[What Do (General/Abdominal) Surgeons Need to Know About Project Management?] / Was muss der (Allgemein-/Viszeral-)Chirurg von Projektmanagement wissen?

Clinical departments are constantly faced with complex and novel tasks, which have to be managed with suitable approaches. This article describes selected aspects of project management in this context. In particular, it concentrates on the advantages and limitations of project management methods that have been established in industrial companies when used in a clinical setting. Also it compares the daily reality in clinical departments and industrial companies and highlights possible associations between surgery and project management. Besides aspects such as project planning, role definitions within projects and their specific casting in clinics, the article also demonstrates key success factors for an effective implementation of projects and a general introduction to project management in clinics. The article combines theoretical approaches with practical experience and thought-provoking impulses.

Hsa-mir-124-3 CpG island methylation is associated with advanced tumours and disease recurrence of patients with clear cell renal cell carcinoma.

BACKGROUND: Whether methylation of the microRNA (mir)-124-3 CpG island is of relevance for the clinical course of a solid cancer and whether it shows association with clinicopathology or survival of patients with renal cell cancer (RCC) is not known as yet. METHODS: In a cross-sectional study, relative methylation of mir-124-3 was measured in 111 RCC samples and 77 paired normal appearing tissues using quantitative methyl-specific PCR. Results were statistically compared with tumour histology, clinicopathological parameters and disease recurrence. RESULTS: We found tumour-specific hypermethylation of mir-124-3 in samples of RCCs with clear cell histology (ccRCC) compared with paired normal appearing tissues (P<0.0001). Methylation was significantly increased in tumours with state of advanced disease (P<0.0001). Higher relative methylation was associated with worse recurrence-free survival in both univariate (hazard ratio=9.37; P=0.0005) as well as bivariate Cox regression analyses considering age, sex, diameter of tumours and state of advanced disease, metastasis and lymph node metastases as covariates (hazard ratios=5.9-18.2; P-values of 0.0003-0.008). CONCLUSION: We identified mir-124-3 CpG islands (CGI) methylation as a relevant epigenetic mark for ccRCC thus underlining the need for functional studies of potentially affected signalling pathways in kidney tumour models. Methylation of mir-124-3 is suggested as an independent prognosticator for ccRCC.

The cat as a model for human obesity: insights into depot-specific inflammation associated with feline obesity.

According to human research, the location of fat accumulation seems to play an important role in the induction of obesity-related inflammatory complications. To evaluate whether an inflammatory response to obesity depends on adipose tissue location, adipokine gene expression, presence of immune cells and adipocyte cell size of subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) were compared between lean and obese cats. Additionally, the present study proposes the cat as a model for human obesity and highlights the importance of animal models for human research. A total of ten chronically obese and ten lean control cats were included in the present study. Body weight, body condition score and body composition were determined. T-lymphocyte, B-lymphocyte, macrophage concentrations and adipocyte cell size were measured in adipose tissue at different locations. Serum leptin concentration and the mRNA expression of leptin and adiponectin, monocyte chemoattractant protein-1, chemoligand-5, IL-8, TNF-alpha, interferon-gamma, IL-6 and IL-10 were measured in blood and adipose tissues (abdominal and inguinal SAT, and omental, bladder and renal VAT). Feline obesity was characterised by increased adipocyte cell size and altered adipokine gene expression, in favour of pro-inflammatory cytokines and chemokines. Consequently, concentration of T-lymphocytes was increased in the adipose tissue of obese cats. Alteration of adipose tissue was location dependent in both lean and obese cats. Moreover, the observed changes were more prominent in SAT compared with VAT.

Oxathiazinane derivatives display both antineoplastic and antibacterial activity: a structure activity study.

