Variability in cell division among anatomical sites shapes Escherichia coli antibiotic survival in a urinary tract infection mouse model.

Urinary tract infection (UTI), mainly caused by Escherichia coli, are frequent and have a recurrent nature even after antibiotic treatment. Potential bacterial escape mechanisms include growth defects, but probing bacterial division in vivo and establishing its relation to the antibiotic response remain challenging. Using a synthetic reporter of cell division, we follow the temporal dynamics of cell division for different E. coli clinical strains in a UTI mouse model with and without antibiotics. We show that more bacteria are actively dividing in the kidneys and urine compared with the bladder. Bacteria that survive antibiotic treatment are consistently non-dividing in three sites of infection. Additionally, we demonstrate how both the strain in vitro persistence profile and the microenvironment impact infection and treatment dynamics. Understanding the relative contribution of the host environment, growth heterogeneity, non-dividing bacteria, and antibiotic persistence is crucial to improve therapies for recurrent infections.

Predictors of uncommon location of sentinel nodes in endometrial and cervical cancers.

OBJECTIVE: Sentinel node mapping is widely used in the treatment of gynecologic cancers. The current study aimed to identify predictors of uncommon sentinel lymph node (SLN) locations. METHODS: The current study included women who were operated for endometrial or cervical cancer with attempted sentinel lymph node mapping during surgical staging. Data were collected from electronic charts. The pelvis and the external ilia and obturator basins were common node locations. Para-aortic, pre-sacral, common iliac, internal iliac, and parametrial nodes were considered uncommon locations. We conducted analyses stratified according to common, uncommon, and very uncommon (para-aortic, pre-sacral, parametrial) node location sites. RESULTS: A total of 304 women were enrolled in the current study; 15.8% had SLN in uncommon locations and 4.3% had very uncommon node locations. Body mass index (BMI) was a negative predictor for uncommon SLN locations (OR 0.88, p = 0.03). The use of either indocyanine green (ICG) or Tc99 & blue dye was an independent predictor for uncommon SLN locations (OR 8.24, p = 0.006). More recent surgeries and the presence of positive nodes were independent predictors for very uncommon node locations (OR 2.13, p = 0.011, and OR 9.3, p = 0.002, respectively). CONCLUSIONS: BMI, tracer type, surgical year, and positive nodes were independent predictors for uncommon SLN locations. These findings suggest that surgical effort, technique and experience may result in better identification of uncommon SLN locations.

Major role of iron uptake systems in the intrinsic extra-intestinal virulence of the genus Escherichia revealed by a genome-wide association study.

The genus Escherichia is composed of several species and cryptic clades, including E. coli, which behaves as a vertebrate gut commensal, but also as an opportunistic pathogen involved in both diarrheic and extra-intestinal diseases. To characterize the genetic determinants of extra-intestinal virulence within the genus, we carried out an unbiased genome-wide association study (GWAS) on 370 commensal, pathogenic and environmental strains representative of the Escherichia genus phylogenetic diversity and including E. albertii (n = 7), E. fergusonii (n = 5), Escherichia clades (n = 32) and E. coli (n = 326), tested in a mouse model of sepsis. We found that the presence of the high-pathogenicity island (HPI), a ~35 kbp gene island encoding the yersiniabactin siderophore, is highly associated with death in mice, surpassing other associated genetic factors also related to iron uptake, such as the aerobactin and the sitABCD operons. We confirmed the association in vivo by deleting key genes of the HPI in E. coli strains in two phylogenetic backgrounds. We then searched for correlations between virulence, iron capture systems and in vitro growth in a subset of E. coli strains (N = 186) previously phenotyped across growth conditions, including antibiotics and other chemical and physical stressors. We found that virulence and iron capture systems are positively correlated with growth in the presence of numerous antibiotics, probably due to co-selection of virulence and resistance. We also found negative correlations between virulence, iron uptake systems and growth in the presence of specific antibiotics (i.e. cefsulodin and tobramycin), which hints at potential "collateral sensitivities" associated with intrinsic virulence. This study points to the major role of iron capture systems in the extra-intestinal virulence of the genus Escherichia.

Success of Escherichia coli O25b:H4 Sequence Type 131 Clade C Associated with a Decrease in Virulence.

