Detecting imipenem resistance in Acinetobacter baumannii by automated systems (BD Phoenix, Microscan WalkAway, Vitek 2); high error rates with Microscan WalkAway

<p>Abstract</p> <p>Background</p> <p>Increasing reports of carbapenem resistant <it>Acinetobacter baumannii </it>infections are of serious concern. Reliable susceptibility testing results remains a critical issue for the clinical outcome. Automated systems are increasingly used for species identification and susceptibility testing. This study was organized to evaluate the accuracies of three widely used automated susceptibility testing methods for testing the imipenem susceptibilities of <it>A. baumannii </it>isolates, by comparing to the validated test methods.</p> <p>Methods</p> <p>Selected 112 clinical isolates of <it>A. baumanii </it>collected between January 2003 and May 2006 were tested to confirm imipenem susceptibility results. Strains were tested against imipenem by the reference broth microdilution (BMD), disk diffusion (DD), Etest, BD Phoenix, MicroScan WalkAway and Vitek 2 automated systems. Data were analysed by comparing the results from each test method to those produced by the reference BMD test.</p> <p>Results</p> <p>MicroScan performed true identification of all <it>A. baumannii </it>strains while Vitek 2 unidentified one strain, Phoenix unidentified two strains and misidentified two strains. Eighty seven of the strains (78%) were resistant to imipenem by BMD. Etest, Vitek 2 and BD Phoenix produced acceptable error rates when tested against imipenem. Etest showed the best performance with only two minor errors (1.8%). Vitek 2 produced eight minor errors(7.2%). BD Phoenix produced three major errors (2.8%). DD produced two very major errors (1.8%) (slightly higher (0.3%) than the acceptable limit) and three major errors (2.7%). MicroScan showed the worst performance in susceptibility testing with unacceptable error rates; 28 very major (25%) and 50 minor errors (44.6%).</p> <p>Conclusion</p> <p>Reporting errors for <it>A. baumannii </it>against imipenem do exist in susceptibility testing systems. We suggest clinical laboratories using MicroScan system for routine use should consider using a second, independent antimicrobial susceptibility testing method to validate imipenem susceptibility. Etest, whereever available, may be used as an easy method to confirm imipenem susceptibility.</p

Identification of a TPX2-Like Microtubule-Associated Protein in Drosophila

Chromosome segregation during mitosis and meiosis relies on the spindle and the functions of numerous microtubule-associated proteins (MAPs). One of the best-studied spindle MAPs is the highly conserved TPX2, which has been reported to have characteristic intracellular dynamics and molecular activities, such as nuclear localisation in interphase, poleward movement in the metaphase spindle, microtubule nucleation, microtubule stabilisation, microtubule bundling, Aurora A kinase activation, kinesin-5 binding, and kinesin-12 recruitment. This protein has been shown to be essential for spindle formation in every cell type analysed so far. However, as yet, TPX2 homologues have not been found in the Drosophila genome. In this study, I found that the Drosophila protein Ssp1/Mei-38 has significant homology to TPX2. Sequence conservation was limited to the putative spindle microtubule-associated region of TPX2, and intriguingly, D-TPX2 (Ssp1/Mei-38) lacks Aurora A- and kinesin-5-binding domains, which are highly conserved in other animal and plant species, including many insects such as ants and bees. D-TPX2 uniformly localised to kinetochore microtubule-enriched regions of the metaphase spindle in the S2 cell line, and it had microtubule binding and bundling activities in vitro. In comparison with other systems, the contribution of D-TPX2 to cell division seems to be minor; live cell imaging of microtubules and chromosomes after RNAi knockdown identified significant delay in chromosome congression in only 18% of the cells. Thus, while this conserved spindle protein is present in Drosophila, other mechanisms may largely compensate for its spindle assembly and chromosome segregation functions

Phosphoproteomic differences in major depressive disorder postmortem brains indicate effects on synaptic function

There is still a lack in the molecular comprehension of major depressive disorder (MDD) although this condition affects approximately 10% of the world population. Protein phosphorylation is a posttranslational modification that regulates approximately one-third of the human proteins involved in a range of cellular and biological processes such as cellular signaling. Whereas phosphoproteome studies have been carried out extensively in cancer research, few such investigations have been carried out in studies of psychiatric disorders. Here, we present a comparative phosphoproteome analysis of postmortem dorsolateral prefrontal cortex tissues from 24 MDD patients and 12 control donors. Tissue extracts were analyzed using liquid chromatography mass spectrometry in a data-independent manner (LC-MSE). Our analyses resulted in the identification of 5,195 phosphopeptides, corresponding to 802 non-redundant proteins. Ninety of these proteins showed differential levels of phosphorylation in tissues from MDD subjects compared to controls, being 20 differentially phosphorylated in at least 2 peptides. The majority of these phosphorylated proteins were associated with synaptic transmission and cellular architecture not only pointing out potential biomarker candidates but mainly shedding light to the comprehension of MDD pathobiology

