 31,741 +/- 3 Da . The mature kappa-carrageenase of A . carrageenovora thus appears to be composed of 275 amino acids, from residue Ala26 to residue Asn301 of the cgkA gene product . To assess the molecular mechanism of this member of family 16 of glycosyl hydrolases, hydrolysis of neocarrahexaitol by the kappa-carrageenase was monitored by gel filtration chromatography and 13C-NMR . Results show that neocarrabiitol and beta-neocarratetraose are initially formed, demonstrating that the enzyme operates with a molecular mechanism retaining the anomeric configuration . Consistent with this result, the enzyme was also shown to be able to catalyze transglycosylation. J Biol Chem, 1995 Mar 3, 270(9), 4805 - 12 Characterization of Rad, a new member of Ras/GTPase superfamily, and its regulation by a unique GTPase-activating protein (GAP)-like activity; Zhu J et al.; We have recently identified a new member of the Ras/GTPase superfamily termed Rad which has unique sequence features and is overexpressed in the skeletal muscle of humans with type II diabetes (Reynet, C., and Kahn, C . R . (1993) Science, 262, 1441-1444) . When expressed in bacteria as a glutathione S-transferase fusion protein, Rad bound {alpha-32P}GTP quickly and saturably . Binding was specific for guanine nucleotides and displayed unique magnesium dependence such that both GTP and GDP binding were optimal at relatively high Mg2+ concentrations (1-10 mM) . Rad had low intrinsic GTPase activity which was greatly enhanced by a GTPase-activating protein (GAP) activity present in various tissues and cell lines . Several known GAPs had no stimulatory effect toward Rad . Conversion of Ser to Asn at position 66 in Rad (equivalent to position 12 in Ras) resulted in a total loss of GTP binding . Mutation of Pro61 (equivalent to Gly12 in Ras) or Gln109 (equivalent to Gln61 in Ras) had no effect on Rad GTPase activity, whereas creation of a double mutation at these positions resulted in exceptionally high intrinsic GTPase activity . In vitro, Rad was phosphorylated by the catalytic subunit of cAMP-dependent protein kinase (PK) . Phosphopeptide mapping indicated two PKA phosphorylation sites near the COOH terminus . Rad also co-precipitated a serine/threonine kinase activity from extracts of various tissues and cell lines which catalyzed phosphorylation on Rad but was not inhibited by PKA inhibitor . Thus, Rad is a GTP-binding protein and a GTPase which has some structure/function similarities to Ras, but displays unique features . Rad may also be phosphorylated on serine/threonine residues by PKA and other kinases, as well as regulated by its own GAP which is present in many tissues and cell types. J Biol Chem, 1995 Mar 3, 270(9), 4457 - 65 Sequence identity and antigenic cross-reactivity of white face hornet venom allergen, also a hyaluronidase, with other proteins; Lu G et al.; White face hornet (Dolichovespula maculata) venom has three known protein allergens which induce IgE response in susceptible people . They are antigen 5, phospholipase A1, and hyaluronidase, also known as Dol m 5, 1, and 2, respectively . We have cloned Dol m 2, a protein of 331 residues . When expressed in bacteria, a mixture of recombinant Dol m 2 and its fragments was obtained . The fragments were apparently generated by proteolysis of a Met-Met bond at residue 122, as they were not observed for a Dol m 2 mutant with a Leu-Met bond . Dol m 2 has 56% sequence identity with the honey bee venom allergen hyaluronidase and 27% identity with PH-20, a human sperm protein with hyaluronidase activity . A common feature of hornet venom allergens is their sequence identity with other proteins in our environment . We showed previously the sequence identity of Dol m 5 with a plant protein and a mammalian testis protein and of Dol m 1 with mammalian lipases . In BALB/c mice, Dol m 2 and bee hyaluronidase showed cross-reactivity at both antibody and T cell levels . These findings are relevant to some patients' multiple sensitivity to hornet and bee stings. Commun Dis Rep CDR Rev, 1995 Mar 3, 5(3), R40 - 4 How many people in England and Wales risk infection from injecting drug use? Durante AJ, Heptonstall J. Injecting drug users who share equipment may transmit and acquire bloodborne virus infections, including HIV, hepatitis B virus, and hepatitis C virus . Even without sharing, injection with non-sterile equipment, drugs, or mixing agents