Development of a novel selective agar for the isolation and detection of Bacillus anthracis.

AIMS: The aim of this study was to develop a novel selective agar for the specific isolation and detection of Bacillus anthracis. METHODS AND RESULTS: Based on published data on antibiotic resistance and susceptibility of B. anthracis and other closely related species of the Bacillus cereus sensu lato group, a new selective agar formulation termed CEFOMA (Bacillus CEreus sensu lato group-specific antibiotics, FOsfomycin, MAcrolides) was developed and evaluated. All tested strains of B. anthracis were able to grow on CEFOMA with the same colony number as on non-selective media, whereas CEFOMA inhibited the growth of the other species within the B. cereus sensu lato group. In comparison to other selective agars, CEFOMA had a superior performance and considerably reduced the total amount of accompanying flora in soil. Furthermore, B. anthracis was successfully isolated from deliberately spiked soil samples. CONCLUSIONS: CEFOMA is a highly promising selective agar for the efficient isolation of B. anthracis from environmental samples with a large bacterial background flora. SIGNIFICANCE AND IMPACT OF THE STUDY: The isolation of B. anthracis from environmental samples is severely impaired by the lack of adequate selective agars which suppress the growth of other bacteria. CEFOMA agar represents an important improvement and suitable alternative to currently used selective agars.

CMR-guidance of passively tracked endomyocardial biopsy in an in vivo porcine model.

Endomyocardial biopsy (EMB) is considered to be the diagnostic gold-standard in detection of myocardial-inflammation. EMB is usually conducted under fluoroscopy without any specific target information. Specific target-information provided by cardiovascular magnetic resonance (CMR) may improve specificity of EMB. The aim was to investigate feasibility and safety of CMR-guided and targeted EMB in a preclinical-model using passively-tracked devices. Procedures were performed on a MRI-System equipped with an Interventional Software-Platform for real-time imaging. Ex vivo experiments were conducted to optimize visibility of the guide-sheath. In vivo experiments were conducted in 2 pigs for technical feasibility assessment and in 4 pigs after acute myocardial infarction to test feasibility of guided and lesion targeted EMB. For anatomical real-time imaging a single-shot-balanced-SSFP-sequence was applied. Myocardial targets were identified under real-time imaging (single-shot-T2 (sshT2) and single-shot Late-Gadolinium-Enhancement (sshLGE) sequences). Ex vivo experiments demonstrated best visibility of continuously labelled guide-sheath. CMR-guided EMB was feasible in all cases without major complications. Likewise, lesion-targeting endomyocardial biopsy was feasible in two cases. Biopsies exhibited appropriate sizes and qualities. Real-time lesion sequences revealed comparable CNR values to clinical-protocols. Real-time imaging of lesions showed following signal- and contrast-to-noise ratios (SNR/CNR): SNR of sshT2- and sshLGE was 124 ± 35 and 67 ± 51 respectively, whereas CNR was 81 ± 30 and 57 ± 44. This study demonstrates feasibility and safety of CMR-guided and basically targeted EMB with passively-tracked devices. Signal-to-noise ratios of real-time sequences is non-inferior to standard sequences for lesion detection. CMR-guidance may improve diagnostic accuracy of EMB since CMR can detect myocardial-targets under real-time-imaging.

In vitro susceptibility to 19 agents other than ß-lactams among third-generation cephalosporin-resistant Enterobacteriaceae recovered on hospital admission.

Objectives: As part of the multicentre Antibiotic Therapy Optimisation Study, MIC values of 19 non-ß-lactam agents were determined for third-generation cephalosporin-resistant Escherichia coli , Klebsiella species and Enterobacter species (3GCREB) isolates collected in German hospitals. Methods: A total of 328 E. coli , 35 Klebsiella spp. (1 Klebsiella oxytoca and 34 Klebsiella pneumoniae ) and 16 Enterobacter spp. (1 Enterobacter aerogenes and 15 Enterobacter cloacae ) isolates were submitted to broth microdilution antimicrobial susceptibility testing with the MICRONAUT system. MICs of fluoroquinolones (levofloxacin and moxifloxacin), aminoglycosides (gentamicin, tobramycin, amikacin, streptomycin, neomycin and paromomycin), tetracyclines (tetracycline, minocycline and tigecycline), macrolides (erythromycin, clarithromycin and azithromycin) and miscellaneous agents [trimethoprim/sulfamethoxazole, chloramphenicol, nitrofurantoin, colistin and fosfomycin intravenous (iv)] were determined and reviewed against 2016 EUCAST breakpoints. Results: The MIC of levofloxacin was >2 mg/L for 128 of 328 E. coli and 8 of 35 Klebsiella spp., but only 1 of 16 Enterobacter spp. Rates of resistance to trimethoprim/sulfamethoxazole were high (>70%), except for Enterobacter spp. Rates of resistance to colistin and fosfomycin iv were still low. About 20% of the tested isolates were resistant to chloramphenicol. Only 1 (of 328) E. coli isolate had an MIC of amikacin >16 mg/L and only 33 of 328 E. coli and 1 of 35 Klebsiella spp. had an MIC of tobramycin >4 mg/L, whereas average gentamicin MICs were in general more elevated. A tigecycline MIC >2 mg/L was only found for 1 of 16 Enterobacter spp., but in none of the E. coli or Klebsiella spp. isolates. Conclusions: Our study gives insight into previously unreported non-ß-lactam MIC distributions of 3GCREB isolates.

