Nitric oxide deficiency contributes to impairment of airway relaxation in cystic fibrosis mice

The pulmonary disease of cystic fibrosis (CF) is characterized by persistent airway obstruction, which has been attributed to chronic endobronchial infection and inflammation. The levels of exhaled nitric oxide (NO) are reduced in CF patients, which could contribute to bronchial obstruction through dysregulated constriction of airway smooth muscle. Because airway epithelium from CF mice has been shown to have reduced expression of inducible NO synthase, we examined airway responsiveness and relaxation in isolated tracheas of CF mice. Airway relaxation as measured by percent relaxation of precontracted tracheal segments to electrical field stimulation (EFS) and substance P, a nonadrenergic, noncholinergic substance, was significantly impaired in CF mice. The airway relaxation in response to prostaglandin E2 was similar in CF and non-CF animals. Treatment with the NO synthase inhibitor NG-nitro-L-arginine methylester reduced tracheal relaxation induced by EFS in wild-type animals but had virtually no effect in the CF mice. Conversely, exogenous NO and L-arginine, a NO substrate, reversed the relaxation defect in CF airway. We conclude that the relative absence of NO compromises airways relaxation in CF, and may contribute to the bronchial obstruction seen in the disease.     

Transbronchial fine needle aspiration biopsy and rapid on‐site evaluation in the setting of superior vena cava syndrome

There is a paucity of prospective data on flexible bronchoscopy with rapid on‐site evaluation (ROSE) in the setting of superior vena cava (SVC) syndrome. The aims of this prospective study were to assess the diagnostic yield and safety of these investigations and specifically to evaluate the role of ROSE in limiting the need for tissue biopsies. Over a 5‐year period 48 patients (57.4 ± 9.7 years) with SVC syndrome secondary to intrathoracic tumors underwent flexible bronchoscopy with TBNA and ROSE. Endobronchial Forceps biopsy was reserved for visible endobronchial tumors with no on‐site confirmation of diagnostic material. ROSE confirmed diagnostic material in 41 cases (85.4%), and in only one of the remaining cases did the addition of a forceps biopsy increase the diagnostic yield (overall diagnostic yield of 87.5%). No serious complications were noted. The final diagnoses made included nonsmall lung cancer (n = 27), small cell lung cancer (n = 16), and metastatic carcinoma (n = 3). Two undiagnosed cases died of suspected advanced neoplasms (unknown primary tumors). We conclude that TBNA has a high diagnostic yield and is safe in the setting of SVC syndrome. With the addition of ROSE, tissue biopsy is required in the minority of cases. Diagn. Cytopathol. 2013;41:324–329. © 2011 Wiley Periodicals, Inc.     

MUC5AC and MUC5B Mucins Are Decreased in Cystic Fibrosis Airway Secretions

Cystic fibrosis (CF) is characterized by progressive airway obstruction. Although it has been postulated that this is due in part to mucus hypersecretion, there are no published data showing an increase in the gel-forming mucins MUC5AC or MUC5B in CF secretions. We used confocal microscopy to assess the amount of mucin-like glycoprotein and DNA in CF sputum and found more mucin in bronchitis sputum and a much greater amount of DNA in CF sputum. We then used antibodies to MUC5AC and MUC5B with Western gels and dot-blot to quantify mucin in sputum from 12 patients with CF and 11 subjects without lung disease. There was a 70% decrease in MUC5B and a 93% decrease in MUC5AC in CF sputum (P < 0.005 for both). We conclude that the vol/vol concentration of MUC5AC and MUC5B are decreased in the CF airways relative to normal mucus. This may be due to a relative increase in other components of sputum in the CF airway or to a primary defect in mucin secretion in CF.     

Does a primary host defense abnormality involving monocytes-macrophages underlie the pathogenesis of lung disease in cystic fibrosis?

