BACKGROUND & AIMS: :Crimean-Congo hemorrhagic fever virus (CCHFV) causes a lethal tick-borne zoonotic disease with severe clinical manifestation in humans but does not produce symptomatic disease in wild or domestic animals. The factors contributing to differential outcomes of infection between species are not yet understood. Since CCHFV is known to have tropism to kidney tissue and cattle play an important role as an amplifying host for CCHFV, in this study, we assessed in vitro cell susceptibility to CCHFV infection in immortalized and primary kidney and adrenal gland cell lines of human and bovine origin. Based on our indirect fluorescent focus assay (IFFA), we suggest a cell-to-cell CCHF viral spread process in bovine kidney cells but not in human cells. Over the course of seven days post-infection (dpi), infected bovine kidney cells are found in restricted islet-like areas. In contrast, three dpi infected human kidney or adrenal cells were noted in areas distant from one another yet progressed to up to 100% infection of the monolayer. Pronounced CCHFV replication, measured by quantitative real-time RT-PCR (qRT-PCR) of both intra- and extracellular viral RNA, was documented only in human kidney cells, supporting restrictive infection in cells of bovine origin. To further investigate the differences, lactate dehydrogenase activity and cytopathic effects were measured at different time points in all mentioned cells. In vitro assays indicated that CCHFV infection affects human and bovine kidney cells differently, where human cell lines seem to be markedly permissive. This is the initial reporting of CCHFV susceptibility and replication patterns in bovine cells and the first report to compare human and animal cell permissiveness in vitro. Further investigations will help to understand the impact of different cell types of various origins on the virus-host interaction.

背景与目标： : 克里米亚-刚果出血发热病毒 (CCHFV) 引起致命的tick传播的人畜共患病，在人类中具有严重的临床表现，但在野生或家畜中不会产生症状性疾病。尚未了解导致物种之间感染结果差异的因素。由于已知CCHFV对肾脏组织具有嗜性，并且牛作为CCHFV的扩增宿主起着重要作用，因此在这项研究中，我们评估了人类和牛起源的永生化和原发性肾脏和肾上腺细胞系对CCHFV感染的体外细胞敏感性。基于我们的间接荧光聚焦测定 (IFFA)，我们建议在牛肾细胞中而不是在人细胞中进行细胞间CCHF病毒传播过程。在感染后 (dpi) 的七天内，在胰岛样区域发现了受感染的牛肾细胞。相反，在彼此远离的区域中观察到三个dpi感染的人肾脏或肾上腺细胞，但进展到单层的100% 感染。通过定量实时rt-pcr (qRT-PCR) 对细胞内和细胞外病毒RNA进行测量，仅在人肾细胞中记录了明显的CCHFV复制，支持牛来源细胞中的限制性感染。为了进一步研究差异，在所有提到的细胞的不同时间点测量了乳酸脱氢酶活性和细胞病变作用。体外测定表明，CCHFV感染对人和牛肾细胞的影响不同，而人细胞系似乎明显允许。这是牛细胞中CCHFV敏感性和复制模式的初步报道，也是首次比较体外人类和动物细胞许可性的报道。进一步的研究将有助于了解不同来源的不同细胞类型对病毒-宿主相互作用的影响。

BACKGROUND & AIMS: :The embryonated chicken egg (ECE) provides a convenient, space-saving incubator for the cultivation of many kinds of animal viruses where the egg can be easily observed for viral replication throughout the development of the chicken embryo. Within the family Bunyaviridae, the embryonated egg has been used as a host system for many viruses such as Rift Valley fever virus and Akabane virus. The current study was conducted to determine the cultivation of Crimean-Congo hemorrhagic fever virus (CCHFV) in ECE. Four-day-old eggs were infected with CCHFV via the yolk sac route and harvested embryonic tissues and amino-allantoic fluid (AAF) that were used for virus passage and viral RNA (vRNA) detection. Quantification of vRNA copies was performed by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Our study indicated that CCHFV caused the death of the embryonated egg in a dose-dependent manner and the 50% egg infectious dose (EID50) was determined to be 6.47×10(5) copies/egg. CCHFV replicated and passaged well in the egg and high viral loads were detected both in embryonic tissue (10(9-10) copies/g) and AAF (10(7-9) copies/ml) of the embryonated egg. Thus, ECE could be used for viral cultivation and preservation, and as a potential host infection model for the study of the pathogenesis of CCHFV.

