Evaluation of ponatinib in vitro effect in three canine mast cell tumor cell lines expressing FGFR-1, PDGFR-α, and VEGFR-2.

Ponatinib is a broad-spectrum tyrosine kinase inhibitor that targets numerous receptor tyrosine kinases (RTKs), including but not limited to fibroblast growth factor receptor (FGFR)-1, platelet derived growth factor receptor (PDGFR)-α, and vascular endothelial growth factor receptor (VEGFR)-2. This study evaluated the expression of FGFR-1, PDGFR-α, and VEGFR-2 in three canine mast cell tumor (MCT) cell lines (CM-MC1, VI-MC1, CoMS) and the effects of ponatinib on these MCT cell lines. Quantitative RT-PCR confirmed the expression of FGFR-1, PDGFR-α, and VEGFR-2 in the three MCT cell lines. Ponatinib exhibited dose- and time-dependent cytotoxicity in MCT cell lines via MTT assay. The IC50 for 24, 48, and 72 h across the three cell lines ranged from 38.47 nM to 103.3 nM, which is clinically comparable to dose ranges established for humans. Significantly increased apoptosis in each cell line was seen between 12 and 18 h after treatment with IC50 of ponatinib via Annexin-V and Caspase-3/7 assays. These data suggest that ponatinib could be a possible therapeutic agent for canine MCTs. Further studies are needed to investigate the prognostic value of FGFR-1, PDGFR-α, and VEGFR-2 in canine MCTs.

c-Jun enhancement of androgen receptor transactivation is associated with prostate cancer cell proliferation.

Androgens and the androgen receptor (AR) are involved in the growth and progression of prostate cancer. Our previous studies suggest that the proto-oncoprotein c-Jun is an AR coactivator that stimulates AR transactivation by mediating receptor dimerization and subsequent DNA binding. To study the physiological relevance of this c-Jun activity on AR, we have generated stable LNCaP cell lines expressing different levels of c-Jun. These cell lines exhibit a direct correlation between endogenous c-Jun levels and AR transcriptional activity and expression of endogenous androgen-regulated genes. Disruption by antisense RNA of endogenous c-Jun expression in LNCaP cells strongly compromises the androgen-dependent proliferation of these cells. In contrast, expression of a c-Jun mutant, which is fully active in coactivation of AR but deficient in AP-1 transactivation, significantly enhances androgen-dependent proliferation. This finding indicates that the coactivation function of c-Jun is sufficient for regulating androgen-induced growth of LNCaP cells. c-Jun also enhances AR transactivtion in androgen-independent LNCaP cells, which closely mimic hormone-refractory prostate cancer cells in gene expression and growth behavior. Importantly, siRNA-mediated repression of endogenous c-Jun expression results in markedly reduced growth of these cells, strongly suggesting an important biological role for c-Jun in hormone-refractory prostate cancer.

Transformation-induced alterations in the regulation of tyrosine aminotransferase expression in fetal rat hepatocytes: changes in hormone inducibility and the DNase-hypersensitive site.

Regulation of expression of tyrosine aminotransferase (TAT) is examined in two cell lines (FRL) obtained by chemical transformation of cultured fetal hepatocytes derived from 19-day rat fetuses (FL19). Steroid induction of TAT is unaffected by transformation, while the response to cyclic AMP is attenuated. Consequently a synergistic response elicited by the simultaneous exposure of normal fetal hepatocytes to the inducers is almost abolished in FRL cells. FL19 and FRL are similar with respect to the negative effect of insulin on steroid induction, which is a response restricted to prenatal liver. Detailed examination of chromatin reveals that the attenuated effect of cyclic AMP is consistent with the lack of the DNase I-hypersensitive site located at about the cyclic AMP response element of the TAT promoter. From the studies, we conclude that transformation results in the modification of some aspects of TAT regulation, while others have been retained, and reflects the fetal pattern which is observed in normal embryonic hepatocytes.

Prolonged bromide intoxication resulting from a gastric bezoar.

Bromide intoxication is now a relatively uncommon disease. We describe a critically ill patient who had an elevated chloride level and neurologic abnormalities. Bromide levels were also elevated, and remained so for several days. After more than 12 days of abnormally high serum bromide and chloride levels, endoscopy was performed. This procedure disclosed a gastric bezoar containing identifiable pill fragments. The gastric washings from these fragments contained high levels of bromide. We believe that the prolonged course of bromide intoxication secondary to a gastric bezoar makes this case unique.

Immunohistochemical detection of p53 protein in mammary carcinoma: an important new independent indicator of prognosis?

