The role of human demographic history in determining the distribution and frequency of transferase-deficient galactosaemia mutations.

Classical or transferase-deficient galactosaemia is an inherited metabolic disorder caused by mutation in the human Galactose-1-phosphate uridyl transferase (GALT) gene. Of some 170 causative mutations reported, fewer than 10% are observed in more than one geographic region or ethnic group. To better understand the population history of the common GALT mutations, we have established a haplotyping system for the GALT locus incorporating eight single nucleotide polymorphisms and three short tandem repeat markers. We analysed haplotypes associated with the three most frequent GALT gene mutations, Q188R, K285N and Duarte-2 (D2), and estimated their age. Haplotype diversity, in conjunction with measures of genetic diversity and of linkage disequilibrium, indicated that Q188R and K285N are European mutations. The Q188R mutation arose in central Europe within the last 20 000 years, with its observed east-west cline of increasing relative allele frequency possibly being due to population expansion during the re-colonization of Europe by Homo sapiens in the Mesolithic age. K285N was found to be a younger mutation that originated in Eastern Europe and is probably more geographically restricted as it arose after all major European population expansions. The D2 variant was found to be an ancient mutation that originated before the expansion of Homo sapiens out of Africa.

Protective role of the L-arginine-nitric oxide synthase pathway on preservation injury after rat liver transplantation.

BACKGROUND: A major problem complicating liver transplantation is the preservation injury that results from cold storage and subsequent ischemia/reperfusion injury after organ revascularization. The L-arginine-nitric oxide (NO) pathway has been recognized to play critical roles during infection, inflammation, organ injury, and transplant rejection. Recent data indicates that NO synthesis has beneficial effects in several models of liver injury. The purpose of this study is to examine the role of the L-arginine-NO pathway on preservation injury in an experimental model of rat liver transplantation. METHODS: Orthotopic liver transplantation was performed in syngeneic (LEW to LEW) rats. Liver preservation injury was determined by measuring serum liver function tests 6 to 48 hours after transplantation. In some experiments, rats received L-arginine supplementation 0 to 24 hours after transplantation. In other experiments, NO synthase inhibitors (L-NAME or L-NIL) were injected at the time of isograft revascularization. RESULTS: L-Arginine supplementation decreased hepatic transaminase levels at all time points examined (6-48 hours). L-Arginine produced a significant improvement in liver preservation injury by 12 hours after reperfusion. The NO synthase inhibitor L-NAME caused a significant increase in liver injury 24 hours after injection. The inducible NO synthase (iNOS)-specific inhibitor L-NIL had no significant effect on liver injury. CONCLUSIONS: The results show that L-arginine supplementation and NO synthesis improve hepatic injury and have a protective role in the transplanted liver graft. The protective effect may be mediated by low-level cNOS-derived NO.

Spontaneous pelvic hematoma simulating neoplasm: case report and literature review.

Spontaneous hematomas are rare and can present with acute or chronic symptoms. Our patient presented with deep vein thrombosis of the lower extremity associated with a spontaneous pelvic hematoma. This lesion was radiologically and clinically indistinguishable from a soft-tissue neoplasm. The case of a spontaneous pelvic neoplasm in an otherwise healthy young man is presented and the literature reviewed regarding issues of differential diagnosis and clinical management.

Adenovirus-mediated gene transfer to liver grafts: an improved method to maximize infectivity.

BACKGROUND: Adenoviral gene therapy in liver transplantation has many potential applications, but current vector delivery methods to grafts lack efficiency and require high titers. In this study, we attempted to improve gene delivery efficacy using three different delivery methods to liver grafts with adenoviral vector encoding the LacZ marker gene (AdLacZ). METHODS: AdLacZ was delivered to cold preserved rat liver grafts by: (1) continuous perfusion via the portal vein (portal perfusion), (2) continuous perfusion via both the portal vein and hepatic artery (dual perfusion), and (3) trapping viral perfusate in the liver vasculature by clamping outflow (clamp technique). RESULTS: Using 1x10(9) plaque-forming units of Ad-LacZ (multiplicity of infection of 0.4), transduction rate in 3-hr preserved liver grafts, determined by 5-bromo-4-chromo-3-indolyl-beta-D-galactopyranoside staining and beta-galactosidase assay 48 hr after transplantation, was best with clamp technique (21.5+/-2.7% 5-bromo-4-chromo-3-indolyl-beta-D-galactopyranoside-positive cells and 81.1+/-3.6 U/g beta-galactosidase), followed by dual perfusion (18.5+/-1.8%, 66.6+/-19.4 U/g) and portal perfusion (8.8+/-2.5%, 19.7+/-15.4 U/g). Further studies using clamp technique demonstrated a near-maximal gene transfer rate of 30% at multiplicity of infection of 0.4 with prolonged cold ischemia to 18 hr. Transgene expression was stable for 2 weeks and slowly declined to 7.8+/-12.1% at day 28. Lack of inflammatory response was confirmed by histopathological examination and liver enzymes. Transduction was selectively induced in hepatocytes with nearly no extrahepatic transgene expression in the lung and spleen. CONCLUSIONS: The clamp technique provides a highly efficient viral gene delivery method to cold preserved liver grafts. This method offers maximal infectivity of adenoviral vector with minimal technical manipulation.

The child, his family and dyspraxia.

Children with dyspraxia have difficulties with co-ordination, eg poor balance or problems doing up buttons or writing with a pencil. The cause is unknown. Though mentally normal and with no known neurological condition, these children are unable to plan, organise and co-ordinate their movements. School work is affected and occupational therapy and physiotherapy may be needed. A supportive atmosphere helps counter poor self-esteem. As this study shows, the condition puts strains on the parents and the whole family. Support from the health visitor and school nurse would often be appreciated. With practice, children with dyspraxia can achieve reasonable degrees of motor skills. Children do not necessarily grow out of the condition but can be helped with empathy and appropriate teaching and therapy.

Evaluating groups in learning disabilities.

Groupwork can be effective in meeting a range of needs presented by students with profound learning disabilities. This article describes the process involved in setting up groups for these students, and includes examples of a group session and methods for evaluating groupwork.

Murine cytomegalovirus-associated arteritis.

Unique inflammatory lesions affecting the ascending aorta and pulmonary artery of BALB/c and C57BL/6 mice infected with murine cytomegalovirus (MCMV) were identified in a pilot and two subsequent experiments to characterize the potential effect of MCMV infection on diet-induced atherosclerotic lesions. Suckling BALB/c and C57BL/6 mice were inoculated with MCMV and subsequently fed either a commercial mouse diet or a synthetic atherogenic diet from weaning. The three experiments varied with respect to the age of the mice at the time of MCMV inoculation and the dose of virus given. The conditions of MCMV exposure were progressively modified in the three experiments to increase the prevalence of MCMV-associated inflammatory lesions in the pulmonary artery and aorta. In the final experiment, in which suckling mice were inoculated at 9 days of age, MCMV-associated arteritic lesions had an observed prevalence at 8 weeks post-inoculation of 87.5% (7/8) in BALB/c mice on the normal diet and 100% (8/8) in C57BL/6 mice on the normal diet and in both strains on the atherogenic diet. The inflammatory lesions in both vessels were characterized by mononuclear cell infiltrates containing CD3+, CD4+, and CD8+ lymphocytes. The cellular infiltrates were often more intense on the adventitial surface and infiltrated into the overlying tunica media. The intima was infiltrated by mononuclear cell infiltrates that appeared to contain more macrophages and fewer lymphocytes than did the adventitial infiltrates.(ABSTRACT TRUNCATED AT 250 WORDS)