Utilization of ancillary studies in the cytologic diagnosis of respiratory lesions: The papanicolaou society of cytopathology consensus recommendations for respiratory cytology

The Papanicolaou Society of Cytopathology has developed a set of guidelines for respiratory cytology including indications for sputum examination, bronchial washings and brushings, CT‐guided FNA and endobronchial ultrasound guided fine needle aspiration (EBUS‐FNA), as well as recommendations for classification and criteria, ancillary testing and post‐cytologic diagnosis management and follow‐up. All recommendation documents are based on the expertise of committee members, an extensive literature review, and feedback from presentations at national and international conferences. The guideline documents selectively present the results of these discussions. The present document summarizes recommendations for ancillary testing of cytologic samples. Ancillary testing including microbiologic, immunocytochemical, flow cytometric, and molecular testing, including next‐generation sequencing are discussed. Diagn. Cytopathol. 2016;44:1000–1009. © 2016 Wiley Periodicals, Inc.     

Mitochondria Distinguish Granule-Stored from de novo Synthesized Tumor Necrosis Factor Secretion in Human Mast Cells

Background: Mast cells are immune cells derived from hematopoietic precursors that mature in the tissue microenvironment. Mast cells are critical for allergic, immune and inflammatory processes, many of which involve tumor necrosis factor (TNF). These cells uniquely store TNF in their secretory granules. Upon stimulation, mast cells rapidly (30 min) secrete β-hexosaminidase and granule-stored TNF through degranulation, but also increase TNF mRNA and release de novo synthesized TNF 24 h later. The regulation of these two distinct pathways is poorly understood. Methods: Human LAD2 leukemic mast cells are stimulated by substance P. TNF secretion and gene expression were measured by ELISA and real-time PCR, and mitochondrial dynamics was observed in live cells under confocal microscopy. Cell energy consumption was measured in terms of oxygen consumption rate. Results: Here, we show that granule-stored TNF is preformed, and its secretion from LAD2 mast cells stimulated by substance P (1) exhibits higher energy consumption and is inhibited by the mitochondrial ATP pump blocker oligomycin, (2) shows rapid increase in intracellular calcium levels, and (3) exhibits reversible mitochondrial translocation, from a perinuclear distribution to the cell surface, as compared to de novo synthesized TNF release induced by lipopolysaccharide. This mitochondrial translocation is confirmed using primary human umbilical cord blood-derived mast cells stimulated by an allergic trigger (IgE/streptavidin). Conclusion: Our findings indicate that unique mitochondrial functions distinguish granule-stored from newly synthesized TNF release from human mast cells, thus permitting the versatile involvement of mast cells in different biological processes.     

Verordnungsverhalten von bayerischen Hausärzt*innen an der stationär-ambulanten Schnittstelle vor dem Hintergrund der Bayerischen Wirkstoffvereinbarung – Qualitative Ergebnisse der WirtMed-Studie

Bundesgesundheitsblatt - Gesundheitsforschung - Gesundheitsschutz - Zur transparenten Steuerung der Arzneimittelausgaben im Rahmen des Wirtschaftlichkeitsgebots (§ 12 Fünftes...     

TDP‐43 is consistently co‐localized with ubiquitinated inclusions in sporadic and Guam amyotrophic lateral sclerosis but not in familial amyotrophic lateral sclerosis with and without SOD1 mutations

The transactive response (TAR) DNA binding protein 43 (TDP‐43) has been recently implicated as a major component of ubiquitinated inclusions in amyotrophic lateral sclerosis (ALS, motor neuron disease: MND) and ALS‐related disorders. In this study, we examined abnormal TDP‐43 pathology in 13 sporadic ALS (SALS), six familial ALS (FALS) with and without Cu/Zn superoxide dismutase (SOD1) mutations (SOD1‐FALS and non‐SOD1‐FALS), Guam ALS, two frontotemporal lobar degeneration with MND/ALS (FTLD‐MND/ALS), one FTLD with ubiquitin‐only‐immunoreactive inclusions (FTLD‐U) and two progressive supranuclear palsy (PSP). Sections from the spinal cord were processed for immunohistochemistry using antibodies against TDP‐43, ubiquitin, p62, cystatin C, phosphorylated tau protein (P‐tau; AT8), α‐synuclein and phosphorylated neurofilament protein (P‐NF). In 12 out of 13 SALS and both Guam ALS cases ubiquitin and p62‐immunoreactive (IR) neuronal inclusions co‐localized with TDP‐43. In three out of four SOD1‐FALS and one of two non‐SOD1‐FALS cases, TDP‐43‐IR inclusions were absent despite the presence of p62 and/or ubiquitin‐IR inclusions. However, a single TDP‐43‐IR neuronal inclusion co‐localized with p62 and ubiquitin in one SOD1‐FALS (His48Gln) case. Except for one neuron in a Guam case, all TDP‐43‐IR neuronal inclusions were negative for P‐tau (AT8). TDP‐43‐IR glial inclusions and neurites were also demonstrated. The TDP‐43 is a consistent component of the ubiquitinated inclusions in SALS and Guam ALS, but TDP‐43‐IR inclusions are absent or scarce in SOD1‐FALS.     

