Medicine by Alexandros G.Sfakianakis,Anapafseos 5 Agios Nikolaos,Crete 72100,Greece,tel :00302841026182 & 00306932607174
Facial and Scalp Swelling in the Pediatric Population With Hemophilia: A Diagnosis Pitfall.
Eplasty. 2016;16:ic15
Authors: Widjaja W, Aggarwala S, Ballieux F, Vandervord J
PMID: 27213025 [PubMed]
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Panhypopituitarism due to Absence of the Pituitary Stalk: A Rare Aetiology of Liver Cirrhosis.
Case Rep Endocrinol. 2016;2016:9071097
Authors: Gonzalez Rozas M, Hernanz Roman L, Gonzalez DG, Pérez-Castrillón JL
Abstract
Studies have established a relationship between hypothalamic-pituitary dysfunction and the onset of liver damage, which may occasionally progress to cirrhosis. Patients with hypopituitarism can develop a metabolic syndrome-like phenotype. Insulin resistance is the main pathophysiological axis of metabolic syndrome and is the causal factor in the development of nonalcoholic fatty liver disease (NAFLD). We present the case of a young patient with liver cirrhosis of unknown aetiology that was finally attributed to panhypopituitarism.
PMID: 27213061 [PubMed]
Background: Hospital governance is broadening its orientation from cost and production controls towards 'improving performance on clinical outcomes'. Given this new focus one might assume that doctors are drawn into hospital management across OECD countries. Hospital performance in terms of patient health, quality of care and efficiency outcomes is supposed to benefit from their involvement. However, international comparative evidence supporting this idea is limited. Just a few studies indicate that there may be a positive relationship between medical doctors being part of hospital boards, and overall hospital performance. More importantly, the assumed relationship between these so-called doctor managers and hospital performance has remained a 'black-box' thus far. However, there is an increasing literature on the implementation of quality management systems in hospitals and their relation with improved performance. It seems therefore fair to assume that the relation between the involvement of doctors in hospital management and improved hospital performance is partly mediated via quality management systems. The threefold aim of this paper is to 1) perform a quick scan of the current situation with regard to doctor managers in hospital management in 19 OECD countries, 2) explore the phenomenon of doctor managers in depth in 7 OECD countries, and 3) investigate whether doctor involvement in hospital management is associated with more advanced implementation of quality management systems. Methods: This study draws both on a quick scan amongst country coordinators in OECD's Health Care Quality Indicator program, and on the DUQuE project which focused on the implementation of quality management systems in European hospitals. Results: This paper reports two main findings. First, medical doctors fulfil a broad scope of managerial roles at departmental and hospital level but only partly accompanied by formal decision making responsibilities. Second, doctor managers having more formal decision making responsibilities in strategic hospital management areas is positively associated with the level of implementation of quality management systems. Conclusions: Our findings suggest that doctors are increasingly involved in hospital management in OECD countries, and that this may lead to better implemented quality management systems, when doctors take up managerial roles and are involved in strategic management decision making.
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Background: Exposure to ammonium persulfate (AP) has been reported to be the main cause of occupational asthma in hairdressers. The aim of this study is to assess how long the asthmatic response to AP can be induced after dermal sensitization in a mouse model. Methods: BALB/c mice received dermal applications of AP or dimethylsulfoxide (DMSO) (control) on days 1 and 8. They then received a single nasal instillation (challenge) of AP or saline on days 15, 22, 29, 36, 45, 60 and 90. Respiratory responsiveness to methacholine was measured 24 h after the challenge using a non-specific methacholine provocation test. Pulmonary inflammation was analysed in bronchoalveolar lavage (BAL), and total serum immunoglobulin (Ig) E, IgG1 and IgG2a were measured in serum samples. Histological analysis of lung slides was performed. Results: Mice dermally sensitized and intranasally challenged with AP showed respiratory responsiveness to methacholine as long as 45 days after initial sensitization, as well as increased percentage of neutrophils in BAL compared with the control group. At day 60, dermally sensitized mice still presented bronchial hyperresponsiveness, while the percentage of neutrophils returned to baseline levels similar to those of controls. Total serum IgE increased significantly on day 22 after dermal sensitization. Total serum IgG1 and IgG2a increased from 45 days after dermal sensitization and remained high at 90 days. Conclusions: Both respiratory responsiveness to methacholine and airway inflammation responses decrease with increasing time between sensitization and challenge. Respiratory responsiveness to methacholine tends to persist longer than inflammation.
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Background: While healthcare systems vary in their structure and available resources, it is widely recognized that medical doctors play a key role in their adaptation and performance. In this article, we examine recent government and organizational policies in two different health systems that aim to develop clinical leadership among the medical profession. Clinical leadership refers to the engagement and guiding role of physicians in health system improvement. Three dimensions are defined to conduct our analysis of engaging medical doctors in healthcare leadership: the position and status of medical doctors within the system; the broader institutional context of governmental and organizational policies to engage medical doctors in clinical leadership roles; and the main factors that may facilitate or limit achievements. Methods: Our aim in this study is exploratory. We selected two contrasting cases according to their level of institutional pluralism: one national health insurance system, Canada, and one etatist social insurance system, the Netherlands. We documented the institutional dynamics of medical doctors' engagement and leadership through secondary sources, such as government websites, key policy reports, and scholarly literature on health policies in both countries. Results: Initiatives across Canadian provinces signal that the medical profession and governments search for alternatives to involve doctors in health system improvement beyond the limitations imposed by their fundamental social contract and formal labour relations. These initiatives suggest an emerging trend toward more joint collaboration between governments and medical associations. In the Dutch system, organizational and legal attempts for integration over the past decades do not yet fit well with the ideas and interests of medical doctors. The engagement of medical doctors requires additional initiatives that are closer to their professional values and interests and that depart from an overly focus on top down performance indicators and competition. Conclusions: Different institutional contexts have different policy experiences regarding the engagement and leadership of medical doctors but seem to face similar policy challenges. Achieving alignment between soft (trust, collaboration) and hard (financial incentives) levers may require facilitative conditions at the level of the health system, like clarity and stability of broad policy orientations and openness to local experimentation.
