New thinking for filth fly control: residual, non-chemical wall spray from volcanic glass.

Filth flies are of medical and veterinary importance because of the transfer of disease organisms to animals and humans. The traditional control methods include the use of chemical insecticides. A novel mechanical insecticide made from volcanic glass and originally developed to control mosquitoes (Imergard™ WP; ImG) was investigated for control of adult grey flesh flies, Sarcophaga bullata (Parker), secondary screwworms, Cochliomyia macellaria (F.), and house flies, Musca domestica L. In a modified WHO cone test device, the time to 50% mortality (LT50 ) when applied at 5 g/m2 (tested at 30 °C and 50% relative humidity (rH)) was 7.1, 4.3 and 3.2 h, respectively. When knockdown was included, the LT50 s were 5.5, 1.5 and 2.8 h, respectively. Application rates of 1.25 and greater g/m2 had the shortest LT50 s. The time to the LT50 increased for M. domestica as rH increased, but ImG was still active at the highest rH tested of 70%. Scanning electron micrographs showed ImG was present on all body parts, unlike that for mosquitoes where it was found mostly on the lower legs. These first studies on the use of Imergard WP against flies suggest this could be an alternative method for filth fly control.

A Canadian perspective on the use of immunoglobulin therapy to reduce infectious complications in chronic lymphocytic leukemia.

Infections are a major cause of morbidity and mortality in patients with chronic lymphocytic leukemia (cll), who typically have increased susceptibility because of hypogammaglobulinemia (hgg) related to their disease and its treatment. Immunoglobulin replacement therapy (igrt) has been shown to reduce the frequency of bacterial infections and associated hospitalizations in patients with hgg or a history of infection, or both. However, use of igrt in cll is contentious. Studies examining such treatment were conducted largely before the use of newer chemoimmunotherapies, which can extend lifespan, but do not correct the hgg inherent to the disease. Thus, the utility of igrt has to be re-evaluated in the current setting. Here, we discuss the evidence for the use of igrt in cll and provide a practical approach to its use in the prevention and management of infections.

An open-label expanded-access trial of bendamustine in patients with rituximab-refractory indolent non-Hodgkin lymphoma or previously untreated chronic lymphocytic leukemia: BEND-ACT.

BACKGROUND: Bendamustine is a bifunctional alkylating agent with unique properties that distinguish it from other agents in its class. Bendamustine is used as monotherapy or in combination with other agents to treat patients with non-Hodgkin lymphoma (nhl) and chronic lymphocytic leukemia (cll). METHODS: The prospective interventional open-label bend-act trial evaluated bendamustine in patients with rituximab-refractory indolent nhl (inhl) and previously untreated cll. Study objectives were to assess the safety and tolerability of bendamustine monotherapy and to provide patients with access to bendamustine before Health Canada approval. The study aimed to enrol up to 100 patients. All patients with inhl received an intravenous dose of bendamustine 120 mg/m(2) over 60 minutes on days 1 and 2 for up to eight 21- or 28-day treatment cycles. All patients with cll received an intravenous dose of bendamustine 100 mg/m(2) over 30 minutes on days 1 and 2 for up to six 28-day treatment cycles. RESULTS: Of 90 patients treated on study (16 with cll and 74 with inhl), 35 completed the study (4 with cll and 31 with inhl). The most common treatment-emergent adverse events (teaes) were nausea (70%), fatigue (57%), vomiting (40%), and diarrhea (33%)-mostly grades 1 and 2. Ondansetron was the most common supportive medication used in the patients (63.5% of those with inhl and 68.8% of those with cll). Neutropenia (32%), anemia (23%), and thrombocytopenia (21%) were the most frequent hematologic teaes, with neutropenia being the most common grade 3 or 4 teae leading to dose modification. Dose delays occurred in 28 patients (31.3%) because of grade 3 or 4 teaes, with a higher incidence of dose delays being observed in inhl patients on the 21-day treatment cycle than in those on the 28-day treatment cycle (50.0% vs. 24.1%). During the study, 33 patients (36.7%) experienced at least 1 serious adverse event, and 4 deaths were reported (all in patients with inhl). CONCLUSIONS: The type and frequency of the teaes reported accorded with observations in earlier clinical trials and post-marketing experiences, thus confirming the acceptable and manageable safety profile of bendamustine.

