Comparison of Second- and Third-Generation Cephalosporin as Initial Therapy for Women with Community-Onset Uncomplicated Acute Pyelonephritis

Purpose

This study examined the clinical effectiveness of parenteral cefuroxime and cefotaxime as empirical antibiotics for treating hospitalized women with uncomplicated acute pyelonephritis (APN).

Materials and Methods

This study was based on the clinical and microbiologic data of 255 hospitalized women with APN. Of these 255 women, 144 patients received cefuroxime and 111 received cefotaxime.

Results

There were no marked differences in the demographic features, clinical characteristics, and treatment duration between the populations of the cefuroxime and cefotaxime groups. The rates of defervescence showed no significant differences in the two groups at 48, 72, 96, and 120 hours. The clinical cure rates observed at the follow-up visit 4 to 14 days after the completion of antimicrobial therapy were not statistically different between the cefuroxime and cefotaxime groups [94.9% (129 of 136) versus 98.0% (100 of 102), respectively; p=0.307], and the microbiological cure rates were also not significantly different [88.3% (91 of 103) versus 95.0% (76 of 80), respectively; p=0.186]. The median hospitalization periods in the cefuroxime and cefotaxime groups were 7 (6-8) and 7 (6-8) days (p=0.157), respectively. Microbiological success rates after 72-96 hours of initial antimicrobial therapy were also not statistically different in the cefuroxime and cefotaxime groups, 89.4% (110 of 123) versus 94.9% (93 of 98; p=0.140).

Conclusion

Cefuroxime, a second-generation cephalosporin, is an appropriate antibiotic option for the initial treatment of uncomplicated APN and its efficacy does not differ from cefotaxime, a third-generation cephalosporin, in the initial parenteral therapy for community-onset APN.     

Antibiogram of E. coli serotypes isolated from children aged under five with acute diarrhea in Bahir Dar town

Background

Diarrheal disease and its complications remain a major cause of morbidity and mortality in children. The prevalence and antibiogram of E. coli as causative agents of diarrhea vary from region to region, and even within countries in the same geographical area.

Objectives

To determine the serotype and antimicrobial susceptibility of E. coli in children under-five years of age.

Methods

A cross-sectional study was conducted among 422 children with diarrhea from December 2011 to February 2012. Identification of E. coli and antimicrobial susceptibility testing were done following standard procedures.

Results

The overall isolation rate of E. coli was 48.3%. Poly 2 sero-groups, poly 3 sero-groups, poly 4 sero-groups and E. coli O157:H7 accounted for 80 (39.2%), 40 (19.6%), 25 (12.3%), and 59 (28.9%) of the isolates, respectively. Poly 2 sero-groups, constituting isolates belonging to enteropathogenic E. coli were the most commonly isolated serotypes. E. coli exhibited high levels of antimicrobial resistance to ampicillin (86.8%), tetracycline (76%) and cotrimoxazole (76%). Low levels of resistance to ciprofloxacin (6.9%) and norfloxacin (9.3%) were documented.

Conclusion

High prevalence of diarrheagenic E. coli compounded by alarming antimicrobial resistances is a serious public health problem. Regular determination of antibiogram and public education are recommended.     

A case of hemolytic uremic syndrome caused by Escherichia coli O104:H4

A 29-year-old woman presented with bloody diarrhea, abdominal pain, hemolytic anemia, thrombocytopenia, and acute renal failure. She was diagnosed with Escherichia coli O104:H4-associated hemolytic-uremic syndrome (HUS) and treated with plasmapheresis and hemodialysis for 3 weeks. She recovered without sequelae. To the best of our knowledge, this is the first report of Escherichia coli O104:H4-associated HUS in Korea. We recommend that Escherichia coli O104:H4, as well as the more common O157:H7, be considered in the diagnosis of bloody diarrhea-associated HUS.     

