Umbilical cord blood transplant from unrelated HLA-mismatched donors in children with high risk leukemia.
In the last 3 years, 14 children with high-risk leukemia (11 ALL, 2 AML and 1 CML) underwent cord blood transplantation from unrelated HLA-mismatched donors at a median of 99 days from the start of search. Eight patients were transplanted in second CR, one in accelerated phase, three at relapse and two patients in first CR. Conditioning regimen (fractionated TBI, etoposide, CY and anti-lymphocyte serum) and prophylaxis of GVHD (CsA and 6-methylprednisolone) were identical for all patients. Neutrophils >0.5x10(9)/l were reached at a median of 33 days from transplant, but in four cases we observed an autologous hematopoietic reconstitution (three spontaneous, one after autologous BM rescue). Acute and chronic GVHD were observed in 10/14 and 3/8 evaluable cases, respectively. Three patients died of transplant-related toxicity and three patients relapsed. The probabilities of event-free, disease-free and overall survival were 50, 53 and 64%, respectively. Cord blood transplant from HLA-mismatched unrelated donor is a valid option for the treatment of children with high-risk leukemia. With our eligibility criteria, conditioning regimen and prophylaxis of graft-versus-host disease, the main obstacles to successful transplant were represented by graft failure and fatal acute GVHD. (+info)
Supplying commercial biomedical companies from a human tissue bank in an NHS hospital--a view from personal experience.
NHS histopathology laboratories are well placed to develop banks of surgically removed surplus human tissues to meet the increasing demands of commercial biomedical companies. The ultimate aim could be national network of non-profit making NHS tissue banks conforming to national minimum ethical, legal, and quality standards which could be monitored by local research ethics committees. The Nuffield report on bioethics provides ethical and legal guidance but we believe that the patient should be fully informed and the consent given explicit. Setting up a tissue bank requires enthusiasm, hard work, and determination as well as coordination between professionals in the NHS trust and in the commercial sector. The rewards are exiting new collaborations with commercial biomedical companies which could help secure our future. (+info)
Cancer risk estimates for family members of a population-based family registry for breast and ovarian cancer.
Population-based breast and ovarian cancer family registries can facilitate studies to evaluate genetic and environmental factors in the etiology of these malignancies. The purpose of this study is to describe what is, as far as we know, the first population-based breast and ovarian cancer family registry and to estimate breast and ovarian cancer risk in relatives of breast and ovarian cancer probands. Population-based consecutive incident cases of breast and ovarian cancer were invited to participate in the University of California, Irvine breast and ovarian family registry. In this study, we report data on 1567 breast cancer and 328 ovarian cancer probands. The operational components of this family registry include enrollment of probands, family history interviewing, confidentiality, pathology, verification and review, biospecimen bank, statistical/genetic analysis, and special studies on positional cloning of known genes. All of the components are tracked through the University of California, Irvine Genetic Research Information System. In non-Hispanic-white breast cancer probands, relative risk (RR) of breast cancer in mothers and sisters is significantly elevated [RR = 1.7 and 95% confidence interval (CI) = 1.4-2.0 and RR = 2.8 and 95% CI = 2.3-3.3, respectively]. In families of ovarian cancer probands, mothers are at increased risk of ovarian cancer (RR = 4.6; 95% CI, 2.1-8.7). RR of breast cancer in mothers of Hispanic breast cancer probands is significantly elevated (RR = 4.9; 95% CI, 2.6-8.5). No elevation of breast or ovarian cancer risk was observed among relatives of Asian probands. In general, there is a decrease in RR among mothers and sisters with increase in age of onset of probands. In second-degree relatives and first cousins, the breast cancer hazards ratios increase with increase in the number of affected first-degree relatives and decrease with increase in age at onset of the proband. (+info)
The mechanical properties of fresh and cryopreserved arterial homografts.
OBJECTIVES: To assess the effect of cryopreservation on the elasticity and compliance of arterial allografts. MATERIALS AND METHODS: Iliofemoral segments of arteries and veins harvested from multiorgan donors were divided into two groups: fresh-control, tested for 24 hours after harvesting, and cryopreserved in liquid nitrogen after pretreatment with 20% dimethylsulphoxide and stored for an average time of 22 days. Vessel wall elastic properties were evaluated from the stress-strain relationship in a specially designed test cell fixed to the Instron Universal Testing Machine. RESULTS: The elastic modulus of the artery control group (1.54+/-0.33 MPa, n=20) was not significantly different from the cryopreserved group (1.69+/-0.61 MPa, n=15). Similarly, values for unfrozen veins (3.11+/-0.65 MPa, n=47) were not significantly different from those of frozen samples (2.71+/-0.85 MPa, n=38). Control compliance (6. 86+/-1.79x10(-5)%/Pa, for arteries; 3.84+/-0.81x10(-5)%/Pa, for veins) was similar to that of the cryopreserved group (6.66+/-1. 80x10(-5)%/Pa, for arteries; 4.16+/-1.21x10(-5)%/Pa, for veins). CONCLUSIONS: Cryopreservation maintains the important elastic properties of arterial and venous allografts during average storage time of 22 days. (+info)
Cadaver donor screening for infectious agents in solid organ transplantation.
The transmission of infection by a cadaver donor organ can result not only in loss of the allograft but also in death of the immunosuppressed recipient. Despite the shortage of cadaver organ donors, every donor must be evaluated thoroughly for the potential transmission of infectious disease, because the consequences of the organ donor events can have a profound effect on the transplant outcome. This review summarizes current knowledge about serological screening of organ donors to determine the suitability of organs from cadaver donors for transplantation. (+info)
Polymerase chain reaction detection of Kaposi's sarcoma-associated herpesvirus-optimized protocols and their application to myeloma.