PURPOSE: The Oxathiazinane substance class is characterized by a high diversity of chemical structures yet to be fully investigated. Our research group recently proved that the 1.4.5-oxathiazine-4.4-dioxide, known as substance GP-2250, possesses antineoplastic properties as shown on pancreatic carcinoma. This current study aims to gain insights into the structure and activity relationship of a series of different Oxathiazinanes regarding their antineoplastic activity and the potential correlation with antibacterial activity. We investigated the newly synthesized Oxathiazinane derivatives: 2255, 2256, 2287, 2289, 2293 and 2296 in comparison to GP-2250. METHODS: The antineoplastic effect was evaluated in different cancer entities (breast, skin, pancreas and colon cancer cell lines) by viability, proliferation, and cell migration assays in vitro. Disc diffusion tests were performed on various bacteria strains to examine the antibacterial potential. Additionally, reactive oxygen species (ROS) assays were conducted to investigate mechanistic aspects. RESULTS: The substances GP-2250, 2293, 2289 and 2296 not only showed antineoplastic activity in four different cancer entities but also antibacterial effects, as tested on multiple bacteria strains including MRSA (Methicillin-resistant Staphylococcus aureus). Furthermore, these substances also induced high ROS levels up to 110% in the treated cancer cell lines compared to untreated control cells. These results indicate a correlation between an antineoplastic capacity and antibacterial properties of these derivatives. Both activities appear to be ROS driven. The Oxathiazinane derivatives 2255, 2256 and 2287 lacked both, antineoplastic and antibacterial activity. CONCLUSION: Thus, a comparable structure activity relationship became apparent for both the antineoplastic and antibacterial activity.

An urban perinatal health programme of strategies to improve perinatal health.

Promotion of a healthy pregnancy is a top priority of the health care policy in many European countries. Perinatal mortality is an important indicator of the success of this policy. Recently, it was shown that the Netherlands has relatively high perinatal death rates when compared to other European countries. This is in particular true for large cities where perinatal mortality rates are 20-50% higher than elsewhere. Consequently in the Netherlands, there is heated debate on how to tackle these problems. Without the introduction of measures throughout the entire perinatal health care chain, pregnancy outcomes are difficult to improve. With the support of health care professionals, the City of Rotterdam and the Erasmus University Medical Centre have taken the initiative to develop an urban perinatal health programme called 'Ready for a Baby'. The main objective of this municipal 10-year programme is to improve perinatal health and to reduce perinatal mortality in all districts to at least the current national average of l0 per 1000. Key elements are the understanding of the mechanisms of the large health differences between women living in deprived and non-deprived urban areas. Risk guided care, orientation towards shared-care and improvement of collaborations between health care professionals shapes the interventions that are being developed. Major attention is given to the development of methods to improve risk-selection before and during pregnancy and methods to reach low-educated and immigrant groups.

[Immunotherapy and tyrosine kinase inhibitors in first-line treatment of metastatic renal cell carcinoma-Which strategy when?] / Immuntherapie und Tyrosinkinaseinhibitoren beim metastasierten Nierenzellkarzinom in der First-line-Therapie ­ Wann welche Strategie?

Immunotherapies with checkpoint inhibitors have led to a paradigm shift in metastatic renal cell carcinoma (mRCC) as they established a new standard in first-line treatment. In addition to the established monotherapy with tyrosine kinase inhibitors, the spectrum of first-line options has now become wider. Based on data from studies and current guideline recommendations, this article discusses possible factors for individual strategies in first-line treatment of mRCC. For this decision, the leading criterion is the patient's risk score. In addition, the efficacy and tolerability of the substances, tumor burden, patient age and preferences as well as considerations about sequence treatment can support the decision. Real-world data for the new combination treatment, biomarkers for personalized medicine as well as studies on optimal sequence treatment for mRCC are needed.

Comparison of the value of measurement of serum galactomannan and Aspergillus-specific antibodies in the diagnosis of canine sino-nasal aspergillosis.

