Effect of dasatinib in a xenograft mouse model of canine histiocytic sarcoma and in vitro expression status of its potential target EPHA2.

Canine histiocytic sarcoma (HS) is an aggressive and highly metastatic tumor. Previously, the kinase inhibitor dasatinib was shown to have potent growth inhibitory activity against HS cells in vitro, possibly via targeting the EPHA2 receptor. Here, the in vivo effect of dasatinib in HS cells was investigated using a xenograft mouse model. Moreover, the expression status of EPHA2 was examined in six HS cell lines, ranging from insensitive to highly sensitive to dasatinib. In the HS xenograft mouse model, dasatinib significantly suppressed tumor growth, as illustrated by a decrease in mitotic and Ki67 indices and an increase in apoptotic index in tumor tissues. On Western blot analysis, EPHA2 was only weakly detected in all HS cell lines, regardless of sensitivity to dasatinib. Dasatinib likely results in the inhibition of xenograft tumor growth via a mechanism other than targeting EPHA2. The findings of this study suggest that dasatinib is a targeted therapy drug worthy of further exploration for the treatment of canine HS.

Selective growth inhibition by suppression of F1Fo ATPase in canine malignant melanoma cell lines.

Canine malignant melanoma (CMM) is a highly aggressive and fatal neoplasm. To identify potential therapeutic compounds and/or targets, 320 compounds were screened for their growth inhibitory activity in a CMM line (CMM-1) using a chemical library known to target specific signaling pathways/cell growth-related molecules. Among the compounds screened, the F1Fo ATPase inhibitor oligomycin showed potent growth inhibitory effects in CMM-1 cells, while exhibiting less toxic effects in a non-neoplastic control cell line (MDCK cells). The growth inhibitory effect of oligomycin A was then examined using six CMM lines and MDCK cells. Three CMM lines were highly sensitive to oligomycin A, with around 3000-20 000 times lower IC50 compared with oligomycin A-resistant CMM lines and MDCK cells. Oligomycin A-sensitive CMM-1 cells exhibited much greater oligomycin A-induced decreases in cellular ATP compared to oligomycin A-resistant cell lines. Although the oligomycins are clinically unsuitable because of its in vivo toxicity, these findings implicate the potential of F1Fo ATPase as a therapeutic target in a subset of CMM.

Anticancer effects of gemcitabine are enhanced by co-administered iRGD peptide in murine pancreatic cancer models that overexpressed neuropilin-1.

BACKGROUND: Impaired drug transport is an important factor that reduces the efficacy of anticancer agents against pancreatic cancer. Here, we report a novel combination chemotherapy using gemcitabine (GEM) and internalised-RGD (iRGD) peptide, which enhances tumour-specific drug penetration by binding neuropilin-1 (NRP1) receptor. METHODS: A total of five pancreatic cancer murine models (two cell line-based xenografts (CXs) and three tumour grafts (TGs)) were treated with either GEM (100 mg kg(-1), q3d × 4) alone or GEM plus iRGD peptide (8 µmol kg(-1)). Evaluation of NRP1 expression in xenografts and 48 clinical cancer specimens was performed by immunohistochemistry (IHC). RESULTS: We identified a subset of pancreatic cancer models that showed NRP1 overexpression sensitive to iRGD co-administration. Treatment with GEM plus iRGD peptide resulted in a significant tumour reduction compared with GEM monotherapy in CXs, but not remarkable in TGs. Potential targets of iRGD were characterised as cases showing NRP1 overexpression (IHC-2+/3+), and these accounted for 45.8% of the clinical specimens. CONCLUSIONS: Internalised RGD peptide enhances the effects of co-administered drugs in pancreatic cancer models, its efficacy is however only appreciable in those employing cell lines. Therefore, the clinical application needs to be given careful consideration.

ACR Appropriateness Criteria® Orbital Imaging and Vision Loss-Child.