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AimsHow future doctors might be educated and trained in order to meet the population and system needs of countries is currently being debated. Incorporation of a broad range of capabilities, encompassed within categories of management and, increasingly, leadership, form part of this discussion. The purpose of this paper is to outline a framework by which countries' progress in this area might be assessed and compared. Methods: Key databases and journals related to this area were reviewed. From relevant articles potential factors impacting on the incorporation of aspects of management and leadership within medical education and training were identified. These factors were tested via an online survey during 2013 with six members of a European Association of doctors who promote medical involvement in hospital management, including members from countries less represented in the health management literature. Results: A framework for analysing how management and leadership education is being approached within different systems of healthcare is developed and presented. Conclusions: More systematic work across a wider range of countries is needed if we are to have a better understanding of how countries within and beyond Europe are approaching and progressing the education of doctors in management and leadership.
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Background: Health policy has strengthened the demand for coordination between clinicians and managers and introduced new medical manager roles in hospitals to better connect medicine and management. These developments have created a scholarly debate of concepts and an increasing 'hybridization' of tasks and roles, yet the organizational effects are not well researched. This research introduces a multi-level governance approach and aims to explore the organizational needs of doctors using Sweden as a case study. Methods: We apply an assessment framework focusing on macro-meso levels and managerial-professional modes of hospital governance (using document analysis, secondary sources, and expert information) and expand the analysis towards the micro-level. Qualitative explorative empirical material gathered in two different studies in Swedish hospitals serves to pilot research into actor-centred perceptions of clinical management from the viewpoint of the 'managed' and the 'managing' doctors in an organization.ResultsSweden has developed a model of integrated hospital governance with complex structural coordination between medicine and management on the level of the organization. In terms of formal requirements, the professional background is less relevant for many management positions but in everyday work, medical managers are perceived primarily as colleagues and not as experts advising on managerial problems. The managers themselves seem to rely more on personal strength and medical knowledge than on management tools. Bringing doctors into management may hybridize formal roles and concepts, but it does not necessarily change the perceptions of doctors and improve managerial–professional coordination at the micro-level of the organization. Conclusion: This study brings gaps in hospital governance into view that may create organizational weaknesses and unmet management needs, thereby constraining more coordinated and integrated medical management.
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Background: The need of improving the governance of healthcare services has brought health professionals into management positions. However, both the processes and outcomes of this policy change highlight differences among the European countries. This article provides in-depth evidence that neither quantitative data nor cross-country comparisons have been able to provide regarding the influence of hybrids in the functioning of hospital organizations and impact on clinicians' autonomy and exposure to hybridization. Methods: The study was designed to witness the process of institutional change from the inside and while that process was underway. It reports a case study carried out in a public hospital in Portugal when the establishment of a clinical directorate was being negotiated. Data collection comprises semi-structured interviews with general managers and surgeons complemented with observations. Results: The clinical directorate under study illustrates a divisionalized professional bureaucracy model that combines features of professional bureaucracies and divisionalized forms. The hybrid manager is key to understand the extent to which practising clinicians are more accountable and to whom given that managerial tools of control have not been strengthened, and trust-based relations allow them to keep professional autonomy untouched. In sum, clinicians are allowed to profit from their activity and to perform autonomously from the hospital's board of directors. The advantageous conditions enjoyed by the clinical directorate intensify internal re-stratification in medicine, thus suggesting forms of divisionalized medical professionalism grounded in organizational dynamics. Conclusion: It is discussed the extent to which policy change to the governance of health organizations regarding the relationship between medicine and management is subject to specific constraints at the workplace level, thus conditioning the expected outcomes of policy setting. The study also highlights the role of hybrid managers in determining the extent to which practising professionals are more accountable to managerial criteria. The overall conclusion is that although medical and managerial values link to each other, clinicians reconfigure managerial criteria according to specific interests. Ultimately, medical autonomy and authority may be reinforced in organizational settings subject to NPM-driven reforms.
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Genome-wide identification of transcription factor (TF) binding sites is pivotal to our understanding of gene expression regulation. Although much progress has been made in the determination of potential binding regions of proteins by chromatin immunoprecipitation, this method has some inherent limitations regarding DNA enrichment efficiency and antibody necessity. Here, we report an alternative strategy for assaying in vivo TF-DNA binding in Arabidopsis (Arabidopsis thaliana) cells by tandem chromatin affinity purification (TChAP). Evaluation of TChAP using the E2Fa TF and comparison with traditional chromatin immunoprecipitation and single chromatin affinity purification illustrates the suitability of TChAP and provides a resource for exploring the E2Fa transcriptional network. Integration with transcriptome, cis-regulatory element, functional enrichment, and coexpression network analyses demonstrates the quality of the E2Fa TChAP sequencing data and validates the identification of new direct E2Fa targets. TChAP enhances both TF target mapping throughput, by circumventing issues related to antibody availability, and output, by improving DNA enrichment efficiency.
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The transcriptional coactivator ANGUSTIFOLIA3 (AN3) stimulates cell proliferation during Arabidopsis thaliana leaf development, but the molecular mechanism is largely unknown. Here, we show that inducible nuclear localization of AN3 during initial leaf growth results in differential expression of important transcriptional regulators, including GROWTH REGULATING FACTORs (GRFs). Chromatin purification further revealed the presence of AN3 at the loci of GRF5, GRF6, CYTOKININ RESPONSE FACTOR2, CONSTANS-LIKE5 (COL5), HECATE1 (HEC1), and ARABIDOPSIS RESPONSE REGULATOR4 (ARR4). Tandem affinity purification of protein complexes using AN3 as bait identified plant SWITCH/SUCROSE NONFERMENTING (SWI/SNF) chromatin remodeling complexes formed around the ATPases BRAHMA (BRM) or SPLAYED. Moreover, SWI/SNF ASSOCIATED PROTEIN 73B (SWP73B) is recruited by AN3 to the promoters of GRF5, GRF3, COL5, and ARR4, and both SWP73B and BRM occupy the HEC1 promoter. Furthermore, we show that AN3 and BRM genetically interact. The data indicate that AN3 associates with chromatin remodelers to regulate transcription. In addition, modification of SWI3C expression levels increases leaf size, underlining the importance of chromatin dynamics for growth regulation. Our results place the SWI/SNF-AN3 module as a major player at the transition from cell proliferation to cell differentiation in a developing leaf.
