PLASMA PHENYLALANINE DETERMINATION BY QUANTITATIVE DENSITOMETRY OF THIN LAYER CHROMATOGRAMS AND BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY IN RELATION WITH MODERN MANAGEMENT OF PHENYLKETONURIA.

BACKGROUND: The modern management of phenylketonuria (PKU) consists of generalized newborn screening (NBS) for hyperphenylalaninemia (HPA), confirmation of HPA in children detected in the NBS, introduction of dietary treatment in the first weeks of life, followed by monitoring the treatment of PKU for decades to maintain phenylalaninemia within the limits that will not affect the brain. The present study aimed to evaluate the usefulness of two chromatographic methodologies for determination of plasma Phe level in the routine management of PKU: the two dimensional thin layer chromatography (2D - TLC) and the high performance liquid chromatography (HPLC) procedures, respectively. MATERIAL AND METHODS: Samples of blood from 23 children with HPA detected by neonatal screening or with confirmed PKU who received treatment by low-Phe diet were analyzed to estimate the plasma Phe level by the two chromatographic procedures. RESULTS: In case of three subjects the very low concentrations of plasma Phe could not be detected by the 2D - TLC methodology, since the spot was not visible on the chromatogram. In four patients the differences between the values of plasma Phe determined by the two methodologies are not statistically significant, while in fifteen subjects the differences are highly statistically significant. This is due to the greater errors that appear in the case of 2D - TLC methodology. In the range of concentrations of plasma Phe higher than 360 µmol/L (which is the cut-off value for HPA), although in four cases there were statistically significant differences in the level of plasma Phe determined by the two methodologies, the value obtained by the 2D - TLC methodology was high enough to influence the decision of changing the diet so that HPA is kept under control. In addition, the intense spot of Phe on the 2D - TLC chromatogram may be detected even by un unexperienced laboratory specialist. CONCLUSION: The HPLC procedure for measurement of plasma Phe level is very suitable to be used in the routine management of PKU. The 2D - TLC procedure may be accompanied by relatively high errors; however, it detects patients with severe PKU.

Credibility elements of eWOM messages in the context of health care services. A Romanian perspective.

The purpose of this paper is to investigate the Romanian consumers' determinants of eWOM messages' perceived credibility in the context of health care services. We selected a sample of 127 women and we administered a questionnaire. We used the partial least squares to uncover the established relationships between the variables of the model, namely the argument strength and the source credibility of a eWOM message and the intention to purchase a health care service based on the information embedded in the eWOM messages. The results revealed that all variables had positive direct correlations with each other but the argument strength of a message has the highest impact on the intention.

Classification of breast carcinomas according to gene expression profiles.

Breast carcinomas represent an important health problem. Understanding the development of breast cancer from precursor is critical for clinical treatment and prevention, however little is known about the molecular events involved in the progression to cancer. The advent of gene expression microarray technology provides a new powerful tool to assist in the determination of diagnosis, prognosis and treatment. In this paper, we present the recent DNA microarray studies that describe how gene expression profiling is being used to classify specimens of breast carcinomas based on molecular properties of the tumor and to identify gene expression patterns related to clinical outcome. In present, data are available that show that gene expression profiles can be used to distinguish cell type-specific gene clusters (stromal, epithelial, mesenchymal and proliferation status) and to classify breast tumors as basal-like, luminal-like, ERBB2 overexpressing and normal breast-like. Profiles associated with good prognosis and poor prognosis of young axillary node negative patients have been identified. The microarray technology will become in the near future a molecular complement to histopathology and immnuhistochemistry.

Optimizing the technological and informational relationship of the health care process and of the communication between physician and patient--factors that have an impact on the process of diagnosis from the physician's and the patient's perspectives.

OBJECTIVE: The optimization of a diagnosis process and fluency in the Health Care sector in Romania. A key to discover this complex process was to determine a correlation between the physicians and the use of information technology, on one side and the patients' perspective on the other. HYPOTHESIS: Integrating information technology in a physician's activity will lead to lower costs and less time spent while diagnosing patients. Using the electronic medical records and introducing a unified database with the patients' medical histories will make the process of diagnosis easier. METHODS: We studied the diagnosis from the point of view of 304 patients in a public hospital and 320 physicians working there. RESULTS: We believed that time and accessibility to different physicians makes the diagnosis process a burden for a patient and implicitly lead to dissatisfaction with health care services. We supposed that the burden of diagnosis for physicians comes from the lack of Internet connection and computer usage knowledge. We have found out that most physicians know how to use the computer at an intermediate level and have access to Internet, online journals and databases and do not use emails to a higher extent to communicate to other specialists, but do not rely entirely on the electronic medical records. Most physicians think that it is not technology, which stands in the way of proper and fast diagnosis but the financing and the paper work from the Romanian health system. Solutions that might be taken into account to entirely motivate physicians to use electronic medical records are: 1. Adjustments can be made to the computer software interface in order to make the design more consistent (to eliminate the paper forms) and user friendly. 2. Physicians can be provided with more training and knowledge. DISCUSSION: After some statistical tests have been applied to find a correlation between the chosen variables, we have reached the conclusion that the results are encouraging and there is no correlation between the degree of the impact of Preventive Medicine and the healthy behavior of the respondents.