Serology is currently used for the diagnosis of canine sino-nasal aspergillosis (SNA). However, the accuracy of serological testing using commercially available, standardized purified antigen preparations of Aspergillus (CAPurAspAg) has only been poorly documented. The aim of the present study was to assess the diagnostic value of an agar-gel double immunodiffusion (AGDD) test and an anti-Aspergillus IgG ELISA, using CAPurAspAg and the commercially available Platelia test for the detection of serum galactomannan. Sera from 17 dogs with SNA, 18 dogs with a nasal tumour (NT), 11 dogs with lymphoplasmacytic rhinitis (LPR) and 33 control dogs were tested with the 3 methods. AGDD result was positive in 76.5% of dogs with SNA, whereas all sera from dogs with non-fungal nasal disease and control dogs were negative. A positive IgG ELISA result was obtained in 88% of dogs with SNA and in 18% of dogs with LPR. All patients with NT and control dogs had a negative IgG ELISA result. The Platelia test was positive in 24% of dogs with SNA, 11% of dogs with NT, 9% of dogs with LPR and 24% of control dogs. The results of this study suggest that (1) the detection of serum Aspergillus-specific antibodies with AGDD or ELISA, using CAPurAspAg, provides excellent specificity and good sensitivity, (2) the specificity is higher for AGDD (100%) than for ELISA (96.8%) while sensitivity is higher for ELISA (88.2%) than for AGDD (76.5%) and (3) serum galactomannan quantification with the Plateliat test is unreliable for the diagnosis of canine SNA.

[Methylation of the RASSF1A tumor suppressor gene promoter. Risk factor for carcinogenesis of urological tumors]. / Die Methylierung des RASSF1A-Tumorsuppressorgenpromotors. Risikofaktor für die Karzinogenese urologischer Tumoren.

Molecular targets of known risk factors for the development of urological tumors, such as age, smoking, and adiposity, have not yet been elucidated. Hypermethylation of CpG islands in promoters can lead to silencing of gene expression and has frequently been detected in tumors. Age-dependent accumulation of methylation of gene promoters has been observed in various normal tissues and is discussed as a common risk factor for carcinogenesis.Here we describe the RASSF1A tumor suppressor gene as exhibiting an age-dependent promoter methylation in normal kidney tissue, which is additionally affected by the risk factors of anthracosis and adiposity. Furthermore, we found significantly increased methylation of the RASSF1A promoter when comparing peripheral versus central zone prostatic tissue samples.Preliminary expression analysis indicates that RASSF1A could be involved in early tumorigenesis. Our results support the hypothesis that age and other lifestyle-dependent factors may influence promoter methylation of specific genes, possibly serving as future individual tumor risk markers.

Assessment of Lymphoid Molecular Clonality in Canine Thymoma.

The aim of this study was to document the molecular clonality of lymphoid cells in canine thymoma using polymerase chain reaction for antigen receptor rearrangement (PARR). Fifteen formalin-fixed and paraffin wax-embedded samples of canine thymoma were analyzed for T- and B-cell receptor clonality. Two of these 15 cases were excluded from the study due to insufficient DNA concentration. Twelve of the 13 remaining samples (92.3%) showed a polyclonal lymphoid component and in one case the lymphoid component was monoclonal (T-cell clonality). PARR could therefore be a useful tool for differentiating canine thymoma from canine mediastinal lymphoma.

Detection of specific bacterial agents by quantitative PCR assays in the bronchoalveolar lavage fluid of dogs with eosinophilic bronchopneumopathy vs. dogs with chronic bronchitis and healthy dogs.

