A novel strategy based on histological protein profiling in‐silico for identifying potential biomarkers in urinary bladder cancer

OBJECTIVE

To screen a publicly available immunohistochemistry (IHC) based web‐atlas, to identify key proteins in bladder cancer that might serve as potential biomarkers.

MATERIALS AND METHODS

The first version of the Human Protein Atlas (HPA 1.0), with 660 proteins, was visually examined to identify proteins with a variable staining pattern among the 12 tissue samples representing bladder cancer. None or limited previous characterization in bladder cancer, as well as a supportive Western blot, were also required. The selected proteins were then evaluated in an independent set of patient samples (106 tumour samples of differing stage and grade) represented in a tissue microarray (TMAi). The IHC expression of the identified proteins in the TMAi was scored and related to tumour stage and grade.

RESULTS

The expression profiles of the 13 proteins selected from the web‐atlas were confirmed in the TMAi. Expression patterns for seven proteins were significantly altered (P < 0.05) with higher stage and/or grade. Three of those (CN130, DSG3, PHF6) lack characterization in bladder cancer, whereas the remaining four proteins have previously been suggested as key proteins/potential biomarkers in cancer, some of them also in bladder cancer.

CONCLUSION

New candidate proteins for urinary bladder cancer were identified through screening of the publicly available HPA 1.0. Although further evaluation is necessary, this strategy is promising in the search for new biomarkers, with potential to improve the management of patients with this disease.     

Aggression and Withdrawal Related Behavior within Conflict Management Progression in Preschool Boys with Language Impairment

Objective This study examined conflict behavior in naturalistic preschool settings to better understand the role of non-affiliative behavior and language in conflict management. Method Free-play at preschool was filmed among 20 boys with typically developing language (TL) and among 11 boys with Language Impairment (LI); the boys 4–7 years old. Conflict behavior was coded and analyzed with a validated system. Post-conflict non-affiliative behavior (aggression and withdrawal) displays, and the links between the displays and reconciliation (i.e., former opponents exchange friendly behavioral shortly after conflict termination) was examined. Results Group comparisons revealed boys with LI displayed aggression in a smaller share of conflicts, but exhibited ‹active’ withdrawal (left the room), in a larger conflict share. Boys with TL overcame aggression (more common TL behavior) and after reconciled, to a greater extent than the boys with LI after active withdrawal (more common LI behavior). Also, after reciprocal or only verbal aggression, boys with LI reconciled to a lesser extent than boys with TL. Conclusions The boys with LI demonstrated difficulties confronting conflict management, as well as concluding emotionally heightened and aggressive behavioral turns.     

Chronic hyperglycemia but not glucose variability determines HbA1c levels in well-controlled patients with type 2 diabetes

To determine the relationships between HbA1c, characteristics of hyperglycemia and glycemic variability in well-controlled type 2 diabetes (HbA1c<7.0%), we studied 63 primary-care patients (36 men and 27 women), aged 34-75 years, with type 2 diabetes for 2-32 years using a continuous glucose monitoring system (CGMS) and standardized meal test (MMT). Duration of hyperglycemia (>8.0 mmol/l), standard deviation score (S.D.-score) and mean amplitude of glycemic excursions (MAGE) were analyzed from CGMS data and postprandial glucose during MMT (PPG(MMT)). Patients were hyperglycemic for 5.7h/day (median), experienced 4.1 hyperglycemic episodes/day, and 78% exceeded PPG levels of 8.0 mmol/l. HbA1c, though associated with the extent of hyperglycemia (r=0.40, p<0.001), failed to correlate with S.D.-score and MAGE. Multiple regression analysis demonstrated that HbA1c was predicted only by fasting glucose (R(2)=0.24, p<0.001) but neither by PPG(MMT), duration of hyperglycemia, S.D.-score nor MAGE. CGMS and meal test provide the tools for complete characterization of glycemia in type 2 diabetes. In well-controlled type 2 diabetes, HbA1c correlates with chronic hyperglycemia but not with glucose variability. Our data suggest that chronic sustained hyperglycemia and glucose fluctuations are two independent components of dysglycemia in diabetes.     

Asthma in the elderly

A population aged 65 and over has been increasing in the developed countries. The prevalence of asthma in elderly patients is estimated between 6.5 and 17%. Asthma is an important cause of morbidity and mortality in the elderly. Moreover, death due to asthma occurs mostly in elderly patients. Only a few studies have reported the characteristics of asthma in the elderly patients. Two distinct clinical presentations of asthma have been described in the elderly. There are differences both in the pathophysiology and the clinical manifestation of asthma between elderly patients with a long-standing disease and those with late-onset disease. Additionally, aging of the respiratory system influences the asthma presentation. Asthma has been presented for many years may lead to persistent obstructive ventilatory defect and can mimic chronic obstructive pulmonary disease. Irreversible obstruction is commonly observed in elderly patients with asthma. The differential diagnosis of asthma is difficult in older adults and asthma is underrecognized and undertreated in the older population. Undertreatment is common in elderly asthmatics which largely is related to diagnostic issues.     

