Mini Nutritional Assessment Short-Form (MNA-SF) Predicts Institutionalisation in an Intermediate Post-Acute Care Setting.

OBJECTIVES: The aim of the study was to investigate whether nutritional status as measured by the Mini Nutritional Assessment Short-Form (MNA-SF) predicts level of care of the discharge destination after post-acute care in a nursing home. DESIGN: Cohort study. SETTING: Post-acute intermediate care nursing home, 19-bed unit with increased multidisciplinary staff. PARTICIPANTS: Patients ≥70 years in post-acute care (N=900) following an acute admission to the hospital. MEASUREMENTS: The predictive value of nutritional status, as measured by the MNA-SF, on discharge destination was analysed by means of a multinomial logistic regression model with the MNA-SF as the independent variable, discharge destination as the dependent variable and age and living situation as co-variates. The participants were grouped into three categories according to their discharge destination: home, other institution or readmitted to the hospital. RESULTS: A higher score on the MNA-SF significantly predicted a reduced risk of discharge to institution (adjusted OR=0.90 (95% CI=0.84;0.97), p=0.003). CONCLUSION: Nutritional status, as measured by the MNA-SF, predicted discharge destination from an intermediate care nursing home following acute hospitalisation. The findings underscore the value of nutritional assessment as part of overall clinical assessment and care planning prior to discharge. The findings may also imply that interventions to improve nutritional status could increase patients' ability to return home, as opposed to further institutionalisation, after hospitalisation.

Three Different Outcomes in Older Community-dwelling Patients Receiving Intermediate Care in Nursing Home after Acute Hospitalization.

OBJECTIVES: To evaluate the recovery and outcome of older communitydwelling patients admitted to intermediate care (IC) in nursing homes after acute hospitalization, and to compare patients who were able and unable to return directly to their own homes. DESIGN: Prospective, observational, cohort study conducted between June 2011 and 2014. SETTING: A 19- bed IC unit in a nursing home with increased multidisciplinary staffing. PARTICIPANTS: A total of 961 community-dwelling patients, ≥70 years of age, considered to have a rehabilitation potential and no major cognitive impairment or delirium, transferred from internal medicine, cardiac, pulmonary and orthopaedic hospital departments. MEASUREMENTS: Demographic data, clinical information, comprehensive geriatric assessment (CGA), discharge destination and length of stay. Residence status and mortality 1 month, 2 months, 3 months, and 6 months after discharge from the hospital. RESULTS: The trajectory of recovery was divided into 3 groups: 1) Rapid recovery, able to return home after median 14 days in IC (n=785, 82%); 2) Slow recovery, requiring additional transfer to other nursing home after IC, but still able to return home within 2 months (n=106, 11%). 3) Poor recovery, requiring transfer to other nursing home after IC and still in a nursing home or dead at 2 months (n=66, 7%). Significant different clinical characteristics were demonstrated between the patients in the 3 groups. After 6 months, the recovery of patients with rapid or slow recovery was similar, 87% were living at home, compared to only 20% of the patients with poor recovery. In multiple logistic regression analysis, slow or poor recovery was significantly associated with low scores on the Barthel index and orthopaedic admission diagnosis. CONCLUSIONS: Although the majority of patients selected for treatment in the IC unit were able to recover and return home, a group of patients needed extra time, up to 2 months, to recover and another group had a poor chance of recovering and returning home. Different caring pathways for different patient groups may be considered in the PAC setting.

Second malignancies after treatment of Hodgkin's disease: the influence of treatment, follow-up time, and age.

