Treatment intensity and symptom burden in hospitalized adolescent and young adult hematopoietic cell transplant recipients at the end of life.

Adolescent and young adult (AYA) oncology patients experience many physical and psychological symptoms at the end of life (EOL); however, data on these experiences for AYA patients who have undergone hematopoietic cell transplantation (HCT) remains sparse. We sought to investigate the characteristics of AYA patients aged 15-25 years who received allogeneic HCT and subsequently died while inpatient at our institution between the years 2008 and 2014. A standardized data extraction tool was used to collect information about patient demographics, treatment and symptoms. We found that during this time frame, 34 AYA patients had received HCT and died while inpatient at our institution, 23 (68%) of whom died because of treatment-related complications. Compared with non-HCT AYA oncology patients (n=35), patients who received HCT (n=34) were more likely to have died in the intensive care unit (71% vs 23%, P<0 .0001) and to have received mechanical ventilation (68% vs 17%, P<0.0001) or hemodialysis (53% vs 0%, P<0.0001) in the last 30 days of life. These findings demonstrate that AYA patients who receive allogeneic HCT receive intensive EOL treatment, suggesting that these patients may benefit from early integration of expert interdisciplinary services to prospectively assess and manage distressing symptoms.

Rapid memory T-cell reconstitution recapitulating CD45RA-depleted haploidentical transplant graft content in patients with hematologic malignancies.

T-cell depletion of an HLA-haploidentical graft is often used to prevent GVHD, but the procedure may lead to increased graft failure, relapse and infections due to delayed immune recovery. We hypothesized that selective depletion of the CD45RA+ subset can effectively reduce GVHD through removal of naive T cells, while providing improved donor immune reconstitution through adoptive transfer of CD45RA- memory T cells. Herein, we present results from the first 17 patients with poor-prognosis hematologic malignancy, who received haploidentical donor transplantation with CD45RA-depleted progenitor cell grafts following a novel reduced intensity conditioning regimen without TBI or serotherapy. Extensive depletion of CD45RA+ T cells and B cells, with preservation of abundant memory T cells, was consistently achieved in all 17 products. Neutrophil engraftment (median day +10) and full donor chimerism (median day +11) was rapidly achieved post transplantation. Early T-cell reconstitution directly correlated with the CD45RA-depleted graft content. T-cell function recovered rapidly with broad TCR Vß spectra. There was no infection-related mortality in this heavily pretreated population, and no patient developed acute GVHD despite infusion of a median of >100 million per kilogram haploidentical T cells.

Risk-adapted donor lymphocyte infusion based on chimerism and donor source in pediatric leukemia.

Donor lymphocyte infusion (DLI) is commonly used to treat leukemia relapse following stem cell transplantation. In florid relapse, however, the efficacy of DLI is limited with substantial risk of severe graft-versus-host disease (GvHD). Here, we develop a novel risk-adapted strategy characterized by pre-emptive DLI initiated at the time of mixed chimerism, a small starting dose based on donor source, dose-escalation guided by real-time chimerism monitoring and withholding of DLI immediately in patients achieving full donor chimerism. A total of 178 DLIs were given to 38 patients with mixed chimerism; thereafter, 33 patients (86.8%) had donor chimerism successfully increased, including 30 (78.9%) who had chimerism fully converted back to 100% donor. Cumulative incidence of relapse was significantly lower (P=0.00004) and overall survival higher (P=0.0003) in patients with chimerism fully corrected as compared with those of patients whose chimerism remained mixed. Only 13.2% of the patients developed acute grade III-IV GvHD with no associated mortality. In conclusion, the risk-adapted DLI strategy is useful in minimizing the risk of childhood leukemia relapse, GvHD and death.

Effects of activating NK cell receptor expression and NK cell reconstitution on the outcomes of unrelated donor hematopoietic cell transplantation for hematologic malignancies.