背景与目标： : 胚胎鸡蛋 (ECE) 为培养多种动物病毒提供了方便，节省空间的培养箱，在整个鸡胚发育过程中，鸡蛋可以很容易地观察到病毒复制。在Bunyaviridae家族中，胚胎卵已被用作许多病毒的宿主系统，例如裂谷发热病毒和赤羽病毒。目前的研究是为了确定欧洲经委会中克里米亚-刚果出血发热病毒 (CCHFV) 的培养。四天大的卵通过卵黄囊途径感染了CCHFV，并收获了用于病毒传代和病毒RNA (vRNA) 检测的胚胎组织和氨基尿囊液 (AAF)。通过定量逆转录聚合酶链反应 (qRT-PCR) 进行vRNA拷贝的定量。我们的研究表明，CCHFV以剂量依赖性方式导致胚胎卵死亡，50% 卵感染剂量 (EID50) 被确定为6.47 × 10(5) 个拷贝/卵。CCHFV在卵中复制和传代良好，并且在胚胎组织 (10(9-10) 拷贝/g) 和AAF (10(7-9) 拷贝/ml) 中均检测到高病毒载量。因此，ECE可用于病毒的培养和保存，并可作为研究CCHFV发病机理的潜在宿主感染模型。

BACKGROUND & AIMS: :Crimean-Congo hemorrhagic fever virus was detected in ticks removed from migratory birds in Morocco. This finding demonstrates the circulation of this virus in northwestern Africa and supports the hypothesis that the virus can be introduced into Europe by infected ticks transported from Africa by migratory birds.

背景与目标： : 在摩洛哥从候鸟身上取出的蜱中检测到克里米亚-刚果出血发热病毒。这一发现证明了这种病毒在非洲西北部的传播，并支持了这种病毒可以通过候鸟从非洲运来的感染蜱引入欧洲的假说。

BACKGROUND & AIMS: :We evaluated the feasibility of a Positive Prevention intervention adapted for youth living with HIV/AIDS (YLWH) ages 15-24 in Kinshasa, Democratic Republic of the Congo. We conducted in-depth interviews and focus group discussions with intervention facilitators and YLWH participants on the following four areas of a feasibility framework: acceptability, implementation, adaptation, and limited-efficacy. The adapted intervention was suitable, satisfying, and attractive to program facilitators and participants and able to be implemented effectively. It performed well with a new population and showed preliminary efficacy. However, we identified certain aspects of the intervention that must be addressed prior to wider implementation such as: (1) including more content on navigating marriage while living with HIV and disclosure; (2) adjusting intervention timing and session length; and (3) simplifying the more complicated content. An adapted evidencebased intervention was found to be feasible and lessons learned can be applied to YLWH in other low-resource settings.

背景与目标： : 我们评估了在刚果民主共和国金沙萨针对15-24岁的艾滋病毒/艾滋病青年 (YLWH) 采取积极预防干预措施的可行性。我们与干预促进者和YLWH参与者就可行性框架的以下四个领域进行了深入访谈和焦点小组讨论: 可接受性，实施性，适应性和有限功效。经过调整的干预措施适合，令人满意，并且对计划促进者和参与者具有吸引力，并且能够有效实施。它在新人群中表现良好，并显示出初步疗效。但是，我们确定了在更广泛实施之前必须解决的干预措施的某些方面，例如 :( 1) 包括更多有关艾滋病毒感染者和披露的婚姻导航的内容; (2) 调整干预时间和会话长度; (3) 简化更复杂的内容。发现经过调整的基于证据的干预是可行的，并且可以将经验教训应用于其他低资源环境中的YLWH。