Orbital disorders in children consist of varied pathologies affecting the orbits, orbital contents, visual pathway, and innervation of the extraocular or intraocular muscles. The underlying etiology of these disorders may be traumatic or nontraumatic. Presumed location of the lesion along with the additional findings, such as eye pain, swelling, exophthalmos/enophthalmos, erythema, conjunctival vascular dilatation, intraocular pressure, etc, help in determining if imaging is needed, modality of choice, and extent of coverage (orbits and/or head). Occasionally, clinical signs and symptoms may be nonspecific, and, in these cases, diagnostic imaging studies play a key role in depicting the nature and extent of the injury or disease. In this document, various clinical scenarios are discussed by which a child may present with an orbital or vision abnormality. Imaging studies that might be most appropriate (based on the best available evidence or expert consensus) in these clinical scenarios are also discussed. The American College of Radiology Appropriateness Criteria are evidence-based guidelines for specific clinical conditions that are reviewed annually by a multidisciplinary expert panel. The guideline development and revision process support the systematic analysis of the medical literature from peer reviewed journals. Established methodology principles such as Grading of Recommendations Assessment, Development, and Evaluation or GRADE are adapted to evaluate the evidence. The RAND/UCLA Appropriateness Method User Manual provides the methodology to determine the appropriateness of imaging and treatment procedures for specific clinical scenarios. In those instances where peer reviewed literature is lacking or equivocal, experts may be the primary evidentiary source available to formulate a recommendation.

Mutations in a novel cochlear gene cause DFNA9, a human nonsyndromic deafness with vestibular dysfunction.

DFNA9 is an autosomal dominant, nonsyndromic, progressive sensorineural hearing loss with vestibular pathology. Here we report three missense mutations in human COCH (previously described as Coch5b2), a novel cochlear gene, in three unrelated kindreds with DFNA9. All three residues mutated in DFNA9 are conserved in mouse and chicken Coch, and are found in a region containing four conserved cysteines with homology to a domain in factor C, a lipopolysaccharide-binding coagulation factor in Limulus polyphemus. COCH message, found at high levels in human cochlear and vestibular organs, occurs in the chicken inner ear in the regions of the auditory and vestibular nerve fibres, the neural and abneural limbs adjacent to the cochlear sensory epithelium and the stroma of the crista ampullaris of the vestibular labyrinth. These areas correspond to human inner ear structures which show histopathological findings of acidophilic ground substance in DFNA9 patients.

Outcomes in Pediatric Thyroidectomy: Results From a Multinational, Multi-institutional Database.

OBJECTIVE: Traditionally, data regarding thyroidectomy were extracted from billing databases, but information may be missed. In this study, a multi-institutional pediatric thyroidectomy database was used to evaluate recurrent laryngeal nerve (RLN) injury and hypoparathyroidism. STUDY DESIGN: Retrospective multi-institutional cohort study. SETTING: Tertiary care pediatric hospital systems throughout North America. METHODS: Data were individually collected for thyroidectomies, then entered into a centralized database and analyzed using univariate and multivariable regression models. RESULTS: In total, 1025 thyroidectomies from 10 institutions were included. Average age was 13.9 years, and 77.8% were female. Average hospital stay was 1.9 nights and 13.5% of patients spent at least 1 night in the pediatric intensive care unit. The most frequent pathology was papillary thyroid carcinoma (42%), followed by Graves' disease (20.1%) and follicular adenoma (18.2%). Overall, 1.1% of patients experienced RLN injury (0.8% permanent), and 7.2% experienced hypoparathyroidism (3.3% permanent). Lower institutional volume (odds ratio [OR], 3.57; 95% CI, 1.72-7.14) and concurrent hypoparathyroidism (OR, 3.51; 95% CI, 1.64-7.53) correlated with RLN injury on multivariable analysis. Graves' disease (OR, 2.27; 95% CI, 1.35-3.80), Hashimoto's thyroiditis (OR, 4.67; 95% CI, 2.39-9.09), central neck dissection (OR, 3.60; 95% CI, 2.36-5.49), and total vs partial thyroidectomy (OR, 7.14; 95% CI, 4.55-11.11) correlated with hypoparathyroidism. CONCLUSION: These data present thyroidectomy information and complications pertinent to surgeons, along with preoperative risk factor assessment. Multivariable analysis showed institutional volume and hypoparathyroidism associated with RLN injury, while hypoparathyroidism associated with surgical indication, central neck dissection, and extent of surgery. Low complication rates support the safety of thyroidectomy in pediatric tertiary care centers.