Effect of vitamin E nutritional supplementation on the pathological changes induced in the ileum of rabbits by experimental infection with enteropathogenic Escherichia coli

A well-established rabbit model of enteropathogenic E. coli (EPEC) disease was used to examine whether vitamin E (VE) nutritional supplementation had an effect on the pathological changes induced in the bowel by EPEC. Quantitative methods were used to evaluate the influence of VE on bacterial colonization, intestinal mucosal architecture and inflammation, and intestinal epithelial proliferation and apoptosis. VE did not affect EPEC colonization and did not give significant protection against EPEC-induced changes and diarrhoea. Although VE had no effect on the EPEC-related increase of enterocyte apoptosis, it clearly contributed to an acceleration of epithelial cell proliferation in the ileal crypts. This finding may explain why ileal morphometry undertaken in this study showed that VE ameliorated somewhat the effects of EPEC on intestinal mucosal architecture. Quantitative studies on inflammatory cells in the intestinal mucosa revealed that VE nutritional supplementation resulted in an increased neutrophilic and mononuclear inflammatory cell response to EPEC infection, which did not contribute, however, to the clearance of infection.     

Polyclonal antibodies against properly folded Dengue virus NS1 protein expressed in E. coli enable sensitive and early dengue diagnosis

The non-structural 1 (NS1) protein plays an important role in dengue diagnosis because it has been detected as a soluble serum antigen in both primary and secondary infections. The NS1 protein was expressed in Escherichia coli cells, and the efficiency of four different refolding protocols was tested. All of the protocols generated dimeric NS1 in a conformation similar to that of the protein expressed by eukaryotic cells. A polyclonal antibody produced from the properly folded E. coli recombinant NS1 (rNS1) protein proved to be a useful tool for the diagnosis of Dengue virus because it detected 100% of the Dengue virus 2 (DENV2) in infected patients’ sera and 60% of the DENV IgM-positive sera not detected by commercial NS1-based diagnostic kits. These data suggest a high-efficiency method for correctly folding rNS1 that maintains its structural and immunogenic properties. In addition, a detection method using the polyclonal antibody against correctly folded rNS1 seemed to be more sensitive and efficient for NS1 detection in serum, highlighting its usefulness for developing a high-sensitivity diagnostic kit.     

Action of bradykinin on the mesenteric microcirculation in rats

Responses of the mesenteric microvessels of rats to application and intravenous injection of bradykinin were studied in acute experiments using intravital microscopy. No clear bioelectric response of the smooth-muscle cells of the microvessels to local application of bradykinin could be found in electrophysiological experiments. In another series of experiments the dynamics of vasomotor activity were studied after the same treatment by the split-image method. The response of the microvessels to bradykinin application was shown to vary in character depending principally on whether the preparation was being applied for the first or subsequent time. It is suggested that bradykinin acts on the mesenteric microvessels through the intermediary of other vasoactive substances.Laboratory of General Pathology and Experimental Therapy, Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 81, No. 3, pp. 259–261, March, 1976.     

Identification of a novel linear epitope on EspA from enterohemorrhagic E. coli using a neutralizing and protective monoclonal antibody

Enterohemorrhagic E. coli (EHEC) causes severe diseases in humans and animals via the production of Shiga toxins, and injection of effectors into epithelia using type III secretion system (TTSS). E. coli secreted protein A (EspA) forms the filamentous conduits of TTSS, which extends into the translocation pore embedded in host cell membranes and aids in the transportation of bacterial effectors. In addition, EspA is closely associated with initial bacterial adhesion and the formation of biofilms. EspA in its various forms elicits protective immune responses, although the epitope responsible has not to be identified. Here we report the presence of a linear, immunogenic, conserved and partially protective epitope E07 (100Lys-120Val) on EspA, which is recognized by the novel monoclonal antibody 1H10. This antibody blocks EHEC-induced actin polymerization and confers protection in mice. These findings provide a better understanding of EspA-induced immune responses and could lead to epitope-based vaccines and antibody-based therapies.     