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Follicular Variant of Papillary Thyroid Carcinoma: Hybrid or Mixture.
Thyroid. 2016 May 23;
Authors: Daniels GH
PMID: 27214083 [PubMed - as supplied by publisher]
Letter to the Editor regarding the article "I-124 PET/CT in patients with differentiated thyroid cancer: clinical and quantitative image analysis".
Thyroid. 2016 May 23;
Authors: Kist JW, de Keizer B, Vogel WV
PMID: 27213951 [PubMed - as supplied by publisher]
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Phencyclidine Disrupts the Auditory Steady State Response in Rats.
PLoS One. 2015;10(8):e0134979
Authors: Leishman E, O'Donnell BF, Millward JB, Vohs JL, Rass O, Krishnan GP, Bolbecker AR, Morzorati SL
Abstract
The Auditory Steady-State Response (ASSR) in the electroencephalogram (EEG) is usually reduced in schizophrenia (SZ), particularly to 40 Hz stimulation. The gamma frequency ASSR deficit has been attributed to N-methyl-D-aspartate receptor (NMDAR) hypofunction. We tested whether the NMDAR antagonist, phencyclidine (PCP), produced similar ASSR deficits in rats. EEG was recorded from awake rats via intracranial electrodes overlaying the auditory cortex and at the vertex of the skull. ASSRs to click trains were recorded at 10, 20, 30, 40, 50, and 55 Hz and measured by ASSR Mean Power (MP) and Phase Locking Factor (PLF). In Experiment 1, the effect of different subcutaneous doses of PCP (1.0, 2.5 and 4.0 mg/kg) on the ASSR in 12 rats was assessed. In Experiment 2, ASSRs were compared in PCP treated rats and control rats at baseline, after acute injection (5 mg/kg), following two weeks of subchronic, continuous administration (5 mg/kg/day), and one week after drug cessation. Acute administration of PCP increased PLF and MP at frequencies of stimulation below 50 Hz, and decreased responses at higher frequencies at the auditory cortex site. Acute administration had a less pronounced effect at the vertex site, with a reduction of either PLF or MP observed at frequencies above 20 Hz. Acute effects increased in magnitude with higher doses of PCP. Consistent effects were not observed after subchronic PCP administration. These data indicate that acute administration of PCP, a NMDAR antagonist, produces an increase in ASSR synchrony and power at low frequencies of stimulation and a reduction of high frequency (> 40 Hz) ASSR activity in rats. Subchronic, continuous administration of PCP, on the other hand, has little impact on ASSRs. Thus, while ASSRs are highly sensitive to NMDAR antagonists, their translational utility as a cross-species biomarker for NMDAR hypofunction in SZ and other disorders may be dependent on dose and schedule.
PMID: 26258486 [PubMed - indexed for MEDLINE]
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Association of Concurrent fNIRS and EEG Signatures in Response to Auditory and Visual Stimuli.
Brain Topogr. 2015 Sep;28(5):710-25
Authors: Chen LC, Sandmann P, Thorne JD, Herrmann CS, Debener S
Abstract
Functional near-infrared spectroscopy (fNIRS) has been proven reliable for investigation of low-level visual processing in both infants and adults. Similar investigation of fundamental auditory processes with fNIRS, however, remains only partially complete. Here we employed a systematic three-level validation approach to investigate whether fNIRS could capture fundamental aspects of bottom-up acoustic processing. We performed a simultaneous fNIRS-EEG experiment with visual and auditory stimulation in 24 participants, which allowed the relationship between changes in neural activity and hemoglobin concentrations to be studied. In the first level, the fNIRS results showed a clear distinction between visual and auditory sensory modalities. Specifically, the results demonstrated area specificity, that is, maximal fNIRS responses in visual and auditory areas for the visual and auditory stimuli respectively, and stimulus selectivity, whereby the visual and auditory areas responded mainly toward their respective stimuli. In the second level, a stimulus-dependent modulation of the fNIRS signal was observed in the visual area, as well as a loudness modulation in the auditory area. Finally in the last level, we observed significant correlations between simultaneously-recorded visual evoked potentials and deoxygenated hemoglobin (DeoxyHb) concentration, and between late auditory evoked potentials and oxygenated hemoglobin (OxyHb) concentration. In sum, these results suggest good sensitivity of fNIRS to low-level sensory processing in both the visual and the auditory domain, and provide further evidence of the neurovascular coupling between hemoglobin concentration changes and non-invasive brain electrical activity.
PMID: 25589030 [PubMed - indexed for MEDLINE]
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Insights from ERPs into attention during recovery after cerebellar stroke: a case report.
Neurocase. 2015;21(6):721-6
Authors: Mannarelli D, Pauletti C, De Lucia MC, Currà A, Fattapposta F
Abstract
The role of the cerebellum in cognitive performance and attentional processes is a focus of research in recent years. We investigated the P300 component in a patient with a left posterior cerebellar ischemic stroke during both the acute phase and over 4 weeks of follow-up. After stroke, auditory event-related potentials showed a reduction in P3 amplitude, which appears to improve instead after 4 weeks of follow-up. These event-related potential findings could suggest a specific neural pattern of disruption in selective attention during the discrimination processes of the stimulus following a posterior cerebellar lesion. A recovery is observed in the long term.
