Reactivity of anti-Nipah virus monoclonal antibodies to formalin-fixed, paraffin-embedded lung tissues from experimental Nipah and Hendra virus infections

The immunohistochemical reactivity of seven clones of mouse monoclonal antibodies raised to Nipah virus antigens were investigated using formalin-fixed, paraffin embedded porcine and equine lung tissues from experimental Nipah and Hendra virus infection, respectively. Either microwave irradiation or enzymatic digestion effectively unmasked the viral antigens in formalin-fixed, paraffin-embedded tissue sections. Four clones showed positive reaction to both Nipah virus-infected porcine lung tissue and Hendra virus-infected equine lung tissue. Two clones (11F6 and 13A5) reacted with Nipah virus-infected porcine lung tissue, but not with Hendra virus-infected equine lung tissue. These Nipah virus-specific monoclonal antibodies may therefore be useful for immunohistological diagnosis of Nipah virus infection and for further research on Nipah virus pathogenesis.     

Evaluation of a thoracoscopic technique using ligating loops to obtain large lung biopsies in standing healthy and heaves-affected horses

Objective— To evaluate use of pre‐tied ligating loop to perform thoracoscopic, large lung biopsy in normal and heaves‐affected horses. Study Design— Prospective clinical study. Animals— Normal (n=5) and heaves‐affected (n=6) horses. Methods— Lung biopsies, 1 from each hemithorax, were collected thoracoscopically using a pre‐tied ligating loop. Horses were either normal (C) or heaves‐affected with the latter being in remission (Ha) for the initial biopsy and in exacerbation (Hs) for the 2nd biopsy. Clinical variables, PaO₂, and PaCO₂ were used to determine the effect of surgical biopsy. Postoperative pneumothorax was monitored by serial thoracic radiographic examinations. Results— Thoracoscopic lung biopsy (n=29, 22 procedures) was well tolerated by all horses. Complication rate was 31%, including 8 ligature slippage and 1 pulmonary hemorrhage. Intranasal oxygen was administered intraoperatively to 6 horses (2 C, 1 Ha, 3 Hs) with severe hypoxemia or labored breathing. There was a significant decrease in PaO₂ during surgery in horses not supplemented with oxygen. Postoperative pneumothorax (21/22 procedures) detected radiographically resolved within 3 weeks. Conclusion— Thoracoscopic lung biopsy using pre‐tied ligating loops was minimally invasive, relatively inexpensive, and fairly efficient. Heaves‐affected horses tolerated the surgery well, even when in exacerbation; however, the technique was associated with non life‐threatening complications in 31% of the biopsies, most of which required correction with additional ligating loops or more sophisticated instrumentation. Clinical Relevance— Using laparoscopic pre‐tied ligating loop for thoracoscopically‐assisted lung biopsy can be considered in horses with normal and impaired lung function but alternative instrumentation and access to intranasal oxygen must be available to the surgeon in case of complications.     

Persistence of Neospora caninum-specific immunoglobulin G antibodies in bovine blood and lung tissue stored at room temperature.

Blood and tissue samples obtained at necropsy from carcasses showing varying degrees of postmortem decomposition are sometimes used as sources of antibodies for serological tests, primarily in studies involving wild animals or in examination of aborted fetuses. To test the persistence of antibodies in blood and lung tissue kept at room temperature (approximately 20degreesC) for up to 48 and 118 days, respectively, serum and lung fluid from Neospora caninum-infected and uninfected cattle were tested using an indirect N. caninum immunostimulating complex enzyme-linked immunosorbent assay (ELISA). Testing of selected samples was also performed using a commercial Neospora competitive ELISA. Despite severe changes in the gross appearance of the samples, the blood collected through venipuncture from live animals remained essentially sterile, whereas the pieces of lung tissue showed a variety of bacterial and fungal growth. Nevertheless, there was no obvious reduction in antibody levels in either the blood or the tissue samples during the 2 observation periods. This study indicates that specific immunoglobulin G antibodies may be remarkably stable in sterile blood and decomposed lung tissue.     

