Mouth breathing, another risk factor for asthma: the Nagahama Study.

BACKGROUND: Allergic rhinitis, a known risk factor for asthma onset, often accompanies mouth breathing. Mouth breathing may bypass the protective function of the nose and is anecdotally considered to increase asthma morbidity. However, there is no epidemiological evidence that mouth breathing is independently associated with asthma morbidity and sensitization to allergens. In this study, we aimed to clarify the association between mouth breathing and asthma morbidity and allergic/eosinophilic inflammation, while considering the effect of allergic rhinitis. METHODS: This community-based cohort study, the Nagahama Study, contained a self-reporting questionnaire on mouth breathing and medical history, blood tests, and pulmonary function testing. We enrolled 9804 general citizens of Nagahama City in the Shiga Prefecture, Japan. RESULTS: Mouth breathing was reported by 17% of the population and was independently associated with asthma morbidity. The odds ratio for asthma morbidity was 1.85 (95% CI, 1.27-2.62) and 2.20 (95% CI, 1.72-2.80) in subjects with mouth breathing alone and allergic rhinitis alone, which additively increased to 4.09 (95% CI, 3.01-5.52) when mouth breathing and allergic rhinitis coexisted. Mouth breathing in nonasthmatics was a risk for house dust mite sensitization, higher blood eosinophil counts, and lower pulmonary function after adjusting for allergic rhinitis. CONCLUSION: Mouth breathing may increase asthma morbidity, potentially through increased sensitization to inhaled allergens, which highlights the risk of mouth-bypass breathing in the 'one airway, one disease' concept. The risk of mouth breathing should be well recognized in subjects with allergic rhinitis and in the general population.

Fucosylated TGF-ß receptors transduces a signal for epithelial-mesenchymal transition in colorectal cancer cells.

BACKGROUND: Transforming growth factor-ß (TGF-ß) is a major inducer of epithelial-mesenchymal transition (EMT) in different cell types. TGF-ß-mediated EMT is thought to contribute to tumour cell spread and metastasis. Sialyl Lewis antigens synthesised by fucosyltransferase (FUT) 3 and FUT6 are highly expressed in patients with metastatic colorectal cancer (CRC) and are utilised as tumour markers for cancer detection and evaluation of treatment efficacy. However, the role of FUT3 and FUT6 in augmenting the malignant potential of CRC induced by TGF-ß is unclear. METHODS: Colorectal cancer cell lines were transfected with siRNAs for FUT3/6 and were examined by cell proliferation, invasion and migration assays. The expression and phosphorylation status of TGF-ß downstream molecules were analysed by western blot. Fucosylation of TGF-ß receptor (TßR) was examined by lectin blot analysis. RESULTS: Inhibition of FUT3/6 expression by siRNAs suppressed the fucosylation of type I TßR and phosphorylation of the downstream molecules, thereby inhibiting the invasion and migration of CRC cells by EMT. CONCLUSION: Fucosyltransferase 3/6 has an essential role in cancer cell adhesion to endothelial cells by upregulation of sialyl Lewis antigens and also by enhancement of cancer cell migration through TGF-ß-mediated EMT.

Three-dimensional patterns of early acetabular cartilage damage in hip dysplasia; a high-resolutional CT arthrography study.

OBJECTIVE: The purpose of this study was to examine the three-dimensional (3D) progression patterns of early acetabular cartilage damage in hip dysplasia using high-resolutional computed tomography (CT) arthrography. DESIGN: Thirty-two dysplastic hips of 26 Japanese symptomatic females including 21 hips in pre-stage of osteoarthritis (Kellgren-Lawrence (K-L) grade 0; mean patient age, 32.0 years) and 11 hips in early stage of osteoarthritis (K-L grade 1 or 2; mean patient age, 32.8 years) were examined. Isotropic high-resolutional CT arthrography with an image resolution of 0.5 mm in any orthogonal direction was performed. A 3D acetabular cartilage model was generated and we evaluated distribution of cartilage thickness in 12 zones after dividing the weight-bearing area of the hip joint in radial and lateral/medial directions. RESULTS: In pre-stage of osteoarthritis, significant differences in cartilage thickness were observed between the lateral and medial zones in all radial regions, most prominently in the antero-superior region. In early stage of osteoarthritis, no significant differences in cartilage thickness were observed, except in the most posterior region. The lateral-medial (LM) ratio was defined as cartilage thickness in the lateral zone divided by that in the medial zone, and hips with the LM ratio in the antero-superior region of <1.4 had significantly more extensive involvement of labral tears than hips with the LM ratio of ≥1.4. CONCLUSIONS: In hip dysplasia, acetabular cartilage damage was probably occurred in the antero-superior lateral area. The LM ratio may be a sensitive index to quantify early cartilage damage associated with extent of labral disorders.

