The Effect of Filtration On the Cellular Components In Bronchoalveolar Lavage Fluid

The effect of filtration through two layers of cotton gauze on the cellular composition of bronchoalveolar lavage (BAL) fluid was studied in 25 patients with pulmonary disease. There was no significant difference between median total cell count in unfiltered and filtered BAL (P= 0.73) or in the distribution of neutrophils or eosinophils. the percentage of bronchial epithelial cells was significantly higher in unfiltered than in filtered BAL (P= 0.02). Furthermore, differential cell counts showed a significantly lower percentage of alveolar macrophages (P= 0.04), and a significantly higher percentage of lymphocytes (P=0.04) in unfiltered compared with filtered BAL. Thus, gauze filtration results in a loss of bronchial epithelial cells and lymphocytes, and is not recommended in the routine analysis of cellular components in BAL fluid. L'effet de la filtration à travers deux couches de gaze de coton sur la composition cellulaire du liquide de lavage broncho-alvéolaire (LBA) a étéétudié chez 25 patients atteints d'une pathologie pulmonaire. Aucune différence significative entre les échantillons filtrés et non filtrés n'a été trouvée en ce qui concerne la numération cellulaire totale moyenne (P=0.73) et la distribution des polynucléaires neutrophiles et eosinophiles. Le pourcentage de cellules épithéliales bronchiques est significativement plus élevé dans les échantillons non filtrés que dans ceux qui ont été filtrés (P= 0.02). Par ailleurs, le comptage differentiel montre un pourcentage plus faible de macrophages alveolaires (P=0.04) et un pourcentage plus élevé de lymphocytes (P= 0.04) dans les LBA non filtrés. La filtration à travers la gaze a pour résultat une perte des cellules épithéliales bronchiques et des lymphocytes et n'est donc pas à recommander pour l'analyse de routine des composants cellulaires du liquide de lavage broncho-alvéolaire. Die Auswirkung einer Filtration von BAL-Material durch doppelte Baumwollgaze wurde am Material 25 Patienten untersucht. Es bestand kein signifikanter Unterschied der Zellzahl von gefiltertem und ungefiltertem Material (P= 0.73) oder im Abteil der Neutrophilen und Eosinophilen. Das Abteil des Bronchial epithelien war jedoch in ungefiltertem Material signifikant höher (P= 0.02). Differentialauszählungen zeigten einen signifikant geringeren Anteil an Alveolarmakrophagen (P= 0.04) und einen signifikant erhöhten an Lymphozyten (P= 0.04) in unfil-triertem Material. Die Filtration führt also zu einem Verlust an Bronchialepithelien und Lymphozyten und ist deshalb für Routine-untersuchungen von BAL nicht zu empfehlen.     

Bronchoalveolar Lavage In the Diagnosis of Bronchiolo-Alveolar Carcinoma. A Case Report

Bronchiolo-alveolar carcinoma is usually localized to the terminal bronchioles and alveoli, and may present on chest X-ray as interstitial pulmonary disease with diffuse reticulonodular infiltrates. The cytological diagnosis is often difficult to obtain. This case demonstrates that bronchoalveolar lavage can be useful in the diagnostic evaluation of this type of malignancy, in conjunction with transbronchial lung biopsy.     

The Diagnosis of Pneumocystis Carinii Pneumonia By Cytologic Evaluation of Papanicolaou and Leishman-Stained Bronchoalveolar Specimens In Patients With the Acquired Immunodeficiency Syndrome