In humans, Mycoplasma pneumoniae and Bordetella pertussis infections are suggested to trigger or exacerbate asthma. Whether Mycoplasma or Bordetella are associated with chronic inflammatory bronchial diseases in dogs has not been investigated. The aim of this study was to assess detection rates of Mycoplasma canis (M. canis), M. cynos and Bordetella bronchiseptica (Bb), in dogs with eosinophilic bronchopneumopathy (EBP) and chronic bronchitis (CB), compared with healthy dogs. Specific quantitative PCR (qPCR) analysis for M. canis, M. cynos and Bb were retrospectively performed on bronchoalveolar lavage fluid (BALF) collected from 24 dogs with EBP, 21 dogs with CB and 15 healthy dogs. Possible associations between qPCR results and age, BALF cytology or clinical severity scores (CSS) in dogs with EBP were investigated. There was no difference in M. canis, M. cynos and Bb detection rates in dogs with EBP (n=6, n=2 and n=6, respectively) and dogs with CB (n=2, n=2 and n=2, respectively) compared with control dogs (n=4, n=2 and n=2, respectively). In dogs with EBP, the proportion that were qPCR-positive for Bb was higher in dogs with higher CSS (P=0.014) and BALF from Bb-positive dogs had higher percentage of neutrophils (P<0.001). Among dogs that were qPCR-positive for Bb, moderate to high loads were only detected in dogs with EBP. M. canis and M. cynos detection was not associated with EBP or CB; higher Bb loads were only present in dogs with EBP and high CSS. A possible cause and effect relationship between Bb infection or load and EBP remains unclear and requires further investigation.

Angiostrongylosis in dogs with negative fecal and in-clinic rapid serological tests: 7 Cases (2013-2017).

BACKGROUND: Angiostrongylosis is considered as emerging disease in dogs in Belgium. Detection of first-stage larvae in feces using the Baermann method has an imperfect sensitivity. OBJECTIVES: Investigation of efficacy of noninvasive blood and fecal diagnostic tests in comparison with PCR on bronchoalveolar lavage (BAL) material in a small series of coughing or dyspnoeic dogs naturally infected with Angiostrongylus vasorum. ANIMALS: Seven dogs with angiostrongylosis. METHODS: Retrospective study. Dogs with cough, exercise intolerance and dyspnea of 2- to 8-week duration. Diagnostic methods used included Baermann analysis, AngioDetect rapid assay, ELISAs for detection of circulating antigen and specific antibodies and qPCR on BAL material. RESULTS: Baermann analysis, AngioDetect rapid assay, antigen ELISA, antibody ELISA, and qPCR on BAL material were positive in 3/7, 2/7, 3/6, 6/6, and 7/7 dogs, respectively. ELISA for antibodies or qPCR on BAL material were essential for definitive diagnosis in 3 dogs. Relative sensitivities of AngioDetect rapid assay, Baermann analysis, and ELISA for antigen detection were lower than 50% compared with ELISA for antibodies or qPCR on BAL material. CONCLUSION AND CLINICAL IMPORTANCE: In this small clinical series, Baermann analysis and AngioDetect rapid assay failed to confirm the diagnosis in some dogs. Therefore, ELISA for antibody detection and qPCR on BAL material should strongly be considered in clinically suspected dogs when antigen detection methods (AngioDetect or ELISA) and Baermann analysis are negative.

Distinct tissue cytokine and chemokine mRNA expression in canine sino-nasal aspergillosis and idiopathic lymphoplasmacytic rhinitis.

