Tissue inhibitor of metalloproteinases (TIMP)-1, -2 and -3 in human endometrium during the menstrual cycle

The extensive remodelling of the human endometrium throughout the menstrual cycle is accompanied by changes in production of matrix metalloproteinases, the activity of which can be inhibited by specific tissue inhibitors or by tissue inhibitors of metalloproteinases (TIMP)s with a 1:1 stoichiometry. This study immunolocalized TIMP-1, TIMP-2 and TIMP-3 in dated normal human endometrium across the menstrual cycle and examined cultured endometrial cells for their production. All three TIMPs were present in the major cellular compartments, luminal epithelium, glands, stroma, endothelial cells and vascular smooth muscle cells with the most intense immunoreactivity in the luminal epithelium. TIMP-1 and -3 were lower in the mid-to-late proliferative phase with a nadir of TIMP-3 particularly in the late proliferative phase. Decidualized stromal cells stained strongly positive for TIMP-1, -2 and -3. Cells of haematopoietic origin never stained. Monensin treatment of tissue resulted in accumulation of TIMPs in all cellular compartments but particularly of TIMP-1 in epithelium. Cultured endometrial stromal cells released more TIMP-1 than TIMP-2 or TIMP-3 into culture medium and all were increased following decidualization in vitro. Epithelial cells in culture produced less TIMPs than stromal cells, and only a few epithelial cells in each culture were immunopositive for TIMP-1. The ubiquitous distribution of TIMPs implicates them in maintenance of endometrial integrity, with changes in the matrix metalloproteinases without concomitant changes in TIMPs determining endometrial matrix degradation.      

The detection of platelet alloantibodies by flow cytometry

A flow cytometric procedure was investigated for its ability to detect antibodies directed against blood group A, HLA, and PlA1 (HPA-1a) antigens. When type O sera were tested against platelets from blood group A donors, only 9 of 14 positive reactions were observed. Furthermore, the expression of blood group A varied more than 100-fold on platelets derived from individual donors. When anti-HLA-A2 and -B7 were evaluated, 11 of 11 individuals with HLA-A2 and -B7 antigens reacted. In contrast, when platelets from donors whose HLA antigens included HLA-B8 or -B12 were tested with anti-HLA-B8 or -HLA-B12, respectively, positive reactions were observed in only 3 of 7 instances, despite the fact that the lymphocytes reacted strongly. Platelets from 10 HLA-A2-positive donors, which had been stored for up to 20 months at -70 degrees C, were studied. In all cases, frozen-stored platelets reacted well with an anti-HLA-A2. Limited testing with an anti-PlA1 (anti-HPA-1a) showed equal reactivity with fresh and frozen platelets. Finally, the method was compared to a visual immunofluorescence assay using sera from patients who were refractory to platelet transfusions. The results agreed in 30 of 37 comparisons, and most discrepancies were resolved in favor of flow cytometry. It is concluded that flow cytometry is useful for detecting platelet alloantibodies and possibly for prospective platelet crossmatching, as HLA- and platelet-specific antibodies can be identified by using platelets stored frozen for several months.     

Pro/con ethics debate: is nonheart-beating organ donation ethically acceptable?

This pro/con debate explores the ethical issues surrounding nonheart-beating organ donation (NHBD), a source of considerable controversy. It is estimated that NHBD can increase the number of organs available for transplant by 25% at a time of great need. However, should NHBD be ethically acceptable? In support of NHBD, it may be acceptable practice if there is a separation of the rationale to withdraw life support/to withhold cardiopulmonary resuscitation from the decision to recover organs, if no conflicts of interest exist, if a waiting time precluding spontaneous return of circulation is included, and if NHBD conforms to a standardized protocol. Against NHBD, there are questions regarding the ambiguity and cultural perspectives of death, regarding whether a separation of rationale between withdrawal and donation is sufficient to preclude conflicts of interest, and regarding whether variable protocols arise that subordinate the patient to the goal of donation. Such concerns suggest NHBD may damage the trust in patient-physician relationships and may adversely affect organ donation rates.     

Viability and functional integrity of washed platelets

The viability and functional integrity of saline- and ACD-saline-washed platelets were compared with those of unwashed platelets. After template bleeding time (TBT) was measured, 15 healthy volunteers underwent plateletpheresis and ingested 600 mg of aspirin. Autologous 111In-labeled platelets were transfused: unwashed (n = 5), washed with 0.9 percent saline solution (SS) (n = 5), and washed with a buffered 12.6 percent solution of ACD-A in 0.9 percent saline solution (n = 5). After transfusion, we measured TBT at 1, 4, and 24 hours; platelet survival at 10 minutes and 1, 4, and 24 hours and daily for 6 days; and the percentage of uptake in liver and spleen by quantitative whole-body radionuclide scintigraphy at 24 and 190 hours. We found that saline washing affected platelet recovery, 23.47 +/- 12 percent (p less than 0.001) as compared to 52.43 +/- 17 percent (p less than 0.002) for ACD-saline and 73.17 +/- 8 percent for control; that saline washing resulted in a greater liver uptake than control and ACD-saline-washed platelets (31.9 +/- 8% [p less than 0.001] vs 17.7 +/- 4.1 and 19.3 +/- 2.1% [p greater than 0.1], respectively); that, unlike control and ACD-saline-washed platelets, saline-washed platelets did not shorten bleeding time; and that neither type of washing affected survival. Although ACD-saline washing affects recovery, it also results in intact function, normal survival, higher recovery than SS platelets, and no significant liver uptake.     