The presence of foamy alveolar casts or flocculent material in Papanicolaou and Leishman-stained smears of bronchoalveolar lavage (BAL) fluid is said to be indicative of infection with Pneumocystis carinii. We have investigated the sensitivity and specificity of this method of diagnosing pneumocystis pneumonia in patients with the acquired immunodeficiency syndrome (AIDS). Patients (n= 114) with diffuse lung infiltrates were submitted to fibreoptic broncoscopy and BAL. Seventy of them were patients with AIDS. the other 44 individuals were not infected by the human immunodeficiency virus (HIV). Pneumocystis carinii organisms were identified on Grocott's methenamine silver (GMS)-stained BAL smears in 30 patients with AIDS. Flocculent material was present in the Papanicolaou and Leishman-stained smears from all of these cases. Conversely, P. carinii were not seen on GMS-stained smears in the remaining 84 individuals with or without AIDS. No flocculent material was observed in Papanicolaou or Leishman-stained smears in these 84 patients. We concluded that the presence of flocculent material in Papanicolaou or Leishman-stained smears of BAL fluid is indicative of P. carinii pneumonia in patients with AIDS. La présence de cylindres alvéolaires spumeux ou de matériel floculé dans les étalements de liquide de lavage bronchoalvéolaire (LBA) colorés selon Papanicolaou ou Leishman est considérée comme symptomatique d'une infection par Pneumocystis carinii. Nous avons étudié la sensibilité et la spécificité de cette méthode de diagnostic de l'infection par Pneumocystis carinii chez des patients atteints de syndrome de déficience immunitaire acquise (SIDA). Cent quatorze malades avec des infiltrats pulmonaires diffus ont subi une fibroscopie bronchique et un lavage broncho-alvéolaire. Soixante dix d'entre eux edtaient atteints de SIDA, 44 n'étaient pas infectés par le Virus de l'Immunodéficience Humaine (VIH). Le Pneumocystis carinii a été identifiié par la coloration de Grocott chez 30 patients atteints de SIDA. Chez ces patients, la présence d'un matériel floculé est constante sur les étalements colorés au Papanicolaou et au Leishman. A l'inverse, Pneumocystis carinii n'a pas été retrouvé chez les 84 autres malades, atteints ou non du SIDA et les étalements de LBA ne contenaient pas de matériel floculé. En conclusion, la présence de matériel floculé dans les étalements de LBA colorés selon Papanicolaou ou Leishmanest associée à une pneumpathie àPneumocystis carinii chez les patients atteints de SIDA. Sensitivität und Spezifität des Nachweises schaumiger oder flockiger Alveolarausgüsse bei Pneumocystis carinii wurden in 114 Fällen diffuser Lungeninfiltrate untersucht. 70 Patienten waren an AIDS erkrankt, 44 weitere waren HIV-negative. In 30 der AIDS-Fälle wurde P. carinii mit der Grocott'schen Färbung nachgewiesen. Die typischen Eiweißniederschläge waren in all diesen Fällen nachweisbar. Umgekehrt ergab die Grocottfärbung in 84 Fällen mit oder ohne AIDS ein negatives Ergebnis. In all diesen Fällen war kein Eiweißniederschlag nachweisbar. Daraus ergibt sich, daß die Eiweißniederschläge in Präparaten, die nach Papanicolaou oder Leishman gefärbt wurden, kennziechned sind für die P. carinii Pneumonie.     

Identification of Phagocytosed Pneumocystis Carinii In Human Pulmonary Alveolar Macrophages

Examination of Papanicolaou-stained bronchoalveolar lavage samples from cases with Pneumocystis carinii pneumonitis under ultra-violet light reveals alveolar macrophages packed with fluorescent inclusions. Immunoenzymatic staining of the alveolar macrophages with a monoclonal antibody specific for P. carinii (3F6) showed that these inclusions contain intact pneumocysts or their degradation products. Fluorescence microscopy of Papanicolaou-stained smears is advocated as a sensitive and specific method of diagnosing P. carinii infection.     