[Peripartum management plan for patients with mental illnesses : Strategies to reduce the risk of postpartum relapse]. / Peripartales Management bei psychischer Vorerkrankung : Strategien zur Rezidivprophylaxe nach der Entbindung.

BACKGROUND: Transition to parenthood is challenging but for women with a history of recurrent psychiatric disorders becoming a mother has a number of additional issues. Women with a history of mood disorders or psychoses are at increased risk for exacerbation in the vulnerable postpartum period and fear the potential risk of medication during pregnancy for the unborn child as much as they fear a relapse. In these difficult situations women and their families seek advice and support from mental health providers and obstetricians. METHODS AND RESULTS: Based on the treatment of 420 mentally ill women with a desire to have children and pregnancy and prospective documentation of 196 pregnancies over the last 10 years (2006-2016) the authors developed the Bonn concept of peripartum management (BKPM). The plan was designed to reduce the incidence and severity of postpartum relapses in women suffering from psychiatric disorders. Factors to be considered include antenatal and postpartum medication as well as reduction of stress and stimuli, sleep preservation, social support and help from the partner in caring for the baby. Of the 196 women in the BKPM only 4.6 % (n = 9) experienced a severe postpartum relapse with hospitalization. Additionally, the informed consent discussion with patient and partner as part of the peripartum management plan showed positive effects on how women and their families experienced autonomy and safety during pregnancy and postpartum. DISCUSSION: Careful planning and monitoring with a structured perinatal management plan can reduce the risk of relapse in the perinatal period and thus support women with a history of mental disorder in the transition to motherhood. Therefore, the management concept employed in Bonn contributes to the major goal of current peripartum psychiatric care in developing effective prevention strategies for women at high risk.

Colonization with third-generation cephalosporin-resistant Enterobacteriaceae on hospital admission: prevalence and risk factors.

OBJECTIVES: The objectives of this study were to prospectively assess the rectal carriage rate of third-generation cephalosporin-resistant Enterobacteriaceae (3GCREB) in non-ICU patients on hospital admission and to investigate resistance mechanisms and risk factors for carriage. METHODS: Adult patients were screened for 3GCREB carriage at six German tertiary care hospitals in 2014 using rectal swabs or stool samples. 3GCREB isolates were characterized by phenotypic and molecular methods. Each patient answered a questionnaire about potential risk factors for colonization with MDR organisms (MDROs). Univariable and multivariable risk factor analyses were performed to identify factors associated with 3GCREB carriage. RESULTS: Of 4376 patients, 416 (9.5%) were 3GCREB carriers. Escherichia coli was the predominant species (79.1%). ESBLs of the CTX-M-1 group (67.3%) and the CTX-M-9 group (16.8%) were the most frequent ß-lactamases. Five patients (0.11%) were colonized with carbapenemase-producing Enterobacteriaceae. The following risk factors were significantly associated with 3GCREB colonization in the multivariable analysis (P < 0.05): centre; previous MDRO colonization (OR = 2.12); antibiotic use within the previous 6 months (OR = 2.09); travel outside Europe (OR = 2.24); stay in a long-term care facility (OR = 1.33); and treatment of gastroesophageal reflux disease (GERD) (OR = 1.22). CONCLUSIONS: To our knowledge, this is the largest admission prevalence study of 3GCREB in Europe. The observed prevalence of 9.5% 3GCREB carriage was higher than previously reported and differed significantly among centres. In addition to previously identified risk factors, the treatment of GERD proved to be an independent risk factor for 3GCREB colonization.

Treatment resistant hyperemesis gravidarum in a patient with type 1 diabetes mellitus: neonatal withdrawal symptoms after successful antiemetic therapy with mirtazapine.

