Search

The aim of this research study was firstly to investigate the implications of the psycho-social aspects of haemophiliacs/carriers in Greece and secondly to understand human nature and thus be able to handle haemophiliacs' and carriers' anxieties and fears. The design and methodology of the study was an exploratory and descriptive one. The total membership was a random sample of hundred haemophiliacs and hundred carriers. Only 89 and 75 respectively answered. Two questionnaires were administered with a veiw to obtain the groups' reactions and attitudes to various psycho-social aspects affecting their personal life and environment. The questionnaires consisted of open-ended and closed questions. The findings from this study suggested that haemophiliacs experience pain, parental anxiety, stress in the family and interference with family plans in a more realistic and objective way. This attitude could be a result either from a genuine awareness of their disease or from the psychology of defending oneself through denial. With regard to carriers it was found that similar psycho-social aspects had significant signs of exaggeration in terms of fear and anxiety. This could be attributed to the inappropriateness of their defensive reaction. The evaluation of the responses in the light of the actual situation, confirms the above results with an even greater emphasis on tendencies.

Sixty unrelated Greek patients with haemophilia (46 with haemophilia A and 14 with haemophilia B) were typed for HLA-A, B and DR antigens. A highly significant increase in the frequency of HLA-DR5 was observed in both groups of patients (58.6% vs 30.0%, chi 2 = 10.47, pc less than 0.03, RR = 3.31 for haemophilia A and 78.5% vs 30.0%, chi 2 = 12.32, pc less than 0.007, RR = 8.5 for haemophilia B). An increased frequency of HLA-B13 was also observed in patients with haemophilia A (15.2% vs 5.7%, chi 2 = 5.74, pc less than 0.4, RR = 2.9). Thirty of the 60 patients (50.0%) were positive for LAV/HTLVIII antibodies. HLA-DR5 was equally distributed in patients with and without these antibodies (63.3% and 63.3%, respectively). The presence of DR5 did not correlate with the severity of haemophilia A or B. These results may suggest an influence of gene(s) on chromosome 6 in haemophilia A and haemophilia B and no effect of HLA antigens in the susceptibility to LAV/ HTLVIII infection among haemophiliac patients.

Our objective was to assess the influence of genetic factors such as HLA classes I and II antigens and other clinical and laboratory variables on the progression of HIV disease in a cohort of 118 HIV infected haemophilic subjects of Greek origin who had been typed for HLA antigens and were followed up prospectively for 22 years since seroconversion. At the end of the follow up we compared two groups of patients: 22 patients who had a fast progression to AIDS (median 6 years since seroconversion) vs. 33 patients who remained asymptomatic in stage A2 for up to 22 years (median 15 years). The results showed that the two groups did not differ significantly in age at seroconversion or baseline CD4+ T cell count. However there was a difference in the frequencies of certain HLA antigens in the two groups. The fast progressors had a higher frequency of HLA-A28, B21 and DR3, which was statistically significant (P = 0.02, 0.04, 0.05, respectively) compared to the slow progressors. These findings based on classical HLA typing techniques confirm other published observations and support the effect of genetic background in the progression of HIV infection in haemophilics.