BACKGROUND & AIMS: :The present study investigated the use of algae biorefinery waste and wastepaper in the preparation of cost-effective and eco-friendly xerogels for the removal of congo red (CR) and Fe2+. The xerogel properties such as density, swelling degree and porosity were modified by incorporating alginate extracted from the brown seaweed Cystoseira trinodis. The developed biosorbents exhibited a light and porous network structure and were characterized by a fast uptake of CR and Fe2+ and adsorption efficiency was increased at pH 6-8. The equilibrium adsorption capacity was found to be 6.20-7.28 mg CR g-1 biosorbent and 8.08-8.39 mg Fe2+ g-1 biosorbent using different xerogels. The adsorption of CR obeyed first-order kinetics, while, Fe2+ followed second-order kinetics. Intraparticle diffusion model suggested a boundary layer effect. The adsorption capacity was maximally obtained as 41.15 mg g-1 and 169.49 mg g-1 for CR and Fe2+ using wastepaper/Spirulina and wastepaper/alginate/Spirulina xerogel, respectively. Temkin isotherm fitted better to the equilibrium data of CR adsorption than Langmuir and Freundlich models. While, equilibrium data of Fe2+ exhibited a best fit to both Langmuir and Freundlich models. Additionally, the Dubinin-Radushkevich isotherm suggested that adsorption mechanism of CR or Fe2+ is predominately physisorption. Investigation of thermodynamic parameters such as ΔH° and ΔS° and ΔG° confirmed the feasibility, spontaneity, randomness and endothermic nature of the adsorption process. Electrostatic attraction, H-bonding and n-π interactions were mainly involved in the biosorption process of CR. The results of this study showed that the developed xerogels could be effectively applied for dye and heavy metal removal at low concentrations.

背景与目标： : 本研究调查了藻类生物精炼废料和废纸在制备具有成本效益且生态友好的干凝胶以去除刚果红 (CR) 和Fe2中的用途。通过掺入从棕色海藻Cystoseira trinodis中提取的藻酸盐，可以改变干凝胶的性质，例如密度，溶胀度和孔隙率。所开发的生物吸附剂表现出光和多孔网络结构，其特征在于CR和Fe2的快速吸收，并且在pH 6-8时吸附效率提高。使用不同的干凝胶，发现平衡吸附量为6.20-7.28 mg CR g-1生物吸附剂和8.08-8.39 mg Fe2 g-1生物吸附剂。CR的吸附服从一级动力学，而Fe2遵循二级动力学。体内扩散模型表明存在边界层效应。分别使用废纸/螺旋藻和废纸/藻酸盐/螺旋藻干凝胶，以41.15 mg g-1和169.49 mg g-1获得最大的吸附容量。Temkin等温线比Langmuir和Freundlich模型更适合CR吸附的平衡数据。同时，Fe2的平衡数据最适合Langmuir和Freundlich模型。此外，Dubinin-Radushkevich等温线表明CR或Fe2的吸附机理主要是物理吸附。对热力学参数 (例如 Δ h °，Δ s ° 和 Δ g °) 的研究证实了吸附过程的可行性，自发性，随机性和吸热性质。静电吸引，H键和n-π 相互作用主要参与CR的生物吸附过程。这项研究的结果表明，开发的干凝胶可以在低浓度下有效地用于去除染料和重金属。

BACKGROUND & AIMS: BACKGROUND:The new biochemical marker, signal peptide-CUB-EGF domain-containing protein 1 (SCUBE1), is secreted and cell surface glycoprotein expressed during early embryogenesis. The protein is found in platelet and endothelial cells. Crimean-Congo hemorrhagic fever (CCHF), which is caused by a tick-borne virus belonging to the Bunyaviridae family, may present with a mild clinical course or may exhibit a severe profile with potentially fatal hemorrhaging. The aim of this study was to determine the diagnostic and prognostic significance of SCUBE1 levels in CCHF. METHODS:This study was conducted with patients with CCHF. SCUBE1 levels in patients with CCHF were determined using an ELISA. RESULTS:SCUBE1 titers of CCHF patients were significantly higher compared to those of the control group (p=0.0001). SCUBE1 levels of patients with hemorrhage were significantly higher than those of patients without hemorrhage (p=0.0001). SCUBE1 values of patients who died were significantly higher than those of the survivors (p=0.012). CONCLUSIONS:SCUBE1 levels are a new biomarker that can be used in the differential diagnosis and monitoring of patients hospitalized with suspected CCHF. These levels are also significant as potential predictors of mortality.

