Prediction of maximum scaled feed intake in broiler chickens based on physical properties of bulky feeds.

1. A trial was conducted to investigate the capacity of broiler chickens to consume bulky feeds during three stages of growth. These phases were from 1 to 15 d, 16 to 30 d and from 31 to 45 d. 2. A basal feed was serially diluted (0%, 2.5%, 5.0%, 10% or 15%) with one of five diluents (cellulose fibre, sawdust, rice husk, sand or vermiculite) to produce 25 feeds which were supplied on an ad libitum basis to the birds in each phase. Cobb 500® strain chicks were used, and, within each phase, each feed was given to nine individually-caged birds, 225 in total, distributed in a completely randomised design. 3. Intake increased initially, and then declined, as the proportion of each diluent increased. The consumption of feeds that limited intake were directly proportional to metabolic body weight and so a scaled feed intake, expressed as g/BW0.67 per day, was calculated. There were large effects of feed type on intake, in the short term, with consumption of a bulky feed leading to higher intakes. 4. It was concluded the Water Holding Capacity (WHC) content of the feeds could be appropriate measurement of 'bulk' responsible for limiting intake and could be used to predict maximum feed intake capacities of broiler chickens fed bulky diets.

Prolonged clinical remissions in patients with relapsed or refractory follicular lymphoma treated with autologous stem cell transplantation incorporating rituximab.

Three sequential phase II trials were conducted with different immunotherapy approaches to enhance the outcome of autologous transplant (high-dose therapy and autologous stem cell transplantation (HDT/ASCT)) for recurrent follicular lymphoma. Seventy-three patients were enrolled from 1996 to 2009. Patients received HDT/ASCT combined with (1) interferon-α 3 MU/m(2) subcutaneously (SC) three times per week (TIW) for 2 years post-ASCT, (2) rituximab (R) 375 mg/m(2) for in vivo purging 3-5 days pre-stem cell collection and 2 × 4 weekly R at 2 and 6 months post-ASCT, respectively, or (3) three infusions of R pre-stem cell collection followed by 6× R weekly and interferon-α 3 MU/m(2) SC TIW. Although not statistically significant, progression-free survival (PFS) for patients who received rituximab was 56.4 and 49.1% at 5 and 10 years compared to 36 and 21% in those who did not receive rituximab. Molecular relapse post-HDT/ASCT was the strongest predictor of PFS in a multivariate analysis. Molecular relapse was coincident with or preceded clinical relapses in 84% of patients who relapsed­median of 12 months (range 0-129 months). Adverse events included secondary malignancy, transformation to diffuse large B cell lymphoma, prolonged mostly asymptomatic hypogammaglobulinemia, and pulmonary fibrosis. The long-term toxicity profile must be considered when selecting patients for this treatment.

Prolonged in vitro hemolysis of EDTA-anticoagulated blood after a delayed hemolytic transfusion reaction.

BACKGROUND: A novel case where in vitro hemolysis was observed in plasma, but not in serum samples, obtained after the onset of a severe delayed hemolytic transfusion reaction is presented. CASE REPORT: A 54-year-old woman received 2 units of blood during an orthopedic procedure. She had received transfusion 30 years earlier, and testing before transfusion revealed no alloantibodies. The patient returned 12 days after the transfusion with a Hb level of 54 g per L due to a severe delayed hemolytic reaction caused by anti-K. The plasma and serum samples were grossly hemolysed on Day 12. On Day 14, the serum samples showed no evidence of hemolysis; however, the EDTA sample remained grossly hemolysed. This discrepancy was not identified until Day 19. Due to concerns of ongoing apparent severe hemolysis, the patient was unnecessarily treated with IVIG and corticosteroids. The in vitro hemolysis was still present at 75 days, despite complete normalization of her Hb, bilirubin, and LDH levels. The phenomenon had resolved by 125 days. CONCLUSION: This in vitro artifact has not been previously reported and the mechanism remains unclear. Both plasma and serum samples should be observed for hemolysis when evaluating a patient with a severe delayed hemolytic transfusion reaction.

Reporting of near-miss events for transfusion medicine: improving transfusion safety.

