Costs of fracture-related infection: the impact on direct hospital costs and healthcare utilisation.

PURPOSE: Fracture-Related Infection (FRI) is associated with high medical costs and prolonged healthcare utilization. However, limited data is available on the financial impact. The purpose of this study was to investigate the impact of FRI on direct hospital costs and healthcare utilization. METHODS: This was a retrospective cohort study in a level-1 trauma centre in the Netherlands. Patients ≥ 18 years, after open reduction and internal fixation of a long bone fracture between January 1st 2016 and November 1st 2021, were included. Exclusion criteria were Injury Severity Score (ISS) ≥ 16, indefinable data on costs or incomplete follow-up. Hospital costs related to fracture treatment were individually calculated based on procedure codes raised with a fixed percentage of overhead expenses, in line with hospital billing policies. RESULTS: In total, 246 patients were included with a median follow-up of 1 year (IQR 0.6-1.8). A total of 45 patients developed FRI, of whom 15 patients had an FRI recurrence. Compared to non-FRI patients, median hospital costs from an FRI patient without and with recurrence, were respectively three (3.1) and seven (7.6) times higher. Compared to non-FRI patients, increased costs in patients with FRI or recurrent FRI are due to respectively a fivefold or even tenfold prolonged length-of-stay, two or seven additional infection-related surgeries, and 21 or 55 days of intravenous antibiotic treatment. CONCLUSION: Direct healthcare costs of patients with single occurrence of FRI after long bone fracture treatment are three times higher compared to non-FRI patients. In case of FRI-recurrence, the differences in costs might even increase to sevenfold. To put this in perspective, cost of severely injured trauma patients were recently established at approximately 25.000 euros. Compared to non-FRI patients, increased costs in patients with FRI or recurrent FRI are due to respectively a fivefold or even tenfold prolonged length-of-stay, two or seven additional infection-related surgeries and 21 or 55 days of intravenous antibiotic treatment. Not only from patient perspective but also from a financial aspect, it is important to focus on prevention of (recurrent) FRI.

Eosinophilic esophagitis prevalence, incidence, and presenting features: a 22-year population-based observational study from southwest Sweden.

Eosinophilic esophagitis (EoE) is a chronic inflammatory condition of the esophagus that affects both children and adults. Symptoms in adults are mainly esophageal dysphagia, which ranges from mild symptoms to acute food bolus obstruction of the esophagus. Diagnosis is defined as symptoms of esophageal dysfunction and ≥ 15 eosinophils/high power field (HPF) in at least one of the biopsies taken from the esophagus. EoE appears to be increasing in both prevalence and incidence. The aim of this study was to investigate the prevalence, incidence, and presenting symptoms of patients with EoE within the catchment area of Northern Älvsborg County Hospital in Trollhättan. Patient records with the ICD code of EoE between 2012 and 2022 and pathology reports from esophageal biopsies from 2000-2022 were examined. Patients with symptoms of esophageal dysfunction and > 15 eosinophils/HPF were classified as having EoE. In total, 409 EoE patients (379 adults and 30 children) fulfilled the diagnostic criteria during the follow-up period. The overall prevalence was 113 cases/100 000 inhabitants (adults 127/100 000 and children 57/100 000) at 31 December 2022. The incidence was 7/100 000 and increased during the observation period. At diagnosis, 46% of the adults and 11% of the children had a history of acute bolus obstruction requiring hospitalization, while 51% of adults and 22% of children exhibited endoscopic findings of fibrosis. The prevalence of EoE is significantly higher than that generally reported in an area of southwest Sweden. The results indicate that the incidence is increasing; however, whether this is due to an actual increase or heightened awareness of EoE is inconclusive. Acute bolus obstruction is a common presenting symptom among EoE patients and is most likely an effect of late diagnosis.

The QUILT study: quilting sutures in patients undergoing breast cancer surgery: a stepped wedge cluster randomized trial study.

