Application of experimental design to the development of an HPLC method for the analysis of ochratoxin A in Triticum aestivum grain.

Ochratoxin A is a mycotoxin, a natural product of Aspergillus and Penicillium species. It can be present in grain from Triticum aestivum, (Graminae) and other starch-abundant cereals. This paper describes the investigation of ochratoxin A in grain from Triticum aestivum using a statistically optimized HPLC method. The assay was developed using two mathematical statistical models: factorial design and response surface mapping. The final step was to optimize the values of variables by response surface design. The analysis of variance 'ANOVA' method was applied to the analytical results in order to construct an adequate model. The optimal experimental conditions obtained by the response surface diagram method were: pH = 2.5, composition of the mobile phase acetonitrile: water 55:45 v/v and flow rate 1.0 ml/min. with a C18 column. Retention time and capacity factor for ochratotoxin A were 7.46 min. and 1.19, respectively.

[Addison's disease]. / Adisonova bolest.

This is a report on a patient 57 years old, hospitalized because of expressed fatigue, nausea, vomiting and epigastrium pain. Hyperpigmentation of skin and mucous and hypotension were present; and hypocorticism was suspected while ex iuvantibus substitutional therapy of corticosteroides was introduced. By testing thyroid gland function, hypothyroidism was established, so we suspected the presence of united pluriglandular insufficiency--Schmidt Syndrome. The patient reacted well to the substitutional therapy of hydrocortisone and levothyroxine. The diagnosis has been confirmed at the Clinic for Endocrinology in Novi Sad and today the patient has regular checks at the outpatients clinic for endocrinology in Sombor. The discussion contains important diagnostic and therapeutic problems.

[Modern therapy of cardiac insufficiency]. / Savremena terapija srcane insuficijencije.

UNLABELLED: Heart failure is a lethal, end-stage cardiovascular disease. Recent decrease in mortality rates from cardiovascular diseases has not been accompanied by a reduced mortality from heart failure. Survival, once the heart has used up all its reserves and compensatory mechanisms, is a little better than in cancer. That makes heart failure one of the most important world health problems. MATERIAL AND METHODS: This paper briefly reviews history, present and future of heart failure therapy, as a worldwide problem. Definition and diagnosis of heart failure, mechanism of deterioration of heart function, clinical use of present drugs in heart failure therapy and need for prevention of heart failure are, briefly pointed out. Results of clinical studies are presented, as well as the recommended indication for drug use. Heart failure is not readily identified, defined and evaluated. There is an absence of clear definition. To find a new definition which complies best with clinical practice is an important challenge cardiologists must face. Heart failure is a progressive disease and once the process has started it continues with further deterioration of cardiac function or ends in sudden death. In many patients changes within heart develop long before clinical symptoms occur. The left ventricle goes through a number of adaptations remodeling to compensate increased pressure or volume load or subsequent myocardial infarction. RESULTS AND DISCUSSION: For decades therapy has been focused on relieving symptoms, whereas preventive aspects and prolonging survival received less attention. Conventional therapy with diuretics and cardiotonic glycosides causes regression of symptoms and signs of heart failure, but there is no evidence that these drugs slow down the progression of the disease and reduce mortality. Currently, angiotensin-converting enzyme inhibitors plus diuretics are considered first line therapy for all degrees of heart failure, except for heart failure with atrial fibrillation and a rapid ventricular rate. They are, currently, the only agents with proven ability to decrease mortality. There is evidence about the efficacy of angiotensin-converting enzyme inhibitors in asymptomatic left ventricular dysfunction. It is likely that not only patients with significant reduction of systolic function but also other signs of impaired left ventricular dysfunction will benefit from treatment with ACE inhibitors. However, only preventive treatment may decrease the number of patients with new onset of clinical heart failure. Therefore treatment should be introduced early, rather than waiting for heart failure to progress to a more severe stage. Based on these facts ACE inhibitors should be considered as treatment of choice, a first line therapy in most cases of heart failure. A substantial reduction in cardiovascular mortality requires detection and correction of presymptomatic left ventricular dysfunction and risk factors, which predispose to its occurrence. Major contributors to the development of cardiac failure have been delineated and quantified. Identification of high-risk individuals is difficult since signs and symptoms of heart failure are often lacking. A strategy to find these patients must use objective methods to characterize the state of the left ventricle. Despite this, methods for efficient identification presymptomatic candidates for cardiac failure for preventive measures have been developed. High-risk candidates can now be costly-effectively targeted for treatment to delay failure. Early diagnosis and subsequent aggressive medical or surgical treatment are therefore fundamental to improve adverse outcome of heart failure. It is necessary to allow a rational approach to the clinician, indicating the effectiveness of these drugs in patients with evidence of impaired ventricular function. CONCLUSION: Further reduction of morbidity and mortality from heart failure and diseases associated with heart failure is to be expected by early

[Current knowledge of cardiomyopathy]. / Savremena shvatanja kardiomiopatije.