Poor prognosis in carcinoma is associated with a gene expression signature of aberrant PTEN tumor suppressor pathway activity

Pathway-specific therapy is the future of cancer management. The oncogenic phosphatidylinositol 3-kinase (PI3K) pathway is frequently activated in solid tumors; however, currently, no reliable test for PI3K pathway activation exists for human tumors. Taking advantage of the observation that loss of PTEN, the negative regulator of PI3K, results in robust activation of this pathway, we developed and validated a microarray gene expression signature for immunohistochemistry (IHC)-detectable PTEN loss in breast cancer (BC). The most significant signature gene was PTEN itself, indicating that PTEN mRNA levels are the primary determinant of PTEN protein levels in BC. Some PTEN IHC-positive BCs exhibited the signature of PTEN loss, which was associated to moderately reduced PTEN mRNA levels cooperating with specific types of PIK3CA mutations and/or amplification of HER2. This demonstrates that the signature is more sensitive than PTEN IHC for identifying tumors with pathway activation. In independent data sets of breast, prostate, and bladder carcinoma, prediction of pathway activity by the signature correlated significantly to poor patient outcome. Stathmin, encoded by the signature gene STMN1, was an accurate IHC marker of the signature and had prognostic significance in BC. Stathmin was also pathway-pharmacodynamic in vitro and in vivo. Thus, the signature or its components such as stathmin may be clinically useful tests for stratification of patients for anti-PI3K pathway therapy and monitoring therapeutic efficacy. This study indicates that aberrant PI3K pathway signaling is strongly associated with metastasis and poor survival across carcinoma types, highlighting the enormous potential impact on patient survival that pathway inhibition could achieve.     

Chronic treatment with sildenafil stimulates Leydig cell and testosterone secretion

The phosphodiesterase type 5 (PDE5) inhibitor, Sildenafil, is a novel, oral treatment approach for pulmonary hypertension. As Leydig cells present PDE5, this study was conducted to investigate the effects of the chronic treatment with Sildenafil (25 mg/kg) on male Swiss Webster mice steroidogenesis. After a 4-week long experimental design, Leydig cells were analysed by morphological and immunocytochemical procedures. Serum testosterone was assayed by radioimmunoassay. Leydig cells presented noteworthy ultrastructural alterations, such as a vesicular smooth endoplasmic reticulum, large vacuoles scattered through the cytoplasm, enlarged mitochondria with discontinue cristaes and whorle membranes with vesicles at the periphery, which are typical characteristics of an activated steroid-secreting cell. Important immunocytochemical labelling for steroidogenic acute regulatory protein, cytochrome P450 side-chain cleavage enzyme and testosterone were detected in isolated Leydig cells. In addition, Sildenafil-treated mice showed significant increased levels of total testosterone. The results obtained in the present study are consistent with the hypothesis that the accumulation of cyclic guanosine monophosphate by PDE5 inhibition could be involved in the androgen biosynthesis stimulation. Important clinical implications of hormonal disorders should be taken into account for patients with pulmonary hypertension.     

Neuroimaging as a tool for pain diagnosis and analgesic development

Neuroimaging makes it possible to study pain processing beyond the peripheral nervous system, at the supraspinal level, in a safe, noninvasive way, without interfering with neurophysiological processes. In recent years, studies using brain imaging methods have contributed to our understanding of the mechanisms responsible for the development and maintenance of chronic pain. Moreover, neuroimaging shows promising results for analgesic drug development and in characterizing different types of pain, bringing us closer to development of mechanism-based diagnoses and treatments for the chronic pain patient.     

Use and accidental exposure to hallucinogenic agents reported to the Czech Toxicological Information Centre from 1995 to 2008

Intoxication by hallucinogenic agents is relatively common in the Czech Republic, with most cases of the use of these agents being by adolescents and young people. The objective of the study was to evaluate the number, trends, gender and age of the subjects, and the severity of exposure in intoxication by hallucinogenic plant and mushroom substances, in comparison with synthetic drugs, brought to the attention of the Toxicological Information Centre (TIC) through inquiries over the past 14 years (1995-2008) (from total 3,702 calls concerning the use of both natural and synthetic drugs, 917 calls were due to the use of hallucinogenic plants and mushrooms).