Decoding the distribution of glycan receptors for human-adapted influenza A viruses in ferret respiratory tract

Ferrets are widely used as animal models for studying influenza A viral pathogenesis and transmissibility. Human-adapted influenza A viruses primarily target the upper respiratory tract in humans (infection of the lower respiratory tract is observed less frequently), while in ferrets, upon intranasal inoculation both upper and lower respiratory tract are targeted. Viral tropism is governed by distribution of complex sialylated glycan receptors in various cells/tissues of the host that are specifically recognized by influenza A virus hemagglutinin (HA), a glycoprotein on viral surface. It is generally known that upper respiratory tract of humans and ferrets predominantly express α2→6 sialylated glycan receptors. However much less is known about the fine structure of these glycan receptors and their distribution in different regions of the ferret respiratory tract. In this study, we characterize distribution of glycan receptors going beyond terminal sialic acid linkage in the cranial and caudal regions of the ferret trachea (upper respiratory tract) and lung hilar region (lower respiratory tract) by multiplexing use of various plant lectins and human-adapted HAs to stain these tissue sections. Our findings show that the sialylated glycan receptors recognized by human-adapted HAs are predominantly distributed in submucosal gland of lung hilar region as a part of O-linked glycans. Our study has implications in understanding influenza A viral pathogenesis in ferrets and also in employing ferrets as animal models for developing therapeutic strategies against influenza.     

268例难治性下呼吸道感染的细菌学分析

通过本组２６８ 例难治性呼吸道感染病人标本培养结果分析,阳性２４６ 例占９１．８ ％ ,有１５４ 例检出厌氧菌,占５７ ％ ,１０４ 例检出需氧菌,占３９ ％ ,混合感染１３３ 例,占５０ ％ 。试验结果证实,厌氧菌感染在难治性下呼吸道感染病例中占了很大比例,这一结果为难治性下呼吸道感染的治疗提供了有价值的参考数据。     

Acute effects of parainfluenza virus on epithelial electrolyte transport

Parainfluenza viruses are important causes of respiratory disease in both children and adults. In particular, they are the major cause of the serious childhood illness croup (laryngotracheobronchitis). The infections produced by parainfluenza viruses are associated with the accumulation of ions and fluid in the respiratory tract. It is not known, however, whether this accumulation is because of a direct effect of the viruses on ion and fluid transport by the respiratory epithelium. Here we show that a model parainfluenza virus (the Sendai virus), in concentrations observed during respiratory infections, activates Cl- secretion and inhibits Na+ absorption across the tracheal epithelium. It does so by binding to a neuraminidase-insensitive glycolipid, possibly asialo-GM1, triggering the release of ATP, which then acts in an autocrine fashion on apical P2Y receptors to produce the observed changes in ion transport. These findings indicate that fluid accumulation in the respiratory tract associated with parainfluenza virus infection is attributable, at least in part, to direct effects of the virus on ion transport by the respiratory epithelium.     

肺病患者呼吸道曲霉随机扩增多态性DNA基因型分析

目的：了解肺病患者呼吸道曲霉基因分型状况,寻找肺部曲霉感染的可能来源,为曲霉感染的防治提供参考。方法：选择有呼吸道症状的患者925例作受试对象,收集其鼻腔、咽部分泌物及支气管肺泡灌洗液（bronchoal veolar lavage fluid,BALF）进行真菌培养、鉴定及随机扩增多态性DNA（random amplified polymorphic DNA,RAPD）基因分型分析。结果：在烟曲霉、黄曲霉和黑曲霉的RAPD带形图中,扩增条带数在1～11条之间,片断大小在150bp～2kb不等,不同菌株之间扩增条带的数量和扩增片断的大小均存在差异,即使带型完全一致的菌株,有些片断也存在亮度的差异。对分离自同一患者呼吸道不同部位的同种曲霉种内相似性比较的结果显示：虽然来自同一患者呼吸道不同部位的同种曲霉的形态学特征相同,但是基因型特征却存在差异。结论：通过呼吸作用进入肺组织内部的曲霉与患者鼻腔定植的同种曲霉存在差异,提示肺部曲霉的感染可能来源于患者周围的环境中,有效的控制环境中曲霉的污染程度是预防肺部曲霉感染的最有利的手段。     

