Mutation analysis of candidate genes in melanoma-prone families: evidence of different pathogenetic mechanisms at chromosome 9P21

Putative tumour suppressor genes CDKN2A and CDKN2B (on chromosome 9p21) and CDKN2A-interacting cell growth regulatory genes CDK4 and Id-1 have been demonstrated to be involved in the pathogenesis of malignant melanoma (MM). Mutation analysis of these candidate genes was performed in MM families from southern Italy with three or more affected members or two affected members and one or more relative with histologically diagnosed atypical naevus. Two CDKN2A mutations, Arg24Pro and 1-292 G>A, were observed in two (15%) families; except for CDKN2A and Id-1 polymorphisms, no sequence variations were detected in the remaining genes. Screening among 119 sporadic MM cases revealed two additional CDKN2A mutations at very low prevalences. Identification of a large shared haplotype at 9p21 in some MM families negative for CDKN germline mutations suggests that other CDKN-inactivating mechanisms may be responsible for MM predisposition or, alternatively, additional susceptibility gene(s) may be present on chromosome 9p21. Fluorescence in situ hybridization analysis of a subset of MM tissue sections seemed to indicate that the D9S171 locus may be involved in MM pathogenesis.     

Localization of syntactic and semantic brain responses using magnetoencephalography

Electrophysiological methods have been used to study the temporal sequence of syntactic and semantic processing during sentence comprehension. Two responses associated with syntactic violations are the left anterior negativity (LAN) and the P600. A response to semantic violation is the N400. Although the sources of the N400 response have been identified in the left (and right) temporal lobe, the neural signatures of the LAN and P600 have not been revealed. The present study used magnetoencephalography to localize sources of syntactic and semantic activation in Finnish sentence reading. Participants were presented with sentences that ended in normally inf lected nouns, nouns in an unacceptable case, verbs instead of nouns, or nouns that were correctly inflected but made no sense in the context. Around 400 msec, semantically anomalous last words evoked strong activation in the left superior temporal lobe with significant activation also for word class errors (N400). Weaker activation was seen for the semantic errors in the right hemisphere. Later, 600-800 msec after word onset, the strongest activation was seen to word class and morphosyntactic errors (P600). Activation was significantly weaker to semantically anomalous and correct words. The P600 syntactic activation was localized to bilateral sources in the temporal lobe, posterior to the N400 sources. The results suggest that the same general region of the superior temporal cortex gives rise to both LAN and N400 with bilateral reactivity to semantic manipulation and a left hemisphere effect to syntactic manipulation. The bilateral P600 response was sensitive to syntactic but not semantic factors.     

Higher Serum Uric Acid on Admission Is Associated with Higher Short-term Mortality and Poorer Long-term Survival After Myocardial Infarction: Retrospective Prognostic Study

Aim

To assess serum uric acid (SUA) levels determined on admission as a potential predictor of short-term mortality and long-term survival in acute myocardial infarction (AMI) patients.

Method

Data for this retrospective prognostic study were drawn from the patient database of the Varaždin County General Hospital in Varaždin, Croatia. We included consecutive patients with verified AMI admitted within 48 hours since the symptom onset during the period between January 1, 1996 and December 31, 2001. Long-term survival/mortality data were collected through direct contacts with patients and search of the community death registries. Relative risks (RR) and hazard ratios (HR) by 10 µmol/L increase in SUA were determined using modified Poisson regression with robust error variance and proportional hazard regression, respectively.

Results

A total of 621 patients (age 27-90 years, 64.7% men, 77.5% AMI with ST elevation, SUA 63-993 µmol/L) were included. Higher SUA on admission was independently associated with higher in-hospital mortality (RR, 1.016; 95% confidence interval [CI], 1.001-1.031, P = 0.043) and higher thirty-day mortality (RR, 1.016; 95% CI, 1.003-1.029, P = 0.018). Considered covariates were demographics, pre-index event cardiovascular morbidity and treatment, on-admission serum creatinine, total cholesterol and triglycerides, AMI characteristics, and peak creatine phosphokinase. Higher SUA on admission was also independently associated with poorer long-term survival (ie, higher all-cause mortality) (HR, 1.105; 95% CI, 1.020-1.195, P = 0.010). Considered covariates were demographics, laboratory variables on admission, AMI characteristics, peak creatine phosphokinase, acute complications, and treatment at discharge.