In an immunohistochemical pilot study of 195 primary breast cancer patients with a 10-year median follow-up we found that patients with carcinomas who express p53 protein in the majority of their tumor cells (19% of the cases) have a considerably worse prognosis than those who do not. The effect of the presence of the protein is seen on disease-free interval (chi-square, 11.69; P < .001), overall survival (chi-square, 19.68; P < .001), and survival after relapse (chi-square, 4.93; P < .02), and is seen in node-negative (chi-square, 6.99; P < .009) and node-positive (chi-square, 13.05; P < .001) patients. Furthermore, the effect is most apparent in patients with infiltrating lobular and grade II infiltrating ductal carcinomas (chi-square, 27.97; P < .001) that have a rather heterogeneous clinical behaviour and are difficult to subdivide on the basis of currently available markers. Cox multivariate analysis shows that p53 majority staining is second only to node status in significance of effect on overall survival.

Early methylprednisolone treatment for septic syndrome and the adult respiratory distress syndrome.

From November 1, 1982 through December 31, 1985, there were 19 centers and 382 patients that evaluated the effect of methylprednisolone sodium succinate (MPSS) on the septic syndrome. Seventeen of these centers enrolled 304 patients in a prospective, randomized, double-blind, placebo-controlled study to determine if early treatment with MPSS would decrease the incidence of severity of the adult respiratory distress syndrome (ARDS) in patients at risk of ARDS from sepsis. To ensure early institution of the MPSS or placebo therapy (PLA), patients with the presumptive diagnosis of sepsis were identified. That diagnosis was based on the presence of fever or hypothermia (temperature greater than 38.3 degrees C or less than 35.5 degrees C, rectal), tachypnea (greater than 20 bpm), tachycardia (greater than 90 bpm) and the presence of one of the following indices of organ dysfunction: a change in mental status, hypoxemia, elevated lactate levels or oliguria. The treatment, either MPSS 30 mg/kg or PLA, was given in four 20-minute infusions six hours apart and was initiated within two hours of the presumptive diagnosis of sepsis. The development and reversal of the adult respiratory distress syndrome (ARDS) was followed and resulted in data on 304 of the 382 randomized patients. A trend toward increased incidence of ARDS was seen in the MPSS group 50/152 (32 percent) compared to the placebo group 38/152(25 percent) p = 0.10. Significantly fewer MPSS patients reversed their ARDS 15/50 (31 percent) compared to placebo 23/38 (61 percent) p = 0.005. The 14-day mortality in patients with ARDS treated with MPSS was 26/50 (52 percent) compared to placebo 8/22 (22 percent) p = 0.004. We conclude that early treatment of septic syndrome with MPSS does not prevent the development of ARDS. Additionally, MPSS treatment impedes the reversal of ARDS and increases the mortality rate in patients with ARDS.

Selective left endobronchial suctioning in the intubated patient.

Suctioning of secretions from the left endobronchial tree is frequently necessary but often difficult in intubated patients. We examined the effectiveness of a catheter designed expressly for this purpose. Special curved tip (Bronchitrac-L) suction catheters were fitted with thin, radiopaque tubing to facilitate x-ray visualization. Eight-one attempts at left endobronchial placement were made on 74 stable adult intensive care unit patients. The suction catheter was inserted into the oral endotracheal tube or tracheostomy tube just prior to an x-ray filming of the chest. In 15 of 66 patients, the tip of the oral endotracheal tube was too distal (less than 2 cm above the carina) to allow proper functioning of the catheter. Patients with a properly positioned oral endotracheal tube were analyzed separately and showed 56 percent of the catheters went to the left bronchus. When the head was turned to the left prior to placement, successful left bronchus placement occurred in 65 percent. When the catheter was placed through a tracheostomy tube, 100 percent went into the left bronchus (n = 15). There were no catheter-induced complications in this study. The curved tip catheter is an effective means of suctioning the left bronchial tree in patients with tracheostomy tubes. Its reliability in patients with oral endotracheal tubes is reduced but more effective than current methods.

Benign lymph node inclusions mimicking metastatic carcinoma.

