Kidney transplant monitoring by anti donor specific antibodies

Donor-specific anti-HLA antibodies were studied by cytotoxicity crossmatching (CTXM) and flow cytometry crossmatching (FCXM) in 117 kidney transplant candidates; the same study was carried out in 33 cadaver-donor kidney recipients, during the first 3 post-transplant months, for which donor cells were available. Pre-transport evaluation showed that 82.9 % of subjects were CTXM negative/FCXM negative, 6.8 % of patients were positive in both tests, and 10.3 % were CTXM negative/FCCM positive. Post-transplant monitoring for donor-specific antibodies (Abs-DS) showed that nine recipients (27.3 %) were FCXM positive; six of them were IgG + and three IgM +. In comparing these results with the clinical course, a significant association between FCXM IgG + and rejection episodes was observed (P < 0.01).     

How four and twelve weeks of implantation affect the strength and stiffness of a tendon graft securely fixed in a bone tunnel: a study of Evolgate fixation in an extra-articular model ovine model

Healing of a tendon graft to a bone tunnel is slower than the healing of a bone plug. Therefore, the device chosen for hamstring fixation may need to maintain its strength and stiffness longer than the device chosen for bone-tendon-bone fixation. We evaluated, in an extraarticular ovine model, how 4 and 12 weeks of implantation affect the strength of a tendon graft fixed to bone with the Evolgate. The long digital extensor tendon was transplanted and fixed with the Evolgate into a 30-mm long, 8 mm diameter bone tunnel drilled in the tibial metaphysis of both posterior limbs of 15 skeletally mature Suffolk sheep. Immediately after implantation, and 4 and 12 weeks later, biomechanical cyclic load tests in 50 N increments were performed until failure to evaluate the ultimate failure load (UFL). Histological analysis was also performed at 4 and 12 weeks. Biomechanical tests revealed a UFL of 339±120 N at time 0, and increases to 635±19 N (4 weeks) and to 867±80 N (12 weeks). The differences between all 3 groups were significant (p<0.001, paired t test). The histological evaluation showed a layer of cellular, fibrous tissue between the tendon and the bone, along the length of the bone tunnel; this layer progressively matured and reorganized during the healing process. The collagen fibers that attached the tendon to the bone resembled Sharpey’s fibers. The strength of the interface significantly and progressively increased between weeks 4 and 12 after transplantation, and was associated with a degree of bone ingrowth noted histologically. The use of the Evolgate seems not to interfere with the bone ingrowth after implantation, allowing an improvement in strength of the bonetendon- device complex.     

Cell mediated immunity (CMI) and post transplant viral infections — Role of a functional immune assay to titrate immunosuppression

Cell mediated immunity (CMI) was assessed by the ImmuKnow assay in 12 patients after kidney transplantation, who presented with viral infection. Treatment included lowering of immunosuppression in all cases and antiviral treatment if indicated. The assay was repeated during the follow up. The ImmuKnow assay at time of presentation of viral infections was 56.8 ± 58.2 (range 3–178; median 22) ATP ng/ml. With the clearance of viral infection and lowering of immunosuppression, the assay showed an increase in the level of CMI at 194.5 ± 118.9 (range 53–409; median 150) ATP ng/ml. There was viral clearance or stabilization in all cases and there was no incidence of allograft rejection. The ImmuKnow assay of CMI can be used to titrate initial immunosuppression reduction and its subsequent increase, in patients with viral infection after transplantation.     

Telomerase activity in prognostic histopathologic features of melanoma.

BACKGROUND: Telomerase activity (TA) is believed to play a role in the regulation of senescence and to limit the number of cell divisions. The deregulation of telomerase appears to contribute to oncogenesis and the formation of immortal cell lines. As a result, it is believed that it could be used as a prognostic marker in melanoma. METHODS: TA was assayed by the polymerase chain reaction PCR-ELISA-based telomeric repeat amplification protocol (TRAP assay). One hundred and eight samples were distributed in four histological groups: 30 samples from primary cutaneous melanomas, 24 from peritumoural skin sites, 28 from benign melanocytic lesions, and 26 from normal skin sites as a control. RESULTS: TA was different among the four tested groups (Kruskall-Wallis test p<0.001), and increasing values of TA were observed progressing from normal skin to benign and then to malignant lesions. Among melanoma samples, there was a significant association between TA and ulceration (p=0.025), TA and vascular invasion (p=0.018) and TA and mitotic rate (p=0.029) (Mann-Whitney test). A linear regression analysis showed significant associations between the increase of TA with Breslow thickness (p=0.004) and the presence of satellites (p=0.002). CONCLUSIONS: We observed that TA had increased from control skin to peritumoural skin, and then to benign melanocytic lesions and finally to melanoma, suggesting tumour progression. TA showed higher values in the presence of some important histopathologic parameters related to poor prognosis in cutaneous melanoma such as ulceration, vascular invasion, satellites, high rates of mitosis, and in thicker tumours.     