Simulated microgravity affects ciprofloxacin susceptibility and expression of acrAB-tolC genes in E. coli ATCC25922

As a representative fluoroquinolone antibacterial, ciprofloxacin is frequently used to treat infections caused by bacteria such as E. coli. It is much meaningful to explore ciprofloxacin susceptibility and investigate a possible mechanism of drug susceptibility changes in E. coli ATCC25922 exposed to the environmental stress of simulated microgravity. The subculture of E. coli lasted for 7 days under simulated microgravity conditions (SMG) and normal microgravity (NG) conditions. On the 8th day, the cultures were divided into three groups: (1) NG group (continuous NG cultures); (2) SMG group (continuous SMG cultures); (3) SMCNG group (simulated microgravity change into normal gravity cultures). Ciprofloxacin (a final concentration of 0.125 μg/ml) sensitivity and expression of acrAB-tolC genes were detected in E. coli cells. The count and percentage of viable cells in the SMG cultures bacteria exposed to ciprofloxacin were higher than that in NG cultures and reduced to the levels of NG group when they were subcultivated from SMG to NG. The expressions of efflux pump genes (acrA, acrB and tolC) were upregulated in SMG culture and downregulated to the levels of NG group when they were subcultivated from SMG to NG. Susceptibility to ciprofloxacin and expression of acrAB-tolC genes in E. coli could be reversibly affected by SMG conditions. Over expression of efflux pump genes acrAB-tolC perhaps played an important role in decreased CIP susceptibility under SMG.     

Bacterial aetiology of diarrhoea in young children: high prevalence of enteropathogenic Escherichia coli (EPEC) not belonging to the classical EPEC serogroups

Diarrhoeal disease continues to be one of the most common causes of admittance in Children hospital emergency. The aim of the present study was to investigate the relative contribution of enteropathogenic and enterohemorrhagic Escherichia coli (EPEC and EHEC, respectively) as a cause of infectious bacterial diarrhoea in children from the region of Toulouse. We analysed 280 samples of stools from 280 children (<2 years) with diarrhoea admitted in the "Hopital des Enfants" from January to August 2005. Classic pathogens (Salmonella, Campylobacter, Yersinia, Shigella, Aeromonas and Vibrio) were detected by standard culture methods. Enterotoxigenic Clostridium difficile were identified after culture by immuno-enzyme assay (IEA). Virulence genes of EPEC and EHEC were detected by using PCR. Shiga-toxin production of EHEC strains was confirmed with an IEA test. Potential enteric pathogens were identified in 55 patients. EPEC was the most frequently identified agent (30 patients), followed by Campylobacter (9 cases: 7 C. jejuni and 2 C. coli) and C. difficile (8 patients), then EHEC (5 patients) and Salmonella (3 patients). No Shigella, Yersinia, Aeromonas or other pathogenic bacteria were detected during this period in that class of children. EPEC not belonging to the classical EPEC serogroups were highly prevalent (24 versus 6). EHEC possessed different genotypes and serogroups: O26 (2 strains), O157 (2 strains) and one un-typable strain. This study demonstrates the importance of EPEC (55 % of positive cases) and of EHEC (more frequent than Salmonella) in the aetiology of diarrhoeal diseases of young children. We confirm the usefulness of the PCR methodology: it allows the detection of virulent E. coli and thus increases by two fold the diagnosis of bacterial diarrhoea.     

Enteropathogenic Escherichia coli changes distribution of occludin and ZO-1 in tight junction membrane microdomains in vivo

Diarrhea is a disease caused by enteropathogenic Escherichia coli (EPEC) infection, which caused the deaths of several hundred thousand children each year. However, the molecular mechanisms underlying EPEC infection in vivo are not fully understood. In the present study, we used the C57BL/6J mouse as an in vivo model of EPEC infection and investigated the effect of EPEC on tight junction (TJ) structure and barrier function. TJ ultrastructure was studied by transmission electron microscopy and a small molecule tracer biotin was used to examine the paracellular permeability of the colon. The distribution of TJ proteins occludin and ZO-1 in the epithelium was investigated by immunofluorescence microscopy. Our results demonstrated that TJ structure was disrupted following EPEC infection. And the morphological changes of TJ were accompanied by increased paracellular permeability which led to impairment of TJ barrier function. Immunofluorescency analysis revealed that occludin and ZO-1 were translocated from villous membrane to the cytoplasm in intestinal epithelial cells during EPEC invasion. Moreover, wild-type EPEC and the mutant EPEC strain, ΔespF, had similar effects on barrier function and TJ protein localization at 5 days postinfection. Our findings demonstrate that EPEC infection in vivo led to disruption of tight junction barrier function. These results may provide insights into the molecular mechanism of the pathogenesis of EPEC infection.     