Decrease of blood dendritic cells and increase of tissue-infiltrating dendritic cells are involved in the induction of Sjögren's syndrome but not in the maintenance.

We have demonstrated previously that, in primary Sjögren's syndrome (SS), immature myeloid dendritic cells (DCs) are decreased in blood and mature myeloid DCs are accumulated in salivary glands, suggesting recruitment of the myeloid DCs from blood to salivary glands. To verify whether this finding is universal in patients of not only primary SS but also secondary SS, in this study we analysed the blood DCs of secondary SS patients. We examined 24 secondary SS and 29 primary SS patients. A direct correlation between the decreased number of myeloid DCs and the duration of Sicca syndrome in primary and secondary SS was observed; namely, the reduction of myeloid DCs in blood was restored spontaneously with duration time of Sicca syndrome. We also examined the immunohistochemical staining of salivary glands of SS patients with monoclonal antibodies against fascin, CD11c and human leucocyte antigen DR (HLA-DR). Fascin(+) or CD11c(+)/HLA-DR(+) mononuclear cells were present in the salivary glands of secondary SS patients, as in primary SS. However, fascin(+) mononuclear cells were barely detected in the salivary glands of a chronic phase of SS patients. We also found a negative correlation between the frequency of blood myeloid DCs and salivary gland-infiltrating DCs in secondary SS patients, as well as primary SS. Our results suggest that the reduction of blood myeloid DCs and preferential trafficking of myeloid DCs into salivary glands is a common event in the early stage of SS. Myeloid DCs may play essential roles in the pathogenesis of Sicca syndrome of SS by initiating T helper cell immune responses.

Surgical management of obstructive sleep apnea in infants and young toddlers.

OBJECTIVE: Review surgical management of obstructive sleep apnea (OSA) in infants and young toddlers compared with a medically treated group. STUDY DESIGN: Case series with chart review of children younger than 24 months treated at a tertiary pediatric hospital between 2000 and 2005. SUBJECTS AND METHODS: Surgical treatment included adenotonsillectomy, adenoidectomy, and tonsillectomy. Polysomnography results, comorbidities, and major complications were recorded. The change in apnea-hypopnea index (AHI) before and after treatment was analyzed. Logistic regression analysis reviewed effects of comorbidities and OSA severity on complications. RESULTS: A total of 73 children met inclusion criteria. The surgical treatment group (AHI) improved posttreatment: mean AHI change was 9.6 (95% CI, 5.8-13.4). The medical treatment group did not improve posttreatment: mean AHI change was -3.0 (95% CI, -15.1 to 9.1). The difference in AHI change between surgical and medical groups was 12.56 (95% CI, 2.7-22.4). An independent t test found this difference to be statistically significant (P = 0.01). Eleven (18%) patients suffered significant postoperative surgical complications; 55 surgical patients and 8 medical patients had comorbidities. There were no long-term morbidities or mortalities. CONCLUSIONS: AHI in the surgically treated group significantly improved. The complication rate for a tertiary pediatric hospital population that included patients with multiple comorbidities was acceptable.

Increased time between diagnosis and surgery in slipped capital femoral epiphysis results in increased radiographic deformity.

PURPOSE: Previous work has examined the impact of delay of diagnosis in slipped capital femoral epiphysis (SCFE) but not the impact of delay in treatment after radiographic diagnosis. Due to requirements for long distance transportation from less developed regions for many of our patients, our hospital was able to study variation in time between diagnosis and surgery for SCFE, as related to slip severity. METHODS: This is a retrospective review of patients treated for SCFE between 2005 and 2014 at a tertiary care paediatric hospital. Demographics, time between diagnosis and surgery, radiographic deformity (Southwick angle), postoperative complications and need for further surgery were variables of interest. Statistical analysis included Pearson and Spearman rank correlations and chi-squared tests. RESULTS: The study sample included 147 hips (119 patients). Mean time between radiographic diagnosis and surgery was 20.9 days (sd 46, 0 to 321). The mean Southwick angle (SA) at the time of surgery was 31.9˚ (sd 19.6˚, 1° to 83˚). There was a significant relationship between increased delay and increased SA (0.34, p < 0.001). Increased SA was correlated with need for future significant surgery (0.27, p < 0.01).Patients from less-developed regions, with barriers to timely care, had moderate and severe deformity (SA) (p < 0.01), and required significant further surgery more often than SCFE patients from the local population (p < 0.01). CONCLUSION: The unique referral environment of our hospital provided an opportunity to examine traditional recommendations for treating SCFE promptly after radiographic diagnosis. Delay in treatment is correlated with increased radiographic deformity. LEVEL OF EVIDENCE: III.

