Equine Airway Mast Cells are Sensitive to Cell Death Induced by Lysosomotropic Agents.

Mast cells are known for their detrimental effects in various inflammatory conditions. Regimens that induce selective mast cell apoptosis may therefore be of therapeutic significance. Earlier studies have demonstrated that murine- and human-cultured mast cells are highly sensitive to apoptosis induced by the lysosomotropic agent LeuLeuOMe (LLME). However, the efficacy of lysosomotropic agents for inducing apoptosis of in vivo-derived airway mast cells and the impact on mast cells in other species have not been assessed. Here we addressed whether lysosomotropic agents can induce cell death of equine in vivo-derived mast cells. Bronchoalveolar lavage (BAL) fluids from horses were incubated with LLME at 15-100 µm for up to 48 h. The overall cell viability was unaffected by 15 µm LLME up to 48 h, whereas a relatively modest drop in total cell counts (~30%) was seen at the highest LLME dose used. In contrast to the relatively low effect on total cell counts, LLME efficiently and dose dependently reduced the number of mast cells in BAL fluids, with an almost complete depletion (96%) of mast cells after 24 h of incubation with 100 µm LLME. A significant but less dramatic reduction (up to ~45%) of lymphocytes was also seen, whereas macrophages and neutrophils were essentially resistant. The appearance of apoptotic bodies suggested a mechanism involving apoptosis rather than necrosis. These findings suggest that equine airway mast cells are highly sensitive to lysosomotropic agents. Possibly, lysosomotropic agents could be of therapeutic value to treat disorders involving harmful accumulation of mast cells in the airways.

Death causes in breast cancer patients.

BACKGROUND: Due to improved outcomes in breast cancer (BCa), the proportion of affected women dying of other causes has increased. Thus, a better survival of BCa requires knowledge of other causes of death. MATERIALS AND METHODS: Data on the population, cancers, and causes of death were gathered from the nationwide Swedish Family-Cancer Database, enrolling ∼3.68 million Swedish women. A Cox regression model, comparing BCa patients against all other women, was applied. Cause-of-death-specific hazard ratios (HRs) were calculated for both underlying and multiple causes of death. RESULTS: Among 641 000 deaths from 1987 to 2006, 48,000 were BCa patients. For underlying causes except BCa, the highest cause-specific HRs were found for diseases of pulmonary circulation {1.51 [95% confidence interval (CI) 1.36-1.68]}, suicide [1.39 (1.19-1.63)], and heart failure [1.29 (1.22-1.37)]. For specific multiple causes, the highest ratios were found for external causes [1.86 (1.80-1.91)] and gastrointestinal disease [1.68 (1.62-1.74)]. CONCLUSIONS: Diagnosis of BCa is associated with increased risks of dying of various causes, including external causes, heart failure, diseases of pulmonary circulation, and gastrointestinal disease. The study fulfills the gap in knowledge of death causes in BCa patients and suggests to draw more attention to comorbidities.

Long term silent carriers of Streptococcus equi ssp. equi following strangles; carrier detection related to sampling site of collection and culture versus qPCR.

After strangles outbreaks, Streptococcus equi ssp. equi (S. equi) can persist in clinically normal silent carriers for months to years. Two naturally occurring outbreaks of strangles with 53 and 100% morbidity, respectively, were followed longitudinally to assess occurrence of carrier state and optimal detection methods Outbreak A involved 98 yearling warmbloods, and outbreak B 38 mature Icelandic horses. Fully recovered horses were sampled at least 6 months after index cases using nasal swabs (one sampling occasion only) nasopharyngeal lavage and guttural pouch visualisation and lavages for culture and qPCR to S. equi. Any horse with at least a single sample positive was deemed a carrier. Descriptive statistics and sensitivity and negative predictive values were calculated. Comparisons were made with McNemars and Fishers exact tests. Carrier rates in outbreak A were 3% based on culture and 15% based on qPCR and for outbreak B 13% based on culture and 37% based on qPCR. All culture positives were also qPCR positive. One carrier culture negative sampled after an additional 8 months was culture positive to S. equi, indicating that qPCR positives should be suspected to carry live bacteria. Findings indicate that reliance on guttural pouch sampling and appearance does not capture all silent carriers. All culture positives were identified by qPCR and even horses positive by qPCR but culture negative should be suspected carriers of live bacteria.

Partial divergence of cytokine mRNA expression in bronchial tissues compared to bronchoalveolar lavage cells in horses with recurrent airway obstruction.