Our objectives are to describe the progression of HIV disease and to assess the influence of hemophilia-related variables, age at infection, and antibodies to cytomegalovirus infection (anti-CMV) in a Greek cohort of 158 HIV-1-positive hemophilic men, who received prospective follow-up for up to 16 years after infection. A total of 79 patients had died, representing a cumulative progression rate of 72.4% (95% confidence interval [CI], 56.6-83.3). A significant proportion of the mortality (30%) resulted from conditions not formally related to AIDS, with liver failure and cerebral hemorrhage predominant. At 16 years after seroconversion, 66 patients had developed clinical AIDS, a cumulative progression rate of 58.2% (95% CI, 47.1%-86.3%) whereas 15 years after infection 81.5% (95% CI, 74.2%-87.9%) of the patients had AIDS or a CD4 cell count200 cells/mm3. Hemophilia-related variables or presence of anti-CMV were not significantly associated with disease progression. Age at infection was a strong prognostic factor for all three endpoints. Appropriate modeling showed a nonlinear age effect, with a steeper increase of relative hazard for patients40 years of age at seroconversion. The age effect remained significant even after controlling for current CD4 cell count. Further investigation is required to elucidate the mechanisms of the age effect and the contribution of HCV coinfection on the disease progression.The Greek Hemophilia Cohort Study encompasses 158 HIV-positive men with documented seroconversion dates. The present study estimated the rate of progression to all-cause mortality, clinical AIDS, or advanced immunodeficiency 16 years after seroconversion and evaluated the independent effects of hemophilia-related factors, age at seroconversion, and cytomegalovirus status early in the course of infection. Seroconversion dates extended from September 1980 to December 1985. By 1996, after a median follow-up of 11.6 years, 117 of the 158 men had developed AIDS or had a CD4 cell count of 20 cells/cu. mm, and 79 had died. The estimated cumulative incidence rates of clinical AIDS and death over 16 years since infection were 58.1% and 72%, respectively. 30% of the mortality was due to diseases not formally associated with AIDS (e.g., liver failure and cerebral hemorrhage). A significant association existed between older age at seroconversion and more rapid progression to both AIDS and death, with a particularly steep gradient for patients over 40 years old at seroconversion. The age effect remained significant even after controlling for current CD4 cell count. Increases of 1.32-fold and 1.33-fold in the risks of dying and developing AIDS, respectively, were obtained with every one unit decrease in CD4 cell count on the square root scale. Severity of hemophilia, dosage of clotting factor concentrates, and antibodies to cytomegalovirus were not associated with either AIDS risk or mortality. Further investigations are urged to clarify the mechanisms underlying the age effect observed in this study.

Summary. We illustrate the usefulness of direct mutation detection for genetic counselling by showing its application to an extremely large mild haemophilia A pedigree (91 haemophiliacs) originating from the village of Aiani in Macedonia, northern Greece. The causative mutation has already been shown to be an A to T transversion in codon 280 of the FVIII gene which replaces Asn 280 (AAC) by He (ATC) and which creates a new Bam HI restriction site in exon 7. The latter permitted direct, rapid and reliable detection of the mutation in relevant family members. All major branches of the family were shown to share the mutation, and carrier status was diagnosed or excluded for 23 possible carriers. Other interesting characteristics of the Aiani haemophilic population are a slightly higher longevity and fecundity than that observed in the general population and a wide range of FVIILC levels (5–25%) associated with the mutation.

Previous data indicate that even mild postprandial hyperglycaemia in diabetic subjects, who are concerned to be in good control, activates haemostasis. The aim of this study was to investigate the effect of the oral administration of 5 mg glibenclamide on postprandial activation of coagulation in type 2 diabetics.We designed a placebo controlled, randomised study. After an overnight fast, each subject (n = 16, age 50-68 yr.) underwent a standard test meal (600 Kcal: carbohydrates 40%, lipids 50%, proteins 10%) preceded by one tablet of glibenclamide (5 mg) or placebo. The two tests were performed randomly, with an interval of 7 days. Blood samples were collected at baseline and 2 and 4 hours after the meal to measure the concentrations of glucose, insulin, c-peptide, triglycerides as well as of d-dimers, fibrinogen, F1.2 and TAT. The postprandial levels of TAT, fibrinogen, F1.2, d-dimers, insulin, glucose and triglycerides were significantly higher compared to baseline values.The postprandial levels of glucose, triglycerides, fibrinogen, F1.2, TAT and d-dimers were lower after glibenclamide administration as compared to placebo, while the concentrations of insulin and c-peptide were higher. Thus, acute administration of glibenclamide reduces the postprandial activation of coagulation.

To detect and characterize parvovirus B19 infection during the course of progressive immune deficiency from human immunodeficiency virus (HIV), ten subjects enrolled in the Multicenter Hemophilia Cohort Study were followed for 6.4 to 15 years from HIV seroconversion through extreme immune deficiency. Four to five sera or plasma samples from each subject, collected at predetermined CD4+ lymphocyte levels, were tested for immunoglobulin G (IgG) and M (IgM) B19 antibodies and DNA. All 42 samples were positive for B19 IgG antibodies, and three were weakly positive for IgM antibodies. Only one sample, collected coincident with HIV seroconversion, was unequivocally positive for B19 DNA. No persistent hematologic adverse effects of B19 infection were observed. Thus, although B19 IgG antibodies are highly prevalent among HIV-infected persons with hemophilia or related disorders, B19 viremia and its hematologic consequences were not detected, even with severe depletion of CD4+ lymphocytes. If primary B19 infection occurs after immune deficiency, however, the consequences may be more adverse.