背景与目标：

BACKGROUND & AIMS: :In the current protocol, we describe the Congo red staining method and a method for separately quantifying vascular and parenchymal amyloid deposits in brain tissue sections. Congo red staining detects amyloid deposits in brain tissue of amyloid precursor protein transgenic mice and human Alzheimer's tissue. It detects compacted amyloid in a beta-sheet secondary structure and labels amyloid in both the brain parenchyma (amyloid plaques) and blood vessels. Congophilic amyloid in blood vessels is called cerebral amyloid angiopathy (CAA). To date, analysis of CAA has largely used a severity rating scale, including both qualitative and quantitative characteristics. Here, we describe a simple method for quantifying total Congophilic staining and resolution of this staining into the parenchymal and vascular components based on morphological criteria. It is becoming increasingly important to separately quantify various components of the Alzheimer's pathology, given the advancement of amyloid-lowering therapies into clinical trials. The entire procedure for the Congo red staining can be performed at room temperature (20-25 degrees C) in a fume hood. The staining protocol should take 1 h 30 min including time for coverslipping slides. Time required for image analysis depends greatly on the number of samples being analyzed and the software being used. In our hands, 30 images can be collected per hour and quantified in a further 2 h.

背景与目标： : 在当前方案中，我们描述了刚果红染色方法以及一种用于分别定量脑组织切片中血管和实质淀粉样蛋白沉积物的方法。刚果红染色检测淀粉样蛋白前体蛋白转基因小鼠和人类阿尔茨海默氏症组织的脑组织中的淀粉样蛋白沉积。它在 β-sheet二级结构中检测压实的淀粉样蛋白，并在脑实质 (淀粉样斑块) 和血管中标记淀粉样蛋白。血管中的嗜血淀粉样蛋白被称为脑淀粉样血管病 (CAA)。迄今为止，对CAA的分析主要使用了严重性等级量表，包括定性和定量特征。在这里，我们描述了一种基于形态学标准的定量总嗜血染色和将该染色拆分为实质和血管成分的简单方法。鉴于降低淀粉样蛋白疗法已进入临床试验，单独量化阿尔茨海默氏病病理学的各个组成部分变得越来越重要。刚果红染色的整个过程可以在室温 (20-25 ℃) 在通风橱中进行。染色方案应花费1小时30分钟，包括盖玻片的时间。图像分析所需的时间在很大程度上取决于所分析的样本数量和所使用的软件。在我们的手中，每小时可以收集30张图像，并在2小时内进行量化。

BACKGROUND & AIMS: :Smooth strains of Shigella dysenteriae type 1, Shigella flexneri, Shigella boydii, and Shigella sonnei which form pigmented colonies (Pcr+) on Congo red agar were virulent in the Sereny test. Smooth variants unable to bind Congo red (Pcr-) were avirulent. Measurements of dye uptake from solution showed that S. dysenteriae type 1 bound the most dye, followed in order of uptake by S. flexneri, S. boydii, and S. sonnei. Using the salt aggregation test (SAT) to determine cell surface hydrophobicity, we found the same order of species. The SAT could not, however, detect differences in surface properties between Pcr+ and Pcr- pairs of isogenic smooth strains. Enteroinvasive Escherichia coli strains used in the study showed SAT and Congo red-binding properties which were similar to those of the S. flexneri strains. A direct correlation was found between pigment-binding ability and the presence of the large 140-megadalton plasmid in S. flexneri, enteroinvasive E. coli, and S. boydii but not in S. dysenteriae type 1 or S. sonnei strains. Congo red interacted with outer membranes and outer membrane proteins of S. dysenteriae type 1 but not with lipopolysaccharides. However, rough mutants of Shigella species deficient in lipopolysaccharides bound Congo red and formed pigmented colonies, showing that dye binding as a virulence assay may be misinterpreted in such cases. There was complete correlation of the Pcr+ phenotype with virulence in the smooth strains in this study, suggesting that Congo red binding can be utilized as a quick and reliable alternative to the Sereny test.