Escherichia coli O25b:H4 sequence type 131 (ST131), which is resistant to fluoroquinolones and which is a producer of CTX-M-15, is globally one of the major extraintestinal pathogenic E. coli (ExPEC) lineages. Phylogenetic analyses showed that multidrug-resistant ST131 strains belong to clade C, which recently emerged from clade B by stepwise evolution. It has been hypothesized that features other than multidrug resistance could contribute to this dissemination since other major global ExPEC lineages (ST73 and ST95) are mostly antibiotic susceptible. To test this hypothesis, we compared early biofilm production, presence of ExPEC virulence factors (VFs), and in vivo virulence in a mouse sepsis model in 19 and 20 epidemiologically relevant strains of clades B and C, respectively. Clade B strains were significantly earlier biofilm producers (P < 0.001), carriers of more VFs (P = 4e-07), and faster killers of mice (P = 2e-10) than clade C strains. Gene inactivation experiments showed that the H30-fimB and ibeART genes were associated with in vivo virulence. Competition assays in sepsis, gut colonization, and urinary tract infection models between the most anciently diverged strain (B1 subclade), one C1 subclade strain, and a B4 subclade recombining strain harboring some clade C-specific genetic events showed that the B1 strain always outcompeted the C1 strain, whereas the B4 strain outcompeted the C1 strain, depending on the mouse niches. All these findings strongly suggest that clade C evolution includes a progressive loss of virulence involving multiple genes, possibly enhancing overall strain fitness by avoiding severe infections, even if it comes at the cost of a lower colonization ability.

Clinical Success Rate of Extensive Hysteroscopic Cesarean Scar Defect Excision and Correlation to Histologic Findings.

STUDY OBJECTIVE: Cesarean scar defect (CSD) is often associated with postmenstrual bleeding, infertility, and pain. Hysteroscopic CSD repair was described in the past, mainly as excision of the proximal edge of the defect to allow continuous blood flow during menstruation. In this study we aimed to evaluate the efficacy of extensive hysteroscopic cesarean scar niche excision in symptomatic patients. DESIGN: A retrospective cohort study. PATIENTS: Symptomatic patients treated with hysteroscopic CSD excision who were considered eligible for the procedure when myometrial thickness of 2 mm or more was observed on sonohysterography. SETTING: Tertiary referral center. INTERVENTIONS: Extensive CSD excision was performed using a cutting loop and pure cutting current. The proximal and distal edges of the defect were resected. This was followed by resection of tissue at the base of the niche, until underling muscular tissue was evident. Tissue sampled from the base of the CSD was collected for histologic examination. Patients were followed for a minimum of 1 year after hysteroscopic CSD excision. Clinical information obtained included detailed obstetric history and preoperative and postoperative menstruation pattern. MEASUREMENTS AND MAIN RESULTS: Between 2011 and 2016, 95 patients underwent extensive hysteroscopic niche excision; 67 were included in the study, whereas the remaining were lost to follow-up. Patient mean age at the time of the procedure was 38 ± 5.5 years. Twenty-nine patients (43%) had a history of high-order repeat cesarean surgeries. Sixty-six patients (98.5%) presented with postmenstrual bleeding, 26 with secondary infertility (38.8%), and 2 with pelvic pain (2.9%). After hysteroscopic niche excision, 63.4% of patients reported significant improvement or resolution of postmenstrual bleeding. A statistically significant reduction in number of bleeding days per cycle (15.5 ± 4.8 vs 9.8 ± 4.7, p < .001) was also noted. Histologic evidence for myometrial tissue within the obtained samples was associated with better outcomes. A histologic specimen from patients who experienced significant improvement or resolution of postmenstrual bleeding was more likely to reveal myometrial tissue (p = .04). Of the 26 patients who suffered from infertility, 19 attempted to conceive spontaneously after CSD excision. Of those, 10 patients (52.6%) conceived and 9 delivered at least once (47.36%). CONCLUSION: Extensive hysteroscopic surgical excision of cesarean scar niche should be considered in symptomatic patients suffering from irregular menstrual bleeding. The quality of the excision at the apex of the niche could be associated with a higher success rate. The role of niche excision to overcome secondary infertility should be further evaluated.

Control of pheromone production by ovaries in Drosophila.

