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We have recently shown that more than 90% of long-term culture initiating cells (LTC-IC) mobilized in the peripheral blood (PB) of normal individuals express HLA-DR and CD38 antigens and can sustain hematopoiesis for only 5 weeks. However, 10% of LTC-IC in mobilized PB are CD34+ HLA-DR- and CD34+ CD38- and can sustain hematopoiesis for at least 8 weeks. We now examine the ex vivo expansion potential of CD34+ HLA-DR+ cells (rich in mature LTC-IC) and CD34+ HLA-DR- cells (rich in primitive LTC-IC) in granulocyte colony-stimulating factor (G-CSF) mobilized PB progenitor cells (PBPC). Cells were cultured in contact with M2-10B4 cells (contact) or in transwells above M2-10B4 (noncontact) without and with interleukin-3 (IL-3) and macrophage inflammatory protein (MIP-1alpha) for 2 and 5 weeks. Progeny were evaluated for the presence of colony-forming cells (CFC) and LTC-IC. When CD34+ HLA-DR+ PB cells were cultured in contact cultures without cytokines, a threefold expansion of CFC was seen at 2 weeks, but an 80% decrease in CFC was seen at week 5. Further, the recovery of LTC-IC at week 2 was only 17% and 1% at week 5. This confirms our previous observation that although CD34+ HLA-DR+ mobilized PB cells can initiate long-term cultures, they are relatively mature and cannot sustain long-term hematopoiesis. In contrast, when CD34+ HLA-DR- mobilized PB cells were cultured in contact cultures without cytokines, CFC expansion persisted until week 5 and 49% and 11% of LTC-IC were recovered at week 2 and 5, respectively. As we have shown for steady state bone marrow (BM) progenitors, recovery of LTC-IC was threefold higher when CD34+ HLA-DR- PBPC were cultured in noncontact rather than contact cultures, and improved further when IL-3 and MIP-1alpha were added to noncontact cultures (96 +/- 2% maintained at week 5). We conclude that although G-CSF mobilizes a large population of "mature" CD34+ HLA-DR+ LTC-IC with a limited proliferative capacity, primitive CD34+ HLA-DR- LTC-IC present in mobilized PB have similar characteristics as LTC-IC from steady state BM: (1) they can be maintained in noncontact cultures containing IL-3 and MIP-1alpha for at least 5 weeks; (2) they are subject to the same proliferation inhibitory influences of contact with stroma. Since the absolute number of primitive LTC-IC (week 8 LTC-IC) per mL of G-CSF mobilized PB is similar to that per mL of steady state BM, these studies further confirm that G-CSF mobilized PBPC may have similar long-term repopulating abilities as steady state BM.

Although more than 3 million head of dairy cows enter the food supply chain in the U.S. every year, research on this topic remains limited and scarce. Meat production from dairy cows is a significant component of beef production, accounting for almost 10% of U.S. commercial beef production. Thus, the purpose of this review is to demonstrate the importance of dairy cows as a beef source, and to provide an overview on topics from farm to meat product—culling, marketing, transportation, welfare, body composition and its relationship with lactation particularities, carcass characteristics, meat quality, and traceability. Current scientific evidence has shown that culling a dairy cow at an appropriate time has beneficial effects on cow welfare and, consequently, cow value. During marketing the dairy cow is visually evaluated for health and factors associated with its expected carcass value; thus, marketing a well-conditioned cow will ensure that the animal is fit for transportation and provides high carcass yield. However, limitations such as low body condition score, lameness or mobility problems, and visual defects remain persistent. Even though beef harvest plants accommodate cows in all body composition states, the current carcass grade system does not reflect the mature cow industry needs. Therefore, improvement of the grading system could maximize carcass utilization and increase cow carcass value by recognizing subprimal cuts that could be merchandised as whole muscle cuts. Lastly, implementation of a traceability system would unify information from the farm to harvest assisting the industry in making further advancements. [ABSTRACT FROM AUTHOR]

