Transplanted human pancreatic islets after long-term insulin independence.

Long-term insulin independence after islets of Langerhans transplantation is rarely achieved. The aims of this study were to identify the histological and immunological features of islets transplanted in a type 1 diabetic patient who died of a cerebral hemorrhage after >13 years insulin independence. Islets were pooled from two donors with respectively one and five HLA mismatches. Insulin-positive islets were found throughout the right and left liver, and absent in the pancreas. Two- and three-dimensional analysis showed that islets lost their initial rounded and compact morphology, had a mean diameter of 136 µm and were constituted of an unfolded epithelial band of 39.1 µm. Leukocyte phenotyping showed no evidence of a tolerogenic environment in the islet-containing portal spaces. Finally, HLA typing of microdissected islets showed HLA from the best matched donor in all 23 microdissection samples, compared to 1/23 for the least matched donor. This case report demonstrates that allogeneic islets can survive over 13 years while maintaining insulin independence. Allogeneic islets had unique morphologic features and implanted in the liver regardless of their size. Finally, our results suggest that, in this case, rejection had been prevalent over autoimmunity, although this hypothesis warrants further investigation.

A novel stroke locus identified in a northern Sweden pedigree: linkage to chromosome 9q31-33.

OBJECTIVES: The population of northern Sweden is characterized by reduced genetic diversity and a high incidence of stroke. We sought to reduce genetic variation further, using genealogic analysis in a set of nuclear families affected by stroke, and we subsequently performed a genome-wide scan to identify novel stroke susceptibility loci. METHODS: Through genealogy, 7 nuclear families with a common ancestor, connected over 8 generations, were identified. A genome-wide scan using 449 microsatellite markers was performed with subsequent haplotype analyses. RESULTS: A maximum allele-sharing lod score of 4.81 on chromosome 9q31-q33 was detected. Haplotype analysis identified a common 2.2-megabase interval in the chromosomal region in 4 of the nuclear families, where an overrepresentation of intracerebral hemorrhage was observed. CONCLUSIONS: We have identified a novel susceptibility locus for stroke. Haplotype analysis suggests that a shared genetic factor is of particular importance for intracerebral hemorrhage.

Imaging the pancreas: from ex vivo to non-invasive technology.

While many recently published reviews have covered non-invasive nuclear imaging techniques, the aim of this review is to focus on current developments in optical imaging technologies for investigating the pancreas. Several of these modalities are being developed into non-invasive, real-time monitoring routines for pancreatic diseases. However, they also provide pre-clinical ex vivo and/or intravital tools for three-dimensional quantitative assessments of cellular and molecular events, with levels of specificity and resolution difficult to achieve with other currently available modalities.

The biomechanical properties of the feline femur.

The purpose of this study was to determine the biomechanical properties of feline long bone by testing cadaver bone from mature cats in compression, three-point bending, notch sensitivity and screw pull-out strength. The determination of these properties is of clinical relevance with regard to the forces resulting in long bone fractures in cats as well as the behaviour and failure mode of surgical implants utilized for fracture stabilization and repair in the cat. Cadaveric cat femurs were tested in compression, three-point bending and in three-point bending after the addition of a 2.0 mm screw hole. Cortical screws, 2.7 mm in diameter, were inserted in cadaveric cat femur samples for screw pull-out testing. The mean maximum load to failure of mid diaphyseal feline femurs tested in compression was 4201+/-1218 N. Statistical analysis of the parameter of maximum load tested in compression revealed a statistical difference between sides (p=0.02), but not location (p=0.07), or location by side (p=0.12). The maximum strength of mid diaphyseal feline femurs tested in compression was 110.6+/-26.6 MPa. The modulus of elasticity of mid-diaphyseal cat femurs tested in compression was determined to be 5.004+/-0.970 GPa. The mean maximum load to failure of feline femurs tested in three-point bending was 443+/-98 N. The mean maximum load to failure of feline femurs tested in three-point bending after a 2.0 mm diameter hole was drilled in the mid-diaphyseal region of each sample through both cortices was 471+/-52 N. The mean maximum load required for screw pull-out of 2.7 mm cortical screws placed in feline femurs tested in tension was 886+/-221 N. This data should be suitable for investigating fracture biomechanics and the testing of orthopaedic constructs commonly used for fracture stabilization in the feline patient.

Cryohypophysectomy used in the treatment of a case of feline acromegaly.

A 10-year-old female spayed cat was diagnosed with acromegaly secondary to a pituitary tumour. At the time of diagnosis, the cat had insulin-resistant diabetes mellitus and its insulin-like growth factor-I levels were elevated. Clinical signs included polyuria, polydipsia and weight gain. Persistent hyperglycaemia and glucosuria were identified, and fructosamine levels remained elevated. Magnetic resonance imaging of the brain showed a pituitary tumour. Transsphenoidal cryohypophysectomy was used to treat the pituitary tumour. Postoperatively, the serum insulin-like growth factor-I levels decreased and the diabetes mellitus was controlled with routine levels of insulin. To the authors' knowledge, this is the second reported case of acromegaly treated with cryohypophysectomy, and the first that reports a favourable long-term outcome. Cryohypophysectomy may be a safe and effective treatment for cats with a pituitary mass resulting in acromegaly.