肺泡灌洗技术对重症支原体肺炎患儿血清炎症因子及肺功能的影响

目的:探讨支气管肺泡灌洗液(Bronchoalveolar Lavage Fluid,BALF)技术应用于儿科重症支原体肺炎(Severe Mycoplasma Pneumoniae Pneumonia,SMPP)对患儿的血清炎症因子表达影响及肺功能的变化,旨在为临床上SMPP的诊治提供科学参考。方法:回顾性选取我院收治的SMPP患儿83例,按照治疗方式不同,分为两组,对照组41例,静滴阿奇霉素,研究组42例,采用静滴阿奇霉素3-5天行BALF灌洗生理盐水+布地奈德治疗。于治疗7 d后对比两组治疗前后及组间肺功能和炎症指标的变化。结果:研究组IL-2、IL-6、IL-10表达水平均较对照组显著降低,差异有统计学差异(P0.05);研究组mPAP、PAWP、PVR均比对照组下降明显,差异有显著性意义(P0.05)。结论:BALF技术可明显改善患儿炎症因子表达,对促进患儿肺功能恢复正常有良好作用,适用于儿童SMPP的治疗。     

支气管镜下肺泡灌洗对难治性肺炎患者炎症介质和肺功能的影响

目的:探讨支气管镜下肺泡灌洗对难治性肺炎患者炎症介质和肺功能的影响。方法:选择2013年1月~2015年12月在我院进行诊治的难治性肺炎患者100例,随机分为两组。对照组采取常规的临床治疗,观察组在常规治疗基础上采用支气管镜联合支气管肺泡灌洗治疗。观察两组的治疗效果以及对患者炎症介质和肺功能的影响。结果:观察组的有效率为90.00%(45/50),明显高于对照组的76.00%(38/50)(P0.05)。治疗后,两组肺泡灌洗液中的内毒素、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)和脂质过氧化物(LPO)水平均较治疗前明显降低(P0.05),且观察组降低的更为明显(P0.05);两组的肺功能均较治疗前明显改善(P0.05),且观察组改善的更为明显(P0.05)。结论:支气管镜下肺泡灌洗对难治性肺炎患者具有较好的治疗效果,可以有效降低炎症介质水平,并改善患者肺功能。     

Cut‐off values and significance of Oil Red O‐positive cells in bronchoalveolar lavage fluid

C. Basset‐Léobon, L. Lacoste‐Collin, J. Aziza, J.C. Bes, S. Jozan and M. Courtade‐Saïdi
Cut‐off values and significance of Oil Red O‐positive cells in bronchoalveolar lavage fluid Objective: To evaluate the percentage and predictive value of Oil Red O‐positive macrophages (ORO‐PM) to identify lipid‐laden macrophages in bronchoalveolar lavage fluids (BALF) from patients with different pathologies. Methods: The percentage and absolute numbers of ORO‐PM were evaluated in 305 BALF. The patients were separated into ten groups: corticosteroid treatment (n = 18), amiodarone treatment (n = 8), interstitial fibrosis (n = 11), human immunodeficiency virus (HIV)‐positive (n = 25), infectious pneumonia (n = 43), severe haematological disorder (n = 25), interstitial syndrome (n = 109), suspicion of cancer (n = 17), transplant recipients (n = 50) and controls (n = 43). The total and differential cell counts in BALF were recorded. The presence of specific pathogens was also noted. Parametric and non‐parametric tests were used to compare the values between groups. Receiver–operating characteristics (ROC) curves were established in order to determine a cut‐off value. Results: The percentages of ORO‐PM were (mean ± standard deviation) 21.67 ± 29.12 in the corticosteroid group, 10.00 ± 12.49 in the amiodarone group, 19.45 ± 20.72 in the interstitial fibrosis group, 47.80 ± 30.46 in the HIV group, 19.72 ± 26.26 in the infectious pneumonia group, 27.42 ± 30.04 in the severe haematological disorder group, 25.18 ± 30.63 in the interstitial syndrome group, 17.64 ± 27.76 in the suspicion of cancer group, 22.50 ± 27.27 in the transplanted recipients group and 2.63 ± 3.48 in the control group. Significantly higher values were found in all groups when compared with the control group (P < 0.001). Only the HIV group showed higher numbers of ORO‐PM when compared with the interstitial syndrome group (P < 0.01). According to ROC curves, > 6% ORO‐PM was suggested as the positive cut‐off value. Conclusion: Significantly increased numbers of ORO‐PM were associated with various lung pathologies. However, the higher numbers observed in HIV patients require further investigations.     