The aim of this study was to investigate mRNA levels of cytokines in bronchial epithelium in horses with recurrent airway obstruction (RAO) during acute crisis and remission. Additionally, cytokine mRNA levels in endobronchial biopsies and bronchoalveolar lavage (BAL) cells were compared. Seven RAO horses were examined while in respiratory crisis following provocation and again while in remission after 2 months on pasture, during which time six healthy horses on pasture were also examined. Quantitative real-time PCR (RT-PCR) was used to assess mRNA expression for cytokines IL-5, IL-6, IL-8, IL-10, IL-17 and transforming growth factor beta1 (TGF-beta1) in endobronchial biopsies and bronchoalveolar lavage. Expression of IL-8 mRNA was significantly upregulated during crisis in both endobronchial biopsies and BAL cells (p=0.036), while there was a similar trend for upregulation of IL-10 mRNA only in BAL cells that approached significance (p=0.059). Moreover, during crisis the expression of IL-8 mRNA in BAL cells was positively correlated to relative IL-6 mRNA expression (r(s)=0.971, p=0.001) and bronchial epithelial expression of IL-10 and TGF-beta1 mRNA were positively correlated (r(s)=0.943, p=0.005). In comparing the relationship of mRNA expression in BAL to biopsy in individual RAO horses, there was a positive correlation with IL-6 to IL-8 mRNA expression in BAL during respiratory crisis (r(s)=0.971, p=0.001) that also correlated positively with IL-8 expression in biopsies on pasture (r(s)=0.986, p<0.0001 for both). Regarding RAO horses at pasture versus controls neither the cytokine mRNA levels in endobronchial biopsy nor in BAL cells differed significantly. These results further support previous findings that IL-8 mRNA in both BAL cells and bronchial epithelium is upregulated in RAO horses during crisis. However, apart from IL-8, it appears that expression of other cytokines, including IL-5, IL-6, IL-10, IL-17 and TGF-beta1 in bronchial epithelium does not necessarily mirror cytokine expression in BAL cells in individual horses with RAO. Accordingly, examination of markers of inflammation in endobronchial tissue provides complementary but not necessarily identical information to that obtained in BAL cells. Given the potential for repeated sampling over time bronchial biopsy can serve as an invaluable additional tool for investigation of time-dependent changes in inflammatory process in this animal model of asthma.

Clinical alterations and mRNA levels of IL-4 and IL-5 in bronchoalveolar cells of horses with transient pulmonary eosinophilia.

The aim of this study was to assess clinical signs and altered pulmonary cell expression of cytokines related to eosinophil kinetics in horses with pulmonary eosinophilia. Pulmonary eosinophilia was detected by bronchoalveolar lavage (BAL) in a group of standardbreds in training. Horses had detailed clinical examination, bronchoscopy, endobronchial biopsy and BAL on three occasions at approximately 6 month intervals. During the second sampling period BAL eosinophils were significantly elevated (p>0.010), with five horses having from 5% to 37% eosinophils in BAL. Neither detailed clinical examination parameters nor gene expression of IL-4 and IL-5 mRNA (real-time-PCR) were associated with BAL eosinophilia. Pulmonary eosinophilia abated without treatment apart from deworming. It appears that pronounced lung eosinophilia in horses can be transient, abate without specific treatment, and in this instance, lack correlation to upregulation of expression of either IL-4 or IL-5.

Decreased Clinical Severity of Strangles in Weanlings Associated with Restricted Seroconversion to Optimized Streptococcus equi ssp equi Assays.

BACKGROUND: Streptococcus equi ssp. equi causes characteristic clinical signs that are most severe in young horses, including fever, purulent nasal discharge, and lymph node abscessation in the head region. HYPOTHESIS/OBJECTIVES: Clinical, serologic, and microbiologic factors related to unexpectedly mild disease severity in a natural outbreak of strangles in immunologically naïve weanlings were investigated. ANIMALS: One-hundred and twelve warmblood weanlings. METHODS: Prospective longitudinal observational study of a natural outbreak of strangles. The entire cohort was examined at the peak of the outbreak by deep nasal swabs for culture and quantitative PCR (qPCR) for the presence of S. equi and clinically and serologically in a sequential manner by an optimized ELISA from the index case throughout the outbreak until resolution. Descriptive statistics were calculated and comparisons made using a nondirectional Wilcoxon signed-rank test. RESULTS: Outbreak morbidity was 53%, with 9 of 14 horses culture positive and 26 of 53 horses qPCR positive for S. equi lacking clinical signs characteristic of strangles. By resolution, 91 of 112 had seroconverted to Antigen A by ELISA but seroconversion to antigen C (part of the SeM protein) was minimal. Sequencing of the isolates detected no alterations in the SeM protein, but identified a 61 bp deletion in the gene SEQ_0402. CONCLUSIONS AND CLINICAL IMPORTANCE: Absence of clinical signs alone in naïve horses may be an insufficient criterion to release horses from strangles quarantine measures. Restricted seroconversion to antigen C may have been associated with decreased clinical severity. The role of a minor gene deletion in SEQ_0402 in the virulence of S. equi warrants further investigation.

Metastatic sites and survival in lung cancer.