Drosophila pheromones are long chain hydrocarbons (CHCs) produced by specialized epidermal cells, the oenocytes. Here we were explored the role of ovaries in CHC regulation. We studied tudor, a grandchildless-like mutation, resulting in progeny without ovaries and three alleles of ovoD, resulting in ovarian defects depending on the strength of the allele. We show here that these mutant flies with no or abnormal ovaries have a decrease in C29 length CHC ratio, balanced by an increase in C23 and C25 length ratio; this effect is dependent on the strength of the mutation. An increase in the amount of CHCs also occurred but was not related to the strength of ovoD alleles. As ovaries are the main site of ecdysone production in females, we knocked down the receptor to ecdysone EcR in the oenocytes and obtained increased amounts of CHCs and inhibition of long chain CHC synthesis, showing that the lack of an ecdysone signal arriving into the oenocytes is responsible for these defects. We then investigated the role of follicular cells and oocyte on CHC regulation: we RNAi-knocked down the LPR receptors in the oocyte to hinder vitellogenesis without significantly modifying CHC profile. We then expressed apoptosis genes within the follicle cells or within the ovocytes and obtained less long chain and more short chain CHC levels in the former case and an enhanced CHC production in the latter case. Together, these results support the notion of an interaction between oocyte and follicular cells, which send an ecdysone signal to the oenocytes to regulate CHC synthesis.

Feasibility and efficacy of repeated hysteroscopic cesarean niche resection.

OBJECTIVE: Cesarean-induced niche can cause symptoms such as abnormal postmenstrual bleeding, pain and associated infertility. Hysteroscopic niche resection is usually a successful treatment, but can result in a failure to improve symptoms or symptoms can recur. In the present study we aim to evaluate the feasibility, effectiveness, and safety of a second hysteroscopic niche resection for patients in whom an initial hysteroscopic resection failed to improve symptoms. STUDY DESIGN: This retrospective cohort study (Canadian Task Force classification II-2) hospital tell hashomer (tertiary center) included all patients who underwent a second hysteroscopic niche resection between 2011 and 2015. MEASUREMENTS: Fertility,obstetric outcomes, clinical outcome and complications were compared between the first surgery and the second RESULTS: Eight patients underwent a second hysteroscopy after failure of the first hysteroscopy to resolve symptoms or after recurrence of symptoms. Abnormal uterine bleeding (AUB) was the most common symptom, occurring in all patients. The average number of days of bleeding per cycle were significantly reduced following the second surgery [14.50 (range 8-21days) vs 11.75 (range 8-20days), respectivelyp=0.009]. The second surgery improved symptoms in 6 out of the 8 patients with AUB and 1 of 2 patients with pain. There were no significant differences in fertility and obstetric outcomes between the first and the second surgery and no complications were reported during any of the surgeries. CONCLUSION: Reintervention with a second hysteroscopic niche resection is both feasible and effective treatment option following a failed first attempt or recurrence of symptoms. The second surgery improved symptoms, especially AUB, with no consequent detrimental effect on obstetric outcomes on our series.

Acute cholecystitis: early versus delayed cholecystectomy, a multicenter randomized trial (ACDC study, NCT00447304).

OBJECTIVE: Acute cholecystitis is a common disease, and laparoscopic surgery is the standard of care. BACKGROUND: Optimal timing of surgery for acute cholecystitis remains controversial: either early surgery shortly after hospital admission or delayed elective surgery after a conservative treatment with antibiotics. METHODS: The ACDC ("Acute Cholecystitis-early laparoscopic surgery versus antibiotic therapy and Delayed elective Cholecystectomy") study is a randomized, prospective, open-label, parallel group trial. Patients were randomly assigned to receive immediate surgery within 24 hours of hospital admission (group ILC) or initial antibiotic treatment, followed by delayed laparoscopic cholecystectomy at days 7 to 45 (group DLC). For infection, all patients were treated with moxifloxacin for at least 48 hours. Primary endpoint was occurrence of predefined relevant morbidity within 75 days. Secondary endpoints were as follows: (1) 75-day morbidity using a scoring system; (2) conversion rate; (3) change of antibiotic therapy; (4) mortality; (5) costs; and (6) length of hospital stay. RESULTS: Morbidity rate was significantly lower in group ILC (304 patients) than in group DLC (314 patients): 11.8% versus 34.4%. Conversion rate to open surgery and mortality did not differ significantly between groups. Mean length of hospital stay (5.4 days vs 10.0 days; P < 0.001) and total hospital costs (€2919 vs €4262; P < 0.001) were significantly lower in group ILC. CONCLUSIONS: In this large, randomized trial, laparoscopic cholecystectomy within 24 hours of hospital admission was shown to be superior to the conservative approach concerning morbidity and costs. Therefore, we believe that immediate laparoscopic cholecystectomy should become therapy of choice for acute cholecystitis in operable patients. (NCT00447304).

Periodontopathic microorganisms in peripheric blood after scaling and root planing.