Since its discovery in 1994, KSHV (also called human herpesvirus-8 or HHV8) has been implicated in a variety of disorders. Although the association of KSHV with Kaposi's sarcoma (KS), primary effusion lymphoma (PEL), and multicentric Castleman's disease has been well established, its presence in some other diseases, such as multiple myeloma, remains controversial. Because most KSHV studies are based on polymerase chain reaction (PCR) analysis, the conflicting data may be attributable to variations in the methods, primer sets, and target sequences selected. To establish an efficient and reliable PCR approach for KSHV detection we designed eight sets of primers to six regions (ORFK1, ORFK2, ORFK9, ORK26, ORF72, and ORF74) of the KSHV genome using appropriate database and software. The detection sensitivity of these primers was carefully assessed and their reliability was strictly validated in a series of positive (15 KS and PEL samples) and negative (16 lymphoid tissues) controls. We found that primer sets to the ORFK9 region showed the highest sensitivity, whereas primer sets to ORFK1 and ORF74 showed the lowest sensitivity. Primer sets to ORFK9, ORF26 and ORF72 regions detected all of the positive cases, whereas other primer sets showed varying detection rates or nonspecific bands. All 16 negative controls were negative with all primer sets. However, six of 16 negative controls became positive when we used nested PCR targeting ORF26. Therefore, multiple target KSHV sequences increase the detection efficiency, while nested PCR protocols are likely to introduce false positivity. Using ORFK9, ORF26 and ORF72 primer sets, we screened bone marrow biopsies from 18 cases of multiple myeloma, and failed to detect any KSHV sequences. This finding supports the conclusion that KSHV is not associated with multiple myeloma. Indeed, our results further confirm that although KSHV is universally present in Kaposi's sarcoma and primary effusion lymphoma, it is not ubiquitious. (+info)
Preservation of RNA for functional genomic studies: a multidisciplinary tumor bank protocol.
Few human tumors are collected such that RNA is preserved for molecular analysis. Completion of the Human Genome Project will soon result in the identification of more than 100,000 new genes. Consequently, increasing attention is being diverted to identifying the function of these newly described genes. Here we describe a multidisciplinary tumor bank procurement protocol that preserves both the integrity of tissue for pathologic diagnosis, and the RNA for molecular analyses. Freshly excised normal skin was obtained from five patients undergoing wound reconstruction following Mohs micrographic surgery for cutaneous neoplasia. Tissues treated for 24 hours with RNAlater were compared histologically and immunohistochemically to tissues not treated with RNAlater. Immunohistochemical stains studied included: CD45, CEA, cytokeratin AE1/3, vimentin, S-100, and CD34 on formalin-fixed, paraffin embedded tissue and CD45 staining of frozen tissue. Slides were blinded and evaluated independently by three pathologists. The histologic and immunohistochemical parameters of tissue stored in RNAlater were indistinguishable from tissue processed in standard fashion with the exception of S-100 stain which failed to identify melanocytes or Langerhan's cells within the epidermis in any of the RNAlater-treated tissues. Interestingly, nerve trunks within the dermis stained appropriately for S-100. Multiple non-cutaneous autopsy tissues were treated with RNAlater, formalin, liquid nitrogen (LN2), and TRIzol Reagent. The pathologists were unable to distinguish between tissues treated with RNAlater, formalin, or frozen in LN2, but could easily distinguish tissues treated with TRIzol Reagent because of extensive cytolysis. RNA was isolated from a portion of the tissue treated with RNAlater and used for molecular studies including Northern blotting and microarray analysis. RNA was adequate for Northern blot analysis and mRNA purified from RNAlater-treated tissues consistently provided excellent templates for reverse transcription and subsequent microarray analysis. We conclude that tissues treated with RNAlater before routine processing are indistinguishable histologically and immunohistochemically from tissues processed in routine fashion and that the RNA isolated from these tissues is of high quality and can be used for molecular studies. Based on this study, we developed a multidisciplinary tumor bank procurement protocol in which fresh tissue from resection specimens are routinely stored in RNAlater at the time of preliminary dissection. Thus, precious human tissue can be utilized for functional genomic studies without compromising the tissue's diagnostic and prognostic qualities. (+info)
Exploring the host desmoplastic response to pancreatic carcinoma: gene expression of stromal and neoplastic cells at the site of primary invasion.
The dramatic opportunities presented by comprehensive gene profiling technologies are curbed by the problem of segregating these large amounts of gene expression data into meaningful categories for study. This is particularly evident in infiltrating carcinomas of the pancreas, in which global gene expression data primarily mirrors the prominent desmoplastic response to the infiltrating neoplasm. In an effort to better characterize the gene expression of invasive pancreatic cancers and their associated desmoplastic response, we performed in situ hybridization on pancreatic cancer tissues to characterize the expression of 12 genes identified by serial analysis of gene expression as highly expressed in invasive pancreatic cancer tissues but not in pancreatic cancer cell lines. In situ hybridization demonstrated that eight genes were expressed within the stromal and/or angioendothelial cells of the desmoplastic response to the invasive tumor, and four of these genes were specifically expressed by the stromal cells immediately adjacent to the invasive neoplastic epithelium, suggesting regional differences in gene expression within the host desmoplastic response. In contrast, four genes were specifically expressed by the invasive neoplastic epithelium, indicating important differences between in vivo and in vitro gene expression of human epithelial neoplasms. We have identified a highly organized structure of gene expression within the host stromal response to invasive pancreatic cancer that may reflect tumor-host communication and serve as a target for therapeutic intervention. (+info)