Quantitative Differences In Phagocytosis and Degradation of Pneumocystis By Alveolar Macrophages In Aids and Non-Hiv Patients In Vivo

Broncholaveolar lavage (BAL) specimens (n= 213) from AIDS and non-HIV immunosuppressed patients were investigated for the presence of Pneumocystis carinii infection by fluorescence microscopy of Papanicolaou-stained slides. Alveolar casts, extracellular pneumocysts and phagocytosed cysts and their degradation products in pulmonary alveolar macrophages were identified. the number of phagocytosed pneumocysts within human pulmonary alveolar macrophages was recorded and correlated with the number of extracellular cysts and alveolar casts, in both groups of patients. Both phagocytic and degradation capacity were depressed in AIDS patients. This observation may explain the large number of extracellular organisms found in BAL specimens of AIDS patients compared with non-HIV-positive immunocompromised individuals.     

Proteomic analysis of human bronchoalveolar lavage fluid: expression profiling of surfactant-associated protein A isomers derived from human pulmonary alveolar proteinosis using immunoaffinity detection

Human bronchoalveolar lavage fluid (BALF) proteins from pulmonary alveolar proteinosis (PAP) obtained by washing the epithelial lining of the lung with phosphate-buffered saline, were separated using high resolution two-dimensional gel electrophoresis (2-DE) under denaturing and reducing conditions. By Western blotting, the proteins were transferred from polyacrylamide gel onto a chemical resilient membrane. The surfactant-associated protein A (SP-A) isomers were then identified with enhanced chemiluminescence detection (ECL) using antibody-antigen reaction. Some of the gels were treated with silver staining after 2-DE. The molecular masses of SP-A isomers in BALF from PAP ranged from 20.5 to 26, 26 to 32, and 32 to 42 kDa, respectively; and isoelectric points (pI) were in pH range of 4.5-5.4 under denaturing and reducing conditions. In the mass range of 20.5-26 kDa and pI of 4.5-5.4, there were five isomers, and in mass range of 26-32 kDa and pI of 4.5 to 5.4, there were at least eight isomers on the ECL detection film. However, in the mass range of 32-42 kDa and pI of 4.5-5.4, there were three isomers separated one from another but there was also a cluster of overlapping spots on the ECL detection film. Thus, this communication describes a characteristic 2-DE pattern of SP-A isomers in BALF from PAP as follows. (1) The five isomers of mass 20.5-26 kDa and pI of 4.5-5.4; (2) the eight isomers of mass 26-32 kDa and pI of 4.5-5.4; and (3) the three isomers of mass 32-42 kDa and pI of 4.5-5.4.     

The Bovine Lung in Biomedical Research: Visually Guided Bronchoscopy,Intrabronchial Inoculation and In Vivo Sampling Techniques

There is an ongoing search for alternative animal models in research of respiratory medicine. Depending on the goal of the research, large animals as models of pulmonary disease often resemble the situation of the human lung much better than mice do. Working with large animals also offers the opportunity to sample the same animal repeatedly over a certain course of time, which allows long-term studies without sacrificing the animals.The aim was to establish in vivo sampling methods for the use in a bovine model of a respiratory Chlamydia psittaci infection. Sampling should be performed at various time points in each animal during the study, and the samples should be suitable to study the host response, as well as the pathogen under experimental conditions.Bronchoscopy is a valuable diagnostic tool in human and veterinary medicine. It is a safe and minimally invasive procedure. This article describes the intrabronchial inoculation of calves as well as sampling methods for the lower respiratory tract. Videoendoscopic, intrabronchial inoculation leads to very consistent clinical and pathological findings in all inoculated animals and is, therefore, well-suited for use in models of infectious lung disease. The sampling methods described are bronchoalveolar lavage, bronchial brushing and transbronchial lung biopsy. All of these are valuable diagnostic tools in human medicine and could be adapted for experimental purposes to calves aged 6-8 weeks. The samples obtained were suitable for both pathogen detection and characterization of the severity of lung inflammation in the host.     