AIM: The objective of this study was to evaluate the frequency of periodontopathic and other subgingival anaerobic and facultative bacteria in the bloodstream following scaling and root planing (SRP). MATERIAL AND METHODS: Forty-two patients with severe generalized chronic periodontitis (GChP) and generalized aggressive periodontitis (GAgP) were included in the study. Four samples of peripheric blood were drawn from the cubital vein at different times: Pre-treatment: immediately before the SRP procedure (T1), immediately after treatment (T2), 15 min. post-treatment (T3) and 30 min. post-treatment (T4). In order to identify the presence of microorganisms in blood, subcultures were conducted under anaerobic conditions. RESULTS: 80.9% of the patients presented positive cultures after SRP and it occurred more frequently immediately after treatment; however, 19% of the patients still had microorganisms in the bloodstream 30 min. after the procedure. The periodontopathic microorganisms more frequently identified were Porphyromonas gingivalis and Micromonas micros. Campylobacter spp., Eikenella corrodens, Tannerella forsythensis, Fusobacterium spp. and Prevotella intermedia were isolated less often. Actinomyces spp. were also found frequently during bacteraemia after SRP. CONCLUSIONS: SRP induced bacteraemia associated with anaerobic bacteria, especially in patients with periodontal disease.

[Subgingival microbiota in chronic and aggressive periodontitis in Bogotá, Colombia: an epidemiological approach]. / Microflora subgingival en periodontitis crónica y agresiva en Bogotá, Colombia: un acercamiento epidemiológico.

INTRODUCTION: Periodontitis-associated microbiotas differ in regions around the world and vary with a variety of factors. This suggests that each country must establish its own dental microbiobiotic profile in order to formulate adequate prevention measures and more specific treatments. OBJECTIVE: The prevalence and proportions of the following microorganisms were investigated: Porphyromonas gingivalis, Tannerella forsythia, Actinobacillus actinomycetemcomitans, Prevotella intermedia/Prevotella nigrescens, Eikenella corrodens, Campylobacter rectus, Micromonas micros, Fusobacterium spp, Dialister pneumosintes and enteric rods. The patients selected had been diagnosed with chronic periodontitis and aggressive periodontitis from clinical populations in Bogotá, Colombia. The variables of age, gender and socioeconomic level were included in the comparisons. MATERIALS AND METHODS: 183 patients were classified in two groups, chronic periodontitis (n=84) and aggressive periodontitis (n=59); 40 healthy subjects were selected for comparison. A pooled subgingival plaque sample was obtained from the 6 deepest periodontal pockets in every patient and processed by culture for the isolation and identification of bacteria. RESULTS: In chronic periodontitis and aggressive periodontitis, P. gingivalis, T. forsythia, P. intermedia/P. nigrescens, C. rectus, Fusobacterium spp and E. corrodens showed high and similar prevalence, whereas M. micros, A. actinomycetemcomitans, D. pneumosintes and enteric rods (commonly from the tribe Klebsielleae) showed less and similar prevalence. In the healthy group, P. intermedia/P. nigrescens, C. rectus and Fusobacterium spp showed high frequency whereas M. micros and E. corrodens showed moderate frequency. CONCLUSIONS: Porphyromonas gingivalis, T. forsythia, P. intermedia/P. nigrescens, C. rectus, Fusobacterium spp. and E. corrodens are important constituents of the microbiologic profile of the periodontitis in Colombian populations. Porphyromonas gingivalis was found more frequently than A. actinomycetemcomitans in aggressive periodontitis.

Demographic, clinical, and microbial aspects of chronic and aggressive periodontitis in Colombia: a multicenter study.

BACKGROUND: The microbial profile of periodontal disease varies among different human populations. This study evaluated the demographic, clinical, and microbiologic aspects of periodontitis in a multigeographic sample in Colombia. METHODS: Three hundred twenty-five patients with chronic periodontitis (CP), 158 patients with aggressive periodontitis (AgP), and 137 healthy-gingivitis controls from five regions of the country were studied. Clinical, microbial, and sociodemographic data were collected. Microbiologic identification was performed using polymerase chain reaction 16S rRNA gene on pooled subgingival samples, and the presence of Gram-negative enteric rods was evaluated by culture. Bivariate and multivariate logistic regression analyses were conducted. RESULTS: Porphyromonas gingivalis occurred in 71.5% of individuals with periodontitis, Tannerella forsythensis occurred in 58.5%, Campylobacter rectus occurred in 57.5%, Actinobacillus actinomycetemcomitans occurred in 23.6%, and enteric rods occurred in 34.5%. P. gingivalis was more common in CP and AgP than controls. A. actinomycetemcomitans was increased in AgP compared to controls and patients with CP. T. forsythensis, C. rectus, and Eikenella corrodens had a low presence in the West Pacific and Central regions, and enteric rods were increased in the Central region (P <0.05). Other sociodemographic factors were not associated with these microorganisms. CONCLUSIONS: Geographic regions do not influence the microbiota, but the microbiota may vary by geographic region. P. gingivalis, T. forsythensis, and C. rectus are the most prevalent periodontophatic microorganisms in Colombia. A. actinomycetemcomitans was more common in AgP, and a large percentage of the population studied had enteric rods in the subgingival plaque.