Enhanced vulnerability for Streptococcus pneumoniae sepsis during asplenia is determined by the bacterial capsule

Patients without a spleen are susceptible for overwhelming sepsis with Streptococcus pneumoniae. We investigated the relative contribution of the pneumococcal capsule in the reduced host defense after splenectomy.Sham-operated or splenectomized mice were inoculated with serotype 2 or 4 S. pneumoniae (D39, TIGR4) or the isogenic nonencapsulated mutants (D39Δcps, TIGR4Δcps). After splenectomy, intranasal infection with D39 resulted in increased mortality, increased bacterial dissemination and exaggerated systemic inflammation rather then altering inflammation in the lungs. Intravenous infection also resulted in enhanced mortality, bacterial growth and systemic inflammation after splenectomy. In contrast, the spleen did not contribute to host defense during infection with D39Δcps. Similar observations were made for TIGR4: increased bacterial growth and inflammation after intravenous infection with wild-type, but not nonencapsulated bacteria in splenectomized mice.These results indicate that the capsule of S. pneumoniae is indeed responsible for increased vulnerability of asplenic mice to invasive pneumococcal disease.     

Effects of natural honey on polymicrobial culture of various human pathogens

Introduction

Honey has a wide range of antimicrobial activity. All previous studies have considered honey''s effect on a single microbe. The present study investigated activity of honey towards a high dose of single or polymicrobial culture.

Material and methods

10 µl specimens of Staphylococcus aureus (S. aureus), Streptococcus pyogenes (S. pyogenes), Escherichia coli (E. coli) and Candida albicans (C. albicans) were cultured in 10 ml of 10-100% (wt/v) honey diluted in broth. Six types of polymicrobial microbial cultures were prepared by culturing the isolates with each other onto broth (control) and broth containing various concentrations of honey (10-100% wt/v). Microbial growth was assessed on solid plate media after 24 h incubation.

Results

Honey (30-70%) prevents growth of 10 µl specimens of all the isolates. Greater reduction in growth of E. coli was observed when cultured with S. aureus. Culturing of S. aureus with S. pyogenes, C. albicans, or E. coli increased its sensitivity to honey. S. aureus and S. pyogenes increased sensitivity of C. albicans to honey while E. coli and C. albicans decreased sensitivity of S. pyogenes.

Conclusions

It might be concluded that honey prevents and inhibits growth of single and polymicrobial pathogenic cultures. Polymicrobial culture affects growth of the isolates and increases their sensitivity to honey.     

Experimental Study on the Optimization of Extraction Process of Garlic Oil and its Antibacterial Effects

Background

Garlic oil which is the main active constituent of garlic has a wide range of pharmacological activities, and a broad antibacterial spectrum. It also has a strong anti-cancer activity, and can significantly inhibit a variety of tumors such as liver cancer, gastric cancer and colon cancer. The objective is to study the extraction process of garlic oil and its antibacterial effects.

Materials and Methods

CO₂ Supercritical extraction was used to investigate the optimal processing conditions for garlic oil extraction; filter paper test and suspension dilution test were applied to determine the bacteriostatic action of garlic oil.

