Comprehensive clinical, biochemical and genetic screening reveals four distinct GBA genotypes as underlying variable manifestation of Gaucher disease in a single family.

Gaucher disease (GD) is a lysosomal storage disorder that is associated with bi-allelic pathogenic variants in GBA. Its wide clinical spectrum, ranging from mild organomegaly to significant skeletal and neurological involvement, is partially explained by genotype-phenotype correlations. We present a family, in which all members over two generations presented with at least splenomegaly. Comprehensive clinical, biochemical and genetic workup was required to diagnose GD, which is caused by as many as four distinct GBA genotypes.

The first case of primary alpha-sarcoglycanopathy identified in Albania, in two siblings with homozygous alpha-sarcoglycan mutation.

Limb-girdle muscular dystrophy type 2D (LGMD2D) is caused by autosomal recessive mutations in the alpha-sarcoglycan gene. The clinical, biochemical, histological, imunohistochemical and molecular genetic data in 2 Albanian siblings with LGMD2D (adhalinopathy or alpha-sarcoglycanopathy) are presented and the resemblance with Duchenne muscular dystrophy (DMD) is discussed. Both siblings had very high level of CK and a negative molecular test for DMD deletions and duplications. The muscle biopsy showed dystrophic features as well as deficiency in two different proteins, the Gamma sarcoglycan protein (-SG) and the Alpha -SG protein (-SG). DNA analysis demonstrated homozygosity for a pathogenic point mutation (574C>T) in the alpha-sarcoglycan gene, confirming the diagnosis of limb-girdle muscular dystrophy type 2D. We believe it is the first confirmed case of primary alpha-sarcoglycanopathy identified in Albania which support the assumption of a wide geographic prevalence of severe childhood onset of autosomal recessive muscular dystrophy, We show that muscle biopsy and DNA diagnosis remains the most sensitive and specific method for differential diagnosis.

Decontamination of heart valve and arterial allografts in the European Homograft Bank (EHB): comparison of two different antibiotic cocktails in low temperature conditions.

The aim of the study was to compare the efficiency of two different antibiotic cocktails in the cardiovascular allograft decontamination. Low temperature, low-concentration antibiotic cocktail with Cefoxitin, Lincomycin, Polymixin B and Vancomycin was decontamination protocol in EHB for many years. The modified cocktail doesn't contain Cefoxitin. The study had two steps. First step: cardiovascular allografts from 80 donors are incubated in classical (group 1) or modified cocktail (group 2). Second step: 184 and 182 allografts of group 1 and group 2 are incubated in the modified and classical antibiotic cocktail, respectively. The bacteriological examination is performed in three steps: A-transport solution, B-decontamination solution and C-cryopreservation solution. During the first step 23.75% of the tissues were initially contaminated mainly with Staphylococcus (78.95%). 93.75% of the allografts of group 1 and 100% of group 2 were sterile after incubation (p = 0.058). 25.54% and 30.77% of group 1 and 2, respectively were contaminated in A-examination during the second step. Staphylococci were isolated in 82.98% and 69.64% in group 1 and 2, respectively. About 4.35% of group 1 and 5.5% of group 2 were contaminated in A, B, and C whereas 5.4% of group 1 and 4.4% of group 2 were contaminated in B or C after being sterile in A. Finally 9.78% of the tissues were rejected and 90.22% cryopreserved in the modified, whereas 9.89% rejected and 90.11% accepted in the classical group (p = 0.1). The difference was non-significant in the level of decontamination between the two cocktails. Contamination of some tissues with low growing, low-pathogen germs that appeared in B or C examination, couldn't be explained. This issue needs complementary investigation.

Banking of the human heart valves and the arteries at the European homograft bank (EHB)--overview of a 14-year activity in this International Association in Brussels.