BACKGROUND: Half of the reported serious adverse events from transfusion are a consequence of medical error. A no-fault medical-event reporting system for transfusion medicine (MERS-TM) was developed to capture and analyze both near-miss and actual transfusion-related errors. STUDY DESIGN AND METHODS: A prospective audit of transfusion-related errors was performed to determine the ability of MERS-TM to identify the frequency and patterns of errors. RESULTS: Events and near-miss events (total, 819) were recorded for a period of 19 months (median, 51/month). No serious adverse patient outcome occurred, despite these events, with the transfusion of 17,465 units of RBCs. Sixty-one events (7.4%) were potentially life-threatening or could have led to permanent injury (severity Level 1). Of most concern were 3 samples collected from the wrong patient, 13 mislabeled samples, and 22 requests for blood for the wrong patient. Near-miss events were five times more frequent than actual transfusion errors, and 68 percent of errors were detected before blood was issued. Sixty-one percent of events originated from patient areas, 35 percent from the blood bank, and 4 percent from the blood supplier or other hospitals. Repeat collection was required for 1 of every 94 samples, and 1 in 346 requests for blood components was incorrect. Education of nurses and alterations to blood bank forms were not by themselves effective in reducing severe errors. An artifactual 50-percent reduction in the number of errors reported was noted during a 6-month period when two chief members of the event-reporting team were on temporary leave. CONCLUSION: The MERS-TM allowed the recognition and analysis of errors, determination of patterns of errors, and monitoring for changes in frequency after corrective action was implemented. Although no permanent injury resulted from the 819 events, innovative mechanisms must be designed to prevent these errors, instead of relying on faulty informal checks to capture errors after they occur.

Flow cytometry in the diagnosis of peritoneal carcinomatosis.

OBJECTIVES: Peritoneal carcinomatosis is the second major cause of ascites. Because of its frequency and poor prognosis, it is important to establish an accurate diagnosis. The aim of this study was to analyze the use of a DNA index, detemined by flow cytometry in the differential diagnosis of ascites, and to compare it to the cytopathological examination. METHODS: A prospective analysis was carried out on 67 patients (39 female, 28 male; mean age, 53+/-14 yr [range, 5-82]) with ascites of various etiologies. Peritoneal carcinomatosis was detected in 21 patients, whereas in 46 the ascites was of noncarcinomatosis origin. RESULTS: The sensitivity of the cytopathological examination for the diagnosis of peritoneal carcinomatosis was 42.9%, and the specificity was 100%. The mean DNA index determined by flow cytometry was similar for peritoneal carcinomatosis and noncarcinomatosis patients, being 1.28 versus 1.01, respectively, in the preparations without control lymphocytes and 1.28 versus 1.04, respectively, when control lymphocytes were added. The sensitivity of DNA index cytometry was 57.1% and specificity, 93.5%. The combined use of the DNA index and cytopathological examination did not show an advantage over the use of any of the tests individually, although the DNA index was able to detect half of the cases of peritoneal carcinomatosis in which cytopathological examination was negative. Although the sensitivity was higher when the parameters were associated, the DNA index did not offer a statistically significant advantage over the use of cytopathological examination alone, which in turn had higher specificity. CONCLUSION: The DNA index presented lower sensitivity for the diagnosis of peritoneal carcinomatosis when used alone, showing no advantage over conventional cytopathological examination. However, the DNA index was able to detect 50.0% of peritoneal carcinomatosis cases whose conventional cytopathological examinations were negative, and could be valuable in these situations.

Induction of tolerance to immunogenic tumor antigens associated with lymphomagenesis in HOX11 transgenic mice.

Transgenic mice expressing human HOX11 in B lymphocytes die prematurely from lymphomas that initiate in the spleen and frequently disseminate to distant sites. Preneoplastic hematopoiesis in these mice is unperturbed. We now report that expression of the HOX11 transgene does not affect the ability of dendritic cells (DCs) to process and present foreign peptides and activate antigen-specific T cell responses. We also show that nontransgenic DCs presenting peptides derived from the human HOX11 protein are highly efficient stimulators of autologous T cells, whereas transgenic T cells are nonresponsive to peptides derived from the HOX11 transgene and the murine Meis1 protein. HOX11 transgenic mice thus show normal development of tolerance to immunogenic antigens expressed throughout B cell maturation. DCs pulsed with cell lysates prepared from lymphomas, obtained from HOX11 transgenic mice with terminal lymphoma, activate T cells from nontransgenic and premalignant transgenic mice, whereas T cells isolated from lymphomatous transgenic mice are nonresponsive to autologous tumor cell antigens. These data indicate that HOX11 lymphoma cells express tumor-rejection antigens that are recognized as foreign in healthy transgenic mice and that lymphomagenesis is associated with the induction of anergy to tumor antigen-specific T cells. These findings are highly relevant for the development of immunotherapeutic protocols for the treatment of lymphoma.

Screening for azathioprine toxicity: a pharmacoeconomic analysis based on a target case.