Inhibition by N-acetylcysteine of carcinogen-DNA adducts in the tracheal epithelium of rats exposed to cigarette smoke

The ability of the aminothiol N-acetylcysteine (NAC) to preventthe formation of carcinogen-DNA adducts in tracheal epithelialcells was investigated in Sprague-Dawley rats exposed whole-bodyto mainstream cigarette smoke for either 40 or 100 consecutivedays. ³²P-Postlabelling analyses showed the occurrence of DNAadducts (12.49 adducts/10⁸ nucleotides) after 40 days of exposure,with a trend to formation of characteristic diagonal radioactivezones. Total adduct levels were not further enhanced after 100days of exposure to smoke, although significant changes occurredin the amounts of individual adducts. NAC, given by gavage inthe 40 day study and in drinking water in the 100 day study,significantly inhibited the formation of smoke-related carcinogen-DNAadducts in the tracheal epithelium, to such an extent that adductlevels were not significantly higher than those detected insham-exposed control rats. Together with a variety of othermolecular, clastogenicity, metabolic, cytological and histopathologicalend-points investigated in rodents and with the preliminaryevidence arising from a study in humans, these results documentthe considerable efficacy of oral NAC in inhibiting smoke-relatedcarcinogen-DNA adducts.     

Characterization of a YAC containing part or all of the Norrie disease locus

It has been shown from pulsed-field gel electrophoresis (PFGE)that the monoamine oxidase genes A and B (MAOA & MAOB) andDXS7 loci are physically very close. We have therefore extendedstudies on their relationship through the characterisation ofa 650 kb YAC isolated using L1.28 (recognising the DXS7 locus)as a probe. Restriction mapping of the YAC indicates that itcontains both MAOA and MAOB genes in addition to the DXS7 locus.The map derived from the YL1.28-YAC is compatible both withthe map from an independently derived YAC carrying MAOA andB genes and with the long range genomic map for the region.A series of subclones prepared from a 'phage library (lambdaDASH II) of the YAC have been characterised and have been employedto determine the end point of the deletion of a Norrie disease(NDP) patient who has been shown to lack both DXS7 and MAO codingsequences. The pattern of retention of subclones in the deletionpatient place the end point of the deletion within 30–130kb of the proximal end of the YAC. By combining the data withestablished recombination analysis, we provide evidence thatall or part of the NDP lies in the interval of approximately250kb within the YAC.     

CXCL10 haplotypes and multiple sclerosis: association and correlation with clinical course

CXCL10 (interferon- γ -inducible protein-10) levels are increased in cerebrospinal fluid of multiple sclerosis (MS) patients with symptomatic attacks of inflammatory demyelination, supporting a role for this molecule in MS pathogenesis. Two hundred and twenty-six patients with MS and 235 controls were genotyped for G  →  C and T  →  C single nucleotide polymorphisms (SNPs) in exon 4 of CXCL10 gene. Haplotypes were tested for association and correlated with clinical variables. The two SNPs studied were in complete linkage disequilibrium. None of the determined haplotypes was associated with MS. However, carriers of the GGTT haplotype (defined as wild type, according to the sequence in National Centre for Biotechnology Information (NCBI) database) had a significantly lower progression index than non-carriers ( P  = 0.016). Furthermore, amongst patients who had an initial relapsing remitting (RR) course of the disease, the time between onset and second episode was significantly longer in GGTT carriers ( P  = 0.021). Considering secondary progressive (SP)–MS patients, the time between the initial RR form and the subsequent worsening to SP was longer in this group ( P  = 0.08). Therefore, the GGTT haplotype of the CXCL10 gene is not a susceptibility factor for the development of MS, but is probably to influence the course of MS, possibly contributing to slow down the progression of the disease.     

Preemptive Plasmapheresis and Recurrence of FSGS in High-Risk Renal Transplant Recipients

Recurrent focal segmental glomerulosclerosis (FSGS) following transplantation is ascribed to the presence of a circulating FSGS permeability factor (FSPF). Plasmapheresis (PP) can induce remission of proteinuria in recurrent FSGS. This study addressed the efficacy of pre-transplant PP in decreasing the incidence of recurrence in high-risk patients. Ten patients at high-risk for FSGS recurrence because of rapid progression to renal failure (n = 4) or prior transplant recurrence of FSGS (n = 6) underwent a course of 8 PP treatments in the peri-operative period. Recurrences were identified by proteinuria >3 g/day and confirmed by biopsy. Seven patients, including all 4 with first grafts and 3 of 6 with prior recurrence, were free of recurrence at follow-up (238-1258 days). Final serum creatinine in 8 patients with functioning kidneys averaged 1.53 mg/dL. FSGS recurred within 3 months in 3 patients, each of whom had lost prior transplants to recurrent FSGS. Two of these progressed to end-stage renal disease (ESRD) and the third has significant renal dysfunction. Based on inclusion criteria, recurrence rates of 60% were expected if no treatment was given. Therefore, PP may decrease the incidence of recurrent FSGS in high-risk patients. Definitive conclusions regarding optimal management can only be drawn from larger, randomized, controlled studies.