Polyendocrine syndrome type 3C in a family from Pakistan.

In type 3 polyendocrine syndrome (PAS3), autoimmune thyroiditis occurs with other organ-specific autoimmune disease, but not with autoimmune adrenalitis. In this report we described a family from Pakistan in which mother and three daughters were affected by a PAS3. We studied a family from Pakistan: Father MMu age 44, mother KN aged 44, three daughters MM age 20, MH age 16 and MA age 14 and a son MU age 18. These subjects were tested for thyroids function, metabolic function, adrenal function, autoimmune disease. In this family the four females were shown hypothyroidism with presence of anti thyroid autoantibodies (AA) and high TSH serum concentration in association with the presence of anti transglutaminase AA. Moreover KN, MM and MH were positive for anti nuclear AA (granular pattern) and for antibodies against Saccaromyces cerevisiae. MM was positive for AA against nuclear extractable antigens (SSA and SSB) too. No diabetes or pernicious anemia were observed. Adrenal and Pituitary function were normal. PAS 3C is an uncommon disease. In this family from Pakistan we observed a PAS3C in the four female members: mother and three daughters while father and son were unaffected.

High prevalence of HFE gene mutations in hemodialysis patients.

AIM: Hemochromatosis (HH) was a common inherited disease characterized by iron overload. This disease is usually the result of mutations in the HLA-linked hemochromatosis gene (HFE). The aim of this study was to evaluate the frequency of HFE mutations in a group of Venetian hemodialysis patients. METHODS: Sixty-one hemodialysis patients, 62 patients with laboratory findings suggestive for iron overload, 57 repeat blood donors were enrolled in the study. HFE mutations were detected by using a commercial strip assay. RESULTS: In this study only H63D and C282Y mutations were observed. The overall prevalence of HFE mutations was 40.9% among hemodialysis patients, 30.6% among patients with laboratory findings of iron overload and 15.8% among blood donors. CONCLUSION: A high prevalence of HFE mutation among hemodialysis patients was observed. Prevalence of HFE mutation in this group was 40.9%, significantly higher than results observed among blood donors (15.8%, P<0.005) or among patients with laboratory signs of iron overload (30.6%, P<0.01). These data are, at present inexplicable, and this results need further confirmation.

TPMT genotype and the use of thiopurines in paediatric inflammatory bowel disease.

BACKGROUND: Thiopurines are used in the treatment of inflammatory bowel disease. They are metabolised via methylation by thiopurine-S-methyltransferase (TPMT), which displays a genetically determined polymorphic activity. Subjects with reduced TPMT activity have a higher concentration of active thiopurine metabolites and may be at increased risk of bone-marrow suppression. AIMS: To evaluate the relevance of TPMT genotyping in the management of thiopurines therapy in inflammatory bowel disease patients. PATIENTS AND METHODS: Adverse effects and clinical response were determined retrospectively and correlated with TPMT genotype in 70 paediatric inflammatory bowel disease patients. RESULTS: Nineteen patients (27.1%) developed adverse effects; of the 51 who did not, 34 (66.7%) responded to treatment. Five patients (7.1%) were heterozygous for a variant TPMT allele; two of these (40%) were intolerant to thiopurines, compared to 17 of the 65 patients (26.2%) with a wild type gene (O.R. 1.88, 95% CI 0.29-12.2, p=0.61); among the 34 responders, the median dosage of the drug required to obtain remission was lower for mutated than for wild type patients (1.6mgkg(-1)day(-1) versus 2.0mgkg(-1)day(-1), p=0.043). CONCLUSIONS: There was no significant association between adverse effects of thiopurines and TPMT heterozygous genotype, but TPMT genotyping could be useful in establishing the most appropriate dose of thiopurines to start treatment.

Biological qualification of blood units: considerations about the effects of sample's handling and storage on stability of nucleic acids.