Band gap and electronic structure of an epitaxial, semiconducting Cr0.80Al0.20 thin film.

Cr(1-x)Al(x) exhibits semiconducting behavior for x = 0.15-0.26. This Letter uses hard x-ray photoemission spectroscopy and density functional theory to further understand the semiconducting behavior. Photoemission measurements of an epitaxial Cr(0.80)Al(0.20) thin film show several features in the valence band region, including a gap at the Fermi energy (E(F)) for which the valence band edge is 95 ± 14 meV below E(F). Theory agrees well with the valence band measurements, and shows an incomplete gap at E(F) due to the hole band at M shifting almost below E(F).

Evaluation of subcutaneous rituximab administration on Canadian systemic therapy suites.

Background: Non-Hodgkin lymphoma (nhl) is the most common hematologic malignancy. Diffuse large B-cell lymphoma (dlbcl) and follicular lymphoma (fl) constitute 55% of new nhl cases and are initially treated with rituximab-based chemoimmunotherapy. Relative to intravenous (IV) rituximab, a subcutaneous (sc) formulation approved in 2016 has comparable pharmacokinetics, efficacy, and safety, and a greatly reduced administration time; it is also preferred by patients. The objective of the present study was to estimate the effect (on systemic therapy suite time and on the costs of drug acquisition and administration) of implementing sc rituximab in the initial chemoimmunotherapy for fl and dlbcl over 3 years in the Canadian market. Methods: An Excel (Microsoft Corporation, Redmond, WA, U.S.A.)-based model was created with a population size based on epidemiologic data and current rituximab use, duration of use considering initial therapy, time savings for sc rituximab administration from published studies, costs from standard Canadian sources, and assumed uptake in implementing provinces of 65%, 75%, and 80% over 3 years. Key parameters and sensitivity analysis values were validated by clinical experts located in various Canadian jurisdictions. Costs are reported in 2017 Canadian dollars from the perspective of the health care system. Results: More than 3 years after implementation of sc rituximab, we estimated that 5762 Canadians would be receiving sc rituximab, resulting in savings of 128,715 hours in systemic therapy suite time and approximately $40 million in drug and administration costs. Sensitivity analyses suggest that the model is most sensitive to sc market uptake, number of induction therapy cycles, and eligible patients. Conclusions: Subcutaneous administration of rituximab can significantly reduce systemic therapy suite time and achieve substantial savings in drug and administration costs.

Viral purging of haematological autografts: should we sneeze on the graft?

High-dose cytotoxic chemotherapy followed by autologous haematopoietic stem cell transplantation (ASCT) is extensively used for the treatment of many haematopoietic, as well as several epithelial cancers. Disease relapse may be the result of tumour contamination within autograft as evidenced by gene marking studies. The multiple purging strategies that have been described to date have not proven effective in most ASCT settings. This review addresses the possibility of using oncolytic viruses as a novel purging strategy. DNA viruses such as genetically engineered adenoviral vectors have widely been used to deliver either a prodrug-activating enzyme or express wild-type p53 selectively in tumour cells in ex vivo purging protocols. In addition, conditionally replicating adenoviruses that selectively replicate in tumour cells and herpes simplex virus type 1 are other DNA viruses that have been tested as ex vivo purging agents under laboratory conditions. Vesicular stomatitis virus (VSV) and reovirus are naturally occurring RNA viruses that appear to hold promise as purging agents under ex vivo and in vivo settings. Preclinical data demonstrate reovirus's purging potential against breast, monocytic and myeloma cell lines as well as patient-derived tumours of diffuse large B-cell lymphoma, chronic lymphocytic leukaemia, Waldenstrom macroglobulinemia and small lymphocytic lymphoma. In addition, VSV has shown effective killing of leukaemic cell lines and multiple myeloma patient specimens. Given the increasing interest in the utilization of viruses as purging agents, the following review provides a timely summary of the potential and the challenges of oncolytic viruses as purging modalities during ASCT.