The AgI/II Family Adhesin AspA Is Required for Respiratory Infection by Streptococcus pyogenes

Streptococcus pyogenes (GAS) is a human pathogen that causes pharyngitis and invasive diseases such as toxic shock syndrome and sepsis. The upper respiratory tract is the primary reservoir from which GAS can infect new hosts and cause disease. The factors involved in colonisation are incompletely known however. Previous evidence in oral streptococci has shown that the AgI/II family proteins are involved. We hypothesized that the AspA member of this family might be involved in GAS colonization. We describe a novel mouse model of GAS colonization of the nasopharynx and lower respiratory tract to elucidate these interactions. We used two clinical M serotypes expressing AspA, and their aspA gene deletant isogenic mutants in experiments using adherence assays to respiratory epithelium, macrophage phagocytosis and neutrophil killing assays and in vivo models of respiratory tract colonisation and infection. We demonstrated the requirement for AspA in colonization of the respiratory tract. AspA mutants were cleared from the respiratory tract and were deficient in adherence to epithelial cells, and susceptible to phagocytosis. Expression of AspA in the surrogate host Lactococcus lactis protected bacteria from phagocytosis. Our results suggest that AspA has an essential role in respiratory infection, and may function as a novel anti-phagocytic factor.     

Hydrating the respiratory tract: An alternative explanation why masks lower severity of COVID-19

The seasonality of respiratory diseases has been linked, among other factors, to low outdoor absolute humidity and low indoor relative humidity, which increase evaporation of water in the mucosal lining of the respiratory tract. We demonstrate that normal breathing results in an absorption-desorption cycle inside facemasks, in which supersaturated air is absorbed by the mask fibers during expiration, followed by evaporation during inspiration of dry environmental air. For double-layered cotton masks, which have considerable heat capacity, the temperature of inspired air rises above room temperature, and the effective increase in relative humidity can exceed 100%. We propose that the recently reported, disease-attenuating effect of generic facemasks is dominated by the strong humidity increase of inspired air. This elevated humidity promotes mucociliary clearance of pathogens from the lungs, both before and after an infection of the upper respiratory tract has occurred. Effective mucociliary clearance can delay and reduce infection of the lower respiratory tract, thus mitigating disease severity. This mode of action suggests that masks can benefit the wearer even after an infection in the upper respiratory tract has occurred, complementing the traditional function of masks to limit person-to-person disease transmission. This potential therapeutical use should be studied further.     

Nasal-associated lymphoid tissue is a mucosal inductive site for virus-specific humoral and cellular immune responses

Peyer's patches are known as mucosal inductive sites for humoral and cellular immune responses in the gastrointestinal tract. In contrast, functionally equivalent structures in the respiratory tract remain elusive. It has been suggested that nasal-associated lymphoid tissue (NALT) might serve as a mucosal inductive site in the upper respiratory tract. However, typical signs of mucosal inductive sites like development of germinal center reactions after Ag stimulation and isotype switching of naive B cells to IgA production have not been directly demonstrated. Moreover, it is not known whether CTL can be generated in NALT. To address these issues, NALT was structurally and functionally analyzed using a model of intranasal infection of C3H mice with reovirus. FACS and histological analyses revealed development of germinal centers in NALT in parallel with generation and expansion of IgA(+) and IgG2a(+) B cells after intranasal reovirus infection. Reovirus-specific IgA was produced in both the upper respiratory and the gastrointestinal tract, whereas production of reovirus-specific IgG2a was restricted to NALT, submandibular, and mesenteric lymph nodes. Moreover, virus-specific CTL were detected in NALT. Limiting dilution analysis showed a 5- to 6-fold higher precursor CTL frequency in NALT compared with a cervical lymph node. Together these data provide direct evidence that NALT is a mucosal inductive site for humoral and cellular immune responses in the upper respiratory tract.     