may result in infection due to bacteria or fungi . Estimates of the number of people who are currently at risk of infection from injecting drug use are needed in order to plan services and care, and to interpret surveillance data . This paper examines the data from registries of drug use and two recent surveys of the general population from which estimates of the number of current injecting drug users in England and Wales have been derived . Drug registries include only those whose drug use is identified during contact with drug or medical services, so these sources provide minimum estimates but may be used to monitor trends: 25,706 drug users in England and Wales were notified to the Home Office in 1993, 12,253 of whom were current injectors . Estimates derived from surveys of the general population suggest, however, that between 51,900 (95% confidence interval (CI): 33,000-71,600) and 77,700 (95% CI: 4100-151,200) people in England and Wales are at risk of infection from current injecting drug use, of whom between 10,400 (95% CI: 7200-13,800) and 15,500 (95% CI: 800-30,200) are at risk of bloodborne virus infections as a result of sharing injecting equipment . In the 16 to 34 year age group about one in 200 men, and one in 400 to 500 women may be current injectors.(ABSTRACT TRUNCATED AT 250 WORDS) Scanning Microsc, 1995 Mar, 9(1), 199 - 205; discussion 205-6 High resolution electron microscopy of enamel crystallites demineralized by initial dental caries; Hayashi Y; Acids produced by various oral bacteria cause mineral loss and crystallite dissolution during the development of enamel caries . In order to demonstrate this phenomenon, the initial disappearance of lattice fringes and the formation of a central perforation in crystallites were examined by high resolution electron microscopy (HREM) in initial enamel caries without macroscopic tissue evidence of destruction . Ultrathin sections were also examined by selected area electron diffraction to reveal the mineral phase of the surface layer in carious enamel . A marked variation in the dissolution pattern was disclosed in the initial carious lesions . HREM revealed that disappearance of the lattice fringes from the lateral portion of the crystallites was predominant in the superficial layer covering the lesion, while central perforation of crystallites mainly occurred in the subsurface prismatic region . The beginning of the central dissolution occurred at the dislocated area where lattice striations appeared to be disordered . Selected area electron diffraction of the gradually demineralized enamel revealed a pattern consistent with hydroxyapatite (OH-AP) or fluorapatite (F-AP) mineral . These findings suggest that the susceptibility to caries of enamel crystallites is spatially and temporally different during the progression of the caries . Furthermore, the formation of central perforations and the consequent easy intracrystalline diffusion of acids might induce rapid crystallite dissolution. Riv Eur Sci Med Farmacol, 1995 Mar-Jun, 17(2-3), 85 - 9 A retrospective study on the incidence of lymphocytic gastritis in patients with Helicobacter pylori infection; Sirigu F et al.; To investigate the relationship between lymphocytic gastritis and H . pylori infection, antral biopsy specimens from 162 patients with histological evidence of H . pylori infection, and from 177 subjects without colonization were retrospectively examined . Among patients with antral colonization, lymphocytic gastritis was identified in 8 cases (4.9%) . Lymphocytic gastritis was not detected in specimens without H . pylori infection . Quantitation of the epithelial lymphocytic infiltration was carried out in all specimens and showed that cases with lymphocytic gastritis had a mean of 35.3 lymphocytes per 100 epithelial cells, compared with a mean of 2.6 and 2.4 respectively in chronic gastritis and normal mucosa . Among the 331 patients without lymphocytic gastritis, specimens colonized by H . pylori had a mean of epithelial lymphocyte counts significantly higher than those observed in both chronic gastritis and normal mucosa . These findings show a positive correlation between antral colonization by H . pylori and occurrence of lymphocytic gastritis, and suggest that the bacteria may have an important role in the induction of such particular form of antral inflammation. J Trauma, 1995 Mar, 38(3), 344 - 9 Prospective comparison