Canine squamous cell carcinoma cell lines with high expression of survivin are sensitive to survivin inhibitor YM155.

Treatment of unresectable canine squamous cell carcinoma (SCC) remains challenging and new therapeutic strategies are needed. Survivin is a member of the inhibitor of apoptosis protein family and its inhibitor, YM155, is a potential anti-tumour agent. In the present study, 10 canine tumour cell lines (representing eight different tumour types) were screened for sensitivity to YM155; the drug potently inhibited the growth of the HAPPY SCC cell line. The growth inhibitory properties of YM155 were then examined in more detail using a panel of seven SCC cell lines. YM155 inhibited the growth of the cell lines HAPPY and SQ4; in contrast to the other lines in the panel, these two cell lines had high levels of expression of survivin. In HAPPY cells, YM155 inhibited expression of the survivin gene at the transcriptional level. In contrast, YM155 down-regulated survivin at the post-transcriptional level in SQ4 cells. YM155 suppressed cell growth in HAPPY cells, mostly via induction of apoptosis, but this was not the case in SQ4 cells. Two canine SCC cell lines with high cellular expression of survivin were sensitive to YM155. The possible underlying mechanisms of the cytotoxic effect of YM155 in these cell lines were different. One cell line had down-regulation of survivin mRNA and protein expression, associated with induction of apoptotic cell death. The other cell line had post-transcriptional down-regulation of survivin expression and subsequent induction of non-apoptotic cell death. Targeting survivin with YM155 is a potential approach for the treatment of canine SCCs with high expression of survivin.

Behavioral inhibition and clinical outcomes in children with cochlear implants.

OBJECTIVES/HYPOTHESIS: Individual speech and language outcomes of deaf children with cochlear implants (CIs) are quite varied. Individual differences in underlying cognitive functions may explain some of this variance. The current study investigated whether behavioral inhibition skills of deaf children were related to performance on a range of audiologic outcome measures. DESIGN: Retrospective analysis of longitudinal data collected from prelingually and profoundly deaf children who used CIs. METHODS: Behavioral inhibition skills were measured using a visual response delay task that did not require hearing. Speech and language measures were obtained from behavioral tests administered at 1-year intervals of CI use. RESULTS: Female subjects showed higher response delay scores than males. Performance increased with length of CI use. Younger children showed greater improvement in performance as a function of device use than older children. No other subject variable had a significant effect on response delay score. A series of multiple regression analyses revealed several significant relations between delay task performance and open set word recognition, vocabulary, receptive language, and expressive language scores. CONCLUSIONS: The present results suggest that CI experience affects visual information processing skills of prelingually deaf children. Furthermore, the observed pattern of relations suggests that speech and language processing skills are closely related to the development of response delay skills in prelingually deaf children with CIs. These relations may reflect underlying verbal encoding skills, subvocal rehearsal skills, and verbally mediated self-regulatory skills. Clinically, visual response delay tasks may be useful in assessing behavioral and cognitive development in deaf children after implantation.

Bilateral congenital vocal cord paralysis: a 16-year institutional review.