婴幼儿呼吸道微生物群演替研究进展

婴幼儿从出生开始正常呼吸道微生物就无时不刻地进行着演替,最终呼吸系统微生物形成动态平衡。婴幼儿时期是呼吸道微生物群演替的重要时期,也是免疫发育的关键时期,容易受外界因素的影响,这些影响因素包括分娩方式、喂养方式、抗生素、季节、疫苗接种等。研究表明婴幼儿时期的呼吸道微生物的组成和发育会影响微生物群的稳定性,从而影响呼吸道感染和过敏性疾病的发生。现对婴幼儿呼吸道微生物群演替及其影响因素,如分娩方式、喂养方式、抗生素使用、季节、疫苗接种等进行综述。     

武汉地区空气中花粉调查研究

空气中花粉粒的调查研究属气传孢粉应用学科,它正愈来愈受到临床医学界的重视。掌握本地区空气中花粉播散规律,对预防、诊断、治疗呼吸道过敏性疾病具有十分重要而现实的意义。本文着重报道了从1985年4月1日至1986年3月31日在武汉地区空气中孢粉散布的基本规律,并讨论分析了空气中花粉含量与气候的关系。根据花粉在空气中出现时期的长短及数量的多少,笔者把武汉地区空气中常遇到的一些植物花粉分为3种类型,这对考虑湖北武汉地区哪些植物花粉可能导致过敏性疾病是有意义的。     

Immunohistochemical demonstration of carbonic anhydrase isoenzyme VI (CA VI) expression in rat lower airways and lung.

Carbonic anhydrase isoenzyme VI (CA VI), which is transported in high concentrations in saliva and milk into the alimentary tract, is an important element of mucosal protection in the upper alimentary tract. Like alimentary tract mucosa, the respiratory tract mucosa is also exposed to heavy microbial, physical, and chemical stress. The protective and renewal-promoting factors present in the surface mucus of the respiratory tract are mainly produced by the seromucous tracheobronchial glands. Here we studied the secretion of CA VI by these glands in adult and developing rats using immunohistochemical techniques. The serous acinar and duct cells of the tracheobronchial glands stained for CA VI. The presence of the enzyme also in the duct content indicates its active secretion into the surface mucus. CA VI was also visible in the secretory cells and at the base of the ciliated cells of the tracheobronchial surface epithelium. Moreover, the Clara cells of the bronchiolar surface epithelium stained for CA VI. These findings are consistent with the hypothesis that CA VI has a mucosa-protective role not only in the gastrointestinal tract but also in the respiratory tract, where CA VI may act as a pivotal pH neutralizer and growth factor.     

Histological and ultrastructural specialization of the digestive tract of the intestinal air breather hoplosternum thoracatum (teleost)

The digestive tract of Hoplosternum thoracatum consists of an esophagus, gastric area, anterior digestive intestine with elaborate folds, digestive intestine with decreasing folds and thin, smooth-surfaced respiratory intestine. The upper tract has a mucoid columnar lining which is gently folded, whereas the gastric area has numerous pits opening into the tubular secretory glands. Striated muscle comprises the anterior muscularis but is replaced by inner circular and outer longitudinal smooth muscle layers in the gastric region. The digestive intestinal mucosa is elaborately folded, consisting of columnar cells with prominent brush borders. Mucosa, submucosa, circular and longitudinal muscularis and serosa layers are present throughout the tract. Goblet cells occur in both the digestive and respiratory intestine. Major changes that appear in the respiratory intestine are a drastic reduction in mucosa epithelial thickness and the penetration of an elaborate capillary bed into the epithelium. The other basic layers are not significantly reduced in thickness. The air-blood barrier consists of the thin epithelium, basement lamina and very thin capillary endothelium. Regional cellular composition and ultrastructural features are correlated with respective digestive and respiratory functions.     

Perspective on the host response to human metapneumovirus infection: what can we learn from respiratory syncytial virus infections?