背景与目标： : 在刚果红琼脂上形成色素菌落 (Pcr) 的1型志贺氏菌，福氏志贺氏菌，博氏志贺氏菌和宋内志贺氏菌的光滑菌株在Sereny试验中具有毒性。无法结合刚果红 (Pcr-) 的光滑变体是无毒的。从溶液中吸收染料的测量结果表明，1型痢疾杆菌结合的染料最多，其次是S. flexneri，S. boydii和S. sonnei。使用盐聚集测试 (SAT) 来确定细胞表面的疏水性，我们发现相同的物种顺序。但是，SAT无法检测到等基因光滑菌株的Pcr和Pcr对之间的表面特性差异。研究中使用的肠侵袭性大肠杆菌菌株显示出SAT和刚果红结合特性，与S. flexneri菌株相似。在S. flexneri，肠侵袭性大肠杆菌和S. boydii中发现了色素结合能力与大140-兆达顿质粒的存在之间的直接相关性，但在S.痢疾1型或S. sonnei菌株中没有。刚果红与1型痢疾链球菌的外膜和外膜蛋白相互作用，但与脂多糖不相互作用。然而，缺乏脂多糖的志贺氏菌属物种的粗糙突变体会结合刚果红并形成色素菌落，这表明在这种情况下，染料结合作为毒力测定可能会被误解。在这项研究中，Pcr表型与光滑菌株的毒力完全相关，这表明刚果红结合可以用作Sereny测试的快速可靠的替代方法。

BACKGROUND & AIMS: BACKGROUND:Crimean-Congo hemorrhagic fever (CCHF) is a virulent tick-borne disease reported in more than 30 countries across Europe, Africa, and Asia. The disease is considered endemic in several Central Asian countries, including Tajikistan; however reports of human cases from these regions rarely reach the West. METHODS:We analyzed all historical confirmed cases of CCHF in Tajikistan, mapping these reports against geographic data to assess risk areas. In addition, comprehensive analysis was undertaken on the 2010 human CCHF cohort to demonstrate effective methodologies for diagnosing this disease in-country. RESULTS:These data show that CCHF is endemic in Tajikistan, and several large clusters have been recorded. Endemic foci of disease are localized to the southern region, with geographical factors such as altitude, monthly mean temperature, and monthly mean precipitation levels limiting establishment of tick vectors in other areas. Genomic analysis of viral RNA from a 2010 human case revealed high nucleotide homology (99%) to a strain isolated in Tajikistan in 1990. CONCLUSIONS:CCHF is an important vector-borne and nosocomial pathogen in Tajikistan. The ability to rapidly detect cases using real-time RT-PCR shortly after admission in the hospital setting allows prompt implementation of barrier nursing techniques, therefore reducing onward transmission of the virus.

背景与目标：

BACKGROUND & AIMS: Genetic analysis of wild-type Crimean-Congo hemorrhagic fever (CCHF) virus strains recovered in the European part of Russia was performed. Reverse transcriptase PCR followed by direct sequencing was used to recover partial sequences of the CCHF virus medium (M) genome segment (M segment) from four pools of Hyalomma marginatum ticks and six human patients. Phylogenetic analysis of the M-segment sequences from Russian strains revealed a close relatedness of the strains (nucleotide sequence diversity,

背景与目标： 对在俄罗斯欧洲部分回收的野生型克里米亚-刚果出血发热 (CCHF) 病毒株进行了遗传分析。使用逆转录酶PCR，然后进行直接测序，从四个边缘透明体和六个人类患者池中恢复CCHF病毒培养基 (M) 基因组片段 (M片段) 的部分序列。来自俄罗斯菌株的M段序列的系统发育分析显示了菌株的密切相关性 (核苷酸序列多样性，<或 = 5.0%)。这些毒株与来自世界其他地区的CCHF病毒有显着差异 (核苷酸序列多样性，10.3 20.4%)，这表明在俄罗斯欧洲部分回收的CCHF病毒毒株是一个不同的组。