In 1961, 1964 and 1970, Goodwin proposed a somewhat unwield definition of cardiomyopathies, which was subsequently shortened to "Heart muscle disease of unknown cause" and separated from heart muscle diseases due to conditions involving organs other than the heart: the "specific heart muscle diseases". The cardiomyopathies thus defined were divided into hypertrophic, dilated and restrictive types and their classification was confirmed by the WHO/ISFC Task Force and the report of the WHO Expert Committee on cardiomyopathies in 1984. The cardiomyopathies are now classified by the dominant pathophysiology or where possible by aetiological/pathogenic factors. The modifications proposed by Dr Peter Richardson are timely and reflect revised ways of looking at problems that still face us. They are classified as dilated cardiomyopathy, hypertrophic cardiomyopathy, restrictive cardiomyopathy, and arrhythmogenic right ventricular cardiomyopathy. Unclassified cardiomyopathies include a few cases that do not fit readily with any group. The term specific cardiomyopathies is now used to describe heart muscle diseases which are associated with specific cardiac or systemic disorders.

[Relation between mitral orifice surface area and extent of hemodynamic disorder of the pulmonary circulation in patients with mitral stenosis]. / Odnos izmedu povrsine mitralnog usca i stepena hemodinamickih poremecaja u plucnoj cirkulaciji kod bolesnika s mitralnom stenozom.

UNLABELLED: Mitral annulus and valves form the mitral orifice area with the size between 4.0-6.0 cm2. Every area which is smaller than this, represents mitral stenosis. As a consequence of mitral stenosis hemodynamic gradients occur over the mitral orifice with circulation disturbances below and above the stenotic mitral valve. The size of transmitral gradient is important in the evaluation of functional or/and structural changes in the blood vessels of pulmonary circulation. This investigation included 40 patients with mitral stenosis (or accompanying minimal mitral regurgitation). All patients underwent echocardiographic examination: area of the mitral orifice was determined and hemodynamic procedure with the left and right heart catheterization was performed. The following hemodynamic parameters were measured: mean capillary wedge pressure, left ventricular filling pressure, left ventricular mean diastolic pressure, mean pulmonary artery pressure. According to these parameters resistance in the pulmonary circulation was measured. The size of the mitral orifice was determined according to oximetry blood analyses and hemodynamic parameters. All patients were divided into 4 groups: minimal (2.5-4.0 cm2), mild (1.5-2.5 cm2), moderate (1.0-1.5 cm2) and severe mitral stenosis (1.0 cm2). The comparison of echocardiographic and hemodynamic parameters revealed a high and positive correlation between the area of mitral orifice. There was also a negative and moderate correlation between the values of stenotic mitral orifice area and total pulmonary resistance, i.e. in all patients with severe mitral stenosis there was an increased pulmonary arteriolar resistance. CONCLUSION: Noninvasive echocardiographic method is valid in the evaluation of stenotic mitral valve area. In the evaluation of hemodynamic parameters in the pulmonary circulation the index of arteriolar pulmonary systemic vascular resistance is very important. In all patients with the area of stenotic mitral orifice 1.0 cm2, there are functional or pathomorphologic changes in the pulmonary circulation of the blood vessel wall.

[Evaluation of ejection fraction and end-diastolic volume using one-dimensional and two-dimensional echocardiography]. / Procena istisne frakcije i enddijastolnog volumena pomocu jednodimenzionalne i dvodimenzionalne ehokardiografije.

A group of 30 healthy persons and 120 coronary patients were investigated by one and two-dimensional echocardiography and digital subtraction angiography. End-diastolic volume and ejection fraction were estimated. Patients with coronary disease had greater volumes than healthy persons and smaller ejection fraction as well. The correlation of the given values showed that data gathered by two-dimensional echocardiography have greater coefficient in comparison to digital subtraction angiography (end-diastolic volume index r = 0.86, ejectional fraction r = 0.88, p < 0.01) than the same data estimated by one-dimensional technique. Two-dimensional echocardiography is a reliable noninvasive method for evaluation of these significant indicators of left ventricle function.

