Establishment of canine mammary gland tumor cell lines harboring PI3K/Akt activation as a therapeutic target.

Canine mammary gland tumors (MGT) have a poor prognosis in intact female canines, posing a clinical challenge. This study aimed to establish novel canine mammary cancer cell lines from primary tumors and characterize their cellular and molecular features to find potential therapeutic drugs. The MGT cell lines demonstrated rapid cell proliferation and colony formation in an anchorage-independent manner. Vimentin and α-SMA levels were significantly elevated in MGT cell lines compared to normal canine kidney (MDCK) cells, while CDH1 expression was either significantly lower or not detected at all, based on quantitative real-time PCR (qRT-PCR) analysis. Functional annotation and enrichment analysis revealed that epithelial-mesenchymal transition (EMT) phenotypes and tumor-associated pathways, particularly the PI3K/Akt signaling pathway, were upregulated in MGT cells. BYL719 (Alpelisib), a PI3K inhibitor, was also examined for cytotoxicity on the MGT cell lines. The results show that BYL719 can significantly inhibit the proliferation of MGT cell lines in vitro. Overall, our findings suggest that the MGT cell lines may be valuable for future studies on the development, progression, metastasis, and management of tumors.

Genome-wide association study of mammary gland tumors in Maltese dogs.

Background: A genome-wide association study (GWAS) is a valuable tool for investigating genetic and phenotypic variation in many diseases. Objective: The objective of this study was to identify variations in the genomes of Maltese dogs that are associated with the mammary gland tumor (MGT) phenotype and to assess the association between each biological condition and MGT phenotype in Maltese dogs. Methods: DNA was extracted from 22 tumor samples and 11 whole blood samples from dogs with MGTs. Genome-wide single-nucleotide polymorphism (SNP) genotyping was performed, and the top 20 SNPs associated with various conditions and genetic variations were mapped to their corresponding gene locations. Results: The genotyping process successfully identified 173,662 loci, with an overall genotype completion rate of 99.92%. Through the quality control analysis, 46,912 of these SNPs were excluded. Allelic tests were conducted to generate Manhattan plots, which showed several significant SNPs associated with MGT phenotype in intergenic region. The most prominent SNP, located within a region associated with transcription and linked to the malignancy grade of MGT, was identified on chromosome 5 (p = 0.00001) though there may be lack of statistical significance. Other SNPs were also found in several genes associated with oncogenesis, including TNFSF18, WDR3, ASIC5, STAR, and IL1RAP. Conclusion: To our knowledge, this is the first GWAS to analyze the genetic predisposition to MGT in Maltese dogs. Despite the limited number of cases, these analyzed data could provide the basis for further research on the genetic predisposition to MGTs in Maltese dogs.

Modulating the GSH/Trx selectivity of a fluorogenic disulfide-based thiol sensor to reveal diminished GSH levels under ER stress.

We synthesized a fluorogenic disulfide-based naphthalimide thiol probe (ER-Naph) with a hydrophilic endoplasmic reticulum (ER)-guiding glibenclamide unit. Its ER targeting ability and high selectivity to GSH over thioredoxin, a potent competitor, were clearly demonstrated, both in solution and in vitro. Finally, a confocal microscopic investigation revealed that GSH levels in the ER were dramatically decreased under thapsigargin, brefeldin A, and tunicamycin-induced ER stress models.

Mycoplasma pneumoniae-associated Hepatitis without Lung Involvement.

Mycoplasma pneumoniae (M. pneumoniae) is a major cause of community acquired respiratory infections. And it also causes a number of extrapulmonary manifestations including cardiovascular, dermatological, musculoskeletal, and hematological systems. But, acute hepatitis without lung involvement is rare in adults. Here, we report a case of 32-year-old man who presented with fever, chilling, myalgia, and headache. Biochemical analysis showed severely impaired liver function and leukopenia. Laboratory tests and liver biopsy demonstrated a hepatocellular pattern of M. pneumoniae-associated acute hepatitis. Clinical symptoms and laboratory parameters are improved rapidly under treatment with macrolide. Therefore, We recommend that phsycians should consider a possibility of M. pneumoniae infection in acute hepatitis without lung and extrapulmonary involvement, when other more frequent causes have been excluded.

Clinical characteristics and progression of liver abscess caused by toxocara.

