Comparison of metagenomic and traditional methods for diagnosis of E. coli enteric infections.

Diarrheagenic Escherichia coli, collectively known as DEC, is a leading cause of diarrhea, particularly in children in low- and middle-income countries. Diagnosing infections caused by different DEC pathotypes traditionally relies on the cultivation and identification of virulence genes, a resource-intensive and error-prone process. Here, we compared culture-based DEC identification with shotgun metagenomic sequencing of whole stool using 35 randomly drawn samples from a cohort of diarrhea-afflicted patients. Metagenomic sequencing detected the cultured isolates in 97% of samples, revealing, overall, reliable detection by this approach. Genome binning yielded high-quality E. coli metagenome-assembled genomes (MAGs) for 13 samples, and we observed that the MAG did not carry the diagnostic DEC virulence genes of the corresponding isolate in 60% of these samples. Specifically, two distinct scenarios were observed: diffusely adherent E. coli (DAEC) isolates without corresponding DAEC MAGs appeared to be relatively rare members of the microbiome, which was further corroborated by quantitative PCR (qPCR), and thus unlikely to represent the etiological agent in 3 of the 13 samples (~23%). In contrast, ETEC virulence genes were located on plasmids and largely escaped binning in associated MAGs despite being prevalent in the sample (5/13 samples or ~38%), revealing limitations of the metagenomic approach. These results provide important insights for diagnosing DEC infections and demonstrate how metagenomic methods can complement isolation efforts and PCR for pathogen identification and population abundance. IMPORTANCE: Diagnosing enteric infections based on traditional methods involving isolation and PCR can be erroneous due to isolation and other biases, e.g., the most abundant pathogen may not be recovered on isolation media. By employing shotgun metagenomics together with traditional methods on the same stool samples, we show that mixed infections caused by multiple pathogens are much more frequent than traditional methods indicate in the case of acute diarrhea. Further, in at least 8.5% of the total samples examined, the metagenomic approach reliably identified a different pathogen than the traditional approach. Therefore, our results provide a methodology to complement existing methods for enteric infection diagnostics with cutting-edge, culture-independent metagenomic techniques, and highlight the strengths and limitations of each approach.

The role of asymptomatic screening in the detection of recurrent ovarian cancer.

OBJECTIVE: To investigate the utility of asymptomatic screening, including CA-125, imaging, and pelvic exam, in the diagnosis and management of recurrent ovarian cancer. METHODS: Women with ovarian cancer whose cancer recurred after remission were categorized by first method that their provider suspected disease recurrence: CA-125, imaging, symptoms, or physical exam. Differences in clinicopathologic, primary treatment characteristics, and outcomes data including secondary cytoreductive surgery (SCS) outcome and overall survival (OS) were collected. RESULTS: 102 patients were identified at our institution from 2003 to 2015. 20 recurrences were detected by symptoms, while 62 recurrences were diagnosed first by asymptomatic rise in CA-125, 5 by pelvic exam, and 15 by imaging in the absence of known exam abnormality or rise in CA-125.Mean time to recurrence was 18.9 months, and median survival was 45.8 months. These did not vary by recurrence detection method (all p > 0.4). Patients whose disease was detected by CA-125 were less likely to undergo SCS than those detected by other means (21.7% vs. 35.0%, p = 0.007). In addition to the 5 patients whose recurrence was detected primarily by pelvic exam, an additional 10 (total n = 15) patients had an abnormal pelvic exam at time of diagnosis of recurrence. DISCUSSION: Recurrence detection method was not associated with differing rates of survival or optimal SCS, however those patients detected by CA-125 were less likely to undergo SCS. The pelvic exam was a useful tool for detecting a significant proportion of recurrences.

Maternal omega-3 polyunsaturated fatty acid supplementation on offspring hip joint conformation.

