A phase II study of sirolimus, tacrolimus and rabbit anti-thymocyte globulin as GVHD prophylaxis after unrelated-donor PBSC transplant.

We report on a prospective phase II trial of 32 patients who underwent unrelated-donor hematopoietic cell transplantation, with a tacrolimus, sirolimus and rabbit anti-thymoctye globulin GVHD prophylactic regimen. The primary study endpoint was incidence of grades II-IV acute (aGVHD), with 80% power to detect a 30% decrease compared with institutional historical controls. Median age at transplant was 60 (19-71). In total, 23 patients (72%) received reduced-intensity conditioning, whereas the remainder received full-intensity regimens. Median follow-up for surviving patients was 35 months (range: 21-49). The cumulative incidence of aGVHD was 37.3%, and the 2-year cumulative incidence of chronic GVHD was 63%. We observed thrombotic microangiopathy in seven patients (21.8%), one of whom also developed sinusoidal obstructive syndrome (SOS). Four of the 32 patients (12.5%) failed to engraft, and 3 of these 4 died. As a result, enrollment to this trial was closed before the targeted accrual of 60 patients. Two-year OS was 65.5% and EFS was 61.3%. Two-year cumulative incidence of relapse was 12.5% and non-relapse mortality (NRM) was 15.6%. NRM and aGVHD rates were lower than historical rates. However, the unexpectedly high incidence of graft failure requires caution in the design of future studies with this regimen.

Multicenter analysis of novel and established variables associated with successful human islet isolation outcomes.

Islet transplantation is a promising therapy used to achieve glycometabolic control in a select subgroup of individuals with type I diabetes. However, features that characterize human islet isolation success prior to transplantation are not standardized and lack validation. We conducted a retrospective analysis of 806 isolation records from 14 pancreas-processing laboratories, considering variables from relevant studies in the last 15 years. The outcome was defined as post-purification islet equivalent count, dichotomized into yields > or =315 000 or < or =220 000. Univariate analysis showed that donor cause of death and use of hormonal medications negatively influenced outcome. Conversely, pancreata from heavier donors and those containing elevated levels of surface fat positively influence outcome, as did heavier pancreata and donors with normal amylase levels. Multivariable logistic regression analysis identified the positive impact on outcome of surgically intact pancreata and donors with normal liver function, and confirmed that younger donors, increased body mass index, shorter cold ischemia times, no administration of fluid/electrolyte medications, absence of organ edema, use of University of Wisconsin preservation solution and a fatty pancreas improves outcome. In conclusion, this multicenter analysis highlights the importance of carefully reviewing all donor, pancreas and processing parameters prior to isolation and transplantation.

Validation of methodologies for quantifying isolated human islets: an Islet Cell Resources study.

BACKGROUND: Quantification of islet mass is a crucial criterion for defining the quality of the islet product ensuring a potent islet transplant when used as a therapeutic intervention for select patients with type I diabetes. METHODS: This multi-center study involved all eight member institutions of the National Institutes of Health-supported Islet Cell Resources Consortium. The study was designed to validate the standard counting procedure for quantifying isolated, dithizone-stained human islets as a reliable methodology by ascertaining the accuracy, repeatability (intra-observer variability), and intermediate precision (inter-observer variability). The secondary aim of the study was to evaluate a new software-assisted digital image analysis method as a supplement for islet quantification. RESULTS: The study demonstrated the accuracy, repeatability and intermediate precision of the standard counting procedure for isolated human islets. This study also demonstrated that software-assisted digital image analysis as a supplemental method for islet quantification was more accurate and consistent than the standard manual counting method. CONCLUSIONS: Standard counting procedures for enumerating isolated stained human islets is a valid methodology, but computer-assisted digital image analysis assessment of islet mass has the added benefit of providing a permanent record of the isolated islet product being evaluated that improves quality assurance operations of current good manufacturing practice.

Measurements of temporal aspects of gait obtained with a multimemory stopwatch in persons with gait impairments.