The relaxation of N2(X¹Σg+) molecules in a background gas composed of N(4Su) atoms and free electrons is studied by using an ideal isochoric and isothermic chemical reactor. A rovibrational state-to-state model is developed to study energy transfer process induced by free electron and atomic collisions. The required cross sections and the corresponding rate coefficients are taken from two well-known kinetic databases: NASA Ames kinetic mechanism for the description of the N2(X¹Σg+) - N(4Su) processes and the Phys4Entry database for the electron driven processes, N2(X¹Σg+) -e-. The evolution of the population densities of each individual rovibrational level is explicitly determined via the numerical solution of the master equation for temperatures ranging from 10000 to 30 000 K. It was found that the distribution of the rovibrational energy levels of N2(X¹Σg+) is strongly influenced by the electron driven collisional processes, which promote the excitation of the low lying vibrational levels. The macroscopic vibrational energy relaxation is governed by the molecule-atom collisions, when free electrons, initially cold are relaxing to the final heat-bath temperature. Thus, the main role of the free electrons is to ensure the equilibration of vibrational and free electron excitation, thus validating the existence of the local equilibrium TV-Te. However, if electrons and heavy particles are assumed to be in equilibrium at the heat bath temperature, electron driven processes dominate the vibrational relaxation. Finally, we have assessed the validity of the Landau-Teller model for the description of the inelastic energy transfer between molecules and free electrons. In the case of free-electron temperatures lower than 10 000 K, Landau-Teller relaxation model gives an accurate description of the vibrational relaxation, while at higher temperatures the error in the predictions can be significant and the model should not be used. [ABSTRACT FROM AUTHOR]

Cull dairy cows contribute almost 10% of national beef production in the United States. However, different factors throughout the life of dairy cows affect their weight and overall body condition as well as carcass traits, and consequently affect their market price. Therefore, the objectives of this study were: (1) to assess relationships between price ratio and carcass merit of cull dairy cows sold through several sites of an auction market and (2) to investigate the effect of animal life history events and live weight on sale barn price (BP) and price ratio (as a measure of relative price), as an indicator of carcass merit. Data from 4 dairy operations included 3,602 cull dairy cow records during the period of 2015 to 2019. Life history events data were collected from each dairy operation through Dairy Comp software; live weight and price were obtained periodically from the auction market, and the carcass data were provided by a local packing plant. Cow price in dollars per unit of live weight ($/cwt) and price ratio were the 2 outcome variables used in the analyses. Price ratio was created aiming to remove seasonality effects from BP (BP divided by the national average price for its respective month and year of sale). The association between price ratio and carcass merit traits was investigated using canonical correlation analysis, and the effect of life history events on both BP and price ratio was inferred using a multiple linear regression technique. More than 70% of the cows were culled in the first 3 lactations, with an average live weight of 701.5 kg, carcass weight of 325 kg, and dressing percentage of 46.3%. On average, cull cows were sold at $57.0/cwt during the period considered. The canonical correlation between price ratio and carcass merit traits was 0.76, indicating that price ratio reflected carcass merit of cull cows. Later lactations led to lower BP compared with cows culled during the first 2 lactations. Injury, and leg and feet problems negatively affected BP. Productive variables demonstrated that the greater milk production might lead to lower cow prices. A large variation between farms was also noted. In conclusion, price ratio was a good indicator of carcass merit of cull cows, and life history events significantly affected sale BP and carcass merit of cull cows sold through auction markets. [ABSTRACT FROM AUTHOR]

Abstract: Founder effects introduce stochasticity in the genetic structure of species at the regional scale. To the extent that founder effects are important, they will result in a reduced signature of space, time, and environmental variation in landscape genetic data. We studied the metapopulation genetic structure of recently founded populations of the microcrustacean Daphnia sinensis in 10 Ethiopian water reservoirs. We used three different approaches to estimate the number of effective founders and applied them to two independent genetic marker sets in an effort to investigate the role of founder effects and to estimate effective size of the founding population. Estimates of founding sizes rarely exceeded eight individuals but were most often limited to less than four individuals. No associations of genetic identities, gene frequencies, measures of genetic diversity, or differentiation with environmental and spatial variables were found. Age and size of the reservoirs were not correlated with genetic diversity measures or number of founders in these reservoirs. These findings indicate that neither strong selection, nor dispersal limitation are responsible for the observed pattern of genetic variation. Our results suggest a regional population structure that is strongly impacted by founder events, reflecting colonization by just a few founders per waterbody, and not noticeably influenced by subsequent dispersal and gene flow. Our results show that rapid colonization of empty habitats and fast population growth by a handful of founders can result in strong founder effects, even in relatively large habitats that are likely regularly reached by new immigrants. [ABSTRACT FROM AUTHOR]