Assessment of the Accuracy of Cytology In Women Referred For Colposcopy and Biopsy: the Results of a 1 Year Audit

The results of weekly colposcopy review meetings have been audited for 1 year and cases where there was a discrepancy between the referral cervical smear and the initial colposcopy biopsy have been analysed. New referrals (n = 476) for colposcopy were studied. In the final outcome 80% of 326 women referred for moderate or severe dyskaryosis were found to have cervical intraepithelial neoplasia (CIN) grade II or III or invasive carcinoma. Three women found to have invasive carcinoma had been referred for severely dyskaryotic smears. Twenty women were referred for smears with cell changes suggesting glandular neoplasia: five were found to have adenocarcinoma in situ, whereas eight had CIN and seven had negative biopsies. The results justify the referral policy and demonstrate the need for further investigation when initial colposcopic biopsies are negative.     

高压氧联合支气管肺泡灌洗对重型颅脑损伤肺部感染患者临床疗效及血清sTREM-1、HMGB1、CRP/Alb水平的影响

摘要 目的：探讨高压氧（HBO）联合支气管肺泡灌洗对重型颅脑损伤肺部感染（PI）患者临床疗效及血清可溶性髓系细胞触发受体-1（sTREM-1）、高迁移率族蛋白B1（HMGB1）、C反应蛋白/白蛋白（CRP/Alb）水平的影响。方法：选取2019年9月-2022年9月安徽中医药大学附属六安医院收治的72例重型颅脑损伤PI患者为研究对象,按随机数字表法分为观察组和对照组,每组各36例。对照组采用支气管肺泡灌洗治疗,观察组采用HBO联合支气管肺泡灌洗治疗。收集两组临床资料,对比两组临床疗效、治疗前后临床体征[体温、动脉血氧分压（PaO₂）,血白细胞计数（WBC）]、临床肺部感染评分（CPIS）、全身炎症反应综合征修正（ASS）评分、格拉斯哥昏迷评分（GCS）、血清指标（sTREM-1、HMGB1、CRP/Alb）。结果：观察组治疗总有效率94.44%高于对照组的77.78%（P＜0.05）;治疗后观察组体温、WBC、CPIS评分、ASS评分低于对照组,PaO₂、GCS评分高于对照组（P＜0.05）;治疗后观察组血清sTREM-1、HMGB1、CRP/Alb水平低于对照组（P＜0.05）。结论：HBO联合支气管肺泡灌洗治疗重型颅脑损伤PI能减轻炎症反应,降低PI程度,减轻临床体征,提高临床疗效。     

Role of sphingosine 1-phosphate and lysophosphatidic acid in fibrosis

This review highlights an emerging role for sphingosine 1-phosphate (S1P) and lysophosphatidic acid (LPA) in many different types of fibrosis. Indeed, both LPA and S1P are involved in the multi-process pathogenesis of fibrosis, being implicated in promoting the well-established process of differentiation of fibroblasts to myofibroblasts and the more controversial epithelial–mesenchymal transition and homing of fibrocytes to fibrotic lesions. Therefore, targeting the production of these bioactive lysolipids or blocking their sites/mechanisms of action has therapeutic potential. Indeed, LPA receptor 1 (LPA₁) selective antagonists are currently being developed for the treatment of fibrosis of the lung as well as a neutralising anti-S1P antibody that is currently in Phase 1 clinical trials for treatment of age related macular degeneration. Thus, LPA- and S1P-directed therapeutics may not be too far from the clinic. This article is part of a Special Issue entitled Advances in Lysophospholipid Research.