Unsaturated omega-3 fatty acids, especially docosahexaenoic acid (DHA), when fed to dogs improves cognitive and neurological development. Supplementation with omega-3 fatty acids such as DHA and eicosapentaenoic acid (EPA) has also been associated with lipid peroxidation, which in turn has been implicated in reduced body weight and altered bone formation. To assess the impact of omega-3 fatty acid supplementation on skeletal growth, diets containing three levels of DHA and EPA (0.01 and 0.01%, 0.14 and 0.12%, and 0.21 and 0.18%, respectively) were fed to bitches during gestation and lactation with puppies also supplemented through weaning. Thus, the subjects studied were the puppies supplemented with DHA and EPA through gestation and early postnatal life. The hip joint conformation of the puppies (n = 676) was recorded at adulthood using two radiographic, non-invasive evaluations. In this population, females had higher hip distraction indices (DI) than males. Males from the lower two levels of DHA and EPA supplementation had significantly smaller hip DI than all females and males from the highest DHA and EPA supplementation. In contrast, there were no diet effects on anatomical indicators of hip joint conformation and no visible arthritic changes. These data suggest that dietary supplementation of DHA and EPA during gestation and the perinatal period to weaning does not adversely influence hip joint formation of dogs.

Impact of storm runoff on Salmonella and Escherichia coli prevalence in irrigation ponds of fresh produce farms in southern Georgia.

AIMS: To examine Salmonella and Escherichia coli in storm runoff and irrigation ponds used by fresh produce growers, and compare Salmonella serovars with those found in cases of human salmonellosis. METHODS AND RESULTS: We collected water before and after rain events at two irrigation ponds on farms in southern Georgia, USA, and collected storm runoff/storm flow within the contributing watershed of each pond. Salmonella and E. coli concentrations were higher in ponds after rain events by an average of 0·46 (P < 0·01) and 0·61 (P < 0·05) log10 most probable number (MPN) per 100 ml respectively. Salmonella concentrations in storm runoff from fields and forests were not significantly higher than in ponds before rain events, but concentrations in storm flow from streams and ditches were higher by an average of 1·22 log10 MPN per 100 ml (P < 0·001). Eighteen Salmonella serovars were identified from 155 serotyped isolates, and eight serovars were shared between storm runoff/storm flow and ponds. Seven of the serovars, including five of the shared serovars, were present in cases of human illness in the study region in the same year. However, several serovars most commonly associated with human illness in the study region (e.g. Javiana, Enteritidis, and Montevideo) were not found in any water samples. CONCLUSIONS: Salmonella and E. coli concentrations in irrigation ponds were higher, on average, after rain events, but concentrations of Salmonella were low, and the ponds met FDA water quality standards based on E. coli. Some similarities and notable differences were found between Salmonella serovars in water samples and in cases of human illness. SIGNIFICANCE AND IMPACT OF THE STUDY: This study directly examined storm runoff/storm flow into irrigation ponds and quantified increases in Salmonella and E. coli following rain events, with potential implications for irrigation pond management as well as human health.

Hypocretin receptor 1 blockade produces bimodal modulation of cocaine-associated mesolimbic dopamine signaling.

RATIONALE: Cocaine addiction is a chronic psychiatric disorder characterized by pathological motivation to obtain cocaine and behavioral and neurochemical hypersensitivity to cocaine-associated cues. These features of cocaine addiction are thought to be driven by aberrant phasic dopamine signaling. We previously demonstrated that blockade of the hypocretin receptor 1 (HCRTr1) attenuates cocaine self-administration and reduces cocaine-induced enhancement of dopamine signaling. Despite this evidence, the effects of HCRTr1 blockade on endogenous phasic dopamine release are unknown. OBJECTIVE: In the current studies, we assessed whether blockade of HCRTr1 alters spontaneous and cue-evoked dopamine release in the nucleus accumbens core of freely moving rats. METHODS: We first validated the behavioral and neurochemical effects of the novel, highly selective, HCRTr1 antagonist RTIOX-276 using cocaine self-administration and fast-scan cyclic voltammetry (FSCV) in anesthetized rats. We then used FSCV in freely moving rats to examine whether RTIOX-276 impacts spontaneous and cue-evoked dopamine release. Finally, we used ex vivo slice FSCV to determine whether the effects of RTIOX-276 on dopamine signaling involve dopamine terminal adaptations. RESULTS: Doses of RTIOX-276 that attenuate the motivation for cocaine reduce spontaneous dopamine transient amplitude and cue-evoked dopamine release. Further, these doses attenuated cocaine-induced dopamine uptake inhibition at the level of dopamine terminals. CONCLUSION: Our results provide support for the standing hypothesis that HCRTr1 blockade suppresses endogenous phasic dopamine signals, likely via actions at dopamine cell bodies. These results also elucidate a second process through which HCRTr1 blockade attenuates the effects of cocaine by reducing cocaine sensitivity at dopamine terminals.