The elegans of spindle assembly

The Caenorhabditis elegans one-cell embryo is a powerful system in which to study microtubule organization because this large cell assembles both meiotic and mitotic spindles within the same cytoplasm over the course of 1 h in a stereotypical manner. The fertilized oocyte assembles two consecutive acentrosomal meiotic spindles that function to reduce the replicated maternal diploid set of chromosomes to a single-copy haploid set. The resulting maternal DNA then unites with the paternal DNA to form a zygotic diploid complement, around which a centrosome-based mitotic spindle forms. The early C. elegans embryo is amenable to live-cell imaging and electron tomography, permitting a detailed structural comparison of the meiotic and mitotic modes of spindle assembly

Age-Related Changes in Human Tendo Calcaneus Collagen Fibrils

Objectives: The ruptures of tendo calcaneus often occur between the age of 30 - 45 years as described by several textbooks. It is also described that some diseases and drugs are said to be responsible in the etiology; however, there are no studies related with the detailed histological structure of collagen fibrils found in the tendon in the age groups of humans. In view thereof, this study was aimed to obtain further information on the etiology and to find an answer regarding the frequency of the ruptures occurring between the age of 30 - 45 years in humans. Methods: In the study, the biopsy specimens taken from 28 patients (ages I - 68 year) who had undergone surgery due to tendo calcaneus ruptures or achilloplasty operation were examined by transmission electron microscope. All the specimens were prepared according to routine electron microscopic tissue preparation technique. The patients were divided into 7 age groups (I - 9, 10 - 19, 20 - 29, 30 - 39, 40 - 49, 50 - 59, > 60 years) and there were 4 patients in each group. The transverse diameters of collagen fibers were measured from the ultra thin sections and statistical analysis of the results were performed. The study was carried out in the electron microscopy laboratory of the Anatomy Department of Hacettepe University, Ankara, Turkey between January 2004 and September 2004. Results: The diameters of the collagen fibers were higher in the 20 - 29 year-old group compared to other groups and it showed a statistically significant difference. In patients who were in the 30 - 39 year-old group or older, the diameters of the collagen fibers were lesser than the 20 - 29 year-old group. However, an increase was observed in the collagen fibril concentration of these groups. In examination of the specimens of patients who were under 20-year-old, the diameter of the collagen fibers were less than the 20 - 29 year-old group. The electron microscopic appearance of the tissue sample of a one-year-old patient had a specific organization and in this patient, both the diameters and concentration of collagen fibers were less. Conclusion: We believe that the decrease in the diameters of collagen fibers of 30 - 45 year-old patients who are in the active period of their life, can play a role in the etiology of the frequency of tendo calcaneus ruptures similar to other etiologic factors.Wo

Non-persistent pesticides in breast milk in an agricultural area in Turkey

PubMedID: 29463139Background: Organophosphates, pythyreoids, carbamate pesticides and fungicides are heavily used in agriculture. They may have dangerous effects on newborn health especially on immune system and growth via prenatal transmission by placenta or postnatal transmission by breastfeeding. Methods: In 2015, 144 non-persistent pesticides in 64 milk samples of 32 mothers were studied by OuEChERS method in liquid chromatography/tandem mass spectrometer in neonatal Intensive Care Unit in Adana, a city in Cukurova region which is an important agricultural area in Turkey. Results: Pesticides were detected in milk samples of 11 mothers (34.3%) and 21 (32.8%) of milk samples. In five mothers, fungicides (in 5/10 samples propicanozole-PP, in 4/10 samples bromucanozole-BM), in five mothers, organophosphates (in 10/10 samples primyphosphomethyl-PPM), in one mother, both organophosphates and fungicide (in 1/2 samples PPM and in 1/2 samples buprimate) were detected. However, the estimated daily intakes (EDI) were less than acceptable daily intakes (ADI) for PPM, PP and BM, respectively. Conclusions: Although pesticides levels in human milk did not exceed the ADIs, we suggest monitoring pesticides in human breast milk especially for newborn health. © 2018, © 2018 Informa UK Limited, trading as Taylor & Francis Group.TF2014BAP9The study was funded by Cukurova University, Scientific Project Department, Project number TF2014BAP9

