A comparison of immunohistochemical assays and FISH in detecting the ALK translocation in diagnostic histological and cytological lung tumor material.

Introduction:Detection of the ALK rearrangement in a solid tumor gives these patients the option of crizotinib as an oral form of anticancer treatment. The current test of choice is fluorescence in situ hybridization (FISH), but various cheaper and more convenient immunohistochemical (IHC) assays have been proposed as alternatives.Methods:Fifteen FISH-positive cases from patients, seven with data on crizotinib therapy and clinical response, were evaluated for the presence of ALK protein using three different commercially available antibodies: D5F3, using the proprietary automated system (Ventana), ALK1 (Dako), and 5A4 (Abcam). A further 14 FISH-negative and three uncertain (<15% rearrangement detected) cases were also retrieved. Of the total 32 specimens, 17 were excisions and 15 were computed tomography-guided biopsies or cytological specimens. All three antibodies were applied to all cases. Antibodies were semiquantitatively scored on intensity, and the proportion of malignant cells stained was documented. Cutoffs were set by receiver operating curve analysis for positivity to optimize correct classification.Results:All three IHC assays were 100% specific but sensitivity did vary: D5F3 86%, ALK 79%, 5A4 71%. Intensity was the most discriminating measure overall, with a combination of proportion and intensity not improving the test. No FISH-negative IHC-positive cases were seen. Two FISH-positive cases were negative with all three IHC assays. One of these had been treated with crizotinib and had failed to show clinical response. The other harbored a second driving mutation in the EGFR gene.Conclusions:IHC with all three antibodies is especially highly specific (100%) although variably sensitive (71%-86%), specifically in cases with scanty material. D5F3 assay was most sensitive in these latter cases. Occasional cases are IHC-positive but FISH-negative, suggesting either inaccuracy of one assay or occasional tumors with ALK rearrangement that do not express high levels of ALK protein

Intraoperative pleural lavage cytology is an independent prognostic indicator for staging non–small cell lung cancer

AbstractObjectivesFor patients undergoing lung resection for cancer, macroscopic evidence of metastasis is clearly associated with adverse prognosis. However, less is known about the significance of tumor cells detected by using tests such as pleural lavage cytology. To ascertain the frequency and quantify the effect of this finding on survival, we performed a prospective study of intraoperative pleural lavage cytology.MethodsPleural lavage cytology consisted of cytologic analysis of 100 mL of saline irrigated over the lung surface immediately after thoracotomy. Patients were excluded if they had an existing effusion, extreme adhesions, or lateral chest wall invasion or if resection was not performed. Survival was calculated by means of Kaplan-Meier analysis and compared by using log-rank tests. Cox regression was used to ascertain independent predictors of prognosis.ResultsFrom 1995 through 2003, we performed pleural lavage cytology on 292 patients undergoing thoracotomy for lung cancer. The mean age was 64 (SD, 10) years, and 196 (67%) patients were men. Of 292 samples, 13 (4.5%) showed evidence of malignant cells. The median time to follow-up was 15 months (interquartile range, 1-40 months), with a median survival of 49 months for patients with negative pleural lavage cytology results and 13 months for patients with positive pleural lavage cytology results (P = .002). Univariate prognostic predictors were positive pleural lavage cytology status (P = .03), stage (P = .03), adenocarcinoma (P = .06), and parietal pleural involvement (P = .01). In the final multivariate model only positive pleural lavage cytology status (P = .006) and stage (P = .03) remained significant.ConclusionsIntraoperative pleural lavage cytology is a simple addition to intrathoracic staging and an independent predictor of prognosis. Positive results potentially affect survival by upstaging patients to stage IIIB or greater

TRPV1-expressing primary afferents generate behavioral responses to pruritogens via multiple mechanisms