PMID: 25372555 [PubMed - indexed for MEDLINE]
Head and neck squamous cell carcinoma (HNSCC) contributes globally to a great number of deaths and morbidity, in spite of new therapeutic strategies. There is a great need of new drugs that are significantly effective and less deleterious to the patients' general health. In this sense, phytotherapy is a tendency, with results pointing to its use as a chemo-preventive and adjuvant therapy. Therefore, the objective of this systematic review was to investigate the effects of curcumin on proliferation and survival of HNSCC.
The search was conducted on six databases: Cochrane, LILACS, EMBASE, MEDLINE, PubMed, and Web of Science. In vitro and in vivo studies that evaluated the effects of curcumin on cell viability, tumor growth, cell cycle and/or cell death pattern in HNSCC cell lines or animal models were selected.
Of the 525 initially gathered studies, 30 met the inclusion criteria. These studies demonstrated that curcumin induces cytotoxicity, apoptosis (via intrinsic pathway), and cell cycle arrest in G2/M phase in HSNCC cell lines. It also reduces tumor measurements in animal models. These events were mostly studied through MTT assay, flow cytometry, and cell cycle- and apoptosis-related proteins expression.
This systematic review demonstrated that curcumin is effective on HNSCC cell proliferation and survival, reinforcing the currently available evidence that curcumin could be an adjuvant drug in HNSCC treatment.
Difficult septal deviation cases: is it open or closed technique?
Braz J Otorhinolaryngol. 2016 Apr 29;
Authors: Şevik Eliçora S, Erdem D, Işık H, Damar M, Dinç AE
Abstract
INTRODUCTION: The aim of this study is to compare the functional aspects of open technique (OTS) and endonasal septoplasty (ENS) in "difficult septal deviation cases".
METHODS: 60 patients with severe nasal obstruction from S-shaped deformities, multiple deformities, high deviations etc. were included in the study. The OTS was used in 30 patients and the ENS was performed in 30 patients. The Nasal Obstruction Symptom Evaluation (NOSE) scale was administered preoperatively and at first month following surgery. Patients were also evaluated for pain postoperatively with Visual Analog Scale (VAS).
RESULTS: The mean NOSE score was decreased 62.5-11.0 in the OTS group and 61.3-21.33 in the ENS group. Improvement of the symptoms following the two surgical techniques is similar and no statistically significant difference was found between both techniques. Also there was no statistically significant difference in postoperative pain between the OTS and ENS groups evaluated by VAS.
CONCLUSION: ENS is as successful as the OTS in management difficult septal deviation cases. In patients with severe septal deformities type of the surgical technique should be selected according to the surgeon's experience and the patient's preference.
PMID: 27210822 [PubMed - as supplied by publisher]
Feasibility of ossicular chain reconstruction with resin cement.
Braz J Otorhinolaryngol. 2016 Apr 29;
Authors: Ribeiro FA, Tamaoki Y, Cabral GW
Abstract
INTRODUCTION: Disjunction of ossicular chain is a common finding in middle ear chronic disease. In addition to ossicular interposition, various materials have been used for reconstruction, such as ceramic prostheses, polyethylene, and titanium.
OBJECTIVE: Because of the high cost of the available options, the authors propose to reconstruct the ossicular chain with resin cement, a material typically used in dental reconstruction and fixation.
METHODS: Two anatomical parts of the temporal bones were used, creating a disjunction of the ossicular chain between the incus and staples and then reconstructing with resin cement. These reconstructions were repeated four times by three different surgeons to ensure the feasibility of the method.
RESULTS: A total of 12 reconstructions were carried out, four per surgeon. After applying the cement, it could be verified by touch that the space was filled properly by the used material. Proper articulation with motion transfer to the entire ossicular chain was also observed.
CONCLUSION: Resin cement is a suitable material in the reconstruction of ossicular chain injury, and it is inexpensive and technically simple.
PMID: 27210821 [PubMed - as supplied by publisher]
The relationship of the medial roof and the posterior wall of the maxillary sinus to the sphenoid sinus: a radiologic study.
Braz J Otorhinolaryngol. 2016 May 9;
Authors: Lee SJ
Abstract
INTRODUCTION: The medial maxillary sinus roof (MMSR) is a ridge formed by the superior margin of the maxillary sinus antrostomy. The posterior wall of the maxillary sinus is always included in operative fields.
OBJECTIVE: To perform radiologic study assessing the utility of the MMSR and the posterior wall of the maxillary sinus as a fixed landmarks for providing a safe route of entry into the sphenoid sinus.
METHODS: We reviewed 115 consecutive paranasal sinus Computed Tomographic scans (230 sides) of Korean adult patients performed from January 2014 to December 2014. Using the nasal floor as a reference point, the vertical distances to the highest MMSR, the sphenoid ostium and anterior sphenoid roof and floor were measured. Then the vertical distances from the highest MMSR to the sphenoid ostium and anterior sphenoid roof and floor were calculated. The coronal distance from the posterior wall of the maxillary sinus to the sphenoid ostium was determined.
RESULTS: The average height of the highest MMSR relative to the nasal floor was measured to be 33.83±3.40mm. The average vertical distance from the highest MMSR to the sphenoid ostium and anterior sphenoid roof and floor was 1.79±3.09mm, 12.02±2.93mm, and 6.18±2.88mm respectively. The average coronal distance from the posterior wall of the maxillary sinus to the sphenoid ostium was 0.78mm. The sphenoid ostium was behind the coronal plane of the posterior wall of the maxillary sinus most frequently in 103sides (44.4%). It was in the same coronal plane in 68 sides (29.3%) and in front of the plane in 61 sides (26.3%).
CONCLUSIONS: The MMSR and the posterior wall of the maxillary sinus can be used as a reliable landmark to localize and to enable a safe entry into the sphenoid sinus.
