Ann Intern Med, 1986 Nov, 105(5), 690 - 4 Plesiomonas enteric infections in the United States; Holmberg SD et al.; Thirty-one persons nationwide from whom Plesiomonas shigelloides was isolated in large numbers from stool in 1984 were compared with 62 matched control subjects . Infection with P . shigelloides was strongly associated with eating uncooked shellfish, usually raw oysters, in the 48 hours before the onset of illness (p less than 0.00001) and with foreign travel (p less than 0.00006), usually to Mexico . Most ill persons had self-limited diarrhea with blood and mucus in stool and other clinical findings that suggested enteroinvasiveness of infecting organisms . Two patients developed their illnesses after taking ampicillin for reasons unrelated to diarrhea; plesiomonads recovered from their stools were resistant to ampicillin . Seven persons with gastrointestinal complaints had alleviation or resolution of their symptoms after taking antimicrobial agents to which recovered plesiomonads were susceptible . These findings suggest that P . shigelloides may cause enteric disease in the normal host, that it may be acquired from eating uncooked shellfish, and that it may be a cause of travelers' diarrhea. Zh Mikrobiol Epidemiol Immunobiol, 1986 Nov, (11), 46 - 51 {Phenomenology and molecular mechanisms of Shigella sonnei dissociation: its variability in model in vivo and in vitro systems}; Kaminskii GD et al.; The dynamics of the accumulation of phase II S . sonnei in the infectious process in germ-free rats, during cultivation and in subcultures was studied . The in vivo variability of S . sonnei showed a sharply defined phasic character of the process with the periods of the apparent absence of phase II, the increase of its occurrence and stabilization with the preservation of S-forms in minor amounts . Kanamycin-resistant phase II accumulated in vivo more rapidly than in vitro . Cultivation with sodium desoxycholate was found to accelerate the process of disassociation . The characteristic feature of all dissociants detected in all systems was their plasmid profile: in phase I, plasmids of 120 and 60 Md, as well as small plasmids, were detected; in phase II disassociants, plasmid with a molecular weight of 120 Md was absent . The restriction analysis of total plasmid DNA by means of restrictase EcoR1 showed the absence of 3 fragments in phase II while other 13 fragments were retained . These results open prospects for the molecular cloning of the gene antigen of phase II. Zh Mikrobiol Epidemiol Immunobiol, 1986 Nov, (11), 42 - 6 {Manifestations of genotypic and phenotypic heterogeneity in populations of Shigella sonnei and of humans at different phases of the development of the epidemic process}; Shapiro MI et al.; The study of the structure and developmental dynamics of S . sonnei populations and the state of local immunity to this infection in children under school age has revealed that the interaction of the heterogeneous populations of the parasite and the host is the internal motive force of the self-regulating phasic development of the epidemic process . In the course of this process complementary changes in the virulence of the infective agent and in the immunoresistance of children are generated on the basis of negative feed-back. J Bacteriol, 1986 Nov, 168(2), 762 - 8 In vitro packaging of heteroduplex bacteriophage T7 DNA: evidence for repair of mismatched bases; Masker W; Heteroduplex DNA molecules that were wild type or contained combinations of amber, missense, and temperature-sensitive mutations were prepared from bacteriophage T7 . These DNA molecules were then encapsulated in in vitro packaging reactions so as to produce infective T7 phage . The genotypes of the phage were examined to determine the degree to which mismatched base pairs in the heteroduplex had been corrected . The data show that conversion of the mismatches took place either during in vitro packaging or immediately after infection of either an Escherichia coli or Shigella sonnei host . The mode of mismatch conversion observed in these experiments was independent of the host mutH, mutL, mutS, and uvrD genes . There was no significant amount of discrimination between markers on either of the two complementary strands . The observed frequency of conversion of a mismatch depended on the genetic marker being monitored and on experimental conditions but was generally in the range between 5 and 30%. Ann Inst Pasteur Microbiol, 1986 Nov-Dec, 137B(3), 291 - 5 Elimination of a virulence plasmid from Shigella sonnei and Escherichia coli by antibiotics; Michel-Briand Y et al.; Plasmid pWR105 is a non-self-conjugative plasmid conferring enteroinvasive properties on Shigella sonnei . Loss of this plasmid is accompanied by loss of the invasive phenotype as well as of the form I antigen expression . Thirteen antibiotics belonging to different chemical families were used at subinhibitory concentrations to eliminate pWR105 from S . sonnei and Escherichia coli K12 . Rifampicin, novobiocin, chloramphenicol, cotrimoxazole and erythromycin eliminated the plasmid from both strains . Clindamycin eliminated pWR105 from E . coli only . Several other antibiotics gave a low rate of cure (ciprofloxacin, nalidixic acid, oxolinic acid, nifurzide, tilbroquinol, minocycline) . We may expect that these antibiotics would destabilize plasmids from other Shigella species and enteroinvasive E . coli, as these extrachromosomal DNA molecules share a high degree of homology. Infect Immun, 1986 Nov, 54(2), 435 - 43 Characterization of Shigella flexneri sequences encoding congo red binding (crb): conservation of multiple crb sequences and role of IS1 in loss of the Crb+ phenotype; Daskaleros PA et al.; The ability to bind Congo red (Crb+) is associated with virulence of Shigella flexneri and is encoded by a large, 220-kilobase plasmid . We cloned fragments of this plasmid to isolate the sequences encoding Congo red binding, to determine the degree of conservation of these sequences among S . flexneri strains, and to study the molecular basis for loss of the Crb+ phenotype . At least two separate BamHI fragments cloned into plasmid vectors encode Congo red binding in E . coli or S . flexneri . One Crb+ clone, pTKS2, contains a copy of IS1 adjacent to the crb sequences . IS1 appears to be responsible for deletions leading to loss of Congo red binding in this clone . In addition, this clone was found to integrate into the chromosome at relatively high frequency . Integration resulted in loss of the Crb+ phenotype . A second clone, pTKS15, which has only limited homology to pTKS2, also encodes Congo red binding . The Crb+ phenotype of transformants carrying pTKS15 was detected at 37 degrees C but not at 30 degrees C, and thus it resembles Congo red binding in wild-type S . flexneri . HindIII digests of plasmid DNA from 10 different S . flexneri strains were hybridized to both of these Crb+ clones and to an IS1 probe . More than one fragment hybridized to pTKS2 or pTKS15 . In general, the sizes of these fragments were the same in S . flexneri strains of different serotypes, indicating conservation of these sequences . Three of five copies of IS1 were also found on the large S . flexneri plasmids . Two of the copies were on fragments of the same size in each strain . Analysis of Crb- derivatives of the 10 strains indicated that, although IS1 may be closely linked to crb sequences on the 220-kilobase plasmid, it is not responsible for the majority of deletions of this plasmid associated with loss of Congo red binding. Infect Immun, 1986 Nov, 54(2), 395 - 402 DNA sequence and product analysis of the virF locus responsible for congo red binding and cell invasion in Shigella flexneri 2a; Sakai T et al.; The DNA sequence of virF, a locus associated with virulence and the ability to bind Congo red in Shigella flexneri 2a that is located on a 140-megadalton (230-kilobase) plasmid, was determined and analyzed . It was rich in A and T . The direction of transcription of virF was determined by using a chloramphenicol resistance cartridge . An open reading frame readable in this direction was found . Three proteins, 30, 27, and 21 kilodaltons, all corresponding to those predicted from the above sequence, were produced in minicells containing the virF locus . The three proteins were expressed only weakly in minicells with the 230-kilobase plasmid. J Infect Dis, 1986 Nov, 154(5), 742 - 51 Colonization, virulence, and mucosal interaction of an enteropathogenic Escherichia coli (strain RDEC-1) expressing shigella somatic antigen in the rabbit intestine; Inman LR et al.; The group factor 3,4 somatic antigen was transferred by recombination from a donor strain of Shigella flexneri type 2a to the Escherichia coli O15 rabbit pathogen strain RDEC-1 . A hybrid clone (7482-1-1) that expressed only the S . flexneri 3,4 somatic antigen and a second clone (7482-1-7) from the same mating mixture that expressed only the O15 somatic antigen were compared for virulence in rabbits . The 7482-1-1 strain produced diarrhea in 10% of rabbits versus 84% for the 7482-1-7 strain (P less than .001) . In fluorescent antibody-stained, frozen, sectioned tissues, fewer 7482-1-1 bacteria adhered to ileum (P = .013) and cecum (P = .044) . Light and electron microscopic studies demonstrated that the recombinant strain, which adhered to membranous (M) cells to the same degree as the 7482-1-7 strain, was found beneath the epithelium, where its presence resulted in a marked acute inflammatory response . Both the 7482-1-1 and 7482-1-7 strains exhibited characteristic close adherence to M cells and absorptive ileal and cecal mucosa. J Biol Chem, 1986 Oct 25, 261(30), 13928 - 31 Complete amino acid sequence of Shigella toxin B-chain . A novel polypeptide containing 69 amino acids and one disulfide bridge; Seidah NG et al.; The complete amino acid sequence of the B-chain of Shigella toxin has been determined using both liquid- and gas-phase sequenators . It reveals a 69-amino acid peptide with a single disulfide bridge, predicting a subunit molecular weight of 7691 . No Asn-X-Ser(Thr) sequence was found, confirming the absence of potential N-glycosylation sites . A computer data bank search using a mutation data matrix did not detect any similarity greater than 30% with known sequences to date, indicating a novel primary structure . However, some distant homology with the 103-residue B-chain of cholera and Escherichia coli enterotoxins was revealed . Hydropathy, fractional exposure, and Chou and Fasman calculations all point to an ordered structure with a hydrophobic core spanning residues 36-52 and a hydrophilic domain between residues 10 and 20, the latter probably representing the most antigenic domain. Zh Mikrobiol Epidemiol Immunobiol, 1986 Oct, (10), 76 - 80 {Characteristics of erythrocyte diagnostic agents studied using optical methods}; Panina LK et al.; The quantitative characterization of erythrocyte diagnosticums (ED) has been made by optical methods (light microscopy with the use of an image analyzer, model Magiscan 2, and the opacity spectrum technique) . The following parameters of ED have been determined: the average of the major axis (5.25 +/- 0.57 micron for ED from Shigella sonnei and 5.53 +/- 0.50 micron for ED from Shigella flexneri), the ratio of semiaxes (p approximately equal to 3), the major axis length distribution, the refractive index (1.076 +/- 0.002) . For controlling the concentration of ED the use of the opacity spectrum technique is recommended. Mol Gen Mikrobiol Virusol, 1986 Oct, (10), 16 - 21 {Functional characteristics of plasmids of Shigella sonnei 47 strains}; Kariagina AS et al.; The phenotypic characteristics of Shigella sonnei strain 47 containing 7 plasmids of low molecular weight and 2 plasmids 60-100 Md large have been studied . The strains of Escherichia coli containing the single plasmids or plasmid groups from Shigella sonnei have been obtained by transformation and conjugation . The comparison of phenotypes of the obtained strains has helped to find the plasmid location of the determinants for streptomycin resistance (P7), genes for colicinogenicity and colicin immunity (P5), the enzymes of host cell specificity system Sso47I (P6), Sso47II (P4), and the genes for the conjugative DNA transfer (P9) . Escherichia coli strains producing individual restriction enzymes SsoI and SsoII have been isolated. Am J Pathol, 1986 Oct, 125(1), 69 - 80 Morphologic evaluation of the effects of Shiga toxin and E coli Shiga-like toxin on the rabbit intestine; Keenan KP et al.; The effects of a Shiga toxin derived from Shigella dysenteriae Type 1, Strain 60R, and a Shiga-like toxin from the enterohemorrhagic Escherichia coli O157:H7, Strain 933, were studied in the in vivo rabbit ileal loop model . The effects of both toxins were similar and resulted in severe villus blunting by 18-24 hours after exposure . With both toxins, a dose effect was noted; and the lesions, first detected at 2 hours after inoculation, became more severe over time . Both toxins appeared to act directly and selectively on the mature columnar absorptive epithelium of the intestinal villus, which resulted in the premature expulsion of these cells from the lateral villus wall, with a decrease in the villus/crypt ratio . The goblet mucous cells remained attached and frequently formed clusters on the blunt villus apices . The crypt epithelium underwent a rapid proliferation and maintained the epithelial integrity . The ultrastructural changes observed in the toxin-injured villus absorptive cells suggested that these cells underwent a process of apoptosis, rather than necrosis . These findings suggest that both toxins act in vivo in the small