The frequencies of HLA antigens in 33 HIV seronegative and in 88 HIV seropositive haemophiliacs, who have been followed for at least 6 years since seroconversion or first HIV positive test. were evaluated in relation to disease susceptibility and disease progression. A high frequency of HLA-A2 and -DR2 antigens and a low frequency of HLA-A9 were found to characterize HIV seropositive patients (p

The antibody responses and the prevalence patterns of antibodies to hepatitis C virus (anti-HCV) in a cohort of patients (n = 210) with bleeding disorders were studied using a first-generation and a second-generation enzyme immunoassays (EIA-1, EIA-2) as well as a second-generation recombinant immunoblot assay (RIBA-2). The anti-HCV prevalence as determined by EIA-1 and EIA-2 was 183/210 (87.1%) and 197/210 (93.8%), respectively (p = 0.0026). None of the 17 EIA-2(+)/EIA-1(-) samples was scored nonreactive by RIBA-2. At follow-up, samples of 123 patients were tested. Twenty-nine out of 111 patients reactive by EIA-1 seroreverted according to EIA-1 while the seroreversion rate with EIA-2 was 0 out of the 121 (p10(-8)). The anti-HCV prevalence by EIA-2 was 150/154 (97.4%) in anti-HIV-1-positive individuals and 47/56 (83.9%) in the anti-HIV-1-negative ones (p = 0.001). However, high assay signals (OD 492 nm2.0) were observed in 94/150 (62.7%) and 45/47 (95.7%) of the anti-HIV-1-positive and -negative patients, respectively (p = 10(-5)). The decreasing anti-HCV reactivity among anti-HIV-1-positive individuals was mainly due to diminishing c33c reactivity. Seroconversion to anti-HCV was observed in 3/7 (42.9%) cases with acute icteric non-A, non-B hepatitis by both EIA-1 and EIA-2, while the remaining 4 cases had detectable levels of anti-HCV 1-18 months before the acute episode.

SummaryPlatelet count and aggregation were assessed in 9 Finnish amateur runners aged 34 to 48, and one 65-year old taking part in a non-competitive marathon race (42.2 km).After the run the mean value of platelet count showed a very significant rise (p The finding of platelet hyperaggregability after prolonged strenuous exercise even in trained subjects is discussed. It is concluded that a thorough medical examination of the haemostatic balance is recommended before a marathon race.

Hemophilia B Leyden is characterized by low levels of factor IX antigen and activity before the age of 15 years, whereas after puberty factor IX levels rise at a rate of about 5% per year. Two distinct point mutations (deletion of A, A----G) were identified at position +13 of the factor IX gene of a Greek and an American patient with hemophilia B Leyden. The nucleotide changes have occurred 32 basepairs downstream of a previously reported point mutation in a Dutch kindred with the same hemophilic phenotype. The results point to the importance of sequences surrounding the putative start site for the constitutive expression of the factor IX gene and to the possible significance of an imperfect direct repeat of DNA.

SummaryA short review is given on several aspects of the management of haemophilia in Greece. A total number of 429 patients with congenital haemorrhagic disorders has been registered. The incidence of F. VIII deficiency is 85.9% and that of F. IX 12.9%. Information concerning distribution throughout the country, age, social standard, financial position and education is given too.Haemophiliacs are systematically treated by Athens specialized Haemophilia Treatment Centres.The most widely therapeutic material still used, is fresh frozen plasma. Lyophilized cryo-precipitate and F. IX (prothrombincomplex) concentrate are also prepared but they are not for every day use. Imported concentrates are used only for very serious cases.Home therapy is attempted in limited scale and it is largely discussed.Consultation, out-patients treatment and hospitalization, if needed, are free of charge for haemophiliacs regardless of their financial position or their national insurance.

SummaryBlood coagulation and fibrinolysis were assessed in 13 Finnish amateur runners aged 31 to 48, and one 65-year old taking part in a non-competitive marathon (42.2 km). After the run the mean values of partial thromboplastin time showed a very significant shortening, whereas the mean values of the prothrombin time and of plasma fibrinogen were not significantly altered. The mean values of euglobulin lysis time were significantly shorter and the mean values of fibrin degradation products increased highly significantly. After the run, protamine sulphate was positive or strongly positive in all subjects, whereas the ethanol gelation test was negative in all runners; no cryofibrinogen was detected in any participant. Thus, running a marathon race affects the haemostatic balance and activates the fibrinolytic mechanism. The effects of training and physical fitness on the above parameters are discussed.