Human metapneumovirus (HMPV) is a recently discovered pathogen first identified in respiratory specimens from young children suffering from clinical respiratory syndromes ranging from mild to severe lower respiratory tract illness. HMPV has worldwide prevalence, and is a leading cause of respiratory tract infection in the first years of life, with a spectrum of disease similar to respiratory syncytial virus (RSV). The disease burden associated with HMPV infection has not been fully elucidated; however, studies indicate that HMPV may cause upper or lower respiratory tract illness in patients between ages 2 months and 87 years, may co-circulate with RSV, and HMPV infection may be associated with asthma exacerbation. The mechanisms and effector pathways contributing to immunity or disease pathogenesis following infection are not fully understood; however, given the clinical significance of HMPV, there is a need for a fundamental understanding of the immune and pathophysiological processes that occur following infection to provide the foundation necessary for the development of effective vaccine or therapeutic intervention strategies. This review provides a current perspective on the processes associated with HMPV infection, immunity, and disease pathogenesis.     

金丝猴直肠脱的诊治一例

在四川省宝顶沟自然保护区内,一只离群的川金丝猴,根据临床症状和实验室检验,被确诊为因肠炎引起直肠脱出,并伴有上呼吸道感染。经过麻醉治疗、消除感染和驱虫后,该金丝猴呼吸道和消化道炎症消退,血蜱清除,肠道寄生虫驱除。再经过近两周的精心饲养,金丝猴被成功放回保护区。     

Linking influenza virus tissue tropism to population-level reproductive fitness

Influenza virus tissue tropism defines the host cells and tissues that support viral replication and contributes to determining which regions of the respiratory tract are infected in humans. The location of influenza virus infection along the respiratory tract is a key determinant of virus pathogenicity and transmissibility, which are at the basis of influenza burdens in the human population. As the pathogenicity and transmissibility of influenza virus ultimately determine its reproductive fitness at the population level, strong selective pressures will shape influenza virus tissue tropisms that maximize fitness. At present, the relationships between influenza virus tissue tropism within hosts and reproductive fitness at the population level are poorly understood. The selective pressures and constraints that shape tissue tropism and thereby influence the location of influenza virus infection along the respiratory tract are not well characterized. We use mathematical models that link within-host infection dynamics in a spatially-structured human respiratory tract to between-host transmission dynamics, with the aim of characterizing the possible selective pressures on influenza virus tissue tropism. The results indicate that spatial heterogeneities in virus clearance, virus pathogenicity or both, resulting from the unique structure of the respiratory tract, may drive optimal receptor binding affinity-that maximizes influenza virus reproductive fitness at the population level-towards sialic acids with α2,6 linkage to galactose. The expanding cell pool deeper down the respiratory tract, in association with lower clearance rates, may result in optimal infectivity rates-that likewise maximize influenza virus reproductive fitness at the population level-to exhibit a decreasing trend towards deeper regions of the respiratory tract. Lastly, pre-existing immunity may drive influenza virus tissue tropism towards upper regions of the respiratory tract. The proposed framework provides a new template for the cross-scale study of influenza virus evolutionary and epidemiological dynamics in humans.     

EPINEPHRINE HAND NEBULIZER IN ASTHMA—Technique of Use,Clinical Aspects

It behooves the physician seeking relief for asthmatic patients not to be casual about the epinephrine hand nebulizer and the manner in which it is used. Patients who claim to get no relief from the nebulizer should be asked to demonstrate their technique. If the nebulizer produces a hardly visible mist, it should be discarded. For many patients, the goal in spraying by hand must be the production of more or less continuous and voluminous aerosol, regardless of the phase of respiration, in order to effect relief. If the respiratory pattern has not deviated too far from normal in rate and depth, inhalations may be carried out in courses or cycles of about a half-minute duration and spaced a few minutes apart. If the respiratory pattern is abnormal during the act of spraying, it must be corrected.Inhaled epinephrine aerosols as constituted today appear to be somewhat irritating to the mouth, throat, and upper portion of the pulmonary tract of some persons, but it has not been convincingly demonstrated that serious and permanent damage to the lower respiratory tract of humans can occur from long-continued use of inhaled epinephrine.     