BACKGROUND & AIMS: BACKGROUND:The two-day Primary Trauma Care (PTC) course covers the management of injured patients and takes into account resource constraints experienced in low and middle-income countries. Currently, there are no studies on the long-term impact of the course on knowledge or attitudes. The PTC course was introduced in Kongo Central Central province in the Democratic Republic of Congo (DRC) as part of a series of interventions to improve trauma care. The aim of this study was to evaluate the impact of PTC on the trauma knowledge, confidence and attitudes regarding trauma care of healthcare workers (HCWs) in the DRC over two years. METHOD:A retrospective cohort study was conducted comparing multiple-choice questionnaire (MCQ) and confidence matrix results of PTC attendees prior to the course, immediately after, and at the time of follow up at either 12, 16 or 24 months. A semi-structured questionnaire was additionally administered at follow up to explore the effect of PTC on key areas of trauma learning: skills, attitudes and relationships. RESULTS:A total of 59/80 HCWs who attended the PTC course completed follow-up questionnaires. Participants were predominantly male (42/59) with a mean age of 41.6 years. There was an increase of 4.8 in MCQ scores and 9.6 in confidence scores (p < 0.01) post-PTC. MCQ scores were maintained 24 months after the course, whereas confidence scores declined (p = 0.03). At follow-up, 36/59 participants reported that equipment was not available for procedures and 52/59 felt more could be done to better manage injured patients locally. All participants believed trauma services were important and felt that the course contributed to improving the management of trauma patients. CONCLUSIONS:This study found that knowledge gained from the PTC course was maintained over two years, although individuals felt less clinically confident. A refresher course may be appropriate within two years to improve relatively low overall knowledge scores and participants' confidence. Whilst resource constraints within the DRC may hinder trauma care development, the PTC course has equipped attendees with the knowledge, skills, confidence and attitudes to improve trauma service development in their region.

背景与目标：

BACKGROUND & AIMS: :The carboxymethyl-chitosan-montmorillonite (CMC-MMT) organic-inorganic hybrid material was applied to adsorb Pb(II) and Congo red (CR) anionic dye from aqueous solution. Molecular dynamics(MD) simulation was carried out to reveal the mechanism of CMC hybrid MMT. The results suggested that CMC has been adsorbed on the interlayer space and surface of MMT, hydrogen bonds had been formed between hydroxyl & carboxyl groups of CMC with the surface of MMT, Van der Waals interaction was also existed, while amino groups of CMC hadn't obvious interaction with the surface of MMT. Batch experiments were executed to investigate the adsorption effect of CMC-MMT was compared with Na-MMT and CMC. The results showed that CMC-MMT exhibited the higher adsorption capacity than Na-MMT and CMC, The OH and COOH or NH2 groups of CMC chains may serve as electrostatic interaction active sites and coordination sites, which improved the adsorption performance of CMC-MMT on Pb(II)，and the insertion of CMC enlarged the inter-layer space of MMT, resulting in more surface active sites in the CMC-MMT hybrid composite, which increased the adsorption of organic pollutant CR. The kinetic and isotherm studies indicated that the pseudo-second-order and the Langmuir model well described the adsorption equilibrium of Pb(II) and CR on CMC-MMT.

背景与目标： : 应用羧甲基-壳聚糖-蒙脱土 (CMC-MMT) 有机-无机杂化材料从水溶液中吸附Pb(II) 和刚果红 (CR) 阴离子染料。进行了分子动力学 (MD) 模拟，以揭示CMC混合MMT的机理。结果表明，CMC吸附在MMT的层间空间和表面，CMC的羟基和羧基与MMT表面之间形成了氢键，范德华也存在相互作用，而CMC的氨基与MMT表面没有明显的相互作用。进行了分批实验以研究CMC-MMT与Na-MMT和CMC的吸附效果。结果表明，CMC-MMT比Na-MMT和CMC具有更高的吸附能力，CMC链中的OH和COOH或NH2基团可以作为静电相互作用的活性位点和配位点，从而改善了CMC-MMT对Pb(II) 的吸附性能。CMC的插入扩大了MMT的层间空间，导致CMC-MMT杂化复合材料中存在更多的表面活性位点，从而增加了有机污染物CR的吸附。动力学和等温线研究表明，伪二阶模型和Langmuir模型很好地描述了Pb(II) 和CR在CMC-MMT上的吸附平衡。

