Gamma and X-ray irradiation do not affect the in vitro quality of refrigerated apheresis platelets in platelet additive solution (PAS-E).

BACKGROUND: Platelet refrigeration (cold storage) provides the advantages of an extended shelf life and reduces the risk of bacterial growth, compared to platelets stored at room temperature (RT). However, processing modifications, such as irradiation, may further improve the safety and/or alter the quality of cold-stored platelets. Platelet components are irradiated to prevent transfusion-associated graft versus host disease (TA-GvHD) in high-risk patients; and while irradiation has little effect on the quality of RT-stored platelet components, there is no data assessing the effect irradiation has following cold storage. STUDY DESIGN AND METHODS: Triple-dose apheresis platelets were collected in 40% plasma/60% PAS-E, using the TRIMA apheresis platform, and refrigerated (2-6°C) within 8 h of collection. On day 2, one of each component was gamma or X-ray irradiated or remained non-irradiated. Platelets were tested over 21 days. RESULTS: The platelet concentration decreased by approximately 20% in all groups during 21 days of storage (p > .05). Irradiation (gamma or X-ray) did not affect platelet metabolism, and the pH was maintained above the minimum specification (>6.4) for 21 days. The surface phenotype and the composition of the supernatant was similar in non-irradiated and irradiated platelets, regardless of the source of radiation. Functional responses (aggregation and clot formation) were not affected by irradiation. DISCUSSION: Gamma and X-ray irradiation do not affect the in vitro quality of platelet components stored in the cold for up to 21 days. This demonstrates the acceptability of irradiating cold-stored platelets, which has the potential to improve their safety for at-risk patient cohorts.

X-irradiation and gamma-irradiation inactivate lymphocytes in blood components.

BACKGROUND: Irradiation of selected blood components is standard practice for the prevention of transfusion-associated graft-versus-host disease (TA-GvHD). Currently, gamma-irradiation is the most widely used form of irradiation, but there is an increasing interest in X-irradiation, which is considered to be functionally equivalent and safer. However, there is a paucity of contemporary data regarding the ability of X-irradiation to inactivate lymphocytes in blood components. Therefore, the effect of gamma- and X-irradiation on lymphocyte viability and function in blood components was compared. STUDY DESIGN AND METHODS: Lymphocytes were isolated from venous blood by density gradient centrifugation, spiked into plasma/SSP+ to simulate a blood component, and either gamma- or X-irradiated. The phenotype of the isolated lymphocytes was confirmed. Lymphocyte viability was measured using a LIVE/DEAD assay, and function was assessed using mixed lymphocyte culture and CD69 expression post-phorbol-12 myristate 13-acetate (PMA) stimulation. RESULTS: Lymphocyte viability and CD69 expression following PMA stimulation were significantly reduced by both gamma-irradiation and X-irradiation in simulated blood components. Allorecognition and allostimulation were also significantly reduced by both gamma-irradiation and X-irradiation. CONCLUSION: Lymphocyte viability and function are reduced to a similar extent by gamma- and X-irradiation in simulated blood components. As such, X-irradiation is suitable for the irradiation of blood components and, in terms of lymphocyte inactivation, could be used instead of gamma-irradiation.

The in vitro quality of X-irradiated platelet components in PAS-E is equivalent to gamma-irradiated components.

BACKGROUND: Blood components are irradiated to inactivate lymphocytes in an effort to prevent transfusion-associated graft versus host disease. Although gamma irradiators are commonly used, they are subjected to rigorous health, safety, and compliance regulations, compared with X-irradiators which have the advantage of only emitting radiation while the machine is switched on. While the effects of gamma irradiation on platelet components are well known, there is little or no data comparing the effects of X- and gamma-irradiation on the quality of these components. Therefore, this study examined the in vitro quality of platelet components (pooled and apheresis) following X- or gamma-irradiation. STUDY DESIGN AND METHODS: Whole-blood-derived (pooled) and apheresis platelet components in platelet additive solution (n = 20 pairs for each type) were irradiated (X vs. gamma). In vitro platelet quality was tested prior to irradiation (day 1) and subsequently on days 2, 5, and 7. Non-irradiated components were tested on day 5 in parallel as reference controls. Metabolic parameters, surface expression of glycoproteins and activation markers (CD62P and annexin-V binding), and agonist-induced aggregation were measured. RESULTS: All components met Council of Europe specifications. There were no statistical differences in any in vitro quality measurements between X- and gamma-irradiated pooled or apheresis platelet components. CONCLUSION: X- and gamma-irradiation have similar effects on the in vitro quality of stored blood components, indicating that either technology represents a suitable option for irradiation of platelet components.

Hatching and Mortality of Meloidogyne enterolobii Under the Interference of Entomopathogenic Nematodes In vitro.

