Plasmids carrying antimicrobial resistance genes in Enterobacteriaceae.

Bacterial antimicrobial resistance (AMR) is constantly evolving and horizontal gene transfer through plasmids plays a major role. The identification of plasmid characteristics and their association with different bacterial hosts provides crucial knowledge that is essential to understand the contribution of plasmids to the transmission of AMR determinants. Molecular identification of plasmid and strain genotypes elicits a distinction between spread of AMR genes by plasmids and dissemination of these genes by spread of bacterial clones. For this reason several methods are used to type the plasmids, e.g. PCR-based replicon typing (PBRT) or relaxase typing. Currently, there are 28 known plasmid types in Enterobacteriaceae distinguished by PBRT. Frequently reported plasmids [IncF, IncI, IncA/C, IncL (previously designated IncL/M), IncN and IncH] are the ones that bear the greatest variety of resistance genes. The purpose of this review is to provide an overview of all known AMR-related plasmid families in Enterobacteriaceae, the resistance genes they carry and their geographical distribution.

Spread of mcr-1-carrying Enterobacteriaceae in sewage water from Spain.

Objectives: The mobile colistin resistance gene mcr-1 has been identified worldwide in human and animal sources, while its occurrence in the environment is still largely unknown. The aim of this study was to investigate the presence of mcr-1 -harbouring Enterobacteriaceae in water samples obtained from rivers and waste water treatment plants in the area of Barcelona, Spain. Methods: The presence of mcr-1 was detected by PCR. Bacterial identification was performed via MALDI-TOF MS. Resistance to colistin was determined by a broth dilution method. The epidemiological relationship between the positive isolates was assessed with PFGE and ST was determined by MLST. Plasmid characterization was performed by transformation experiments, antimicrobial susceptibility testing and incompatibility group PCR. Results: Thirty MDR isolates bearing mcr-1 , 29 Escherichia coli (ST632 and ST479) and 1 Klebsiella pneumoniae (ST526), were identified in sewage from two different waste water treatment plants, whereas the gene was not found in river water. All isolates, including the K. pneumoniae , harboured bla CTX-M-55 and bla TEM-1 . mcr-1 was in all cases associated with an IncI2 plasmid, which only conferred resistance to colistin. mcr-1 was harboured by two predominant E. coli clones that were found in both waste water treatment plants. Conclusions: This study showed a high occurrence of mcr-1 in the sewage of Barcelona, mainly due to the dissemination of two E. coli pulsotypes that are circulating in the population. The presence of mcr-1 in the environment is a cause for concern, and suggests high prevalence of mcr-1 in the community.

Reversible testosterone-induced azoospermia in a 45-year-old man attending an infertility outpatient clinic.

A 45-year-old male with azoospermia was presented to us with primary infertility. We found that he had been taking testosterone due to erectile dysfunction. Upon its discontinuation, the normozoospermia was restored. This led to a pregnancy. In men aged 40-50, the diagnostic work up of testosterone deficiency should be in accordance with the current guidelines. Moreover, no testosterone prescription should be made without having inquired about parenthood desires. In men of the reproductive age with recent-onset azoospermia, consider the possibility of an iatrogenic cause, primarily due to the testosterone replacement therapy. The treatment is simple: to discontinue the therapy.

Antimicrobial resistance and One Health.

Antimicrobial resistance is one of the major health problems we face in the 21st century. Nowadays we cannot understand global health without the interdependence between the human, animal and environmental dimensions. It is therefore logical to adopt a "One Health" approach to address this problem. In this review we show why a collaboration of all sectors and all professions is necessary in order to achieve optimal health for people, animals, plants and our environment.

Harmonisation of in-silico next-generation sequencing based methods for diagnostics and surveillance.

Improvements in cost and speed of next generation sequencing (NGS) have provided a new pathway for delivering disease diagnosis, molecular typing, and detection of antimicrobial resistance (AMR). Numerous published methods and protocols exist, but a lack of harmonisation has hampered meaningful comparisons between results produced by different methods/protocols vital for global genomic diagnostics and surveillance. As an exemplar, this study evaluated the sensitivity and specificity of five well-established in-silico AMR detection software where the genotype results produced from running a panel of 436 Escherichia coli were compared to their AMR phenotypes, with the latter used as gold-standard. The pipelines exploited previously known genotype-phenotype associations. No significant differences in software performance were observed. As a consequence, efforts to harmonise AMR predictions from sequence data should focus on: (1) establishing universal minimum to assess performance thresholds (e.g. a control isolate panel, minimum sensitivity/specificity thresholds); (2) standardising AMR gene identifiers in reference databases and gene nomenclature; (3) producing consistent genotype/phenotype correlations. The study also revealed limitations of in-silico technology on detecting resistance to certain antimicrobials due to lack of specific fine-tuning options in bioinformatics tool or a lack of representation of resistance mechanisms in reference databases. Lastly, we noted user friendliness of tools was also an important consideration. Therefore, our recommendations are timely for widespread standardisation of bioinformatics for genomic diagnostics and surveillance globally.