OBJECTIVES: Population-based data on metastatic sites and survival in site-specific metastases are lacking for lung cancer and for any cancer because most cancer registries do not record metastases. This study uses a novel population-based approach to identify metastases from both death certificates and national inpatient data to describe metastatic pathways in lung cancer patients. MATERIALS AND METHODS: 17,431 deceased lung cancer patients diagnosed 2002-2010 were identified from the nationwide Swedish Cancer Registry, which is based on compulsory reports. The influence of age at diagnosis, sex, and histological subtype on metastatic spread was investigated. Survival in metastatic lung cancer was assessed by histology and metastatic site. RESULTS: The most frequent metastatic sites were the nervous system, bone, liver, respiratory system, and adrenal gland. Liver (35%) and nervous system (47%) metastases were common in patients with metastases from small cell lung cancer, and bone (39%) and respiratory system (22%) metastases in adenocarcinoma. Women (43% vs. 35%) and younger patients had more metastases to the nervous system. Median survival after diagnosis was 13 months for non-metastatic and five months for metastatic lung cancer. In this novel data, liver metastases conferred the worst prognosis (three months), especially for large cell histology. Bone metastases also featured poor survival, whereas survival in respiratory and nervous system metastases was better. CONCLUSION: Metastatic sites and survival in metastatic lung cancer is influenced by sex, histological subtype, and age at diagnosis. Liver and bone metastases signal poor survival, compared with nervous system metastases.

The clinical significance of Nicoletella semolina in horses with respiratory disorders and a screening of the bacterial flora in the airways of horses.

Nicoletella semolina, a member of the family Pasteurellaceae, can be isolated from the airways of horses with respiratory disorders. However, its role as a potential or opportunistic pathogen is not clear nor is its presence as part of the normal flora. We therefore investigated the presence and bacterial load of N. semolina in healthy and diseased horses. Samples from a healthy control group were compared with samples from the routine analysis of horses with a clinical history of respiratory disorders. A total of 1770 nose swabs and 1132 tracheal aspirate samples were analysed and subjected to conventional bacteriological examination. N. semolina was isolated from 12 (6%) of 207 nose samples from the healthy control group and from 42 (3%) of 1563 samples from horses with respiratory disorders. In tracheal aspirate, N. semolina was isolated from 7 (3%) of 211 samples from the control group and 49 (5%) of 921 samples from horses with respiratory disorders. Other bacteria were also isolated in laboratory analyses, the most commonly isolated bacterium in both the control group and the respiratory disorders group being Streptococcus equi subsp. zooepidemicus. It was isolated in 21% of tracheal aspirate from the control group and 33% of those from horses with respiratory disorders. In conclusion, N. semolina is not a primary pathogenic bacterium, as it was isolated at similar frequencies in horses with respiratory disorders and those in the healthy control group.

Time trends in survival from cancer of unknown primary: small steps forward.

BACKGROUND: Cancer of unknown primary (CUP) is a fatal cancer for which incidence trends have changed but detailed survival trends remain unexplored. These could point out successful diagnostic and therapeutic approaches. We investigate survival trends in CUP according to histology, locations of metastases and site-specific causes of death. PATIENTS AND METHODS: A total of 20,523 CUP patients with nodal and extranodal metastases were identified from the Swedish Cancer Registry. Hazard ratios (HRs) were estimated, comparing three different time periods (1987-1993, 1994-2000 and 2001-2008) with respect to histological subtype, CUP location and the cause of death. RESULTS: Survival for patients with CUP increased over the study period (HR=0.91 [95% confidence interval (CI): 0.78-0.84], p<0.001 for trend). Adenocarcinoma was the only histology associated with increased survival (0.78 [0.74-0.82], p<0.001 for trend). Survival was improved most clearly for CUP of the pelvis (0.55 [0.36-0.83]), peritoneum (0.58 [0.53-0.65]) and nervous system (0.46 [0.29-0.72]). Survival improved substantially in patients with ovarian (0.57 [0.46-0.70]), peritoneal (0.39 [0.24-0.65]) and biliary system cancers (0.67 [0.52-0.87]). Kaplan-Meier curves showed significant survival gains for all CUP and adenocarcinoma patients (p<0.001). CONCLUSIONS: Over time, survival for patients with CUP increased for adenocarcinoma and for CUP of the pelvis, peritoneum and nervous system. Survival trends in CUP may be related to (1) similar trends in other common metastatic tumours, particularly pancreatic and hepatobiliary cancers, which are common 'hidden' primaries for CUP, (2) earlier detection and (3) advances in the management of metastatic cancers. The improvement in survival at specific locations suggests true therapeutic gains.

Selection on flowering time and floral display in an alpine and a lowland population of Arabidopsis lyrata.

To determine whether population differentiation in flowering time is consistent with differences in current selection, we quantified phenotypic selection acting through female reproductive success on flowering phenology and floral display in two Scandinavian populations of the outcrossing, perennial herb Arabidopsis lyrata in two years. One population was located in an alpine environment strongly affected by grazing, whereas the other was close to sea level and only moderately affected by herbivory. Multiple regression models indicated directional selection for early end of flowering in one year in the lowland population, and directional selection for early start of flowering in one year in the alpine population. As expected, there was selection for more inflorescences in the lowland population. However, in the alpine population, plants with many inflorescences were selectively grazed and the number of inflorescences produced was negatively related to female fitness in one year and not significantly related to female fitness in the second year. The results are consistent with the hypothesis that genetic differentiation in flowering phenology between the study populations is adaptive, and indicate that interactions with selective grazers may strongly influence selection on floral display in A. lyrata.