Optimizing the technological and informational relationship of the health care process and of the communication between physician and patient. The impact of Preventive Medicine and social marketing applied in Health Care on youth awareness- preliminary study.

RATIONALE: In this case study we wanted to find out the impact of Preventive Medicine and implicitly social marketing upon young students with the average age of 19, belonging to the academic environment in Romania. METHODS: The study lasted one month and consisted of a questionnaire that was conceived and applied to 304 adolescents. The questionnaire contained demographic and personal information, such as age, origin, gender, marital status and some questions related to the respondents' attitude towards several issues that are inserted in the preventive medicine discipline, such as the date of their last consultation, if the respondents were registered to a family physician, suffered from chronic diseases, what was the rate of doing physical exercises, if they ate salty and fat meals, if they were on a diet, their rate of alcohol, caffeine and tobacco consumption. RESULTS: The panel was made up of more female respondents than male, with the average age rate of 19, who had medical consultations in the last 3 months, are included in the evidence of a family physician, had no chronic diseases, usually do workout exercises moderately, are not on a diet and have 3 meals per day. The meals are medium salty and rarely rich in fats. They drink 2 cups of tea per day and are non-alcohol drinkers and non-smokers. DISCUSSION: After applying several statistical tests to find a correlation between our variables, we reached the conclusion that even if the results are encouraging; there is no correlation between the impact of Preventive Medicine and the respondents' health behavior.

Statistical correlations between peripheral blood lymphocyte subpopulations and tumor inflammatory infiltrate in stage I of skin melanoma.

The article presents statistical correlations of immune cell parameters investigated in patients diagnosed with skin melanoma stage I. Recent data indicate a suppressed immune response, probably sustained by immune-regulating molecules expressed or shed from the tumor. These molecules block an efficient immune response and thus the tumor develops. All the molecules that are part of the tumor escape mechanisms can be targets for immune-mediated anti-tumor agents. We try to find the significance of some immunohistochemical markers (UCHL1, CD4, CD8) in tumoral inflammatory infiltrate and to establish the statistical correlations between molecular markers and tumor grade and stage. The studied parameters were: CD3+, CD4+, CD8+, CD56+16+ and CD19+. The statistical results were performed with SPSS v. 15.0. We demonstrate that a CD4+ on-going immune response is elicited in the investigated patients. We found a possible compensatory mechanism between T-lymphocytes and NK-cells and also between the antibodies generating cells and the natural cytotoxic cells. We are confident that these statistical correlations between clinical, immunological and immunohistochemical data can be useful in the disease management and personalized immune-therapy.

Quantitative volumetric MRI study of the cerebellum and vermis in schizophrenia: clinical and cognitive correlates.

OBJECTIVE: Recent evidence suggests that the cerebellum may play a role in higher cognitive functions and, therefore, may play an important role in schizophrenia. METHOD: The authors used magnetic resonance imaging to measure cerebellum and vermis volume in 15 patients with schizophrenia and 15 normal comparison subjects. RESULTS: They found that 1) vermis volume was greater in patients with schizophrenia than in normal subjects, 2) greater vermis white matter volume in the patients with schizophrenia significantly correlated with severity of positive symptoms and thought disorder and with impairment in verbal logical memory, and 3) patients with schizophrenia showed a trend for more cerebellar hemispheric volume asymmetry (left greater than right). CONCLUSIONS: These data suggest that an abnormality in the vermis may contribute to the pathophysiology of schizophrenia.

Study on the prevalence of eccentric and concentric atherosclerotic plaques in various segments of the coronary arterial tree, in unselected and selected Bucharest population samples.

A study was carried out on the prevalence and distribution of eccentric and concentric atherosclerotic plaques in the coronary arterial tree of 916 unselected apparently healthy subjects who died from accidents and of 150 selected patients who died from coronary heart disease. The results show that the percentages of eccentric and concentric plaques vary with age, sex, anatomical branching pattern and cause of death; there are also differences between major coronary arteries and main branch vessels. In addition to eccentric and concentric plaques, we revealed the existence of mixed (both eccentric and concentric) plaques, mainly located at branch points.

Prevalence of thick intimas and of obstructive lesions in the vessels supplying the conduction system of the heart.

A study of the prevalence of thick intimas and of obstructive lesions in the vessels supplying the conduction system of the heart (first septal artery, sinus node artery, atrioventricular node artery, posterior descending artery) was carried out on both non-hospitalized and hospitalized subjects. A total of 932 cases aged 6 to 55 years were investigated. The age period of the onset of both intimal thickenings and atherosclerotic plaques, the percent of cases with intimal thickenings and atherosclerotic plaques in successive age groups and the highest value of the intima thickness/media thickness ratio, were recorded. Comparative data are presented between apparently healthy subjects who died of accidental causes and patients of similar age and sex, who died of coronary heart disease. Sudden cardiac death cases are presented showing as the most important lesion the obstruction of the sinus node and atrioventricular node arteries. The results point to a need for a more frequent routine examination of the vessels supplying the conduction tissue of the heart.