Evaluation of a short glass fibre-reinforced tube as a model for cat femur for biomechanical testing of orthopaedic implants.

The biomechanical testing of tubes made of third generation short glass fibre-reinforced (SGFR) material approximating cat femurs was performed in order to determine their suitability as cat femur surrogates for the biomechanical testing of orthopaedic implants. The tubes were tested in compression, three-point bending, notch testing, and screw pullout. Thin walled (B1-tubes) had a 13% lower maximum load to failure, a 19% higher maximum strength and a 13% lower elastic modulus compared to cat femurs tested in compression. B1-tubes maximum load to failure in three-point bending and screw pullout strength were considerably lower compared to cat femurs (29% and 63%, respectively). Notch testing was not performed on B1-tubes due to low bending strength. Thicker walled (B2-tubes) had a 23% higher maximum load to failure, a 10% higher maximum strength and a 21% lower elastic modulus compared to cat femurs tested in compression. The comparison of B2-tubes and cat femurs in three-point bending revealed a 7% increase in maximum load to failure for the B2-tubes. Drilled B2-tubes (notch testing) were weaker with a 30% lower load to failure compared to cat femurs. A screw pullout comparison of B2-tubes and cat femurs revealed a 2% increase in maximum load to failure for the B2-tubes. These tubes were intended to provide a model as a suitable surrogate for cat femurs for testing the bending strength of various orthopaedic constructs involving plates and screws. Testing revealed that third generation SGFR tubes were not suitable for these purposes and emphasizes the need to carefully evaluate the suitability of any model.

Lack of aggregation of ischemic stroke subtypes within affected sibling pairs.

OBJECTIVE: To establish whether subtypes of ischemic stroke aggregate within ischemic stroke-affected sibling pairs more than expected by chance alone. METHODS: This retrospective family study was based on a pooled analysis of two cohorts of male and female adult sibling pairs with symptomatic ischemic stroke. One hospital-based cohort of 404 individuals (first proband seen August 30, 1999) was recruited from the United States and Canada, and another population-based cohort of 198 individuals (first proband seen April 17, 1997) was recruited from Umeå, Sweden. Subtype diagnoses were based on Trial of Org 10172 in Acute Stroke Treatment (TOAST) criteria. RESULTS: Agreement for subtype diagnoses within families was poor (mean +/- asymptotic SE kappa = 0.17 +/- 0.04). Occurrence of one ischemic stroke subtype in a proband was not associated with a greater likelihood of that subtype being the qualifying stroke subtype in the sibling. Comparable levels of agreement were seen when restricting the analysis to same-sex sibling pairs (kappa = 0.22 +/- 0.05) to sibling pairs in which the proband's stroke occurred before the age of 65 years (kappa = 0.16 +/- 0.05) or to pairs in which the proband's stroke occurred at or after the age of 65 years (kappa = 0.19 +/- 0.05). CONCLUSIONS: The subtype of ischemic stroke in a proband was a poor determinant of the subtype of ischemic stroke in the respective sibling. This suggests that many genetic risk factors for ischemic stroke may not be specific for one subtype.

Genetic control of parasite clearance leads to resistance to Plasmodium berghei ANKA infection and confers immunity.

Unprecedented cure after infection with the lethal Plasmodium berghei ANKA was observed in an F2 progeny generated by intercrossing the wild-derived WLA and the laboratory C57BL/6 mouse strains. Resistant mice were able to clear parasitaemia and establish immunity. The observed resistance was disclosed as a combinatorial effect of genetic factors derived from the two parental strains. Genetic mapping of survival time showed that the WLA allele at a locus on chromosome 1 (colocalizing with Berghei resistance 1 (Berr1), a locus associated with resistance to experimental cerebral malaria) increases the probability to resist early death. Also, the C57Bl/6 allele at a novel locus on chromosome 9 (Berr3) confers overall resistance to this lethal Plasmodium infection. This report underlines the value of using wild-derived mouse strains to identify novel genetic factors in the aetiology of disease phenotypes, and provides a unique model for studying parasite clearance and immunity associated with malaria.

Identification of a susceptibility locus for migraine with and without aura on 6p12.2-p21.1.

Migraine is the most common type of chronic episodic headache. To find novel susceptibility genes for familial migraine with and without aura, a genomewide screen was performed in a large family from northern Sweden. Evidence of linkage was obtained on chromosome 6p12.2-p21.1, with a maximum two-point lod score of 5.41 for marker D6S452. The patients with migraine shared a common haplotype of 10 Mb between markers D6S1650 and D6S1960.

Low rate of proliferation in immature thymocytes of the non-obese diabetic mouse maps to the Idd6 diabetes susceptibility region.