SummaryBlood coagulation, fibrinolysis, platelet count-aggregation and Cortisol were assessed in 35 Finnish amateur runners aged 27 to 56 years (mean 40) and three aged 65, 67, and 82 who had run a non-competitive Marathon in 1975, 1976 and 1977 over the classical itinerary. After the run, in all 3 years, APTT showed shortening (p < 0.001); prothrombin time and plasma fibrinogen were not significantly altered; euglobulin lysis time was shorter (p < 0.001) and FDP increased (p < 0.001); PSPT became positive in all subjects, whereas the ethanol gelation test remained negative; no cryofibrinogen was detected. Platelet count and aggregation showed increase (p < 0.001) in 1975 (extreme heat, 25° C) but remained unaltered in 1976, 1977 (15–18° C). Cortisol levels were always significantly increased – more markedly in 1975. Women’s responses were similar to those of men. A possible correlation between physical fitness and the responses of haemostatic balance is suggested and the influence of weather is discussed.

Peripheral blood lymphocyte subsets and the incidence of LAV/HTLV-III antibodies were studied in 63 patients with hemophilia A who had been transfused with a low dose regimen of commercial (U.S.A.) factor VIII concentrates. Five patients with hemophilia B were also included in this study. In hemophilia A patients a significant reduction in the percentage and absolute numbers of T4+ cells and of the T4/T8 ratio and a significant increase in the percentage of T8+ and Leu-7+ cells were observed. These abnormalities were independent of the presence of anti-LAV/HTLV-III. In hemophilia B patients a significant increase of T8+ cells and a decrease of T4/T8 ratio was noted while the percentage of Leu-7+ cells was normal. A significant negative correlation of F VIII units transfused and T4/T8 ratio was seen only in LAV/HTLV (-) patients, suggesting that F VIII per se could cause immunodysregulation. Seropositive patients were found to have consumed a larger amount of F VIII units than seronegative patients during the period 1980-1984 (p less than 0.005).

SummaryBlood coagulation, fibrinolysis and platelet aggregability were assessed in 8 physicians aged 30-40 years, who had travelled nonstop by car from Salonica to Athens (510 km) and returned to Salonica after 48 h of rest and after administration of 1 g of aspirin. At the end of journey A, platelet aggregability was found to be increased (6 out of 8 persons), AT III was decreased by 30% (p A common finding of both journeys was the increase of platelet counts at the end (p The correlation between long lasting sitting and the response of the haemostatic balance is suggested. The influence of aspirin is discussed.

Venous thrombosis in childhood is an extremely rare condition. The etiologic factors in childhood are different than in the adult. Generalized infection seems to play the main etiologic role, and for this reason antibiotic treatment is usually necessary. Anticoagulant treatment is mandatory; however its application presents many difficulties. The administration of calcium-heparin subcutaneously for a long time is feasible, well-controlled, without side effects and highly effective. The authors therefore strongly recommended this anticoagulant treatment in childhood.

A study of various haemostasis parameters in a group of twenty-two traffic policemen working under special conditions, i.e. in locations of heavy traffic (5 hours shifts in the centre of Athens) is presented. These normal subjects (aged 22-39) are subjected to physical and psychological stress and to the effect of increased air pollution. Pretests were conducted before the beginning of the shift (6 a.m.) and postests were conducted at the end of the shift (12 noon).The following tests were performed: 1. Prothrombin Time (P.T.), 2. Activated Partial Thromboplastin Time (A.P.T.T.), 3. Fibrinogen, 4. Euglobulin Lysis Time (E.L.T.), 5. Fibrin Degradation Products (F.D.P.), 6. Platelets, 7. Platelet Aggregation by A.D.P., 8. PF4 9. F VIII:C, F VIII:Ag, F VIII:Rcof, 10. Antithrombin III (AT III).Results: Significant decrease of Fibrinogen, ELT and AT III, significant increase in APTT, FDP. Increase in PF4 levels. The results are indicative of the occurence of an activation of the fibrinolytic mechanism after the shift with signs of consumption of certain coagulation factors. There is an adverse effect on the hemostatic balance of traffic policemen during their service.

Serum samples from 134 Greek haemophiliacs multitransfused with fresh frozen plasma and factor - VIII or factor - IX concentrates were investigated for the presence of blood group antibodies. All samples were tested against a panel of selected cells of known but differing genotype. The incidence of blood group antibodies was 12.7 96. Anti-D was found in 6 out of the 17 immunized patients, i.e., 35.3 % of D-negative haemophiliacs were sensitized. It is concluded that the high incidence of immunization is a serious complication of repeated plasma transfusions.