Role of thrombosis in the onset, progression and obstructive character of coronary atherosclerotic plaques.

A study was carried out on the role of successive microthrombi incorporation and intramural thrombi formation in the onset, progression and obstructive character of coronary atherosclerotic plaques. The coronary arterial tree of 836 apparently healthy subjects 1 to 50 years old who died of accidental causes was investigated. The onset of coronary atherosclerotic plaques having as precursors thrombi could be detected in 10% of advanced lesions. The left anterior descending artery appeared as the vessel of the coronary arterial tree the most susceptible to plaque development induced by thrombi and to the occurrence of thrombi on preexisting plaques. This occurrence of thrombi on preexisting plaques was recorded in 48% of subjects aged 46 to 50 years and was followed by a severe luminal narrowing in the major coronary arteries.

A further study on the influence exerted by vascular geometry on coronary atherosclerotic involvement.

A further study was carried out on the relationship between coronary artery geometry and atherosclerotic involvement. The results show that: a) the coronary artery geometry may expose an individual at an early age to an increased risk for developing atherosclerotic lesions; b) the presence, in the coronary arterial bed of vessels with large diameters (greater than 2 mm) and the succession of numerous branching points also appeared as atherogenic conditions; c) the type of atherosclerotic lesion (proliferative, insudative, necrotic, lipid-rich) was, in our material strongly influenced by the geometry of branch sites.

Prevalence of the "normal" aspect of the coronary arteries in an unselected population sample of Bucharest aged 1-60 years.

The coronary arterial trees obtained from 640 subjects 1-60 years old were studied grossly and by light microscopy, including morphometric methods. The highest values of of the ratio intima/media thickness were recorded in successive age groups, with special reference to those topographic sites where this ratio acquired an abnormal value. The percent of children, adolescents, young and mature adults free of atherosclerotic plaques, gelatinous lesions and intima necrotic areas, fatty streaks and intramural thrombi was also evaluated. An association was revealed between the number of subjects with anatomically normal coronary arteries and the presence of the common type of distribution of the coronary arteries in these subjects.

Atherogenic anatomical patterns of coronary arteries branching related to cardiac and non-cardiac causes of death.

A gross and light microscopic study carried out on 102 selected male subjects aged 51-70 who died of ischemic heart disease and on 102 selected, male subjects of similar age, who died of noncardiac causes revealed that: in subjects who died of ischemic heart disease the prevalence of atherogenic deviations from the common type of distribution of the coronary arteries was 2-4 times greater than in subjects who died of non-cardiac causes; subjects who died of ischemic heart disease, with the atherogenic deviations from the common tape of distribution of the coronary arteries showed an increased number of stenotic atherosclerotic plaques and a particular propensity to "clustering" of these lesions, predominantly around branch mouths.

A method which improves anatomo-clinical correlations in ischemic heart disease.

A study carried out on 595 unselected subjects aged 21-55 years who had died of accidental causes and on 56 selected patients who had died of myocardial infarction showed that the light microscopic examination of coronary branch vessels led to an obvious improvement of anatomo-clinical correlations in ischemic heart disease. These branch vessels included atherosclerotic plaques in 51% of unselected subjects 51-55 years old, dead of accidents and in 73% of selected patients dead of myocardial infarction. Severe stenotic plaques of possible clinical significance were found in 14% of subjects dead of accidents and in 48% of patients dead of myocardial infarction. In these cases the atherosclerotic involvement in the major coronary arteries (left anterior descending, circumflex and right coronary arteries) did not reach the level of possible clinical significance (greater than or equal to 75% reduction of the vessel diameter). Several examples emphasize the importance of stenotic lesions in the first diagonal, first septal, posterior descending, left marginal and right marginal vessels, as well as in the vessels supplying the sinoatrial and atrioventricular nodes for an adequate anatomo-clinical correlation in ischemic heart disease.

Age and atherosclerosis--related changes occurring in the "intercalated vascular segments" of human coronary arteries.

The term "intercalated vascular segment" is used by the authors to designate a structural accomodation to the existence of diffuse thickened intimas, occurring as local vascular connections between the lumen of the main coronary arteries and the branch mouths of the daughter vessels. They exhibit the light microscopic feature of cylindrical conduits including only an endothelial sheet encased by the intimal connective tissue. The most important change related to aging and atherosclerotic involvement seems to be the conversion of some intercalated vascular segments in rigid, undistensible and narrowed conduits; this might produce local changes similar to those of a fixed stenosis. In the material examined the presence of narrowed conduits was sometimes associated with the existence of small areas of necrosis in the myocardium supplied by the vessel corresponding to the respective intercalated vascular segment.