John Mandeli

Non-myeloablative conditioning and allogeneic transplantation for multiple myeloma†

In multiple myeloma (MM), allogeneic stem cell transplantation (alloHCT) carries a lower relapse risk than autologous transplantation but a greater transplant‐related mortality. Nonmyeloablative conditioning for allogeneic transplantation (NST) reduces transplant‐related toxicity. Results are encouraging when used during first remission in low‐risk patients, but less‐so in relapsed or refractory disease. This is a single‐center retrospective analysis of 20 previously treated MM patients who underwent NST from matched‐related or matched‐unrelated donors from 2000–2006. Median age was 52.7 years (37.2–68.0). Twenty‐five percent had advanced or high‐risk disease. Eleven still had active disease prior to NST. Conditioning was total body irradiation 200 cGy on a single fraction on day −5, followed by antithymocyte globulin (ATG) 1.5 mg/kg/day and fludarabine 30 mg/m2/day on days −4 to −2. All received immunosuppression, most commonly with oral mycofenylate mofetil and cyclosporine beginning on day −5. At day 100, 50% had achieved complete remission. Transplant‐related mortality was 25%. Median overall survival (OS) was 21.2 months (0.6–90+) and progression‐free survival (PFS) 6.6 months (0.6–90+). Both OS and PFS were 24% at 3 years. OS was significantly greater for patients with age <52 years (median 27 months vs. 7.9 months, P = 0.031), and there was a trend toward greater OS for those with β2 microglobulin <2.5 mg/l (median 27 months vs. 7.7 months, P = 0.08). Donor characteristics and Ig type had no significant effect on survival. These data suggest a benefit of NST in relapsed/refractory MM. Randomized trials must be performed to confirm and further qualify this benefit. Am. J. Hematol., 2010.

The effect of dihydrotestosterone and culture conditions on proliferation of the human prostatic cancer cell line LNCaP

Cell density, nutritional state, and serum factors modify the growth response of LNCaP human prostatic cancer cells to dihydrotestosterone. Evaluation of growth response to dihydrotestosterone requires logarithmic transformation of cell count or thymidine incorporation data. Under conditions of dose response, growth increases with cell density but no significant interaction of dihydrotestosterone with cell density was found under optimal culture conditions. The frequency of media change was a significant factor in modulating dose response. When cells from cultures maintained at different feeding periods were plated at different cell densities of (trypan blue) viable cells, significant effects of plating density on dihydrotestosterone response were found. Dihydrotestosterone protects cells under the adverse effects of media deprivation. Under the extreme adverse effects of serum deprivation, cells respond to dihydrotestosterone even under conditions of increasing cell loss. The effects of dihydrotestosterone on final cell density were significant. In the absence of serum, the elongated cells of LNCaP assume a round shape, but many remain adherent to the culture dish and can be restored to normal morphology by serum. A number of growth factors fail to restore normal morphology that was completely restored by a combination of fibronectin and dihydrotestosterone. We have not developed a practicable serum-free system for LNCaP.

Combined bone marrow and peripheral blood progenitor cell autografts for patients with poor mobilization

BACKGROUND AIMSnPeripheral blood progenitor cell (PBPC) autografts with low CD34(+) cell content provide inadequate platelet (Plt) and red blood cell (RBC) reconstitution. Repeat collection and bone marrow (BM) harvesting are used in this situation. Minimum cell contents for BM-PBPC combined grafts are undefined.nnnMETHODSnA retrospective analysis of 19 autologous stem cell transplants (ASCT) with combined BM-PBPC for poor initial PBPC collection was carried out. Mobilization was with filgrastim (10 microg/kg/day) alone for 5 days or after chemotherapy. BM was harvested if PBPC collections were CD34+<2.5 x 10(6)/kg.nnnRESULTSnThe median age was 55 years (range 19-74). The diagnoses were multiple myeloma (7), non-Hodgkins lymphoma (7), Hodgkins disease (4) and acute myeloid leukemia (1). The median cell content (CD34+/kg x 10(6)) was 1.1 (0.3-2.7) for BM, 1.2 (0.04-2.8) for PBPC and 2.2 (1.4-4.9) combined. Eight grafts contained <2.0 x 10(6) CD34+/kg (1.4-1.8). The median engraftment in days (range) was: absolute neutrophil count (ANC) > 500, 12 (9-39); Plt > 20 000, 25 (15-70); RBC transfusion independence, 17 (6-93). Six patients died of progressive disease (58-293 days post-ASCT), one of infection on day 141 and one of AML on day 11. All patients except one maintained ANC > 1000 without filgrastim support beyond day 19. One patient had cholecystitis and delayed graft failure on day 90. PBPC CD34+ content did not predict CD34+ BM content but correlated with ANC > 500 (r= - 0.64, P=0.003). BM and combined CD34+ and BM TNC/kg did not correlate with engraftment or outcomes. Combined CD34+/kg < or > = 2.0 x 10(6) produced similar engraftment and mortality.nnnCONCLUSIONSnAfter a failed PBPC collection, BM harvest is a reliable option for obtaining an adequate combined autograft. Combined BM-PBPC autografts with <2.0x10(6) CD34+/kg can produce satisfactory engraftment.

Serum of patients with prostatic cancer or benign prostatic hypertrophy contains nonpolar testosterone.

We have previously described a nonpolar form of radioimmunoassayable serum testosterone (NPT) not measured by available antitestosterone antibodies. It is detected by mild alkaline hydrolysis of the petroleum ether extract of serum and subsequent radioimmunoassay. The properties of NPT are consistent with that of a fatty acid ester of testosterone or dihydrotestosterone. The serum of young males contains 1 to 3 ng/ml NPT, but it is not detected in female serum. We measured serum testosterone and NPT levels in 36 men between 58 and 87 years of age. Seventeen subjects with advanced prostatic cancer (NPT 1.70 +/- 1.44 ng/ml) were compared with a control group consisting of six patients with benign prostatic hypertrophy (BPH) and 13 patients with no prostatic disease (NPT 0.72 +/- 0.46, P = 0.017). There was no significant difference between BPH patients and patients with no prostatic disease; the results were pooled. The concentration of NPT in prostatic cancer patients but not in controls was inversely correlated with that of testosterone. Immunoassayable testosterone was present in the serum of two orchiectomized patients and, therefore, cannot derive solely from the testes.

Progesterone/Estradiol Ratios Predict Outcome in In Vitro Fertilization

Despite increasing experience with in vitro fertilization (IVF), there are few endocrinological markers which can serve as a prospective predictor of IVF success. An initial investigation utilized periovulatory estradiol (E2) levels after administration of low dose human menopausal gonadotropin followed by a coasting period before giving human chorionic gonadotropin (hCG). The levels of estradiol studied were lower than those typically seen in many IVF programs today.1 Others have studied post-transfer progesterone/E2 ratios and suggested a predictive value.2 This report summarizes our efforts to further elucidate endocrinological markers that can be used to predict success or potential success in IVF.