Epinephrine hand nebulizer in asthma; technique of use,clinical aspects

It behooves the physician seeking relief for asthmatic patients not to be casual about the epinephrine hand nebulizer and the manner in which it is used. Patients who claim to get no relief from the nebulizer should be asked to demonstrate their technique. If the nebulizer produces a hardly visible mist, it should be discarded. For many patients, the goal in spraying by hand must be the production of more or less continuous and voluminous aerosol, regardless of the phase of respiration, in order to effect relief. If the respiratory pattern has not deviated too far from normal in rate and depth, inhalations may be carried out in courses or cycles of about a half-minute duration and spaced a few minutes apart. If the respiratory pattern is abnormal during the act of spraying, it must be corrected. Inhaled epinephrine aerosols as constituted today appear to be somewhat irritating to the mouth, throat, and upper portion of the pulmonary tract of some persons, but it has not been convincingly demonstrated that serious and permanent damage to the lower respiratory tract of humans can occur from long-continued use of inhaled epinephrine.     

ELISA间接法检测呼吸道合胞病毒和副流感病毒血清IgM和IgA抗体

建立了检测呼吸道合胞病毒(RSV)和副流感病毒(PFV)血清特异性IgM和IgA抗体的间接ELISA方法。在方法统一的基础上比较了检测IgG、IgM和IgA抗体的结果,证明检测血清IgM和IgA可以作为RSV和PFV感染的早期诊断指标。检测了120份临床急性下呼吸道感染患儿的血清,RSV-IgM检出率为33.3％,RSV-IgA为36.7％;PFV-IgM为27.5％,PFV-IgA为31.6％。提出了对RSV和PFV感染以检测特异性IgA替代IgM或两者互补的设想。     

呼吸道感染与甲型链球菌的相关性研究

目的：探讨呼吸道感染与甲型链球菌的相关性。方法 对呼吸道感染患者和健康对照组口腔咽后壁取样进行分析。结果 健康对照组中甲型链球菌检出率（５０％）明显高于呼吸道感染病人组（２５．７５％）,且各疾病组甲型链球菌检出率均有显著性差异（Ｐ＜０．０５）,如肺内感染１６．６７％,支气管炎１７．２４％,具有显著性差异（Ｐ＞０．０５）,但肺炎患者甲型链球菌检出率５３．８５％与健康人相比无显著性差异（Ｐ＞０．０５）     

Bronchoscopy of cetaceans

Bronchoscopy is a standard diagnostic and therapeutic procedure in respiratory medicine and has been performed on many animal species. Cetaceans suffer considerable morbidity and mortality from lower respiratory tract infections, and it is very difficult to sample lower respiratory tract secretions for microbiology and other analyses. We report our experience on performing fiberoptic bronchoscopy in three bottle-nosed dolphins (Tursiops truncatus) and one false killer whale (Pseudorca crassidens), which should help other clinicians in performing bronchoscopy in cetaceans.     

Human Coronavirus HKU1 Infection of Primary Human Type II Alveolar Epithelial Cells: Cytopathic Effects and Innate Immune Response

Because they are the natural target for respiratory pathogens, primary human respiratory epithelial cells provide the ideal in vitro system for isolation and study of human respiratory viruses, which display a high degree of cell, tissue, and host specificity. Human coronavirus HKU1, first discovered in 2005, has a worldwide prevalence and is associated with both upper and lower respiratory tract disease in both children and adults. Research on HCoV-HKU1 has been difficult because of its inability to be cultured on continuous cell lines and only recently it was isolated from clinical specimens using primary human, ciliated airway epithelial cells. Here we demonstrate that HCoV-HKU1 can infect and be serially propagated in primary human alveolar type II cells at the air-liquid interface. We were not able to infect alveolar type I-like cells or alveolar macrophages. Type II alveolar cells infected with HCoV-HKU1 demonstrated formation of large syncytium. At 72 hours post inoculation, HCoV-HKU1 infection of type II cells induced increased levels of mRNAs encoding IL29,CXCL10, CCL5, and IL-6 with no significant increases in the levels of IFNβ. These studies demonstrate that type II cells are a target cell for HCoV-HKU1 infection in the lower respiratory tract, that type II alveolar cells are immune-competent in response to infection exhibiting a type III interferon and proinflammatory chemokine response, and that cell to cell spread may be a major factor for spread of infection. Furthermore, these studies demonstrate that human alveolar cells can be used to isolate and study novel human respiratory viruses that cause lower respiratory tract disease.