PURPOSE: In the period 1968 through 1988, The Norwegian Radium Hospital (NRH) treated an unselected population of 1,152 patients with Hodgkin's disease (HD) that comprised more older patients (mean age, 43 years) than most other institutions. We considered it important to evaluate these patients for development of second cancers (SCs). PATIENTS AND METHODS: The Norwegian Cancer Registry identified previously untreated patients with HD treated at NRH who had developed a SC more than 1 year after diagnosis of HD. The relative risk ratio (RR) (observed/expected cases) and the cumulative risk were calculated. RESULTS: Sixty-eight patients had developed a SC, including nine acute nonlymphocytic leukemias (ANLLs), eight non-Hodgkin's lymphomas (NHLs), and 51 solid tumors, including 11 lung cancers. The RR of SC and leukemia was 1.86 (95% confidence interval [CI], 1.4 to 2.4) and 24.3 (95% CI, 11.1 to 46.2), respectively. The RR of SC was highest in younger patients (< 41 years, RR = 3.8). No significant association between splenectomy and development of ANLL was found. The influence of treatment and follow-up time on the development of SC agrees with data from other large cancer institutions. CONCLUSION: (1) The low RR of developing a SC in this study is probably due to the number of older patients included, who have a lower RR of developing a SC due to less aggressive treatment, shorter follow-up time, and higher incidence of cancer in the expected background population. (2) The low RR and cumulative risk of developing ANLL may be due to the limited use of extensive chemotherapy (CT) in our hospital in the earlier years.

Flow cytometric measurement of apoptosis and necrosis in cryopreserved PBPC concentrates from patients with malignant diseases.

The number of viable precursor cells actually reinfused into patients after high-dose chemotherapy is one of the most clinically important variables determining graft success or failure. A modified, previously described flow cytometric method based on annexin V staining was therefore applied to assess the degree of apoptosis and necrosis in cryopreserved PBPC concentrates from patients with malignant diseases. Twenty-two samples of unmanipulated cryopreserved PBPC concentrates were analyzed by flow cytometry. The samples were triple-stained with anti-CD34 PE, annexin V-FITC and actinomycin D, which enabled the separation of viable, early apoptotic and late apoptotic/necrotic CD34(+) precursor cells. Apotosis and necrosis were also measured in the total cell population of the concentrates. Eighty-one percent (range 49-97) of the CD34(+) cells were viable, while 7% (range 1-15) were early apoptotic and 12% (range 2-36) were late apoptotic/necrotic after freeze/thaw. There was no difference in apoptosis and necrosis in CD34(+) cells harvested from mildly pretreated patients with multiple myeloma and heavily pre-treated patients with non-Hodgkin's lymphoma. Apoptosis and necrosis were higher in the total mature cell population of the concentrates. Thirty-two percent (range 7-69) of the cells were apoptotic and 33% (range 12-60) were necrotic. We conclude that flow cytometric analysis of annexinV/actinomycin D binding in PBPC concentrates is a simple technique that can give additional information of the viability status of the cells post thaw. The present study confirms the relative robustness of human CD34(+) precursor cells concerning the freeze/thaw procedure, which are carried out in daily clinical practice.

Circadian cell kinetics in humans. Aspects related to cancer chemotherapy.

A prerequisite for the investigation of cell kinetics, and especially circadian cell kinetics, has been the development from the 1950s and onwards of several methods for studying kinetic parameters in different mammalian tissues. A large number of such studies have subsequently taken place in the rodent, mostly as non-circadian experiments, but also a large number of studies have now documented on circadian proliferative rhythms in many different murine tissues. Results from cytokinetic studies in the human have also accumulated through the years. Of special interest has been the demonstration of temporal variations in rapidly proliferating tissues studies as the bone marrow and gut mucosa relative to cytotoxic anticancer therapy. Analyses of proliferation in human tumours have also indicated rhythms in malignant solid tumours. Thus, these studies have demonstrated that there exist rhythms in bone marrow and gut cytokinetics which increases the likelihood that certain times of day will be less toxic for the administration of cytotoxic drugs. Furthermore, optimizing anticancer therapy according to a circadian schedule may also increase the tumour cell death rat, due both to an indirect dose-escalating effect and a direct increased tumour effect.