BACKGROUND & AIMS: :IntroductionCrimean-Congo haemorrhagic fever (CCHF) is a tick-borne disease in Africa, Asia, the Balkan peninsula, the south-east of Europe and the Middle East, with mortality rates of 3-30%. Transmission can also occur through contact with infected animals or humans.AimThis observational, prospective case series aimed to investigate detectable viral genomic RNA in whole-body fluids and antibody dynamics in consecutive daily samples of patients diagnosed with CCHF until discharge from hospital.MethodsWe tested 18 patients and 824 swabs and sera with RT-PCR and 125 serum samples serologically.ResultsThe longest duration until clearance of viral RNA was 18 days from serum collection and 18, 15, 13, 19 and 17 days, respectively, from nasal, oral, genital (urethral or vaginal) and faecal swab, and urine. In seven patients, viral load decreased in serum at the same time as it increased in urine or persisted at the same logarithmic values. Despite clearance in serum, viral RNA was detected in faeces and genital swabs in two and three patients, respectively. Viral clearance from body fluids occurred earlier than from serum in eight patients on ribavirin treatment. The shortest seroconversion time was 3 days after symptom onset for IgM and IgG. Seroconversion of IgG occurred until Day 14 of symptoms.ConclusionWe report persistence of viral RNA in urine, faeces and genital swabs despite serum clearance. This may indicate a need for extending isolation precautions, re-evaluating discharge criteria and transmission risk after discharge, and considering oral swabs as a less invasive diagnostic alternative.

背景与目标： : 简介克里米亚-刚果出血发热 (CCHF) 是非洲，亚洲，巴尔干半岛，欧洲东南部和中东的tick传播疾病，死亡率为3-30%。传播也可以通过与受感染的动物或人类接触而发生。针对这种观察，前瞻性病例系列旨在调查全身体液中可检测的病毒基因组RNA和连续每日确诊为CCHF患者的抗体动态，直至出院。方法我们用rt-pcr和125血清样本血清学检测了18例患者和824份拭子和血清。结果病毒RNA清除前的最长持续时间是血清采集18天，15、13、19和17天，分别来自鼻，口腔，生殖器 (尿道或阴道) 和粪便拭子以及尿液。在7名患者中，血清中的病毒载量与尿液中的病毒载量增加的同时降低，或者以相同的对数值持续存在。尽管血清清除，但分别在两名和三名患者的粪便和生殖器拭子中检测到病毒RNA。在八名接受利巴韦林治疗的患者中，体液中的病毒清除比血清中的病毒清除早。IgM和IgG症状发作后3天是最短的血清转换时间。IgG的血清转化一直持续到症状的第14天。结论我们报告了尽管血清清除率，但尿液，粪便和生殖器拭子中病毒RNA的持久性。这可能表明需要扩大隔离预防措施，重新评估出院标准和出院后的传播风险，并考虑将口服拭子作为侵入性较小的诊断选择。