About 250 individuals belonging to 44 families with hemophilia A or B were studied in our laboratory. The detection of carriers was first established by pedigree analysis of each family . and coagulation and immunological assays of factor VIII or IX. The availability of specific probes for the molecular study of these two genes makes possible a diagnosis with certainty in the case of informative families. 25 families of hemophilia A were studied. For each person, blood was collected into EDTA and leucocyte DNA was extracted, digested by restriction endonucleases, electrophoresed in 0.9 % agarose gels and transferred to nitrocellulose filters by Southern blotting. Two probes were used for the analysis of factor VIII gene. The St 14 probe (J.L. Mandel) located on the q28 region of the X chromosome and closely linked to the gene, determines a restriction fragment length polymorphism (RFLP) when the DNA is digested by the enzyme TaqI. The p114-12 genomic probe (Genentech) corresponding to the exons 17 and 18 of the factor VIII gene, reveals a RFLP in the DNA digested by the enzyme BclI. 19 families -of hemophilia B were studied. A total factor IX cDNA probe was used for the screening of potential deletions in the case of hemophiliacs with circulating antibodies. A genomic probe containing the exons II, III and IV of factor IX was used to detect the TaqI RFLP. For the study of factor VIII gene, the extragenic probe St 14 gives a very high percentage of informativity (about 90 %) but recombination can occur between the probe and the gene. The p 114-12 probe, which is used to confirm the results given by the St 14 probe, gives about 20 % informativity. In our study, we were able to diagnose carrier state with certainty in 92 % of the families. For hemophilia B, the genomic probe gives about 40 % informativity. A large deletion of the region of the factor IX gene has been found in one family and remains to be mapped. In conclusion, carrier detection and prenatal diagnosis can be established with certainty by molecular studies in most cases of hemophilia A using the St 14 probe, with a 5 % risk of recombination when the BclI RFLP cannot confirm. This diagnosis is possible in about 40 % of the cases of hemophilia B.

A number of joints suffering from haemophilic arthropathy was studied. They were surveyed clinically, radiologically, and with a new system of whole-body gamma-camera scintigraphy. This radio-isotopic technique provides advantages such as sensitivity, speed and safety. The results of the survey are presented and discussed.

Blood coagulation, fibrinolysis and platelet aggregability were assessed in 8 physicians aged 30-40 years, who had travelled non-stop by car from Salonica to Athens (510 km) and returned to Salonica after 48 h of rest and after administration of 1 g of aspirin. At the end of journey A, platelet aggregability was found to be increased (6 out of 8 persons), AT III was decreased by 30% (p less than 0.001), the F VIII:C / F VIIIR: Ag ratio was decreased (p less than 0.02) and ELT was prolonged. At the end of journey B the findings were the following: platelet aggregation was not affected, the decrease of AT III was not statistically significant and ELT was significantly shortened (p less than 0.005). A common finding of both journeys was the increase of platelet counts at the end (p less than 0.005). The correlation between long lasting sitting and the response of the haemostatic balance is suggested. The influence of aspirin is discussed.

Two hemophiliac patients with nephrolithiasis were treated with extracorporeal lithotripsy. Both patients were followed up by the 2nd Hemophilia Center of Athens. One is registered as severe hemophilia A and the other one suffers from hemophilia B. With the cooperation of the specialized center they were prepared as for a major operation by administration of substitution therapy. The least possible number of shock waves at low voltage was administered in order to minimize renal damage. Both patients had an uneventful postoperative course.

In this paper, a five generation Greek family is described with haemophilia B. The disease is characterized by a normal ox-brain prothrombin time, normal levels of the vitamin-K dependent clotting factors VII and X and a proportional reduction of factor IX activity and antigen levels all of which is consistent with the cross-reacting material negative form of haemophilia B. However, in this family the factor IX levels in the three patients of generation V are around 1 U/dl while the three older patients in generation III have factor IX levels ranging from 28 to 44 U/dl. In the oldest patient of generation V we observed a rise of the factor IX level from 1 U/dl up to the age of 13 to 10 U/dl at age 14. In addition, the older patients have very mild bleeding symptoms or none at all, while the young ones have occasional spontaneous haemorrhages in muscles and joints, compatible with severe or moderately severe haemophilia. The disease appears to be similar to haemophilia B Leyden which has been described in a Dutch family.