PMID: 27210820 [PubMed - as supplied by publisher]
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Anxiety among individuals with visual vertigo and vestibulopathy.
Disabil Rehabil. 2015;37(23):2197-202
Authors: Zur O, Schoen G, Dickstein R, Feldman J, Berner Y, Dannenbaum E, Fung J
Abstract
PURPOSE: Visual vertigo (VV) is a type of dizziness triggered by visual stimuli. Despite a high incidence, its relationship with anxiety is not well-defined or understood. This study evaluated anxiety levels in subjects with VV compared to vestibulopathic subjects without VV and healthy individuals.
METHODS: A cross-sectional study to evaluate anxiety among individuals with VV was conducted twice. The first study included 72 participants (66 to 83 years of age) from senior residential centers. The second included 31 participants from a vestibular rehabilitation program (age range 35-82 years). Study 1 also used the Activities-Specific Balance Confidence (ABC) scale and study 2 the Dizziness Handicap Inventory (DHI).
RESULTS: Subjects were classified as VV positive (27 in study 1, 10 in study 2) or vestibulopathic without VV (30 in study 1, 11 in study 2) based on Head Impulse or Dynamic Visual Acuity Tests and the Dizziness Questionnaire. The remaining were age-matched healthy controls. The outcome of each study demonstrated significantly higher anxiety levels (p = 0.0001) in the VV group compared to the other groups. The results of the ABC test demonstrated that subjects in the VV group had significantly lower self-confidence (p = 0.001) than those in the Vest and Cont groups and performed fewer balance-related activities of daily life. DHI results showed that VV group expressed higher (p < 0.001) mean scores for self-perceived feelings of dizziness and imbalance (54%), compared to the Vest (9%) and Cont groups (1%).
CONCLUSIONS: Anxiety related to VV requires special attention when assessing and managing vestibulopathy, regardless of patient age.
IMPLICATIONS FOR REHABILITATION: Anxiety is a characteristic of subjects with visual vertigo (VV) and vestibulopathy. Anxiety in subjects with VV is not related to age. VV should be considered when subjects with anxiety complain of imbalance. Anxiety and vestibulopathy are often interrelated and should be considered in diagnostic evaluations.
PMID: 25597835 [PubMed - indexed for MEDLINE]
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Periprocedural stroke presenting as isolated unilateral internuclear ophthalmoplegia.
QJM. 2015 Feb;108(2):165
Authors: Saini M, Bte Idu Jion Y
PMID: 25114281 [PubMed - indexed for MEDLINE]
Injury
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Cephalalgia
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BMC Neurology
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Seminars in Pediatric Surgery
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Journal of Trauma Nursing
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High Resolution Quantification of Crystalline Cellulose Accumulation in Arabidopsis Roots to Monitor Tissue-specific Cell Wall Modifications.
J Vis Exp. 2016;(111)
Authors: Fridman Y, Holland N, Elbaum R, Savaldi-Goldstein S
Abstract
Plant cells are surrounded by a cell wall, the composition of which determines their final size and shape. The cell wall is composed of a complex matrix containing polysaccharides that include cellulose microfibrils that form both crystalline structures and cellulose chains of amorphous organization. The orientation of the cellulose fibers and their concentrations dictate the mechanical properties of the cell. Several methods are used to determine the levels of crystalline cellulose, each bringing both advantages and limitations. Some can distinguish the proportion of crystalline regions within the total cellulose. However, they are limited to whole-organ analyses that are deficient in spatiotemporal information. Others relying on live imaging, are limited by the use of imprecise dyes. Here, we report a sensitive polarized light-based system for specific quantification of relative light retardance, representing crystalline cellulose accumulation in cross sections of Arabidopsis thaliana roots. In this method, the cellular resolution and anatomical data are maintained, enabling direct comparisons between the different tissues composing the growing root. This approach opens a new analytical dimension, shedding light on the link between cell wall composition, cellular behavior and whole-organ growth.
PMID: 27214583 [PubMed - as supplied by publisher]
Microinjection for Transgenesis and Genome Editing in Threespine Sticklebacks.
J Vis Exp. 2016;(111)
Authors: Erickson PA, Ellis NA, Miller CT
Abstract
The threespine stickleback fish has emerged as a powerful system to study the genetic basis of a wide variety of morphological, physiological, and behavioral phenotypes. The remarkably diverse phenotypes that have evolved as marine populations adapt to countless freshwater environments, combined with the ability to cross marine and freshwater forms, provide a rare vertebrate system in which genetics can be used to map genomic regions controlling evolved traits. Excellent genomic resources are now available, facilitating molecular genetic dissection of evolved changes. While mapping experiments generate lists of interesting candidate genes, functional genetic manipulations are required to test the roles of these genes. Gene regulation can be studied with transgenic reporter plasmids and BACs integrated into the genome using the Tol2 transposase system. Functions of specific candidate genes and cis-regulatory elements can be assessed by inducing targeted mutations with TALEN and CRISPR/Cas9 genome editing reagents. All methods require introducing nucleic acids into fertilized one-cell stickleback embryos, a task made challenging by the thick chorion of stickleback embryos and the relatively small and thin blastomere. Here, a detailed protocol for microinjection of nucleic acids into stickleback embryos is described for transgenic and genome editing applications to study gene expression and function, as well as techniques to assess the success of transgenesis and recover stable lines.
PMID: 27214565 [PubMed - as supplied by publisher]
Indirect Fabrication of Lattice Metals with Thin Sections Using Centrifugal Casting.