The risk of azathioprine-induced myelosuppression can be predicted by detecting patients with intermediate or low thiopurine methyltransferase (TPMT) activity. Population studies have shown that 89% of whites have high TPMT activity, 11% have intermediate TPMT activity, and 0.3% have low TPMT activity. Three specific mutations in the TPMT gene that cause decreased TPMT activity have recently been identified. Patients homozygous for the TPMT mutations have low TPMT activity, and patients heterozygous for TPMT mutations have intermediate TPMT activity. This has led to the development of a technique for TPMT genotype analysis that will identify patients at risk of azathioprine-induced myelosuppression. We report a case of a patient with bullous pemphigoid who experienced azathioprine-induced myelosuppression and who was later found to be homozygous for TPMT mutant alleles. Using the cost of treatment required for this patient and the estimated population prevalence of TPMT mutations, we examined the cost impact of screening for TPMT mutations in all patients being considered for azathioprine therapy. We calculated that screening is cost-neutral assuming patients homozygous for TPMT mutations experience myelosuppression, and that it is cost-beneficial assuming patients heterozygous for TPMT mutations also experience myelosuppression while receiving azathioprine. Screening patients for TPMT mutations will reduce the risk of azathioprine-induced myelosuppression, and our study suggests that it may also be a cost-attractive strategy.

A gel technology system to determine postpartum RhIG dosage.

Failures of Rh immune globulin (RhIG) prophylaxis occur when the dose is too small. We report a test using a gel technology (GT) method to replace the Kleihauer-Betke (K-B) test to assess fetomaternal hemorrhage (FMH) and assist in determining the minimum necessary dose of RhIG. Cord blood (O, D+) was mixed with adult blood (O D-) to mimic an FMH of 10 mL, 20 mL, 28 mL, and 40 mL. Test samples were incubated with anti-D at known concentrations and centrifuged. The supernatant was titrated against D+ and D- red cells using GT and an interpretation of the required RhIG dose was made. Results were compared with the K-B test. Results were easily discernible and interpretations leading to determination of recommended RhIG dosage were reproducible. Correlation to standard K-B testing was confirmed. Elapsed time for result availability by GT testing was 60 minutes, with a direct technical time requirement of 30 minutes. The GT system is easier, objective, and quantitative, and compares well to the standard K-B test. A single procedure will allow assessment of the extent of FMH in the great majority of cases. This technique works well in determining the appropriate dose of anti-D required to treat D- patients with D+ newborns. There are potential cost savings in decreased use of RhIG, less direct technical time required, and more rapid availability of results.

A model for spontaneous B-lineage lymphomas in IgHmu-HOX11 transgenic mice.

HOX11, a divergent homeodomain-containing transcription factor, was isolated from the breakpoint of the nonrandom t(10;14)(q24;q11) chromosome translocation found in human T cell acute lymphoblastic leukemias. The translocation places the HOX11 coding sequence under the transcriptional control of TCR alpha/delta regulatory elements, resulting in ectopic expression of a normal HOX11 protein in thymocytes. To investigate the oncogenic potential of HOX11, we targeted its expression in lymphocytes of transgenic mice by placing the human cellular DNA under the transcriptional control of Ig heavy chain or LCK regulatory sequences. Only IgHmu-HOX11 mice expressing low levels of HOX11 were viable. During their second year of life, all HOX11 transgenic mice became terminally ill with more than 75% developing large cell lymphomas in the spleen, which frequently disseminated to thymus, lymph nodes, and other nonhematopoietic tissues. Lymphoma cells were predominantly clonal IgM+IgD+ mature B cells. Repopulation of severe combined immunodeficient mice with cells from hyperplastic spleens indicated that the HOX11 tumor phenotype was transplantable. Before tumor development, expression of the transgene did not result in perturbations in lymphopoiesis; however, lymphoid hyperplasia involving the splenic marginal zones was present in 20% of spleens. Our studies provide direct evidence that expression of HOX11 in lymphocytes leads to malignant transformation. These mice are a useful model system to study mechanisms involved in transformation from B-lineage hyperplasia to malignant lymphoma and for testing novel approaches to therapy. They represent a novel animal model for non-Hodgkin's lymphoma of peripheral mature B cell origin.

Verrucous cyst.

A verrucous cyst is an unusual, histopathologically distinctive, epidermoid cyst characterised by verrucous changes in its wall. We report two cases of verrucous cysts in different patients, one on the back and the other on the cheek. Clinically, the lesions were thought to represent an epidermoid cyst and a basal cell carcinoma, respectively. Histologically, we found in both cases an intradermal epidermoid cyst lined by a stratified squamous epithelium with focal cytopathogenic viral changes, consisting of papillomatosis, ortho- and parakeratosis, and hypergranulosis. One case also showed, within the squamous areas of the hyperplastic epithelium, occasional squamous eddies. These histopathological features support the diagnosis of verrucous cyst, which may represent a manifestation of human papillomavirus infection. This virus may induce cyst formation or just infect a pre-existing one.