Implementation of a pharmacogenomics consult service to support the INGENIOUS trial.

Hospital systems increasingly utilize pharmacogenomic testing to inform clinical prescribing. Successful implementation efforts have been modeled at many academic centers. In contrast, this report provides insights into the formation of a pharmacogenomics consultation service at a safety-net hospital, which predominantly serves low-income, uninsured, and vulnerable populations. The report describes the INdiana GENomics Implementation: an Opportunity for the UnderServed (INGENIOUS) trial and addresses concerns of adjudication, credentialing, and funding.

Molecular Detection of Legionella spp. and their associations with Mycobacterium spp., Pseudomonas aeruginosa and amoeba hosts in a drinking water distribution system.

AIMS: This study investigated waterborne opportunistic pathogens (OPs) including potential hosts, and evaluated the use of Legionella spp. for indicating microbial water quality for OPs within a full-scale operating drinking water distribution system (DWDS). METHODS AND RESULTS: To investigate the occurrence of specific microbial pathogens within a major city DWDS we examined large volume (90 l drinking water) ultrafiltration (UF) concentrates collected from six sites between February, 2012 and June, 2013. The detection frequency and concentration estimates by qPCR were: Legionella spp. (57%/85 cell equivalent, CE l(-1) ), Mycobacterium spp. (88%/324 CE l(-1) ), Pseudomonas aeruginosa (24%/2 CE l(-1) ), Vermamoeba vermiformis (24%/2 CE l(-1) ) and Acanthamoeba spp. (42%/5 cyst equivalent, CE l(-1) ). There was no detection of the following microorganisms: human faecal indicator Bacteroides (HF183), Salmonella enterica, Campylobacter spp., Escherichia coli O157:H7, Giardia intestinalis, Cryptosporidium spp. or Naegleria fowleri. There were significant correlations between the qPCR signals of Legionella spp. and Mycobacterium spp., and their potential hosts V. vermiformis and Acanthamoeba spp. Sequencing of Legionella spp. demonstrated limited diversity, with most sequences coming from two dominant groups, of which the larger dominant group was an unidentified species. Other known species including Legionella pneumophila were detected, but at low frequency. The densities of Legionella spp. and Mycobacterium spp. were generally higher (17 and 324 folds, respectively) for distal sites relative to the entry point to the DWDS. CONCLUSIONS: Legionella spp. occurred, had significant growth and were strongly associated with free-living amoebae (FLA) and Mycobacterium spp., suggesting that Legionella spp. could provide a useful DWDS monitoring role to indicate potential conditions for non-faecal OPs. SIGNIFICANCE AND IMPACT OF THE STUDY: The results provide insight into microbial pathogen detection that may aid in the monitoring of microbial water quality within DWDS prior to customer exposures.

Prerequisites to implementing a pharmacogenomics program in a large health-care system.

Pharmacogenomics (PGx) technology is advancing rapidly; however, clinical adoption is lagging. The Indiana Institute of Personalized Medicine (IIPM) places a strong focus on translating PGx research into clinical practice. We describe what have been found to be the key requirements that must be delivered in order to ensure a successful and enduring PGx implementation within a large health-care system.

Improving end of life care: an information systems approach to reducing medical errors.