Results

In the CO₂ supercritical extraction experiment, factors influencing the yield of garlic oil were: extraction pressure > extraction temperature > extraction time in descending order. Range analysis showed that the optimal experimental conditions for CO₂ supercritical extraction of garlic oil were extraction pressure of 15 Mpa, temperature of 40 °C, and duration of 1 h. Different concentrations of garlic oil could all inhibit the growth of Staphylococcus aureus, Escherichia coli and Bacillus subtilis, suggesting that garlic oil has an antibacterial effect.

Conclusion

The optimal experimental conditions for CO₂ supercritical extraction of garlic oil were: extraction pressure of 15 Mpa, temperature of 40 °C, and duration of 1 h; garlic oil has an antibacterial effect.     

Intersubunit signaling in RecBCD enzyme, a complex protein machine regulated by Chi hot spots

The Escherichia coli RecBCD helicase-nuclease, a paradigm of complex protein machines, initiates homologous genetic recombination and the repair of broken DNA. Starting at a duplex end, RecBCD unwinds DNA with its fast RecD helicase and slower RecB helicase on complementary strands. Upon encountering a Chi hot spot (5'-GCTGGTGG-3'), the enzyme produces a new 3' single-strand end and loads RecA protein onto it, but how Chi regulates RecBCD is unknown. We report a new class of mutant RecBCD enzymes that cut DNA at novel positions that depend on the DNA substrate length and that are strictly correlated with the RecB:RecD helicase rates. We conclude that in the mutant enzymes when RecD reaches the DNA end, it signals RecB's nuclease domain to cut the DNA. As predicted by this interpretation, the mutant enzymes cut closer to the entry point on DNA when unwinding is blocked by another RecBCD molecule traveling in the opposite direction. Furthermore, when RecD is slowed by a mutation altering its ATPase site such that RecB reaches the DNA end before RecD does, the length-dependent cuts are abolished. These observations lead us to hypothesize that, in wild-type RecBCD enzyme, Chi is recognized by RecC, which then signals RecD to stop, which in turn signals RecB to cut the DNA and load RecA. We discuss support for this "signal cascade" hypothesis and tests of it. Intersubunit signaling may regulate other complex protein machines.     

Experimental immunization with anti-rheumatic bacterial extract OM-89 induces T cell responses to heat shock protein (hsp)60 and hsp70; modulation of peripheral immunological tolerance as its possible mode of action in the treatment of rheumatoid arthritis (RA)

OM-89 is a bacterial (Escherichia coli) extract used for oral administration in the treatment of RA. Given the evidence that immunity to bacterial heat shock antigens plays a critical role in the immunomodulation of arthritis and possibly inflammation in general, the purpose of the present studies was to evaluate the presence and immunogenicity of hsp in OM-89. Furthermore, we studied the effects of OM-89 in an experimental arthritis, where hsp are known to have a critical significance in disease development. In rats immunization with OM-89 was found to lead to proliferative T cell responses to hsp60 and hsp70 of both E. coli and mycobacterial origin. Conversely, immunization with hsp antigens was also found to induce T cell reactivity specific for OM-89. Based on this and the antigen specificity analysis of specific T cell lines, hsp70 (DnaK) turned out to be one of the major immunogenic constituents of OM-89. Parenteral immunization with OM-89 was found to reduce resistance to adjuvant arthritis (AA), whereas oral administration was found to protect against AA. Given the arthritis-inhibitory effect of oral OM-89 in AA, it is possible that peripheral tolerance is induced at the level of regulatory T cells with specificity for hsp. This may also constitute a mode of action for OM-89 as an arthritis-suppressive oral drug.     

Evaluation of Phytochemical and Antibacterial Potential of Helicteres Isora L. Fruits Against Enteric Bacterial Pathogens

Antibacterial activities of aqueous, acetone, ethanol and methanol extracts of fruits of Helicteres isora (Mororphali) were studied. The fruit aqueous extracts of H. isora showed prominent antibacterial activities against E.coli, Staphylococcus epidermidis, Salmonella typhimurium and Proteus vulgaris; moderate activity against Enterobacter aerogenes, Staphylococcus aureus, Salmonella typhi and least activity against Pseudomonas aeruginosa. The aqueous extract showed maximal, the ethanol and methanol extract moderate and acetone extracts least antibacterial activities. Phytochemical screening revealed the presence of carbohydrates, anthraquinon glycosides, proteins, tannin and phenolic compounds and steroids These antibacterial properties supports its traditional use of fruits of H. isora in the treatment of enteric or diarrhoeal infections.     