Human impact often leads to reduced population sizes, and populations exposed to anthropogenic stress may suffer reduced evolutionary potential because of loss of adaptive genetic variation and higher risk of inbreeding depression (ID). Here, we exposed naive and carbaryl-selected inbred and outbred subpopulations of Daphnia magna to the pesticide carbaryl shortly after birth, and monitored acute (day 1-4) and post-exposure mortality (day 5-second clutch). Overall, acute mortality was lower than post-exposure mortality, indicating predominantly long-term costs of carbaryl exposure. Surprisingly, we found no indication for ID with respect to mortality upon carbaryl exposure. This may be due to more effective purging of deleterious alleles under standard conditions in the more homozygous inbreds as compared to the more heterozygous outbreds. Alternatively, homozygous pesticide resistance alleles in inbreds may render the inbreds an advantage compared to outbreds, where such resistance alleles would more likely occur heterozygotically. Additionally, we found that the capacity to further reduce mortality in response to carbaryl selection tended to be reduced in inbreds compared to outbreds. Our results thus suggest that inbred lineages may cope equally well as outbreds with pesticide stress. [ABSTRACT FROM AUTHOR]

New habitats are an interesting tool to monitor colonization processes in the light of changes in genetic structure and evolutionary potential of populations. Cladocerans are cyclical parthenogens that offer the possibility to track genetic changes during the colonization process due to the alternation of sexual and asexual phases. The locally called 'malladas' are shallow and temporary peridunal ponds in Eastern Spain that were silted in the 1960s and have been recently dug to their original basin. We here describe the colonization process in two of these newly restored ponds by the cladoceran Daphnia magna during their first 3 years. This colonization process followed three critical steps: population founding by the initial colonists, arrival of secondary immigrants and consolidation of the population structure. We found a low number of genotype colonists followed by the establishment of secondary immigrants and a slight decrease in gene flow with time. We discuss the importance of dispersal, founder effects, natural selection, inbreeding depression and genetic drift during the colonization process in the light of the most recent literature. [ABSTRACT FROM AUTHOR]

Cyclical parthenogens are a valuable system in which to empirically test theoretical predictions as to the genetic consequences of sexual reproduction in natural populations, particularly if the frequency of sexual relative to asexual reproduction can be quantified. In this study, we used a series of lake populations of the cyclical parthenogen, Daphnia pulicaria, that vary consistently in their investment in sexual reproduction, to address the questions of whether the ecological variation in investment in sex is detectable at the genetic level, and if so, whether the genetic patterns seen are consistent with theoretical predictions. We show that there is variation in the genetic structure of these populations in a manner consistent with their investment in sexual reproduction. Populations engaging in a high frequency of sex were in Hardy-Weinberg and gametic phase equilibrium, and showed little genotypic differentiation across sampled years. In contrast, populations with a lower frequency of sex deviated widely from equilibrium, had reduced multilocus clonal diversity, and showed significant temporal genotypic deviation. [ABSTRACT FROM AUTHOR]