Large-volume leukapheresis yields more viable CD34+ cells and colony-forming units than normal-volume leukapheresis, especially in patients who mobilize low numbers of CD34+ cells.

BACKGROUND: Large-volume leukapheresis (LVL) differs from normal-volume leukapheresis (NVL) by increased blood flow and altered anticoagulation regimen. LVL is now regarded as a safe procedure for collection of peripheral blood progenitor cells (PBPCs), but it is not known whether the procedure will alter CD34+ cell quality or will be useful for patients who mobilize few CD34+ cells into peripheral blood. STUDY DESIGN AND METHODS: The results from 82 LVL and 125 NVL (4.0-5.3 and 2.7-3.5 times the patients' blood volumes processed, respectively) were retrospectively analyzed in altogether 112 consecutive patients with malignant diseases. RESULTS: The LVL yielded significantly more CD34+ cells (4.2 x 10(6) vs. 3.1 x 10(6)/kg, p = 0.006, all patients; and 1.8 x 10(6) vs. 1.3 x 10(6)/kg, p = 0.004, bad mobilizers) and significantly higher colony-forming units (77 x 10(4) vs. 33 x 10(4)/kg; all patients and 33 x 10(4) vs. 20 x 10(4)/kg, p < 0.001, both groups). Significantly fewer leukapheresis procedures were required to obtain 2 x 10(6) CD34+ cells per kg (one vs. two, p = 0.001, all patients; and two vs. three, p = 0.009, bad mobilizers). No significant differences in CD34+ cell viability and time to hematologic recovery were observed between the patients who received PBPCs harvested by NVL and LVL. CONCLUSION: Although a median platelet loss of 36 percent can be expected, LVL can be recommended as the standard apheresis method for PBPC collections in patients with malignant diseases. LVL is particularly useful in patients who mobilize a low number of CD34+ cells into the peripheral blood.

Long-term morbidity after curative radiotherapy for carcinoma of the bladder. A retrospective study.

48 patients with muscle invasive bladder cancer were asked by a mailed questionnaire concerning post-radiotherapy functional status, urological and intestinal complaints. All patients had received curative pelvic irradiation (greater than or equal to 1750 reu) nine to 133 months prior to the investigation. Eleven patients had recurrent or persistent disease and four (36%) of these patients had severe hematuria. The urinary morbidity of the 37 patients without evidence of disease varied between 24 and 38%, but only 3 to 8% of these patients experienced severe disturbances. Four patients (9%) had to undergo bowl resection due to severe radiation damage in the intestines. In conclusion, slight morbidity after curative radiation treatment is to be expected in about 30% of patients surviving more than six months. Severe complications may be expected in 10% of the patients. Serious complaints from the urologic tract, especially hematuria, indicate recurrent or persistent disease rather than a late radiation effect.

Current concepts for chronochemotherapy of cancer.

In this article, a survey on the concepts and scientific basis for applying chemotherapy against malignant tumors on a circadian schedule is given. The idea is to give the cytostatic drugs at times of the day when optimal effect on the tumor is achieved, but at the same time causing minimal toxic side effects. Following a brief description of the complexity of cancer tissue, some aspects of the present status of cancer chemotherapy in general are reviewed. Applications of chronobiology in cancer treatment are then surveyed together with possibilities to increase cytostatic doses and reduce side effects. When optimal tumor cell kill is achieved, the next step is to address the circadian aspects of normal organs, including the proliferative behavior of tissues with rapid cell renewal. Finally, the question of how regulatory mechanisms responsible for normal circadian rhythms can be interfered with is addressed. Cancer chronochemotherapy today combined with modern infusional technology is a promising field for improving cancer treatment in general and reducing side effects and is expected to make important progress in the near future.

No differences in colony formation of peripheral blood stem cells frozen with 5% or 10% dimethyl sulfoxide.