BACKGROUND: Seroma is the most common complication following breast cancer surgery, with reported incidence up to 90%. Seroma causes patient discomfort, is associated with surgical site infections (SSI), often requires treatment and increases healthcare consumption. The quilting suture technique, in which the skin flaps are sutured to the pectoralis muscle, leads to a significant reduction of seroma with a decrease in the number of aspirations and surgical site infections. However, implementation is lagging due to unknown side effects, increase in operation time and cost effectiveness. Main objective of this study is to assess the impact of large scale implementation of the quilting suture technique in patients undergoing mastectomy and/or axillary lymph node dissection (ALND). METHODS: The QUILT study is a stepped wedge design study performed among nine teaching hospitals in the Netherlands. The study consists of nine steps, with each step one hospital will implement the quilting suture technique. Allocation of the order of implementation will be randomization-based. Primary outcome is 'textbook outcome', i.e.no wound complications, no re-admission, re-operation or unscheduled visit to the outpatient clinic and no increased use of postoperative analgesics. A total of 113 patients is required based on a sample size calculation. Secondary outcomes are shoulder function, cosmetic outcome, satisfaction with thoracic wall and health care consumption. Follow-up lasts for 6 months. DISCUSSION: This will be one of the first multicentre prospective studies in which quilting without postoperative wound drain is compared with conventional wound closure. We hypothesize that quilting is a simple technique to increase textbook outcome, enhance patient comfort and reduce health care consumption.

Diagnostic accuracy of serum inflammatory markers in late fracture-related infection: a systematic review and meta-analysis.

AIMS: To assess the diagnostic value of C-reactive protein (CRP), leucocyte count (LC), and erythrocyte sedimentation rate (ESR) in late fracture-related infection (FRI). MATERIALS AND METHODS: PubMed, Embase, and Cochrane databases were searched focusing on the diagnostic value of CRP, LC, and ESR in late FRI. Sensitivity and specificity combinations were extracted for each marker. Average estimates were obtained using bivariate mixed effects models. RESULTS: A total of 8284 articles were identified but only six were suitable for inclusion. Sensitivity of CRP ranged from 60.0% to 100.0% and specificity from 34.3% to 85.7% in all publications considered. Five articles were pooled for meta-analysis, showing a sensitivity and specificity of 77.0% and 67.9%, respectively. For LC, this was 22.9% to 72.6%, and 73.5% to 85.7%, respectively, in five articles. Four articles were pooled for meta-analysis, resulting in a 51.7% sensitivity and 67.1% specificity. For ESR, sensitivity and specificity ranged from 37.1% to 100.0% and 59.0% to 85.0%, respectively, in five articles. Three articles were pooled in meta-analysis, showing a 45.1% sensitivity and 79.3% specificity. Four articles analyzed the value of combined inflammatory markers, reporting an increased diagnostic accuracy. These results could not be pooled due to heterogeneity. CONCLUSION: The serum inflammatory markers CRP, LC, and ESR are insufficiently accurate to diagnose late FRI, but they may be used as a suggestive sign in its diagnosis.

A systematic approach to predicting the risk of unicompartmental knee arthroplasty revision.

OBJECTIVE: Unicompartmental knee arthroplasty (UKA) revision is usually due to the degenerative degree of knee articular osteochondral tissue in the untreated compartment. However, it is difficult to simulate the biomechanical behavior on this tissue accurately. This study presents and validates a reliable system to predict which osteoarthritis (OA) patients may suffer revision as a result of biomechanical reasons after having UKA. DESIGN: We collected all revision cases available (n = 11) and randomly selected 67 UKA cases to keep the revision prevalence of almost 14%. All these 78 cases have been followed at least 2 years. An elastic model is designed to characterize the biomechanical behavior of the articular osteochondral tissue for each patient. After calculated the force on the tissue, finite element method (FEM) is applied to calculating the strain of each tissue node. Kernel Ridge Regression (KRR) method is used to model the relationship between the strain information and the risk of revision. Therefore, the risk of UKA revision can be predicted by this integrated model. RESULTS: Leave-one-out (LOO) cross-validation (CV) is implemented to assess the prediction accuracy. As a result, the mean prediction accuracy is 93.58% for all these cases, demonstrating the high value of this model as a decision-making assistant for surgical plaining of knee OA. CONCLUSIONS: The results of this study demonstrated that this integrated model can predict the risk of UKA revision with theoretically high accuracy. It combines bio-mechanical and statistical learning approach to create a surgical planning tool which may support clinical decision in the future.

Impact of straw mulch on populations of onion thrips (Thysanoptera: Thripidae) in onion.