Changes in activated protein C resistance during normal pregnancy.

OBJECTIVE: The objective of this study was to determine the changes in activated protein C resistance that occur during normal pregnancy. STUDY DESIGN: In this cross-sectional study activated protein C was measured in 128 women with normal pregnancies in the first, second, and third trimesters and in nonpregnant control subjects with 24 to 39 women in each group. In addition, factor V, factor VIII, free protein S, and functional protein C were measured and correlated with activated protein C levels. Polymerase chain reaction for factor V Leiden mutation was performed. RESULTS: There was a significant fall in the activity of activated protein C in the second and third trimesters of pregnancy (p < 0.05). This was related to increased factor VIII and decreased free protein S levels (p = 0.002, R2 = 0.20). The prevalence of the factor V Leiden mutation was 7.3%. CONCLUSION: Resistance to activated protein C is increased in the second and third trimesters of pregnancy. This is related to the alterations in other coagulation proteins, a phenomenon normally occurring during pregnancy.

Loss of functional cell surface transforming growth factor beta (TGF-beta) type 1 receptor correlates with insensitivity to TGF-beta in chronic lymphocytic leukemia.

Chronic lymphocytic leukemia (CLL) is the most common form of adult leukemia in Western countries, and there is significant variability in survival within CLL clinical stages. Earlier studies showed that CLL cells produce and are usually growth inhibited by transforming growth factor beta type 1 (TGF-beta1), suggesting a mechanism for the clinically indolent course of most CLL. Here we studied the mechanism by which CLL cells from about one-third of the patients are insensitive to TGF-beta1. Of the 13 patients studied, CLL cells isolated from the peripheral blood of 8 patients were sensitive to growth inhibition by TGF-beta1, as determined by incorporation of tritiated thymidine, whereas those from 5 patients were completely resistant to TGF-beta1. As judged by binding of radiolabeled TGF-beta1 followed by cross-linking and immunoprecipitation with anti-receptor antisera, CLL cells sensitive to TGF-beta1 exhibited normal cell surface expression of both types 1 and 2 TGF-beta receptors. In contrast, all CLL cells resistant to TGF-beta1 exhibited no detectable surface type I receptors able to bind TGF-beta1, but normal expression of type II receptors. Both TGF-beta1-sensitive and TGF-beta1-resistant CLL cells contained normal amounts of both type 1 and type 2 receptor mRNAs. Specific loss of type 1 receptor expression represents a new mechanism by which cells acquire resistance to TGF-beta1-mediated growth inhibition in the development and progression of human lymphoproliferative malignancies.

Transforming function of the HOX11/TCL3 homeobox gene.

The HOX11/TCL3 homeobox gene was identified at the breakpoint region in pediatric T-cell acute lymphoblastic leukemia harboring 10q24 chromosomal translocations. We previously reported that primary murine bone marrow cells transduced ex vivo with a recombinant HOX11-containing retrovirus, MSCV-HOX11, gave rise to cell lines at high frequency having characteristics of early myeloid cells. Cell lines were also established from the bone marrow and spleen of transplant recipients sacrificed 5 months after engraftment with MSCV-HOX11-transduced bone marrow cells. These latter lines, which exhibited a more differentiated myelomonocytic phenotype, harbored proviruses encoding a smaller HOX11 protein. None of the mice that received HOX11-expressing bone marrow cells or myeloid cell lines developed leukemia during 6-month observation periods. Here, we report that two bone marrow transplant recipients eventually developed T-cell acute lymphoblastic leukemia-like malignancies at 7 and 12 months posttransplant, indicating that progression to a fully malignant state required additional mutations. One tumor synthesized full-length HOX11 whereas the other expressed the smaller version of the protein. The smaller HOX11 protein suffered a carboxyl-terminal truncation. We subsequently constructed MSCV-based retroviral vectors expressing deleted forms of HOX11 and identified an amino-terminal region that was dispensible for generation of myeloid cell lines having a similar phenotype as those induced by full-length HOX11. We thus conclude that regions near the amino and carboxyl termini of HOX11 are not essential for transforming function, nor do they appear to determine the lineage or stage of differentiation of the target cell for transformation.

Acute gouty synovitis associated with "urate milk".