AIMS/HYPOTHESIS: The non-obese diabetic (NOD) mouse spontaneously develops T-cell-dependent autoimmune diabetes. This mouse strain has a number of immune dysfunctions related to T-cell development but so far there are no available data on the proliferation of NOD immature thymocytes. We therefore studied the thymocyte proliferation in the NOD mouse in discrete stages of T-cell development. METHODS: We depleted thymocytes in vivo and analysed thymocyte proliferation during the thymus recovery from depletion. We used co-segregation analysis and quantitative loci trait analysis to investigate the genetic control of proliferation impairments in NOD thymocytes. RESULTS: Immature thymocytes of female NOD mice proliferate with a relatively low rate compared to non-autoimmune C57Bl/6 mice. This aberrant proliferation was most pronounced in CD4-/lo CD8+ cells differentiating from the CD4-CD8- to the CD4+CD8+ stage. A genetic mapping study using an F2 intercross between the NOD and the C57BL/6 strains showed that a major locus controlling this trait is linked to the insulin-dependent diabetes susceptibility locus Idd6. CONCLUSION/INTERPRETATION: Our results suggest that impairment of proliferation of immature thymocytes is one possible mechanism through which the Idd6 locus contributes to the pathogenesis of diabetes.

Nasal dermoid sinus in an American cocker spaniel.

An American cocker spaniel was presented for a subcutaneous mass and draining tract located between its eyes. Contrast radiography and surgical excision showed communication of the tract with the left frontal sinus and rostral cerebral dura, respectively. A dermoid sinus was diagnosed by a combination of gross and histologic findings.

CTLA-4-/- mice display T cell-apoptosis resistance resembling that ascribed to autoimmune-prone non-obese diabetic (NOD) mice.

The genes conferring susceptibility to autoimmune (insulin-dependent) diabetes mellitus (IDDM) are, in most cases, not defined. Among the loci so far identified as associated with murine IDDM (Idd1-19), only the nature of Idd1 has been assessed. Here we show that thymocytes and peripheral lymphocytes of the non-obese diabetic (NOD) mouse are relatively resistant to apoptosis induced by gamma-irradiation. By linkage analysis of F2 progeny mice, we map this trait to a locus on chromosome 1 containing the Idd5 diabetes susceptibility region. By the use of congenic mice, we confirm the linkage data and map this locus to a 6 cM region on proximal chromosome 1. Ctla4, being localized in this chromosomal region and mediating crucial functions in T cell biology, is a logical candidate gene in the Idd5 susceptibility region. In line with this, we demonstrate that T cells from Ctla4(-/-)deficient mice show a similar resistance to gamma-irradiation-induced apoptosis as observed in the NOD mice. This reinforces the notion that CTLA-4 contributes to the pathogenesis of autoimmune diabetes.

Use of pericardial patch graft reconstruction of the right atrium for treatment of hemangiosarcoma in a dog.

Atrial mass resection is possible in a limited number of dogs with hemangiosarcoma in which only the right atrial appendage or atrial free wall is involved, A pericardial patch graft can be used to reconstruct the right atrium after resection of large atrial tumors. Tumor-free margins can be obtained by use of this technique.

The basic helix-loop-helix transcription factor E2-2 is involved in T lymphocyte development.

E2A, HEB and E2-2 genes encode a group of basic helix-loop-helix (bHLH) transcription factors that are structurally and functionally similar. Deletion of the genes encoding either of these proteins leads to early lethality and a block in B lymphocyte development. Evidence for a function in T lymphocyte development has, however, only been reported for E2A and HEB. To further elucidate the role of E2-2 at developmental stages that have proven difficult to study due to the early lethality phenotype of mice defective in E2-2, we generated and analyzed mice conditionally mutated in the E2-2 gene. These mice are mosaic with respect to E2-2 expression, consisting of cells with either one functional and one null mutated E2-2 allele or two null mutated alleles. Using this experimental model, we find that cells with a homozygous null mutated E2-2 gene are under-represented in B lymphocyte as well as T lymphocyte cell lineages as compared to other hematopoietic or non-hematopoietic cell lineages. Our data suggests that E2-2 deficiency leads to a partial block in both B and T lymphocyte development. The block in T cell development appears to occur at an early stage in differentiation, since skewing in the mosaicism is observed already in CD4+8+ double-positive thymocytes.

Functional expression of Cre recombinase in sub-regions of mouse CNS and retina.

We describe a strategy for generating CNS and retina sub-region-specific mutations using the Cre/loxP system. Transgenic mice expressing Cre recombinase under the control of the c-kit promoter were established. Functional Cre expression was predominantly found to be restricted to the CA1, CA2 and CA3 regions of the hippocampus, the anterior region of the dentate gyrus, and to the ganglion cell layer of the retina.

Kidney transplantation in dogs with naturally occurring end-stage renal disease.

Renal allografts were performed between unrelated donors and 15 dogs with naturally occurring end-stage renal disease. Donor selection was based on compatible dog erythrocyte antigen typing and cross-matching. An immunosuppressive protocol consisting of rabbit antidog antithymocyte serum, cyclosporin-A, azathioprine, and prednisone was used to control postoperative rejection of the donated kidney. Although seven animals died because of technical failures or rejection episodes, a median survival time of eight months has been achieved, with two dogs living for longer than five years after surgery. Long-term survivors have died from a variety of problems not related to renal allograft rejection.