Functional analysis of a duplicated linked pair of ribosomal protein genes in Saccharomyces cerevisiae.

Ribosomal protein genes RP28 and S16A (RP55) are closely linked. Another set of this pair of genes exists in the genome (copy 2), genetically unlinked to copy 1. By using gene replacement techniques, we have shown that RP28 from copy 1 is required for vegetative growth and that the cells need S16A from copy 2 to achieve maximum growth rate.

Prohibitin, an evolutionarily conserved intracellular protein that blocks DNA synthesis in normal fibroblasts and HeLa cells.

Genes that act inside the cell to negatively regulate proliferation are of great interest because of their implications for such processes as development and cancer, but these genes have been difficult to clone. This report details the cloning and analysis of cDNA for prohibitin, a novel mammalian antiproliferative protein. Microinjection of synthetic prohibitin mRNA blocks entry into S phase in both normal fibroblasts and HeLa cells. Microinjection of an antisense oligonucleotide stimulates entry into S phase. By sequence comparison, the prohibitin gene appears to be the mammalian analog of Cc, a Drosophila gene that is vital for normal development.

Overnight storage of autologous stem cell apheresis products before cryopreservation does not adversely impact early or long-term engraftment following transplantation.

To reduce costs and avoid inconvenient overtime work, our institution changed policy in September 2000 so that autologous stem cell apheresis products were stored overnight before cryopreservation rather than immediately processed. This retrospective review was conducted to evaluate the possible impact of this policy change on hematopoietic engraftment following autologous stem cell transplantation (ASCT). In total, 229 consecutive lymphoma patients who underwent a single, unpurged ASCT in Calgary between January 1995 and November 2003 were evaluated. Of these patients, 131 patients' autografts underwent immediate processing and cryopreservation before September 2000, and 98 patients' autografts underwent next-day cryopreservation after overnight storage following this date. Results of univariate and multivariate analyses demonstrated no adverse effect of overnight storage before cryopreservation on the number of days to initial engraftment of platelets or neutrophils, on the proportion of patients with low blood counts 6 months post-ASCT, or on lymphoma relapse rates or overall survival post-ASCT. These data suggest that overnight storage of the autograft before cryopreservation does not adversely affect graft viability or influence long-term disease status, and support the continued use of overnight storage of stem cells before cryopreservation as a convenient, cost reduction measure.

Psoralen-modified clamp-forming antisense oligonucleotides reduce cellular c-Myc protein expression and B16-F0 proliferation.

The c-myc protooncogene plays an important role in the abnormal growth pattern of melanoma cells. In an attempt to inhibit c-Myc expression and the growth of an established murine melanoma cell line, we targeted homopurine sequences within the mouse myc mRNA with modified antisense oligonucleotides (AS ODNs). Psoralen was conjugated to the 5'-end of these clamp-forming oligonucleotides (clamp ODNs). Gel mobility shift analysis demonstrated a sequence-specific interaction between the active clamp ODNs (Myc-E2C and Myc-E3C) and the 1.4 kb c-myc mRNA, but no interaction with the control clamp ODN (SCR**). This association was further confirmed by thermal denaturation studies. In vitro translation assays demonstrated that both Myc-E2C and Myc-E3C at 5 microM inhibited c-Myc expression >99% after UV activation at 366 nm. Immunostaining of B16-F0 cells with a c-Myc monoclonal antibody revealed a significant reduction in c-Myc after clamp ODN treatment compared with the untreated or SCR** control-treated cells. This result was corroborated by western blot analysis. Utilizing the MTT assay to determine the effects of ODN-mediated c-Myc reduction on B16-F0 growth, we observed 60 and 64% reductions in growth after treatment with 5 microM Myc-E3C and Myc-E2C, respectively. We attribute the enhanced effectiveness of the clamp ODNs to psoralen activation. Our preliminary data suggest that inhibiting c-Myc overexpression results in a significant reduction in abnormal proliferation of B16-F0 melanoma cells and that the increased efficiency of clamp ODNs may provide an important advantage for their use in antisense therapies.