Development of insecticide resistance in onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae), populations in onion (Allium spp.) fields and the incidence of the T. tabaci transmitted Iris yellow spot virus have stimulated interest in evaluating alternative management tactics. Effects of straw mulch applied in commercial onion fields in muck areas of western New York were assessed in 2006 and 2007 as a possible onion thrips management strategy. In trials in which no insecticides were applied for thrips control, straw mulch-treated plots supported significantly lower T. tabaci populations compared with control plots. In both years, the action thresholds of one or three larvae per leaf were reached in straw mulch treatments between 7 and 14 d later than in the control. Ground predatory fauna, as evaluated by pitfall trapping, was not increased by straw mulch in 2006; however, populations of the common predatory thrips Aeolothrips fasciatus (L.) (Thysanoptera: Aeolothripidae) were significantly lower in straw mulch plots in both years. Interference of straw mulch in the pupation and emergence of T. tabaci was investigated in the lab and their emergence was reduced by 54% compared with bare soil. In the field the overall yield of onions was not affected by the straw mulch treatment; however, the presence of jumbo grade onions (>77 mm) was increased in 2006, but not in 2007. These results indicate that populations of T. tabaci adults and larvae can be significantly reduced by the use of straw mulch without compromising overall onion yield. The use of this cultural practice in an onion integrated pest management program seems promising.

Advances in control of onion thrips (Thysanoptera: Thripidae) in cabbage.

Onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae), feeding injury results in discoloration and a rough texture on cabbage, Brassica oleracea capitata (L.), leaves, and damage may occur deep inside the head. It has become a key pest of cabbage in the United States and many other countries. Previous studies have indicated poor control using insecticides. The present study identified imidacloprid drenches and sprays of acetamiprid, dimethoate, spinosad, and imidacloprid as insecticides that performed better than the industry standard, lambda-cyhalothrin. However, additional tests with foliar sprays of dimethoate and acetamiprid indicated there was not an ideal crop stage (precupping, cupping, or postcupping) at which either insecticide could be applied for reliable control of T. tabaci, possibly because of multiple flights of thrips into the crop or the asynchrony of flights and susceptible crop stages. In tests in a commercial field, a soil drench of imidacloprid 4 wk after transplanting reduced the number of damaged leaves in the head by 32%, whereas five sprays of acetamiprid reduced damage by 51%. Combining both insecticide regimes reduced damage by 85%, but resulted in a very costly management program. Cabbage varieties varied considerably in susceptibility with some having negligible thrips injury, regardless of being treated with an insecticide. Planting date affected susceptibility of cabbage to some degree, but not as much as other tactics. Overall, these studies indicate that increased emphasis should be placed on breeding cabbages to be resistant to T. tabaci as the foundation for its management.

Overwintering locations and hosts for onion thrips (Thysanoptera: Thripidae) in the onion cropping ecosystem in New York.

Identifying locations where onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae), overwinter and subsequently disperse is important for designing control strategies. In upstate New York from 2003 through 2006, potential overwintering sites in the commercial onion, Allium cepa L., cropping system were investigated early in the spring before onion seedling emergence and again late in the season after onions were harvested. Onion thrips adults were sampled directly from the soil and indirectly from the soil by using emergence cages. Sampling locations included onion field interiors and edges and areas outside of these fields, including woods. Host material sampled included onion culls; volunteer onions, which sprout from cull onions left behind after harvest; and weeds. Onion thrips adults were found in all sections of onion fields and in locations outside of onion fields, with the fewest emerging from woods. Emergence began in early May and extended into June. Peak emergence occurred during the last half of May, at which time 50-75% of the population had emerged. Adults colonized volunteer onions as early as late March and as late as mid-November. No adults were found overwintering in onion cull piles. Adults also colonized several weed species, especially pigweed, Amaranthus hybridis L., and lambsquarters, Chenopodium album L., late in the fall. Our results indicate that onion thrips adults overwinter in the soil within and near onion fields and that they probably colonize volunteer onion plants before subsequent generations infest the onion crop in the spring. Volunteer onions and weeds also provide onion thrips with a host after onions are harvested. Consequently, onion thrips management strategies should include tactics that reduce volunteer onion and weed abundance.

Patterns of insecticide resistance in onion thrips (Thysanoptera: Thripidae) in onion fields in New York.