Changes in sperm apoptotic markers as related to seminal leukocytes and elastase.

To elucidate the effects of inflammation on sperm quality, this study analysed classical sperm characteristics, leukocytes and elastase in neat semen, and sperm apoptotic markers, i.e. changes in plasma membrane phospholipid asymmetry, mitochondrial membrane potential (MMP), DNA integrity and intracellular reactive oxygen species (ROS), in semen prepared by density gradient using flow cytometry from 348 men of infertile couples. Increased leukocytes (> or = 0.1 x 10(6)/ml) were associated with a decreased sperm concentration, motility and normal morphology (P < or = 0.001). Sperm necrosis and DNA denaturation were increased (31.3 versus 26.6%, P=0.020; 15.5 versus 11.5%, P = 0.011, respectively), whereas spermatozoa with normal MMP were decreased (64.1 versus 70.0%, P=0.004). High leukocyte levels ((> or = 1 x 10(6)/ml) were not associated with any of the observed sperm parameters. At low elastase concentration (100-290 microg/l), DNA denaturation was higher (16.1 versus 10.5%, P = 0.024) compared with very low elastase concentration (< 100 microg/l). A high elastase concentration (290-1000 microg/l) was associated with higher ROS index compared with low elastase concentration (1.28 versus 1.01, P=0.016). Slightly increased leukocytes and elastase are associated with slightly poorer sperm characteristics and/or increased sperm necrosis, DNA denaturation and intracellular ROS and decreased MMP.

Psychological factors in male partners of infertile couples: relationship with semen quality and early miscarriage.

In this study we sought to evaluate whether psychological factors in males affect semen quality and pregnancy. In 1076 men of infertile couples, psychological factors, i.e. exposure to acute stress, coping with stress, the WHO (five) Well-Being Index and the Zung's Anxiety Scale Inventory scores were assessed by a questionnaire at the time of semen analysis. Relationships between psychological factors and semen quality (sperm concentration, rapid and progressive motility and normal morphology) were assessed. In 353 men with infertility duration of < or =1.5 years, sperm concentration > or =5 x 10(6) sperm/mL and a female partner with a laparoscopically confirmed tubal patency, we looked prospectively for relations between psychological factors and the occurrence of a natural pregnancy at a 6-month follow-up (n = 124), and first-trimester loss (n = 18). Anxiety trait, found in 19% of men, was related to previous in vitro fertilization/intracytoplasmic sperm injection attempts (p = 0.014), cigarette intake (p = 0.006), alcohol intake (p = 0.026) and sexual difficulties (p < 0.001). Regression analyses indicated a significant positive relationship between the level of sperm concentration and the WHO (five) Well-Being Index score, each successive score number accounting for a 7.3% increase in sperm concentration (p = 0.039), whereas no correlation was found between psychological factors and sperm rapid progressive motility and normal morphology. Poorer coping with stress was related to the occurrence of a first-trimester miscarriage (p = 0.016) in the female partner. Possible depression in males is related to decreased sperm concentration, and poor coping with stress is associated with increased occurrence of early miscarriage.

Genetic variation in leptin and leptin receptor genes as a risk factor for idiopathic male infertility.

The aim of this study was to examine whether there is an association among genetic variability in leptin (LEP) and leptin receptor (LEPR) genes and male infertility. We performed a case-control study and were searching for an association between polymorphisms of LEP and LEPR genes and male infertility. The study group consisted of 317 patients with idiopathic infertility and a control group of 241 fertile men from Slovenia. Four single nucleotide polymorphisms (SNPs) in LEP gene and four single nucleotide polymorphisms (SNPs) in LEPR gene were chosen and genotyped. Statistically significant SNP was further validated in additional 255 infertile patients and 168 controls from Serbia and Macedonia. In the Slovenian population, we found a statistically significant difference in genotype distribution for rs10244329 polymorphism in LEP gene (recessive genotype model, p value = 0.048). The trend toward statistically significant difference in genotype distribution for rs10244329 polymorphism was confirmed in the Serbian and Macedonian populations (p value = 0.07). Our data suggest that genetic variability in the LEP gene might be associated with male infertility warranting further confirmation and mechanistic investigations.