Inhibitory NK cell receptors are recognized as important determinants of NK cell activity in hematopoietic cell transplantation (HCT). The role of activating receptors and their acquisition after HCT is less certain. Therefore, we comprehensively evaluated both inhibitory and activating receptors in 59 patients receiving unrelated donor HCT. NK cell numbers normalized quickly relative to B and T cells; however, the expression of both inhibitory and activating isoforms of killer immunoglobulin-like receptors (KIRs) was delayed. Most NK cells expressed an immature phenotype during the first 6 months post-HCT; however, we found high expression of activating NKp46 and NKp44 natural cytotoxicity receptors (NCRs), and cytotoxicity was preserved. Early reconstituting NK cells from unmanipulated grafts showed lower cytotoxicity than those from T-cell-depleted grafts. Differences in NK cell reconstitution had significant effects on clinical outcomes. Patients whose NK cells reconstituted earlier had better survival and lower relapse rates. The best survival group was recipients who possessed HLA-C2 but their donor lacked the cognate-activating KIR2DS1. Collectively, our data underscore the clinical relevance of reconstituting NK cells and their activating KIRs and NCRs. In addition to NK cell quantification and genotyping, comprehensive assessment of NK cell functions and phenotypes, including activating receptors, is essential.

Gene-specific changes in alpha-tubulin transcript accumulation in developing cotton fibers.

The fibers of cotton (Gossypium hirsutum) are single-cell trichomes that undergo rapid and synchronous elongation. Cortical microtubules provide spatial information necessary for the alignment of cellulose microfibrils that confine and regulate cell elongation. We used gene-specific probes to investigate alpha-tubulin transcript levels in elongating cotton fibers. Two discrete patterns of transcript accumulation were observed. Whereas transcripts of alpha-tubulin genes GhTua2/3 and GhTua4 increased in abundance from 10 to 20 d post anthesis (DPA), GhTua1 and GhTua5 transcripts were abundant only through to 14 DPA, and dropped significantly at 16 DPA with the onset of secondary wall synthesis. This is the first report, to our knowledge, of gene-specific changes in tubulin transcript levels during the development of a terminally differentiated plant cell. The decrease in abundance of GhTua1 and GhTua5 transcripts was correlated with pronounced changes in cell wall structure, suggesting that alpha-tubulin isoforms may be functionally distinct in elongating fiber cells. Although total alpha-tubulin transcript levels were much higher in fiber than several other tissues, including the hypocotyl and pollen, none of the alpha-tubulins was specific to fiber cells.

Testicular and epididymal function during the peripuberal period in Brahman bulls receiving various amounts of protein degradable in the rumen.

Thirty-nine Brahman bulls with an initial age and weight of 301.7 +/- 4.1 d and 202.7 +/- 4.7 kg, respectively, were randomly allocated to 1 of 2 dietary treatment groups within age, weight and sire in order to study the influence of source of protein and stage of peripuberal period on testicular and epididymal function. In the soybean meal treatment the amount of protein undegradable in the rumen averaged 47%, while it was 72% in the fish meal treatment. The supplements were isocaloric and isonitrogenous. Bulls were electroejaculated, and castrations were performed randomly in a predetermined order when the first ejaculate with the first motile sperm cells (Stage 1), 10 to 25 million (Stage 2), and 50 million or more sperm cells (Stage 3 - puberty) was obtained. Testicular and epididymal traits were analyzed for a single testicle and epididymis. Daily sperm production, daily sperm production per gram of testicular parenchyma, testicular weight and testicular parenchyma weight were not affected by treatment. Bulls receiving fish meal had heavier (P < 0.01) epididymis than soybean meal-fed bulls (6.6 +/- 1.0 vs 3.9 +/- 0.6 g) but similar (P > 0.05) epididymal sperm reserves. Daily sperm production (1 testicle) was 115.2 +/- 0.1, 447.4 +/- 0.1, 792.7 +/- 0.1 million sperm cells, and daily sperm production per gram of testicular parenchyma was 1.5 +/- 0.5, 3.2 +/- 0.6 and 6.4 +/- 0.6 million sperm cells for bulls at Stage 1, 2 and 3, respectively. Sire and amount of undegradable intake protein had significant (P < 0.05) affects on the distribution of epididymal sperm reserves, with soybean meal-fed bulls having the higher proportions of epididymal sperm reserves in the cauda epididymis.

Influence of undegraded intake protein supplementation on milk production, weight gain, and reproductive performance in postpartum Brahman cows.