High-dose chemotherapy with autologous stem cell rescue usually requires cryopreservation of the cells. For several years, 10% dimethyl sulfoxide (DMSO) has been used as the standard cryoprotectant. Because DMSO infusion can lead to toxic clinical complications in a dose-related manner, we wanted to evaluate if reduction to 5% DMSO would be possible. We have compared colony formation in the myeloid, erythropoietic, and megakaryocyte lineages in peripheral blood progenitor cell (PBPC) samples cryopreserved in parallel with 5% and 10% DMSO. Twenty-seven PBPC samples from patients with malignant diseases were investigated after 3 months of cryopreservation in liquid N(2), and samples from 14 of these patients were investigated after 1 year. A significantly higher colony formation was demonstrated for colony-forming units-erythrocyte (CFU-E) and CFU-granulocyte, erythrocyte, macrophage, megakaryocyte (GEMM) both at 3 months and at 1 year in the 5% samples. For CFU-granulocyte-macrophage (GM) and CFU-megakaryocyte (Mk) no significant difference was demonstrated neither at 3 months nor at 1 year in samples frozen with 5% and 10% DMSO. Also, there was a statistically significant correlation between the CFU-total and CFU-Mk-total, indicating that the CFU-total might be used as an evaluation of megakaryocyte progenitors. Viability testing with the Trypan Blue exclusion test showed that cells cryopreserved in 5% DMSO had significantly higher viability than the cells cryopreserved in 10% DMSO. We conclude that 5% DMSO is at least as good for cryopreservation of small-volume PBPC samples as the conventional 10% DMSO, and our results suggest that the possibility of using 5% DMSO for cryopreservation of autologous PBPC grafts should be further investigated in clinical studies.

Autologous transplantation: the viable transplanted CD34+ cell dose measured post-thaw does not predict engraftment kinetics better than the total CD34+ cell dose measured pre-freeze in patients that receive more than 2x10(6) CD34+ cells/kg.

BACKGROUND: The aim of the study was to investigate whether the number of viable CD34+ cells in cryopreserved PBPC autografts is a better predictor of engraftment than the total CD34+ cell number determined before freezing. METHODS: A total of 119 patients was treated with autotransplantation for various malignant disorders during the period 1996-2002. All patients were reinfused with at least 2x10(6)/kg total CD34 cells analyzed before programmed freezing in 10% DMSO. The total CD34 cell number determined before freezing was compared with the number of viable cells determined after cryopreservation for 51 of these patients. The number of viable cells was determined by a flow cytometric analysis including triple staining with anti-CD34, anti-CD45 and the viability marker 7-actinomycin D (7-AAD). RESULTS: Simple linear regression analyses showed that both the total transplanted CD34 cell dose measured before freezing and the viable CD34 cell dose determined after cryopreservation were significantly correlated with neutrophil and platelet engraftment. In a multiple regression model the prediction of engraftment was not improved when the transplanted viable CD34 cell dose was included as a variable in addition to the total CD34 cell dose measured immediately after collection. DISCUSSION: Routine estimation of viable CD34 cells after cryopreservation of PBPC autografts is not necessary as long as the total CD34 cell dose is determined before freezing and the patients are reinfused with at least 2x10(6) cells/kg body weight.

Peripheral blood progenitor cell grafts contain high levels of platelet-secreted mediators.

BACKGROUND: The cytokine network in peripheral blood progenitor cell (PBPC) grafts may affect hematopoietic reconstitution or the risk of postransplant relapse of malignant disorders through effects on normal progenitor cells or contaminating malignant cells. Whether thrombopoietin (TPO), SCF, and platelet-secreted mediators are parts of this network was investigated. STUDY DESIGN AND METHODS: Peripheral blood and PBPC plasma samples were collected consecutively from patients with malignant disorders who underwent PBPC harvest. Blood samples were collected immediately before and after apheresis. Patients underwent mobilization by chemotherapy plus G-CSF, except for one patient who received only G-CSF. Plasma levels were also determined for healthy controls. RESULTS: PBPC grafts had greater levels of platelet-secreted platelet factor 4 (PF4), beta-thromboglobulin, and platelet-derived growth factor isoform AB, as compared with venous levels in patients and controls. Although platelet and PF4 levels in autografts were significantly correlated, the graft:blood ratio was higher for PF4 than for platelets. In both the patients' blood and the autografts, TPO levels were increased from the levels in normal controls. Blood and graft levels of SCF were within the normal range. CONCLUSION: The cytokine network of PBPC autografts includes increased levels of TPO and several platelet-derived mediators.