Processing of the human heart valves and arteries has been carried out at the European Homograft Bank (EHB) in Brussels since 1989 and 1991, respectively. Heart valve donors of 0-65 years were classified in (1) Beating heart donors (BHD), of which recipients of heart transplantation (RHT) and multiorgan donors (MOD) after brain death, and (2) non-beating heart donors (NBHD) with warm ischaemic time (WIT) of less then 6 h. Past history of the donors has been checked for malignant and chronic diseases, as well as biology for transmissible and infectious diseases. Perfect collaboration has been established with the transplant coordinators and transplant teams of the implanting centres. Dissection, decontamination, cryopreservation and storing in fluid nitrogen has been carried out in accordance with the Belgian and European Standards of cardiovascular allografts. During this period, a total of 2.828 hearts, 28 predissected valves and 616 batches of arteries arrived in the EHB. 3.537 valves and 1.137 different arteries were accepted for implantation. The main reasons for tissue rejection were morphology, contamination and cuts during the tissue retrieval or dissection. A huge network of different hospitals in Belgium and elsewhere in Europe and Switzerland were included in this process. Pulmonary allografts were not sent for implantation in the left ventricular outflow tract after 1998, since the early and mid-term results after 76 implantations were disappointing. The number of implanted aortic and pulmonary allografts remains stable from year to year, however the number of the allografts used for Ross operation is still increasing. Since the results of the follow up were disappointing, we still only require the implantation and immediate postoperative results, whereas the follow-up information only for specific study purposes.

Processing of cardiovascular allografts: effectiveness of European Homograft Bank (EHB) antimicrobial treatment (cool decontamination protocol with low concentration of antibiotics).

To assess the effectiveness of antimicrobial treatment by using cool decontamination protocol with low concentration of antibiotics during processing of cardiovascular allografts, 948 allografts processed during a 2-year period were analysed. Five hundred and fourty one donors aged <62 years were classified in: multiorgan donors (MOD) with non-transplantable hearts; recipients of cardiac transplantation (RHT); and non-beating heart cadavers with a warm ischemic time of less than 6 h (NBHD). During processing three samples for bacteriology testing were taken A (sampling before decontamination); B (sampling after decontamination); C (sampling on the final product). Samples A were positive in 348 cases (36.4%), respectively 36% for MOD, 21.6% for RHT and 78.1% for NBHD. All the allografts were immersed in a cocktail of four antibiotics at 4 degrees C. After exposure to antibiotics the rate of decontamination of those with A positive was 90.4, 92.5, 82.5% respectively for MOD, RHT, NBHD. At the end of processing, 57 allografts (6%) were positive in B and/or C, 15 allografts remained contaminated with the same bacteria as in A, 42 were contaminated during processing. The overall rate of sterility in the end of processing is 94% and for each group this is: 95.4% for MOD, 96.8% for RHT and 86.3% for NBHD. Analysis shows that there is no influence of time of exposure in AB in the rate of decontamination for MOD and RHT. The most predominant germ in contamination is Coagulase Negative Staphylococcus (CNS) (53.4% alone, 8.9% with other bacteria). 83.3% of MOD; 88.5% of RHT were contaminated with one germ, while 40.4% of NBHD were contaminated with more than one.

A novel 7.4 kb mitochondrial deletion in a patient with congenital progressive external ophthalmoplegia, muscle weakness and mental retardation.

We present a patient with external ophthalmoplegia, bilateral ptosis, progressive muscle weakness with "ragged-red fibres" and mental retardation. Mitochondrial DNA analysis by Southern blot revealed heteroplasmy in muscle for a 7.4 kb deletion. In white blood cells, the deletion was only detectable by PCR. There was no evidence for duplications, nor for multiple deletions in the proband or siblings. PCR analysis did not reveal the presence of a mitochondrial DNA defect in the parents and siblings. Thus, there is no experimental support for a maternally inherited mitochondrial DNA deletion. We consider this a sporadic case with a de novo deletion. Diabetes and complaints of fatigue, also seen in this family, are probably coincidental. Mental retardation has been reported occasionally in patients with mitochondrial deletions, but is not common.