[Sperm biochemical markers of genital tract inflammation: usefulness of elastase determination]. / Marqueurs biochimiques du sperme à la recherche de l'inflammation génitale: intérêt de la détermination de l'élastase.

Genital tract inflammation was for a long time suspected of inducing sterility in the male. The role of leukocytes in sperm changes is still debated. The identification of inflammation is based on the presence of activated leukocytes and the elastase measurement. Residual inflammation is evaluated by the reactive oxygen species activity. The true damage caused by inflammation on spermatozoa is reflected in negative changes observed during sperm apoptosis. Increased sperm DNA fragmentation is frequently observed while classical sperm characteristics are not modified automatically. The diagnosis of genital tract inflammation, follow-up after treatment and prognosis in terms of fertility require the use of different and complementary parameters.

Pathogenicity islands and virulence evolution in Listeria.

As in other bacterial pathogens, the virulence determinants of Listeria species are clustered in genomic islands scattered along the chromosome. This review summarizes current knowledge about the structure, distribution and role in pathogenesis of Listeria virulence loci. Hypotheses about the mode of acquisition and evolution of these loci in this group of Gram-positive bacteria are presented and discussed.

Cost-benefit analysis of pharmacologic erection program (PEP) versus penile prosthesis.

We compared the current dollar inclusive costs of the two most commonly implanted penile prostheses with our pharmacologic erection program (PEP). The inclusive cost of implantation for the inflatable penile prosthesis (IPP) is $9,000 and for the malleable penile prosthesis (MPP) $3,150. The inclusive cost of PEP at maximal dose and maximal frequency of use over a five-year period is $3,450. Penile prosthesis was found to be more cost-effective for the long-term treatment of younger impotent men, whereas in the elderly group the use of PEP over a limited time span may be equally or more cost-effective.

Factors associated with clinical assessment of overactive bladder and selection of treatment.

BACKGROUND: Overactive bladder (OAB) affects >17 million individuals in the United States, but the symptoms of OAB are frequently underreported by patients and therefore untreated by physicians. OBJECTIVE: The purpose of this observational study was to investigate the demographic and clinical factors associated with the decision to treat OAB pharmacologically and identify factors associated with physicians' assessment of symptom severity. METHODS: We studied 31 physicians treating 217 patients with OAB and collected data on patient demographic characteristics, OAB symptoms, previous management strategies, physicians' assessments of OAB severity, and treatment prescribed. Stepwise logistic regression was used to identify factors associated with selecting pharmacologic treatment and with physician assessment of severity of OAB symptoms. RESULTS: The mean age of the patients was 61.3 years; approximately 82% were female, and approximately 73% were white. Participants with urinary incontinence were significantly (P < 0.001) more likely to be treated with medication than were those with only symptoms of nocturia or urinary frequency. Other significant factors associated with pharmacologic treatment were being white (odds ratio [OR], 9.5; 95% CI, 2.9-30.8); being black (OR, 5.9; 95% CI, 1.2-29.7); physician's clinical assessment of OAB as moderate (OR, 3.5; 95% CI, 1.5-8.2) or severe (OR, 3.8; 95% CI, 1.1-13.7); previous use of medication (OR, 2.9; 95% CI, 1.1-8.1); and number of incontinence episodes in the last 24 hours (OR, 1.2; 95% CI, 1.0-1.5). Factors associated with physician assessment of OAB severity included distress due to OAB symptoms (OR, 2.1; 95% CI, 1.3-3.2), number of incontinence episodes in the last 24 hours (OR, 1.2; 95% CI, 1.1-1.4), and use of previous treatment(s) (OR, 0.4; 95% CI, 0.2-0.8). CONCLUSIONS: Both demographic and clinical symptoms of OAB were associated with the decision to treat OAB with medication, whereas physicians' assessment of OAB severity was associated only with clinical symptoms. Urinary incontinence was the key symptom associated with the decision to treat patients with medication and with the assessment of OAB symptom severity.

3-Desoxyanthocyanidins from Arrabidaea chica.