To develop an insecticide resistance management program for onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae), on onions (Allium spp.), we surveyed populations in commercial onion fields in New York and evaluated their susceptibility to the two most widely used classes of insecticides plus two new insecticides during 2003-2005. All insecticide evaluations were conducted using the Thrips Insecticide Bioassay System (TIBS). As in our surveys conducted during 2002-2003, there were large temporal and spatial variations in susceptibility to the pyrethroid lambda-cyhalothrin (Warrior) across onion-growing regions in 2003. New data indicate that the field rate of methomyl (Lannate LV) still provides control but that the genes for resistance to methomyl are present in some populations. Tests with the two new insecticides, acetamiprid (Assail 70 WP) and spinosad (SpinTor 2CS), indicated they provided > 85% mortality at the field rate. To determine the spatial variation in insecticide susceptibility within a region, a series of systematic assays were conducted with lambda-cyhalothrin and methomyl. In 2004 and 2005, our data indicated that the within-region spatial variation in susceptibility to lambda-cyhalothrin was not large at the field rate or for the 100 ppm rate of methomyl. In 2005, a year in which T. tabaci densities in most fields were much higher than in 2004, growers were unable to control T. tabaci in particular fields and attributed this lack of control to resistance. Yet, we found similar levels of high susceptibility in all fields when using TIBS. This finding suggests that resistance had not developed and that variation in control may have been due to other factors, such as localized higher populations, poor spray coverage, too much time between spray applications, or different onion varieties.

Regional and temporal variation in susceptibility to lambda-cyhalothrin in onion thrips, Thrips tabaci (Thysanoptera: Thripidae), in onion fields in New York.

Populations of onion thrips, Thrips tabaci Lindeman, from commercial onion fields in New York were evaluated for their susceptibility to the commonly used pyrethroid, lambda-cyhalothrin (Warrior T), using a novel system called the Thrips Insecticide Bioassay System (TIBS). To use TIBS, thrips are collected directly from the plant into an insecticide-treated 0.5-ml microcentrifuge tube that has a flexible plastic cap with a small well into which 0.08 ml of a 10% sugar-water solution with food colorant is deposited. The solution is sealed into the well with a small piece of stretched parafilm through which the thrips can feed on the solution. Thrips mortality is assessed after 24 h with the help of a dissecting stereoscope. In 2001, onion thrips populations were collected from 16 different sites and resistance ratios were >1,000 in five populations. Percent mortality at 100 ppm, a recommended field rate, varied from 9 to 100%, indicating high levels of variation in susceptibility. Particular instances of resistance appeared to be the result of practices within an individual field rather than a regional phenomenon. In 2002, we also observed large differences in onion thrips susceptibility, not only between individual fields but also between thrips collected in a single field at mid season and late season, again suggesting that insecticide-use practices within an individual field caused differences in susceptibility. Additional tests indicated no differences in susceptibility between adult and larval onion thrips populations and only relatively minor differences between populations collected from different parts of the same field. Using TIBS, several populations of onion thrips with different susceptibilities to lambda-cyhalothrin were identified and then subjected to lambda-cyhalothrin-treated onion plants. There was a highly significant positive relationship between percent mortality of thrips from TIBS and percent mortality from the treated onion plants, indicating that results from TIBS could be used to predict spray performance. These data suggest that use of TIBS for evaluating susceptibility to particular insecticides could be instrumental for developing a resistance management strategy for onion thrips.

Gene expression in chronic lymphocytic leukemia B cells and changes during induction of apoptosis.

Our studies in chronic lymphocytic leukemia (CLL) are directed at understanding which signals maintain viability in vivo and become lost upon removal of leukemic cells from the body, such that they immediately begin to undergo apoptosis ex vivo. In this report, we examine changes in gene expression observed between freshly isolated CLL B cells and after maintenance in vitro with and without Fludara. We compare these effects with an Epstein-Barr virus (EBV)-transformed cell line treated similarly. Kinetic effects of drug treatment on apoptosis and cell division were examined with DNA laddering, radioisotopic labeling, and flow cytometry using the fluorescent dye carboxyfluorescein diacetate succinimidyl ester. Reverse transcriptase polymerase chain reaction and hybridization blots of microarray cDNA analyses were performed to examine gene expression. We demonstrate that many genes, especially cyclin D1, were downregulated after culture of CLL cells. Anti-apoptotic genes BAG-1 and Akt2 were upregulated. The greatest positive effect with Fludara was the upregulation of JNK1. The EBV-transformed cell line was resistant to classic DNA laddering induced with Fludara. Although DNA synthesis was blocked, the EBV-transformed cell line had some ability to recover from treatment following drug washout. CLL cells express cell cycle regulatory genes that are specific for activated cells in the G(1)-S phase of the cell cycle. Growth regulatory signals are lost when the leukemic cells are isolated from the body. Fludara enhances kinetics of apoptosis and induces expression of a gene responsive to stress that regulates expression of a kinase involved in initiation of the apoptotic pathway.