AIMS: To draw attention to non-neoplastic inclusions in axillary lymph nodes removed from women with primary breast cancer which may be mistaken for metastases. METHODS: Five examples of non-malignant inclusions were detected in axillary lymph nodes removed from women with mammary carcinoma. Immunohistochemical staining for CAM 5.2 and S100 markers, as well as morphological assessment were performed. RESULTS: Three of the five cases comprised naevus cells and two heterotopic epithelial elements. One of each was initially mistaken for metastatic carcinoma. Naevus cells in the capsule of axillary nodes are often arranged as lines of small, round cells and may readily be misinterpreted as metastatic lobular carcinoma. Heterotopic epithelial elements, in the form of tubules, can easily be mistaken for metastasis from an infiltrating ductal carcinoma. CONCLUSIONS: Awareness of the occasional occurrence of non-neoplastic nodal inclusions will help avoid misdiagnosis. If immunohistochemical markers for epithelial cells are used to screen lymph nodes for metastasis, such lesions could be detected more frequently. It is important to combine immunohistochemistry with morphological assessment and the use of a panel of antibodies in histological diagnosis.

Accidental mediastinal entry via left internal jugular vein cannulation.

Two cases are reported in which mediastinal penetration by a pulmonary artery catheter and a temporary venous pacemaker wire occurred following cannulation of the left internal jugular vein and placement of an indwelling venous introducer sheath. The anatomy of the left internal jugular vein and possible mechanisms of accidental mediastinal penetration with this approach are discussed. Extreme caution must be exercised when using the left internal jugular venous access route and an indwelling venous introducer sheath.

Physiology of respiration.

Patients with significant respiratory disease are evaluated daily by emergency physicians. Understanding the principles of respiratory physiology and pathophysiology provides the basis for the diagnosis and management of patients with respiratory disease.

A unique academic approach to critical care-emergency medicine training.

The four-year program for training physicians in critical care-emergency medicine at Sacramento Medical Center, University of California, Davis School of Medicine is described. Its purpose is to train the physician in resuscitation, emergency care for life-threatening conditions, and intensive care, so that the graduate can staff community hospital emergency departments and intensive care units, be director of an emergency department or intensive care unit, or pursue an academic career in emergency medicine.

Mobile triage team in a community disaster plan.

Experience has shown poor predisaster planning, inadequate communication and the absence of an on-scene commander to be common and recurring problems during disaster rescue efforts. A mobile onscene triage team (MOTT) operating in Sacramento has demonstrated the following advantages: immediate access; mobility; coordinated evacuation, treatment, and disposition of mass casualty victims; control of facility overload, and appropriate initial disposition to definitive care facilities. The advantages realized with this approach arise from greater community awareness and participation in a coordinated plan for medical care in disasters.

Protein C levels as a prognostic indicator of outcome in sepsis and related diseases.

OBJECTIVE: To consider the appropriateness of protein C levels as a prognostic indicator for sepsis and related diseases. DATA SOURCES/STUDY SELECTION: Published research and review articles related to protein C deficiency in patients with sepsis and related diseases. DATA EXTRACTION AND SYNTHESIS: All applicable data were extracted, and relevant literature was cited to support factual statements in the text. The protein C pathway represents one of the major regulatory systems of hemostasis, exhibiting antithrombotic, profibrinolytic, and anti-inflammatory properties. Numerous studies have shown that acquired protein C deficiency is prevalent in the majority of septic patients (>85%) and is associated with increased morbidity and mortality in patients with severe sepsis and septic shock. This deficiency in protein C is not simply a transient marker for sepsis, but parallels the progress of the disease. In addition, protein C deficiency occurs in the presence of a wide range of pathogens and develops early in the disease process. CONCLUSIONS: A review of the relevant literature suggests that protein C levels may serve as a useful prognostic indicator of outcome in sepsis and related diseases.

Immunotherapy of sepsis syndrome: a comparison of the available treatments.

OBJECTIVE: To evaluate the current range of immunotherapy for sepsis syndrome. DATA: Three major, recent anti-endotoxin and two human polyclonal, immunoglobulin clinical trials are reviewed. COMPARISONS: Comparisons are made in their trial design, definitions, patient selection and exclusion criteria and patient demographics. The results in specific subsets are compared and reasons for the differences in efficacy, discussed. RESULTS: J5 anti-sera, E-5, and HA-1A are all effective in reducing the mortality in patients with gram-negative infections. They also result in enhanced resolution of organ failure. Similarities in the efficacies of J5 anti-sera and HA-1A for bacteremic patients with and without shock are noted. High-dose polyclonal human immunoglobulins have been shown to be of benefit in both gram-positive and gram-negative infections in two small clinical trials and deserve further evaluation. CONCLUSION: Anti-endotoxin monoclonal antibodies benefit patients with gram-negative sepsis. They are proof of a concept, but are costly and do not show an overall intention-to-treat effect. There is an urgent need to accurately identify, clinically or by a cytokine profile, those patients who will specifically benefit.