The new 3-desoxyanthocyanidins 6,7,3'-trihydroxy-5,4'-dimethoxy-flavylium and 6,7,3',4'-tetrahydroxy-5-methoxy-flavylium and the known 6,7-dihydroxy-5,4'-dimethoxy-flavylium (Carajurin) were isolated by bioguided fractionation from the leaves of Arrabidaea chica, with transcription factor NF-kappaB as target. The structure of Carajurone was revised to be 6,7,4'-trihydroxy-5-methoxy-flavylium. Additionally, the flavone acacetin was found. All structures were mainly established on the basis of MS- and NMR data (1H, 1H-1H COSY and partly 13C, GHMQCR and GHSQCR). Carajurin, which failed to give a positive result in the DPPH TLC assay completely inhibited NF-kappaB, but not NF-AT at a 500 microM concentration.

Pentosan polysulfate decreases prostate smooth muscle proliferation and extracellular matrix turnover.

Benign prostatic hyperplasia (BPH) involves proliferation of smooth muscle cells and increased deposition of extracellular matrix (ECM). We recently found that pentosan polysulfate (PPS) has marked effects on growth and ECM of smooth muscle cells derived from vascular tissues. We examined smooth muscle cells cultured from human prostates and the effects of PPS on their growth and ECM production. Fragments of surgical prostatectomy specimens were diced, digested with collagenase (0.01%), and placed in culture medium supplemented with 20% fetal bovine serum. Outgrowths of elongated cells were characterized by light microscopic examination and immunohistochemical techniques by the presence of F-actin, alpha-smooth muscle actin, and myosin, which is a characteristic of smooth muscle cells. Two independent isolates were propagated, and growth curves and ECM production were assessed in the presence and absence of PPS (10 or 100 microg/ml). PPS decreased cell number beginning at day 1 and throughout the incubation period, up to 4 days. The amount of the ECM degradative enzymes, metallo-proteinases MMP-9 and MMP-2, was examined by zymography. PPS did not alter the amount of MMP-2 in the supernatants but MMP-9 was increased 234.4 +/- 17.23-fold over control cells. Tissue inhibitor of MMP (TIMPS), examined by reverse zymography, increased 200% over control. The amount of alpha I type (IV) and alpha I type (I) collagen released in the supernatant, measured by ELISA, significantly decreased in PPS-treated cultures. In conclusion, we found that the administration of PPS decreased proliferation as well as ECM production in prostate smooth muscle. Since smooth muscle proliferation and ECM are involved in the pathophysiology of BPH, PPS may have therapeutic potential.

Involvement of CFTR gene alterations in obstructive and nonobstructive infertility in men.

There have not been many studies concerning CFTR gene alterations in nonobstructive causes of male infertility and subfertility, and in those that have been published, the results reported are not concordant. Therefore, we proposed to determine, in a representative unselected sample of men who were sent for microsurgical epididymal sperm aspiration, if different types of male infertility and impaired fertility were associated with CFTR gene alterations. We screened 80 men with idiopathic azoospermia, 50 men with severe oligozoospermia, 70 men with oligoasthenoteratozoospermia, and 7 men with congenital bilateral absence of the vas deferens (CBAVD), as well as 95 controls from Slovenia, for mutations in 10 CFTR exons that include the majority of the most common cystic fibrosis (CF) disease causing mutations. We also wanted to evaluate the risk for CF in children born after the intracytoplasmic sperm injection (ICSI) method of in vitro fertilization (IVF). No tested individual had mutations in both CFTR alleles. Altogether 13 different nucleotide alterations were identified. The frequencies of both CFTR gene alterations and polymorphisms did not differ significantly between the control group and men with idiopathic nonobstructive azoospermia and subfertility, but were significantly increased in men with CBAVD (DeltaF508, p = 0.039; IVS8-5T, p = 0.006). Our results suggest that CFTR mutations are not associated with errors in spermatogenesis and nonobstructive pathology of urogenital tract in men with any frequency. However, genetic counseling and CFTR mutation screening continue to be recommended for men with obstructive azoospermic conditions and their female partners.

Benign prostatic hyperplasia.