Role of beta2 integrins in the prevention of apoptosis induction in chronic lymphocytic leukemia B cells.

Immunologically committed lymphocytes, especially mature, leukemic B cells, proliferate then accumulate without further cell division in chronic lymphocytic leukemia patients (CLL). These mature, leukemic B cells often produce autoantibodies. Under normal circumstances, immunologically committed lymphocytes that are autoreactive are deleted by a programmed cell death mechanism. In CLL cells, these mechanisms appear to be inhibited; therefore, cells accumulate rather than be destroyed. To understand the mechanism by which cell survival is selected over death in CLL cells, we studied the role of beta2 integrins and their ligands in the regulation of apoptosis. CLL cells were treated with monoclonal antibodies directed against beta2 integrins. Antibodies directed against the I-domain of the alpha chain of CD11b/CD18 inhibited apoptosis. The identity of the physiological ligand or counter-receptor for beta2 integrins that was required for the inhibition of apoptosis induction was sought. The ligand iC3b, but not ICAM-1 or fibrinogen, was identified as a ligand that could prevent apoptosis of CLL B cells. Free iC3b levels were elevated in CLL patients indicating that this ligand is available in vivowhere it may interact with beta2 integrins on CLL B cells and sustain their viability by preventing activation of the programmed cell death pathway. Leukemia (2000) 14, 34-39.

Immune cell functions in pancreatic cancer.

Pancreatic cancer kills nearly 29,000 people in the United States annually-as many people as are diagnosed with the disease. Chemotherapeutic treatment is ineffective in halting progression of the disease. Yet, specific immunity to pancreatic tumor cells in subjects with pancreatic cancer has been demonstrated repeatedly during the last 24 years. Attempts to expand and enhance tumor-specific immunity with biotherapy, however, have not met with success. The question remains, "Why can't specific immunity regulate pancreatic cancer growth?" The idea that tumor cells have evolved protective mechanisms against immunity was raised years ago and has recently been revisited by a number of research laboratories. In pancreatic cancer, soluble factors produced by and for the protection of the tumor environment have been detected and are often distributed to the victim's circulatory system where they may effect a more generalized immunosuppression. Yet the nature of these soluble factors remains controversial, since some also serve as tumor antigens that are recognized by the same T cells that may become inactivated by them. Unless the problem of tumor-derived immunosuppressive products is addressed directly through basic and translational research studies, successful biotherapeutic treatment for pancreatic cancer may not be forthcoming.

Immunoregulation in pancreatic cancer patients.

Metastatic pancreatic cancer is one of the most aggressive cancer known in man yet specific antitumor immunity has been demonstrated in lymph nodes draining the sites of pancreatic tumors. Despite this immunity, pancreatic cancer patients suffer a quick demise. To further define tumor immunity in patients with metastatic pancreatic cancer, we sought to characterize helper T cell subsets, serum cytokines, cellular cytotoxicity that is both T-cell and non-T cell mediated, as well as known tumor-derived immunosuppressive products that may be present in their peripheral blood. Significantly heightened levels of interleukin 2 (IL-2), a Th1 cytokine, were found in patients before treatment with chemotherapy while serum IL-10, a Th2 cytokine, were at significantly lower levels than observed in normal donors tested between their fifth and seventh decades of life. IL-10 levels increased progressively with age as a serum-bound protein in normal, healthy donors tested between the ages of 24 through 61. An age associated progression of increased IL-10 levels was not observed in pancreatic cancer patients. Few patients had detectable serum levels of soluble fas ligand but approximately half had elevated levels of a tumor marker, detected with the CA-15.3 assay, known as soluble MUCIN 1 (MUC1). Cell mediated cytotoxicity including T-cell mediated killing of pancreatic tumor cell lines was detected in many patients. These data suggest that pancreatic cancer patients have activated type 1 helper T cells that can support development of cell-mediated immunity, and that their sera contain lowered levels of the "anti-inflammatory" type 2 cytokine, IL-10.