Eighty first-calf Brahman heifers and 51 mature Brahman cows were allotted to one of three diets based on parity, sex of calf, and breed of calf sire (Angus, Brahman, or Tuli) to evaluate rumen undegraded intake protein's (UIP) influence on production characteristics and reproductive performance. Supplements contained either 38.1% (low), 56.3% (medium), or 75.6% (high) UIP. Supplements were given from d 7 to 119 after calving to dams grazing rye-ryegrass overseeded Coastal bermudagrass pastures and with access to Coastal bermudagrass hay. Dam and calf BW and dam body condition score were recorded on d 7, 35, 63, 91, and 119 after calving. Four-hour milk production was recorded on the above days for low (n = 18), medium (n = 19), and high (n = 18) UIP animals and on d 7 and 35 for the entire group. Blood for progesterone RIA was drawn weekly and on d 6, 8, 10, and 12 after an observed estrus. Medium UIP heifers produced more (P < .02) milk (1.18 +/- .07 kg/4 h) than high UIP heifers (.94 +/- .07 kg/4 h), but milk production in mature cows was not influenced by diet. Low UIP dams had lower (P < .04) first-service conception rates (29.2%) than medium (57.6%) and tended (P < .10) to have lower rates than high UIP dams (54.6%). Overall pregnancy rates tended (P < .10) to be higher in medium (61.5%) and high (56.4%) UIP groups than in the low (43.2%) UIP group. Supplementing UIP at the medium rate improved first-service conception rates and tended to improve pregnancy rates.

Effect of ruminally undegradable protein from fish meal on growth and reproduction of peripuberal Brahman bulls.

Thirty-nine Brahman bulls (301.7 +/- 4.1 d; 202.7 +/- 4.7 kg) were allotted to one of two treatments and fed soybean meal (SBM)- or fish meal (FIS)-based supplements and hay to examine the effects of source of protein on growth and reproductive development. The fish meal supplement had 72% ruminally undegradable protein (RUP) and the soybean meal supplement had 47% RUP. Bulls assigned to the FIS treatment had higher (P < .01) total weight gain (81.2 +/- 1.4 vs 71.2 +/- 2.2 kg), higher (P < .01) ADG (.97 +/- .02 vs .85 +/- .03 kg), and better (P < .05) feed:gain ratio (7.6 +/- .1 vs 8.6 +/- .1 feed/BW gain for FIS vs SBM, respectively). Age at first motile spermatozoa was not affected (P > .05) by source of protein (429.9 +/- 9.6 vs 427.2 +/- 9.5 d, for bulls receiving FIS or SBM supplements, respectively). Likewise, age at puberty (473.3 +/- 21.7 d vs 465.9 +/- 12.9 d for bulls receiving FIS and SBM supplements, respectively) was similar for both treatment groups. There were no differences between treatments in scrotal circumference at those stages. At puberty semen quality was similar for bulls receiving FIS or SBM treatments, and no differences existed in LH and testosterone concentrations between treatments. We conclude that fish meal supplement increased growth but did not alter reproductive parameters in Brahman bulls.

Changes in the Accumulation of [alpha]- and [beta]-Tubulin Isotypes during Cotton Fiber Development.

The expression of [alpha]- and [beta]-tubulin proteins in developing fibers and several other tissues of cotton (Gossypium hirsutum, cv Texas Marker 1) have been analyzed by immunoblots of one- and two-dimensional gels utilizing anti-tubulin antibodies as probes. As a percentage of total protein, fibers had greater amounts of tubulin than did hypocotyls, roots, leaves, or cotyledons. Both [alpha]- and [beta]-tubulin, having apparent molecular masses of approximately 50 kD and isoelectric points between pH 5 and pH 6, were resolved on a single two-dimensional gel. Under the conditions used, [alpha]-tubulin was less acidic in the isoelectric focusing dimension and migrated slightly faster in the sodium dodecyl sulfate dimension than did [beta]-tubulin. Nine [alpha]-tubulin isotypes that formed two distinct groups were identified on immunoblots of two-dimensional gels. The three most abundant [alpha]-tubulin isotypes were common to all tissues examined. Seven distinct [beta]-tubulin isotypes were also identified. Although their level of accumulation differed, four of the [beta]-tubulin isotypes were common to all tissues. Preferential accumulation of isotypes was more apparent in fibers than in the other tissues examined. Two [alpha]-tubulin isotypes and two [beta]-tubulin isotypes showed preferential accumulation in 10- and 20-d postanthesis fibers, respectively.