The OKT3 Antibody Response Study: a multicentre study of human anti-mouse antibody (HAMA) production following OKT3 use in solid organ transplantation

Human anti-murine antibody titres following patient exposure to the monoclonal antibody Orthoclone OKT3 (muromonab-CD3) are determined by laboratories using diverse analytical methods which are not standardized and whose concordance is not established. A multicentre study group therefore compared testing for IgG anti-OKT3 antibody among seven laboratories. A set of 270 sera was obtained from 30 heart, 30 kidney and 30 liver transplant recipients with no previous exposure to OKT3 who were receiving OKT3 for induction immunosuppression. Sera were collected from each patient prior to and at 24 +/- 2 days and 31 +/- 2 days following initial OKT3 exposure. Identical aliquots of all 270 sera were tested for IgG anti-OKT3 antibody by each laboratory. In addition, the limit of detection of each laboratory's method was estimated by titration of an affinity-purified IgG anti-OKT3 reference material of known concentration. Anti-OKT3 antibody formation differed greatly among the three organ groups. Cardiac patients demonstrated the least sensitization and almost exclusively lower titres, while kidney recipients had more frequent and higher titre antibody formation. Liver recipients yielded the highest sensitization rate and the most frequent high titre sera. Importantly, the seven laboratories differed widely in the number of pretreatment sera reported as positive (ranging from 0% to 41% among laboratories), the number of post-OKT3 sera reported as positive (17-63%), the number of post-OKT3 samples with titre > or = 1000 (2-31%), and the number of patients sensitized 19-69%). Concordance among laboratories was highly variable, with interlaboratory agreement ranging from 38% to 83% on the sample titres assigned to 180 post-OKT3 sera. Many of the discordant results were consistent with differences in the limit of detection of the analytical methods, which ranged from 0.19 microgram/ml to > or = 15 micrograms/ml, a nearly 100-fold difference among laboratories. This study demonstrated the presence of both good concordance and significant discordance among laboratories in determining human anti-mouse antibody titres, and demonstrated that common titre categories (100, 1000, 10,000) were not equivalent among laboratories. The level of concordance among methods should be considered when comparing anti-OKT3 antibody results from different centres and their correlation with clinical events. Universal comparative testing, patterned after proficiency testing programmes, is needed to assess differences among laboratories and to bring uniformity and a sound interpretative basis to this field of testing.     

Leptin levels do not change acutely with food administration in normal or obese subjects, but are negatively correlated with pituitary-adrenal activity

Modern liver surgery is based upon deep knowledge of the surgical anatomy of the liver and improvements with ultrasounds imaging techniques have provided multidimensional interpretations of the liver anatomy intraoperatively. The technical advances with real-time scanning combined with the pioneering efforts of the Japanese liver surgeons have permitted the dynamic adaptation of the functional liver anatomy to the real anatomy, thus intimately aiding in segment oriented anatomical resection. Intraoperative ultrasound afford several advantages such as viewing the internal anatomy in direct relationship to the surface landmarks under the probe, enabling higher frequencies resulting in greater image resolution. The utilization of intraoperative ultrasound can modify the tactics involved with resection of both primary and metastatic liver tumours. The routine use of intraoperative ultrasound is strongly advocated since more complex procedures can be performed safely since the surgeon proceeds with complete knowledge of the real liver anatomy when deciding the feasibility and extent of liver resection.     

Stereological estimation of the total volume of MR visible brain lesions in patients with multiple sclerosis

Point counting represents a convenient and efficient technique for estimating the area of transects through multiple sclerosis (MS) lesions on magnetic resonance (MR) images obtained for sections through the brain. When sectioning has been performed according to the Cavalieri method, unbiased estimates of the total volume of MR-visible MS plaques can be obtained with a precision of 3–5% in 5–10 min.     

Responses of Dairy Cows to Exogenous Bovine Growth Hormone Administered During Early Lactation

Bovine growth hormone was administered to eight Holstein cows during two 10-d periods (commencing 20 and 60 d postpartum). Treatments involved daily subcutaneous injection of excipient (control) or of 50 IU growth hormone. Feed intake and yields of milk and milk components were measured during the last 5 d of treatment. In the first injection period, milk yield of treated cows was 6% greater than that of controls (36.2 kg/d). Fat content of milk was increased by treatment with the result that fat yield was elevated by 25%. Protein content was not altered. In the second period, milk yield increased by 12%, but composition of milk was not changed. As a result, milk energy secretion was increased by a similar amount in each treatment period (4.4 versus 4.5 Mcal/d). Feed intake was not changed in either period. During early lactation the ability of cows to increase milk yield in response to exogenous growth hormone may be limited by the supply of glucose for lactose synthesis rather than by ability to mobilize body reserves in support of lactation.     

Ion implantation doping of OMCVD grown GaN

Doping by ion implantation using Si, O, Mg, and Ca has been studied in single crystal semi-insulating and n-type GaN grown on a-sapphire substrates. The n-and p-type dopants used in this study are Si and O; Mg and Ca, respectively. Room temperature activation of Si and O donors has been achieved after 1150°C annealing for 120 s. The activation of Mg and Ca acceptors is too low to measure at both room temperature and 300°C. Using higher doses to achieve a measurable p-type conduction increases the amount of damage created by the implantation. Rutherford back scattering measurements on this material indicate that the damage is still present even after the maximum possible heat treatment. Secondary ion mass spectrometry measurements have indicated a redistribution in the measured profiles of Mg due to annealing.     

Polymerase chain reaction amplification analyses of clonality in T-cell malignancy including peripheral T-cell lymphoma

Clonality in T-cell malignancy was investigated using T-cell receptor (TcR) V beta 1-20 family primers and polymerase chain reaction amplification (PCR) of cDNA prepared from tissue biopsies and blood involved with tumour. Secondary PCR amplification of the VDJ joints of primary PCR products was performed to distinguish clonal from polyclonal products, and clonal V beta gene products were confirmed by direct PCR sequencing in the majority of cases. In eight T-cell malignancies including T-cell acute lymphoblastic leukaemia (T-ALL) and T-cell chronic lymphocytic leukaemia (T-CLL) shown to be clonal by Southern blot analysis, one or two primary PCR products were identified and shown to be clonal. In five cases of peripheral T-cell lymphoma (PTCL) all V beta 1-20 families were identified after primary PCR amplification, and clonal products were identified in two cases after secondary amplification; TcR V beta clonal families could not be demonstrated in the remaining three cases. These data were in agreement with previous Southern blot analysis of these cases, and confirmed the presence of reactive T cells in PTCL as well as providing further evidence for the genotypic heterogeneity of this entity. In the remaining case, a blood lymphocytosis, primary PCR amplification produced predominant TcR V beta 6 and V beta 12 family products, of which the V beta 6 family proved clonal after secondary PCR amplification. There was no evidence for overrepresentation of TCR V beta families by the tumour populations in this study, furthermore the data confirm the involvement of reactive cells in T-cell malignancy and the genetic heterogeneity of PTCL.