Mycobacterium avium complex activates nuclear factor kappaB via induction of inflammatory cytokines.

A variety of microorganisms has been reported to directly induce NF-kappaB, a critical step in the regulation of genes involved in the cellular immune response. In this study, we demonstrate that proinflammatory cytokines such as tumor necrosis factor alpha (TNFalpha) produced upon activation by the Mycobacterium avium complex (MAC) preceed NF-kappaB activity in U937, a human monocytoid cell line. MAC induction of TNFalpha mRNA expression was detected within 15 min after MAC infection, whereas enhanced NF-kappaB binding activity was not detected until 90 to 120 min postinfection. Supershift analysis revealed increased p50 in the MAC-induced NF-kappaB binding complexes. Consistent with an autocrine mechanism, anti-TNFalpha antibody and dexamethasone, a known cytokine inhibitor, both completely suppressed the effect of MAC on the induction of NF-kappaB. Taken together, these findings suggest that exposure of monocyte cell membranes to MAC induces endogenous TNFalpha, which in turn enhances NF-kappaB binding activity. The rapid induction of TNFalpha may be important in the initial host response to MAC infection.

Imbalanced expression of the glucocorticoid receptor isoforms in cultured lymphocytes from a patient with systemic glucocorticoid resistance and chronic lymphocytic leukemia.

The human glucocorticoid receptor (GR) is expressed as two alternatively spliced isoforms, GRalpha and GRbeta. Whereas GRalpha is a hormone-activated transcription factor, GRbeta does not bind glucocorticoids (GCs), is transcriptionally inactive, and is a potential inhibitor of activated GRalpha. Differential expression of GR isoforms may play a role in generalized or tissue-specific GC resistance. GCs induce apoptosis in neoplastic lymphoid cells; and, defective apoptosis is implicated in the genesis of chronic lymphocytic leukemia (CLL). We studied a patient with generalized GC resistance and CLL. GR number in the patient's transformed lymphocytes was approximately one half that of control cells with a approximately 10-fold reduction in binding affinity for dexamethasone. In vitro apoptosis induction in CLL cells was delayed in response to GCs, but not to other apoptosis inducers. Sequencing of the GR cDNA and gene including the 2.3-kb coding region, the intron/exon junctions, the known 5'-regulatory region, and approximately 300 bp of the 3'-region revealed no alterations. Western blot with an N-terminal antibody showed normal levels of immunoreactive GR, but quantitative analysis with isoform-specific C-terminal antibodies revealed a markedly reduced GRalpha expression, and high GRbeta expression. These findings indicate that imbalanced expression of the GR isoforms may be a mechanism of GC resistance, and may have implications for tumorigenesis by enhancing cell survival.

IFN-gamma induces cell growth inhibition by Fas-mediated apoptosis: requirement of STAT1 protein for up-regulation of Fas and FasL expression.

The mechanism by which IFN-gamma inhibits tumor cell growth has not been fully understood. Here we report that IFN-gamma up-regulated the expression of Fas and Fas ligand (FasL) on HT29 cells, a human colon adenocarcinoma cell line, and subsequently induced apoptosis of these cells. The kinetics of cell death in IFN-gamma-treated HT29 cells paralleled the increase in the levels of Fas and FasL expression. We further show that IFN-gamma up-regulated the expression of Fas and FasL in STAT1-transfected U3A cells but not in STAT1-deficient U3A cells. Correspondingly, IFN-gamma induced cell death in STAT1-transfected U3A cells but not in STAT1-deficient U3A cells. IFN-gamma-induced cell death was inhibited by caspase-1 inhibitors. Our results suggest that cell growth inhibition by IFN-gamma is due to apoptosis mediated by Fas and FasL interaction.

Modulation of tumoricidal function in alveolar macrophages from lung cancer patients by interleukin-6.