Conclusion

Higher serum uric acid determined on admission is associated with higher in-hospital mortality and thirty-day mortality and poorer long-term survival after AMI.In humans, uric acid (UA) is the end product of purine catabolism (1). Its serum levels (SUA), governed by the production (liver) and elimination (mainly the kidney) rates, are influenced by genetically determined factors (eg, activity of synthesizing enzymes or renal transporter systems), racial and demographic characteristics (eg, sex, gonadal function in women, obesity), diet (eg, purine-rich foods, fructose, alcohol), habits (eg, SUA is lower in smokers and increases after quitting), morbidity (eg, heart or renal failure, malignancies), and medications (eg, diuretics, cytotoxic agents) (1-4). The role of SUA in cardiovascular and renal diseases has been intensively investigated, although not without controversy (5). On the molecular and cellular level, UA exerts a number of effects of potential interest: it is one of the most important antioxidants in plasma, but at high concentrations it may promote oxidative stress; it may induce endothelial dysfunction and vascular smooth muscle cell proliferation in vitro, platelet aggregation, and microinflammation; increased UA causes tubulointerstitial inflammation, morphological and functional changes in the glomeruli and renal arteriole and increased salt sensitivity (3,5). There is now sufficient evidence to consider increased SUA as an etiological factor in “hyperuricemic hypertension” or “salt-sensitive kindey-dependent hypertension” (3,5). Clinical and epidemiological studies have linked increased SUA to occurrence and outcomes of diabetes mellitus, metabolic syndrome, and chronic renal failure (3,5,6). It has also been suggested as a risk factor for occurrence and a predictor of poorer outcomes in acute stroke (7-9) and a risk factor for occurrence/outcomes in various aspects of cardiovascular morbidity (3,5,6). However, there are also views that SUA is not relevant in the pathophysiology of cardiovascular diseases and that it should be viewed as a secondary side-marker of etiologically relevant processes (3,5).Acute myocardial infarction (AMI) is the most dramatic manifestation of the coronary artery disease (CAD) (10). High SUA has been indicated as a risk factor for CAD (10) and as an independent prognostic factor of poorer outcomes (occurrence of AMI, fatal AMI, sudden death, all-cause mortality) in patients with verified CAD (11,12). Less is known about SUA as a potential prognostic/risk factor for outcomes in patients affected specifically by AMI. A recent retrospective analysis from Japan (13) observed a univariate association between higher SUA on admission (within 48 hours since the symptom onset) and higher thirty-day mortality (fourth vs first quartile SUA values) in AMI patients. It also reported an independent association between higher SUA and poorer long-term survival (13). Having in mind potential ethnic/racial specificities and cultural differences (eg, diet, alcohol consumption), we aimed to investigate SUA levels determined on admission as a potential predictor of short-term mortality (while accounting for relevant covariates) and long-term survival in a sample of patients of European descent (Caucasians) with verified AMI.     

