Fludarabine-based cytoreductive regimen and T-cell-depleted grafts from alternative donors for the treatment of high-risk patients with Fanconi anaemia

Eighteen consecutive patients aged 5·5–24 years with Fanconi anaemia and diagnoses of aplastic anaemia ( n  = 8), myelodysplastic syndrome ( n  = 4), acute myeloid leukaemia ( n  = 6), received allogeneic haematopoietic stem cell transplants from alternative donors. All patients had been transfused, 13 had previously been treated with androgens and 14 had a history of infection. Donors were related human leucocyte antigen (HLA) mismatched for eight patients, unrelated HLA mismatched for seven patients and unrelated HLA matched for three patients. Cytoreduction included single dose total body irradiation (450 cGy), fludarabine (150 mg/m²) and cyclophosphamide (40 mg/kg). Immunosuppression included antithymocyte globulin and tacrolimus. Grafts were granulocyte colony-stimulating factor-mobilized, CD34⁺ T-cell-depleted peripheral blood stem cells in 15 patients and T-cell-depleted marrows in three. All 18 patients engrafted with 100% donor chimaerism; only one patient developed graft-versus-host disease (GVHD). With a median follow-up of 4·2 years, 13/18 patients were alive, 12 of these were disease-free. Five-year overall survival and disease-free survival were 72·2% and 66·6% respectively. Immune reconstitution was achieved at approximately 6 months post-transplant for most patients. These are encouraging results of T-cell-depleted transplants from alternative donors using fludarabine-based cytoreduction in 18 high-risk patients with Fanconi anaemia, with no evidence of rejection and minimal GVHD.     

Regulation of translation initiation in mussels (Mytilus galloprovincialis,lam.) following contamination stress

The use of mussels (Mytilus sp.) as bioindicators of the aquatic environment is a valuable approach to monitor environmental contamination. Applying batteries of biomarkers is an essential prerequisite, since the complexity of environmental contaminants can induce in mussels a variety of structural and functional responses, which are not necessarily correlated. In an attempt to correlate translation responses to contamination stress, the sedimentation profiles of runoff ribosomes isolated from digestive gland cells of control or contaminated Mediterranean mussels (Mytilus galloprovincialis, Lam.) were examined and the efficiency of these ribosomes to accomplish protein synthesis was determined. While the major species of ribosomal material was 80S monomers, native 60S and 40S ribosomal subunits were also detected independently of the contamination level in the surrounding waters. However, concomitant with the increase of contamination stress, the level of 80S ribosomes was reduced in favor of free ribosomal subunits. In addition, ribosomes isolated from contaminated mussels and programmed with poly(U) were less efficient to enzymatically bind AcPhe-tRNA, compared with ribosomes from control samples. These results suggest that the contamination stress causes stoichiometric aberrations in the ribosomal particle pool and reduction of translation machinery capability to initate protein synthesis. Data support the notion that downregulation of translation is an important component of the cellular stress response and may be exploited as a biomarker of environmental contamination.     

Perforated Carcinoma of the Sigmoid Colon in an Incarcerated Inguinal Hernia: Report of a Case

Perforation of the large bowel due to benign or malignant disease in an inguinal hernia is very rare, but should be considered as a potential cause of strangulated hernias. A 79-year-old man with a 2-day history of scrotal swelling and pain in the left side associated with fever and chills was brought to our Emergency Department, where he was classified as American Society of Anesthesiologists IVE. A large left incarcerated scrotal hernia was diagnosed and surgical exploration was performed using local infiltration anesthesia. A standard oblique inguinal incision was made, revealing perforation of the sigmoid colon due to cancer. A 40-cm segmental resection of the sigmoid colon was done, and a double-barrel colostomy was made through the inguinal incision. This surgical strategy involving construction of a double-barrel colostomy through the inguinal hernia incision could be an alternative method of managing such critically ill patients.     

3'-axial CH2 OH substitution on glucopyranose does not increase glycogen phosphorylase inhibitory potency. QM/MM-PBSA calculations suggest why

Glycogen phosphorylase is a molecular target for the design of potential hypoglycemic agents. Structure‐based design pinpointed that the 3′‐position of glucopyranose equipped with a suitable group has the potential to form interactions with enzyme’s cofactor, pyridoxal 5′‐phosphate (PLP), thus enhancing the inhibitory potency. Hence, we have investigated the binding of two ligands, 1‐(β‐d ‐glucopyranosyl)5‐fluorouracil (GlcFU) and its 3′‐CH₂OH glucopyranose derivative. Both ligands were found to be low micromolar inhibitors with K_i values of 7.9 and 27.1 μm , respectively. X‐ray crystallography revealed that the 3′‐CH₂OH glucopyranose substituent is indeed involved in additional molecular interactions with the PLP γ‐phosphate compared with GlcFU. However, it is 3.4 times less potent. To elucidate this discovery, docking followed by postdocking Quantum Mechanics/Molecular Mechanics – Poisson–Boltzmann Surface Area (QM/MM‐PBSA) binding affinity calculations were performed. While the docking predictions failed to reflect the kinetic results, the QM/MM‐PBSA revealed that the desolvation energy cost for binding of the 3′‐CH₂OH‐substituted glucopyranose derivative out‐weigh the enthalpy gains from the extra contacts formed. The benefits of performing postdocking calculations employing a more accurate solvation model and the QM/MM‐PBSA methodology in lead optimization are therefore highlighted, specifically when the role of a highly polar/charged binding interface is significant.     

Influence of infused cell dose and HLA match on engraftment after double-unit cord blood allografts

The influence of cell dose and human leukocyte antigen (HLA) match on double-unit cord blood (CB) engraftment is not established. Therefore, we analyzed the impact of cell dose and high-resolution HLA match on neutrophil engraftment in 84 double-unit CB transplant recipients. The 94% sustained engraftment rate was accounted for by 1 unit in nearly all patients. Higher CD3(+) cell doses (P = .04) and percentage of CD34(+) cell viability (P = .008) were associated with unit dominance. After myeloablative conditioning, higher dominant unit total nucleated cell (TNC), CD34(+) cell, and colony-forming unit doses were associated with higher sustained engraftment and faster neutrophil recovery (P = .07, P = .0008, and P < .0001, respectively). Total infused TNC (P = .0007) and CD3(+) cell doses (P = .001) also significantly influenced engraftment. At high-resolution extensive donor-recipient HLA disparity was frequent, but had no influence on engraftment (P = .66), or unit dominance (P = .13). Although the unit-unit HLA match also did not affect sustained engraftment (P = 1.0), recipients of units closely (7-10 to 10-10) HLA-matched to each other were more likely to demonstrate initial engraftment of both units (P < .0001). Our findings have important implications for unit selection and provide further insight into double-unit biology.