Previous studies have demonstrated that alveolar macrophages from lung cancer patients are impaired in their ability to develop tumoricidal function when stimulated by activators such as interferon gamma + lipopolysaccharide. However, these same macrophages have been shown to develop significant tumoricidal function when precultured with macrophage-depleted allogeneic peripheral blood lymphocytes from normal donors, an effect that was lost by the elimination of natural killer cells from the allogeneic lymphocyte population. In the present study, the effect of each activation condition on the expression of mRNA for interleukin-1 alpha (IL-1 alpha), IL-1 beta, tumor necrosis factor alpha (TNF alpha) and IL-6 was determined using reverse transcription/polymerase chain reaction. The results show that the non-permissive activation condition is associated with the expression of mRNA for IL-6 while the permissive activation condition is not. Antibodies against IL-6 were subsequently shown to permit the development of tumoricidal function in alveolar macrophages stimulated with interferon gamma + lipopolysaccharide while IL-6 protein was shown to inhibit the stimulatory action of allogeneic lymphocytes on the development of tumoricidal function in the same alveolar macrophages. Neither the permissive (i.e. allogeneic lymphocyte stimulation) nor the non-permissive (i.e. interferon gamma + lipopolysaccharide) activation condition had any effect on the capacity of alveolar macrophages from lung cancer patients to express mRNA for IL-1 alpha, IL-1 beta or TNF alpha. These results show that IL-6 can regulate the ability of alveolar macrophages from lung cancer patients to be stimulated by interferon gamma + lipopolysaccharide to develop significant tumoricidal function. They also show that allogeneic lymphocytes have the capacity to down-regulate IL-6 mRNA synthesis by alveolar macrophages thereby permitting the development and/or expression of macrophage tumoricidal function.

Selective inhibitors of 5-lipoxygenase reduce CML blast cell proliferation and induce limited differentiation and apoptosis.

Inhibitors of the arachidonic acid metabolizing enzyme, 5-lipoxygenase, reduce the rate of proliferation of chronic myelogenous leukemia blast cells. The inhibitory agents studied were ETYA, A63162 and SC41661A. These reagents induced differentiation of cultured chronic myelogenous leukemia cells from blast to promyelocytic morphology. Promyelocytic cells then underwent apoptosis, which was identified by nuclear and cytoplasmic morphological features and by DNA laddering. Proliferation of monoblastoid U937 and myelomonocytic HL60 cell lines, known to contain 5-lipoxygenase and synthesize leukotrienes, was reduced by these inhibitors. U937 cells cultured with ETYA, A63162 or SC41661A for 48 h exhibited apoptosis as assessed by DNA laddering and morphology. Characteristic ultrastructural changes of apoptosis were seen at 120 h. MK886, an inhibitor of 5-lipoxygenase with a mechanism of action distinct from oxidation/reduction reagents, at 20-40 microM also inhibited CML and U937 cell proliferation and induced apoptosis, as shown by DNA laddering and ultrastructure.

Apoptosis and interleukin 7 gene expression in chronic B-lymphocytic leukemia cells.

mRNA for interleukin 7 (IL-7) was readily detected in leukemic cells immediately upon their removal from patients with chronic B-lymphocytic leukemia (B-CLL). IL-7 mRNA expression and IL-7 gene transcription were down regulated, however, when B-CLL cells were placed in culture at 37 degrees C for 4 hr. Down regulation of the IL-7 gene was prevented in cells maintained at 4 degrees C. Continued culture of B-CLL cells at 37 degrees C resulted in programmed cell death, or apoptosis, as evidenced by DNA fragmentation. The coincident kinetics of IL-7 gene down regulation and apoptosis suggested that IL-7 gene expression may be required for maintenance of CLL viability in vivo. Signals for IL-7 gene regulation and apoptosis induction were thus examined. Activation of normal B cells through their immunoglobulin receptors did not result in upregulation of IL-7 gene expression. Reagents required for CLL cell purification and culture also did not contribute to IL-7 gene regulation and apoptosis induction. IL-7 gene expression was retained and apoptosis was prevented, however, in CLL cells cultured on a monolayer of EA.hy926 human umbilical cord endothelial hybrid cells. Signals specifically presented by EA.hy926 cells supported both CLL cell viability and IL-7 gene expression, whereas culture of CLL cells on A549/8 carcinoma cells, the fusion partner used to generate the EA.hy926 cells, did not. Cell-cell contact was required, as culture supernatants did not prevent apoptosis. Specifically, IL-7 mRNA expression was retained and apoptosis was prevented only by contact with the endothelial cell hybrids. Preliminary data indicated that integrins expressed on CLL cells affected modulation of apoptosis and IL-7 gene regulation, suggesting that integrins may play significant roles in regulating viability of CLL cells.