[Hypertension as a risk factor in acute infarct in young people]. / Hipertenzija kao rizik u akutnom infarktu mladih.

INTRODUCTION: Hypertension is one of the most frequent risk factors for acute myocardial infarction. The aim of this paper was to establish both the incidence of hypertension as a risk factor for myocardial infarction in young people and its characteristics. METHODS: The investigation involved patients younger than 40 years of age with acute myocardial infarction treated at the Institute of Cardiovascular Diseases in Sremska Kamenica from 1989 to 1994. RESULTS: The investigation involved 80 patients with acute myocardial infarction younger than 40 years--40 male and 40 female. Hypertension as a risk factor was found in 22 (55%) women and in 14 (35%) men and it was a moderate hypertension of systolic-diastolic type or unstable hypertension. In 10 (27.77%) investigated patients hypertension was detected during hospitalization (before that, it had not been controlled) while in 22 (61%) it was not seriously taken into account either by the patients, or their physicians and therefore it was not treated. As a single risk factor hypertension was found in 4 (10%) women, while it was not found in men. DISCUSSION: Hypertension is found in 10-15% of population, and in 10% young men and women. The existence of hypertension as the only risk factor for the occurrence of acute myocardial infarction was sufficient in these women, although it was considered that women in their generative period are protected from ishaemic heart diseases by their hormonal status. All that points to hypertension as a secret and mysterious killer. CONCLUSION: Hypertension as a risk factor was found in 22 (55%) women, and in 14 (35%) men. In 10% of women hypertension was the only risk factor, while it was not found as such in men. It was quite sufficient in these women for the occurrence of acute myocardial infarction, although it was considered that women in their generative period are protected from ishaemia in heart diseases by their hormonal status. Detection and adequate treatment of hypertension are of great importance as well as elimination of other risk factors.

Level of left ventricular dysfunction in endomyocardial biopsy-proven myocarditis.

The level of myocardial functional impairment in the course of myocarditis still remains associated with controversial data. However, many investigators agree that there is a significant heart failure during myocarditis and a large number of studies suggest development of global chronic or acute heart failure. The objective of this study was to establish haemodynamic parameters as descriptors of the level of myocardial systolic and diastolic impairment. From 131 investigated patients, 95 were assigned to the study group and 36 to the control group. All patients underwent right and left heart catheterization. A group of patients underwent EMB (endomyocardial biopsy) of the right ventricle. According to the Dallas Criteria, patients were divided into three groups: active myocarditis with fibrosis, active myocarditis without fibrosis and borderline myocarditis. The following haemodynamic parameters were evaluated: left ventricle EF (ejection fraction), cardiac index and end diastolic pressure of the left ventricle. EF, which reflects left ventricular systolic impairment, showed a difference related to the control group, while there was no difference in cardiac index between the groups. Diastolic dysfunction (end diastolic pressure of the left ventricle) was noted in the group with active myocarditis, but not in the group with borderline myocarditis.

The incidence of myocarditis in endomyocardial biopsy samples from patients with congestive heart failure.

We present the combined experience of three Yugoslavian cardiovascular centers in the application of endomyocardial biopsy for the diagnosis of myocarditis in patients who present clinically with congestive heart failure. The study group comprised 107 patients (mean age, 40.8 years; range, 19 to 61 years). On the basis of patient history and diagnostic tests, the following clinical diagnoses were established: dilated cardiomyopathy (85), myocarditis (16), and alcohol-induced heart disease (6). EMB samples were taken from the left ventricle (95) or both ventricles (12) by use of a King's College bioptome, with a mean of 3.2 samples per patient. Histologic evidence of myocarditis was noted in 10 of 85 patients (12%) with a clinical diagnosis of dilated cardiomyopathy, in 2 of 6 patients (33%) with alcohol-induced heart disease, and in 12 of 16 patients (75%) with a clinical diagnosis of myocarditis. There was confirmation of the clinically suspected diagnosis in 63% of cases, a change of diagnosis based on histology in 15% of cases, and nonspecific findings in 22%. However, useful information was obtained in 78% of the cases, and there was a 22% incidence of histologically proven myocarditis for the entire group. Our results indicate that endomyocardial biopsy is beneficial in determining the true incidence of myocarditis in patients with a clinical presentation of dilated cardiomyopathy.

Ultrastructural aspect of myocarditis: its relevance for the diagnosis.