AIM: To evaluate the clinical characteristics and progression of liver abscess caused by toxocara. METHODS: We retrospectively reviewed the medical records of patients with serum IgG antibody to Toxocara canis and liver abscess diagnosed using abdominal computed tomography between February 2010 and February 2015. Among 84 patients exhibiting serum IgG antibody to Toxocara canis, 34 patients were diagnosed with liver asbscess and treated with albendazole. A follow-up period of 1 year was conducted. RESULTS: Mean patient age was 53 (34-79) years, with 26 (76.5%) patients being male. Twenty-one (61.7%) patients were moderate or heavy drinkers, 23 (67.6%) patients had a history of eating raw meat or liver and 6 (17.6%) patients owned pet dogs or cats. Main patient symptoms consisted of right upper quadrant pain, fever, and fatigue; 18 (52.9%) patients, however, presented with no symptoms. Lung involvement was detected in 444 (11.7%) patients. The eosinophil count increased in 29 (85.3%) patients at initial diagnosis, and decreased in most patients after albendazole treatment. The initial serum IgE level increased in 25 (73.5%) patients, but exhibited various response levels after albendazole treatment. Liver abscess formation improved in all patients. CONCLUSION: The liver abscess was improved with albendazole treatment.

[A case of IgG4-related sclerosing mesenteritis associated with Crohn's disease].

Sclerosing mesenteritis (SM) is a rare disease characterized by chronic nonspecific mesenteric inflammation and fibrosis of unknown etiology. Some tumefactive SM shows diffuse accumulation of IgG4-positive plasma cells and is considered as a part of the spectrum of IgG4-related disease. An association between inflammatory bowel disease and IgG4-related disease has been indicated. A 45-year-old woman visited our hospital due to weight loss with intermittent lower abdominal discomfort. Pelvic ultrasound revealed a mass-like lesion in the abdominal wall and pelvis MRI demonstrated a 5.9 cm sized wall-enhancing mass with heterogeneous signal intensity from right adnexa to the abdominal wall. Tumor resection and adhesiolysis was done because of severe adhesion with the small bowel, colon, bladder, uterus, and abdominal wall. Appendectomy was also performed due to adhesion and edematous change. Histological examination of the resected mass showed findings that were compatible with IgG4-related SM. The resected appendix showed chronic granulomatous inflammation without evidence of tuberculosis. She was diagnosed with Crohn's disease after undergoing colonoscopy and CT enterography. Herein, we report a rare case of IgG4-related SM that occurred in conjunction with Crohn's disease.

The differences in clinical aspect between specific language impairment and global developmental delay.

OBJECTIVE: To compare and analyze the clinical characteristics of children with delayed language acquisition due to two different diagnoses, which were specific language impairment (SLI, a primarily delayed language development) and global developmental delay (GDD, a language delay related to cognitive impairment). METHODS: Among 1,598 children who had visited the developmental delay clinic from March 2005 to February 2011, 467 children who were diagnosed with GDD and 183 children who were diagnosed with SLI were included in this study. All children were questioned about past, family, and developmental history, and their language competences and cognitive function were assessed. Some children got electroencephalography (EEG), in case of need. RESULTS: The presence of the perinatal risk factors showed no difference in two groups. In the children with GDD, they had more delayed acquisition of independent walking and more frequent EEG abnormalities compared with the children with SLI (p<0.01). The positive family history of delayed language development was more prevalent in children with SLI (p<0.01). In areas of language ability, the quotient of receptive language and expressive language did not show any meaningful statistical differences between the two groups. Analyzing in each group, the receptive language quotient was higher than expressive language quotient in both group (p<0.01). In the GDD group, the Bayley Scales of Infant Development II (BSID-II) showed a marked low mental and motor quotient while the Wechsler Intelligence Scale showed low verbal and nonverbal IQ. In the SLI group, the BSID-II and Wechsler Intelligence Scale showed low scores in mental area and verbal IQ but sparing motor area and nonverbal IQ. CONCLUSION: The linguistic profiles of children with language delay could not differentiate between SLI and GDD. The clinicians needed to be aware of these developmental issues, and history taking and clinical evaluation, including cognitive assessment, could be helpful to diagnose adequately and set the treatment plan for each child.

What is the real practice of exercise echocardiographic testing in asymptomatic patients with severe aortic stenosis?