The mechanisms that generate itch are poorly understood at both the molecular and cellular levels despite its clinical importance. To explore the peripheral neuronal mechanisms underlying itch, we assessed the behavioral responses (scratching) produced by s.c. injection of various pruritogens in PLCβ3- or TRPV1-deficient mice. We provide evidence that at least 3 different molecular pathways contribute to the transduction of itch responses to different pruritogens: 1) histamine requires the function of both PLCβ3 and the TRPV1 channel; 2) serotonin, or a selective agonist, α-methyl-serotonin (α-Me-5-HT), requires the presence of PLCβ3 but not TRPV1, and 3) endothelin-1 (ET-1) does not require either PLCβ3 or TRPV1. To determine whether the activity of these molecules is represented in a particular subpopulation of sensory neurons, we examined the behavioral consequences of selectively eliminating 2 nonoverlapping subsets of nociceptors. The genetic ablation of MrgprD^+ neurons that represent ≈90% of cutaneous nonpeptidergic neurons did not affect the scratching responses to a number of pruritogens. In contrast, chemical ablation of the central branch of TRPV1+ nociceptors led to a significant behavioral deficit for pruritogens, including α-Me-5-HT and ET-1, that is, the TRPV1-expressing nociceptor was required, whether or not TRPV1 itself was essential. Thus, TRPV1 neurons are equipped with multiple signaling mechanisms that respond to different pruritogens. Some of these require TRPV1 function; others use alternate signal transduction pathways

Certified causes of death in patients with mesothelioma in South East England

Background: Mesothelioma is a highly fatal cancer that is caused by exposure to asbestos fibres. In many populations, the occurrence of mesothelioma is monitored with the use of mortality data from death certification. We examine certified causes of death of patients who have been diagnosed with mesothelioma, and assess the validity of death certification data as a proxy for mesothelioma incidence.Methods: We extracted mesothelioma registrations in the South East of England area between 2000 and 2004 from the Thames Cancer Registry database. We retained for analysis 2200 patients who had died at the time of analysis, after having excluded seven dead cases where the causes of death were not known to the cancer registry. The 2200 deaths were classified hierarchically to identify (1) mesothelioma deaths, (2) deaths certified as lung cancer deaths or (3) deaths from unspecified cancer, and (4) deaths from other causes.Results: 87% of the patients had mesothelioma mentioned on the death certificate. 6% had no mention of mesothelioma but included lung cancer as a cause of death. Another 6% had no mention of mesothelioma or lung cancer, but included an unspecified cancer as a cause of death. Lastly, 2% had other causes of death specified on the death certificate.Conclusion: This analysis suggests that official mortality data may underestimate the true occurrence of mesothelioma by around 10%

A 20-year review of the status and distribution of African wild dogs (Lycaon pictus) in South Africa

South Africa is one of only seven countries with a viable population of African wild dogs (Lycaon pictus). The national population in 2017 was 372 adults and yearlings and comprised three subpopulations: 1) Kruger National Park (Kruger), 2) an intensively managed metapopulation established through reintroductions into isolated, fenced reserves, and 3) a free-roaming population that occurs naturally outside protected areas. We assessed the long-term (four wild dog generations, ~20 years) trends in population size and growth rate within each of these three subpopulations. We found that Kruger supports a substantial population,which has declined over time.The metapopulation is the only subpopulation that has increased significantly over time (both in population size and number of packs), likely due to intensive conservation efforts and the reintroduction of wild dogs into 15 additional reserves since 1998. The free-roaming subpopulation has remained small but stable, even though the number of packs has declined due to anthropogenic threats. The overall national population has remained stable even though the number of packs has increased. Kruger has consistently supported the highest proportion of the national population over the last two decades. However, the contribution of the metapopulation has increased significantly over time. It is clear that despite differences in survey effort among the three subpopulations, South Africa has a small (~500) but stable population of wild dogs, with the metapopulation contribution becoming increasingly important. The circumstances in the country necessitate, and demonstrate the benefit of, intensive, adaptive management for the national population of wild dogs. While this assessment provides baseline information for the three subpopulations, wild dog conservation in South Africa would benefit greatly from equal survey effort and standardized methods to accurately assess long-term population trends.https://journals.co.za/journal/wild2pm2021Zoology and Entomolog

ASCA Observations of "Type 2" LINERs: Evidence for a Stellar Source of Ionization