intestine on a specific cell population, the mature, differentiated absorptive villus epithelium. Immunol Lett, 1986 Oct, 13(4), 197 - 201 Lack of serologically defined arthritogenic Shigella flexneri cell envelope antigens in post-dysenteric arthritis; van Bohemen CG et al.; Post-dysenteric or reactive arthritis (ReA) is closely associated with HLA-B27 . This histocompatibility antigen is heterogeneous and consists of 2 serologically defined variants: B27M1+M2+ and B27M1+M2- . This paper gives a qualitative evaluation of the antibodies present in the sera of 62 patients with dysentery due to Shigella flexneri 2a, a known arthritogenic bacterium . The patients were classified in 4 groups: B27M1+M2+ReA+ (n = 5), B27M1+M2+ReA- (n = 7); B27M1+M2-ReA- (n = 1); B27-ReA- (n = 49) . The isolated infectant possessed cell envelope antigens with B27M2-like epitopes (Mr 20,000) . Analysis of the spectrum of antibodies directed against the separated cell envelope antigens of S . flexneri in the sera of these patients revealed 7 main patterns of reactivity . The detectable immunogens encompassed protein stainable antigens (Mr 98, 78, 68, 54, 50, 44, 41, 35, 14 and 13 kDa), lipopolysaccharides and peptidoglycan . None of the sera possessed detectable antibodies to the B27M2-like antigen . Consequently, this antigen is unlikely to be associated with ReA, and this applies equally to other antigens or patterns of antigens . The arthritogenicity of S . flexneri may therefore not be determined by the presence or absence of detectable antibody titers to certain cell envelope antigens . We hypothesize that other properties of these antigens could be of significance. J Immunol, 1986 Oct 1, 137(7), 2361 - 6 Characterization of Shigella flexneri-specific murine monoclonal antibodies by chemically defined glycoconjugates; Carlin NI et al.; Chemically defined glycoconjugates are demonstrated to have considerable potential for selecting hybridoma antibodies directed toward O-antigenic determinants, especially when used in combination with a panel of well-characterized LPS molecules . Monoclonal antibodies specific for the Shigella flexneri O-antigens of serogroup 5b, variants X and Y, were generated after immunization of BALB/c mice with killed bacterial cells, and active hybrids were selected on the basis of ELISA performed with the purified serotype-specific LPS antigen . Subsequent screening with a variety of glycoconjugates, derived from synthetic oligosaccharides and larger structures obtained by phage Sf6/endo-rhamnosidase hydrolysis of purified LPS established a detailed profile of binding characteristics for Shigella flexneri variant Y-specific antibodies . Together with the results of precipitin analysis and heavy chain isotyping experiments, a limited number of antibodies were selected as candidates for detailed studies of the antibody combining site. Microbiologia, 1986 Oct, 2(2), 81 - 8 Plasmid profile analysis as a tool for characterization of epidemic Shigella sonnei strains; Mendoza MC et al.; The usefulness of plasmid profile analysis as a marker to determine the strain number of Shigella sonnei implicated in an epidemic outbreak and its prevalency in our community was studied . We could define two multirresistant strains and determine that the drug resistance was plasmid mediated in both . One strain carried a tra- plasmid which encoded (Sm, Sd)r, the other one carried two tra+ plasmids which encoded (Ap-Cb, Sm, Sd)r and (Sm, Km-Nm, Sd, Tp)r respectively . These strains and some coisolates collected during the later phase of the outbreak carried cryptic plasmids which showed similar electrophoretic mobilities. Am J Gastroenterol, 1986 Oct, 81(10), 944 - 50 Cytomegalovirus infection of the alimentary tract: a clinicopathological correlation; Hinnant KL et al.; Alimentary tract cytomegalovirus (CMV) infections of 24 patients were reviewed, including 19 with the acquired immune deficiency syndrome . CMV inclusion bodies (CMV-IB) were calibrated per mm2 of tissue . CMV-IB counts were correlated with biopsy site, inflammatory response, and clinical parameters . Colonic biopsies showed the highest counts . Biopsies of the right colon had about three times as many CMV-IBs as those of the left . Upper alimentary tract biopsies had low counts . Mesenchymal cells were most affected (97%); 35% were identified as endothelial and 6% as smooth muscle . Only 3% of CMV-IBs were in epithelial cells . Grades of inflammation, 1-5, correlated directly with CMV-IB counts up to grade 4 . In grade 5 inflammation tissue destruction was so severe that CMV-IBs were difficult to recognize . Ulcers were demonstrated in more than half of all patients, either histologically or endoscopically . The inflammatory response was nonspecific, except for patchy infiltrates and the absence of lymphoid follicles, crypt abscesses, or granulomas . Gastrointestinal infections, such as shigellosis, candidiasis, mycobacteriosis, and cryptosporidiosis coexisted in 17 patients . No correlation was found between CMV-IB counts and severity of symptoms or length of survival . Alimentary tract CMV infections was the first manifestation of the acquired immune deficiency syndrome in 11 patients . Survival ranged from 2 wk to 19 months. Infect Immun, 1986 Oct, 54(1), 32 - 6 Isolation, characterization, and mapping of Tn5 insertions into the 140-megadalton invasion plasmid defective in the mouse Sereny test in Shigella flexneri 2a; Sasakawa C et al.; Using Shigella flexneri 2a YSH6000, we isolated 304 independent Tn5 insertion mutants in the 230-kilobase invasion plasmid, pMYSH6000 . The site of each Tn5 insertion was assigned to 23 SalI fragments on the previously made SalI cleavage map of pMYSH6000 . Among the 304 insertions, 150 were negative in expression of four phenotypes examined (mouse Sereny test {Ser}, invasion into epithelial cells {Inv}, Congo red binding {Pcr}, and inhibition of bacterial growth {Igr} ): 12 were Ser- Inv+ Pcr+ Igr+, and 142 were positive in all four phenotypes . Tn5 insertions in the avirulent mutants were distributed in two separate SalI fragments, F and G, and in four contiguous SalI fragments, B, P, H, and D . Fragment G contains a novel class of determinant(s) which is required only for Ser+ but not for Inv+, Pcr+, and Igr+ . Fragment F contains the previously characterized virF locus . B, P, H, and D each contained both virulent and avirulent Tn5 insertions . This indicates that more than two gene clusters exist within this region . Both are required for expression of all four virulence phenotypes. Scand J Prim Health Care, 1986 Sep, 4(3), 183 - 90 Growth, nutritional status and infant mortality of Turkish immigrant preschool children; Mjones S et al.; Turkish preschool children in Stockholm, Sweden, 59 born 1979, 32 born 1976 and 43 siblings of preschool age have been studied as to nutritional status including anthropometric parameters, dental caries, haemoglobin level and bacteriological and parasitological findings in faeces and as to deaths of siblings in Turkey and Sweden . Growth was very close to that of the Swedish standard . Two of three children with slightly subnormal growth had had recent direct or indirect contact with the home country . There were no signs of nutritional deficiency but dental caries was common . There was little anaemia . One case of shigellosis was found . Mortality of infancy was significantly reduced after immigration . The group had made a successful nutritional adaptation in Sweden considering their origin in regions of Turkey with a high prevalence of growth failure . Health care in this group should aim at prevention of diarrhoeal disease on visits to Turkey as well as of dental caries. Acta Paediatr Scand, 1986 Sep, 75(5), 713 - 8 Observations questioning a protective role for breast-feeding in severe rotavirus diarrhea; Glass RI et al.; To investigate whether breast-feeding protects children against rotavirus diarrhea (RVD), we compared rates of breast-feeding by age and enteric pathogens among 2,276 children with diarrhea 0-4 years of age who attended a diarrhea hospital in Bangladesh . Infants 0-5 months were less likely to be breast-fed than children 6-11 months of age suggesting that some protection against diarrhea with all agents was associated with early breast-feeding . In every age group studied, breast-feeding was more common among children with RVD than among children with non-RVD whereas it was less common among children with cholera and shigellosis . Twenty percent of breast milks consumed by infants less than 1 year of age had high levels of neutralizing activity (greater than or equal to 320) to the Wa strain of rotavirus but this activity did not appear to be protective since the 30 infants with RVD consumed milk which had titers that did not differ significantly from those consumed by 44 infants with diarrhea of other cause . Despite the prolonged breast-feeding which is common in Bangladesh, the mean age of hospitalization with RVD is approximately the same as in countries where the duration of breast-feeding is quite short . None of these 3 independent observations support a protective role for breast-feeding against rotavirus diarrhea after the first months of life. J Clin Microbiol, 1986 Sep, 24(3), 498 - 500 Comparison of DNA probes and the Sereny test for identification of invasive Shigella and Escherichia coli strains; Wood PK et al.; Forty-two Shigella and 29 Escherichia coli strains were screened for invasiveness in the Sereny test and for hybridization with two recently described DNA probes for the invasiveness plasmid . Both probes produced identical results . All Sereny-positive strains hybridized with both DNA probes . Three Sereny-negative strains also hybridized with the probes, suggesting that there are strains containing the invasiveness plasmid that are not pathogenic in animal models. Bioorg Khim, 1986 Sep, 12(9), 1240 - 52 {Somatic antigens of Shigella and Escherichia coli . Determination of the structure of O-specific polysaccharide from Shigella boydii type 12 and its serological properties compared with the O-specific polysaccharide from Escherichia coli}; L'vov VL et al.; The structure of the O-specific polysaccharide of the somatic antigen (lipopolysaccharide) of Shigella boydii, type 12, was established by 1H- and 13C-NMR, methylation analysis and partial acid hydrolysis methods . The polysaccharide consists of pentasaccharide repeating units of the following structure: (formula; see text) The amount of O-acetyl groups was far less than stoichiometric, only about 2 for 3-4 repeating units . Nevertheless, the results of serological studies revealed 3-O-acetyl-alpha-L-rhamnose residue to be the major immunodominant group . In spite of the presence of similar trisaccharide fragments, the lipopolysaccharide and polysaccharide from Shigella boydii type 12 gave no crossreaction with lipopolysaccharide and polysaccharide from Escherichia coli 07 . The possible reasons of the absence of serological relatedness between the Sh . boydii, type 12, and E . coli 07 cells were discussed. Bioorg Khim, 1986 Sep, 12(9), 1213 - 24 {Synthesis of a repeating unit and the dimer of the repeating unit of the major chain of Shigella flexneri O-antigen polysaccharide}; Tsvetkov IuE et al.; Methyl glycoside of the tetrasaccharide GlcNAc(beta 1-2)Rha(alpha 1-2)Rha(alpha 1-3)Rha, which represents a repeating unit of the basic chain of Shigella flexneri O-antigenic polysaccharides, was synthesized using acylated monosaccharide synthons . A dimer of the repeating unit, octasaccharide {GlcNAc(beta 1-2)Rha(alpha 1-2) Rha(alpha 1-3)Rha(alpha 1-3)}2-OMe was obtained by TrClO4-catalyzed condensation of two tetrasaccharide blocks. Cell, 1986 Aug 15, 46(4), 551 - 5 A genetic determinant required for continuous reinfection of adjacent cells on large plasmid in S . flexneri 2a; Makino S et al.; We have identified a region (virG) on the 230 kb virulence plasmid of S . flexneri that is required for cell-to-cell spread of the bacterium . Tn5 insertions into this region result in avirulent mutants that can initially invade and multiply in epithelial cells, but tend to lose active movement and tend to localize within the cytoplasm, where they are gradually extinguished without infecting adjacent cells . The virG region was localized to within 4 kb and may contain a single cistron . Sequences hybridizing to this region were found in all intact virulence plasmids of Shigellae and enteroinvasive E . coli. Infect Immun, 1986 Aug, 53(2), 372 - 7 Pathogenesis of Shigella diarrhea: rabbit intestinal cell microvillus membrane binding site for Shigella toxin; Fuchs G et al.; This study examined the binding of purified 125I-labeled shigella toxin to rabbit jejunal microvillus membranes (MVMs) . Toxin binding was concentration dependent, saturable, reversible, and specifically inhibited by unlabeled toxin . The calculated number of toxin molecules bound at 4 degrees C was 7.9 X 10(10) (3 X 10(10) to 2 X 10(11))/micrograms of MVM protein or 1.2 X 10(6) per enterocyte . Scatchard analysis showed the binding site to be of a single class with an equilibrium association constant, K, of 4.7 X 10(9) M-1 at 4 degrees C . Binding was inversely related to the temperature of incubation . A total of 80% of the labeled toxin binding at 4 degrees C dissociated from MVM when the temperature was raised to 37 degrees C, but reassociated when the temperature was again brought to 4 degrees C . There was no structural or functional change of MVM due to toxin as monitored by electron microscopy or assay of MVM sucrase activity . These studies demonstrate a specific binding site for shigella toxin on rabbit MVMs . The physiological relevance of