BACKGROUND: Although exercise testing has been suggested to help predict clinical outcome, limited data are available to guide how exercise Doppler echocardiography (ECG) can be used clinically in asymptomatic patients with aortic stenosis (AS). The aim of this study was to assess the clinical value of exercise echocardiographic testing in asymptomatic patients with severe AS. METHODS: Symptom-limited treadmill exercise testing using the modified Bruce protocol was performed in 31 asymptomatic patients (mean age (62 ± 11) years) with severe AS (aortic valve area <1 cm(2), peak aortic velocity (AV Vmax) >4 m/sec, or a mean transaortic pressure gradient (AV mean PG) >40 mmHg (1 mmHg = 0.133 kPa)) with normal left ventricular (LV) systolic function (LV ejection fraction (EF) >50%). Clinical symptoms, vital signs, ECG, and Doppler hemodynamics were obtained during and/or immediately after exercise. RESULTS: Aortic valve replacement (AVR) was performed in 18 patients during follow-up. The patients who had AVR exhibited higher baseline AV mean PG (51 (35-84) vs. 44 (25.2-57.0) mmHg; P = 0.031). There were no significant differences between the AVR group and non-AVR group including exercise duration (7.47 (2.32-11.59) vs. 7.25 (4.06-10.52) minutes, P = 0.917), exercise capacity (10.1 (4.6-12.8) vs. 10.1 (7.0-12.8) metabolic equivalents, P = 0.675), and an increment in AV mean PG by exercise (18.5 (3.2-48.0) vs. 12.6 (4.4-32.1) mmHg, P = 0.366). Univariate regression analysis revealed that independent determinant of AVR was the baseline AV mean PG (P = 0.031). CONCLUSIONS: Although additional value of exercise ECG was demonstrated, baseline transaortic mean pressure gradient is the major determinant of AVR. Further large-scale prospective studies are required to determine whether surgery should be recommended in the presence of an abnormal exercise ECG in asymptomatic severe AS.

Contractile reserve determined on exercise echocardiography in patients with severe aortic regurgitation.

BACKGROUND: The aim of this study was to assess contractile reserve (CR) in patients with severe aortic regurgitation (AR), and potential clinical applications of CR in minimally symptomatic patients. METHODS AND RESULTS: Symptom-limited treadmill exercise echocardiography was performed in 58 asymptomatic or mildly symptomatic patients with chronic severe AR (male 69%, 50±14 years). Patients with increased ejection fraction (EF) after exercise were denoted as having CR (+) and those without increased EF were categorized as CR (-). CR (+) was found in 31 and CR (-) in 27 patients. Compared with CR (+), the CR (-) group had lower EF, larger effective regurgitant orifice, regurgitant volume, left ventricular (LV) end-diastolic dimension, LV end-systolic dimension (LVESD), and LV mass index. On multiple logistic regression analysis LVESD index (LVESDI; odds ratio -0.354, P<0.0001) was an independent predictor of CR. One-third of patients with LVESD >50mm, however, were found to be CR (+), while one-third of patients with smaller LVESD were CR (-). CONCLUSIONS: Although CR is best related to baseline resting LVESDI, one-third of patients were found to have discordance between the presence of CR and the LV dimension recommended for aortic valve replacement (AVR) in minimally symptomatic patients with severe AR, suggesting that exercise test may be able to further stratify the current guideline for AVR.

The importance of single trials: temporal and spatial resolution in event-related potential research.

In event-related potential (ERP) research, the average potential over many trials is a useful measure, but it obscures temporal variability across trials that may be functionally significant. Familiar ERP components have been shown to reflect the phase shifting of oscillatory activity that is present in varying degrees before the event. Even with single trials, understanding the underlying neural activity may require methods of dealing with the spatial superposition of neural signals at the head surface electrodes. We conclude with an illustration in which the modulation of averaged ERPs by trait anxiety may be related to underlying sources of theta oscillations.

Computational modeling of human head electromagnetics for source localization of milliscale brain dynamics.

Understanding the milliscale (temporal and spatial) dynamics of the human brain activity requires high-resolution modeling of head electromagnetics and source localization of EEG data. We have developed an automated environment to construct individualized computational head models from image segmentation and to estimate conductivity parameters using electrical impedance tomography methods. Algorithms incorporating tissue inhomogeneity and impedance anisotropy in electromagnetics forward simulations have been developed and parallelized. The paper reports on the application of the environment in the processing of realistic head models, including conductivity inverse estimation and lead field generation for use in EEG source analysis.

Randomized phase 2 study of irinotecan plus cisplatin versus gemcitabine plus vinorelbine as first-line chemotherapy with second-line crossover in patients with advanced nonsmall cell lung cancer.