OBJECTIVE: To review the management and outcome of bilateral congenital true vocal cord paralysis in 22 patients treated over a 16-year period and to review the role of tracheostomy in these patients. DESIGN: Retrospective chart review. SETTING: Pediatric tertiary hospital. PATIENTS: Twenty-two pediatric patients diagnosed with bilateral congenital true vocal cord paralysis. INTERVENTIONS: Flexible or rigid diagnostic evaluation, tracheostomy, and vocal cord lateralization procedures. MAIN OUTCOMES MEASURES: Vocal cord recovery and decannulation. RESULTS: With a mean follow up of 50 months, 15 of 22 patients (68%) with bilateral vocal cord paralysis required tracheostomy for airway securement. Of the 15 tracheotomized patients, 10 were successfully decannulated (8 had spontaneous recovery, whereas 2 required lateralization procedures). Eleven of these patients with tracheostomy had comorbid factors, including neurologic abnormalities (midbrain/brainstem dysgenesis, Arnold-Chiari malformation, global hypotonia, and developmental delay). Of the 7 patients not requiring tracheostomy, 6 recovered vocal cord function (86%). CONCLUSION: In our series of 22 patients with bilateral vocal cord paralysis, 14 had spontaneous recovery of function. Patients managed with tracheostomy were noted to have a high incidence of comorbid factors. In this series, recovery rates were found to be higher in nontracheostomized patients than in tracheostomized patients. Patients can be carefully selected for observation versus tracheostomy at the time of diagnosis based on underlying medical conditions.

Infantile muscle glycogen storage disease: phosphoglucomutase deficiency with decreased muscle and serum carnitine levels.

We report a 5-month-old boy with recurrent vomiting, lethargy, and poor weight gain. He had profound metabolic acidosis and nonketotic dicarboxylic aciduria. The serum and muscle carnitine levels were significantly low (60% and 10% of the control means, respectively), suggesting that the patient had a systemic carnitine deficiency syndrome. The patient showed apparent clinical improvement on oral carnitine administration. A quadriceps muscle biopsy revealed a slight increase in intrafiber lipid droplets and mild accumulation of glycogen in the subsarcolemmal portion. An anaerobic glycolysis in vitro study showed a block after glucose-1-phosphate and before glucose-6-phosphate. Direct measurement of individual glycolytic enzymes in muscle of the patient demonstrated a marked decrease in phosphoglucomutase (PGM) activity (13% of the control mean). The specific defect of PGM activity in this patient suggests that the block in the anaerobic glycolytic pathway is the primary abnormality. PGM deficiency can be added as a newly recognized cause of secondary systemic carnitine deficiency syndromes.

Tolerance of NK and LAK activity for HLA class I-deficient targets in a TAP1-deficient patient (bare lymphocyte syndrome type I).

NK cells recognize target cells that lack HLA class I molecules and lyse them, according to the 'missing self' hypothesis. It was previously reported that a TAP2-deficient patient with an HLA class I-deficiency, had a normal number of NK cells but that the lymphocytes of this patient had lost their NK activity against K562 cells. In this study, we investigated the HLA class I-recognizing NK receptor expressions and the NK and LAK activities of the lymphocytes of a TAP1-deficient patient. The patient had a normal number of NK cells. Although the lymphocytes showed LAK activity against class I expressing targets following IL-2, IL-12 and IL-15 stimulation for 3 days, neither NK nor LAK activity against targets lacking class I molecules was induced. The NK cells of the patient expressed class I-recognizing NK receptors, although the percentages of such cells were low. However, no differences were observed in the expression levels of inhibitory and activating NK receptors between lymphocytes of the patient and those of healthy controls, suggesting that the modulation of the NK receptor expression is not primarily responsible for this tolerance. These results also suggest that the lymphocytes of the patient are defective in the recognition of class I-deficient target cells in order to promote the induction of self tolerance.

Cultural identification and attempted suicide in Native Hawaiian adolescents.

OBJECTIVES: To determine rates of lifetime suicide attempts in a community sample of Native Hawaiian adolescents and determine the contribution of Hawaiian cultural affiliation, socioeconomic status, and psychiatric symptoms as risk factors for suicide. METHOD: High school students were surveyed in the state of Hawaii for lifetime suicide attempts, Hawaiian cultural affiliation, socioeconomic status, and symptoms of depression, substance abuse, aggression, and anxiety. Multiple logistic regressions were used on 3,094 subjects to develop prediction models for lifetime suicide attempts. RESULTS: Native Hawaiian adolescents had significantly higher rates of suicide attempts (12.9%) than other adolescents in Hawaii (9.6%). Hawaiian cultural affiliation rather than ethnicity was uniquely predictive of suicide attempts. Logistic regression indicated that depression, substance abuse, grade level, Hawaiian cultural affiliation, and main wage earner's education best predicted suicide attempts in Native Hawaiian adolescents, while depression, substance abuse, and aggression predicted suicide attempts in non-Hawaiians. CONCLUSIONS: Native Hawaiian adolescents have higher rates of attempted suicide than non-Hawaiian adolescents. Strong Hawaiian cultural affiliation rather than ethnicity is a risk factor for attempted suicide.

