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Abstract DNA replication is a critical step in cell proliferation. Overexpression of MCM2-7 genes correlated with poor prognosis in breast cancer patients. However, the roles of Cdc6 and Cdt1, which work with MCMs to regulate DNA replication, in breast cancers are largely unknown. In the present study, we have shown that the expression levels of Cdc6 and Cdt1 were both significantly correlated with an increasing number of MCM2-7 genes overexpression. Both Cdc6 and Cdt1, when expressed in a high level, alone or in combination, were significantly associated with poorer survival in the breast cancer patient cohort (n = 1441). In line with this finding, the expression of Cdc6 and Cdt1 was upregulated in breast cancer cells compared to normal breast epithelial cells. Expression of Cdc6 and Cdt1 was significantly higher in ER negative breast cancer, and was suppressed when ER signalling was inhibited either by tamoxifen in vitro or letrozole in human subjects. Importantly, breast cancer patients who responded to letrozole expressed significantly lower Cdc6 than those patients who did not respond. Our results suggest that Cdc6 is a potential prognostic marker and therapeutic target in breast cancer patients.

Bladder cancer is one of the leading causes of cancer-related death in men, however, there was only limited effective treatment for invasive bladder cancer. DAPK1 has been shown to play important role in apoptosis and autophagy to suppress cancer progression. Previous results have shown that DAPK1 promoter was hypermethylated in the majority of bladder cancer specimens, however, the prognostic significance of DAPK1 in bladder cancer has yet to be demonstrated. In the present study, we found that DAPK1 expression was negatively associated with tumor stage and a low level expression of DAPK1 in bladder cancer specimens were associated with shorter survival in bladder cancer patients in 3 independent bladder cancer datasets (n = 462). Further investigation showed that FGFR3 knockdown resulted in downregulation of DAPK1 in bladder cancer cell line, suggesting that FGFR3 may be an upstream factor of DAPK1. Further analysis of the 3 independent bladder cancer datasets have identified ACOX1, UPK2, TRAK1, PLEKHG6 and MT1X genes had their expression significantly correlated with that of DAPK1. Knockdown of DAPK1 in bladder cancer T24 cells resulted in downregulation of ACOX1, UPK2 and TRAK1. Interestingly, TRAK1, by itself, was a favorable prognostic marker in the 3 independent bladder cancer datasets. Importantly, by using connectivity mapping with DAPK1-associated gene signature, we found that vemurafenib and trametinib could possibly reverse DAPK1-associated gene signature, suggesting that inhibition of Raf/MEK pathway may be a potential therapeutic approach for bladder cancer. Indeed, treatment of vemurafenib in T24 bladder cancer cells resulted in upregulation of DAPK1 confirming our connectivity mapping, while knockdown of DAPK1 resulted in reduced sensitivity towards inhibition of Braf signaling by vemurafenib. Together, our results suggest that DAPK1 is an important prognostic marker and therapeutic target for bladder cancer and have identified possible therapeutic agents for future testing in bladder cancer models with low DAPK1 expression.

IntroductionSpirometry devices, which are components of many anaesthesia machines, are commonly used to assess lung mechanics during anaesthesia. Spirometry calibration usually adheres to manufacturer recommendations without established guidelines. Although more accurate and less variable than inbuilt spirometry in certain General Electric anaesthesia ventilators, near-patient spirometry lacks adequate evaluation.MethodsWe assessed near-patient spirometers’ performance using Pedi-lite and D-lite flow sensors. Certified 1 L calibration syringes were used on 67 monitors located in 14 veterinary hospitals. Three consecutive inspired and expired volume values displayed by the monitors for each volume of the calibration syringe were recorded. Volumes studied were 50, 100, 150, 250, 300 mL for Pedi-lite and 150, 300, 450, 500, 750 mL for D-lite. Measured and targeted volumes were averaged, agreement error calculated. Accuracy was assessed plotting agreement errors against calibration volumes. A linear mixed-effects model was used to obtain linear regression between the error and the calibration volume. Mean, differential and proportional bias, limits of agreement, claimed accuracy and 10% clinical tolerance were calculated and displayed. Differences among monitors were evaluated using the Friedman rank sum test, differences between inspired and expired volumes using the Wilcoxon signed-rank.ResultsInter-monitor variability for inspired and expired volume readings using both sensors was high; intra-monitor variability was low. The error magnitude was independent of volumes evaluated. Using Pedi-lite, only a minority of measurements met manufacturer’s specification or a 10% clinical tolerance; both inspired and expired volumes were significantly underestimated. Using D-lite, superior performance was demonstrated for volumes between 300 and 750 mL (mean biases close to zero and the majority of measurements meeting manufacturer’s specifications and clinical tolerance). The difference between measured inspired and expired volumes with both sensors was significant.DiscussionThese results support caution when interpreting clinical measurements of lung volumes and mechanics in anaesthetised patients when using these sensors. This is particularly important in smaller patients where lung volumes are below 300 mL. Trends should be reliable.