OBJECTIVE: To analyze the clinical features of acute gouty synovitis associated with thick, milky white, "chalky," urate laden synovial effusions, and to investigate the effects on synovial white blood cell (WBC) counts when leukocyte-rich rheumatoid effusions are incubated with a urate packed milky synovial fluid. METHODS: Five patients (all men, mean age 70.8 years) with acute gouty synovitis (acute arthritis in 3, acute bursitis in 2) associated with "urate milk" were studied between 1993 and 1996. RESULTS: Synovial effusions were thick, "chalky," and appeared "milky" white. The fluids were packed with monosodium urate (MSU) crystals, which sedimented upon standing, leaving a clear supernatant containing a few MSU crystals. The presence of massive amounts of MSU crystals and crystal clumps interfered with accurate determination of synovial WBC counts. Four fluids showed "a few leukocytes," and one a WBC count of 6750/mm3 with 91% neutrophils and several intraleukocytic crystals. Four patients had subcutaneous tophi. Of the risk factors associated with development of gout, the most frequent was ethanol abuse, in 4 and possibly all 5 patients. Incubation of leukocyte-rich rheumatoid synovial effusions with urate laden knee fluid from Patient 5 produced a greater reduction in synovial WBC counts compared to controls. CONCLUSION: Milky white synovial effusions containing massive quantities of urate crystals (referred to as "urate milk") may rarely occur in the setting of acute gouty arthritis or bursitis. Ethanol abuse appears to be a risk factor associated with the development of hyperuricemia and gout in these patients.

[Dermatitis artefacta]. / Dermatite artefacta.

We report a case of a 29-year-old man presenting skin ulcerations on both sides of the mandible. The diagnosis of dermatitis artefacta was based on the morphology and evolution of the lesions, on the patient's borderline personality, on the objects found in his possession, and a later admission of having an involvement in the aggravation of the lesions.

Immunosuppressive toxicity of CAMPATH1H monoclonal antibody in the treatment of patients with recurrent low grade lymphoma.

We have studied, as part of a group of international multicenter phase II clinical trials, the toxicity and effectiveness of CAMPATH1H administered intravenously three times a week in an outpatient setting to patients with recurrent or progressive low grade lymphoma. We report here on the toxicity and therapeutic results of the first seven patients treated before the study was closed prematurely because of unacceptable toxicity. Classical complete or partial responses of treatment were seen in three of seven patients. One complete response lasted 8.5 months and the other complete response is ongoing at 1 year. Responses occurred in nodal sites as well as in skin and peripheral blood. The first three or four antibody infusions in each patient was associated with grade 1 or 2 side-effects including rigor, fever, facial flushing, nausea, vomiting, hives, wheezes, hypotension, and/or diarrhea but these subsequently decreased or disappeared. The most significant toxicity was profound lymphopenia and associated infection, usually viral. Six of seven patients had culture or serologically documented infections and four patients had two or more such episodes. All infections responded to temporary discontinuation of antibody therapy and appropriate antiviral or antibiotic agents. We conclude that CAMPATH1H monoclonal antibody has therapeutic activity against low grade non-Hodgkin's lymphoma but that this activity is limited by marked lymphopenia and an unacceptably high frequency of serious infection at the dose and schedule used in this trial.

SBH-1, a novel Reed-Sternberg-like cell line capable of inducing tumors in SCID mice: immunophenotypic, cytogenetic, and cytokine expression profiles.

A new cell line, SBH-1, with the morphologic, immunophenotypic, and karyotypic features consistent with those of Reed-Sternberg (RS) and Hodgkin (H) cells, has been established from the pleural effusion of a patient. The cytologic appearance of SBH-1 cells is characteristic of multinucleate RS and mononuclear H cells, all containing inclusion-like nucleoli. The SBH-1 cells express CD30, CD15, CD25, CD71, CD45, CD20, CD22, and bcl-2 protein and are negative for epithelial membrane antigen. Cytogenetic analysis showed multiple clonal abnormalities with breakpoints at 14q32, 6q21, and 11q23. The Ig heavy chain genes and both Ig light chain genes were rearranged in SBH-1 cells, whereas the bcl-2 gene was in germline configuration. Messages for the cytokines interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha, and transforming growth factor-beta and the cytokine receptors IL-2R, IL-4R, IL-6R, and IL-7R were detected by reverse transcription-polymerase chain reaction analysis. Xenotransplantation of SBH-1 cells into severe combined immunodeficient (SCID) mice led to local and disseminated tumor growth. The cytologic, histologic, and immunohistochemical features of SBH-1 cells in SCID mouse tumors were typical of RS and H cells. The SBH-1 cell line will be useful in the study of RS and H cell biology, inasmuch as it represents a cell line obtained from a previously untreated patient.