this receptor remains to be determined. Biokhimiia, 1986 Aug, 51(8), 1369 - 76 {Study of the conditions of activation and stabilization of DNA-methylases from Shigella sonnei 47 during fractionation, purification and storage}; Suchkov SV et al.; A comparative study of activation factors and stabilization conditions of partially purified and individual fractions of DNA-methylases of Shigella sonnei 47 was carried out . The stability of DNA-methylases in the course of storage was examined . The influence of activating factors and stabilization conditions differed significantly depending on the degree of purification and composition of methylase preparations . It was shown that glycerol is ineffective as a stabilizing agent . The activating effect of Ca2+ on Shigella sonnei 47 DNA-methylases was found to be universal, while albumin was shown to exert a more potent stabilizing action . The inactivating effect of proteases on DNA methylation enzymes during storage was demonstrated . A phenomenon of spontaneous fluctuations in the methylating activity of enzymatic preparations of Shigella sonnei 47 upon storage was observed. Zh Mikrobiol Epidemiol Immunobiol, 1986 Aug, (8), 10 - 3 {Detection of immunosuppression factors in Shigella sonnei}; Frolov AF et al.; A factor, making noninvasive shigellae and other bacteria capable of suppressing immunological memory and secondary immune response manifested as delayed hypersensitivity, has been detected in the germ-free filtrates of the broth cultures of invasive S . sonnei. South Med J, 1986 Aug, 79(8), 1046 - 7 Shigellosis complicated by acute appendicitis; Lending RE et al.; Acute appendicitis developed in a 23-year-old Mexican-American man with acute bacillary dysentery (shigellosis) . This is the first such case reported in the English literature. Am J Epidemiol, 1986 Aug, 124(2), 299 - 305 Contaminated produce--a common source for two outbreaks of Shigella gastroenteritis; Martin DL et al.; Simultaneous outbreaks of Shigella sonnei gastroenteritis occurred in October 1983 at two Texas university campuses 60 miles (96 km) apart . There were no common food handlers, recreational activities, water sources or swimming areas to explain the introduction of Shigella at both campuses . However, tossed salads were found to be associated with illness at both campuses . The investigation disclosed that both schools had received produce shipments from the same company during the week preceding these outbreaks . Shigella isolates from cases at both universities, sent to the Centers for Disease Control for plasmid analysis and colicin typing, were found to be identical . The same organism was evidently not a frequent cause of shigellosis within a 160-mile (256 km) radius of these universities since only 19% of control isolates chosen from this area were identical to the type which caused these outbreaks . This is the first report of two related outbreaks of shigellosis that were caused by a contaminated food source and not by a food handler . Simultaneous foodborne outbreaks of shigellosis should trigger a search for potential contamination at every step of food handling from farm to kitchen. Microb Pathog, 1986 Aug, 1(4), 325 - 34 Shiga toxin from Shigella dysenteriae 1 inhibits protein synthesis in reticulocyte lysates by inactivation of aminoacyl-tRNA binding; Brown JE et al.; Inhibition of the peptide elongation cycle of eukaryotic protein synthesis by Shiga toxin from Shigella dysenteriae 1 was examined in toxin-treated reticulocyte lysate mixtures . Peptidyl transferase activity of toxin-treated ribosomes was measured by following the decrease in peptidyl-tRNA concentrations when puromycin was added after incubation with toxin . Concentrations of {3H}leucine-labeled peptidyl-tRNA were measured by extraction with cetyltrimethylammonium bromide . The data suggest that Shiga toxin inhibited aminoacyl-tRNA binding . Toxin-treated ribosomes retained peptidyl transferase activity, and toxin did not block translocation . Furthermore, no inhibition of initiation of protein synthesis could be observed . Finally, Shiga toxin had no detectable nucleolytic effect on polysomal 28S rRNA, nor was hydrolysis of 5.8S or 5S rRNA observed. Infect Immun, 1986 Aug, 53(2), 352 - 8 Identification of Shigella sonnei form I plasmid genes necessary for cell invasion and their conservation among Shigella species and enteroinvasive Escherichia coli; Watanabe H et al.; A series of Tn1 insertions in pSS120, the 120-megadalton form I plasmid of Shigella sonnei, were constructed by a Tn1-mediated conduction system previously described (H . Watanabe and A . Nakamura, Infect . Immun . 48:260-262, 1985, and screened for cell invasion in a tissue culture assay . The analysis of Tn1 insertion sites of seven noninvasive mutants suggested that four separate HindIII fragments were necessary for cell invasion . HindIII fragments including Tn1 of mutant plasmids were cloned into a vector plasmid, pACYC184 . The DNA was used as a DNA probe to identify the corresponding, parental HindIII fragments . We identified one contiguous molecule of 2.6- and 4.1-kilobase pair (kb) HindIII fragments as being responsible for restoring cell invasiveness to the three mutant plasmids, pHW505, pHW510, and pHW511 . Polypeptide analysis in minicells demonstrated that the contiguous HindIII fragments of 2.6 and 4.1 kb coded for at least four polypeptides, of 38, 41, 47, and 80 kilodaltons (kDa) . A comparison of polypeptides synthesized by parental and mutant plasmids strongly suggested that the 38-kDa protein was essential for cell invasion . The 4.1-kb DNA which encoded the 38-kDa protein was conserved among plasmids of Shigella species and enteroinvasive Escherichia coli. Antibiot Med Biotekhnol, 1986 Aug, 31(8), 600 - 2 {Relationship of the pKMR plasmids detected in Shigella circulating in the Krasnodar Territory to incompatibility groups}; Shishkina ZV et al.; Shigella strains showed that in 43.9 per cent of the strains this feature was controlled by conjugative R plasmids . On the whole these were the plasmids allotting the bacterial cell with resistance simultaneously to SmTcCm (37.1 per cent) and SmTc (17.1 per cent) . The plasmids with other phenotypes were less frequent: SmApCm, 11.4 per cent, TcApCm, 8.6 per cent; SmTcApCm, 8.6 per cent; Tc, 8.6 per cent; Sm, 2.9 per cent; Cm, 2.9 per cent and TcCm, 2.9 per cent . The incompatibility groups of 19 plasmids were determined: Inc I zeta, 28.6 per cent; Inc zeta I and Inc B, 14.2 per cent; Inc FII, 8.6 per cent and Inc I alpha, 2.9 per cent. Immunol Lett, 1986 Aug, 13(1-2), 71 - 4 HLA-B27M1M2 and high immune responsiveness to Shigella flexneri in post-dysenteric arthritis; van Bohemen CG et al.; The heterogeneous HLA-B27 antigen is closely associated with post-infectious or reactive arthritis (ReA) and is comprised of two serologically defined variants: B27M1+M2+ and B27M1+M2- . An outbreak of dysentery (n = 120) caused by a Shigella flexneri 2a strain, which possessed cell envelope antigens with epitopes resembling B27M2, resulted in five B27M1+M2+ patients with ReA . The remaining seven B27M1+M2+, one B27M1+M2- and all but three B27-negative patients remained free of joint symptoms; the latter three displayed arthralgia . IgM, IgG and IgA serum titers were statistically raised in all patient groups, but were exceptionally and persistently high in the B27M1+M2+ patients with ReA, especially IgA, as determined in acute-phase sera and sera sampled 1 year after dysentery . B27M1+M2+ thus appears to be a marker for a subset of disease, characterized by a high immune response . It is concluded that the B27M2 epitope is not unequivocally disease-related to Shigella ReA, that B27M1+M2+ is not likely to be the only immune-response-regulating gene involved in this form of ReA and that cross-reactivity between bacterial antigenic epitopes and B27 can only be part of a multifactorial process leading to ReA and in itself not sufficient to produce ReA . The intensity of the immune response appears to be another important factor. Vet Pathol, 1986 Jul, 23(4), 425 - 30 Generalized amyloidosis in rhesus monkeys; Blanchard JL et al.; Necropsy materials from 57 cases of generalized amyloidosis in rhesus monkeys were reviewed . Clinically, animals with the disease were characterized by cachexia with muscle wasting, recurrent diarrhea, and arthritis . Gross lesions included hepatomegaly, splenomegaly, chronic/active colitis, fibrous strictures of the cecocolic junction, osteoarthritis, and generalized muscle atrophy . Histologic examination revealed minimal to severe deposits of amyloid in the small intestine (100%), spleen (93%), large intestine (67%), liver (40%), lymph nodes (71%), stomach and/or adrenal gland (32%) . More amyloid was deposited in the spleen, liver, and small intestine than in other organs . Shigella sp . were isolated from feces in 23% of the cases and 84% had histologic evidence of colitis . Other findings indicated that 100% of the animals had lung mites, 25% had strictures of the cecocolic junction, and 40% had osteoarthritis . Thirty percent of the cases occurred in animals 10 months to 5 years of age, 10% in ages 6 to 10 years, and 60% in animals greater than 10 years old. Postgrad Med J, 1986 Jul, 62(729), 663 - 4 Septicaemia and pleural effusion due to Plesiomonas shigelloides; Humphreys H et al.; Plesiomonas shigelloides is a rare cause of invasive infection, most clinically significant isolates being from the gastrointestinal tract of animals and man . Recently it has been implicated as an increasing cause of opportunistic infections . We report a case of P . shigelloides septicaemia and pleural effusion in a patient with pre-existing alcoholic liver disease . This case serves to illustrate the possible role of P . shigelloides as an opportunistic pathogen in a compromised host especially where there is co-existing liver disease. Infect Immun, 1986 Jul, 53(1), 57 - 63 Serum immune response to Shigella protein antigens in rhesus monkeys and humans infected with Shigella spp; Oaks EV et al.; The serum antibody response to proteins encoded by the virulence-associated plasmid of Shigella flexneri was determined in monkeys challenged with virulent S . flexneri serotype 2a . With water-extractable antigen in an enzyme-linked immunosorbent assay, a significant increase in antibody titer against proteins from a plasmid-carrying, virulent strain of S . flexneri serotype 5 could be demonstrated in convalescent sera . There were minimal antibody titers against proteins from an avirulent (plasmid-free) organism . Previously identified plasmid-coded polypeptides a, b, c, and d were predominant antigens recognized by a majority of the convalescent sera in immunoblots . An additional 140-megadalton plasmid-coded polypeptide was also recognized by half of the sera . Convalescent serum from an infected monkey recognized antigens on the bacterial surface in several different plasmid-containing Shigella species and in an enteroinvasive Escherichia coli strain . A survey of sera obtained from children 5 to 10 years of age who had been infected with S . flexneri or S . sonnei revealed high enzyme-linked immunosorbent assay titers in both acute and convalescent sera against a water extract from a virulent Shigella strain . In contrast, children under 3 years of age had no antibody titer in either acute or convalescent sera against the virulence-associated shigella proteins, while 3- to 4-year-old children mounted an immune response against these proteins only in convalescence. Rev Infect Dis, 1986 Jul-Aug, 8(4), 539 - 47 Acute infectious diarrhea among children in day care: epidemiology and control; Pickering LK et al.; The incidence of diarrhea among children in day care centers is highest for those under three years of age . Limited studies indicate that diarrhea occurs more frequently among children enrolled at these centers than among age-matched children cared for at home or in family day care . Most reported outbreaks have been caused by rotavirus, Giardia, Shigella, or combinations of these organisms . Children in day care centers commonly excrete enteropathogens in the absence of symptoms; the significance of this phenomenon in transmission is unknown . An association between higher rates of diarrhea and selected characteristics of centers--the most important of which is the presence of non-toilet-trained children--has been shown . The contamination of hands, communal toys, and other classroom objects as well as a lack of infection control measures play a role in the transmission of enteropathogens in outbreaks of diarrhea in day care centers . Spread of infection from non-toilet-trained children in centers to their families is common . Potential ways of dealing with this situation include education; development, implementation, and enforcement of regulations; and use of infection control measures . However, the effectiveness of specific control measures has not been systematically evaluated. J Bacteriol, 1986 Jul, 167(1), 251 - 6 Expression of the unassembled capsid protein during infection of Shigella sonnei by bacteriophage T7 results in DNA damage that is repairable by bacteriophage T3, but not T7, DNA ligase; Beck PJ et al.; The abortive infection of bacteriophage T7 in Shigella sonnei D2 371-48 is characterized by a premature inhibition of phage DNA replication and nucleolytic breakdown of all phage DNA . Mutations in T7 gene 10 which are recessive to the presence of the wild-type allele can alleviate the restriction of phage growth . Phage T3 productively infects S . sonnei D2 371-48, as does a T7-T3 hybrid phage that contains, in particular, a gene 10 of T7 origin . It is the presence of T3 