Prediction of major depression and dysthymia from CES-D scores among ethnic minority adolescents.

OBJECTIVE: The Native Hawaiian Mental Health Research Development Program is an epidemiological longitudinal study of adolescents residing in Hawaii. This article examines the utility of the Center for Epidemiologic Studies-Depression Scale (CES-D) for predicting DSM-III-R diagnoses of major depression (MD) and dysthymic disorder (DD) and investigates whether prediction differs by gender and ethnicity. METHOD: Diagnostic Interview Schedule for Children interviews were conducted with 556 adolescents randomly selected from among more than 7,000 students who had completed the CES-D. RESULTS: Six-month prevalence rates were as follows: MD = 8.5%, DD = 4.7%, either (MDDD) = 9.9%. Prevalence rates were significantly higher among females, but after CES-D scores were accounted for, gender no longer predicted depression in most analyses. When a cutoff score of 16 was used, classification accuracy was lower for Native Hawaiians than non-Hawaiians. However, after group differences in gender and grade level were accounted for, the predictive validity of the CES-D did not differ by ethnicity. CES-D factor 1 scores identified MD, DD, and MDDD about as well as the total score or all three factors together. CONCLUSIONS: These results support the validity of the CES-D for screening for depression among adolescents of Native Hawaiian and other minority backgrounds.

Effect of chronic administration of acetaldehyde by inhalation on (Na+ + K+)-activated adenosine triphosphatase activity of rat brain membranes.

The effect of chronic acetaldehyde inhalation on (Na+ + K+)-ATPase (EC 3.6.1.3) activities of subcellular fractions of the rat cerebral cortex was studied. Chronic administration of acetaldehyde by inhalation for 4-21 weeks caused significant increases in the enzyme activities of both the synaptosomal plasma membrane (SPM) fraction and the microsomal (MC) fraction. This indicates the change in neural membrane functions of the brain after acetaldehyde treatment. Mg2+-ATPase activities of both subcellular fractions remained unchanged after acetaldehyde treatment.

Subcellular aldehyde dehydrogenase activity and acetaldehyde oxidation by isolated intact mitochondria of rat brain and liver after acetaldehyde treatment.

The effect of treatment of rats with acetaldehyde on the subcellular NAD+-aldehyde dehydrogenase (EC 1.2.1.3, ALDH) activities and acetaldehyde oxidation by isolated intact mitochondria of the liver and the brain was studied. Inhalation of acetaldehyde caused a significant decrease in the liver mitochondrial low Km-ALDH activity, while brain mitochondrial ALDH activity remained unchanged. Acetaldehyde oxidation by isolated intact liver mitochondria decreased significantly but that by brain mitochondria remained unchanged after acetaldehyde inhalation. These findings raise the possibility that the brain enzyme may be exposed to lower concentration of acetaldehyde than the liver enzyme.

Circadian rhythms in the activities of brain and liver aldehyde dehydrogenase isozymes in mice.

Circadian variations in the activities of aldehyde dehydrogenase (ALDH) isozymes in the subcellular fractions of the brain and liver were investigated in male and female mice of C57BL/6J strain. The rhythms in high Km-ALDH activities of brain and liver mitochondrial fractions which existed in ordinary light-dark cycle were not observed in animals maintained in the continuous darkness for two weeks. The rhythms in high Km-ALDH activities of hepatic soluble and microsomal fractions existed in both ordinary cycle and total darkness but the rhythmic phases were different. In the low Km-ALDH activity of hepatic mitochondrial fraction, the circadian rhythm was similar in two lighting conditions. There was sex difference in the existence of the circadian rhythm. It seems that the ALDH activity of mice is influenced by light-dark cycle and sex hormones.