Relative effects of Beef Quality Assurance (BQA)-related defects in market beef and dairy cows and bulls on selling price at auction was evaluated during 2008. The presence and severity of 23 BQArelated traits were determined during sales in Idaho, California, and Utah. Overall, 18,949 unique lots consisting of 23,479 animals were assessed during 125 dairy sales and 79 beef sales. Mean sale price ± SD (per 45.5 kg) for market beef cows, beef bulls, dairy cows, and dairy bulls was $45.15 ± 9.42, $56.30 ± 9.21, $42.23 ± 12.26, and $55.10 + 9.07, respectively. When combined, all recorded traits explained 36% of the variation in selling price in beef cows, 35% in beef bulls, 61% in dairy cows, and 56% in dairy bulls. Premiums and discounts were determined in comparison with a "par" or "base" animal. Compared with a base BCS 5 beef cow (on a 9-point beef scale), BCS 1 to 4 cows were discounted (P < 0.0001), whereas premiums (P < 0.05) were estimated for BCS 6 to 8. Compared with a base BCS 3.0 dairy cow (on a 5-point dairy scale), more body condition resulted in a premium (P 0.001), whereas a less-than-desirable BCS of 2.0 or 2.5 was discounted (P < 0.0001). Emaciated or near-emaciated cows (beef BCS 1 or 2; dairy BCS 1.0 or 1.5) were discounted (P < 0.0001). Compared with base cows weighing 545 to 635 kg, lighter BW beef cows were discounted (P < 0.0001), whereas heavier beef cows received (P < 0.05) a premium. Compared with a base dairy cow weighing 636 to 727 kg, lighter BW cows were discounted (P < 0.0001), whereas heavier cows (727 to 909 kg) received a premium (P < 0.01). Beef and dairy cows with any evidence of lameness were discounted (P < 0.0001). Presence of ocular neoplasia in the precancerous stage discounted (P = 0.05) beef cows and discounted (P < 0.01) dairy cows, whereas at the cancerous stage, it discounted (P < 0.0001) all cows. Hide color influenced (P < 0.0001) selling price in beef cattle but had no effect (P = 0.17) in dairy cows. Animals that were visibly sick were discounted (P < 0.0001). Results suggest that improving BCS and BW, which producers can do at the farm or ranch level, positively affects sale price. Furthermore, animals that are visibly sick or have a defect associated with a possible antibiotic risk will be discounted. Ultimately, animals with minor quality defects should be sold in a timely manner before the defect advances and the discount increases. [ABSTRACT FROM AUTHOR]

A survey was conducted to quantify incidence of Beef Quality Assurance (BQA)-related defects in market beef and dairy cows and bulls selling at auction during 2 seasons in 2008. Twenty-three BQArelated traits were evaluated by 9 trained personnel during sales at 10 livestock auction markets in Idaho (n = 5; beef and dairy), California, (n = 4; dairy only), and Utah (n = 1; beef and dairy). Overall, 18,949 unique lots (8,213 beef cow, 1,036 beef bulls, 9,177 dairy cows, and 523 dairy bulls,) consisting of 23,479 animals (9,299 beef cows, 1,091 beef bulls, 12,429 dairy cows, and 660 dairy bulls) were evaluated during 125 sales (64 spring, 61 fall) for dairy and 79 sales (40 spring, 39 fall) for beef. The majority of market beef cows and bulls (60.9 and 71.3%, respectively) were predominantly black-hided, and the Holstein hide pattern was observed in 95.4 and 93.6% of market dairy cows and bulls, respectively. Market cattle weighed 548 ± 103.6 kg (beef cows), 751 ± 176.1 kg (beef bulls), 658 ± 129.7 kg (dairy cows), and 731 + 150.8 kg (dairy bulls). Most beef cows (79.6%) weighed 455 to 726 kg, and most beef bulls (73.8%) weighed 545 to 954 kg, respectively. Among market beef cattle, 16.0% of cows and 14.5% of bulls weighed less than 455 and 545 kg, respectively, and 63.7% of dairy cows and 81.5% of dairy bulls weighed 545 to 817 kg or 545 to 954 kg, respectively. However, 19.5% of dairy cows and 13.1% of dairy bulls weighed less than 545 kg. Mean BCS for beef cattle (9-point scale) was 4.7 + 1.2 (cows) and 5.3 ± 0.9 (bulls), and for dairy cattle (5-point scale) was 2.6 ± 0.8 (cows) and 2.9 + 0.6 (bulls). Some 16.5% of beef cows and 4.1% of beef bulls had a BCS of 1 to 3, whereas 34.8% of dairy cows and 10.4% of dairy bulls had a BCS of 2 or less. Emaciation (beef BCS = 1, dairy BCS = 1.0) or near-emaciation (beef BCS = 2, dairy BCS = 1.5) was observed in 13.3% of dairy cows and 3.9% of beef cows. Among beef cattle, 15.1% of cows and 15.4% of bulls were considered lame. In contrast, 44.7% of dairy cows and 26.1% of dairy bulls were lame. Ocular neoplasia (cancer eye) was observed in only 0.6% of beef cows, 0.3% of beef bulls, 0.3% of dairy cows, and 0.0% of dairy bulls. However, among animals with ocular neoplasia, it was cancerous in 34.4% of beef bulls, 48.0% of dairy cows, and 73.3% of beef cows. In conclusion, numerous quality defects are present in market beef and dairy cattle selling at auction in the Western United States, which could influence their value at auction. [ABSTRACT FROM AUTHOR]