Protective Effects of Rosmarinic Acid on Oxidative Lung Damage in Asthmatic Mice

In order to evaluate the protective effect of rosmarinic acid on oxidative lung damage in asthmatic mice, in this study, the asthma model was established by sensitizing and challenging female BALB/c mice with ovalbumin (OVA), and some mice were challenged by OVA and H₂O₂ as a positive control of oxidative lung damage. The BALF, lung tissues of mice were harvested 24 h after the final OVA challenge and the number of total cells and eosinophils were measured, ROS,SOD and GSH-Px levels were detected, and pathological changes of lung tissue were observed using HE staining. The results showed that rosmarinic acid significantly reduced the number of total cells and eosinophils in BALF, significantly inhibited the production of ROS in lung tissue and BALF, increased the levels of SOD and GSH-Px, and ameliorated pathological changes of lung tissue. The results suggested that rosmarinic acid could protect oxidative lung damage in asthmatic mice.     

迷迭香酸对哮喘小鼠氧化性肺损伤的保护作用

为评价迷迭香酸对哮喘小鼠模型氧化性肺损伤的保护作用,本研究用卵清蛋白（OVA）致敏、激发雌性BALB/c小鼠建立哮喘模型,并用OVA和H₂O₂联合激发小鼠作为氧化肺损伤阳性对照模型。在最后一次滴鼻激发24 h后,取支气管肺泡灌洗液（BALF）进行细胞计数并测定活性氧（ROS）、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）水平,取左侧肺脏固定做HE染色。结果显示,迷迭香酸可明显减少BALF中细胞总数和嗜酸性粒细胞数目,显著抑制肺组织和BALF中ROS的产生,升高SOD和GSH-Px水平,改善肺组织病理变化。本试验结果表明,迷迭香酸对氧化肺损伤起到明显的保护作用。     

Detection of respiratory pathogens in porcine lung tissue and lavage fluid

The objective of this study was to compare the detection rate of bacterial agents in bronchoalveolar lavage fluid (BALF), taken without visual control, to that in affected lung tissue obtained from the same pig at necropsy. BALF and affected lung tissue were examined for Mycoplasma hyopneumoniae using PCR, and standard cultural methods were used for Actinobacillus pleuropneumoniae, Bordetella bronchiseptica, Haemophilus parasuis, Pasteurella multocida and Streptococcus suis. All pigs with a history of respiratory symptoms were submitted as live animals for routine diagnostic examination. In each animal the site of lavage, marked by injecting methylene blue, differed from the site of pneumonic lesions. M. hyopneumoniae was detected more frequently in lung tissue than in BALF in cases with moderate or severe lung lesions. The detection rates of M. hyopneumoniae were higher in the BALF of pigs with mild lesions. Cultural examination of BALF was at least as satisfactory as affected lung tissue for detecting B. bronchiseptica, H. parasuis and P. multocida.     

Accidental entrapment of an endo-bronchial blocker tip by a surgical stapler during selective ventilation for lung lobectomy in a dog

OBJECTIVE: To describe the use of an endobronchial blocker (EBB) and to perform selective ventilation during pulmonary lobe resection via thoracotomy in a dog and report its accidental stapling in the resection site. STUDY DESIGN: Clinical case report. ANIMAL: One female dog with a suspected abscess or neoplasia of the right caudal pulmonary lobe. METHODS: One-lung ventilation was performed using a wire-guided EBB to seal the contaminated parenchyma and facilitate surgical access. The affected lung parenchyma was resected and the resection site was closed with staples. RESULTS: Lobar resection was performed successfully, but the loop of the EBB guide wire was inadvertently entrapped in the staple line of the lobectomy. Staples were removed to release the wire loop, and the resulting air leak caused loss of ventilation control until the parenchyma was re-sealed. CONCLUSIONS: We recommend removing the wire guide associate with the EBB after successful lung separation to avoid accidents that could have life-threatening consequences if not recognized. CLINICAL RELEVANCE: One-lung ventilation is useful to isolate healthy parenchyma from diseased parenchyma during lobectomy. Anesthesiologists and surgeons need to be aware of the potential complications associated with use of EBB.     