of two management strategies of central venous catheters in burn patients; Kealey GP et al.; OBJECTIVE: Central venous catheters (CVCs) are associated with sepsis in burn patients . This study was undertaken to compare two strategies of CVC management in patients with major burn injuries . DESIGN: Forty-two burn patients with major burn injuries were randomly assigned to undergo site change every 48 hours of the CVC or to undergo wire guide exchange of the CVC every 48 hours at the same site . MATERIALS AND METHODS: Catheter insertion site, distance from the burn wound, cultures of catheter tips, and blood cultures were obtained from all patients in a prospective manner . MEASUREMENTS AND MAIN RESULTS: There was no difference in the incidence of CVC sepsis between the two groups studied . CVCs inserted less than 5 cm from the burn wound developed bacterial contamination at an earlier time than CVCs inserted more than 5 cm from the burn wound . CONCLUSIONS: There was no advantage to changing the CVC insertion site every 48 hours . Changing the CVC using the wire guide technique did not prevent, nor predict, CVC bacterial contamination. FASEB J, 1995 Mar, 9(5), 371 - 8 Role of chaperones in the biogenesis and maintenance of the mitochondrion; Martinus RD et al.; All cells depend on correctly folded proteins for optimal function . A central question in cellular biology is how such folded structures are formed and maintained, a process that is now recognized to rely heavily on a group of proteins called molecular chaperones . Molecular chaperones constitute distinct families of proteins that are ubiquitous and highly conserved from bacteria to humans . They appear to bind nonnative conformations of most, if not all, proteins, thereby preventing their aggregation and subsequent inactivation . The chaperones not only protect newly synthesized proteins during transport and folding, but also serve to maintain the cell in a healthy state during exposure to a multitude of stress conditions . Accordingly, chaperones are expressed constitutively, but their synthesis is further enhanced during stress conditions . Detailed insights into the role of molecular chaperones have come from studies of mitochondrial protein biogenesis, a process in which chaperones act as unfoldases, pulling devices, and foldases . In this review we summarize these developments and further discuss the potential role of chaperones in mitochondrial DNA metabolism and human mitochondrial disease states. Medicine (Baltimore), 1995 Mar, 74(2), 83 - 8 Infections with Chryseomonas luteola (CDC group Ve-1) and flavimonas oryzihabitans (CDC group Ve-2); Rahav G et al.; We have described 15 cases of infection due to Chryseomonas luteola and Flavimonas oryzihabitans isolated between May 1990 and May 1994 . These infections were often associated with the presence of a foreign body, especially central venous access and joint prosthesis . The high frequency of isolating C . luteola and F . oryzihabitans probably expresses the awareness of the clinicians and the laboratory to the importance of these bacteria, especially in patients with the presence of a foreign material. J Nutr, 1995 Mar, 125(3 Suppl), 757S - 770S Soybean phytoestrogen intake and cancer risk; Adlercreutz CH et al.; Because many Western diseases are hormone-dependent cancers, we have postulated that the Western diet, compared with a vegetarian or semi-vegetarian diet, may alter hormone production, metabolism or action at the cellular level . Recently, our interest has been focused on the cancer-protective role of some hormone-like diphenolic phytoestrogens of dietary origin, the lignans and isoflavonoids . The precursors of the biologically active compounds originate in soybean products (mainly isoflavonoids but also lignans), as well as whole grain cereals, seeds, probably berries and nuts (mainly lignans) . The plant lignan and isoflavonoid glycosides are converted by intestinal bacteria to hormone-like compounds with weak estrogenic and antioxidative activity; they have now been shown to influence not only sex hormone metabolism and biological activity but also intracellular enzymes, protein synthesis, growth factor action, malignant cell proliferation, differentiation and angiogenesis, making them strong candidates for a role as natural cancer protective compounds . Epidemiological investigations support this hypothesis, because the