DNA ligase that allows phage growth on S . sonnei D2 371-48, and this enzyme can also rescue wild-type T7 from the abortive infection . T7+ is therefore functionally ligase deficient during the infection of S . sonnei D2 371-48; this deficiency is a result of the expression of the phage capsid protein, but it is independent of the assembly of the protein into a procapsid or other morphogenetic structure. Infect Immun, 1986 Jul, 53(1), 135 - 40 Two toxin-converting phages from Escherichia coli O157:H7 strain 933 encode antigenically distinct toxins with similar biologic activities; Strockbine NA et al.; Escherichia coli O157:H7 strain 933 contains two distinct toxin-converting phages (933J and 933W) . The biologic activities and antigenic relationship between the toxins produced by 933J and 933W lysogens of E . coli K-12, as well as the homology of the genes that encode the two toxins, were examined in this study . The 933J and 933W toxins, like Shiga toxin produced by Shigella dysenteriae type 1, were cytotoxic for the same cell lines, caused paralysis and death in mice, and caused fluid accumulation in rabbit ileal segments . The cytotoxic activity of 933J toxin for HeLa cells was neutralized by anti-Shiga toxin, whereas the activity of 933W toxin was not neutralized by this antiserum . In contrast, an antiserum prepared against E . coli K-12(933W) neutralized 933W toxin but not 933J toxin or Shiga toxin . For E . coli 933, most of the cell-associated cytotoxin was neutralized by anti-Shiga toxin, whereas most of the extracellular cytotoxin was neutralized by anti-933W toxin . However, a mixture of these antisera indicated the presence of both toxins in cell lysates and culture supernatants . Among 50 elevated cytotoxin-producing strains of E . coli, we identified 11 strains isolated from cases of diarrhea, hemorrhagic colitis, or hemolytic uremic syndrome that produced cell-associated cytotoxins which were neutralized by the 933W antitoxin . Southern hybridization studies showed that the cloned toxin structural genes from phage 933J hybridized with DNA from phage 933W under conditions estimated to allow no more than 26% base-pair mismatch . These findings indicate that E . coli produces two genetically related but antigenically distinct cytotoxins with similar biologic activities which we propose to name Shiga-like toxins I and II . Strains of E . coli that produce elevated levels of Shiga-like toxin I or Shiga-like toxin II, or both, have been associated with the clinical syndromes of diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome. Microbiol Sci, 1986 Jul, 3(7), 216 - 9 Microbiological methods and plasmid pattern analysis in epidemiological control of Shigella sonnei infections; Bratoeva MP; Shigella sonnei plays an important aetiological role in bacillary dysentery . Advances in methodology for the study of intraspecies differentiation are proving to be valuable in the epidemiological analysis of infections caused by this agent. Infect Immun, 1986 Jul, 53(1), 110 - 5 Shigella flexneri O-antigen epitopes: chemical and immunochemical analyses reveal that epitopes of type III and group 6 antigens are identical; Carlin NI et al.; In the present investigation we studied the nature of a Shigella flexneri O antigen, the type III antigen . No structural entity has yet been attributed to this antigen . Phenol-water-extracted lipopolysaccharides (LPSs) from all S . flexneri serotypes possessing either group 6 antigen or both type III antigen and group 6 antigen were subjected to de-O-acetylation by weak alkali treatment . Two-dimensional nuclear magnetic resonance spectra of serotype 3a native LPS and de-O-acetylated LPS were completely resolved, confirming the established structure (L . Kenne, B . Lindberg, K . Petersson, E . Katzenellenbogen, and E . Romanowska, Eur . J . Biochem . 91:279-284, 1978) and proving that no other structural alteration was introduced by the treatment . Both native and de-O-acetylated LPSs of serotypes 1b, 3a, 3b, and 4b were tested in an enzyme-linked immunosorbent assay against four absorbed rabbit anti-S . flexneri antisera and three monoclonal antibodies with specificity for different parts of the S . flexneri type III antigen and group 6 antigen . Upon de-O-acetylation, all binding capacity for the tested antibodies was lost . The chemical and immunological results indicate that the type III and group 6 antigens of S . flexneri are one and the same, a result of the presence of the O-acetyl group in rhamnose III of the repeating unit of the S . flexneri O-antigen chain. Infect Immun, 1986 Jul, 53(1), 103 - 9 Monoclonal antibodies specific for Shigella flexneri lipopolysaccharides: clones binding to type I and type III:6,7,8 antigens, group 6 antigen, and a core epitope; Carlin NI et al.; Monoclonal antibodies against the Shigella flexneri lipopolysaccharide (LPS) were generated in two fusions by using the myeloma cell line Sp2/0 as a fusion partner with spleen cells from BALB/c mice immunized with S . flexneri serotypes 1b and 3a bacteria . The antibodies were characterized by immunoblotting, enzyme-linked immunosorbent assay (ELISA), hemagglutination, and coagglutination . Four different types of monoclonal antibodies were isolated: antibodies specific for the core antigen of the LPS, antibodies specific for the type I O antigen, antibodies specific for the group 6 O antigen, and antibodies specific for the type III:6,7,8 O antigen . The core-specific antibodies were shown to be specific for the Escherichia coli R3 core, which all S . flexneri LPSs tested, except for S . flexneri serotype 6 LPS, have . The type I O antigen-specific antibodies were shown to bind exclusively to S . flexneri serotypes 1a and 1b in ELISA . The type III:6,7,8 O-antigen-specific antibodies were specific for S . flexneri serotype 3a in ELISA and hemagglutination . Two different group 6 O-antigen-specific antibodies were bound . One was bound in both ELISA and hemagglutination to LPSs of S . flexneri serotypes 1b, 3a, 3b, and 4b, whereas the second was bound only to LPSs of serotypes 3a, 3b, and 4b in ELISA but to LPSs of all four serotypes in hemagglutination . The specificity of the isolated I, III:6,7,8, and group 6 monoclonal antibodies was verified by coagglutination of 363 S . flexneri clinical isolates. J Exp Med, 1986 Jun 1, 163(6), 1391 - 404 Pathogenesis of shigella diarrhea . XI . Isolation of a shigella toxin-binding glycolipid from rabbit jejunum and HeLa cells and its identification as globotriaosylceramide; Jacewicz M et al.; A glycolipid that specifically binds shigella toxin was isolated from both HeLa cells and rabbit jejunal mucosa and identified as globotriaosylceramide (Gb3) by its identical mobility on HPTLC to authentic erythrocyte Gb3 . Toxin also bound to a band tentatively identified as alpha-hydroxylated Gb3 . In addition, toxin bound to P1 antigen present in group B human erythrocyte glycolipid extracts . The common feature of the three binding glycolipids is a terminal Gal alpha 1----4Gal disaccharide linked beta 1----4 to either Glc or GlcNAc . Globoisotriaosylceramide, which differs from Gb3 only in possessing a Gal alpha 1----3Gal terminal disaccharide, and LacCer, which lacks the terminal Gal residue of Gb3, were incapable of binding the toxin . Binding was shown to be mediated by the B subunit by the use of isolated toxin A and B subunits and monoclonal subunit-specific antibodies . Gb3-containing liposomes competitively inhibited the binding of toxin to HeLa cell monolayers but did not inhibit toxin-induced cytotoxicity . These studies show an identical carbohydrate-specific glycolipid receptor for shigella toxin in gut and in HeLa cells . The toxin B subunit that mediates this binding has also been shown to recognize a glycoprotein receptor with different sugar specificity . Thus, we have demonstrated that the same small (Mr 6,500) B subunit polypeptide has two distinctive carbohydrate-specific binding sites . The Gal alpha 1----4Gal disaccharide of the glycolipid toxin receptor is also recognized by the Gal-Gal pilus of uropathogenic E . coli . This suggests the possibility that the pilus and toxin B subunit contain homologous sequences . If this is true, it may be possible to use the purified Gal-Gal pilus to produce toxin-neutralizing antibodies. Am J Public Health, 1986 Jun, 76(6), 627 - 30 Control of day care shigellosis: a trial of convalescent day care in isolation; Tauxe RV et al.; Simultaneous outbreaks of S . sonnei infections occurred in September 1983 at two day care centers in Seattle, Washington . At both centers, there were high rates of diarrheal illness in the children (22/80 and 11/23, respectively), in staff members, and in family members of the ill children . The smaller center was temporarily closed, but the larger one remained open . Convalescent children and staff whose diarrhea had resolved and who were receiving appropriate antimicrobials were allowed to return to the larger center before negative cultures were obtained and were separated in an isolation room . No culture-proven cases of shigellosis occurred at either center in the following two months . Plasmid profile determinations suggested the Shigella were not transmitted from the center that remained open to the community . Estimated need for alternate child care requirements at the closed center were 100-fold greater than at the center with the isolation room . Caring for convalescent children in isolation at day care centers while they receive antimicrobial therapy may be a useful strategy for controlling day care shigellosis. Microb Pathog, 1986 Jun, 1(3), 307 - 24 Genetic and biochemical analysis of Shigella dysenteriae 1 O antigen polysaccharide biosynthesis in Escherichia coli K-12: structure and functions of the rfb gene cluster; Sturm S et al.; The genetic organization and functions of the Shigella dysenteriae 1 rfb gene cluster, which specifies the somatic O antigen in this organism, have been studied in Escherichia coli K-12 by insertion and deletion mutagenesis of pSS9, a pBR322 hybrid containing the Shigella rfb genes . On the basis of the sensitivity/resistance to rough-specific bacteriophage T3 of E . coli K-12 derivatives containing mutant pSS9 plasmids, of the banding patterns and immunoreactivity of LPS isolated from such derivatives and electrophoresed on SDS-polyacrylamide gels, and of the sugar composition of the polysaccharide portion of the LPS determined by chemical analysis, six determinants for O antigen production were identified and localized . At least two determinants are involved in synthesis of TDP-rhamnose and the transfer of a rhamnose residue to the galactose-substituted core . One of these functions is probably TDP-rhamnose synthetase . A third function effects the transfer of a second rhamnose residue to the rha----gal-substituted core . A fourth function, for which evidence was obtained for two determinants (cistrons), is N-acetylglucosamine transferase, whereas a sixth determinant is necessary for extension of the first completed side chain repeat unit to the full O antigen polymer . These results confirmed the previously-determined chemical composition of the S . dysenteriae 1 O antigen and demonstrated that the order of the sugars is glcNAc----rha----rha----gal with gal as the first sugar linked to the core . Evidence was obtained for at least two transcriptional units in the rfb gene cluster and the approximate locations of two promoters are suggested . The detection of new electrophoretic species of LPS that may correspond to LPS biosynthetic intermediates, and the finding on the cell surfaces of structures corresponding to LPS core substituted with one or more O-specific sugars, appear to be novel findings. Microb Pathog, 1986 Jun, 1(3), 299 - 306 Genetic and biochemical analysis of Shigella dysenteriae 1 O antigen polysaccharide biosynthesis in Escherichia coli K-12: 9 kb plasmid of S . dysenteriae 1 determines addition of a galactose residue to the lipopolysaccharide core; Sturm S et al.; Production of the somatic antigen, O-specific polysaccharide of Shigella dysenteriae 1 is determined by the chromosomal rfb gene cluster and the rfp gene located on the 9 kb plasmid pHW400 carried by this organism . When transferred to Escherichia coli K-12, which produces lipopolysaccharide consisting only of core oligosaccharide linked to lipid A, rfp gene-containing plasmids caused modification of the core oligosaccharide leading to the appearance of core molecules with new electrophoretic mobilities . Chemical analysis of the modified core has shown that it is substituted with a galactose residue which is the first sugar of the O-polysaccharide repeat unit. Microb Pathog, 1986 Jun, 1(3), 289 - 97 Cloning of the rfb gene region of Shigella dysenteriae 1 and construction of an rfb-rfp gene cassette for the development of lipopolysaccharide-based live anti-dysentery vaccines; Sturm S et al.; Recent studies have shown that determinants for the production of O antigen lipopolysaccharide in Shigella dysenteriae 1 are distributed over two distinct genetic elements, the chromosome and a 9 kb plasmid designated pHW400 . In this communication, we describe the cloning of all determinants necessary for S . dysenteriae 1 O antigen production in E . coli K-12 and their combination in a single plasmid . An RP4::miniMu R-prime plasmid, R-prime 40, containing the his-rfb (histidine biosynthesis-lipopolysaccharide biosynthesis) gene region of the Shigella dysenteriae 1 chromosome was generated . E . coli K-12 bacteria containing R-prime 40 and pSS8, a transposon Tn5-tagged derivative of pHW400, produced lipopolysaccharide indistinguishable from that of S . dysenteriae 1 . Small