Chronic and terminally ill patients are disproportionately affected by medical errors. In addition, the elderly suffer more preventable adverse events than younger patients. Targeting system wide "error-reducing" reforms to vulnerable populations can significantly reduce the incidence and prevalence of human error in medical practice. Recent developments in health informatics, particularly the application of artificial intelligence (AI) techniques such as data mining, neural networks, and case-based reasoning (CBR), presents tremendous opportunities for mitigating error in disease diagnosis and patient management. Additionally, the ubiquity of the Internet creates the possibility of an almost ideal network for the dissemination of medical information. We explore the capacity and limitations of web-based palliative information systems (IS) to transform the delivery of care, streamline processes and improve the efficiency and appropriateness of medical treatment. As a result, medical error(s) that occur with patients dealing with severe, chronic illness and the frail elderly can be reduced.The palliative model grew out of the need for pain relief and comfort measures for patients diagnosed with cancer. Applied definitions of palliative care extend this convention, but there is no widely accepted definition. This research will discuss the development life cycle of two palliative information systems: the CONFER QOLP management information system (MIS), currently used by a community-based palliative care program in Brooklyn, New York, and the CAREN case-based reasoning prototype. CONFER is a web platform based on the idea of "eCare". CONFER uses XML (extensible mark-up language), a W3C-endorced standard mark up to define systems data. The second system, CAREN, is a CBR prototype designed for palliative care patients in the cancer trajectory. CBR is a technique, which tries to exploit the similarities of two situations and match decision-making to the best-known precedent cases. The prototype uses the opensource CASPIAN shell developed by the University of Aberystwyth, Wales and is available by anonymous FTP. We will discuss and analyze the preliminary results we have obtained using this CBR tool. Our research suggests that automated information systems can be used to improve the quality of care at the end of life and disseminate expert level 'know how' to palliative care clinicians. We will present how our CBR prototype can be successfully deployed, capable of securely transferring information using a Secure File Transfer Protocol (SFTP) and using a JAVA CBR engine.

Development of an expert system for classification of medical errors.

The 1999 report published by the Institute of Medicine (IOM) indicated that between 44,000 and 98,000 unnecessary deaths per year occurred in hospitals alone, as a result of errors committed by medical professionals in the United States. There has been considerable speculation that these figures are either overestimated or underestimated. For example, the possibility that they focus on isolated injuries rather than error, or the majority of surveyed respondents did not know what constitutes a (medical) error. These disagreements have led experts to challenge the estimates of patient harm attributable to error, as well as the methodologies used to enumerate them. Of particular concern is the process used in the identification, classification and prevention of medical errors. There have been numerous attempts to develop classifications of medical errors, and currently an abundance of taxonomies exist to describe their mechanism.In previous research, (Kopec, Kabir, Reinharth, Rothschild & Castiglione, 2003) a new taxonomy of Medical Errors was designed by expanding the IOM classification. This model and its extension can be used as a blueprint for future design, development and implementation of an expert system for classification of medical errors. Effective classification can facilitate pattern recognition, and pattern recognition will help in understanding the nature, background and abatement of medical errors. Such a system's goal will be to perform convincingly as an advisory consultant, exhibiting expertise on a par with and beyond human experts in specified domains. Despite substantial disagreement on the validity of the published figures for fatalities in hospitals in the IOM report, what is of importance is that the number of deaths caused by such errors is nonetheless alarming. The identification and classification of errors in medical care delivery is a very complex process, and this process can be facilitated and simplified by the implementation of an effective classification system.

Sleep deprivation does not affect seizure frequency during inpatient video-EEG monitoring.

OBJECTIVE: To determine whether acute sleep deprivation facilitates seizures during inpatient monitoring in a controlled protocol. METHODS: Eighty-four patients with medically refractory partial epilepsy undergoing inpatient monitoring were assigned in consecutive blocks to either sleep deprivation every other night or to normal sleep. In both groups, subjects were requested to stay awake during the day, from 6 AM to 10 PM. In the sleep deprivation group, patients also stayed awake between 10 PM and 6 AM every other night beginning with Day 2. Patients were removed from sleep deprivation if they had two or more secondarily generalized seizures within 24 hours. Patients were removed from the normal sleep group and were sleep deprived if they did not have a complex partial or secondarily generalized seizure by Day 6 of monitoring. In these patients removed from sleep deprivation or from normal sleep, data were analyzed up to and including the day of removal from the protocol. RESULTS: The sleep deprivation and normal sleep subjects did not differ in age, sex, seizure localization, or percent dosage reduction in antiepileptic drugs from baseline at days 1 to 3 of monitoring. Protocol duration was 6.5 +/- 2.4 days (mean +/- SD) for the sleep deprivation group and 5.8 +/- 2.0 days for the normal sleep group. Seizures per day for complex partial, secondarily generalized, and combined complex partial and secondarily generalized, calculated from admission until end of protocol, did not differ significantly between the two groups. CONCLUSION: Acute sleep deprivation did not affect seizure frequency during inpatient monitoring in our patients with intractable complex partial seizures with secondary generalization.