Identification of Candida species isolated from patients in intensive care unit and in vitro susceptibility to fluconazole for a 3-year period

Species level identification of Candida and antifungal susceptibility testing is not generally performed in routine laboratory practice. There is limited information about the distribution of Candida species and antifungal susceptibility in Turkey. In this study, we aimed at identifying Candida isolates to species level from various samples obtained from patients treated in an intensive care unit between 2002 and 2005 and to evaluate fluconazole susceptibilities of the isolates. A total of 320 Candida isolates obtained from 270 patients were identified by conventional methods and using API (Candida and/or 20C AUX) system. Antifungal susceptibility testing was performed by broth microdilution method. Candida albicans was isolated with the highest frequency (65.6%) followed by C. parapsilosis (11.3%), C. glabrata (8.8%) and C. tropicalis (7.8%). Of all the isolates, 92.9% revealed susceptibility to fluconazole. Susceptibility to fluconazole was highest for C. albicans followed by C. parapsilosis and C. glabrata. The MIC90 values for C. albicans, C. parapsilosis, C. glabrata and C. tropicalis were 1, 2, 8 and 4 µg ml -1 respectively. Fluconazole remains effective against both C. albicans and the majority of non-albicans Candida species. In this study, we determine the distribution of Candida species and evaluate the susceptibilities of the isolates, particularly for the azoles. © 2007 The Authors

Rickets with Dorfman-Chanarin syndrome

PubMedID: 17095855Background: Dorfman-Chanarin syndrome is a rare, autosomal recessive, inherited, lipid storage disease. It is characterized by nonbullous congenital ichthyosiform erythroderma, leukocyte vacuoles and variable involvement of the liver, muscles and central nervous system, due to errors of triacylglycerol metabolism. To date only 32 cases of this syndrome have been described worldwide. Aims: To report the case of a boy with Dorfman-Chanarin syndrome with rickets. Case Report: A boy of Turkish origin was born of a nonconsanguineous marriage after an uncomplicated perinatal period. On examination the patient had failure to thrive, diffuse erythroderma and ichthyosis and clinical features suggesting rickets. A light-microscopic examination of peripheral smear revealed vacuolated leukocytes typical of Jordans' anomaly. Lipid analysis showed an increase in triacylglycerol and very low density lipoprotein. A radiographic study of the wrist revealed rickets. Conclusion: The clinical progression in these patients ranges from mild to fatal. We believe that patients with ichthyosis should be evaluated bearing in mind the possibility of Dorfman-Chanarin syndrome. This may increase the number of reported Dorfman-Chanarin syndrome cases, and the pathogenesis and progression of the disease will become clearer. Copyright © 2007 S. Karger AG

Do all deliveries with elective caesarean section need paediatrician attendance

PubMedID: 22708525Aim: To determine whether paediatrician attendance to deliveries with elective caesarean section (CS) is really needed for term and also for preterm babies with 35-37 weeks gestational age. Methods: Singleton newborns ?35 gestational weeks without any identified risk factor were evaluated for resuscitation steps prospectively after CS under regional and general anaesthesia. Results: 545 infants were included in the study. 150 (27.5%) of infants needed only supplemental oxygen and 23 (4.2%) neonates needed bag and mask ventilation. None of the babies needed cardiopulmonary resuscitation (CPR) (chest compression) or endotracheal tube insertion/epinephrine administration. More infants required supplemental oxygen and bag-mask ventilation in general anaesthesia delivery group compared to spinal/epidural anaesthesia group (35.5% vs. 24.4%, p = 0.29 for oxygen and 9.2 % vs. 2.3%, p < 0.0001 for bag-mask) The need for resuscitation steps was not statistically significantly different between neonates who were born in 35-37 gestational week and neonates who were born ?38 week (p = 0.170 for supplementary oxygen, p = 0.442 for bag-mask ventilation). Conclusion: There is not increased risk for chest compression and entubation for infants ? 35 gestation weeks without antenatally identified risk factors born with elective CS either under regional or general anesthesia and only 4.2% of the babies needed bag-mask ventilation, so a health care personel who knows basic NRP may be sufficient in the clinics where it is easy to achieve an advanced skilled health care personel when needed. © 2012 Informa UK, Ltd