Molecular evolution of the nicotinic acid requirement within the Shigella/EIEC pathotype

Nicotinamide adenine dinucleotide (NAD) is a crucial cofactor in several anabolic and catabolic reactions. NAD derives from quinolinic acid (QUIN) which in Escherichia coli is obtained through a pyridine salvage pathway or a de novo synthesis pathway. In the latter case, two enzymes, l-aspartate oxidase (NadB) and quinolinate synthase (NadA), are required for the synthesis of QUIN. In contrast to its E. coli ancestor, Shigella spp., the causative agent of bacillary dissentery, lacks the de novo pathway and strictly requires nicotinic acid for growth (Nic⁻ phenotype). This phenotype depends on the silencing of the nadB and nadA genes and its pathoadaptive nature is suggested by the observation that QUIN attenuates the Shigella invasive process. Shigella shares the pathogenicity mechanism with enteronvasive E. coli (EIEC), a group of pathogenic E. coli. On the basis of this similarity EIEC and Shigella have been grouped into a single E. coli pathotype. However EIEC strains do not constitute a homogeneous group and do not possess the complete set of characters that define Shigella strains.     

Changes in the microcirculation during the combined action of clostridium perfringens type a toxin and metabolic products of clostridium butyricum

In poisoning caused by injection ofClostridiumperfringens type A toxin or a mixture of toxin with the filtrate of a culture ofClostridium butyricum, changes in the microcirculation in the mucous membrane of the retrobuccal pouch of golden hamsters were found. The microcirculatory changes took place in two phases. In the first phase vasomotor disturbances were observed, as shown by periodic changes in diameter of the arterial microvessels, and by plasmatization and a decrease in the number of functioning capillaries. The diameter of the veins showed no significant change. The second phase was characterized by persistent disturbances of the microcirculation: slowing of the blood flow in the arterial and venous portions, the appearance of regions of stasis, a retrograde blood flow, and arterial dilatation. The persistent disturbances of the microcirculation following injection of a mixture ofCl. perfringens toxin and filtrate of a broth culture ofCl. butyricum appeared sooner than those in response to injection of the toxin alone.Department of Microbiology, N. I. Pirogov Odessa Medical Institute. Laboratory of Pathophysiology of Extremal States, Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. N. Chernukh.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 11, pp. 552–554, November, 1977.     

Virulence characteristics of translocating Escherichia coli and the interleukin-8 response to infection

Four efficiently translocating Escherichia coli (TEC) strains isolated from the blood of humans (HMLN-1), pigs (PC-1) and rats (KIC-1 and KIC-2) were tested for their ability to adhere and translocate across human gut epithelial Caco-2 and HT-29 cells, to elicit a proinflammatory response and for the presence of 47 pathogenic E. coli virulence genes. HMLN-1 and PC-1 were more efficient in adhesion and translocation than rat strains, had identical biochemical phenotype (BPT) and serotype (O77:H18) and phylogenetic group (D). KIC-2 adhered more than KIC-1, belonged to different BPT and serotype but the same phylogenetic group as KIC-1. TEC strains elicited significantly higher IL-8 response in both cell lines (P < 0.05) and monocytic THP-1 (P < 0.0001) cells than non-TEC strains. KIC-2 induced the highest IL-8 response which may be associated with its immunostimulatory flagellin. Apart from adhesin genes fimH and bmaE that were carried by all strains, HMLN-1 and PC-1 carried capsule synthesis gene kpsMT III and KIC-2 carried the EAST1 toxin gene. The lack of known virulence genes and the ability of TEC to efficiently adhere and translocate whilst causing proinflammatory response suggests that these strains may carry as yet unidentified genes that enable their translocating ability.