The ubiquity of sexual reproduction is an evolutionary puzzle because asexuality should have major reproductive advantages. Theoretically, transitions to asexuality should confer substantial benefits in population growth and lead to rapid displacement of all sexual ancestors. So far, there have been few rigorous tests of one of the most basic assumptions of the paradox of sex: that asexuals are competitively superior to sexuals immediately after their origin. Here I examine the fitness consequences of very recent transitions to obligate parthenogenesis in the cyclical parthenogenetic rotifer Brachionus calyciflorus. This experimental system differs from previous animal models, since obligate parthenogens were derived from the same maternal genotype as cyclical parthenogens. Obligate parthenogens had similar fitness compared with cyclical parthenogens in terms of the intrinsic rate of increase (calculated from life tables). However, population growth of cyclical parthenogens was predicted to be much lower: sexual female offspring do not contribute to immediate population growth in Brachionus, since they produce either males or diapausing eggs. Hence, if cyclical parthenogens constantly produce a high proportion of sexual offspring, there is a cost of sex, and obligate parthenogens can invade. This prediction was confirmed in laboratory competition experiments. [ABSTRACT FROM AUTHOR]

The reproductive mode of facultative parthenogens allows recessive mutations that accumulate during the asexual phase to be unmasked following sexual reproduction. Longer periods of asexual reproduction should increase the accumulation of deleterious mutations within individuals, reduce population-level genetic diversity via competition and increase the probability of mating among close relatives. Having documented that the investment in sexual reproduction differs among populations and clones of Daphnia pulicaria, we ask if this variation is predictive of the level of inbreeding depression across populations. In four lake populations that vary in sex investment, we raised multiple families (mother, field-produced daughter, laboratory-produced daughter) on high food and estimated the fitness reduction in both sexually produced offspring relative to the maternal genotype. Inbred individuals had lower fitness than their field-produced siblings. The magnitude of fitness reduction in inbred offspring increased as population-level investment in sex decreased. However, there was less of a fitness reduction following sex in the field-produced daughters, suggesting that many field-collected mothers were involved in outcross mating. [ABSTRACT FROM AUTHOR]

In populations of a cyclical parthenogen, the diversity of clonal lineages, derived from sexually produced eggs, declines during the parthenogenetic phase. Even though Daphnia magna populations from small ponds may harbour millions of individuals, we show that observed clonal and allelic diversity in populations from such small water bodies are lower than in populations from larger water bodies. Populations from small water bodies also show significant fluctuations in allele frequencies among years and a stronger among-population genetic differentiation than populations inhabiting larger water bodies. Persistent founder effects can only explain part of these results. Our data link the population genetic structure of cyclical parthenogens to the size of the habitat and suggest that genetic drift is a more prominent feature of populations inhabiting small water bodies than previously thought.Heredity (2007) 98, 419–426. doi:10.1038/sj.hdy.6800958; published online 28 March 2007 [ABSTRACT FROM AUTHOR]