BACKGROUND: The current study was performed to compare the nonplatinum-based combination of gemcitabine and vinorelbine (GV) with the combination of irinotecan and cisplatin (IP) as first-line chemotherapy with second-line crossover in patients with advanced nonsmall cell lung cancer (NSCLC). METHODS: Patients were randomly assigned to received either irinotecan at a dose of 65 mg/m(2) plus cisplatin at a dose of 30 mg/m(2) (Arm A) or gemcitabine at a dose of 900 mg/m(2) plus vinorelbine at a dose of 25 mg/m(2) (Arm B), each of which was administered on Days 1 and 8 every 3 weeks as the first-line therapy followed by crossover at the time of disease progression. RESULTS: A total of 146 patients were enrolled (75 patients in Arm A and 71 patients in Arm B); 138 patients were evaluable for tumor response and toxicity. During first-line therapy, IP was found to result in more grade 2+ nausea and vomiting (toxicity was graded according to the National Cancer Institute Common Toxicity Criteria [version 2.0]) (41% vs 12%; P = .0001) and alopecia (36% vs 10%; P = .0003). Pneumonitis was noted only with GV therapy (7% vs 0%; P = .058). During second-line therapy, IP was found to result in more grade 3 diarrhea (17% vs 2%; P = .039) and GV featured more cases of grade 3+ neutropenia (78% vs 40%; P = .0003). IP tended to generate more tumor responses (38% vs 26% as first-line therapy, and 30% vs 13% as second-line therapy) compared with GV. IP also demonstrated a favorable trend in median progression-free survival (4.6 months vs 3.8 months as first-line therapy and 4.5 months vs 2.6 months as second-line therapy) and overall survival (15.9 months vs 13.1 months; P = .3), but this difference was not statistically significant. The majority of patients who were refractory to IP also failed to respond to GV in the second-line setting. CONCLUSIONS: The platinum-based IP regimen appeared to be superior to the GV combination in terms of response rate. However, given the similar survival and better tolerability of the nonplatinum GV regimen, either treatment sequence would appear to be acceptable for the treatment of patients with advanced NSCLC.

Physiological Functionality and Enzyme Activity of Biomass from Pichia anomala Grown on Ginseng-Steaming Effluent.

A novel biomass was prepared from Pichia anomala KCCM 11473, which grew well in ginseng-steaming effluent (GSE), and its physiological functionalities and enzyme activities were determined. When the strain was cultured in the GSE (pH 6.0) at 30℃ for 48 h, 1.6 mg of biomass per ml-cultures was produced. The cell-free extract of the biomass showed high antihypertensive angiotensin I-converting enzyme inhibitory activity of 72.0% and anticholesteromia HMG-CoA reductase inhibitory activity of 46.5%. The cell-free extract also showed 13.0 U per ml and 8.5 U per ml of neutral protease activity and alkaline protease, respectively.

Effect of coping on endocrinoimmune functions in different stress situations.

The objective of this study was to examine the effects of coping strategies on the endocrine and immune functions in different stress situations. Thirty-eight medical students were enrolled in this study. Cell-mediated immune function was measured using the lymphocyte proliferative response to phytohemagglutinin (PHA) and interleukin-2 (IL-2) production during the nonexamination period and during the preexamination period. Endocrine functions were assessed by measuring the plasma levels of norepinephrine, adrenocorticotropic hormone (ACTH) and cortisol. The Global Assessment of Recent Stress (GARS) scale, the Stress Response Inventory, the anxiety, depression, and somatization subscales of the Symptom Checklist-90-revised, the Way of Coping-revised, the Toronto Alexithymia Scale and the Anger Expression Scale were used as psychometric measures. The subjects with higher levels of total GARS scores showed significantly higher IL-2 production during the nonexam period than those with lower levels of total GARS scores. During the same period, IL-2 production in the less positive reappraisal group was significantly higher than in the more positive reappraisal group. Lymphocyte proliferation in the group seeking less social support was also significantly higher than in the group seeking more social support. However, no significant association was found between the coping strategies and each of the hormone levels. These results suggest that positive reappraisal and seeking social support can be associated with the alteration of immune function during a chronic stress period. In particular, positive reappraisal is likely to reverse the stress-induced immune responses. This study did not find that neuroendocrine function such as the sympathetic-adrenal medullary axis or the hypothalamic-pituitary-adrenal axis is playing a mediating role in the relationship between coping and immunity.