The primary cause of morbidity and mortality in cystic fibrosis (CF) patients is chronic pulmonary disease. Pulmonary disease in CF is characterized in part by: (a) obstruction of the bronchi and bronchioles by inspissated secretions (mucus is hypersecreted and may also be abnormal), (b) recurrent or persistent bacterial infections, and (c) a chronic inflammatory state. We propose herein that much of the pathophysiology of lung disease in CF stems from a genetically inherited metabolic defect in monocyte-macrophages (M-MØ), and we review evidence which indicates that CF M-MØ are innately metabolically abnormal. Once activated by various stimuli, CF M-MØ become metabolically hyperactive and hypersecretory as evidenced by the production of excessive levels of a variety of mediators which could have definite roles in both the initiation of pulmonary obstruction and the accelerated development of a chronic inflammatory response in CF. Evidence is also reviewed which indicates that other CF M-MØ functions crucial to the afferent and efferent phases of the immune response to bacterial infections in the lung may be adversely affected. Mechanisms proposed to explain the abnormal production of mediators by CF M-MØ are discussed, and it is concluded that hyperproduction of mediators by CF M-MØ and their metabolic hyperactivity probably result from a defect in autoregulation. The nature of the metabolic defect in CF M-MØ indicates that CF should be classified as a “new” primary host defense abnormality or alternatively as a “new” primary immune deficiency disease.     

Sinus disease in cystic fibrosis

Conclusion Sinus abnormalities are prevalent in patients with CF. Symptoms such as nasal obstruction, loss of taste, and chronic headaches are common and often difficult to treat. When evaluating a patient with chronic sinusitis, the findings of noneosinophilic nasal polyps, unusual bacterial organisms, or specific radiographic features suggest the underlying diagnosis of CF. CF should be looked for even in the absence of pulmonary or gastrointestinal symptoms (34). Medical and surgical therapy reduces morbidity in CF patients, although much remains unknown about ideal therapeutic management.     

Hyperechogenic fetal bowel: counseling difficulties

The detection of echodense fetal bowel on ultrasound examination in the second trimester of pregnancy justifies invasive procedures such as amniocentesis to detect an underlying cause. We present a case in which initial tests identified only one mutation in the cystic fibrosis transmembrane regulator (CFTR)-gene of the fetus, the family history being negative for CF. Strongly reduced intestinal enzyme activities suggested intestinal obstruction and further increased the estimated risk for CF. After the 24th gestational week, a second mutation was found, confirming cystic fibrosis in this child. Problems in counseling in this particular case are discussed.     

Evaluation of direct immunofluorescence, enzyme immunoassay, centrifugation culture, and conventional culture for the detection of respiratory syncytial virus.

Four methods of detecting respiratory syncytial virus (RSV) from clinical specimens were evaluated. A total of 410 specimens consisting of nasopharyngeal washes, aspirates, and swabs were simultaneously tested for the presence of RSV by direct immunofluorescence assay (DFA), enzyme immunoassay (EIA) (Kallestad Pathfinder), shell vial centrifugation culture (SVC), and conventional culture. DFA identified 146 (83%) of the 175 positive cases, EIA detected 153 (87%), SVC detected 127 (73%), and conventional culture detected 70 (40%). Conventional culture isolated an additional 19 respiratory viruses other than RSV. DFA and EIA were able to detect nonviable virus not isolated by a culture method, and SVC isolated low-titer virus not detected by conventional culture. DFA and EIA gave similar results; however, the EIA system was less dependent on technical expertise. The use of SVC enhanced the conventional culture system with 63 RSV isolates not recovered from the tube culture. We recommend complementary use of both culture and nonculture methods in the detection of RSV.     

A Study of the Relationship of the Superior Vena Cava to the Bony Landmarks of the Sternum in the Supine Adult: Implications for Magnetic Guidance Systems

This study was a retrospective investigation of computed tomography (CT) images of an Australian adult hospital population. These images were used to evaluate the spatial relationships of the superior vena cava (SVC) to the midsagittal line, the sternal paracoronal plane, and commonly used landmarks of the sternum. Consistent relationships were found between the long axis of the SVC and both the midsagittal line and sternal paracoronal plane. When the sternal paracoronal plane was used as the plane of reference, the angle of Louis was found to approximate the SVC in 99.5% of cases, and the right 2nd intercostal space approximated the SVC in 94% of cases. Constraints on the use of landmarks are discussed with regard to magnetic guidance systems for catheter tracking.     

Atypical Cystic Fibrosis and CFTR-Related Diseases

Cystic fibrosis (CF), which is among the most common life-shortening recessive illnesses, is caused by mutations of the CF transmembrane conductance regulator (CFTR) and typically involves chronic infection and progressive obstruction of the respiratory tract as well as pancreatic exocrine insufficiency. Disease severity, to some extent, correlates with organ sensitivity to CFTR dysfunction and to the amount of functional protein, which is influenced by the type of mutation. Atypical CF represents approximately 2% of affected individuals, and includes cases presenting in adolescence or adulthood with pancreatic exocrine sufficiency, normal or borderline sweat chloride concentrations, or with a single predominant clinical feature. This review briefly describes diagnostic methods and phenotypic characteristics of classic and atypical CF, as well as CFTR-related diseases, conditions in which mutated CFTR may contribute to the pathogenesis but do not strictly fit established diagnostic criteria.     