highest levels of these compounds are found in countries or regions with low cancer incidence . This report is a review of results that suggest that the diphenolic isoflavonoids and lignans are natural cancer-protective compounds. J Bacteriol, 1995 Mar, 177(6), 1570 - 5 Characterization of nifB, nifS, and nifU genes in the cyanobacterium Anabaena variabilis: NifB is required for the vanadium-dependent nitrogenase; Lyons EM et al.; Anabaena variabilis ATCC 29413 is a heterotrophic, nitrogen-fixing cyanobacterium containing both a Mo-dependent nitrogenase encoded by the nif genes and V-dependent nitrogenase encoded by the vnf genes . The nifB, nifS, and nifU genes of A . variabilis were cloned, mapped, and partially sequenced . The fdxN gene was between nifB and nifS . Growth and acetylene reduction assays using wild-type and mutant strains indicated that the nifB product (NifB) was required for nitrogen fixation not only by the enzyme encoded by the nif genes but also by the enzyme encoded by the vnf genes . Neither NifS nor NifU was essential for nitrogen fixation in A . variabilis. J Bacteriol, 1995 Mar, 177(6), 1520 - 6 A short-filament mutant of Anabaena sp . strain PCC 7120 that fragments in nitrogen-deficient medium; Bauer CC et al.; Strain 129 is a fragmentation mutant of the filamentous cyanobacterium Anabaena sp . strain PCC 7120 . Growing with fixed nitrogen, this mutant forms filaments that are much shorter than wild-type filaments . Following starvation for fixed nitrogen, strain 129 becomes nearly unicellular and forms few heterocysts, although electron microscopy suggests that proheterocysts form while fragmentation occurs . Starvation for sulfate, phosphate, iron, and calcium does not cause this fragmentation . The affected gene in strain 129, fraC, was cloned by complementation and characterized . It encodes a unique 179-amino-acid protein rich in phenylalanine . Insertional inactivation of the chromosomal copy of fraC results in a phenotype identical to that of strain 129, while complementation using a truncated version of FraC results in only partial complementation of the original mutant . Heterocysts could be induced to form in N-replete cultures of strain 129, as in wild-type cells, by supplying extra copies of the hetR gene on a plasmid . Thus, FraC is required for the integrity of cell junctions in general but is apparently not directly involved in normal differentiation and nitrogen fixation. Mutat Res, 1995 Mar, 327(1-2), 179 - 200 International Commission for Protection against Environmental Mutagens and Carcinogens . An evaluation of the genetic toxicity of paracetamol; Rannug U et al.; During the last years, several reports have indicated genotoxic effects of paracetamol, a widely used non-prescription analgesic and antipyretic drug . Thus, a careful evaluation of a possible genotoxic effect related to paracetamol use is warranted . Studies in vitro and in vivo indicate that the reactive metabolite of paracetamol can bind irreversibly to DNA and cause DNA strand breaks . Paracetamol inhibits both replicative DNA synthesis and DNA repair synthesis in vitro and in experimental animals . Paracetamol does not cause gene mutations, either in bacteria or in mammalian cells . On the other hand, a co-mutagenic effect of paracetamol has been reported . Furthermore, paracetamol increases the frequency of chromosomal damage in mammalian cell lines, isolated human lymphocytes and experimental animals . Two independent studies have shown an increase in chromosomal damage in lymphocytes of human volunteers after intake of therapeutic doses of paracetamol, whereas a third study was negative . Paracetamol-induced chromosomal damage appears to be caused by an inhibition of ribonucleotide reductase . This indicates that a threshold level for the paracetamol-induced chromosomal damage may exist . Genotoxic effects of paracetamol have, however, been demonstrated both in vitro and in vivo at or near therapeutic concentrations . The data indicate that the use of paracetamol may contribute to an increase in the total burden of genotoxic damage in man . Thus, there may be a need to evaluate the therapeutic benefit of paracetamol, taking into consideration not only its potential to induce acute and chronic organ damage, but also genotoxic effects. J Immunol, 1995 Mar 1, 154(5), 2051 - 62 Early appearance of "natural" mucosal IgA responses and germinal