DNA fragments containing the rfb gene cluster and the rfp gene were subcloned from R-prime 40 and pSS8 and subsequently combined in vector pACYC184 to produce pSS37 . This latter plasmid when introduced by transformation into E . coli K-12 provoked the formation of S . dysenteriae 1 O-specific lipopolysaccharide, a feature that suggests it may be useful in the construction of LPS-based live vaccines against the Shiga bacillus. Antimicrob Agents Chemother, 1986 May, 29(5), 883 - 6 Comparison of norfloxacin and nalidixic acid for treatment of dysentery caused by Shigella dysenteriae type 1 in adults; Rogerie F et al.; A severe epidemic of dysentery began late in 1979 in northeast Zaire and spread to Rwanda, Burundi, and Tanzania . The epidemic strain is a multiply resistant Shigella dysenteriae type 1, which acquired resistance against trimethoprim and more recently against nalidixic acid in the course of the epidemic . A comparative open trial in Rwandan adults with Shiga dysentery involved 18 patients treated with norfloxacin at 400 mg twice daily and 12 patients treated with nalidixic acid at 1 g three times daily for 5 days . All isolates showed in vitro susceptibility to both drugs . Though norfloxacin eliminated Shigella organisms from stools more rapidly than nalidixic acid, its clinical superiority did not reach the level of significance . Norfloxacin is a promising drug and is more effective than nalidixic acid in the treatment of multiresistant shigellosis. Zh Mikrobiol Epidemiol Immunobiol, 1986 May, (5), 29 - 32 {Characteristics of the tissue population cycle of shigellae}; Bondarenko VM et al.; In experiments on human fetal intestinal explants infected with shigellae the specific multiplication rate of these infective agents, found to be 0.026, and the maximum level of their accumulation in erythrocytes, reaching 22-36 microbes per cell, have been determined . These phenomena can be observed after at least 3-hour incubation and end in the release of the infective agents from the affected area with shedding epithelial elements (villi) . Shigellae, aggregated in the shed villi easily adhering to the unaffected mucosa, ensure the intensive invasion of the epithelium, which leads to the continuation of the process . The regularity thus revealed indicates that the population cycle of the development of shigellae is limited by short intervals of 3-4 hours . During these intervals the repeated invasion and the release of shigellae, together with the shed epithelium, into the chyme-containing intestinal cavity occur . The conditions for the multiplication of shigellae and their specific multiplication rate in chyme are minimal (0.016). Zh Mikrobiol Epidemiol Immunobiol, 1986 May, (5), 25 - 8 {Biochemical properties of Shigella flexneri and their practical significance}; Priamukhina NS et al.; As the result of the study of 921 S . flexneri strains 1-6 and 4 (IV: 7,8), isolated in 31 regions of the USSR in 1975-1984, their biochemical characterization by 33 tests was made . All the strains under study proved to be typical in most of their constant signs, only some of strains 2a showed deviations in mannitol and some of strains 4a, in acetate . In strains of serovar 6, circulating in the USSR, specific features with respect to dulcitol and xylose were noted . The possibility of the biochemical subserovar typing of S . flexneri 1-5, X- and Y-var., with respect to maltose, arabinose, sorbitol and rhamnose was confirmed. Rev Infect Dis, 1986 May-Jun, 8 Suppl 2, S217 - 22 Antimicrobial therapy for travelers' diarrhea; DuPont HL et al.; Three clinical trials were carried out to examine the efficacy of various antimicrobial agents in the treatment of travelers' diarrhea among students from the United States in Mexico . Thirty-seven subjects received twice daily for five days 160 mg of trimethoprim (TMP) and 800 mg of sulfamethoxazole (SMZ), 38 received 200 mg of TMP, and 35 received a placebo . Another group of students were given 100 mg of furazolidone (47 students) or ampicillin (47 students) four times a day for five days . In the third study, 500 mg of bicozamycin (72 students) or a placebo (68 students) was given four times a day for three days . Most students who received TMP-SMZ (78%), TMP (84%), or bicozamycin (85%) had recovered by 48 hr after initiation of treatment, as compared with 14% and 47% in the corresponding placebo groups and 55% in the furazolidone group . The agents had a positive effect for all etiologic categories, including diarrhea due to enterotoxigenic Escherichia coli and Shigella strains and illness without any established etiologic agents . Treatment failures were unusual with TMP-SMZ, TMP, and bicozamycin therapy (5% vs . 39% for the placebo-treated students) . The drugs were well tolerated . The use of TMP-SMZ or TMP alone in the empiric treatment of moderate to severe travelers' diarrhea is advocated. Rev Infect Dis, 1986 May-Jun, 8 Suppl 2, S167 - 71 Antimicrobial agents in the prevention of travelers' diarrhea; DuPont HL et al.; Each of 433 adults traveling to Guadalajara, Mexico, from the United States during summer months was enrolled in one of four clinical trials of the protective effect of antimicrobial agents against travelers' diarrhea . Only one (2%) of 57 subjects taking trimethoprim-sulfamethoxazole (160 mg/800 mg daily) experienced diarrhea during a two-week study, whereas eight (14%) of 58 subjects taking trimethoprim alone (200 mg daily) and 10 (33%) of 30 taking placebo developed illness (P less than .05 and P less than .0001, respectively) . Diarrhea occurred significantly less frequently among subjects receiving trimethoprim than among placebo recipients (P less than .05) . None of 11 students given bicozamycin (500 mg four times daily) developed diarrhea during a three-week study, whereas 10 (53%) of 19 placebo recipients became ill (P = .003) . Four (7%) of 54 subjects receiving norfloxacin (400 mg daily) experienced diarrhea during a two-week study; in contrast, 34 (60%) of 57 placebo recipients developed diarrhea (P less than .0001) . The various antimicrobial agents prevented illness due to enterotoxigenic Escherichia coli and Shigella as well as that unassociated with a pathogen . The drugs were well tolerated . Current evidence suggests that trimethoprim-sulfamethoxazole is the optimal antimicrobial agent available for prophylaxis of travelers' diarrhea. Ann Inst Pasteur Microbiol, 1986 May-Jun, 137A(3), 267 - 78 Plasmid-mediated contact haemolytic activity in Shigella species: correlation with penetration into HeLa cells; Clerc P et al.; The main feature of virulent strains of Shigella is their ability to invade eucaryotic cells . This phenotype is both plasmid-mediated and temperature-regulated . In the present report, we demonstrate a plasmid-mediated and temperature-regulated haemolytic activity in S . flexneri, S . dysenteriae and S . sonnei . Detection of this haemolytic activity requires centrifugation of suspensions containing bacteria and erythrocytes, followed by incubation of the pellets at 37 degrees C . No soluble intra- or extracellular haemolytic activity could be detected . Dose-effect and electron microscopic studies demonstrated that direct contact of several virulent bacteria per erythrocyte was critical for haemolysis to occur . However, no local morphological alteration of the erythrocyte membrane at the site of contact with bacteria could be detected . Intensity of this haemolytic activity was fully correlated with the efficiency of penetration within HeLa cells, suggesting a common mechanism for induction of phagocytosis and lysis of erythrocytes. J Bacteriol, 1986 May, 166(2), 375 - 9 Cloning and expression of the genes specifying Shiga-like toxin production in Escherichia coli H19; Huang A et al.; Some strains of Escherichia coli produce a protein which is cytotoxic for Vero cell and HeLa cell monolayers . This toxin is very similar to the toxin of Shigella dysenteriae 1 and has been named verotoxin or E . coli Shiga-like toxin . It has been shown that toxin conversion is due to a group of bacteriophages, one of which has been designated H-19B . In this study we report hybridization experiments showing that part of the H-19B genome is homologous to phage lambda . We have cloned a 1.7-kilobase BalI-BglII fragment from the genome of H-19B into pUC18 . The recombinant plasmid confers the ability to produce high levels of Shiga-like toxin on transformed E . coli cells . We demonstrate using an in vitro transcription/translation system that the cloned fragment specifies the two verotoxin subunit peptides which have masses of 31 and 5.5 kilodaltons . The identity of peptides was confirmed by immunoprecipitation with verotoxin antiserum and protein A-Sepharose beads. Am J Epidemiol, 1986 Apr, 123(4), 710 - 20 Breast feeding as a determinant of severity in shigellosis . Evidence for protection throughout the first three years of life in Bangladeshi children; Clemens JD et al.; PIP: The influence of breastfeeding on the severity of illness in shigellosis was assessed in a case-control study involving 540 children presenting with shigellosis to a Bangladesh diarrheal disease hospital in 1980-82 . The 53 cases were children under 3 years of age with severe illness (i.e., rectal temperature above 102 F, severe neurologic manifestations, or severe dehydration), while the 487 age-matched controls had non-severe shigellosis . 42% of the cases compared with 59% of controls were breastfed, yielding an odds ratio of 0.49 (95% confidence interval, 0.28-0.86) . This finding suggests that breastfeeding substantially shifts the spectrum of severity in Shigella infections from severe to non-severe illness . The high degree of protection against severe shigellosis was also seen in malnourished children and in children reporting a recent history of measles . The protective effect of breastfeeding persisted even when the analysis was corrected for the confounding effects of age, nutritional status, and earlier receipt of medication . These results are consistent with data from both developed and developing countries indicating that breastfeeding reduces the severity of illness in childhood diarrhea . In Bangladesh, about 1/3 of mothers breastfeed for 3 years . In other areas of the developing world, however, breastfeeding beyond infancy is relatively uncommon . Because shigellosis accounts for substantial morbidity and mortality in developing countries, prolonged breastfeeding is recommended . Teratology, 1986 Apr, 33(2), 243 - 6 Effect of Shigella toxin on preimplantation mouse embryos in vitro; Olsen WM et al.; Preimplantation mouse embryos at the 4-cell to 8-cell stage were exposed to Shigella dysenteriae toxin at concentrations of 0.001-100 pg/ml in vitro . The effect of the toxin was studied by morphological observation of the embryos to the blastocyst stage, by assessing protein synthesis with 14C-leucine incorporation, and by measuring embryonic adenosine triphosphate (ATP) content . Preimplantation mouse embryos were highly sensitive to the toxin . All variables investigated were adversely influenced by the toxin . After a lag period of 24 hr, 0.01 pg/ml toxin inhibited development to the blastocyst stage and protein synthesis . Toxin concentrations of 1.0 pg/ml resulted in a significant decrease in ATP content. Zh Mikrobiol Epidemiol Immunobiol, 1986 Apr, (4), 21 - 4 {Dynamics of Shigella interaction with the epithelium in the infection process}; Zhalko-Titarenko VP et al.; The study of the adhesion of Shigella flexneri to intestinal mucosal explants from human postabortion fetuses, used as an experimental model, has revealed that the process of interaction between the infective agent and the epithelium develops in accordance with Langmuir's equation of the adsorption isotherm . The specific biological feature of the adhesive interaction between bacteria and the mucous membrane is the fact that the effective adhesion of microbes is possible only in case of their high concentration on the surface of the mucous membrane . In case of their low concentration in the parietal layer no microbial adhesion is observed, whereas epithelial villi infected with shigellae and fixed to the mucous membrane of the explant produce a high parietal concentration of the infective agents, which leads to the increase of adhesion by more than two orders. Antimicrob Agents Chemother, 1986 Apr, 29(4), 645 - 8 Comparative efficacies of single intravenous doses of ceftriaxone and ampicillin for shigellosis in a placebo-controlled trial; Kabir I et al.; To evaluate ceftriaxone for the treatment of shigellosis, 94 adult males with acute dysentery were randomly assigned to receive ceftriaxone (1 g), ampicillin (4 g), or saline placebo intravenously in single doses in a double-blind design . Stool cultures were positive for Shigella dysenteriae in 52 patients, S . flexneri in 38 patients, and other species in 4 patients . Both ceftriaxone and ampicillin caused reductions in the mean duration of fever and the means of daily stool frequency 2 to 4 days after therapy versus placebo (P less than 0.05) . The ability of ceftriaxone to reduce stool frequency during 6 days after treatment was significant in patients with S . flexneri infections (P less than 0.05), whereas S . dysenteriae infections were relatively refractory to improvement by both antibiotics . Neither drug had a significant effect on overall duration of diarrhea, blood in stool, or tenesmus . Ampicillin reduced the mean duration of positive stool cultures after treatment from 2.6 days in the placebo group to 1.1 days (P less than 0.05), whereas ceftriaxone did not affect the duration of Shigella sp . excretion . These results indicate