IntroductionIn clinical practice, evaluating dynamic compliance of the respiratory system (Cdyn) could provide valuable insights into respiratory mechanics. Reference values of Cdyn based on body weight have been reported, but various factors may affect them and the evidence is scanty. This study aimed to establish a reference interval for Cdyn and identify associated variables.MethodsData were collected from 515 client-owned dogs requiring anesthesia, excluding those with lower airway disease. The dogs were anesthetized, the tracheas intubated, and lungs ventilated at clinicians' discretion across 11 centers in six countries, with no restrictions on anesthesia protocols or ventilation settings, except avoiding inspiratory pauses. Three Cdyn measurements from three consecutive breaths per dog were recorded using a standardized form, which also documented factors affecting Cdyn identified through literature and an online survey. Various spirometry technologies were used. The substantial variance in Cdyn measurements led to a comprehensive analysis using a multiple linear regression model. Multicollinearity (variables highly correlated with each other) was addressed by investigating, transforming, or excluding factors. Initial simple linear regression assessed each variable's individual effect on Cdyn, followed by a multiple linear regression model constructed via stepwise forward selection and backward elimination.ResultsThe best-fitting model identified a linear relationship between Cdyn and body mass when the following conditions were met: high BCS (Body Condition Score), orotracheal tubes 80%) exposure for more than 10 minutes before Cdyn measurement. In cases where these conditions were not met, additional factors needed to be incorporated into the model. Low (1/9, 2/9, 3/9) and medium (4/9, 5/9) BCS, an orotracheal tube of the predicted size or larger and longer inspiratory times were associated with increased Cdyn. The use of alternative spirometry sensors, including Ped-lite, or prolonged exposure to high FIO2 levels resulted in decreased Cdyn.Conclusion and clinical relevanceEstablishing a reference interval for Cdyn proved challenging. A single reference interval may be misleading or unhelpful in clinical practice. Nevertheless, this study offers valuable insights into the factors affecting Cdyn in healthy anesthetized dogs, which should be considered in clinical assessments.

BACKGROUND: Infection/reactivation of cytomegalovirus is a major cause of morbidity and mortality in immunocompromised transplant patients. It has already been observed in kidney and liver transplantation patients that cytomegalovirus disease is accompanied by significant increases in circulating CD8+CD38+ T lymphocytes. There are no reports that study CD8+CD38+ T lymphocytes to monitor/diagnose cytomegalovirus disease in hematopoietic stem cell transplantation patients. OBJECTIVE: The aim of this study was to evaluate some cellular activation markers on circulating mononuclear cells (CD38 and HLA-DR) in patients submitted to hematopoietic stem cell transplantation and to establish any correlation with cytomegalovirus disease as diagnosed by antigenemia. METHODS: Blood samples of 15 transplant patients were analyzed by flow cytometry using anti-CD3, anti-CD4, anti-CD8, anti-CD38, CD16, CD56 and anti-HLA-DR monoclonal antibodies and the results were evaluated in respect to cytomegalovirus antigenemia measured by indirect immunofluorescence. Minitab for Windows was used for statistical analysis and a p-value < 0.05 was considered significant. RESULTS: Patients with positive antigenemia did not show any significant increase in the percentages of cells expressing the CD38 or HLADR activation markers when compared to patients with negative antigenemia. On the contrary, all patients showed high percentages of these cells independent of the presence of cytomegalovirus disease. CONCLUSIONS: This study suggests that the investigation of these lymphocyte sub-populations in patients submitted to hematopoietic stem cell transplantation does not seem to contribute to the early identification of cytomegalovirus disease.

We report an increased incidence of high relapse risk features in 157 APL Brazilian patients. Out of 134 patients treated with ATRA and anthracyclines, only 91 (67.9%) achieved remission because 43 (32%) died during induction. The death rate during consolidation was 10.5%. Bleeding complications were the most frequent cause of failure (21.6%).

The purpose of this study was to compare cardiovascular and respiratory effects of two volumes of bupivacaine 0.25% (0.2 mL kg-1-treatment BUP02-and 0.4 mL kg-1 -treatment BUP04) administered epidurally at the lumbosacral intervertebral space in dogs anesthetized with isoflurane. This experimental prospective randomized crossover design trial used six mixed breed adult dogs, four neutered males and two spayed females. Each dog was anesthetized on three different occasions: the first for isoflurane minimum alveolar concentration (MAC) measurement, and the following two assigned treatments (BUP02 or BUP04). On the two treatment days, anesthesia was induced and maintained with isoflurane at 1.3 MAC during the experiments. Cardiovascular and respiratory measurements were recorded before (T0) and 5, 15, 30, 60 and 90 minutes after the epidural administration of bupivacaine. Comparisons between and within groups were performed by a mixed-model ANOVA and Friedman's test when appropriate followed by Bonferroni post-hoc test or Dunnet's test to compare time points within each treatment with T0 (p < 0.05). Mean arterial pressure decreased significantly from 15 to 90 minutes after the administration of BUP02 and from 5 to 60 minutes in BUP04, with lower values in BUP04 than in BUP02 lasting up to 30 minutes after bupivacaine administration. No significant changes in cardiac output and systemic vascular resistance were observed in either treatment. Hypoventilation was only detected in BUP04. Hemoglobin concentration and arterial oxygen content decreased after both treatment of bupivacaine with no significant decrease in oxygen delivery. Two dogs in BUP04 developed Horner's syndrome. The epidural administration of 0.4 mL.kg-1 of bupivacaine to dogs in sternal recumbency anesthetized with isoflurane 1.3 MAC caused more cardiovascular and respiratory depression than 0.2 mL.kg-1.