An object relations perspective on borderline personality.

One of the principal formulations of borderline personality disorder is based on object relations theory, a component of psychoanalytic theory. To remain relevant, psychoanalytic formulations must find support from empirical research. After summarizing the object relations understanding of borderline personality, the authors review studies in biological neuroscience, developmental psychology and cognitive science related to the fundamental concepts of object relations theory as it aplies to borderline pathology. This review suggests that these empirical studies support psychoanalytic formulations originally derived from clinical practice and observation.

The relationship between adolescent attitudes towards authority, self-concept, and delinquency.

Attitudes towards authority of youth and adults have been investigated in recent years in a number of Western countries. The present research focused on attitudes towards institutional authority among three groups of adolescents: nondelinquents, noninstitutionalized delinquents, and institutionalized delinquents. Relationships with self-concept were also investigated. It was found that attitudes towards parents, teachers, police, and the law were, in general, positive. Nondelinquents' attitudes were more positive than those of delinquents.

The diagnostic accuracy of mammography in the evaluation of male breast disease.

BACKGROUND: The role of mammography in the evaluation of male patients presenting with breast disease is controversial. This controversy is a function of the lack of specific data concerning the diagnostic accuracy of mammography when used in this clinical setting. The purpose of this study was to define the diagnostic accuracy of mammography in the evaluation of male breast disease. METHODS: One hundred and four prebiopsy mammograms from 100 patients with tissue diagnoses were read blindly by two independent radiologists, and placed into one of five predetermined categories: definitely malignant, possibly malignant, gynecomastia, benign mass, and normal. Radiologic/pathologic correlation was performed and the sensitivity (Sn), specificity (Sp), positive (Ppv) and negative predictive value (Npv), and accuracy (Ac) for each of the mammographic diagnostic category determined. RESULTS: The pathologic diagnoses were 12 cancers, including 1 patient with bilateral breast cancer, 70 cases of gynecomastia, 16 benign masses, and 6 normals. The accuracy data for the mammographic diagnostic categories are as follows: malignant (combined definitely and possibly malignant), Sn 92%, Sp 90%, Ppv 55%, Npv 99%, Ac 90%; and overall benignity (combined gynecomastia, benign mass, and normal), Sn 90%, Sp 92%, Ppv 99%, Npv 55%, Ac 90%. Six cancers (50%) coexisted with gynecomastia. CONCLUSIONS: Mammography can accurately distinguish between malignant and benign male breast disease. Although not a replacement for clinical examination, its routine use could substantially reduce the need for biopsy in patients whose mammograms and clinical examination suggest benign disease.

Nesidioblastosis / Nesidioblastosis

La nesidioblastosis es una causa conocida de hipoglicemia persistente en neonatos, pero las publicaciones acerca de su presencia en adultos son escasas. Se describe el caso de una mujer de 44 años de edad que se presentó con hipoglicemia grave asociada a hiperinsulinismo, en la cual no fue posible el diagnóstico de insulinoma mediante los métodos disponibles en el país ni durante la exploración quirúrgica, siendo sometida a una pancreatectomía corporocaudal. El examen histológico reveló la presencia de nesidioblastosis, dado el hallazgo de hiperplasia de células de los islotes provenientes del páncreas endocrino. El seguimiento demostró el control de la glicemia y actualmente la paciente se encuentra asistomática

The development of a psychodynamic treatment for patients with borderline personality disorder: a preliminary study of behavioral change.

This study examines the effectiveness of a modified psychodynamic treatment called Transference Focused Psychotherapy (TFP) designed specifically for patients, with borderline personality disorder (BPD). Twenty-three female patients diagnosed with DSM-IV BPD began twice-weekly TFP. Patients were assessed at baseline and at the end of 12 months of treatment with diagnostic instruments, measures of suicidality, self-injurious behavior, and measures of medical and psychiatric service utilization. Compared to the year prior to treatment, the number of patients who made suicide attempts significantly decreased, as did the medical risk and severity of medical condition following self-injurious behavior. Compared to the year prior, study patients during the treatment year had significantly fewer hospitalizations as well as number and days of psychiatric hospitalization. The dropout rate was 19.1%. This uncontrolled study is highly suggestive that this structured and manualized psychodynamic treatment modified for borderline patients shows promise for the ambulatory treatment of these patients and warrants further study.