Plant parasitic nematodes have become one of the main problems in the tomato cultivation. Among these, Meloidogyne enterolobii presents great challenges to the farmer, since it is a polyphagous species and difficult to control. The entomopathogenic nematodes (EPNs) present as potential for biological control of this pathogen. The objective of the study was to evaluate the interference of EPNs S. brazilense, S. feltiae, S. rarum, H. amazonensis and H. bacteriophora on hatching and mortality of M. enterolobii. 500 eggs of this nematode and 1.000 infective juveniles of each EPN species were placed in a plastic pot totaling 25 mL of suspension and kept in an incubator at 25°C. The number of juveniles hatched in the suspension was counted every 2 days, until 10 days. After 10 days of evaluations, the remaining suspension (15 mL) containing M. enterolobii and EPNs was inoculated into Rutgers tomato seedlings. The suspension contained approximately in 300 eggs of M. enterolobii occasional juveniles and 600 IJ of each nematode species. Sixty days after inoculation were evaluated gall indexes, egg mass indexes, total number of eggs and juveniles of M. enterolobii and reproductive factor was calculated. In the mortality experiment, 500 infective juveniles of M. enterolobii and 1.000 juveniles of each EPN species were placed in a plastic pot totaling 25 mL of suspension. The evaluation of juvenile mortality was performed by counting of the mobile and immotile nematodes, by adding two drops of NaOH to the nematode suspension. It was verified that on the 10th day all ENPs provided reduction in the hatching of M. enterolobii. In the pot experiment it was found thato gall index, egg mass indexm, nematodes total number and reproduction factor were significantly reduced in treatments with all species of EPNs tested. However, in the mortality test, only EPNs S. brazilense and S. rarum provided mortality on the second day and H. bacteriophora affected mortality on the 4th day. In the other evaluations, there was no statistical difference. The results highlight the potential of the use of EPNs in programs of integrated management of M. enterolobii in tomato.

Targeted Doxorubicin Delivery to Brain Tumors via Minicells: Proof of Principle Using Dogs with Spontaneously Occurring Tumors as a Model.

BACKGROUND: Cytotoxic chemotherapy can be very effective for the treatment of cancer but toxicity on normal tissues often limits patient tolerance and often causes long-term adverse effects. The objective of this study was to assist in the preclinical development of using modified, non-living bacterially-derived minicells to deliver the potent chemotherapeutic doxorubicin via epidermal growth factor receptor (EGFR) targeting. Specifically, this study sought to evaluate the safety and efficacy of EGFR targeted, doxorubicin loaded minicells (designated EGFRminicellsDox) to deliver doxorubicin to spontaneous brain tumors in 17 companion dogs; a comparative oncology model of human brain cancers. METHODOLOGY/PRINCIPLE FINDINGS: EGFRminicellsDox were administered weekly via intravenous injection to 17 dogs with late-stage brain cancers. Biodistribution was assessed using single-photon emission computed tomography (SPECT) and magnetic resonance imaging (MRI). Anti-tumor response was determined using MRI, and blood samples were subject to toxicology (hematology, biochemistry) and inflammatory marker analysis. Targeted, doxorubicin-loaded minicells rapidly localized to the core of brain tumors. Complete resolution or marked tumor regression (>90% reduction in tumor volume) were observed in 23.53% of the cohort, with lasting anti-tumor responses characterized by remission in three dogs for more than two years. The median overall survival was 264 days (range 49 to 973). No adverse clinical, hematological or biochemical effects were observed with repeated administration of EGFRminicellsDox (30 to 98 doses administered in 10 of the 17 dogs). CONCLUSIONS/SIGNIFICANCE: Targeted minicells loaded with doxorubicin were safely administered to dogs with late stage brain cancer and clinical activity was observed. These findings demonstrate the strong potential for clinical applications of targeted, doxorubicin-loaded minicells for the effective treatment of patients with brain cancer. On this basis, we have designed a Phase 1 clinical study of EGFR-targeted, doxorubicin-loaded minicells for effective treatment of human patients with recurrent glioblastoma.

A novel serogenetic approach determines the community prevalence of celiac disease and informs improved diagnostic pathways.

BACKGROUND: Changing perspectives on the natural history of celiac disease (CD), new serology and genetic tests, and amended histological criteria for diagnosis cast doubt on past prevalence estimates for CD. We set out to establish a more accurate prevalence estimate for CD using a novel serogenetic approach. METHODS: The human leukocyte antigen (HLA)-DQ genotype was determined in 356 patients with 'biopsy-confirmed' CD, and in two age-stratified, randomly selected community cohorts of 1,390 women and 1,158 men. Sera were screened for CD-specific serology. RESULTS: Only five 'biopsy-confirmed' patients with CD did not possess the susceptibility alleles HLA-DQ2.5, DQ8, or DQ2.2, and four of these were misdiagnoses. HLA-DQ2.5, DQ8, or DQ2.2 was present in 56% of all women and men in the community cohorts. Transglutaminase (TG)-2 IgA and composite TG2/deamidated gliadin peptide (DGP) IgA/IgG were abnormal in 4.6% and 5.6%, respectively, of the community women and 6.9% and 6.9%, respectively, of the community men, but in the screen-positive group, only 71% and 75%, respectively, of women and 65% and 63%, respectively, of men possessed HLA-DQ2.5, DQ8, or DQ2.2. Medical review was possible for 41% of seropositive women and 50% of seropositive men, and led to biopsy-confirmed CD in 10 women (0.7%) and 6 men (0.5%), but based on relative risk for HLA-DQ2.5, DQ8, or DQ2.2 in all TG2 IgA or TG2/DGP IgA/IgG screen-positive subjects, CD affected 1.3% or 1.9%, respectively, of females and 1.3% or 1.2%, respectively, of men. Serogenetic data from these community cohorts indicated that testing screen positives for HLA-DQ, or carrying out HLA-DQ and further serology, could have reduced unnecessary gastroscopies due to false-positive serology by at least 40% and by over 70%, respectively. CONCLUSIONS: Screening with TG2 IgA serology and requiring biopsy confirmation caused the community prevalence of CD to be substantially underestimated. Testing for HLA-DQ genes and confirmatory serology could reduce the numbers of unnecessary gastroscopies.