Computed tomography (CT) of the lungs of the dog using a helical CT scanner, intravenous iodine contrast medium and different CT windows

The aim of this study was to determine the accuracy of helical computed tomography (CT) for visualizing pulmonary parenchyma and associated formations in normal dogs. CT scan was performed by using intravenous contrast medium and by applying different types of CT windows: soft tissue and lung windows, and high-resolution computed tomography of the lung. This technique allowed, especially with lung window types, a good view of the parenchyma, bronchial tree, vascular structures and pleural cavity. The selected images, with high anatomical quality and tissue contrast, may be a reference for future clinical studies of this organ. Thus, helical CT is a promising non-invasive method of diagnosing a wide variety of pulmonary diseases in dogs.     

高原牦牛胎盘粉对小鼠部分组织中MDA含量的影响

将100只小白鼠随机分为四个组（高海拔组、中海拔组、低海拔组和对照组）,连续28d分别灌服不同海拔的牦牛胎盘粉和生理盐水,28d后采取肝、肺、心肌、骨骼肌、脾脏和肾脏,测定各组织内MDA的含量。结果表明牦牛胎盘粉能够降低小鼠肺脏、肝脏、肾脏、骨骼肌中MDA的含量。     

Detection of Burkholderia cepacia DNA from artificially infected EDTA-blood and lung tissue comparing different DNA isolation methods

Bacterial DNA (Burkholderia cepacia) was prepared from artificially infected equine ethylenediaminetetraacetic acid (EDTA)-blood and lung tissue by using four standard methods (lysis buffer containing proteinase K, phenol/chloroform/isoamylalcohol-extraction, microwave-treatment, heat treatment) and six commercially available kits (Puregene, High Pure PCR Template Preparation Kit, InstaGene, QiaAmp Tissue Kit, DNAzol and Elu-Quik). After a subsequent polymerase chain reaction (PCR), their efficacy and sensitivity were compared. Concerning the detection limits, the simple lysis with a proteinase K-containing buffer led to the best results for EDTA-blood as well as for artificially infected lung tissue.     

Detection of toxoplasmosis in experimentally infected goats by PCR

PCR was used to diagnose toxoplasmosis in two pairs of Barbari goats infected by oral administration of doses of either 10(4) or 10(5) oocysts of Toxoplasma gondii. Blood and lymph node aspirates were collected from the infected goats and control goat at intervals, and tissues were also collected from a fetus that was aborted and a doe that died during the trial. Both processed and unprocessed samples were used for the PCR, using primers directed to the multicopy B1 gene. None of the blood samples was positive, but a specific signal was obtained from the lymph node aspirates after partial DNA extraction. Direct PCR of the lung, muscle and mesenteric lymph node of the doe and lung tissue of the aborted fetus yielded the target fragment. The simplified PCR protocols, including partial DNA extraction and direct assay of lung tissue, were effective for the diagnosis of toxoplasmosis.     

青海藏獒尘肺的矿物分析

采用光镜、电子探针微分析仪及原子发射光谱(ICP),对购自青海省果洛藏族自治州达日县5例确诊为尘肺的藏獒肺脏进行了研究。结果表明,5例藏獒肺脏均有程度不等的尘肺病变,肺脏尘肺病变可表现为局灶型或弥漫型。局灶型,尘灶与尘细胞结节呈星散状分布,多位于细支气管周围、肺泡隔和肺胸膜结缔组织中;弥漫型则尘灶与尘细胞弥漫性分布于大范围肺组织中;并可见巨噬细胞性肺炎。元素分析表明,肺和淋巴结中的粉尘多为铝硅酸盐类化合物。     

Immune response to pulmonary injection of Pasteurella haemolytica-impregnated agar beads followed by transthoracic challenge exposure in goats