J Vis Exp. 2016;(111)
Authors: Mun J, Ju J, Thurman J
Abstract
One of the typical methods to manufacture 3D lattice metals is the direct-metal additive manufacturing (AM) process such as Selective Laser Melting (SLM) and Electron Beam Melting (EBM). In spite of its potential processing capability, the direct AM method has several disadvantages such as high cost, poor surface finish of final products, limitation in material selection, high thermal stress, and anisotropic properties of parts. We propose a cost-effective method to manufacture 3D lattice metals. The objective of this study is to provide a detailed protocol on fabrication of 3D lattice metals having a complex shape and a thin wall thickness; e.g., octet truss made of Al and Cu alloys having a unit cell length of 5 mm and a cell wall thickness of 0.5 mm. An overall experimental procedure is divided into eight sections: (a) 3D printing of sacrificial patterns (b) melt-out of support materials (c) removal of residue of support materials (d) pattern assembly (e) investment (f) burn-out of sacrificial patterns (g) centrifugal casting (h) post-processing for final products. The suggested indirect AM technique provides the potential to manufacture ultra-lightweight lattice metals; e.g., lattice structures with Al alloys. It appears that the process parameters should be properly controlled depending on materials and lattice geometry, observing the final products of octet truss metals by the indirect AM technique.
PMID: 27214495 [PubMed - as supplied by publisher]
Induction of Experimental Autoimmune Encephalomyelitis in Mice and Evaluation of the Disease-dependent Distribution of Immune Cells in Various Tissues.
J Vis Exp. 2016;(111)
Authors: Barthelmes J, Tafferner N, Kurz J, de Bruin N, Parnham MJ, Geisslinger G, Schiffmann S
Abstract
Multiple sclerosis is presumed to be an inflammatory autoimmune disease, which is characterized by lesion formation in the central nervous system (CNS) resulting in cognitive and motor impairment. Experimental autoimmune encephalomyelitis (EAE) is a useful animal model of MS, because it is also characterized by lesion formation in the CNS, motor impairment and is also driven by autoimmune and inflammatory reactions. One of the EAE models is induced with a peptide derived from the myelin oligodendrocyte protein (MOG)35-55 in mice. The EAE mice develop a progressive disease course. This course is divided into three phases: the preclinical phase (day 0 - 9), the disease onset (day 10 - 11) and the acute phase (day 12 - 14). MS and EAE are induced by autoreactive T cells that infiltrate the CNS. These T cells secrete chemokines and cytokines which lead to the recruitment of further immune cells. Therefore, the immune cell distribution in the spinal cord during the three disease phases was investigated. To highlight the time point of the disease at which the activation/proliferation/accumulation of T cells, B cells and monocytes starts, the immune cell distribution in lymph nodes, spleen and blood was also assessed. Furthermore, the levels of several cytokines (IL-1β, IL-6, IL-23, TNFα, IFNγ) in the three disease phases were determined, to gain insight into the inflammatory processes of the disease. In conclusion, the data provide an overview of the functional profile of immune cells during EAE pathology.
PMID: 27214391 [PubMed - as supplied by publisher]
Generation of ESC-derived Mouse Airway Epithelial Cells Using Decellularized Lung Scaffolds.
J Vis Exp. 2016;(111)
Authors: Shojaie S, Lee J, Wang J, Ackerley C, Post M
Abstract
Lung lineage differentiation requires integration of complex environmental cues that include growth factor signaling, cell-cell interactions and cell-matrix interactions. Due to this complexity, recapitulation of lung development in vitro to promote differentiation of stem cells to lung epithelial cells has been challenging. In this protocol, decellularized lung scaffolds are used to mimic the 3-dimensional environment of the lung and generate stem cell-derived airway epithelial cells. Mouse embryonic stem cell are first differentiated to the endoderm lineage using an embryoid body (EB) culture method with activin A. Endoderm cells are then seeded onto decellularized scaffolds and cultured at air-liquid interface for up to 21 days. This technique promotes differentiation of seeded cells to functional airway epithelial cells (ciliated cells, club cells, and basal cells) without additional growth factor supplementation. This culture setup is defined, serum-free, inexpensive, and reproducible. Although there is limited contamination from non-lung endoderm lineages in culture, this protocol only generates airway epithelial populations and does not give rise to alveolar epithelial cells. Airway epithelia generated with this protocol can be used to study cell-matrix interactions during lung organogenesis and for disease modeling or drug-discovery platforms of airway-related pathologies such as cystic fibrosis.
PMID: 27214388 [PubMed - as supplied by publisher]
A Microfluidic Model of Biomimetically Breathing Pulmonary Acinar Airways.
J Vis Exp. 2016;(111)
Authors: Fishler R, Sznitman J
Abstract
Quantifying respiratory flow characteristics in the pulmonary acinar depths and how they influence inhaled aerosol transport is critical towards optimizing drug inhalation techniques as well as predicting deposition patterns of potentially toxic airborne particles in the pulmonary alveoli. Here, soft-lithography techniques are used to fabricate complex acinar-like airway structures at the truthful anatomical length-scales that reproduce physiological acinar flow phenomena in an optically accessible system. The microfluidic device features 5 generations of bifurcating alveolated ducts with periodically expanding and contracting walls. Wall actuation is achieved by altering the pressure inside water-filled chambers surrounding the thin PDMS acinar channel walls both from the sides and the top of the device. In contrast to common multilayer microfluidic devices, where the stacking of several PDMS molds is required, a simple method is presented to fabricate the top chamber by embedding the barrel section of a syringe into the PDMS mold. This novel microfluidic setup delivers physiological breathing motions which in turn give rise to characteristic acinar air-flows. In the current study, micro particle image velocimetry (µPIV) with liquid suspended particles was used to quantify such air flows based on hydrodynamic similarity matching. The good agreement between µPIV results and expected acinar flow phenomena suggest that the microfluidic platform may serve in the near future as an attractive in vitro tool to investigate directly airborne representative particle transport and deposition in the acinar regions of the lungs.
PMID: 27214269 [PubMed - as supplied by publisher]
Substantiating Appropriate Motion Capture Techniques for the Assessment of Nordic Walking Gait and Posture in Older Adults.
