Juraj Sepčić

Nutritional Factors and Multiple Sclerosis in Gorski Kotar, Croatia

An analysis was made of the possible influence of nutritional factors on the etiology of multiple sclerosis in Gorski Kotar (Croatia), a high-risk zone for this disease. A total of 46 MS patients and 92 controls, native-born residents of the area studied, participated in a case-control study. The questionnaire comprised 51 questions concerning dietary habits. An odds ratio (OR) estimate was obtained for all the factors which were more frequently found in the patients than in the controls. Large differences were found in the daily consumption of different quantities of full fat unskimmed milk (OR 21.7; chi 2 42.34; LL 7.12), potatoes with lard and fresh or smoked meat (OR 20.7; chi 2 15.52; LL 2.72), and new potatoes (OR 20.7; chi 2 15.52; LL 2.72). The consumption of unpasteurized milk, animal fat, smoked meat and potatoes are nutritional risk factors which could have an influence on the severity of primary demyelinization in a high-risk area for multiple sclerosis.

Mutations in the hemochromatosis gene (HFE) and multiple sclerosis

In the present study we have investigated whether HFE gene polymorphism may play a role in the disease process of Croatian and Slovenian MS patients and their potential genetic susceptibility to MS. We genotyped 314 MS patients and 400 healthy controls for the C282Y and H63D mutations by polymerase chain reaction/restriction fragment length polymorphism (PCR-RFLP) analysis. Our results showed no significant differences in the distribution of the two mutations between MS patients and controls, suggesting that HFE polymorphisms do not contribute to the susceptibility to MS. Also, there was no significant correlation between HFE polymorphism and the disease progression index. However, we observed that MS patients carrying the mutant C282Y allele exhibited earlier onset of disease symptom relative to other genotypes, but it warrants further study in a larger series of MS patients.

INCREASED PERFORIN EXPRESSION IN MULTIPLE SCLEROSIS PATIENTS DURING EXACERBATION OF DISEASE IN PERIPHERAL BLOOD LYMPHOCYTES

The expression of perforin (P) in subpopulations of the PBL of multiple sclerosis (MS) patients in stable and active phase of disease was investigated, by simultaneous detection of P (intracellular molecule) and cell surface antigens. A significant increase of CD4+P+ (p < 0.02) and CD16+P+ (p < 0.001), and decrease of CD56+P+ (p < 0.05) cells in active MS was found. In active disease there is a highly significant increase (p < 0.001) of average fluorescence intensity (AFI) for P in CD4(dim+) cells, and these cells are larger in size and have higher granularity (p < 0.05) compared to CD4(bright+) p(dim+) cells. Surprisingly, there were no CD25+P+ cells in either group of MS patients. These results show that CD4+P+ cells are upregulated in active disease in cell number, in the level of P expression per cell, and in the level of cell activation (increase in cell size and granularity). It is suggested that CD4+P+ cytotoxic cells may play a role in the pathogenetic mechanisms of MS.

Tumor Necrosis Factor-α-308 Gene Polymorphism in Croatian and Slovenian Multiple Sclerosis Patients

Previous findings regarding the role of TNF-α-308 gene polymorphism in multiple sclerosis (MS) are contradictory. The aim of this study was to investigate the possible influence of TNF-α-308 polymorphism on MS susceptibility and the MS disease process in a Croatian and Slovenian population. Genotyping was performed in 338 patients and 460 healthy controls. The TNF2 allele was present in 123 (26.8%) healthy controls vs. 67 (19.9%) MS patients (p = 0.023, odds ratio = 0.68, 95% confidence interval = 0.48–0.95), suggesting that carriage of the TNF2 allele might decrease MS risk. The difference in TNF2 allele carrier frequency between patients and controls was identified in the relapsing-remitting MS group. There was no association between TNF2 allele carrier status and age at disease onset or disease progression. Our results suggest that, in the study populations, the TNF-α-308 polymorphism may play a role in MS susceptibility.

Angiotensin-converting enzyme I/D gene polymorphism and risk of multiple sclerosis

Objectives –  Angiotensin‐converting enzyme (ACE) activity is increased in blood and cerebrospinal fluid of patients with multiple sclerosis (MS). In addition, in experimental autoimmune encephalomyelitis (EAE), an animal model of MS, the blockade of ACE suppresses the disease itself. To analyze the genetic association of the ACE gene with MS, we examined ACE gene insertion/deletion (I/D) polymorphism in MS patients.

No association of CCR5D32 gene mutation with multiple sclerosis in Croatian and Slovenian patients

Several studies investigating the role of the CCR5D32 mutation in multiple sclerosis (MS) have reported varied, often contradictory results. Therefore in the present study we have analysed whether the CCR5D32 mutation is associated with the risk of/or disease process in Croatian and Slovene MS patients. Three hundred and twenty-five MS patients and 356 healthy controls were genotyped by the polymerase chain reaction method. Our results showed no significant differences in the distribution of CCR5D32 mutations between MS and control subjects, indicating that this mutation does not influence susceptibility to MS. Furthermore, we did not observe that CCR5D32 carrier-status could modulate age of disease onset or progression of the disease. It is therefore our conclusion that the present study indicates that the CCR5D32 mutation is neither protective of, nor a risk factor, for MS development.