The Dallas consensus was used to reveal active or borderline inflammatory loci by light microscopy (LM). When lymphocyte-cardiocyte interaction was observed by electron microscopy (EM), the deleterious or dormant pattern of inflammatory process was recognized. The first was determined by lymphocytes that adhered to cardiocytes, next to necrotic cardiocytes or admixed with debris. The second was marked by scattered lymphocytes between preserved cardiocytes and the absence of lymphocytes adhered to cardiocytes and necrotic cardiocytes. The deleterious pattern of the inflammatory process (EM) commonly supplemented the active appearance of inflammatory loci (LM). In contrast, the borderline outlook of the LM completed either the deleterious or dormant pattern of the EM. This discrepancy was related to the restricted resolution of LM, which might hide the actual stage of the disease. The diagnosis of myocarditis was founded on mutual LM and EM observations. The active or borderline appearance of LM of the deleterious pattern (EM) was considered indicative for the active stage of myocarditis. The borderline outlook of the LM of the dormant pattern of the EM was admitted to indicate either the healing phase of the disease with lymphocytes still lagging behind, or a latent phase of the ongoing myocarditis, according to the patient's hemodynamic status.

Autopsy material analysis at the Cardiology Clinic in 1999.

Clinico-pathological studies serve as a valuable source of information in everyday practice of most medical institutions. The aim of this study was to correlate the clinical and pathological diagnoses of principal disease and cause of death after autopsy in patients who died during 1999 at the Cardiology Clinic, Institute of Cardiovascular Diseases, Sremska Kamenica, Yugoslavia. Medical histories and autopsy reports of dead patients were analyzed: 255 (8.46%) patients died, but only 72 (28%) underwent autopsy. In 53 patients (73%) clinical cause of death was cardiac. Among these patients, consequences of coronary (atherosclerotic) artery disease prevailed in 41 patients (77%). Noncardiac cause of death was established in 19 patients (27%) and vascular causes prevailed with 17 (23%). The comparison of clinical and principal disease established by autopsy revealed a complete concordance in 62 cases, partial concordance in 1 and no concordance in 9 cases. Thus, concordance was found in 86-87.5%. The correlation coefficient was 0.48. Comparison of clinical and autopsy diagnoses of causes of death revealed concordance in 53 patients, with correlation coefficient 0.26.

[Coronary score in patients with low ejection fraction after myocardial infarct]. / Koronarni skor u bolesnika sa niskom ejekcionom frakcijom nakon prelezanog infarkta miokarda.

INTRODUCTION: The most frequent cause of heart failure is ischemic heart disease (1). This paper was aimed at comparing the coronary score of patients with low ejection fraction whose ejection fraction was not significantly changed after sustained myocardial infarction. MATERIAL AND METHODS: The study involved patients after sustained myocardial infarction treated at the Institute of Cardiovascular Diseases in Sremska Kamenica. Total coronary score and score of each individual coronary artery were emphasized. RESULTS: The investigation study comprised 56 patients aged 33-83 years of various occupations. Patients were divided into two groups: the first--A group consisted of 28 (50%) patients with ejection fraction 35% or lower; the second--B group also consisted of 28 (50%) patients with ejection fraction higher that 35%. Table 1. shows the dominant coronary artery in investigated groups of patients. Table 2. shows values of total and scores of each coronary artery. The right coronary artery was dominant in 75% of patients from the A group and in 82.1% of patients from the B group. A significantly higher individual score of coronary arteries, as well as the total score, was established in the group of patients with low ejection fraction, and especially the score of the anterior descendent artery which is almost twice higher in regard to the second group of examined patients. Table 3. describes the analyzed score in male and female patients. DISCUSSION: Patients with low ejection fraction after sustained myocardial infarction have more changes of coronary arteries than patients with better ejection fraction. The total score, score of the right coronary artery (ACD), circumflex artery (RCX) and especially anterior descendent artery (LAD) are significantly higher in patients with ejection fraction lower than 35%. There are no differences in the dominant coronary artery in investigated patients. In both investigated groups women had a smaller score of ACD and RCX and a higher score of LAD, but the difference is not significant. In regard to total score there were no differences in men and women. Numerous investigations also point to the fact that patients with lower ejection fraction and ischemic heart disease have more changes on coronary artery than patients with better ejection fraction. CONCLUSION: 1. Patients with low ejection fraction after sustained myocardial infarction have a higher total score and scores of ACD. LAD and RCX. 2. There are no differences in coronary score of men and women within the same investigated groups. 3. There are no differences in dominant coronary artery in investigated groups of patients.