Autologous transplantation for primary systemic AL amyloidosis is feasible outside a major amyloidosis referral centre: the Calgary BMT Program experience.

Recent reports from large amyloidosis referral centers suggest that primary systemic AL amyloidosis patients treated with high-dose melphalan (HDM) and autologous stem cell transplantation (ASCT) survive longer than historical controls treated with less intensive chemotherapy, despite high transplant-related mortality (TRM) rates of >10%. A retrospective review was conducted to determine if the outcome of ASCT for AL amyloidosis at our institution was similar to that reported at major amyloidosis referral centers. Over a 7 year period, we treated a total of 15 AL amyloidosis patients with ASCT, including four with poor prognosis cardiac or multisystem involvement. No TRM was observed. Overall, 10 patients (67%) achieved a complete hematological response and four patients (27%) achieved a complete organ response. The 4-year event-free and overall survival rates were 60% (95% CI 32-89%) and 75% (95% CI 50-100%), respectively. One patient, who presented with cardiac failure and multiorgan involvement with colonic bleeding currently remains in complete remission 62 months post-ASCT. In conclusion, ASCT for primary AL amyloidosis can safely be performed at experienced transplant centers that are not associated with major amyloidosis referral centers, and is feasible for patients who have multisystem involvement, particularly for motivated patients with good performance status.

Biological purging of breast cancer cell lines using a replication-competent oncolytic virus in human stem cell autografts.

Autologous hematological stem cell transplantation (ASCT) is used for the treatment of many hematological and several solid cancers. ASCT, however, has proven disappointing as a therapeutic strategy for breast cancer. Our group and others have previously shown that breast cancer micrometastases found in patients' apheresis products (APs) predict shorter progression-free and overall survival. The implications of this finding are twofold: (i) contaminating tumor cells (CTCs) in AP reflect a higher systemic disease burden and/or (ii) reinfused CTCs contribute to relapse/progressive disease. To date, purging strategies have been disappointing. We have previously demonstrated the oncolytic properties of reovirus in in vitro, in vivo and ex vivo systems. In the present study, we tested the hypothesis that reovirus purges CTCs in a breast cancer cell line purging model. Reovirus-infected human breast cancer cell lines (HTB 133, HTB 132, SKBR3 and MCF7) exhibited cell death within days. Admixtures of AP with cells from breast tumor cell lines, which were then exposed to reovirus, showed complete purging of CTCs (assessed via flow cytometry/tumor cell outgrowth analysis) without deleterious effect on CD34+ cells. Our results provide preclinical support for the ex vivo use of reovirus as a purging modality for breast cancer during ASCT.

Granulocyte progenitor cell (CFUC) harvest by continuous apheresis in dogs. Effects of blood volume and lithium on yields.

The ability to harvest large amounts of hematopoietic stem cells from blood would eliminate the more difficult approach of bone marrow harvest. Unfortunately, concentration of stem cells in the blood compartment is less than 1% of their concentration in bone marrow. Attempts to increase harvest of blood stem cells, as assayed by granulocyte progenitor cells (CFUC), have been only partially successful. Our study confirms previous reports that CFUC can be mobilized into the blood compartment in dogs, but this mobilization is rate-limited. Unlike platelets and granulocytes that are effectively harvested during the first blood volume processed by continuous apheresis, effective CFUC harvest begins during the second blood volume (606 +/- 97.9 CFUC/ml), peaks by the third (740 +/- 30 CFUC/ml), and remains constant through five blood volumes processed (700 +/- 272 CFUC/ml). Since blood CFUC concentration falls at the end of five blood volumes processed (40% of initial values), further continuous apheresis would not be effective. Treatment of animals with lithium did not improve CFUC harvest. These results show that apheresis procedures can be developed to a limited extent to increase the harvest of hematopoietic progenitor and stem cells.