The effects of levosimendan on the left ventricular function and protein phosphorylation in post-isschemic guinea pig hearts

The widely accepted theories for the decreased function in the stunned myocardium relate to Ca²⁺ desensitization and free radical-mediated tissue damage of the myofilaments. The aim of the present study was to examine whether the depressed contractile function and Ca²⁺ responsiveness of the stunned myocardium may be restored by a new Ca²⁺ sensitizer (levosimendan), which has been shown to improve the Ca²⁺ response of the myofilaments. The effects of levosimendan on the left ventricular function and the in vivo protein phosphorylation were examined in both the non-ischemic and the stunned myocardium. Myocardial stunning was induced in Langendorff-perfused guinea pig hearts by suspending the circulation for 8 min, followed by a 20-min reperfusion period. Perfusion of post-ischemic guinea pig hearts with levosimendan (0.03–0.48 μM, 6 min) was associated with dose- and time-dependent increases in both dP/dt_max (contractility) and dP/dt_min (speed of relaxation). When the effectiveness of levosimendan was compared in non-ischemic and post-ischemic hearts, no significant differences were noted in the relative stimulatory effects on contractility and relaxation, at any given time point (time-response curve) or concentration (dose-response curve). Perfusion of the guinea pig hearts with a high (0.3 μM) levosimendan concentration did not reveal any qualitative or quantitative difference in the phosphodiesterase inhibitory potential of the compound (elevation of tissue cyclic AMP levels and characteristics of protein phosphorylation) between the non-ischemic and the post-ischemic myocardium. However, when isoproterenol was adminstered to induce maximal in vivo phosphorylation of cardiac phosphorproteins, an attenuation of the ³²P-incorporation into troponin I was noted in the post-ischemic hearts. The decrease in isoproterenol-induced ³²P-incorporation into troponin I was associated with similar alterations in the tissue level of this protein. We conclude that the Ca²⁺ sensitizer levosimendan exerts dose- and time-dependent positive inotropic and lusitropic effects on the postischemic myocardium, lending support to the hypothesis tha Ca²⁺ desensitization of the myofibrils is involved in myocardial stunning. Received: 20 July 1998, Returned for 1. revision: 27 August 1998, 1. Revision received: 6 January 1999, Returned for 2. revision: 5 February 1999, 2. Revision received: 25 February 1999, Accepted: 3 March 1999     

A Multidisciplinary Assessment of Remote Myocardial Fibrosis After Reperfused Myocardial Infarction in Swine and Patients

In extensive nonreperfused myocardial infarction (MI), remote fibrosis has been documented. Early reperfusion by primary angioplasty represents the gold standard method to minimize the extension of the infarction. We aimed to ascertain whether fibrosis also affects remote regions in reperfused MI in swine and patients. Swine were subjected to a transient occlusion of the left anterior descending artery followed by 1-week or 1-month reperfusion. Collagen content in the remote area macroscopically, microscopically, by magnetic resonance microimaging, and at the molecular level was similar to controls. In patients with previous MI, samples from autopsies displayed a significant increase in collagen content only in the infarct region. In patients with previous MI submitted to cardiac magnetic resonance-T1 mapping, the extracellular volume fraction in remote segments was similar to that for controls. In all scenarios, the remote region did not show a significant increase of collagen content in comparison with controls.     

增加缝隙连接耦联对兔长QT综合征心律失常的影响

目的:观察增加缝隙连接耦联对兔长QT综合征(LQTS)模型室性心律失常的影响。方法:应用心肌细胞快速延迟整流钾通道(IKr)阻滞剂E-4031(0.5μmol/L)在兔左室楔型心肌块建立LQT2模型,随机分为正常组、LQT2组、100 nmol/L缝隙连接激动剂(AAP-10)干预组(AAP-100组)、500 nmol/L AAP-10干预组(AAP-500组),每组10只。采用浮置玻璃微电极法同步记录心内膜、心外膜心肌细胞跨膜动作电位及跨室壁心电图。结果:LQT2组QT间期,跨室壁复极离散度(TDR)、早期后除极(EAD)、R-on-T期前收缩和尖端扭转性室性心动过速(TdP)发生率均显著高于正常组(P<0.05)。在LQT2模拟状态下,500 nmol/L AAP显著缩短了QT间期和TDR(P<0.05),降低了EAD、R-on-T期前收缩和TdP的发生率(P<0.05)。结论:增加缝隙连接耦联能减少兔LQT2模型TDR和室性心律失常发生率。     

Detailed Dietary Assessment in Patients with Inoperable Tumors: Potential Deficits for Nutrition Care Plans

Advanced cancer often results in reduced dietary intake; however, data on actual intake at the time of diagnosis are limited. In the present study, a detailed dietary intake assessment was performed in patients with metastatic lung and upper gastrointestinal cancer, before initiation of systemic therapy. Basic demographics and performance status (PS) were recorded. Nutritional status was evaluated through anthropometry, Mini Nutritional Assessment (MNA), and 3 nonconsecutive 24-hour dietary recalls. Of the 84 patients enrolled, 61.4% were protein, energy, or protein–energy undernourished, regardless of body mass index (BMI) or MNA category. No differences in energy, macronutrients, and micronutrients intakes across BMI categories were recorded. Very low consumption of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), irrespective of energy intake, tumor site, BMI category, or PS was found. Suboptimal micronutrients intakes were recorded even in well-nourished and overweight/obese patients. Patients with adequate PS and better MNA score reported significantly higher intake of certain macro- and micronutrients (all P < 0.05). Most patients exhibited reduced dietary intake in terms of energy, macronutrient, and micronutrient. Very low EPA and DHA intake was recorded for the whole sample, whereas micronutrient suboptimal intakes were also prevalent in well-nourished or overweight patients. All the above should be taken into account during patients' nutritional care.     

An unusual presentation of Bochdalek hernia after laparoscopic surgery

Congenital diaphragmatic hernia (CDH) occurs as a result of an embryologic developmental defect during gestation. Occasionally, CDH is incidentally discovered in adulthood during radiological testing for other reasons, such as mild dyspnoea or digestive symptoms. Herein, we report a case of posterolateral diaphragmatic hernia presented in adulthood, some years after laparoscopic bariatric surgery.