A Novel Organotypic Model Mimics the Tumor Microenvironment

Carcinoma cell invasion is traditionally studied in three-dimensional organotypic models composed of type I collagen and fibroblasts. However, carcinoma cell behavior is affected by the various cell types and the extracellular matrix (ECM) in the tumor microenvironment. In this study, a novel organotypic model based on human uterine leiomyoma tissue was established and characterized to create a more authentic environment for carcinoma cells. Human tongue squamous cell carcinoma cells (HSC-3) were cultured on top of either collagen or myoma. Organotypic sections were examined by immunohistochemistry and in situ hybridization. The maximal invasion depth of HSC-3 cells was markedly increased in myomas compared with collagen. In myomas, various cell types and ECM components were present, and the HSC-3 cells only expressed ECM molecules in the myoma model. Organotypic media were analyzed by radioimmunoassay, zymography, or Western blotting. During carcinoma cell invasion, matrix metalloprotease-9 production and collagen degradation were enhanced particularly in the myoma model. To evaluate the general applicability of the myoma model, several oral carcinoma, breast carcinoma, and melanoma cell lines were cultured on myomas and found to invade in highly distinct patterns. We conclude that myoma tissue mimics the native tumor microenvironment better than previous organotypic models and possibly enhances epithelial-to-mesenchymal transition. Thus, the myoma model provides a promising tool for analyzing the behavior of carcinoma cells.Tumor growth and invasion are not just determined by the malignant tumor cells, but instead various cell types and the extracellular matrix (ECM) of the tumor microenvironment affect the outcome.¹ Particularly, fibroblasts have many prominent roles in the cancer progression. In fact, in many carcinomas, the majority of the stromal cells are fibroblasts that possess myofibroblastic characteristics and are called cancer-associated fibroblasts. They produce ECM molecules, proteases, growth factors, and chemokines that crucially affect the carcinoma cell behavior.^2,3 In this context, the organotypic three-dimensional skin model developed by Fusenig et al⁴ replicates the in vivo situation more closely in vitro than the two-dimensional cell culture experiments. The model allows studying of carcinoma cell invasion in three-dimensional collagen gel embedded with fibroblasts. The degree of invasion can also be quantitatively analyzed.^5,6 However, this kind of organotypic model remains somewhat artificial due to the lack of other cell types besides fibroblasts and ECM components that are present in vivo. In addition to the carcinoma cells and fibroblasts, endothelial and inflammatory cells, as well as several ECM molecules, are known to contribute to the tumor growth. The induction of angiogenesis, recruitment of inflammatory cells, and increased turnover of ECM components result in tumor progression.^7,8 Therefore, we wished to determine whether real human tissue can be used in the organotypic method to provide a more natural stroma-like environment for studying carcinoma cell invasion. We used uterine leiomyoma tissue, which mainly consists of smooth muscle actin (SMA)-positive cells and collagens.⁹ The existence of various additional cell types and proteins in the myoma tissue was characterized, and the invasiveness of malignant human tongue squamous cell carcinoma cells (HSC-3) into this novel myoma organotypic culture was measured by different methods and compared with the traditional collagen organotypic model. To test the general applicability of the myoma model, the invasion patterns of various cell lines were examined in myoma and collagen organotypic cultures.     

Review of computer vision application in in vitro fertilization: the application of deep learning-based computer vision technology in the world of IVF

Journal of Assisted Reproduction and Genetics - In vitro fertilization has been regarded as a forefront solution in treating infertility for over four decades, yet its effectiveness has remained...     

Coeliac disease autoantibodies in seminal plasma from cases with screen-detected coeliac disease

Morphological changes in the remnant mucosa 2–20 years after operation for duodenal ulcer (DU) were analyzed in 124 male subjects by means of stochastic mathematics. Fifty non-operated male DU patients and 168 age-matched males from a population sample served as controls. Substantially no progression of gastritis was found in the DU patients. In contrast, the operative intervention caused a rapid initial progression of gastritis, continuing after 2 years approximately at the same speed as in the population at large.     

Typing of Human Campylobacter jejuni Isolates in Finland by Pulsed-Field Gel Electrophoresis

A total of 69 pulsed-field gel electrophoresis (PFGE) types were identified among 176 Campylobacter jejuni isolates from Finnish patients. In two geographic areas studied, five predominant PFGE types comprised over 40% of the isolates. One-third of the isolates had unique PFGE types. In small outbreaks, identical PFGE patterns were demonstrated, indicating a common source of infection.