STUDY DESIGN: Repeated measures of 14 temporal factors of gait obtained with a multimemory stopwatch from a variety of subjects with locomotor impairments. OBJECTIVES: To estimate the intratester and intertester reliability of 14 temporal factors of gait by using a multimemory stopwatch; to compare novice and expert clinicians at mastery of making these temporal measurements. BACKGROUND: Temporal gait measures are useful for describing the effectiveness of treatment interventions in patients with locomotor impairments. METHODS AND MEASURES: Eleven adult subjects (mean age, 48.4 years; SD, 5.7 years), 10 with locomotor impairments and 1 elderly adult, ambulated along a 6-m walkway 3 times at a self-selected walking speed. The subjects were videotaped from the side as they walked. Four physical therapists independently analyzed the videotapes on 2 occasions; 2 examiners were recent graduates, and 2 others had 23 years of clinical experience. Intraclass correlation coefficients were used to estimate intratester reliability. A component of variance analysis quantified the sources of variation. RESULTS: Intraclass correlation coefficients for each of the 14 variables varied from 0.88 to 0.98. The major contributor to variance was subject, followed by trial, error, and tester; the tester factor generally contributed less than 1% to the total variance. CONCLUSIONS: Reliable measurements of the temporal aspects of gait can be made by using a multimemory stopwatch and videotape in a clinical setting on patients with various locomotor problems. Our data suggest that measurements obtained by more experienced physical therapists were no more reliable than those made by recent graduates.

Myeloperoxidase and eosinophil cationic protein in serum and sputum during antibiotic treatment in cystic fibrosis patients with Pseudomonas aeruginosa infection.

I order to study the time-course of myeloperoxidase (MPO) and eosinophil cationic protein (ECP) as parameters for monitoring inflammation in cystic fibrosis (CF), we investigated ten patients during both a 14-day intravenous antibiotic treatment and a corresponding self control. Modified Shwachman-Kulczycki score improved significantly (p < 0.008), C-reactive protein (CRP) levels decreased significantly (p < 0.05) during antibiotic treatment, while in the control phase there were no significant differences. Lung function parameters did not change significantly during antibiotic treatment or control phase. Serum MPO concentration (p < 0.006) and peripheral blood neutrophil granulocyte counts (p < 0.04) decreased significantly during antibiotic treatment, but not during the control phase. Sentm ECP concentration showed a tendency to decrease during antibiotic treatment, but this failed to reach significance. In general, sputum concentrations of MPO and ECP Were 500- to 1000-fold higher than in serum. However, neither MPO nor ECP in sputum showed a significan variation over time during antibiotic treatment or control phase. From our data we conclude that: (1) measurements of MPO, neutrophils and CRP in peripheral blood do correlate with clinical parameters such as the modified Shwachman-Kulczycki score; (2) neutrophils and MPO seem to reflect inflammatory changes induced by antibiotic treatment; (3) eosinophils may play a role in CF by an enhanced 'releasability' and (4) Sputum measurements of mediators of inflammation cannot be recommended.

Honeybee nestmate recognition: Effects of queen fecal pheromones.

Previous work has shown that queen honeybees,Apis mellifera, produce waxy esters composed of 8-14 carbon acids and 6-14 carbon alcohols in their feces. We tested these esters for effects on nestmate recognition; 11 of the 12 esters tested significantly modified the recognition characteristics of worker honeybees. Pairwise tests showed that workers can discriminate between at least some pairs of queen esters and that workers can discriminate between a queen ester and hexadecane (another known nestmate recognition cue). These results suggest that a queen may use the esters to enable workers to recognize her or to scent-mark her colony.

Kin discrimination by worker honey bees in genetically mixed groups.

We tested the hypothesis that in a genetically mixed assemblage of worker honey bees, individual workers would behave differently toward unfamiliar sisters than toward unfamiliar nonsisters. Groups of worker honey bees of mixed genetic composition were assembled by collecting pupae from separate colonies and placing the worker bees together on eclosion. A total of 10 workers, 5 from each of two kin groups, were used to form each group. When the workers were 5 days old, a worker of one of the two kin groups was introduced into the mixed group. This worker had previously been held in a group of its sisters, without contact with queen or nonsister bees. The interactions with the introduced bee indicate that in a mixed kin group, individual workers learn the composite identity of the group and do not attack unfamiliar bees differentially on the basis of kinship. However, kinship does influence the total number of interactions in which an introduced bee engages when placed in a genetically mixed group; bees interacted significantly more often with sisters than with nonsisters. There was a trend for bees to be involved in more feeding interactions with sisters. This finding indicates an ability of a bee to learn and use its own cues. In mixed groups, each bee maintains its genotypically correlated identity; the bees' odors do not comingle into a "group" or "gestalt" odor. The significance of these results is discussed in light of the genetic structure of natural colonies of honey bees.