that single intravenous doses of ceftriaxone and ampicillin caused some clinical improvement in acute shigellosis but only ampicillin exerted a bacteriological effect on Shigella sp . excretion. J Clin Microbiol, 1986 Apr, 23(4), 715 - 7 Feasibility of same-day identification of members of the family Vibrionaceae by the API 20E system; Overman TL et al.; Sixty isolates, comprising nine species of the family Vibrionaceae, were tested with the API 20E 5-h same-day procedure (Analytab Products, Plainview, N.Y.) . Included were 27 Aeromonas hydrophila isolates, 10 Aeromonas sobria isolates, 7 Aeromonas caviae isolates, 3 Plesiomonas shigelloides isolates, 3 Vibrio alginolyticus isolates, 3 Vibrio cholerae isolates, 1 Vibrio fluvialis isolate, 5 Vibrio parahaemolyticus isolates, and 1 Vibrio vulnificus isolate . The 5-h profile numbers were specific for the five Vibrio species and the Plesiomonas isolates . The three Aeromonas species shared seven 5-h profile numbers . Of the 63 5-h profile numbers generated by testing each isolate twice, 22 were identical to those found in the overnight analytical profile index . Of these, 20 were correct identifications, and two were incorrect . The remaining 41 5-h profile numbers were not found in the overnight analytical profile index . Because the 5-h analytical profile index does not contain any oxidase-positive organisms, the oxidase value was subtracted from the 63 5-h profile numbers to determine whether misidentifications could occur if the oxidase test was either not performed or not performed correctly . Only five of these factored profile numbers resulted in a possible misidentification . It is feasible, within limitations, to use the 5-h API 20E same-day procedure to identify the more commonly occurring members of the Vibrionaceae . The manufacturer should develop a data base for this purpose. Am J Pathol, 1986 Apr, 123(1), 25 - 38 Ultrastructural pathology of the rectal mucosa in Shigella dysentery; Mathan MM et al.; Colonocyte damage in the rectal mucosa in shigellosis is the result of bacterial invasion and leads to ulceration . Additional factors in colonocyte damage may be the Shigella cytotoxin and, especially in colonic crypt cells, bacterial endotoxin . A vascular lesion was present in the lamina propria of the rectal mucosa, which resembled endothelial damage secondary to bacterial endotoxins . In patients with longer duration of symptoms, relative vascular insufficiency, activated lymphocytes, eosinophil and mast cell degranulation, and antibody-mediated colonocyte damage may all play a role. Zh Mikrobiol Epidemiol Immunobiol, 1986 Apr, (4), 40 - 4 {Risk factors in the epidemiology of dysentery}; Zamotin BA et al.; The general tendency of a decrease in the morbidity rates of dysentery induced by Shigella sonnei and Shigella flexneri (separately) from Monday to the end of the week (Saturday-Sunday) has been revealed, and at the same time the "infection risk" for both kinds of salmonellosis has been found to fall on the last days of the week (Friday-Sunday). Int J Epidemiol, 1986 Mar, 15(1), 116 - 21 Risk factors for diarrhoeal deaths: a case-control study at a diarrhoeal disease hospital in Bangladesh; Islam SS et al.; The study of diarrhoeal mortality risk has been limited to descriptive epidemiological investigations . This is the first case-control study in this area, and our results show that certain risk factors are predictive of diarrhoeal deaths . We have compared the disease history, associated complications, signs, symptoms and laboratory values of 346 patients who died on the medical wards of Dhaka Hospital and that for 346 matched controls to identify the risk factors for mortality . Patients presenting with oedema, severe dehydration or convulsion were found to have a risk of mortality two times higher than controls . Several laboratory results were compared by a matched pair analysis, demonstrating that hyponatraemia (less than 130 mmol/L), hypobicarbonaemia (less than 20 mmol/L) and raised anion gap (greater than 14.9 mmol/L) were moderately associated with mortality . However, hypoproteinaemia (less than 50 gm/L) was strongly associated . For children less than 10 years of age, both hyper- and hyponatraemia were found to be associated with mortality, and the nutritional status of the children modified the effect of hypernatraemia on diarrhoeal mortality . The strong association between diarrhoeal death and hypoproteinaemia may be due to the effect of the pre-existing malnutrition of these patients and/or their loss of protein during shigella infection . The various risk factors that we have identified could be used as a prognostic guide by physicians treating such patients. Eur J Epidemiol, 1986 Mar, 2(1), 26 - 30 Epidemic Reiter's syndrome following an outbreak of shigellosis; Finch M et al.; We prospectively studied the occurrence of Reiter's syndrome (RS) or reactive arthritis (ReA) in 205 of 349 cruise-ship passengers who attended a buffet ashore and developed Shigella flexneri 2a enteritis . Five passengers probably had RS/ReA and 16 were possible or doubtful cases of RS/ReA . HLA-B27 was identified in 4 of 5 probable RS cases, but was not present in any of the 16 possible or doubtful cases nor in any of 20 passengers (controls) without any symptoms of RS/ReA . There was no statistically significant difference in the frequency of B7-Creg antigens in persons with possible or doubtful RS/ReA (9/16) compared to controls (8/20). Cent Afr J Med, 1986 Mar, 32(3), 79 - 82 An epidemic of bacillary dysentery in western Rwanda 1981-1982; Huppertz HI; PIP: The epidemiologic and clinical aspects of a major Shigella dysenteriae epidemic resistant to all available drugs that occurred in Western Rwanda in 1981-82 are analyzed . About 5% of the population was affected . In total, there were 11,801 cases of dysentery, with an average of 833 cases/month . The course of the epidemic was independent from seasonal variations . In children under 5 years of age, the majority (61%) of dysentery cases involved males; at all other ages, however, there was a female preponderance . The duration of illness averaged 1 week . Since no effective chemotherapy could be found, treatment consisted of general supportive measures and rehydration . The overall case fatality rate 6%, with higher rates among children (12%) and adult men (9%) . Dehydration and fever were the 2 signs most frequently associated with a fatal outcome . 33 women followed in this study were pregnant while suffering from dysentery . 5 abortions, 3 premature deliveries, and 1 intrauterine death were recorded, suggesting that epidemic dysentery poses a serious risk to pregnant women . Disease transmission was primarily through direct contact . Health education is the most effective way to improve hygienic conditions among the peasant population of Rwanda and to decrease the risk of infection . Antibiot Med Biotekhnol, 1986 Mar, 31(3), 215 - 7 {Conjugative R-plasmids simultaneously belonging to the 2 incompatibility groups INC I zeta and INC B}; Shishkina ZV et al.; Five plasmids detected in the clinical strains of Shigella were studied . The plasmids had similar phenotypes of antibiotic resistance and allotted streptomycin and tetracycline resistance to the host cell . None of them allotted the bacterial cells with sensitivity to phages specific for the Inc groups F, N, P and I . The molecular weights of the plasmids ranged from 47 to 50 MD . The studies on the incompatibility showed that pKMR 227, pKMR 228-1, pKMR 231, pKMR 242-1 and pKMR 245 plasmids simultaneously belonged to two Inc groups: Inc I zeta and Inc B . Two plasmids, i.e . R 805a and R 861a simultaneously belonging to the two Inc groups: Inc I zeta and Inc B are described in the literature . pKMR 231 and pKMR 242-1 plasmids detected by the authors differ from them by the antibiotic resistance phenotype . The other three pKMR plasmids in addition differ from them by belonging to Fin. Zh Mikrobiol Epidemiol Immunobiol, 1986 Mar, (3), 75 - 8 {Bactericidal and immunocorrective properties of plant extracts}; Kolesnikova AG; Extract from oak cork, St . John's wort leaves and flowers and pine buds possess more pronounced bactericidal properties with respect to staphylococci, shigellae, Escherichia coli than decoctions from these medicinal plants . Such extracts may be included into complexes for the treatment of otorhinolaryngological diseases, enterocolitis in children and bacterial eczema . As a medicinal form, extract prepared from medicinal plants are more convenient for storage and permit more exact dosage. Plasmid, 1986 Mar, 15(2), 119 - 31 Characterization of transferable plasmids from Shigella flexneri 2a that confer resistance to trimethoprim, streptomycin, and sulfonamides; Chinault AC et al.; A set of plasmids conferring resistance to several antibiotics, including the combination of trimethoprim and sulfamethoxazole, has been isolated from Escherichia coli following conjugative cotransfer from a clinical isolate of Shigella flexneri 2a . One of the plasmids, pCN1, was shown by subcloning and DNA sequencing to carry a gene encoding a trimethoprim-insensitive dihydrofolate reductase identical to that found in E . coli transposon 7 . This plasmid was also shown to confer resistance to both streptomycin and spectinomycin by production of an adenylyltransferase that inactivated the drugs and the gene encoding this enzyme has also been sequenced . A second plasmid from the set, pCN2, was shown to inactivate streptomycin by a phosphotransferase mechanism and also to confer resistance to sulfonamides . The third plasmid from the set could not be correlated with a drug-resistance phenotype, but does appear to play a crucial role in plasmid mobilization. J Med Microbiol, 1986 Mar, 21(2), 117 - 23 Adherence of non-fimbriate entero-invasive Escherichia coli O124 to guinea pig intestinal tract in vitro and in vivo; Ashkenazi S; The adherence properties of non-fimbriate entero-invasive Escherichia coli O124 in the guinea pig intestinal tract were studied . Quantitative in-vitro determinations were done by incubating radiolabelled bacteria with suspensions of viable intestinal cells released by treating loops of the guinea pig intestine with solutions containing EDTA, dithiothreitol and citrate . Non-bound bacteria were separated from the intestinal cells on a Percoll gradient . Only cells released from the colon, especially from its transverse and descending regions, avidly adhered to E . coli O124 (68-79 bacteria/cell), whereas the attachment to ileal cells was negligible . The adherence process was Ca++ and temperature-dependent, had an optimal pH of 6.2 and was inhibited by fucose, glucose or mannose . Several pretreatment studies of the bacteria or the colonic cells showed that the adherence was mediated by a carbohydrate-binding protein (adhesin or lectin) on the colonic cells and not on the bacterial surface . Results of studies of in-vitro adherence to intestinal loops and to intact intestinal surfaces correlated well with the in-vitro findings . These results indicate that the adherence of entero-invasive E . coli O124 to the gut is similar to the attachment of Shigella flexneri and is quite different from that of enterotoxigenic E . coli. Gastroenterology, 1986 Mar, 90(3), 654 - 60 Rectal histology in acute bacillary dysentery; Anand BS et al.; A recent epidemic of acute Shigella dysentery in West Bengal (India) provided us with an opportunity to examine the rectal mucosal abnormalities seen in this condition . One hundred two patients were investigated using sigmoidoscopy, rectal biopsy, and rectal swab for culture . Pure culture of Shigella was obtained in 37 cases, and the rectal biopsy specimens from these patients were assessed in detail . The mean (+/- SD) duration of illness was 47.8 +/- 27.4 h (range 8-120 h), and most patients (31 of 37, 84%) had diarrhea with blood and mucus in the stools . Significant findings at histology were as follows . (a) Cellular infiltrate was predominantly round cell or mixed round cell and neutrophilic in the majority of patients (27, 73%) . (b) Disorganization of crypts was seen in as many as 31 patients (84%); in most subjects the distorted architecture was mild, but in a few the defect was severe with crypt branching and dilatation . (c) In the majority of patients the inflammatory process extended to the muscularis mucosae and submucosa; edema with or without increased cellular infiltrate was seen in the muscularis mucosae in 92% and in the submucosa in 80% . (d) There was no difference in the rectal histology of patients with a short history of disease (less than 48 h) compared with those with a longer history, except for goblet cell depletion which was more in those with diarrhea for more than 48 h . (e) The mucosal abnormalities in patients with watery diarrhea were, in general, milder than in those with dysentery, although the difference was statistically not significant; 2 of 6 patients with watery diarrhea had severe colitis . (f) The mucosal abnormalities were more severe in patients with Shigella dysenteriae infection compared with Shigella flexneri. Isr J Med Sci, 1986 Mar-Apr, 22(3-4), 277 - 82 Cost-benefit of mass prophylaxis with immune serum globulin to control waterborne hepatitis A: a case study; Egoz N; Following a point-source epidemic of more than 8,000 cases of Shigella sonnei, which was caused by a sewage-contaminated water supply, immune serum globulin (ISG) was administered to 11,306 children, aged 5 to 9 years, and to 1,338 pregnant women to prevent a possible outbreak of hepatitis A . A cost-benefit analysis showed that the estimated cost of preventing one possible case of hepatitis in