Microflora subgingival en periodontitis crónica y agresiva en Bogotá, Colombia: un acercamiento epidemiológico / Subgingival microbiota in chronic and aggressive periodontitis in Bogotá, Colombia: an epidemiological approach

Introducción. Los microorganismos involucrados en la periodontitis varían en cada región dependiendo de diferentes factores. Por ello cada país debe establecer su propio perfil microbiológico con el objeto de proponer esquemas adecuados de prevención y tratamiento. Objetivo. Investigar la presencia y concentración subgingival de Porphyromonas gingivalis, Tannerella forsythia, Actinobacillus actinomycetemcomitans, Prevotella intermedia/Prevotella nigrescens, Eikenella corrodens, Campylobacter rectus, Micromonas micros, Fusobacterium spp , Dialister pneumosintes y bacilos entéricos en pacientes con periodontitis en una población colombiana considerando edad, género y nivel socioeconómico. Materiales y métodos. Participaron 183 pacientes: 84 con periodontitis crónica, 59 con periodontitis agresiva y 40 individuos sanos. De cada paciente se obtuvo un grupo de muestras de los seis sitios con mayor profundidad de bolsa que se procesaron por cultivo para el aislamiento e identificación de las bacterias. Resultados. En periodontitis crónica y agresiva se detectaron con alta y similar frecuencia P. gingivalis, T. forsythia, P. intermedia/P. nigrescens, C. rectus, Fusobacterium spp y E. corrodens y con menor y similar frecuencia, M. micros, A. actinomycetemcomitans, D. pneumosintes y bacilos entéricos, principalmente de la tribu Klebsielleae. En los sujetos sanos se encontraron con frecuencia elevada P. intermedia/P. nigrescens, C. rectus y Fusobacterium spp y con frecuencia moderada, M. micros y E. corrodens. Conclusión. Los resultados indican que P. gingivalis, T. forsythia, P. intermedia/P. nigrescens, C. rectus, Fusobacterium spp y E. corrodens constituyen parte importante del perfil microbiológico de las periodontitis en la población colombiana. En la periodontitis agresiva, P. gingivalis se encontró más frecuentemente que A. actinomycetemcomitans.

Introduction. Periodontitis-associated microbiotas differ in regions around the world and vary with a variety of factors. This suggests that each country must establish its own dental microbiobiotic profile in order to formulate adequate prevention measures and more specific treatments. Objective. The prevalence and proportions of the following microorganisms were investigated: Porphyromonas gingivalis, Tannerella forsythia, Actinobacillus actinomycetemcomitans, Prevotella intermedia/Prevotella nigrescens, Eikenella corrodens, Campylobacter rectus, Micromonas micros, Fusobacterium spp, Dialister pneumosintes and enteric rods. The patients selected had been diagnosed with chronic periodontitis and aggressive periodontitis from clinical populations in Bogotá, Colombia. The variables of age, gender and socioeconomic level were included in the comparisons. Materials and methods. 183 patients were classified in two groups, chronic periodontitis (n=84) and aggressive periodontitis (n=59); 40 healthy subjects were selected for comparison. A pooled subgingival plaque sample was obtained from the 6 deepest periodontal pockets in every patient and processed by culture for the isolation and identification of bacteria. Results. In chronic periodontitis and aggressive periodontitis, P. gingivalis, T. forsythia, P. intermedia/P. nigrescens, C. rectus, Fusobacterium spp and E. corrodens showed high and similar prevalence, whereas M. micros, A. actinomycetemcomitans, D. pneumosintes and enteric rods (commonly from the tribe Klebsielleae) showed less and similar prevalence. In the healthy group, P. intermedia/P. nigrescens, C. rectus and Fusobacterium spp showed high frequency whereas M. micros and E. corrodens showed moderate frequency. Conclusions. Porphyromonas gingivalis, T. forsythia, P. intermedia/P. nigrescens, C. rectus, Fusobacterium spp. and E. corrodens are important constituents of the microbiologic profile of the periodontitis in Colombian populations...