A modified comorbidity index for hematopoietic cell transplantation.

A recent validation analysis at our center among allogeneic hematopoietic cell transplant (HCT) recipients did not find the HCT-specific comorbidity index (HCT-CI) to clearly segregate patient's transplant-related risk. We hypothesized that the discriminating and predictive power of the HCT-CI for mortality could be improved by eliminating the assignment of categorical weights to comorbidities and instead replacing them with hazard ratios (HR) from a Fine and Gray adjusted regression model. This approach allowed us to look carefully at each component of the comorbidity index. We developed the modified comorbidity index (MCI) using a cohort of 444 adult allogeneic HCT recipients using a pure multiplicative model. Compared with low-risk patients, the HR for non-relapse mortality (NRM) using the HCT-CI was 1.3 (95% confidence intervals, 0.7-2.4) for intermediate risk and 1.6 (0.9-2.8) for high-risk patients, and with the MCI was 1.6 (0.9-2.8) and 2.7 (1.5-5.0), respectively. In conclusion, we are introducing the MCI which may have higher discriminating and predictive power for overall survival and NRM. Validation of the HCT-CI and the MCI in larger and separate cohorts of HCT recipients is still needed.

Non-random inactivation of large common fragile site genes in different cancers.

The common fragile sites are regions of profound genomic instability found in all individuals. The full size of each region of instability ranges from under one megabase (Mb) to greater than 10 Mbs. At least half of the CFS regions have been found to span extremely large genes that spanned from 600 kb to greater than 2.0 Mbs. The large CFS genes are also very interesting from a cancer perspective as several of them, including FHIT and WWOX, have already demonstrated the capacity to function as tumor suppressor genes, both in vitro and in vivo. We estimate that there may be 40-50 large genes localized in CFS regions. The expression of a number of the large CFS genes has been previously shown to be lost in many different cancers and this is frequently associated with a worse clinical outcome for patients. To determine if there was selection for the inactivation of different large CFS genes in different cancers, we examined the expression of 13 of the 20 known large CFS genes: FHIT, WWOX, PARK2, GRID2, NBEA, DLG2, RORA isoforms 1 and 4, DAB1, CNTNAP2, DMD, IL1RAPL1, IMMP2L and LARGE in breast, ovarian, endometrial and brain cancers using real-time RT-PCR analysis. Each cancer had a distinct profile of different large CFS genes that were inactivated. Interestingly, in breast, ovarian and endometrial cancers there were some cancers that had inactivation of expression of none or only one of the tested genes, while in other specimens there was inactivation of multiple tested genes. Brain cancers had inactivation of many of the tested genes, a number of which function in normal neurological development. We find that there is no relationship between the frequency that any specific CFS is expressed and the frequency that the gene from that region is inactivated in different cancers. Instead, it appears that different cancers select for the inactivation of different large CFS genes.

DMD and IL1RAPL1: two large adjacent genes localized within a common fragile site (FRAXC) have reduced expression in cultured brain tumors.

Common fragile sites (CFSs) are large regions of profound genomic instability found in all individuals. Spanning the center of the two most frequently expressed CFS regions, FRA3B (3p14.3) and FRA16D (16q23.2), are the 1.5 Mb FHIT gene and the 1.0 Mb WWOX gene. These genes are frequently deleted and/or altered in many different cancers. Both FHIT and WWOX have been demonstrated to function as tumor suppressors, both in vitro and in vivo. A number of other large CFS genes have been identified and are also frequently inactivated in multiple cancers. Based on these data, several additional very large genes were tested to determine if they were derived from within CFS regions, but DCC and RAD51L1 were not. However, the 2.0 Mb DMD gene and its immediately distal neighbor, the 1.8 Mb IL1RAPL1 gene are CFS genes contained within the FRAXC CFS region (Xp21.2-->p21.1). They are abundantly expressed in normal brain but were dramatically underexpressed in every brain tumor cell line and xenograft (derived from an intracranial model of glioblastoma multiforme) examined. We studied the expression of eleven other large CFS genes in the same panel of brain tumor cell lines and xenografts and found reduced expression of multiple large CFS genes in these samples. In this report we show that there is selective loss of specific large CFS genes in different cancers that does not appear to be mediated by the relative instability within different CFS regions. Further, the inactivation of multiple large CFS genes in xenografts and brain tumor cell lines may help to explain why this type of cancer is highly aggressive and associated with a poor clinical outcome.

RORA, a large common fragile site gene, is involved in cellular stress response.

Common fragile sites (CFSs) are large genomic regions present in all individuals that are highly unstable and prone to breakage and rearrangement, especially in cancer cells with genomic instability. Eight of the 90 known CFSs have been precisely defined and five of these span genes that extend from 700 kb to over 1.5 Mb of genomic sequence. Although these genes reside within some of the most unstable chromosomal regions in the human genome, they are highly conserved evolutionarily. These genes are targets for large chromosomal deletions and rearrangements in cancer and are frequently inactivated in multiple tumor types. There is also an association between these genes and cellular responses to stress. Based upon the association between large genes and CFSs, we began to systematically test other large genes derived from chromosomal regions that were known to contain a CFS. In this study, we demonstrate that the 730 kb retinoic acid receptor-related orphan receptor alpha (RORA) gene is derived from the middle of the FRA15A (15q22.2) CFS. Although this gene is expressed in normal breast, prostate and ovarian epithelium, it is frequently inactivated in cancers that arise from these organs. RORA was previously shown to be involved in the cellular response to hypoxia and here we demonstrate changes in the amount of RORA message produced in cells exposed to a variety of different cellular stresses. Our results demonstrate that RORA is another very large CFS gene that is inactivated in multiple tumors. In addition, RORA appears to play a critical role in responses to cellular stress, lending further support to the idea that the large CFS genes function as part of a highly conserved stress response network that is uniquely susceptible to genomic instability in cancer cells.

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Nutrient-independent and nutrient-dependent factors stimulate protein synthesis in colostrum-fed newborn pigs.

We hypothesized that nonnutrient components, including growth factors, present in colostrum contribute to the stimulation of protein synthesis in colostrum-fed neonatal pigs. We studied neonatal pigs fed mature milk, colostrum, or a formula containing a macronutrient composition comparable to that of colostrum for 24 h. We measured the circulating concentrations of insulin, insulin-like growth factor I, glucose, and amino acids at intervals throughout the 24-h period, after which we measured in vivo protein synthesis using a flooding dose of [3H]phenylalanine. The rates of protein synthesis in several tissues measured after 24 h of feeding were greater than those we reported previously after 6 h of feeding. The acute (within 6 h) stimulation of protein synthesis in visceral and skeletal muscle tissues of neonatal pigs fed milk, colostrum, or formula was primarily influenced by nutrient intake and associated with rapid secretion of insulin. Indirect evidence suggests that intestinal absorption of ingested colostral insulin was minimal. However, the sustained increase in tissue protein synthesis between 6 and 24 h coincided with an increase in circulating insulin-like growth factor I. We found a novel, specific stimulation of skeletal muscle and jejunal protein synthesis in colostrum-fed pigs that can be attributed to some nonnutrient component of colostrum.

Interorder transfer of mycoplasmalike microorganisms between Drosophila paulistorum and Ephestia kühniella. II. Numbers of MLO and sterility.

A successful attempt was made to culture the mycoplasmalike microorganism causing semispecific hybrid male sterility in Drosophila paulistorum utilizing Ephestia kühniella as the intermediate host. Data gleaned from this passage indicates that the induction of sterility depends upon the quality not the quantity of infectious intracellular symbionts.