Autologous stem cell transplantation as post-remission therapy in adult acute myelogenous leukemia: does platelet contamination of peripheral blood mobilized stem cell grafts influence the risk of leukemia relapse?

Conventional chemotherapy of acute myelogenous leukemia (AML) results in an overall long-term disease-free survival of less than 50%, but for selected subsets of younger patients the prognosis can be improved by allogeneic stem cell transplantation. The use of autologous stem cell transplantation is now investigated as an alternative to allotransplantation due to its lower risk of serious complications. However, autotransplantation is associated with a relatively high risk of post-transplant AML relapse that can be derived from contaminating leukemia cells in the autograft. Peripheral blood mobilized stem cell (PBSC) grafts usually contain a higher number of platelets. The degree of platelet contamination is determined by the peripheral blood platelet count at the time of harvesting, and the platelets become activated and release soluble mediators during the ex vivo handling of PBSC grafts. Many of these platelet-derived mediators can bind to specific receptors expressed by AML blasts, and the platelet contamination may then alter AML blast survival and thereby influence the risk of post-transplant leukemia relapse. Therefore, we conclude that the platelet contamination of autologous stem cell grafts is possibly of clinical importance, but the effect of this nonstandardized parameter is difficult to predict in individual patients because the number of graft-contaminating platelets, the degree of platelet activation, and the effects of platelet-derived mediators on AML blasts differ between patients.

[High dose chemotherapy with autologous stem cell support in cancer patients]. / Høydosebehandling med autolog stamcellestøtte hos kreftpasienter.

INTRODUCTION: In 1996 it was decided that high-dose chemotherapy with peripheral stem cell support should be offered by all five university hospitals in Norway. MATERIAL AND METHODS: We report on the first 49 patients from the western part of Norway treated with this modality at Haukeland University Hospital in the 1996-98 period. RESULTS: All patients had a total of > 2-10(6) CD34 positive cells/kg collected before high-dose chemotherapy. To achieve this critical stem cell dose, five patients had to have three or more stem cell collections; four of them had to be mobilised several times. Poor stem cell mobilisation was mostly marked in patients with soft tissue sarcoma and testicular cancer, but was also observed in a few heavily pre-treated patients with non-Hodgkins lymphoma. With the exception of one lymphoma patient who developed a rapid bone marrow relapse, all patients had satisfactory sign of bone marrow regeneration after reinfusion of the stem cells. This also applied to the poor mobilisors. No treatment-related deaths have occurred. Four to 38 months after high-dose therapy, 33% of patients with multiple myeloma and 52% of patients with malignant lymphoma were alive and in complete remission. Three of the four patients with soft tissue sarcoma relapsed 3-7 months after high-dose chemotherapy.

Circadian variation in serum cortisol and circulating neutrophils are markers for circadian variation of bone marrow proliferation in cancer patients.