The Keynote Paper reviews and highlights over 40 years of research on solutions for steady-state and transient thermal constriction and spreading resistances, and thermomechanical models for contact, gap and joint resistances of joints formed by conforming rough surfaces, nonconforming smooth surfaces, and nonconforming rough surfaces. Microgap and macrogap thermal resistance and conductance models are reviewed, and important relations and correlation equations are presented. Contact microhardness, determined by Vickers indenters, are correlated and incorporated into the contact model for conforming rough surfaces. Microhardness parameters are correlated with Brinell hardness values. Elastoplastic contact models for joints formed by smooth sphere-smooth flat and conforming rough surfaces are presented. A simple thermomechanical model for microgaps occupied by oil, grease, grease filled with solid particles, and phase change materials such as paraffins is reviewed, and good agreement with recently published data is noted. [ABSTRACT FROM AUTHOR]

Summary:The progenitor content of autologous peripheral blood progenitor and stem cell collections is a major determinant of prompt hematopoietic recovery following autologous stem cell transplantation. We analyzed unstimulated bone marrow (BM) and peripheral blood (PB) apheresis products in comparison to those collected following G-CSF or GM-CSF stimulation. We quantitated their committed (CFU-GM) and primitive (long-term culture-initiating cells, LTC-IC) progenitors in relation to hematologic recovery in 63 patients undergoing autografting for lymphoid malignancies. G-CSF, but not GM-CSF, substantially enriched the committed progenitor content (2.5-3.6-fold) of both PB and BM grafts. G-CSF also enriched the LTC-IC content of BM and PB compared to control grafts. GM-CSF augmented (11.5-fold) the LTC-IC content of stimulated BM, but not GM-CSF-mobilized PB. Neutrophil recovery was substantially quicker in recipients of BM or PB mobilized with G-CSF or GM-CSF. In contrast, red cell and platelet recovery was accelerated in recipients of GM-CSF-stimulated BM (but not PB) and G-CSF-stimulated PB (but not BM). No direct correlation between progenitor dose and hematopoietic recovery for neutrophils, platelets or red cells was observed. Cytokine stimulation can augment the committed and more primitive multilineage progenitor content of BM and PB grafts, to a differing extent. The uncertain relationship with multilineage myeloid recovery emphasizes the limitations in using clonogenic progenitor analyses to assess the adequacy of an autologous graft prior to transplantation.Bone Marrow Transplantation (2003) 32, 575-580. doi:10.1038/sj.bmt.1704237 [ABSTRACT FROM AUTHOR]

Summary. Ex vivo expansion of primitive human haematopoietic stem cells (HSC) is clinically relevant for stem cell transplantation and gene therapy. Here, we demonstrate the selective expansion of CD34+ CD38– cells from purified CD34+ cells upon stimulation with Flt3-ligand, stem cell factor and thrombopoietin. Over a 100-fold (range 80 to 128-fold) expansion of CD34+ CD38– cells was observed with bone marrow and cord blood (CB). The expanded CD34+ CD38– cells remained negative for lineage-specific markers and could be induced to differentiate into granulocytes, monocytes, megakaryocytes, erythrocytes, and T and B-lymphocytes in vitro . Lineage differentiation assays with single CD34+ CD38– cells showed no loss of multilineage potential of expanded cells after ex vivo culture. We also demonstrated that the increase in frequency of CD34+ CD38– cells was not as a result of the downregulation of CD38 expression during the culture. Quantitative analysis showed that the number of 6 week cobblestone area forming cells (CAFCwk6 ), a measure of proliferating HSC, in cytokine-stimulated CD34+ cells were increased by 20-fold. Expanded CD34+ CD38– cells could be transduced efficiently with retroviruses encoding the low affinity nerve growth factor receptor (LNGFR) marker gene (17% to 44%, mean 27%), resulting in long-lasting expression of retroviral-encoded genes in progeny HSC and differentiated progenitors. We conclude that the combination Flt3-ligand (FL), stem cell factor and thrombopoietin (TPO) induced strong ex vivo proliferation of CD34+ CD38– cells and that the absolute number of expanded cells with stem cell activity increased substantially in this population. [ABSTRACT FROM AUTHOR]