A method of inducing Pasteurella haemolytica serotype 1 (Ph1) lung infection in goats, using low numbers of bacteria and without impairing host immunity, was developed. Two trials were conducted. Results of trial 1, using 10 principals (Ph1 agar beads) and 6 controls (agar beads alone), indicated that Ph1 organisms imbedded in agar beads could survive host lung defenses for 32 days. Results of trial 2 indicated that lung immunity in the inoculated goats (principals) was high and they were more protected than controls against a transthoracic challenge of Ph1 (1.18 x 10(7) colony-forming units) injected into a lung of each goat on posttreatment day 35. When comparing challenge-exposed principals with controls, the controls developed rectal temperatures above normal for a longer time, duration of anorexia was longer, and signs of depression were seen. The controls developed large areas of consolidated lung tissue, more Ph1 isolates were recovered from nasal turbinates and lung tissue, and higher Ph1 concentrations were found in the lungs. The serum Ph1 indirect hemagglutination antibody titers in the principals of both trials increased, compared with titers in controls. Principal goats in trial 2 had higher Ph1 indirect hemagglutination antibody titers after injection of Ph1-impregnated agar beads and less severe lung lesions after challenge exposure than did controls. The small pneumonic consolidated lesions in the principals, compared with extensive lesions in controls after Ph1 challenge exposure, indicated a high degree of immunity after exposure to Ph1 organisms imbedded in agar beads.     

参麦注射液对大鼠急性肺损伤保护作用及机制的研究

试验旨在探讨参麦注射液对大鼠肺组织的保护作用及可能机制,为临床治疗急性肺损伤(acute lung injury,ALI)提供理论依据。将Wister大鼠随机分为模型组、参麦治疗组和空白对照组,每组采用经典尾静脉注射油酸方法建立急性肺损伤模型。观察肺体系数、病理切片、超微结构及肺组织匀浆中TNF-α、ICAM-1的含量变化,探讨参麦注射液对大鼠肺组织急性肺损伤的保护作用。结果发现,与模型组相比参麦治疗组可以明显改变大鼠急性肺损伤的肺体系数,减轻肺组织形态学及超微结构的病理改变,同时降低肺组织匀浆中TNF-α、ICAM-1的表达。证实参麦注射液对大鼠急性肺损伤具有保护作用,其机制可能通过降低肺组织匀浆中TNF-α、ICAM-1的含量,而发挥抗炎作用。     

A comparison of biopsy and post-mortem findings in the lungs of healthy cows

Lung biopsy in cattle for the diagnosis of lung diseases is rarely used or described. The clinical effects and the gross findings in lung and pleura after ultrasonic-guided percutaneous lung biopsy in the upright animal of healthy cattle were reported previously (Braun et al., 1999). This report describes the yield of attained lung tissue and the local tissue reaction in 60 healthy adult cows slaughtered 24 h (50 animals) and 10 days (10 animals) after invasion. The yield of lung tissue in the biopsies was high (85.5%) in 90 histologically examined biopsy specimens and judged 'good to excellent'. The local tissue reaction and the extent of haemorrhage in the lung parenchyma, both 24 h and 10 days after the biopsy, were minimal. In the latter group, the scar tissue had approximately the same dimensions as the tissue cylinders punched 10 days before. There were no local adhesions between the pleural surfaces. Previously published data concerning clinical reactions and complications, macroscopically examined local tissue bleeding and the histologically evaluated minimal local inflammatory reaction, following lung biopsy of cattle, indicate that this procedure is safe and satisfactory for sampling of accurate lung material. The results of this study concur and indicate that local complications in the animal were minimal to absent.     

NAC通过抑制大鼠肺细胞凋亡缓解镉致肺组织损伤

本试验旨在探讨N-乙酰半胱氨酸（NAC）在镉（Cd）致去卵巢大鼠肺细胞凋亡和肺组织损伤的缓解作用。40只雌性SD大鼠随机分为假手术对照（Control）组、Cd组、NAC组和NAC+Cd组，每组10只。18个月后，取大鼠肺组织，镜检观察组织HE染色与Masson染色后的病理学变化，应用火焰原子吸收光谱法检测组织中Cd的含量，应用qRT-PCR法检测组织纤维化关键蛋白Col-I和Col-III mRNA水平，并应用Western blot检测凋亡关键蛋白（Bcl-2、Bax及cleaved caspase-3）及p53的表达与Akt磷酸化。结果显示，与Control组相比，Cd组肺组织中肺泡结构消失和纤维组织增生，Cd的含量极显著增加（P<0.01），Col-I和Col-III mRNA水平、Bax/Bcl-2比值、cleaved caspase-3与p53的表达极显著上调（P<0.01），Akt磷酸化受抑制。与Cd组相比，NAC+Cd组减轻了肺组织病理变化，显著（P<0.01）降低了肺组织中Cd的含量，下调了Col-I和Col-III mRNA水平和Bax/Bcl-2比值、cleaved caspase-3与p53的表达，提高了Akt磷酸化水平。综上表明，NAC对Cd致大鼠肺组织损伤和肺细胞凋亡具有较好的缓解作用。     