J Vis Exp. 2016;(111)
Authors: Dalton CM, Nantel J
Abstract
Nordic walking (NW) has become a safe and simple form of exercise in recent years, and in studying this gait pattern, various data collection techniques have been employed, each with positives and negatives. The aim was to determine the effect of NW on older adult gait and posture and to determine optimal use of different data collection systems in both short and long duration analysis. Gait and posture during NW and normal walking were assessed in 17 healthy older adults (age: 69 ± 7.3). Participants performed two trials of 6 Minute Walk Tests (6MWT) (1 with poles (WP) and 1 without poles (NP)) and 6 trials of a 5m walk (3 WP and 3 NP). Motion was recorded using two systems, a 6-sensor accelerometry system and an 8-camera 3-dimensional motion capture system, in order to quantify spatial-temporal, kinematic, and kinetic parameters. With both systems, participants demonstrated increased stride length and double support and decreased gait speed and cadence WP compared to NP (p <0.05). Also, with motion capture, larger single support time was found WP (p <0.05). With 3-D capture, smaller hip power generation and moments of force were found at heel contact and pre-swing as well as smaller knee power absorption at heel contact, pre-swing, and terminal swing WP compared to NP, when assessed over one cycle (p <0.05). Also, WP yielded smaller moments of force at heel contact and terminal swing along with larger moments at mid-stance of a gait cycle (p <0.05). No changes were found for posture. NW seems appropriate for promoting a normal gait pattern in older adults. Three-dimensional motion capture should primarily be used during short duration gait analysis (i.e. single gait cycle), while accelerometry systems should be primarily employed in instances requiring longer duration analysis such as during the 6MWT.
PMID: 27214263 [PubMed - as supplied by publisher]
Multicolor Fluorescence Detection for Droplet Microfluidics Using Optical Fibers.
J Vis Exp. 2016;(111)
Authors: Cole RH, Gartner ZJ, Abate AR
Abstract
Fluorescence assays are the most common readouts used in droplet microfluidics due to their bright signals and fast time response. Applications such as multiplex assays, enzyme evolution, and molecular biology enhanced cell sorting require the detection of two or more colors of fluorescence. Standard multicolor detection systems that couple free space lasers to epifluorescence microscopes are bulky, expensive, and difficult to maintain. In this paper, we describe a scheme to perform multicolor detection by exciting discrete regions of a microfluidic channel with lasers coupled to optical fibers. Emitted light is collected by an optical fiber coupled to a single photodetector. Because the excitation occurs at different spatial locations, the identity of emitted light can be encoded as a temporal shift, eliminating the need for more complicated light filtering schemes. The system has been used to detect droplet populations containing four unique combinations of dyes and to detect sub-nanomolar concentrations of fluorescein.
PMID: 27214249 [PubMed - as supplied by publisher]
Targeting Biofilm Associated Staphylococcus aureus Using Resazurin Based Drug-susceptibility Assay.
J Vis Exp. 2016;(111)
Authors: Dalecki AG, Crawford CL, Wolschendorf F
Abstract
Most pathogenic bacteria are able to form biofilms during infection, but due to the difficulty of manipulating and assessing biofilms, the vast majority of laboratory work is conducted with planktonic cells. Here, we describe a peg plate biofilm assay as performed with Staphylococcus aureus. Bacterial biofilms are grown on pegs attached to a 96-well microtiter plate lid, washed through gentle submersion in buffer, and placed in a drug challenge plate. After subsequent incubation they are again washed and moved to a final recovery plate, in which the fluorescent dye resazurin serves as a viability indicator. This assay offers greatly increased ease-of-use, reliability, and reproducibility, as well as a wealth of data when conducted as a kinetic read. Moreover, this assay can be adapted to a medium-throughput drug screening approach by which an endpoint fluorescent readout is taken instead, offering a path for drug discovery efforts.
PMID: 27214174 [PubMed - as supplied by publisher]
Preparation and Analysis of In Vitro Three Dimensional Breast Carcinoma Surrogates.
J Vis Exp. 2016;(111)
Authors: Goliwas KF, Miller LM, Marshall LE, Berry JL, Frost AR
Abstract
Three dimensional (3D) culture is a more physiologically relevant method to model cell behavior in vitro than two dimensional culture. Carcinomas, including breast carcinomas, are complex 3D tissues composed of cancer epithelial cells and stromal components, including fibroblasts and extracellular matrix (ECM). Yet most in vitro models of breast carcinoma consist only of cancer epithelial cells, omitting the stroma and, therefore, the 3D architecture of a tumor in vivo. Appropriate 3D modeling of carcinoma is important for accurate understanding of tumor biology, behavior, and response to therapy. However, the duration of culture and volume of 3D models is limited by the availability of oxygen and nutrients within the culture. Herein, we demonstrate a method in which breast carcinoma epithelial cells and stromal fibroblasts are incorporated into ECM to generate a 3D breast cancer surrogate that includes stroma and can be cultured as a solid 3D structure or by using a perfusion bioreactor system to deliver oxygen and nutrients. Following setup and an initial growth period, surrogates can be used for preclinical drug testing. Alternatively, the cellular and matrix components of the surrogate can be modified to address a variety of biological questions. After culture, surrogates are fixed and processed to paraffin, in a manner similar to the handling of clinical breast carcinoma specimens, for evaluation of parameters of interest. The evaluation of one such parameter, the density of cells present, is explained, where ImageJ and CellProfiler image analysis software systems are applied to photomicrographs of histologic sections of surrogates to quantify the number of nucleated cells per area. This can be used as an indicator of the change in cell number over time or the change in cell number resulting from varying growth conditions and treatments.
PMID: 27214165 [PubMed - as supplied by publisher]
Neuronavigation-guided Repetitive Transcranial Magnetic Stimulation for Aphasia.