Transplantation of progenitor cells which have been mobilised into the bloodstream (PBPC) following the administration of G-CSF results in more rapid neutrophil recovery than transplantation of bone marrow (BM). The reasons for the accelerated neutrophil engraftment are not clear, but would be explained by increased self-replication of myeloid progenitor cells (CFU-GM). We have used a CFU-GM replating assay to investigate myeloid progenitor self-replication, and quantification of subcolony formation during erythroid burst formation to quantify erythroid progenitor self-renewal. Secondary colony formation by CFU-GM, grown from PBPC and then replated was increased compared with secondary colony formation by BM CFU-GM (P = 0.0001); erythroid subcolony formation was not altered. There was no difference between the replating abilities of PBPC CFU-GM derived from allogeneic donors (normal individuals) and autologous donors (patients with malignant disease) although differences were found between subgroups of autologous donors. The increased replication of PBPC could not be accounted for by a reduction in progenitor cell apoptosis; PBPC CFU-GM contained slightly fewer apoptotic CD34+ cells than BM CFU-GM. The increased replication by PBPC CFU-GM was reversible because it declined when CFU-GM colonies were passaged through three sequential CFU-GM replating cycles. This decline in self-replication was more rapid than the decline seen in replated BM CFU-GM. The self-replication of PBPC CFU-GM, and subcolony formation by BFU-E could be further enhanced by exposure to cytokines in vitro. We conclude that mobilisation alters the replication kinetics of myeloid, but not of erythroid, progenitor cells, that mobilisation-induced events are of limited duration and that in vitro exposure to cytokines may modify PBPC progenitor cell kinetics. Bone Marrow Transplantation (2001) 27, 241–248. [ABSTRACT FROM AUTHOR]

The use of mobilized peripheral blood (PB) stem cells for autologous transplantation initially generated much enthusiasm because of enhanced engraftment in comparison to marrow stem cells and avoidance of general anesthesia for the donor. Its application to the allogeneic setting seemed inevitable. For obvious ethical reasons, allogeneic donors are mobilized with cytokines only, mainly granulocyte colony-stimulating factor (G-CSF). Results from preliminary studies suggest that in comparison to standard bone marrow transplants, outcomes such as engraftment, host-versus-graft reaction, graft-versus-host disease, graft-versus-leukemia and immunological reconstitution may be different. Surprisingly, G-CSF, previously recognized as a late acting lineage-specific factor for neutrophil production, not only disrupts homeostasis between stem cells and their microenvironment, but also induces significant quantitative and qualitative changes in the accessory cell compartment, affecting lymphocytes, monocytes, natural killer, dendritic, and stromal cells. Furthermore, mobilization of huge numbers of non-professional antigen presenting cells (CD34+ stem cells) amplifies the tolerizing potential of PB stem cell grafts. Thus, G-CSF mobilization provides PB transplants with different immunobiologic properties in comparison to standard bone marrow grafts. Whether these immunobiologic differences will lead to better transplant outcomes remains to be shown through much awaited results of large randomized clinical trials. Bone Marrow Transplantation (2000) 26, 1–16. [ABSTRACT FROM AUTHOR]

CD34+-enriched peripheral blood stem cells (PBSC) are increasingly being used as an autograft in patients with multiple myeloma (MM). The rationale for the use of the CD34+-enriched fraction in MM is the ability to obtain a graft with a significant reduction of contamination by plasma cells. However, the effect of such a manipulation on the proliferating potential of the engrafted cells is not known. We wished to study, as part of a randomized trial comparing the outcome in MM patients transplanted with either CD34+- enriched cells or unfractionated PBSC, the primitive hematopoietic cell content of the autografts using long-term culture initiating cell (LTC-IC) assays in 7 MM patients. In 3 patients CD34+cell-enriched fraction was compared to unfractionated PBSC whereas in the remaining 4 patients the LTC-IC assay was performed on total PBSC. The mean percentage of CD34+ cells of the CD34+ selected fraction in three patients was 82% (range 71%-96%) whereas the same percentage in PBSC varied from 0.6% to 10% in 4 patients (mean: 4.2%). Out of three patients transplanted with CD34+ cell fraction, two patients were found to have a very similar LTC-IC generating potential in their CD34+ versus PBSC fractions as this was assessed by the clonogenic cell output at week+5 per 104 CD34+ cells initiating the culture (PBSC: 92 and 168 and CD34+ fraction: 102 and 161, respectively) whereas one patient had a slightly different values (PBSC: 51 and CD34+ fraction: 103). When the PBSC fraction was compared in all 7 patients, the LTC-IC generation potential was very heterogenous, varying from 1.4 to 168. To determine if the selection procedure influences the numbers of LTC-IC’s in both fractions, we have performed limiting dilution assays to determine both the frequency of distribution of hematopoietic colonies and the frequency of LTC-IC’s in two patients. The frequency of distribution of hematopoietic colonies was linear in both CD34+ and PBSC fractions as was the frequency of LTC-IC when the corrections were made with regard to the CD34+ cell-content of the cultures (1/20). Our results indicate that the CD34+ selection procedure used in all three patients (Ceprate) is not deleterious for the generation of LTC-IC’s and these findings support the rationale for the use of this procedure in multiple myeloma for the purposes of tumor depletion. [ABSTRACT FROM AUTHOR]