Epidemiology of multiple sclerosis in western Herzegovina

OBJECTIVES To determine epidemiological rates of multiple sclerosis (MS) in western Herzegovina. PATIENTS AND METHODS We analysed data from 81 MS patients (49 females, 32 males) on the prevalence day, 31 December 2003. Patient information was obtained from a search of all available medical records from the period 1994-2003 in the investigated area. RESULTS Crude prevalence of MS was 27/100,000 (95% confidence interval (CI) 20-34). Prevalence was highest in the mountainous municipality of Posusje (56/100,000) and lowest in the coastal municipality of Neum (0 incidence). The annual incidence of MS was 1.6/100,000 (95% CI 0-3.3). The female/male ratio of MS was 1.5. The mean age of the patients on prevalence day was 40.0+/-11.6 years, and the mean age at disease onset was 31.0+/-7.1 years. Eight (10%) of the patients had a first-degree relative with MS. The primary progressive (PP) disease course was observed only in females. Visual symptoms were the initial symptom of MS in 6 (7%) of the patients. CONCLUSIONS Western Herzegovina is an area of moderate risk for MS, and the distribution of MS in western Herzegovina is heterogeneous. PP-MS occurred only in females, and involvement of the visual pathways as the initial symptom of MS was low.

Epidemiology of multiple sclerosis in Croatia

In 1897 Franjo Gutschy reported two cases of multiple sclerosis (MS) in the Liecnicki Viestnik [1]. This was the first official report of pathologically verified MS in Croats [2]. Radoslav Lopasic noted that ‘MS is present particularly in a restricted area in the western parts of Croatia’ [3]. In 1969 one of us (JS) carried out the first systematic epidemiological survey of MS in that region [4]. The present paper is a compilation of the results of an analysis of the Croatian national medical bibliography from 1887 to December 2000. Its aim is to provide an exhaustive review of epidemiological descriptive, analytic and genetic observations on MS in Croatia [5 /9]. From 1897 to 1945, the few papers dealing with MS discussed the symptoms, signs and differential diagnosis of the disease, most of them focusing on the relation between MS and pregnancy [5,6] (Fig. 1). From 1946 to 2000, MS research intensified, covering mostly its laboratory aspects, immunology and epidemiology [7 / 9]. The major contributions to MS research in Croatia were made in the analysis of cerebrospinal fluid by the Zagreb neurobiochemistry group [10,11], in the role of Tand B-lymphocytes, and of natural killer cells, by the Zagreb and Rijeka immunology groups, [12 /15] and epidemiological research by the Croatian Reference Center for Demyelinating Diseases in Rijeka [16 /22] (Fig. 2). Neuroepidemiological and clinical studies of MS in Croatia reveal a high percentage of erroneous diagnoses. This suggests that physicians in general, as well as some neurologists, are not well acquainted with its differential diagnosis, that uniform diagnostic criteria and algorithms for the disease are lacking, that complementary examinations frequently are incomplete and that the diagnosis of MS seems to be made irresponsibly in order for the patient to obtain disability assistance (Table 1) [23] (L. Radolovic-Prenc, unpublished data, 2001).

Vaginally Applied Diquat Intoxication

CASE REPORT We report the case of a woman who introduced 20 mL of diquat concentrate intravaginally. Abdominal pain, vomiting, diarrhea, burning chest pains, and somnolence appeared within the first 24 hours. The vulva and vagina were corrosively inflamed. Acute renal failure appeared on the third day and was treated by 6 hemodialyses over 6 days. The patient was dysarthric with spastic tetraparesis for 3 months. The electroencephalogram, diffusely slow on day 3, was normal on day 28. The electromyoneurogram was normal at all times. Biopsy of a peripheral nerve performed on day 57 following intoxication showed no myelin or axonal alterations.

Interleukin 7 receptor alpha polymorphism rs6897932 and susceptibility to multiple sclerosis in the Western Balkans

The interleukin 7 receptor alpha single nucleotide polymorphism rs6897932 was identified as a multiple sclerosis susceptibility-modifying polymorphism in genome-wide and gene scan studies, mainly in populations in western countries. The aim of this study was to investigate the association of interleukin 7 receptor alpha rs6897932 with multiple sclerosis in populations from the Western Balkans: Serbia, Croatia, and Slovenia. A total of 678 unrelated white patients and 597 unrelated, ethnically matched healthy controls were included in the study. Genotyping was performed by real-time polymerase chain reaction. We found no significant difference in genotype or allele frequencies between controls and patients with multiple sclerosis either separately in Serbian, Croatian, and Slovenian populations or in the whole sample from the Western Balkans. The odds ratio for multiple sclerosis in this study was 1.04 (0.86—1.25) for the C allele. It is known that demographic as well as environmental factors have a substantial role in multiple sclerosis development, as well as population genetic background. The results of this study indicate that other types of genome variants should be required for the development and/or progression of multiple sclerosis, which may vary among populations.