Here we present the case of a 30-year-old woman with type I diabetes mellitus, preeclampsia and treatment resistant persistent hyperemesis gravidarum in her 25th week of gestation who was successfully treated with the antidepressant mirtazapine (Remergil). Nausea and vomiting resolved within 5 days. After discharge from the hospital in 28 weeks of gestation and discontinuation of the medication on her own initiative a relapse occurred, once again with good response to mirtazapine. The drug was continued until birth. At 34 + 0 weeks a cesarean section was performed due to fetal growth restriction and deteriorating preeclampsia. During the second and fourth day postnatal age the child temporarily developed hyperarousal which could be explained by mirtazapine withdrawal.

A computer simulation study of imaging flexural inhomogeneities using plate-wave diffraction tomography.

This paper investigates the feasibility of plate-wave diffraction tomography for the reconstruction of flexural inhomogeneities in plates using the results of computer simulation studies. The numerical implementation of the fundamental reconstruction algorithm, which has recently been developed by Wang and Rose [C.H. Wang, L.R.F. Rose, Plate-wave diffraction tomography for structural health monitoring, Rev. Quant. Nondestr. Eval. 22 (2003) 1615-1622] is investigated addressing the essential effects of applying the discrete form of the Fourier diffraction theorem for solving the inverse problem as discussed by Kak and Slaney [A.C. Kak, M. Slaney, Principles of Computerized Tomographic Imaging, IEEE Press, New York, 1988] for the acoustic case, viz. diffraction limited sensitivity, influence of weak scatterer assumption, damage location and scatter field data processing in time and Fourier space as well as experimental limitations such as finite receiver length and limited views. The feasibility of the imaging technique is investigated for cylindrical inhomogeneities of various severities and relative position within the interrogation space and a normal incident interrogation configuration. The results show that plate-wave diffraction tomography enables the quantitative reconstruction of location, size and severity of plate damage with excellent sensitivity and offers the potential for detecting corrosion thinning, disbonds and delamination damage in structural integrity management applications.

The syringaldazine-oxidizing peroxidase PXP 3-4 from poplar xylem: cDNA isolation, characterization and expression.

The cell wall polymer lignin is believed to be condensed by specific cell wall-localized oxidoreductases. In many plants species, including poplar, the peroxidase-directed oxidation of the lignin analogue syringaldazine (SYR) has been localized to cells that undergo secondary wall formation, a process that includes lignification. As a first step to analyse the corresponding peroxidases. we have isolated previously two anionic isoenzymes (PXP 3-4 and PXP 5) from poplar xylem (Populus trichocarpa), which use SYR as a substrate. Here, we demonstrate that these enzymes are responsible for the visualized SYR oxidation in the developing xylem. The cDNA that corresponds to PXP 3-4 was isolated and the deduced protein was found closely related to the other SYR-oxidizing peroxidase PXP 5 (ca. 98% of identity). PXP 3-4 was expressed in a baculovirus expression system yielding high levels of active peroxidase (3 mg/l medium). The heterologously produced protein showed characteristics similar to those of the corresponding protein from poplar xylem (enzymatic properties, isoelectric point, and migration in a native gel). PXP 3-4 was expressed in the stem and in the root xylem. The data demonstrate that PXP 3-4 (and/or PXP 5) are present in differentiating xylem. supporting a function in secondary cell wall formation.

[Psychologic effects of traumatic live deliveries]. / Psychische Auswirkungen traumatisch erlebter Entbindungen.

OBJECTIVES: Traumatically experienced childbirth can lead to serious psychological disturbances postpartum. Dependent upon objective and subjective factors, some women may even develop the symptomatology of a posttraumatic stress disorder (PTSD). The aim of a pilot study was the evaluation of the frequency of traumatically experienced childbirth, of the effects of these traumatic experiences and possible risk factors for the development of PTSD. STUDY DESIGN: 976 women, who had given birth at the Bonn University Women's Hospital during 1997/1998, were retrospectively questioned regarding their experiences with pregnancy, childbirth and postpartum. 46 women, who described relevant psychological symptoms after childbirth, were personally interviewed. RESULTS: 424 women returned the completed questionnaire. 17.2% of the women reported anxiety postpartum, 9.4% depressive symptoms, 12% a mental re-experience of delivery within the first weeks postpartum, 3.8% were still suffering from these intrusions at the time of the study. In the first weeks after labor, nightmares were reported in 3.1%. In 4 cases, the full criteria for a PTSD were met. In 10 further cases, a subsyndromal form of this disorder was found. The case analysis showed that the development of PTSD symptoms was influenced by factors such as expectations, need for control, sense of shame and previous traumatic experiences. CONCLUSIONS: Psychological symptoms postpartum were reported frequently. Traumatically experienced childbirth can be responsible for specific short-term or long-term symptoms. In individual cases, a PTSD can develop after a traumatic delivery with long-term negative consequences for the health and mental condition of the mother, the mother-child-relationship and the desire for further pregnancy. In such cases, a specific psychotherapeutic treatment is always necessary.