Non-CFTR chloride channels likely contribute to secretion in the murine small intestine

While most cystic fibrosis (CF) transmembrane conductance regulator (CFTR)-knockout animals die due to intestinal obstruction before or at the time of weaning, a subpopulation of these animals are long living and exhibit a milder phenotype. The decreased severity of intestinal disease in these mildly affected CF mice is related to the expression of non-CFTR genetic modifiers. The identity of these genetic modifiers is not known, but we hypothesize that they may complement CFTR function as a chloride channel in this tissue. To assess the contribution of non-CFTR chloride channels to chloride secretion across the small intestine of CF mice with mild disease, we measured the basal transepithelial potential difference across this tissue as well as the secretory response to agonists of the cAMP and the calcium-mediated signaling pathways. Chloride secretion across the small intestine of mildly affected CF mice was not stimulated by forskolin or by carbachol. The absence of CFTR is thus not compensated by the activity of a distinct, cAMP- or calcium-activated chloride channel at the apical surface of the intestinal epithelium. On the other hand, a basal chloride secretion across the intestinal epithelium was present in these animals, and we hypothesize that this activity may be linked to improved survival of these animals.     

A 31-mutation assay for cystic fibrosis testing in the clinical molecular diagnostics laboratory

We devised a set of allele-specific probes to detect simultaneously 31 known cystic fibrosis mutations using PCR and the reverse dot blot detection format. The assay has been implemented in a clinical setting to the screening of over 750 individuals. Of these 102 Caucasians, 20 Hispanics and 1 Indian patient were affected with cystic fibrosis. The mutation detection rate in the 204 Caucasian and 40 Hispanic CF chromosomes was respectively, 88% and 85%. The availability of the probe sequences to CF screening laboratories should allow implementation of this assay in a clinical setting and comparison of its mutation typing rate among different centers. © 1995 Wiley-Liss, Inc.     

Innate inflammatory responses of pediatric cystic fibrosis airway epithelial cells: effects of nonviral and viral stimulation

There is controversy regarding whether cystic fibrosis (CF) airway epithelial cells (AECs) are intrinsically proinflammatory. The objective of the current study was to characterize the inflammatory profiles of AECs from children with CF compared with cells from healthy control subjects. We obtained AECs from healthy children (12) and children with CF (27). Biochemical and functional characteristics were assessed by stimulating cells with IFNγ, LPS, a cocktail referred to as cytomix, which consists of IFNγ, IL-1β, TNF-α, and LPS, or with human rhinovirus (HRV). Cytokine production was assessed using ELISA. Apoptotic responses to HRV infection were measured via production of single-stranded DNA. Our results indicated that CF and healthy cells exhibited similar morphology in monolayer culture. CF cells constitutively produced greater amounts of IL-6, IL-1β, and prostaglandin E(2), but similar levels of IL-8 and soluble intracellular adhesion molecule-1 compared with healthy cells, and this profile was maintained through repeated passage. Stimulation with LPS or cytomix elicited similar levels of IL-8 in CF and non-CF cells. In contrast, exposure to HRV1b resulted in a marked increase in IL-8 production from CF compared with non-CF cells. CF cells also exhibited reduced apoptosis and increased viral replication compared with non-CF cells after exposure to HRV1b. We conclude that CF and healthy AECs have similar basal and stimulated expression of IL-8 in response to proinflammatory stimuli, but elevated IL-8 release in response to HRV infection. The elevated IL-8, together with dampened apoptotic responses by CF cells to HRV, could contribute to augmented airway inflammation in the setting of recurrent viral infections early in life.     