Antiangiogenic gene therapy for cancer via systemic administration of adenoviral vectors expressing secretable endostatin.

A growing number of antiangiogenesis strategies have been investigated for the treatment of cancer and other angiogenesis-dependent diseases. One of the most promising strategies is to systemically administer one or more antiangiogenic proteins frequently enough to achieve a sufficient long-term steady state level of the protein(s) to achieve the maximum beneficial effect. However, the utility of this strategy is limited because of many technical difficulties, including obtaining both the quantity and quality of the protein(s) necessary for optimal therapeutic benefit. To overcome these difficulties, we hypothesized that a single administration of a replication-defective adenoviral vector expressing a secretable antiangiogenic protein could achieve an optimal long-term systemic concentration. We constructed a recombinant adenoviral vector, Av3mEndo, which encodes a secretable form of murine endostatin. We demonstrated secretion of endostatin from several cell lines transduced with Av3mEndo. Partially purified endostatin secreted from Av3mEndo-transduced mammalian cells was shown to potently inhibit endothelial cell migration in vitro. A single intravenous administration of Av3mEndo in mice was shown to result in (1) prolonged and elevated levels of circulating endostatin, (2) partial inhibition of VEGF-induced angiogenesis in a VEGF implant angiogenesis model, and (3) prolonged survival and in 25% of mice the complete prevention of tumor growth in a prophylactic human colon/liver metastasis xenograft murine model. These results support our contention that adenoviral vector-mediated expression of an antiangiogenic protein(s) represents an attractive therapeutic approach to cancer and other angiogenesis-dependent diseases.

Discordant effects of aging on prolactin and luteinizing hormone-beta messenger ribonucleic acid levels in the female rat.

To examine the molecular genetic basis for the age-related increase in PRL secretion and decrease in LH production in the rat, we measured steady state levels of PRL and LH beta mRNA in pituitary homogenates and cell lysates from monolayer adenohypophyseal cultures. These mRNA levels were compared with the corresponding levels of immunoreactive PRL and LH in sera and culture media. Paired groups (n = 4-10/group) of intact and 4-week ovariectomized mature (6-7 months old) and old (23-25 months old) female Wistar rats were studied. Serum PRL levels were 550% higher in intact old vs. mature rats (P less than 0.001), whereas the corresponding pituitary homogenate levels of PRL mRNA were similar (P greater than 0.4). Medium PRL concentrations were 230% greater (P less than 0.006) whereas cell lysate concentrations of PRL mRNA were unaltered (P greater than 0.2) in monolayer cultures from intact old vs. mature rats. Serum PRL levels were 650% higher (P less than 0.003) and pituitary homogenate PRL mRNA levels were slightly increased (P less than 0.04) in ovariectomized old vs. mature rats. Neither serum LH values (P greater than 0.07) nor pituitary homogenate LH beta mRNA levels (P greater than 0.1) differed in intact old and mature rats, whereas the corresponding medium concentrations of LH were reduced (P less than 0.001). Ovariectomized old vs. mature rats exhibited reductions in serum (P less than 0.02) and medium (P less than 0.001) LH concentrations, as well as in pituitary homogenate (P less than 0.002) and cell lysate (P less than 0.006) LH beta mRNA levels. Thus, these data revealed coordinate decreases with age in LH beta mRNA and LH secretion, particularly in ovariectomized rats, suggesting an age-related alteration at or before LH beta gene transcription. These findings parallel observations on other genes whose products change with age. In contrast, the observation that the increased secretion of PRL in old rats is accompanied by little or no increase in PRL mRNA is novel and suggests that age-related alterations in PRL gene expression proceed through a posttranscriptional mechanism.

Regions of evolutionary conservation between the rat and human prohibitin-encoding genes.