Idiopathic lymphoplasmacytic rhinitis (LPR) and sino-nasal aspergillosis (SNA) are among the most common causes of nasal discharge in dogs. The pathogenesis of both diseases is poorly understood. Some have proposed that LPR is a chronic inflammatory response to an inhaled irritant, pollutant or allergen, but others suggest that most cases of LPR constitute undiagnosed cases of SNA. Local immune dysfunction is thought to permit opportunist infection in canine SNA. This study investigates the nature of the local tissue immune response mounted in canine LPR and SNA in order to determine whether these diseases have similar or distinct pathogenesis. Quantitative reverse transcriptase polymerase chain reaction was carried out on RNA isolated from nasal biopsies from diseased and control dogs, using specific assays designed to amplify messenger RNA (mRNA), encoding a panel of cytokines and chemokines. SNA was associated with significantly increased expression of mRNA encoding interleukin (IL)-6, IL-8, IL-10, IL-12p19, IL-12p35, IL-12p40, IL-18, IFN-gamma, TNF-alpha, TGF-beta, eotaxin-2 and all four monocyte chemoattractant proteins (MCPs) relative to controls. LPR was associated with significantly increased expression of mRNA encoding IL-5, IL-8, IL-10, IL-12p19, IL-12p40, IL-18, TNF-alpha, TGF-beta, MCP-2 and MCP-3 relative to controls. There was significantly more expression of mRNA encoding IL-6, IL-8, IL-10, IL-12p35, IL-12p40, IL-18, IFN-gamma, TNF-alpha, TGF-beta and all MCPs, and significantly less expression of IL-5 in dogs with SNA than in dogs with LPR. Thus, the profile of cytokine and chemokine gene expression in the nasal mucosa is different in dogs with LPR when compared to dogs with SNA. A partial Th2 immune response appears to be mounted in the nasal mucosa of dogs with LPR, whereas the mucosal immune response in canine SNA is of the Th1 type. Increase in IL-10 and TGF-beta transcripts in dogs with SNA is thought to be implicated in the failure to clear the Aspergillus infection. These results constitute the first evidence that the pathogenesis of canine LPR and SNA is distinct.

Dynamic regulation of activated leukocyte cell adhesion molecule-mediated homotypic cell adhesion through the actin cytoskeleton.

Restricted expression of activated leukocyte cell adhesion molecule (ALCAM) by hematopoietic cells suggests an important role in the immune system and hematopoiesis. To get insight into the mechanisms that control ALCAM-mediated adhesion we have investigated homotypic ALCAM-ALCAM interactions. Here, we demonstrate that the cytoskeleton regulates ALCAM-mediated cell adhesion because inhibition of actin polymerization by cytochalasin D (CytD) strongly induces homotypic ALCAM-ALCAM interactions. This induction of cell adhesion is likely due to clustering of ALCAM at the cell surface, which is observed after CytD treatment. Single-particle tracking demonstrated that the lateral mobility of ALCAM in the cell membrane is increased 30-fold after CytD treatment. In contrast, both surface distribution and adhesion of a glycosylphosphatidylinositol (GPI)-anchored ALCAM mutant are insensitive to CytD, despite the increase in lateral mobility of GPI-ALCAM upon CytD treatment. This demonstrates that clustering of ALCAM is essential for cell adhesion, whereas enhanced diffusion of ALCAM alone is not sufficient for cluster formation. In addition, upon ligand binding, both free diffusion and the freely dragged distance of wild-type ALCAM, but not of GPI-ALCAM, are reduced over time, suggesting strengthening of the cytoskeleton linkage. From these findings we conclude that activation of ALCAM-mediated adhesion is dynamically regulated through actin cytoskeleton-dependent clustering.

Effect of culturally competent educational films about prenatal screening on informed decision making of pregnant women in the Netherlands.

OBJECTIVE: To evaluate the effect of a culturally competent educational film (CCEF) on informed decision making (IDM) regarding prenatal screening (PS) in a study population consisting of multicultural pregnant women. METHODS: A cross-sectional study with 262 women in the control group and 117 in the intervention group. All counselled participants received a self-report questionnaire to obtain data on IDM and only the intervention group received the CCEF. Twenty two percent of the study population had an ethnic minority background and 52% had a low or medium educational level. RESULTS: After exposure to the CCEF, knowledge about the Fetal Anomaly Scan (FAS) was significantly increased in ethnic minority women and in 'medium' and 'highly' educated women. Among women in the intervention group who had the intention to participate in FAS, there was an increase of 11% in IDM and a decrease of 12% in uninformed decision making. CONCLUSION: CCEF leads to a significant increase in the level of knowledge in medium and highly educated groups as well as non-western ethnic minority groups. The increase in IDM among intentional participants in the FAS is promising as well. CCEF's are a valuable complement to counseling about PS.