Obstructive sleep apnoea secondary to superior vena cava thrombosis in a patient with activated protein C resistance

Summary Question of the Study Only five cases of superior vena cava (SVC) thrombosis as a cause of obstructive sleep apnoea (OSA) have as yet been reported. In this study, we aimed to describe an additional case and to review the literature on this subject.
Patients and Methods Based on the medical records, the case history of a 58-year-old man with OSA due to SVC thrombosis is presented. Furthermore, PubMed articles were browsed with the search items 'sleep apnoea' and 'superior vena cava thrombosis'.
Results The patient complained of the development of facial swelling and plethora during the previous 2  months. Furthermore, he reported the recent onset of snoring and excessive daytime sleepiness. Computed tomography of the thorax and mediastinal phlebography showed SVC thrombosis. Polysomnography demonstrated moderate to severe OSA. The cases of OSA secondary to SVC thrombosis already reported in the literature were mainly caused by mediastinal tumours compressing the SVC. In the present case, we found activated protein C (APC) resistance as the underlying disease. From the literature, it is known that therapies aimed at recanalization or relief of the SVC might lead to reversal of the OSA. In our patient, recanalization of the SVC was not possible and thus he had to be treated by continuous positive airway pressure therapy.
Conclusion In summary, we report on the rare occurrence of OSA due to SVC thrombosis. In the present case we found APC resistance as the underlying disease, which has not yet been reported in the literature.     

囊性纤维变性跨膜调节器基因突变对男性生殖的影响

1　ＣＦＴＲ基因突变及主要症状囊性纤维变性跨膜调节器 (ｃｙｓｔｉｃｆｉｂｒｏｓｉｓｔｒａｎｓ ｍｅｍｂｒａｎｅｃｏｎｄｕｃｔａｎｃｅｒｅｇｕｌａｔｏｒ,ＣＦＴＲ)基因于 1989年被发现 ,位于人的第 7染色体上。其编码的蛋白是一种膜蛋白 ,这种跨膜蛋白是一个Ｃｌ- 离子通道蛋白 ,受ｃＡＭＰ的调控。正常的ＣＦＴＲ由 14 80个氨基酸组成 ,含有两个结构区 ,每个结构区含有 6个跨膜片段[1] ,由称为Ｒ -结构域的多肽链连接 ,Ｒ -结构域上有大量的磷酸化位点 ,还有两个核苷酸结合位点(ｎｕｃｌｅｏｔｉｄｅｂｉｎｄｉｎｇｆｏｌｄｓ ,Ｎ…     

The role of inflammation in the pathophysiology of CF lung disease

Cystic fibrosis (CF) lung disease is characterized by a self-perpetuating cycle of airway obstruction, chronic bacterial infection, and vigorous inflammation that results in structural damage to the airway. CF patients have a predilection for infection with a limited spectrum of distinctive bacteria that initiate a vigorous inflammatory response which is more harmful than protective. The airway epithelial cell, which normally expresses the cystic fibrosis transmembrane conductance regulator (CFTR), directs the inflammatory response. Defects in CFTR are associated with increased production of proinflammatory mediators including IL-8, a potent neutrophil chemoattractant that stimulates the influx of massive numbers of neutrophils into the airways. These neutrophils are the primary effector cells responsible for the pathological manifestations of CF lung disease. Documented deficiencies in immuno regulatory molecules such as IL-10 likely contribute to the generation of the excessive and persistent inflammatory response. Since inflammation is a key contributor to the pathogenesis of CF lung disease, anti-inflammatory therapy must assume a larger role in CF until a cure is discovered. To date, attention has focused primarily on the therapeutic potential of systemic and inhaled corticosteroids and the non-steroidal anti-inflammatory drug (NSAID) ibuprofen. Development of new anti-inflammatory therapies that impact intracellular signaling pathways and cell-cell communication molecules likely will have the greatest impact on limiting the excessive production of the inflammatory mediators in the CF lung, thereby slowing the decline in lung function and improving survival.     

复方花刺参制剂对高脂模型大鼠血液流变学的改善

采用血液流变学检测方法 ,观察了复方花刺参制剂对高脂血症模型大鼠血粘度及其体外血栓的影响。结果发现此制剂具有明显的抗体外血栓形成和降低血粘度的作用 ,实验组与模型组大鼠在全血比粘度及体外血栓的长度、湿重和干重方面均存在着显著差异 (P <0 0 5 )。提示复方花刺参制剂能显著改善高脂模型大鼠的血液流变状态。     

Six-year molecular analysis of Burkholderia cepacia complex isolates among cystic fibrosis patients at a referral center for lung transplantation