centers in suckling mice developing in the absence of maternal antibodies; Kramer DR et al.; We have examined the role of passively transferred maternal Abs in the ontogeny of "natural" mucosal IgA responses before weaning of suckling mice by comparing the immune status of gut-associated lymphoid tissue (GALT) (Peyer's patches, mesenteric lymph nodes, and lamina propria) in 7- to 25-day-old F1 severe combined immunodeficient (scid)/+ mice generated through reciprocal crosses of C.B17 scid/scid and normal congenic (+/+) adult mice . We have also examined the ability of prenatal vs postnatal transfer of maternal immunity to forestall the development of natural neonatal mucosal IgA responses by swapping litters of F1 scid/+ pups at birth between +/+ and scid/scid mothers . Our results demonstrate that F1 scid/+ pups born to or nursed by scid/scid mothers undergo an accelerated development of natural IgA responses that include germinal center reactions in both Peyer's patches and mesenteric lymph nodes . These early IgA responses are evident as: 1) increased frequencies of IgA-producing GALT organ cultures; 2) increased mean IgA output by GALT organ cultures; 3) increased frequencies (> 1 log) of IgA-secreting cells from GALT detected by ELISPOT at 16 days of age; and 4) germinal center development by 17 days of age detected by in vivo bromodeoxyuridine incorporation . Finally, FACS analyses of enteric bacteria isolated from F1 scid/+ pups and stained for the presence of surface-bound mouse IgA demonstrate that the bacterial flora is a major target of both maternal secretory IgA and of the earliest IgA Abs produced in the neonatal GALT of pups deprived of maternal immunity. Infect Immun, 1995 Mar, 63(3), 1130 - 3 Interleukin-10 downregulates protective immunity to Brucella abortus; Fernandes DM et al.; In vivo neutralization of interleukin-10 (IL-10) with an anti-IL-10 monoclonal antibody resulted in up to 10-fold fewer bacteria in the spleens of BALB/c mice infected with the virulent Brucella abortus strain 2308 . In vitro neutralization of endogenous IL-10 in brucella antigen-stimulated cultures of splenocytes from infected mice resulted in increased gamma interferon production in these cultures, whereas exogenous recombinant IL-10 inhibited the ability of peptone-elicited peritoneal macrophages to control intracellular brucellae . These data suggest that IL-10 may be downregulating the immune response to B . abortus by affecting both macrophage effector function and the production of the protective Th1 cytokine gamma interferon. Mol Cell Biol, 1995 Mar, 15(3), 1353 - 63 Homotypic interactions of chicken GATA-1 can mediate transcriptional activation; Yang HY et al.; We used a one-hybrid system to replace precisely the finger II chicken GATA-1 DNA-binding domain with the binding domain of bacterial repressor protein LexA . The LexA DNA-binding domain lacks amino acids that function for transcriptional activation, nuclear localization, or protein dimerization . This allowed us to analyze activities of GATA-1 sequences distinct from DNA binding . We found that strong transcriptional activating sequences that function independently of finger II are present in GATA-1 . Sequences including finger I contain an independent nuclear localizing function . Our data are consistent with cooperative binding of two LexA-GATA-1 hybrid proteins on a palindromic operator . The sensitivity of our transcription assay provides the first evidence that GATA-1 can make homotypic interactions in vivo . The ability of a non-DNA-binding form of GATA-1 to activate gene expression by targeting to a bound GATA-1 derivative further supports the notion that GATA-1-GATA-1 interactions may have functional consequences . A coimmunoprecipitation assay was used to demonstrate that GATA-1 multimeric complexes form in solution by protein-protein interaction . The novel ability of GATA-1 to interact homotypically may be important for the formation of higher-order structures among distant regulatory elements that share binding sites for this transcription factor . We also used the system to test the ability of GATA-1 to interact heterotypically with other activators. Blood, 1995 Mar 1, 85(5), 1341 - 7 Downregulation of proinflammatory cytokine release in whole blood from septic patients; Ertel W et al.; Using animal models or healthy volunteers, injection of lipopolysaccharide (LPS) or bacteria