We present ASCA observations of LINERs without broad H$\alpha$ emission in their optical spectra. The sample of "type 2" LINERs consists of NGC 404, 4111, 4192, 4457, and 4569. We have detected X-ray emission from all the objects except for NGC 404; among the detected objects are two so-called transition objects (NGC 4192 and NGC 4569), which have been postulated to be composite nuclei having both an HII region and a LINER component. The images of NGC 4111 and NGC 4569 in the soft (0.5-2 keV) and hard (2-7 keV) X-ray bands are extended on scales of several kpc. The X-ray spectra of NGC 4111, NGC 4457 and NGC 4569 are well fitted by a two-component model that consists of soft thermal emission with $kT\sim0.65$ keV and a hard component represented by a power law (photon index $\sim$ 2) or by thermal bremsstrahlung emission ($kT\sim$ several keV). The extended hard X-rays probably come from discrete sources, while the soft emission most likely originates from hot gas produced by active star formation in the host galaxy. We have found no clear evidence for the presence of active galactic nuclei (AGNs) in the sample. If an AGN component is the primary ionization source of the optical emission lines, then it must be heavily obscured with a column density significantly larger than $10^{23}$ cm$^{-2}$. Alternatively, the optical emission could be ionized by a population of exceptionally hot stars.Comment: 13 pages, 5 figures, emulateapj.sty, Accepted for publication in the Astrophysical Journa

One-year outcomes in a multicentre cohort study of incident rare diffuse parenchymal lung disease in children (ChILD)

We performed a prospective, observational, cohort study of children newly diagnosed with children's interstitial lung disease (ChILD), with structured follow-up at 4, 8, 12 weeks and 6 and 12 months. 127 children, median age 0.9 (IQR 0.3-7.9) years had dyspnoea (68%, 69/102), tachypnoea (75%, 77/103) and low oxygen saturation (SpO(2)) median 92% (IQR 88-96). Death (n=20, 16%) was the most common in those <6 months of age with SpO(2)<94% and developmental/surfactant disorders. We report for the first time that ChILD survivors improved multiple clinical parameters within 8-12 weeks of diagnosis. These data can inform family discussions and support clinical trial measurements

Morphometric analysis of lymphatics vessels in fibrotic human lung

In pulmonary fibrosis, the usual interstitial pneumonia (UIP) pattern is characterised by heterogeneous, patchy fibrosis, with areas of normal lung adjacent to areas of complete destruction (honeycombing) and by fibroblastic foci (FF). The NSIP pattern which is characteristic of systemic sclerosis, is characterised by a more homogeneous involvement of the lung without honeycombing and FF. Little is known on lymphatic vessels in lung fibrosis. Defective lymphatic clearance could lead to prolonged exposure to pathogenic antigens and/or pro-inflammatory/pro-fibrotic mediators. We evaluated the distribution and morphology of lymphatic vessels in lung biopsies of 6 patients with UIP, 6 NSIP and 5 controls. Consecutive sections were stained with Movat’s pentachrome and with double immunostaining for von Willebrand factor and podoplanin (D2-40). Area, perimeter and position were recorded for vessels with a diameter &gt; 5µm. We investigated separately in lintralobular, sub-pleural, and interlobular spaces. Lymphatics were consistently larger in subpleural spaces and in interlobular septa than in intralobular tissue. In the latter, the density of lymphatic vessels was significantly reduced in NSIP and in UIP (both 21±1 mm-2) compared to controls (35±4 mm-2) . In controls, 85±6% of the intralobular lymphatics were close (&lt; 100 µm) to a blood vessel, and only 5±4% were in the proximity of bronchoalveolar spaces, while in the disease groups they were less frequently perivascular (NSIP 55 ±3%, UIP 56 ±2%) and more frequently associated with the bronchoalveolar lumen (NSIP 85 ±3%, UIP 69 ±2%). By contrast, in interlobular septa, lymphatic density was significantly increased in NSIP (303±28 mm-2) and in UIP (286±124 mm-2) compared to controls (96±69 mm-2). No differences in lymphatic density was seen in subpleural spaces. Thus, our data show a marked redistribution of lymphatic vessels within the lung in pulmonary fibrosis, without noticeable differences between the NSIP and UIP patterns