Over a 6-year period, Burkholderia cepacia complex species were isolated from cystic fibrosis (CF) patients receiving care at The University of North Carolina Hospitals (clinic CF patients) and from those referred from other treatment centers. Fifty-six isolates collected from 30 referred patients and 26 clinic CF patients were characterized by pulsed-field gel electrophoresis (PFGE) and were assayed by PCR to detect the cable pilin gene, cblA. PFGE results indicated that six separate clusters (clusters A to F) were present among the 56 isolates and that three clusters (clusters A, B, and E) consisted only of isolates from referred patients infected with B. cepacia complex isolates prior to referral. However, one cluster (cluster C) consisted of isolates from four CF patients, and hospital records indicate that this cluster began with an isolate that came from a referred patient and that spread to three clinic CF patients. Cluster D consisted of two isolates from clinic CF patients, and hospitalization records are consistent with nosocomial, patient-to-patient spread. cblA was present in only 4 of the 56 isolates and included isolates in cluster E from the referred patients. Our results indicate a lack of spread of a previously characterized, transmissible clone from referred patients to our clinic CF population. Only two instances of nosocomial, patient-to-patient spread could be documented over the 6-year period. An additional spread of an isolate (cluster F) from a referred patient to a clinic patient could not be documented as nosocomial and may have been the result of spread in a nonhospitalized setting. The majority (36 of 56) of our B. cepacia complex-infected CF patients harbor isolates with unique genotypes, indicating that a diversity of sources account for infection. These data suggest that CF patients infected with B. cepacia complex and referred for lung transplantation evaluation were not a major source of B. cepacia complex strains that infected our resident CF clinic population.     

Bilateral thoracic ducts with coexistent persistent left superior vena cava

A case of bilateral thoracic ducts with coexistent persistent left superior vena cava (SVC) was identified in a 77-year-old Japanese female cadaver during dissection in a gross anatomy course. The persistent left SVC began at the lower surface of the left brachiocephalic vein, descended in front of the aortic arch, and drained into the right atrium through the coronary sinus. The right SVC was normal both in size and in position. The azygos vein, receiving the hemiazygos vein, opened into the right SVC. The accessory hemiazygos vein and the left superior intercostal vein united to form a common trunk, which drained into the left SVC. The left and right thoracic ducts began at the level of the 1st lumbar vertebra, ran upwards parallel and anterior to the vertebral column, and terminated at the venous angles of their corresponding sides. There was an anastomotic branch between them. The present case was considered to be very rare, since the persistent left SVC and bilateral thoracic ducts coexisted. The embryologic basis and clinical importance of this case are discussed.     

Improving clinical preimplantation genetic diagnosis for cystic fibrosis by duplex PCR using two polymorphic markers or one polymorphic marker in combination with the detection of the DeltaF508 mutation

Cystic fibrosis (CF) is an autosomal recessive disease characterized by obstruction and chronic infection of the respiratory tract and pancreatic insufficiency. The first preimplantation genetic diagnosis (PGD) for CF was carried out in 1992. At our centre the first cycle was performed in 1993. However, the number of known CF mutations is >1000, so developing mutation-specific PCR protocols for PGD is unfeasible. This is why a number of marker-based duplex PCRs were developed at the single cell level. A duplex PCR of a mutation and one or two microsatellites is not only a diagnostic tool, but it can also be used as a control for allele drop-out and contamination. During PGD, embryos obtained in vitro are analysed for the presence or absence of a particular genetic disease, after which only embryos shown to be free of this disease are returned to the mother. In total, 22 PGD cycles with duplex PCR (IVS8CA/IVS17BTA, DeltaF508/IVS8CA, DeltaF508/IVS17BTA and D7S486/D7S490) were carried out in 16 couples, which resulted in four ongoing pregnancies and one miscarriage.     

Stability-variability conflict as a reason for cancer: physical and social aspects.

We present an analysis of cancer based on the synthesis of physical and social models of life as the stability-variability conflict (SVC). We showed that the intensity of the SVC increases with an increase in the rate of evolution or progress of species and populations. The probability of oncogenesis depends on this intensity and on the gray zone in the host's homeostasis. An organism and its cells are two interdependent systems which in extreme situations become antagonistic and competitive. The hierarchy of the biosystems (cell, organism, population) does not resolve the SVC and does not decrease the gray zone of this conflict. Cancer develops mainly due to external effects and results in some internal, including genomic, alterations which result in cell death (apoptosis), or cause cells to begin a new stage in their lives, possibly leading to tumors. In this respect, cancer is a rescuer of the cells, although it brings about the death of the host. An increase in the production of tumor-associated proteins in cancer patients is an example of such changes. Cancer can also be analysed in terms of entropy and information. In the context of the SVC, cancer is irreversible deviations from the "golden mean" solution. The golden mean and cancer deviation are considered universal categories of physical and social reflections of the SVC as the quintessence of life.