[MRI/TRUS fusion-guided prostate biopsy : Value in the context of focal therapy]. / MRT/TRUS-fusionierte Biopsiesysteme : Stellenwert bei der fokalen Therapie des Prostatakarzinoms.

BACKGROUND: Several systems for MRI/TRUS fusion-guided biopsy of the prostate are commercially available. Many studies have shown superiority of fusion systems for tumor detection and diagnostic quality compared to random biopsy. The benefit of fusion systems in focal therapy of prostate cancer (PC) is less clear. OBJECTIVES: Critical considerations of fusion systems for planning and monitoring of focal therapy of PC were investigated. MATERIALS AND METHODS: A systematic literature review of available fusion systems for the period 2013-5/2016 was performed. A checklist of technical details, suitability for special anatomic situations and suitability for focal therapy was established by the German working group for focal therapy (Arbeitskreis fokale und Mikrotherapie). RESULTS: Eight fusion systems were considered (Artemis™, BioJet, BiopSee®, iSR´obot™ Mona Lisa, Hitachi HI-RVS, UroNav and Urostation®). Differences were found for biopsy mode (transrectal, perineal, both), fusion mode (elastic or rigid), navigation (image-based, electromagnetic sensor-based or mechanical sensor-based) and space requirements. DISCUSSION: Several consensus groups recommend fusion systems for focal therapy. Useful features are "needle tracking" and compatibility between fusion system and treatment device (available for Artemis™, BiopSee® and Urostation® with Focal One®; BiopSee®, Hitachi HI-RVS with NanoKnife®; BioJet, BiopSee® with cryoablation, brachytherapy). CONCLUSIONS: There are a few studies for treatment planning. However, studies on treatment monitoring after focal therapy are missing.

Real-time RT-PCR quantification of mRNA encoding cytokines, CC chemokines and CCR3 in bronchial biopsies from dogs with eosinophilic bronchopneumopathy.

Idiopathic canine eosinophilic bronchopneumopathy (EBP) is a disease characterized by eosinophilic infiltration of the pulmonary interstitium and bronchial mucosa, a cause for which has not yet been discovered. A recent study, examining the relative proportion of various lymphocyte cell subsets within bronchoalveolar lavage fluid from dogs with EBP, has shown a selective increase in CD4(+) T-cells and a selective decrease in CD8(+) T-cells, suggesting that a similar Th2 immune response might occur in EBP. The aim of the present study was to determine the profile of cytokine, chemokine and CC chemokine receptor 3 (CCR3) messenger RNA (mRNA) expression in bronchial tissue from dogs with EBP. Real-time RT-PCR assays were used for the quantification of mRNA encoding for a panel of cytokines, CC chemokines and CCR3 in perendoscopic bronchial biopsies from eight dogs with EBP and seven age-matched control dogs. Messenger RNA transcribed from the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase was used for normalisation of the threshold cycle in order to determine the relative copy numbers of the transcripts. No significant difference in the expression of any cytokine, MCP-1, -2, -4 and CCR3 was found between control and EBP dogs. The expression of transcript for MCP-3, eotaxin-2 and -3 was significantly greater in bronchial biopsies from dogs with EBP than in samples from control dogs while there was significantly less mRNA encoding RANTES in the mucosa of dogs with EBP. In conclusion, the cytokine mRNA expression profile in perendoscopic bronchial biopsies is similar in dogs with EBP and dogs without respiratory disease. Further studies on the quantification of mRNA encoding cytokines in isolated T lymphocytes from bronchoalveolar lavage fluid or bronchial biopsies are needed before any conclusion on the cytokine profile in canine EBP can be drawn. Eotaxin-2, -3 and MCP-3 appear to be implicated in the pathogenesis of the disease.

The effects of morphine on human 5-HT3A receptors.