We have studied a total of 188 patients with hematological malignancies, submitted to mobilization therapy with G-CSF associated or not with chemotherapy in order to: (1) establish the lower limit of circulating progenitor cells that allows the collection of 2 × 106CD34+ cells/kg by a single leukapheresis, utilizing the instrument set on standard parameters; (2) evaluate whether the number and quality of CD34+ cells collected remain stable during leukapheresis; and (3) collect a sufficient number of circulating CD34+ cells by a single procedure in patients in whom such an approach would have been insufficient to reach the target with the instrument set on standard parameters. The retrospective analysis conducted in 85 patients showed that 19 circulating CD34+ cells/μl represented the cut-off number capable of discriminating between patients who will require one or more apheresis to collect 2 × 106 CD34+ cells/kg. The validity of this value was prospectively confirmed in 70 subsequent patients. Based on in vitroresults that showed the stability in the number of CD34+ cells, the proportion of different CD34+ cell subpopulations and the clonogenic capacity of the stem cell compartment during leukapheresis both in the blood of the patients and in samples taken directly from the instrument, we have adapted the blood volume to be processed in 33 patients with <19 pb cd34+ cells/μl. Stem cell collection was monitored during the leukapheresis and the procedure was prolonged for a time period estimated to be sufficient to reach the target number of CD34+ cells with a single procedure. The median increment of the total blood volume processed, calculated from the volume set automatically by the instrument was 25.2%, with a median of 3.3-fold total blood volume processed. In all cases, a sufficient CD34+ cell collection was... [ABSTRACT FROM AUTHOR]

Imatinib-insensitive leukemia stem cells (LSCs) are believed to be responsible for resistance to BCR-ABL tyrosine kinase inhibitors and relapse of chronic myelogenous leukemia (CML). Identifying therapeutic targets to eradicate CML LSCs may be a strategy to cure CML. In the present study, we discovered a positive feedback loop between BCR-ABL and protein arginine methyltransferase 5 (PRMT5) in CML cells. Overexpression of PRMT5 was observed in human CML LSCs. Silencing PRMT5 with shRNA or blocking PRMT5 methyltransferase activity with the small-molecule inhibitor PJ-68 reduced survival, serial replating capacity, and long-term culture-initiating cells (LTC-ICs) in LSCs from CML patients. Further, PRMT5 knockdown or PJ-68 treatment dramatically prolonged survival in a murine model of retroviral BCR-ABL-driven CML and impaired the in vivo self- renewal capacity of transplanted CML LSCs. PJ-68 also inhibited long-term engraftment of human CML [CD34.sup.+] cells in immunodeficient mice. Moreover, inhibition of PRMT5 abrogated the Wnt/β-catenin pathway in CML [CD34.sup.+] cells by depleting dishevelled homolog 3 (DVL3). This study suggests that epigenetic methylation modification on histone protein arginine residues is a regulatory mechanism to control self-renewal of LSCs and indicates that PRMT5 may represent a potential therapeutic target against LSCs., Introduction Chronic myelogenous leukemia (CML) is a disease of hematopoietic stem cells (HSCs) malignantly transformed by the formation of the Philadelphia chromosome (i.e., BCR-ABL fusion gene) due to reciprocal chromosomal [...]