Serum cortisol, circulating white blood cells and DNA cell cycle distribution in bone marrow cells were measured during daytime (11.00) and at midnight (24.00) over single 24-hour periods in 15 cancer patients. The neutrophils and fraction of bone marrow cells in S-phase showed the same circadian variation as cortisol with higher values in daytime as compared to midnight in 11 patients with a normal cortisol rhythm (p < 0.05). The lymphocytes, eosinophils and basophils all had significantly higher values at midnight as compared to daytime. There were significant correlations between cortisol and neutrophils, lymphocytes, eosinophils and basophils. The correlation between neutrophils and fractions of bone marrow cells in S-phase and S + G2/M-phase were highly significant (r = 0.74, p = 0.0001 and r = 0.72, p = 0.0001, respectively). In 8 of 13 patients (61.5%) without bone marrow infiltration both cortisol and neutrophils showed identical circadian variation as bone marrow cells in S-phase and S + G2/M-phase. Furthermore, for the total series a significant correlation between S-phase, cortisol and neutrophils was found by multiple regression analysis (p < 0.0001). These findings strengthen the possibility of using the circadian variation in cortisol and neutrophils as marker rhythms for circadian variation in bone marrow proliferation, thus allowing optimization of cytotoxic therapy and individualization of chronotherapy.

Bone marrow examination in Hodgkin's disease.

Bone marrow aspiration and biopsy was performed as part of routine staging in 425 patients with primary Hodgkin's disease. Only seven patients were found to have bone marrow disease by biopsy and only four by aspiration. All these patients had B symptoms and stage III or IV before bone marrow examination. Bone marrow infiltration did not influence treatment decision and there was no association between bone marrow findings and outcome of the disease.

DNA cell cycle distribution and glutathione (GSH) content according to circadian stage in bone marrow of cancer patients.

DNA cell cycle distribution and glutathione (GSH) content in bone marrow were measured both at daytime and midnight over single 24 h periods in 15 cancer patients. Between patients the S-phase demonstrated a difference from lowest to highest value of 700%, whereas the corresponding difference for the G2/M-phase was nearly 900%. The mean GSH content measured in the bone marrow at the two timepoints was 2.24 +/- 0.21 nmol mg-1 protein, range 0.91-4.19 nmol mg-1 protein. A statistically significant higher fraction of cells in S-phase and G2/M-phase was found at daytime as compared to midnight when excluding the four patients with an abnormal circadian variation in cortisol. No significant temporal variation in total bone marrow GSH content was found, although a weak correlation between S-phase and GSH content was demonstrated (r = 0.42; P less than 0.05). This correlation was strengthened when not including the six patients with an abnormal cortisol pattern (4) and bone marrow infiltration (2) (r = 0.66; P = 0.005). Cells in S-phase demonstrated a positive correlation with cells in G2/M-phase (r = 0.64; P less than 0.0001). A negative correlation was found between GSH content and age (r = 0.53; P less than 0.005). Finally, a statistically significant positive correlation was demonstrated between cortisol and both S-phase and G2/M-phase (r = 0.57; P less than 0.001 and r = 0.38; P less than 0.05, respectively). The present study suggests a possibility of optimising cancer therapy and use of hematopoietic growth factors by determining individual average values and circadian stage dependent variation in bone marrow DNA cell cycle distribution. Furthermore, GSH content in bone marrow may predict this tissue's sensitivity to cytotoxic agents.

Circadian cell cycle variations of erythro- and myelopoiesis in humans.

By use of a multiparameter flow cytometric method with specific surface markers, circadian (24-h) variations in cell cycle distribution have been studied in 19 healthy male volunteers by sampling bone marrow (BM) every 4-5 h during 24-h periods. Admixture of peripheral blood during the sampling was specifically adjusted for, and the fractions of cells in DNA synthetic phase were measured for different hemopoietic cell lineages. Significant circadian variations in DNA S-phase were demonstrated both in myelo- and erythropoiesis of the human BM, with 75% (myeloid) and 80% (erythroid) of the volunteers showing highest activity (values) of DNA S-phase during the day and lowest activity (values) between midnight and 04:00 h. A temporal relationship in the circadian variation of S-phase and G2/M-phase was demonstrated between the myeloid and erythroid cell lineages. The highest fractions of S-phase cells were found in erythropoiesis, while the highest circadian stage dependent variation was found in myelopoiesis. The existence of a similar phasing in DNA synthetic activity for myelopoietic and erythropoietic cells in the human bone marrow indicates that the circadian rhythmicity of hemopoiesis may be caused by a common regulatory mechanism. These findings may be relevant with regard to optimizing the use of cytotoxic drugs and hemopoietic growth factors by taking into consideration the intrinsic (endogenous) circadian variation in proliferative activity of human BM subpopulations.