a child was $362.50, and in a pregnant women $11,514 . The benefit:cost ratios were 0.45 and 0.28, respectively . These relatively low ratios call for reconsideration of the guidelines that recommend ISG administration to children and pregnant women following an exposure to contaminated water. Infect Immun, 1986 Mar, 51(3), 975 - 8 Effect of guinea pig or monkey colonic mucus on Shigella aggregation and invasion of HeLa cells by Shigella flexneri 1b and 2a; Dinari G et al.; The effects of guinea pig and rhesus monkey colonic mucus preparations on Shigella aggregation and invasion of HeLa cell monolayers by Shigella flexneri serotype 1b, 2a, and 5 strains were investigated . Guinea pig mucus caused agglutination of S . flexneri serotype 1b but not of S . flexneri serotype 2a or 5 . Guinea pig mucus also inhibited HeLa cell invasion by S . flexneri serotypes 1b and 2a . Monkey mucus neither agglutinated any Shigella strain nor inhibited HeLa cell invasion. JAMA, 1986 Feb 14, 255(6), 757 - 60 Comparison of loperamide with bismuth subsalicylate for the treatment of acute travelers' diarrhea; Johnson PC et al.; Loperamide hydrochloride was compared with bismuth subsalicylate for the treatment of acute nondysenteric travelers' diarrhea in 219 students visiting seven countries in Latin America . Subjects whose condition was not improved with therapy could elect to take trimethoprim-sulfamethoxazole . Persons receiving loperamide passed fewer unformed stools when compared with the bismuth subsalicylate group during the first four hours of therapy, from four to 24 hours, and from 24 to 48 hours after therapy was initiated . Among subjects with disease due to enterotoxigenic Escherichia coli, Shigella sp, other pathogens, and unknown agents, fewer unformed stools were passed by the loperamide-treated subjects than the bismuth subsalicylate-treated subjects for all time periods studied . No significant prolongation of disease was seen in subjects with shigellosis treated with loperamide . Eight of the loperamide-treated subjects experienced constipation compared with one in the bismuth subsalicylate-treated group; otherwise, there was no difference in minor side effects experienced between both treatment groups . We conclude that loperamide is a safe and effective alternative to bismuth subsalicylate for the treatment of nondysenteric travelers' diarrhea. Infect Immun, 1986 Feb, 51(2), 696 - 8 The use of mice in the Sereny test as a virulence assay of shigellae and enteroinvasive Escherichia coli; Murayama SY et al.; We examined the possibility that mice could be used in the Sereny test instead of guinea pigs or rabbits . Although the reactions in mice were more transient and not as pronounced as those in guinea pigs, mice indeed could be used to distinguish even macroscopically between virulent and avirulent shigellae . Virulent enteroinvasive Escherichia coli strains were also positive for the mouse Sereny test . We described the macroscopic and microscopic appearance of the mouse eyes . Thus, mice are recommended for use in the Sereny test, particularly when a large number of samples are to be tested. Infect Immun, 1986 Feb, 51(2), 476 - 82 Molecular cloning of a genetic determinant for Congo red binding ability which is essential for the virulence of Shigella flexneri; Sakai T et al.; A DNA sequence of about 1.0 kilobase (kb) derived from a 230-kb (140-megadalton) plasmid in a fully virulent Shigella flexneri 2a strain, YSH6000, was cloned into Escherichia coli K-12 by selecting for the ability to bind Congo red (Pcr+ phenotype) . It was mapped and localized within the SalI restriction fragment F on the plasmid . This clone converted an S . flexneri strain which is avirulent as a result of a small deletion in the plasmid to full virulence but did not convert those without the 230-kb plasmid or with a plasmid bearing a more extensive deletion . This indicates that there are more than two virulence determinants on the plasmid . Thus, this sequence contains a genetic determinant which is essential but not sufficient for full virulence and expression of the Pcr+ phenotype in S . flexneri but is essential and sufficient for expression of the Pcr+ phenotype in E . coli K-12 . We noted that there exist some other regions on the 230-kb plasmid which express the Pcr+ phenotype in E . coli K-12 . Although these regions express the Pcr+ phenotype less markedly than the region cloned in the present study, they do hybridize with it. Infect Immun, 1986 Feb, 51(2), 461 - 9 Multiplication of Shigella flexneri within HeLa cells: lysis of the phagocytic vacuole and plasmid-mediated contact hemolysis; Sansonetti PJ et al.; Virulent strains of Shigella flexneri invade HeLa cells with high efficiency . This crucial step in the pathogenic process is encoded by a 140-megadalton plasmid which induces phagocytosis of the bacteria by host cells . In this report we used pWR100, the virulence plasmid of S . flexneri serotype 5, and pHS4108, a 32-megadalton subclone of pWR100, to demonstrate that the plasmid is also responsible for rapid intracellular growth of the bacteria . The ability to replicate intracellularly was not correlated with induction of Shiga toxin . However, plasmid-mediated intracellular multiplication was strongly correlated with the ability of the bacteria to rapidly and efficiently lyse the phagocytic vacuole and replicate freely in the cytoplasm . Temperature-regulated plasmid-mediated contact hemolytic activity strongly correlated with both phagosomal membrane lysis and efficient intracellular multiplication . We propose this virulence plasmid-associated hemolysin to be an important factor in the invasion and proliferation of Shigella spp . in mammalian cells. J Immunol, 1986 Feb 1, 136(3), 1081 - 6 Natural cytotoxic effector cell activity against Shigella flexneri-infected HeLa cells; Klimpel GR et al.; Virus and facultative intracellular bacteria both replicate within a host cell . The recognition and killing of virus-infected cells by natural killer (NK) cells is thought to be an important host immune function . However, little is known about immune recognition of bacteria-infected cells . In this report, we show for the first time that human peripheral blood lymphocytes (PBL) and large granular lymphocytes (LGL) purified from PBL have significant levels of cytotoxic activity against Shigella flexneri-infected HeLa cells . This cytotoxic activity was dependent on bacterial invasion of the HeLa cells, because HeLa cells pretreated with a noninvasive isogenic variant of S . flexneri or soluble bacterial products were not killed . Pretreatment of PBL with interleukin 2 (IL 2) or interferon-alpha greatly enhanced the cytotoxic activity of PBL against Shigella-infected HeLa cells . Cytotoxic activity present in PBL or in PBL pretreated with IL 2 was shown to be associated with both Leu-11+ and Leu-11- cell populations . These results suggest that NK cell killing of bacteria-infected cells may play an important role in host defense against facultative intracellular bacterial infections. J Infect Dis, 1986 Feb, 153(2), 238 - 48 Pathogenesis of shigella diarrhea: evidence for an N-linked glycoprotein shigella toxin receptor and receptor modulation by beta-galactosidase; Keusch GT et al.; Pathogenic mechanisms in infectious diseases often involve specific receptor-ligand interactions of cells and soluble molecules . To further elucidate structure-function relations for shigella toxin receptors, we studied binding of purified 125I-labeled toxin and biologic response under various conditions in an experimental model using HeLa cells . Response to toxin was reversibly inhibited by treatment of cells with trypsin or tunicamycin, an inhibitor of glycoprotein synthesis that also significantly inhibited toxin binding, a result indicating that the receptor is an N-linked glycoprotein . Removal of terminal beta-linked galactose from the HeLa cell surface with beta-galactosidase increased toxin binding and activity, and it also potentiated the effects of lysozyme and wheat-germ agglutinin, which recognize oligomeric beta 1----4-linked N-acetyl-D-glucosamine and inhibit toxin activity as well . Incubation of cells with beta-N-acetylglucosaminidase, which cleaves terminal beta-linked N-acetyl-D-glucosamine, inhibited toxin activity . Effects of beta-galactosidase were reversed by readdition of galactose to cell-surface oligosaccharide acceptors . The data demonstrate that alterations of a single sugar on cell-surface glycoproteins may have a dramatic effect on receptor activity and indicate that shigella toxin is a sugar-binding protein with specificity for beta 1----4-linked N-acetyl-D-glucosamine. Infect Immun, 1986 Feb, 51(2), 470 - 5 Molecular alteration of the 140-megadalton plasmid associated with loss of virulence and Congo red binding activity in Shigella flexneri; Sasakawa C et al.; A plasmid of about 140 megadaltons has been associated with the invasiveness of Shigella flexneri . Upon subculturing in liquid media of fully virulent isolates of Shigella flexneri 2a YSH6000, which contains only a 230-kilobase-pair (kbp) plasmid in addition to 3.3- and 4.2-kbp cryptic plasmids characteristic to all S . flexneri strains, loss of invasiveness, loss of Congo red binding activity (Pcr), and complete loss of, or a deletion, or even a single-site IS insertion in the plasmid occurred simultaneously . This was ascribed to the fact that, once a noninvasive Pcr- cell has emerged, it overgrows the wild type as a consequence of its selective advantage in artificial media . A deletion map of the 230-kbp plasmid was made by analyzing SalI digests of 39 deletion derivatives plus 1 formed by insertion of an IS1-like element in independently isolated, noninvasive Pcr- mutants . Of 39 deletion derivatives, 16 belonged to a single type, and 6 belonged to another, suggesting deletion hot spots . The deletion map was confirmed and extended by analyzing 359 SalI-generated partial digests of the wild-type plasmid cloned into pBR322 . Three copies of IS1-like elements were found on three different SalI fragments by Southern hybridization . Segments required for the Pcr+ phenotype seemed to occur at several different locations in the plasmid . Each of 28 representative Pcr- mutants were negative by the Sereny test . Hence, many, or possibly all, Pcr determinants were required for full virulence. Bull Soc Pathol Exot Filiales, 1986, 79(4), 435 - 46 {Bacteriological study of shigellosis in the Lake Kivu area (Central Africa) . Developments in the last 15 years (1968-1983)}; Rogerie F et al.; 1913 Shigella strains were isolated from 1968 to 1983 in lake Kivu area (Central Africa) . Distribution and evolution of serotypes, biotypes and antibiotypes were studied . The majority of serotypes and biotypes were stable . However, few of these strains showed variations, among them Shigella dysenteriae serotype 1, causing epidemic dysenteriae since 1981 . Epidemiologic survey of shigellosis in Kivu seems to be rather underdeveloped (4 bacteriology laboratories for an area of 10 million inhabitants) and the number of registered cases is probably underestimated . This problem is threatening the public health and coordination of laboratory activities, either local or regional, should be improved. J Basic Microbiol, 1986, 26(5), 283 - 7 Surface hydrophobicity of plasmid-carrying virulent Shigella flexneri and their avirulent variants; Seltmann G et al.; Five pairs of strains of S . flexneri each differing in the colour of their colonies after growth on Congo red agar have been tested for their ability to cause keratoconjunctivitis in the guinea pig eye, for the presence of the 140 Md virulence plasmid, for the presence of the virulence marker antigen, and for their ability to adsorb to hydrophobic surfaces (cellulose nitrate filters and Phenyl Sepharose) . The results suggest that the presence of the 140 Md virulence plasmid provides the bacterial surface with a rather high degree of hydrophobicity; exceptions have been found. Zh Mikrobiol Epidemiol Immunobiol, 1986 Jan, (1), 58 - 62 {Population migrations and the epidemic process of Flexner and Sonne dysentery}; Kolganov AV; Analysis of the materials from the Khabarovsk Territory for the representative period of 25 years (1956-1980) revealed the influence of the migration of the population on the level and dynamics of morbidity in different kinds of dysentery, Flexner's dysentery shown to occur more frequently than Sonne dysentery . Intensive migrations increase the proportion of susceptible persons among the population, thus facilitating the formation and circulation of Shigella strains with pronounced virulence. J Int Med Res, 1986, 14(1), 19 - 29 The use of furoxone: a perspective; Phillips KF et al.; Acute infectious diarrhoea is a widespread cause of morbidity and mortality . Some of the major diarrhoeal diseases are cholera, typhoid fever, shigellosis (bacillary dysentery), salmonellosis, "travellers' diarrhoea", and giardiasis These diseases can be avoided with proper education, sanitation, and hygiene . However, the majority of these diseases occur most frequently in areas of the world where political and social upheaval, poverty, overcrowding, and a lack of education prevail . Although vaccines are available for some of the diseases, they are not completely effective . Antimicrobial therapy is effective in decreasing the duration and severity of diarrhoea and in reducing the likelihood of relapses, complications, and death . An antimicrobial drug for the treatment of acute infectious diarrhoeal disease must be relatively specific, effective, and safe, and it should not promote the development of resistant bacteria . Furazolidone (Furoxone) has been used for 30 years for the specific and symptomatic treatment of bacterial or protozoal diarrhoea and enteritis caused by susceptible organisms . Its effectiveness has often been shown to be comparable or superior to that of other drugs . In addition, the toxicity of furazolidone is relatively low, and it minimizes the development of resistant organisms . These characteristics should contribute to the continued use of furazolidone as a rational choice in the treatment of acute infectious diarrhoeal diseases that occur worldwide. Pediatr Infect Dis, 1986 Jan-Feb, 5(1 Suppl), S29 - 43 Pediatric diarrhea: the challenge of prevention; Levine MM et al.; PIP: Conditions such as poverty, underdevelopment, and lack of education facilitate the widespread transmission of the pathogens that cause diarrheal disease, dysentery, and enteric fever in young children . Such infections produce high rates of morbidity, mortality, and adverse nutritional consequences in the first 2 years of life . Although rapid socioeconomic development has produced a precipitous decline in mortality due to diarrheal disease in the developed world, such a trend is not likely in developing countries unless alternative measures are pursued . Nonspecific interventions pursued have included oral rehydration therapy to prevent and treat dehydration, promotion of breast feeding, health education to teach maternal technology, and early realimentation to diminish the nutritional consequences of infant diarrhea . In addition, there is reason to be optimistic about the future development of various immunizing agents against the major enteric pathogens . Epidemiologic data support the conclusion that prior natural infection with enterotoxigenic E . coli, Shigella, rotavirus, and V . cholerae OL confers protective immunity . Among the divergent approaches being followed in the development of vaccines against rotavirus are: 1) use of animal rotavirus as possible attenuated strains; 2) attenuating human rotaviruses by passage in tissue culture; 3) development of hybrid reassortant strains by coinfecting tissue cultures with both an animal strain well adapted to tissue culture and a human strain and then selecting a hybrid virus that possesses the human virus neutralization antigen but grows to higher titre in tissue culture; 4) evaluation of rotaviruses isolated from asymptomatic infected neonates in nursery outbreaks for their safety, infectivity, and immunogenicity in older children; 5) cloning a DNA copy of the RNA genus responsible for the neutralization antigens of rotaviruses; and 6) preparation of a synthetic peptide of the critical epitope of the neutralization antigen . Acta Microbiol Hung, 1986, 33(4), 341 - 4 ELISA detectable virulence marker antigen of enteroinvasive Escherichia coli is coded by a 140 megadalton plasmid; Pal T et al.; The ELISA reactive virulence marker antigen, characteristic of enteroinvasive Escherichia coli and virulent Shigella, expressed in an E . coli K-12 recipient after the 140 Md plasmid of an enteroinvasive E . coli strain had been introduced into it . Similarly to the epithelial cell penetration and Congo red binding capacities--known to be coded by the above plasmid--the virulence marker antigen expressed also only at 37 degrees C but not at 30 degrees C . These data suggest the plasmid coded nature of the virulence marker antigen. Arch Immunol Ther Exp (Warsz), 1986, 34(5-6), 513 - 6 Characterization of outer membrane proteins of virulent phase I Shigella sonnei strains and their avirulent phase I derivatives; Adamus G et al.; Comparison of polyacrylamide gel electrophoretic protein profiles of four isogenic sets of virulent phase I Sh . sonnei strains and their avirulent phase I cells revealed no differences in outer membrane protein composition between virulent and avirulent derivatives . However, significant qualitative and quantitative differences were found in composition of major outer membrane proteins between strains of different origin . All four strains tested contained one major protein of 33K . This protein was susceptible to proteolytic enzymes and was found to be heat modifiable . Other major proteins of 35K and 37K present in three strains and 36K present in one strain were identified as peptidoglycan associated proteins. Microbiol Immunol, 1986, 30(11), 1115 - 27 Characterization of purified Shiga toxin from Shigella dysenteriae 1; Yutsudo T et al.; Shiga toxin was purified from the culture supernatant of Shigella dysenteriae 1 by ammonium sulfate fractionation, DEAE-cellulose column chromatography and repeated chromatofocusing column chromatography . About 1.6 mg of purified Shiga toxin was obtained from 15 liters of culture with a yield of about 27% . The molecular weight of purified Shiga toxin was estimated to be 62,000 . The toxin consisted of A and B subunits with molecular weights of about 30,000 and 5,000-6,000, respectively . The isoelectric point of purified Shiga toxin was 7.0 . Purified Shiga toxin showed the following biological activities: lethal toxicity to mice when injected intraperitoneally with an LD50 of 28 ng per mouse; cytotoxicity to Vero cells, killing about 50% of the cells at 1 pg and all of the cells at 10 pg; and fluid accumulation in rabbit ileal loops at a concentration of more than 1 microgram. C R Acad Sci III, 1986, 303(13), 557 - 9 {Transmission of Vibrio cholerae . Possible role of the camel}; Dodin A et al.; Choleraic vibrio uses the moving means that man discovers to his own use . Camels (passive and active vehicles of large quantity of crude water) were experimentally infected with a Vibrio (tox . +) and an Aeromonas (tox . +) . Suckling young camels eliminate them for 6 days and grown-up camels for 25 days . This experience, associated with the discovery of Vibrio non 01 and Shigella in the caravan camel dungs, lead them to become a potential vector of spreading from one oasis to another. Gene, 1986, 44(1), 165 - 9 Production of restriction endonucleases using multicopy Hsd plasmids occurring naturally in pathogenic Escherichia coli and Shigella boydii; Mise K et al.; A convenient procedure has been devised for detection of restriction endonucleases in the Escherichia coli-Shigella group . With this procedure, two restriction endonucleases, designated Sbo 13 and Eco T22, were found and later were identified as isoschizomers of NruI and AvaIII, respectively . These endonucleases were shown to be produced from small multicopy plasmids . They were isolated from nonpathogenic E . coli into which the plasmids had been introduced by transformation, and purified from contaminating nuclease activity . The yield was high, 1,000 units/g of wet cells for Sbo 13 and 500 units/g for Eco T22 . Sbo 13 and Eco T22 should be preferable to NruI and AvaIII because of the high yield and ease in handling the producer cells. Acta Microbiol Hung, 1986, 33(1), 39 - 42 Studies on enterotoxin of Shigella dysenteriae type 1 . II . Systemic effects in rabbits of Shigella dysenteriae 1 enterotoxin; Asnani PJ et al.; The enterotoxin of Shigella dysenteriae type 1 is an acid and heat-labile protein . It induces a gut dilatory response and increases the levels of blood glucose, serum alkaline phosphatase and serum acid phosphatase in rabbits. Int J Epidemiol, 1985 Dec, 14(4), 607 - 13 Fourteen years of shigellosis in Dhaka: an epidemiological analysis; Khan MU et al.; We examined whether the proportion of Shigellae patients among diarrhoeal cases, the distribution, species, case-fatality rates and hospital visits changed over time in Dhaka . We isolated 19639 Shigella strains from 822812 diarrhoea cases treated at the International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR,B), between 1969 and 1982 . The number of cases increased from 209 (2.5%) in 1969 to 4833 (7.7%) in 1976 . Extrapolating from a 4% vigorous systematic sample of ICDDR,B hospital visits shigellosis cases and their proportion among diarrhoea cases increased to more than 9500 (12.0%) in 1981 . The prevalence of various shigellae species altered over time . For example: in 1969 Shigella flexneri predominated in 74% of all Shigella cases; in 1973 Shigella dysenteriae accounted for 56%, and in 1981 Shigella flexneri again predominated (75%) . More than 20% of all Shigella isolations were from infants: 60% in males and 40% in females . Over 7% of severe cases of Shigella infection referred from the outpatient department and admitted for treatment died . Nearly 40% of all the Shigella deaths were in infants of less than a year old while 49% were in 1-4 year old children . Increasing prevalence of shigellosis appears to be an important cause of diarrhoea in Dhaka especially among children . Areas with poor sanitation and water supply had higher prevalence . However, hospitalized cases represented a fraction of the actual problem . Resistance to antibiotics appears to be increasing and the development of new drugs and preventive methods within economic reach of less developed countries are crucial for reduction of the disease and related deaths. Zentralbl Bakteriol Mikrobiol Hyg {B}, 1985 Dec, 182(1), 58 - 63 {Spectrum of gram-negative rods in waste water from Jeddah/Saudi Arabia}; Muller HE et al.; 23 samples of effluent of the wastewater treatment plant of Jeddah/Saudi Arabia were investigated between September 1983 and November 1984 . A total of 395 strains of gram-negative fermentative bacteria were isolated and differentiated at species level . Neither vibrios, salmonellae, or shigellae were detected notwithstanding of enrichment procedures used . Furthermore, the occurrence of E . coli producing heat labile enterotoxin (LT) was studied using both Biken test and GM1-ELISA . The Biken test produced many false positive results which could not been confirmed by the more specific GM1-ELISA . All strains of E . coli investigated were negative . Therefore, the Biken test seems not to be a recommended method for detection of E . coli LT. J Antimicrob Chemother, 1985 Dec, 16(6), 691 - 8 Plasmid characterization of Shigella spp . isolated from children with shigellosis and asymptomatic excretors; Haider K et al.; Strains of Shigella dysenteriae type 1 and Sh . flexneri isolated during 1981-82 from children with shigellosis and also from asymptomatic excretors were examined for drug resistance and for the presence of plasmids . Shigella strains isolated from dysenteric children showed multiple plasmid bands . Thirty-eight percent of the strains transferred the drug resistance factor(s), either partially or fully to a recipient Escherichia coli K12 strain . Plasmids in the molecular weight range from between 44-76 Mdal were correlated with drug resistance . It was found that multi-resistant clinical isolates generally harbour a single large transmissible plasmid . Strains isolated from asymptomatic excretors demonstrated plasmid patterns different from those isolated from children with shigellosis although the bands were relatively homogeneous within each group . Both the groups showed the presence of a 140 Mdal plasmid band conferring invasiveness and such strains gave a positive Sereny test . This study thus shows that Shigella strains from asymptomatic excretors also retain invasiveness. J Infect Dis, 1985 Dec, 152(6), 1114 - 9 Changing pattern of resistant Shiga bacillus (Shigella dysenteriae type 1) and Shigella flexneri in Bangladesh; Shahid NS et al.; Shigella dysenteriae type 1 (Shiga bacillus) has made a dramatic comeback in Bangladesh after 10 years when Shigella flexneri was the dominant serogroup . Whereas S . flexneri showed little increase in resistance to the commonly used antimicrobial agents--namely, trimethoprim-sulfamethoxazole (TMP-SMX) and ampicillin-over the years, S . dysenteriae type 1 underwent rapid changes with regard to sensitivity pattern during the last two years . The first TMP-SMX-resistant strain of S . dysenteriae type 1 was detected in September 1982; however, by mid-1984 most strains were resistant while retaining sensitivity to ampicillin . During this period, the ratio of S . flexneri to S . dysenteriae type 1 narrowed from 0.15 to 1 . Such propagation of high resistance to TMP-SMX might have been due to widespread use of the drug imported into the country in large quantities . Resistance to ampicillin is increasing rapidly, particularly in S . dysenteriae type 1. Infect Immun, 1985 Dec, 50(3), 620 - 9 Identification and antigenic characterization of virulence-associated, plasmid-coded proteins of Shigella spp . and enteroinvasive Escherichia coli; Hale TL et al.; Seven plasmid-coded polypeptides, designated a through g, were identified by two-dimensional nonequilibrium pH gradient electrophoresis of radiolabeled extracts from minicells of virulent Shigella flexneri serotypes 2a and 5 and enteroinvasive Escherichia coli O143 . These polypeptides were deemed to be products of 140-megadalton (MDa) virulence-associated plasmids because they were not synthesized in minicells which were not harboring a 140-MDa plasmid or in minicells which were carrying an F lac plasmid of the same incompatibility group . Synthesis of these polypeptides was repressed in minicells incubated at 30 degrees C and in minicells isolated from a noninvasive opaque colonial variant, even though these strains harbored a 140-MDa plasmid . Enriched fractions of polypeptides b, c, and d were obtained from S . flexneri serotype 5 by preparative isoelectric focusing, and polyclonal rabbit antisera recognizing each polypeptide were raised . These antisera were able to detect cross-reacting plasmid-coded polypeptide antigens in S . flexneri serotype 3, Shigella sonnei, and enteroinvasive E . coli O143 . In addition, Western