Benign prostatic hyperplasia (BPH) is a pathologic disorder that develops in response to the action of dihydrotestosterone on the aging prostate and to changes in stromal and epithelial cells in this exocrine gland. The current therapies for this disorder are chosen after other causes for irritative and obstructive symptoms have been excluded and the status of the urinary tract has been assessed. This evaluation includes a detailed medical history, a thorough genitourinary and neurological examination, assessment of serum prostate specific antigen and creatinine levels, as well as a urinalysis. A urodynamic evaluation consisting of a combined pressure-flow study is required if the diagnosis of obstruction is to be made. Patients with minimal symptoms and normal test results require no therapy. Mild to moderate symptoms can be controlled, at least temporarily, with alpha-adrenergic blockers such as terazosin or doxazosin. A subset of BPH patients with obstructive symptoms respond to the 5 alpha-reductase inhibitor finasteride. Early results with minimally invasive treatments such as laser prostatectomies, hyperthermia, and ultrasonic and radiofrequency ablation appear encouraging for those with moderate symptoms of prostatism. Severe symptoms, urinary retention, gross hematuria, recurrent urinary tract infections, bladder calculi, and hydronephrosis or renal insufficiency warrant transurethral incision, resection, vaporization, or open prostatectomy (for very large neoplasms). Although the morbidities of these latter surgical therapies are not insignificant, these treatments offer the best and most durable results for relief of obstruction and amelioration of symptoms.

[Personality traits of men with Klinefelter syndrome and their partners]. / Traits de personnalité des hommes atteints de syndrome de Klinefelter et de leurs partenaires.

The aim of the study was to find whether personality traits of men with Klinefelter syndrome and their partners (group 1-n = 17) differ from those of couples affected by idiopathic infertility (group 2; n = 16) and from those of fertile couples (group 3 n = 17). We further investigated the attitudes of the three groups towards pregnancy, labour and sexuality to find potential différences among the three groups. Besides, we verified the hypotheses of below average or low average intelligence of men with Klinefelter syndrome, and of inferior quality of social life in these men. The data were collected using the interview on medical history, the questionnaire on attitudes towards pregnancy, labour and sexuality (S-S-G), the personality questionnaire MMPI-2. The Raven progressive matrices were used only in group 1. The results show that men with Klinefelter syndrome and their partners do not differ significantly from the couples with idiopathic infertility (group 2), having some shizoide traits in their personality structure and mostly negative attitudes towards pregnancy, labour and sexuality. However, a significant difference has been found between the Klinefelter syndrome group and the fertile couples group. The hypothesis of below average intelligence has not been confirmed, but the quality of social life of men with Klinefelter syndrome has been found inférior. We may thus conclude that in the management of infertile couples in whom the man has been affected by Klinefelter syndrome, the personality structure, importantly affecting the outcome of treatment, should be taken into consideration.

[Isolation of Chlamydia trachomatis from the endometrium and endometritis. Apropos of a series of 42 patients]. / Isolement de Chlamydia trachomatis dans l'endomètre et endométrite. A propos d'une série de 42 patientes.

Most studies have been concerned with recovering Chlamydia trachomatis (CT) from the endocervix on the one hand and from the adnexae on the other hand and consider its relationship to infection of the upper genital tract. Our work is concerned with the endometrium. 42 women were examined: 22 of them had salpingitis (group I), 14 were considered possibly to have endometritis with a risk of infection with CT, or had cervicitis (group II), and 6 were in the control group (group III). Samples were taken from the endometrium that had been removed for histological examination and for searching for CT in cultures. These samples were taken routinely at the initial examination and then again 10 and/or 40 days later for women in groups I and II. These latter were all treated systemically with cyclines. A high incidence of endometrial infection with CT was found (17%). The recovery of CT from the endometrium was correlated with the presence of an intra-uterine device, with purulent discharge, with evidence of CT in the cervix, and with anti-chlamydia serology higher than or equal to 1/128. CT infection of the endometrium gave late lesions of endometritis which persisted in spite of the antibiotic therapy that had been given.

[Congenital auriculo-ventricular block and anti-Ro (SS-A)/anti-La (SS-B) antibody]. / Bloc auriculo-ventriculaire congénital et anticorps anti-Ro (SS-A)/anti-La (SS-B).

Congenital heart blocks due to immunological causes are rare. A case is reported of a fetus with auriculo-ventricular block diagnosed at 22 weeks of amenorrhoea and intrauterine death at 32 weeks. The authors discussing the case find the most likely link: an anti-RO (SS-A) and anti-LA (SS-B) immunological block and they suggest that there are minor localised lesions in the nodal tissue which gives rise to benign disturbances of cardiac rhythm and they point out ways of preventing intrauterine auriculo-ventricular block.

[Adrenal fetus in fetus. A case shown by prenatal echography. Review of the literature]. / Foetus in foetu surrénalien: un cas évoqué par l'échographie prénatale. Revue de la littérature.

The authors report the first case of fetus in fetu situated in the adrenal gland where the diagnosis was suggested by prenatal ultrasound. The anatomo-pathology, the aetiology, the antenatal ultrasound picture and the treatment of this abnormality of embryonic development are described.