Variation in cell yield and proliferative activity of positive selected human CD34+ bone marrow cells along the circadian time scale.

Variations in cell yield and proliferative activity of human bone marrow (BM) progenitor cells were determined with flow cytometry along the 24-h (circadian) time scale. Equal volumes of BM were aspirated every 5 h, altogether 5 times in 5 healthy men. An average 6-fold higher yield of positive selected CD34+ cells occurred in each subject when BM was aspirated during the daytime and late afternoon, while a lower yield occurred during the night. Using all CD34+ cell yield data normalized to percentage of mean, a significant time-effect was found by ANOVA (p = 0.02) and a significant circadian rhythm was detected by the least-squares fit of a 24 h cosine (p = 0.02). The 95% confidence limits of the acrophase (time of highest values) were computed to be at midday between 10:24 and 14:48 h. A highly significant correlation (p = 0.001) was found between proliferation of positive selected CD34+ cells and the more mature myeloid precursor cells from the same BM aspirates, suggesting a common temporal pattern along the circadian time scale. However, no correlation was demonstrated between proliferation and cell yield of CD34+ selected cells, suggesting that mechanisms other than variation in proliferation may cause the circadian rhythm in stem cell yield. These circadian variations in stem cell yield and proliferation suggest that proper timing within 24 h may potentially be important regarding outcome from progenitor cell harvesting and treatment with haematopoietic growth factors.

Insulin-like growth factor-1 (IGF-1) has a costimulatory effect on proliferation of committed progenitors derived from human umbilical cord CD34+ cells.

The effects of insulin-like growth factor-1 (IGF-1) on highly enriched human umbilical cord CD34+ cells were investigated in vitro. CD34+ cells were cultured in serum-free medium containing stem cell factor (SCF), GM-CSF, and interleukin-3 (IL-3). Culture of CD34+ cells for one week in the presence of these cytokines resulted in a dose-dependent increase in total cell number. Addition of G-CSF together with SCF+IL-3+GM-CSF increased the proliferation of myelopoietic cells as determined by the number of cells expressing the myelomonocytic marker CD64 and the granulocytic marker CD15 without significantly altering the number of CD34+ cells in the cultures. In the presence of G-CSF, IGF-1 induced a dose-dependent increase in the total cell number and a moderate but significant increase in the percentages of CD15+, CD64+ cells with sustained CD34+ cell proliferation. We conclude that IGF-1 can enhance the in vitro proliferation of committed progenitor cells derived from umbilical cord CD34+ cells.

Flow cytometric measurement of DNA S-phase in human bone marrow cells: correcting for peripheral blood contamination.

The relationship between bone marrow (BM) cells with S-phase DNA content and the amount of peripheral blood contamination estimated as percentage lymphocytes+monocytes (L+MO) present in BM samples has been investigated in a total of 136 BM aspirates and biopsy expellates from 35 hematologically healthy individuals. A significant negative correlation was demonstrated between total, erythroid and myeloid BM cells in S-phase and the percentage of L+MO in the aspirates (r=0.84, 0.57 and 0.49, respectively; p<0.0001). Based on the equation of the slope of the regression line, a correction formula adjusting the measured value of BM cells in S-phase to varying amounts of L+MO percentage has been worked out for the total and erythroid BM cells. In contrast, highly proliferating myelomonocytic cells and CD34+ cells did not show any significant correlation between cells in S-phase and percentage L+MO, indicating that peripheral blood contamination of BM aspirates estimates the degree of peripheral blood contamination, as well as make possible a correct estimation of the DNA synthesis of several BM populations. The method is especially applicable when frequent BM sampling is required.