J Vis Exp. 2016;(111)
Authors: Kim WJ, Hahn SJ, Kim WS, Paik NJ
Abstract
Repetitive transcranial magnetic stimulation (rTMS) is widely used for several neurological conditions, as it has gained acknowledgement for its potential therapeutic effects. Brain excitability is non-invasively modulated by rTMS, and rTMS to the language areas has proved its potential effects on treatment of aphasia. In our protocol, we aim to artificially induce virtual aphasia in healthy subjects by inhibiting Brodmann area 44 and 45 using neuronavigational TMS (nTMS), and F3 of the International 10-20 EEG system for conventional TMS (cTMS). To measure the degree of aphasia, changes in reaction time to a picture naming task pre- and post-stimulation are measured and compare the delay in reaction time between nTMS and cTMS. Accuracy of the two TMS stimulation methods is compared by averaging the Talairach coordinates of the target and the actual stimulation. Consistency of stimulation is demonstrated by the error range from the target. The purpose of this study is to demonstrate use of nTMS and to describe the benefits and limitations of the nTMS compared to those of cTMS.
PMID: 27214154 [PubMed - as supplied by publisher]
Characterization, Quantification and Compound-specific Isotopic Analysis of Pyrogenic Carbon Using Benzene Polycarboxylic Acids (BPCA).
J Vis Exp. 2016;(111)
Authors: Wiedemeier DB, Lang SQ, Gierga M, Abiven S, Bernasconi SM, Früh-Green GL, Hajdas I, Hanke UM, Hilf MD, McIntyre CP, Scheider MP, Smittenberg RH, Wacker L, Wiesenberg GL, Schmidt MW
Abstract
Fire-derived, pyrogenic carbon (PyC), sometimes called black carbon (BC), is the carbonaceous solid residue of biomass and fossil fuel combustion, such as char and soot. PyC is ubiquitous in the environment due to its long persistence, and its abundance might even increase with the projected increase in global wildfire activity and the continued burning of fossil fuel. PyC is also increasingly produced from the industrial pyrolysis of organic wastes, which yields charred soil amendments (biochar). Moreover, the emergence of nanotechnology may also result in the release of PyC-like compounds to the environment. It is thus a high priority to reliably detect, characterize and quantify these charred materials in order to investigate their environmental properties and to understand their role in the carbon cycle. Here, we present the benzene polycarboxylic acid (BPCA) method, which allows the simultaneous assessment of PyC's characteristics, quantity and isotopic composition ((13)C and (14)C) on a molecular level. The method is applicable to a very wide range of environmental sample materials and detects PyC over a broad range of the combustion continuum, i.e., it is sensitive to slightly charred biomass as well as high temperature chars and soot. The BPCA protocol presented here is simple to employ, highly reproducible, as well as easily extendable and modifiable to specific requirements. It thus provides a versatile tool for the investigation of PyC in various disciplines, ranging from archeology and environmental forensics to biochar and carbon cycling research.
PMID: 27214064 [PubMed - as supplied by publisher]
Development of an Ethanol-induced Fibrotic Liver Model in Zebrafish to Study Progenitor Cell-mediated Hepatocyte Regeneration.
J Vis Exp. 2016;(111)
Authors: Huang M, Xu J, Shin CH
Abstract
Sustained liver fibrosis with continuation of extracellular matrix (ECM) protein build-up results in the loss of cellular competency of the liver, leading to cirrhosis with hepatocellular dysfunction. Among multiple hepatic insults, alcohol abuse can lead to significant health problems including liver failure and hepatocellular carcinoma. Nonetheless, the identity of endogenous cellular sources that regenerate hepatocytes in response to alcohol has not been properly investigated. Moreover, few studies have effectively modeled hepatocyte regeneration upon alcohol-induced injury. We recently reported on establishing an ethanol (EtOH)-induced fibrotic liver model in zebrafish in which hepatic progenitor cells (HPCs) gave rise to hepatocytes upon near-complete hepatocyte loss in the presence of fibrogenic stimulus. Furthermore, through chemical screens using this model, we identified multiple small molecules that enhance hepatocyte regeneration. Here we describe in detail the procedures to develop an EtOH-induced fibrotic liver model and to perform chemical screens using this model in zebrafish. This protocol will be a critical tool to delineate the molecular and cellular mechanisms of how hepatocyte regenerates in the fibrotic liver. Furthermore, these methods will facilitate potential discovery of novel therapeutic strategies for chronic liver disease in vivo.
PMID: 27214059 [PubMed - as supplied by publisher]
Construction of Cell-based Neurotransmitter Fluorescent Engineered Reporters (CNiFERs) for Optical Detection of Neurotransmitters In Vivo.
J Vis Exp. 2016;(111)
Authors: Lacin E, Muller A, Fernando M, Kleinfeld D, Slesinger PA
Abstract
Cell-based neurotransmitter fluorescent engineered reporters (CNiFERs) provide a new tool for neuroscientists to optically detect the release of neurotransmitters in the brain in vivo. A specific CNiFER is created from a human embryonic kidney cell that stably expresses a specific G protein-coupled receptor, which couples to Gq/11 G proteins, and a FRET-based Ca(2+)-detector, TN-XXL. Activation of the receptor leads to an increase in the FRET signal. CNiFERs have nM sensitivity and a temporal response of seconds because a CNiFER clone utilizes the native receptor for a particular neurotransmitter, e.g., D2R for dopamine. CNiFERs are directly implanted into the brain, enabling them to sense neurotransmitter release with a spatial resolution of less than one hundred µm, making them ideal to measure volume transmission in vivo. CNiFERs can also be used to screen other drugs for potential cross-reactivity in vivo. We recently expanded the family of CNiFERs to include GPCRs that couple to Gi/o G proteins. CNiFERs are available for detecting acetylcholine (ACh), dopamine (DA) and norepinephrine (NE). Given that any GPCR can be used to create a novel CNiFER and that there are approximately 800 GPCRs in the human genome, we describe here the general procedure to design, realize, and test any type of CNiFER.
PMID: 27214050 [PubMed - as supplied by publisher]