Use of Electrocautery Probes in Arthroscopic Removal of Apical Sesamoid Fracture Fragments in 18 Standardbred Horses

OBJECTIVE: To develop a technique for arthroscopic excision of apical sesamoid fracture fragments using electrocautery probes. ANIMALS OR SAMPLE POPULATION: A total of 18 Standardbred horses. METHODS: Arthroscopic removal of apical sesamoid fracture fragments using both hook and loop electrocautery probes was performed on 18 Standardbred horses. The fracture fragments were approached using either an ipsilateral (3) or contralateral (15) arthroscopic triangulation technique. Distension of the metacarpo- (metatarso)-phalangeal joints was achieved using a 1.5% glycine solution. Under arthroscopic guidance, both the suspensory and intersesamoidean ligament attachments to the abaxial and the axial margins of the apical fragment were transected using a hook electrocautery probe. Subsequently, the palmar (plantar) soft tissue attachments to the apical fragment were transected with a loop electrocautery probe. Once freed of soft tissue attachments, the apical fragment was removed with Ferris-Smith intervertebral disc rongeur. The arthroscopic portals were closed in a routine manner. RESULTS: Horse ages ranged from 2 to 8 years (median, 2.5 years). Eighteen apical sesamoid fracture fragments were removed from the left (8) and right (8) hind limbs and the left (1) and right (1) forelimbs. Apical fragments occured in 15 lateral and 3 medial proximal sesamoid bones. No major complications occurred during or after surgery. The electrocautery probes permitted an easy and precise dissection of all soft tissue attachments to the apical sesamoid fracture fragments in all horses. Follow-up time was 7 to 44 months (median, 26 months); 10 of 14 horses returned to racing. Seven of nine horses that raced before surgery raced again and three of five that had not raced before surgery, raced afterwards. CONCLUSION: Arthroscopic excision of apical sesamoid fracture fragments can be accomplished arthroscopically using electrocautery probes. CLINICAL RELEVANCE: Arthroscopic electrosurgery is an efficient and safe alternative for intra-articular dissection in horses.     

COMPUTED TOMOGRAPHY OF THE LUNG OF HEALTHY SNAKES OF THE SPECIES PYTHON REGIUS, BOA CONSTRICTOR, PYTHON RETICULATUS, MORELIA VIRIDIS, EPICRATES CENCHRIA, AND MORELIA SPILOTA

Thirty-nine healthy boid snakes representing six different species ( Python regius, Boa constrictor, Python reticulatus, Morelia viridis, Epicrates cenchria , and Morelia spilota ) were examined using computed tomography (CT) to characterize the normal appearance of the respiratory tissue. Assessment was done subjectively and densitometry was performed using a defined protocol. The length of the right lung was calculated to be 11.1% of the body length, without a significant difference between species. The length of the left lung in proportion to the right was dependent on the species examined. The most developed left lung was in P. regius (81.2%), whereas in B. constrictor , the left lung was vestigial or absent (24.7%). A median attenuation of −814.6 HU and a variability of 45.9 HU were calculated for all species with no significant difference between species. Within the species, a significantly higher attenuation was found for P. regius in the dorsal and cranial aspect of the lung compared with the ventral and caudal part. In B. constrictor , the reduced left lung was significantly hyperattenuating compared with the right lung. Results of this study emphasize the value of CT and provide basic reference data for assessment of the snake lung in these species.