BACKGROUND & AIMS: Congo red (CR) has been reported to inhibit or enhance amyloid fibril formation by several proteins. To gain insight into the mechanism(s) for these apparently paradoxical effects, we studied as a model amyloidogenic protein, a dimeric immunoglobulin light chain variable domain. With a range of molar ratios of CR, i.e. r = [CR]/[protein dimer], we investigated the aggregation kinetics, conformation, hydrogen-deuterium exchange, and thermal stability of the protein. In addition, we used isothermal titration calorimetry to characterize the thermodynamics of CR binding to the protein. During incubation at 37 degrees C or during thermal scanning, with CR at r = 0.3, 1.3, and 4.8, protein aggregation was greatly accelerated compared with that measured in the absence of the dye. In contrast, with CR at r = 8.8, protein unfolding was favored over aggregation. The aggregates formed with CR at r = 0 or 0.3 were typical amyloid fibrils, but mixtures of amyloid fibrils and amorphous aggregates were formed at r = 1.3 and 4.8. CR decreased the apparent thermal unfolding temperature of the protein. Furthermore, CR perturbed the tertiary structure of the protein without significantly altering its secondary structure. Consistent with this result, CR also increased the rate of hydrogen-deuterium exchange by the protein. Isothermal titration calorimetry showed that CR binding to the protein was enthalpically driven, indicating that binding was mainly the result of electrostatic interactions. Overall, these results demonstrate that at low concentrations, CR binding to the protein favors a structurally perturbed, aggregation-competent species, resulting in acceleration of fibril formation. At high CR concentration, protein unfolding is favored over aggregation, and fibril formation is inhibited. Because low concentrations of CR can promote amyloid fibril formation, the therapeutic utility of this compound or its analogs to inhibit amyloidoses is questionable.

背景与目标： 据报道，刚果红 (CR) 可通过几种蛋白质抑制或增强淀粉样蛋白原纤维的形成。为了深入了解这些明显矛盾的作用的机制，我们研究了作为模型淀粉样蛋白，二聚体免疫球蛋白轻链可变结构域。在CR的摩尔比范围内，即r = [CR]/[蛋白质二聚体]，我们研究了蛋白质的聚集动力学，构象，氢-氘交换和热稳定性。此外，我们使用等温滴定量热法来表征CR与蛋白质结合的热力学。在37 ℃ 下孵育期间或在r = 0.3、1.3和4.8的情况下，与在不存在染料的情况下测得的相比，在热扫描期间，蛋白质聚集大大加速。相反，在CR为r = 8.8的情况下，蛋白质展开优于聚集。在r = 0或0.3时由CR形成的聚集体是典型的淀粉样蛋白原纤维，但是在r = 1.3和4.8时形成淀粉样蛋白原纤维和无定形聚集体的混合物。CR降低了蛋白质的表观热展开温度。此外，CR扰动了蛋白质的三级结构，而没有显着改变其二级结构。与此结果一致，CR还增加了蛋白质交换氢-氘的速率。等温滴定量热法显示CR与蛋白质的结合是由焓驱动的，表明结合主要是静电相互作用的结果。总体而言，这些结果表明，在低浓度下，CR与蛋白质的结合有利于结构扰动，具有聚集能力的物种，从而加速了原纤维的形成。在高CR浓度下，蛋白质的展开比聚集更受青睐，并且原纤维的形成受到抑制。由于低浓度的CR可以促进淀粉样蛋白原纤维的形成，因此该化合物或其类似物抑制淀粉样蛋白的治疗效用值得怀疑。

BACKGROUND & AIMS: BACKGROUND:Despite the almost universal adoption of Integrated Management of Childhood Illness (IMCI) guidelines for the diagnosis and treatment of sick children under the age of five in low- and middle-income countries, child mortality remains high in many settings. One possible explanation of the continued high mortality burden is lack of compliance with diagnostic and treatment protocols. We test this hypothesis in a sample of children with severe illness in the Democratic Republic of the Congo (DRC). METHODS:One thousand one hundred eighty under-five clinical visits were observed across a regionally representative sample of 321 facilities in the DRC. Based on a detailed list of disease symptoms observed, patients with severe febrile disease (including malaria), severe pneumonia, and severe dehydration were identified. For all three disease categories, treatments were then compared to recommended case management following IMCI guidelines. RESULTS:Out of 1180 under-five consultations observed, 332 patients (28%) had signs of severe febrile disease, 189 patients (16%) had signs of severe pneumonia, and 19 patients (2%) had signs of severe dehydration. Overall, providers gave the IMCI-recommended treatment in 42% of cases of these three severe diseases. Less than 15% of children with severe disease were recommended to receive in-patient care either in the facility they visited or in a higher-level facility. CONCLUSIONS:These results suggest that adherence to IMCI protocols for severe disease remains remarkably low in the DRC. There is a critical need to identify and implement effective approaches for improving the quality of care for severely ill children in settings with high child mortality.

背景与目标：