Composition of peripheral blood progenitor cell components collected from healthy donors

BACKGROUND: Peripheral blood progenitor cell (PBPC) components are being collected from healthy donors for allogeneic transplantation, but the quantity, quality, composition, and variability of PBPCs collected from healthy people given granulocyte-colony-stimulating factor (G-CSF) have not been evaluated. STUDY DESIGN AND METHODS: PBPC components were collected from 150 healthy people who were given G-CSF (5, 7.5, or 10 microg/kg/day) for 5 days. The components were evaluated for white cell (WBC), mononuclear cell, CD34+ cell, neutrophil, platelet, and red cell (RBC) composition. RESULTS: The quantities collected were: WBCs, 35.0 +/? 16.4 × 10(9) (range, 11.9–163.3 × 10(9)); mononuclear cells, 33.3 +/? 14.4 × 10(9) (range, 11.9–139.6 × 10(9)); CD34+ cells, 412 +/? 287 × 10(6) (range, 70–1658 × 10(6)); neutrophils, 1.71 +/? 3.59 × 10(9) (range, 0–27.6 × 10(9)); RBCs, 7.2 +/? 4.0 mL (range, 0–22.1 mL); and platelets, 480 +/? 110 × 10(9) (range, 250–920 × 10(9)). PBPC components collected from people given G-CSF at 7.5 or 10 microg per kg per day contained significantly more CD34+ cells (respectively, 428 +/? 300 × 10(6); range, 70–1658 × 10(6) and 452 +/? 294 × 10(6); range, 78- 1380 × 10(6)) than those from people given G-CSF at 5 microg per kg per day (276 +/? 186 × 10(6); range, 91–767 × 10(6)) (p = 0.007 and p = 0.002). When 10 microg per kg per day of G-CSF was given, 50 percent of the components contained enough CD34+ cells for transplantation to a 75- kg recipient (375 × 10(6) CD34+ cells), but 10.6 percent of the components contained less than 150 × 10(6) CD34+ cells and thus would provide a transplantable dose only for a 30-kg patient. CONCLUSION: One PBPC component collected from a healthy donor given 7.5 or 10 microg per kg per day of G-CSF should contain 70 to 1660 × 10(6) CD34+ cells, with 0 to 22 mL of RBCs. Because of the great variability in the number of CD34+ cells collected, the quantity of CD34+ cells in each component should be measured after each procedure to ensure that sufficient quantities of cells are present for a successful transplant.     

Cell elastimetry in the detection of antineutrophil antibodies

Cell elastimetry has been applied to the measurement of antineutrophil antibodies. This technique measures, under direct visualization, the negative pressure required of aspirate PMNs into small-pored pipettes. Two groups of studies were carried out: (A) In the first group of studies, normal PMNs were incubated with 1 of 8 known antineutrophil serums. Each serum significantly decreased membrane deformability-- i.e., cells became more rigid. The study was conducted in an entirely blind fashion. Randomly coded serums from patients and controls were studied for deformability by observers unaware of the code. (B) In the second group of studies, sera containing immune complexes were incubated with normal PMNs. No significant effects were noted upon deformability. As a single cell assay that partially reflects membrane rigidity, elastimetry may, therefore, have potential in the further characterization of mechanisms by which such antineutrophil antibodies compromise neutrophil functions.     

Value of radiopaque markers in identifying partial small bowel obstruction

Confirming partial small bowel obstruction is often a diagnostic challenge. In this case report, 4-mm solid radiopaque markers were used in 4 patients to show partial small bowel obstruction. Results of enteroclysis were normal in 2 of the 4 patients, and the markers were used to challenge suspected partial obstruction. The markers coalesced in the region of the partial obstruction, which was confirmed at surgery. Enteroclysis is the examination of choice in the diagnosis of partial small bowel obstruction. However, examinations with false- negative results can occur, particularly with adhesive and/or intermittent obstructions. The use of radiopaque markers in these cases proved an effective and useful method of establishing the diagnosis of partial small bowel obstruction, particularly in the 2 cases in which enteroclysis results were normal. Prospective studies are needed to establish the feasibility of this novel technique. (Gastroenterology 1996 Jun;110(6):1958-63)     

Febrile reactions after platelet transfusion: the effect of single versus multiple donors

Febrile reactions to platelet transfusions are a common problem. The platelet transfusion records from a 30-month period were analyzed to determine 1) when reactions occur in a transfusion sequence; 2) how frequently they recur; and 3) whether the choice of multiple-donor (pooled concentrates) or single-donor components (unmatched apheresis and HLA-compatible apheresis platelets) affected the reaction rate. Overall, 18.7 percent of all patients receiving platelets experienced reactions. A subset of 85 patients, who began platelet support with unmodified components during the study interval, were analyzed in detail. This group received 1204 unmodified transfusions (mean, 14.2/patient), which were associated with 171 reactions (per-transfusion reaction rate, 14.2%). Despite a higher mean white cell content, the transfusion of 438 unmatched single-donor platelets (10.84 x 10(8) white cells, 8.4% reaction rate) resulted in reactions significantly less often than did that of 583 pooled concentrates (8.53 x 10(8) white cells, 21.4% reaction rate) (p less than 0.001). The rate of reaction to HLA-compatible platelets (9/183 transfusions, 4.9%) was not significantly different from that to unmatched single-donor platelets. The use of platelet components from one donor, as opposed to multiple donors, may provide an effective means of reducing the incidence of febrile reactions.