We have analyzed and compared the 5' promoter region, the intron structure and the exon-intron flanking sequences in the rat and human prohibitin-encoding genes (PHB). Comparative analysis of a 350-nt region immediately 5' to and including the first exon identifies eight highly conserved regions, four of which correspond to binding sites for known transcriptional control proteins (CCAAT box, 'SV40' site and two Sp1 sites). The promoter lacks a TATA box. Four transcription start points (tsp) clustered within a 35-bp region were identified by rapid amplification of cDNA ends (RACE). The exon-intron boundaries in rat and human are highly conserved, with identical positioning of splice junctions. PCR analysis with conserved exon primers was used to detect length variation between rat and human PHB, and length differences were observed in all of the introns.

Studies into the transplantation biology of ultraviolet light-induced tumors.

The majority of skin tumors induced in mice by ultraviolet (UV) light are rejected when implanted into normal syngeneic recipients. Subcarcinogenic levels of UV light exposure render the normally resistant mice susceptible to tumor challenge. The immunoregulatory effect of UV light appears to be additive, since the growth rate of a tumor transplant is dependent upon the length of UV exposure administered prior to implantation. This suppressive influence does not appear to be directly mediated by the UV light, because the amputation of UV-irradiated tail skin allows for a retention of tumor resistance in otherwise tumor-susceptible hosts. UV-irradiated mice could also be immunized against UV tumors, which suggests that immune recognition of tumor-specific transplantation antigens has not been inhibited. The ability of UV exposure to alter normal immunological reactivity to UV-induced tumors is possibly an integral factor in the mechanism underlying UV carcinogenesis.

Early development of children with sex chromosome aberrations.

A prospective study was made of 42 children who were found at birth to have sex chromosome aberrations. The mean developmental quotient of the XXY and XXX groups was in the dull-normal range, whereas developmental quotients of 3 XYY children were low normal. The commonest deficits were in language and gross motor skills, with highest incidence in the XXX group. Many of the children had behavior problems, but no specific pattern was associated with an individual chromosome aberration. The XXX group experienced a linear growth spurt after 4 years of age. With one exception there were only mild dysmorphic features, none of them specifically associated with any subgroup. Of 17 children followed up to school age, almost two thirds evidenced learning and/or behavior problems. There was a high incidence of birth problems particularly in XXX children (seven of ten), and of social problems, psychiatric illness and learning difficulties in their families. In 14 cases, the parent-child relationship was disturbed, resulting in behavior problems. These findings make it improbable that the children's generally poor developmental and behavioral performance were solely attributable to their chromosomal constitution.

Acute withdrawal of nasal CPAP in obstructive sleep apnea does not cause a rise in stress hormones.

We hypothesized that withdrawal of nasal continuous positive airway pressure (CPAP) in patients with sleep apnea would produce a measurable stress response. To test this hypothesis, we ceased CPAP in eight patients regularly using nasal CPAP long term and measured the effect on sleep apnea as well as plasma and urinary levels of the stress hormones, noradrenaline, cortisol and adrenocorticotropic hormone (ACTH). CPAP withdrawal led to an immediate recurrence of sleep apnea with increases in apnea index, arousal index and oxygen desaturation (all p < .0001) but no change in levels of noradrenaline, cortisol or ACTH. We conclude that acute withdrawal of CPAP in patients with sleep apnea does not lead to a classic stress response.

Myocardial protection during ischemic cardiac arrest. The importance of magnesium in cardioplegic infusates.

The increasing use of coronary infusates for the protection of the human heart drug ischemic cardiac arrest has placed great emphasis on the need for a rational and safe formulation of any infusion solution. Using a rat heart model of cardiopulmonary bypass and ischemic cardiac arrest, we have found magnesium to be a highly effective component of protective infusates which can be additive to hypothermia and other protective agents. However, the concentration of magnesium bears a complex relationship to the degree of protection, a fact which stresses the need for the establishment of the correct concentration for optimal protection.