Extraction and quantification of epibatidine in plasma.

Epibatidine was extracted from human and mouse plasma into a hexane-isopropanol mixture and back-extracted into a phosphate buffer, pH 2.5, then identified by HPLC isocratically using a CN column and quantified with ultraviolet detection at a fixed wavelength of 214 nm. The percent recovery of epibatidine from spiked plasma samples was 83.6% and the percent extraction was linear between 10 and 1,000 ng/ml. Desipramine was used as the internal standard. For spiked control samples containing 50 and 750 ng/ml, between-day precisions were 20.8 and 7.2% (RSD%), respectively; accuracy was 87.0 and 99.1%, respectively. The limit of detection was 2 ng/ml. Using this method, an intraperitoneal dose of 0.1 mg/kg of epibatidine produced mean levels of 7.3 and 37.1 ng/ml in pooled male and female plasma samples from C57BL/10 J mice, respectively. This is a simple and straightforward procedure by which plasma samples may be analyzed for epibatidine.

Hemopoietic lineage commitment decisions: in vivo evidence from a transgenic mouse model harboring micro LCR-betapro-LacZ as a transgene.

A substantial body of published data suggests activation of lineage-specific genes in multipotential hemopoietic cells before their unilineage commitment. Because the behavior and plasticity of cells isolated in vitro away from microenvironmental constraints exercised in vivo may be altered, one wonders whether similar findings can be observed in a physiologic setting in vivo. We used a transgenic mouse model harboring human micro LCR together with beta promoter sequences as a transgene to examine activation of lineage-specific programs in vivo. By using LacZ as a reporter, we had the ability to detect, quantitate, and select live cells with different levels of LacZ activation. We found strong expression of LacZ by X-gal staining in 2 lineages-erythroid and megakaryocytic. Activation in the latter was a novel finding not previously observed when similar transgenes were used. We also found activation of muLCR-betapro at low levels in progenitor cells of granulocytic-macrophagic, erythroid, or megakaryocytic lineage detected by in vitro assays, suggesting activation before commitment to a specific lineage pathway. In particular, the expression of LacZ was graded among progenitors, so that in a proportion of them activation occurred only after commitment to erythroid or megakaryocytic lineage. In addition, we found quantitative reduction in LacZ expression between fetal liver and bone marrow-derived cells, the basis of which is unclear. Collectively our data provide in vivo evidence supporting the view that lineage-specific genes are expressed in a graded fashion in pluripotential cells before their irreversible unilineage commitment. (Blood. 2000;95:1274-1282)

Quantitative trait loci and candidate gene mapping of bud set and bud flush in populus.

The genetic control of bud phenology in hybrid poplar was studied by mapping quantitative trait loci (QTL) affecting the timing of autumn bud set and spring bud flush. The founders of the mapping pedigree were collected from widely separated latitudes to maximize segregating variation for dormancy-related traits in the F(2) generation-the female Populus trichocarpa parent is from Washington State (48 degrees N) and the male P. deltoides parent is from Texas (31 degrees N). Bud set and bud flush timing were measured on the F(2) generation in a replicated clonal field trial. Using a linkage map constructed of AFLP and microsatellite markers, three QTL controlling bud set and six QTL controlling bud flush were detected. Additionally, five candidate genes believed to be involved in perception of photoperiod (PHYB1, PHYB2) or transduction of abscisic acid response signals (ABI1B, ABI1D, and ABI3) were placed on the QTL map. PHYB2 and ABI1B were found to be coincident with QTL affecting bud set and bud flush.

ABI3 affects plastid differentiation in dark-grown Arabidopsis seedlings.

The Arabidopsis ABSCISIC ACID-INSENSITIVE3 (ABI3) protein has been identified previously as a crucial regulator of late seed development. Here, we show that dark-grown abi3 plants, or abi3 plants returned to the dark after germination in the light, developed and maintained an etioplast with a prominent prolamellar body at developmental stages in which the wild type did not. Overexpression of ABI3 led to the preservation of the plastid ultrastructure that was present at the onset of darkness. These observations suggest that ABI3 plays a role in plastid differentiation pathways in vegetative tissues. Furthermore, the analysis of deetiolated (det1) abi3 double mutants revealed that DET1 and ABI3 impinge on a multitude of common processes. During seed maturation, ABI3 required DET1 to achieve its full expression. Mature det1 abi3 seeds were found to be in a highly germinative state, indicating that germination is controlled by both DET1 and ABI3. During plastid differentiation in leaves of dark-grown plants, DET1 is required for the action of ABI3 as it is during seed development. Together, the results suggest that ABI3 is at least partly regulated by light.