BACKGROUND: In transfusional setting introduction of nucleic amplification technique (NAT) for HBV-DNA, HCV-RNA and HIV-RNA in biological qualification of blood units suggest some problems. At first the opportunity to operate on mini-pool, at second the need to store the samples at +4 degrees C. The authors therefore have tried to estimate the impact of these conditions on the operativity of NAT testing in the transfusional setting. METHODS: The following parameters has been estimated: distribution of viral-load in untreated subjects, stability of nucleic acids during storage at +4 degrees C, stability of nucleic acids after repeated cycles of freezing and defrosting, robustness of the test to the cross-contamination, definition of the detection-limit (95%). Quantitative tests has been performed by using the following kits: Cobas Amplicor HBV Monitor, Cobas Amplicor HCV Monitor, Cobas Amplicor HIV Monitor; the qualitative tests has been performed by using the following kits: Ampliscreen HBV, Ampliscreen HCV 2,0, Ampliscreen HIV 1,5 all supplied by Roche Molecular System (Brancburg, NJ). RESULTS: Viral load in untreated subjects showed wide variation for HBV, HCV and HIV. HBV has been demonstrated much stable to the conservation +4 degrees C also until 168 h while for HCV and HIV a greater decrease of the viral-load was observed. For all and three virus the conservation to +4 degrees C until 72 h does not seem to involve meaningful fall in the viral-load. A remarkable reduction of the viral-load has been observed after five cycles of freezing and defrosting. All the tests showed a good robustness to cross-contamination. The detection-limit (95%) was 8 U/ml for HBV, 21 U/ml for HCV and 27 copy/ml for HIV. CONCLUSIONS: Samples for NAT testing, can be stored until 72 h to +4 degrees C without appreciable lowering of the viral-load. Repeated cycles of changes of state should be avoided. The tests showed a good robustness to cross-contamination. NAT tests for biological qualification of blood units had a minimal sensibility around 50 (copy/unit/ml). In our experience the detection-limit (95%) was 21 U/ml for HCV, 27 copies/ml for HIV, 8 U/ml for HBV. The availability of NAT test for HBV-DNA, HCV-RNA e HIV-RNA, sensitive and reliable, together with epidemiological data, suggest the opportunity to place side by side, in the biological qualification of the blood units, to add the tests for HBV-DNA and HIV-RNA to the test for HCV-RNA mandatory by low, in Italy in the biological qualification of blood units.

[Foetal-maternal alloimmunizations in the South-East area of the Venice province]. / Alloimmunizzazioni feto-materne nel Sud-Est del Veneto.

BACKGROUND: This study report the results obtained in a retrospective analysis of the foetal-maternal alloimmunizations observed from 1993 to 1999 in the South-East area of the Venice province. METHODS: The data concerning 17,000 pregnancy observed in this area from 1993-1999 have been collected. For each pregnancy data concerning maternal ABO, Rh, Kk and IAT as well as foetal ABO, Rh, Kk and DAT were available. Further data (mainly antibodies concentration and specificity) were available if a foetal-maternal alloimmunization was detected and if transfusional support was given after the birth. RESULTS: The authors observed 465 alloimmunizations (prevalence 2.7%): 381 (82%) of these were due to an ABO foetal-maternal incompatibility, 23 due to D incompatibility and the other 61 due to other blood groups antigens. Only 6 cases needed transfusional support: 5 exchange transfusion (a patient needed 2 exchanges) and a delayed transfusion. CONCLUSIONS: Foetal-maternal alloimmunizations are today a rare but not exceptional event (about 3% of pregnancy), the great majority of these alloimmunizations are due to an ABO incompatibility. Despite the prevention of alloimmunization in D negative women by using Rh immune globulin, anti-D alloimmunization is still observed. A great number of other blood groups antigens are involved in foetal-maternal alloimmunization mainly within the Rh system (CcEe, etc.). In the authors' experience the great majority of foetal-maternal alloimmunizations were clinically silent, only 6 cases (1.3% of patients with a positive DAT) needed transfusional therapy.

The stability of hepatitis C virus RNA after storage at +4 degrees C.

One of the major issues in nucleic acid testing is how to store blood samples to obtain reliable results. We therefore studied hepatitis C virus (HCV) RNA concentration in samples after storage at +4 degrees C or freezing and thawing. Six HCV RNA-positive, untreated subjects were studied. Blood samples were collected from these subjects in plasma preparation tubes. The HCV RNA concentration was analysed after storage at +4 degrees C for 168 h or after five freeze-thaw cycles. For HCV RNA quantification we used a qualitative and a quantitative commercial test. After 168 h of storage at +4 degrees C, the HCV RNA concentration was similar to that observed at time-point 0 (5.025 log vs 5.492 log). In one sample we observed a significant fall in HCV RNA concentration. After five freeze-thaw cycles, the HCV RNA concentration was lower than that observed at time-point 0 (4.454 log vs 5.492 log) and in four samples we observed a significant fall in HCV RNA concentration. Our data suggest that HCV RNA is stable in whole blood samples stored at +4 degrees C for 168 h. Based on our results, we conclude that the standard procedures for transport of blood samples (at room temperature for a maximum of 5 h) and storage schedules (+4 degrees C for a maximum of 48 h) can be maintained without compromising the quality of results.