Obstructive sleep apnea is common in medically refractory epilepsy patients.

BACKGROUND: Previous reports have documented the coexistence of obstructive sleep apnea (OSA) and epilepsy and the therapeutic effects of treatment on seizure frequency and daytime sleepiness. The authors' objective was to determine the prevalence of OSA and its association with survey items in a group of patients with medically refractory epilepsy undergoing polysomnography (PSG). METHODS: Thirty-nine candidates for epilepsy surgery without a history of OSA underwent PSG as part of a research protocol examining the relationship of interictal epileptiform discharges to sleep state. Subjects also completed questionnaires about their sleep, including validated measures of sleep-related breathing disorders (Sleep Apnea Scale of the Sleep Disorders Questionnaire [SA/SDQ]) and subjective daytime sleepiness (Epworth Sleepiness Scale [ESS]). RESULTS: One-third of subjects had OSA, defined by a respiratory disturbance index (RDI) > or = 5. Five subjects (13%) had moderate to severe OSA (RDI > 20). Subjects with OSA were more likely to be older, male, have a higher SA/SDQ score, and more likely to have seizures during sleep than those without OSA (p < 0.05). Seizure frequency per month, the number or type of antiepileptic drugs (AED) prescribed, the localization of seizures (temporal versus extratemporal), and the ESS were not statistically different between the two groups. CONCLUSIONS: In our sample, previously undiagnosed obstructive sleep apnea was common, especially among men, older subjects, and those with seizures during sleep. The impact of treating OSA on seizure frequency and daytime sleepiness in medically refractory epilepsy patients warrants further controlled study.

An Internet-based exercise as a component of an overall training program addressing medical aspects of radiation emergency management.

The use of ionizing radiation and radioactive materials continues to increase worldwide in industry, medicine, agriculture, research, electrical power generation, and nuclear weaponry. The risk of terrorism using weapons of mass destruction or simple radiological devices also has increased, leading to heightened concerns. Radiation accidents occur as a consequence of errors in transportation of radionuclides, use of radiation in medical diagnosis and therapy, industrial monitoring and sterilization procedures, and rarely, nuclear power generation. Compared to other industries, a small number of serious radiation accidents have occurred over the last six decades with recent cases in the Republic of Georgia, Peru, Japan, and Thailand. The medical, psychological, and political consequences of such accidents can be considerable. A number of programs designed to train medical responders in the techniques of radiation accident management have been developed and delivered in many countries. The low frequency of serious radiation accidents requires constant re-training, as skills are lost and medical staff turnover occurs. Not all of the training involves drills or exercises in which responders demonstrate learning or communication over the broad spectrum of medical response capabilities. Medical preparedness within the context of a total emergency response program is lacking in many parts of the world, particularly in Central and Eastern Europe and the Newly Independent States. This paper describes an effort to enhance medical preparedness in the context of a total program of international cooperation and conventions facilitated by the International Atomic Energy Agency. The paper concludes that novel application of telecommunications technology as part of a training activity in radiation accident preparedness can help address gaps in training in this field in which preparedness is essential but experience and practical field exercises are lacking.

Expanding the role of nurses in Armenia.

The dissolution of the Soviet Union and the declaration of Independence by the Republic of Armenia created the need for significant changes in the healthcare delivery system in Armenia. The desire to raise the level of health care presented challenges and opportunities for nurses within the Republic. Members of the departments of nursing at Boston City Hospital/Boston Medical Center in Boston, Massachusetts, University of Massachusetts Medical Center, Worcester, Massachusetts, and the Emergency Scientific Medical Center of Yerevan, Armenia, joined forces through a grant written by Boston University School of Medicine and sponsored by the American International Health Alliance under a cooperative agreement with the United States Agency for International Development to expand the role of nursing. This article describes the assessment, planning, implementation, and evaluation of changes to the role of nursing and the development of new roles for nurses within a hospital in the capital city of Yerevan.