5-HT3 receptors are ligand-gated ion channels that are involved in the modulation of emesis and pain. In this study, we investigated whether the opioid analgesic, morphine, exerts specific effects on human 5-HT3 receptors. Whole-cell patches from HEK-293 cells stably transfected with the human 5-HT3A receptor cDNA were used to determine the effects of morphine on the 5-HT-induced currents using the patch clamp technique. At negative membrane potentials, 5-HT induced inward currents in a concentration-dependent manner. The 5-HT3 receptor antagonist, ondansetron, (0.3 nM) reversibly inhibited the 5-HT-induced signals. Morphine reversibly suppressed 5-HT-induced peak currents as a function of concentration (IC50 = 1.1 microM, Hill coefficient = 1.2). The block by morphine decreased with increasing 5-HT concentrations, suggesting a competitive effect. In addition, the activation, as well as the inactivation, kinetics of the currents were significantly slowed in the presence of morphine. The morphine antagonist, naloxone, also inhibited 5-HT-induced currents (e.g., at 3 microM by 17%). The effects of morphine and naloxone were not additive. The potency of morphine and the competitivity of the blocking effect points to a specific mechanism at a receptor site rather than an unspecific membrane effect.

Palladium-gold catalyst for the electrochemical reduction of CO2 to C1-C5 hydrocarbons.

Copper is a unique electrocatalyst for CO2 reduction, since it is one of the few catalysts able to produce methane, ethylene and ethane from CO2 with decent faradaic efficiencies. Here we report on the design and synthesis of a new non-copper-containing catalyst able to reduce CO2 to C1 to C5 hydrocarbons. This catalyst was designed by combining a metal that binds CO strongly, Pd, with a metal that binds CO weakly, Au, in an attempt to tune the binding energy of CO. We show that a mixture of C1-C5 hydrocarbons and soluble products are produced from an onset potential of -0.8 VRHE. We propose that the higher hydrocarbons are formed via a polymerization of -CH2 groups adsorbed on the catalyst surface.

Quantitative PCR and Cytology of Bronchoalveolar Lavage Fluid in Dogs with Bordetella bronchiseptica Infection.

BACKGROUND: The use of quantitative PCR (qPCR) for detection of Bordetella bronchiseptica in bronchoalveolar lavage fluid (BALF) and demonstration of bacteria adhering to ciliated epithelial cells in BALF or bronchial brushing fluid (BBF) has not been assessed in a series of affected dogs. Coinfections can worsen the clinical severity in bordetellosis, but the specific association with Mycoplasma cynos has not been evaluated. OBJECTIVES: To assess the utility of culture, qPCR and cytologic examination of cytospin preparations in the diagnosis of bordetellosis in dogs and the influence of coinfection by M. cynos on disease severity. ANIMALS: Twenty-four referred dogs with B. bronchiseptica infection and 10 healthy dogs. METHODS: Retrospective case series. qPCR (B. bronchiseptica and M. cynos) and culture results from BALF were recorded. Cytospin preparations from BALF and BBF were reviewed. qPCR on BALF from 10 healthy dogs were used as negative control. RESULTS: The BALF culture and qPCR detected B. bronchiseptica in 14/24 and 18/18 dogs, respectively. Coccobacilli were found adhering to ciliated epithelial cells in 20 of the 21 BALF cytologic preparations where epithelial cells were found, and 2/3 BBF cytologic preparations. Quantitative PCR detected a low level of B. bronchiseptica in one healthy dog. The frequency of detection of M. cynos was not significantly different in B. bronchiseptica (9/17 dogs) compared with healthy dogs (2/10 dogs) (P = .09). CONCLUSION AND CLINICAL IMPORTANCE: Quantitative PCR detection of B. bronchiseptica in BALF appears to be a useful diagnostic tool. Cytologic examination of BALF or BBF, when positive, allows a rapid and reliable diagnosis.