Influence of medial meniscectomy on stress distribution of the femoral cartilage in porcine knees: a 3D reconstructed T2 mapping study.

OBJECTIVE: Previous studies have shown that meniscectomy results in an increase of local load transmission and may cause degeneration of the knee cartilage. Using 3D reconstructed T2 mapping, we examined the influence on the femoral cartilage under loading after medial meniscectomy. DESIGN: Ten porcine knees were imaged using a pressure device and a 3.0-T magnetic resonance imaging (MRI) system. Consecutive sagittal T2 maps were obtained in neutral alignment with and without compression, and under compression at 10° varus alignment. After medial meniscectomy, the aforementioned MRI was repeated. Cartilage T2 before and after meniscectomy under each condition were compared at the 12 regions of interest (ROIs) defined on the 3D weight-bearing area of the femoral cartilage. RESULTS: Before meniscectomy, large decreases in T2 under neutral compression were mainly seen at the anterior and central ROIs of the medial cartilage, which shifted to the posterior ROIs after meniscectomy. There were significant differences in decrease in T2 ratio with loading before and after meniscectomy (9.8%/4.3% at the anterior zone, 4.0%/11.4% at the posterior zone, P < 0.05). By applying varus compression, a more remarkable decrease in the cartilage T2 in posterior ROIs after meniscectomy was achieved. (Before/after meniscectomy: 8.7%/2.5% at the anterior zone, 7.2%/18.7% at the posterior zone, P < 0.05). CONCLUSIONS: Assuming a decrease in T2 with loading correlated with the applied pressure, a deficiency of the medial meniscus resulted in a shift of the primary area with a maximal decrease of cartilage T2 with loading posteriorly in the porcine knee joint, presumably reflecting the intraarticular environment of load transmission.

Typing of O26 enterohaemorrhagic and enteropathogenic Escherichia coli isolated from humans and cattle with IS621 multiplex PCR-based fingerprinting.

AIMS: This study evaluated a typing method of O26:H11 enterohaemorrhagic and enteropathogenic Escherichia coli (EHEC and EPEC) based on the variation in genomic location and copy numbers of IS621. METHODS AND RESULTS: Two multiplex PCRs, targeting either the left (5') or right (3') IS/chromosome junction of 12 IS621 insertion sites and one PCR specific of another truncated copy, were developed. Thirty-eight amplification profiles were observed amongst a collection of 69 human and bovine O26:H11 EHEC and EPEC. Seventy-one per cent of the 45 EHEC and EPEC with identical IS621 fingerprints within groups of two, three or four isolates had >85% pulsed field gel electrophoresis (PFGE) profile similarity, including four groups of epidemiologically related EHEC or EPEC, while most of the groups had <85% similarity between each others. Epidemiologically related EHEC from each of three independent outbreaks in Japan and Belgium also exhibited identical IS621 fingerprints and PFGE profiles. CONCLUSIONS: The IS621 fingerprinting and the PFGE are complementary typing assays of EHEC and EPEC; though, the former is less discriminatory. SIGNIFICANCE AND IMPACT OF THE STUDY: The IS621 printing method represents a rapid (24 h) first-line surveillance and typing assay, to compare and trace back O26:H11 EHEC and EPEC during surveys in farms, multiple human cases and outbreaks.

Loading and knee alignment have significant influence on cartilage MRI T2 in porcine knee joints.

OBJECTIVE: Physiological magnetic resonance imaging (MRI) under loading or knee malalignment conditions has not been thoroughly investigated. We assessed the influence of static loading and knee alignment on T2 (transverse relaxation time) mapping of the knee femoral cartilage of porcine knee joints using a non-metallic pressure device. METHODS: Ten porcine knee joints were harvested en bloc with intact capsules and surrounding muscles and imaged using a custom-made pressure device and 3.0-T MRI system. Sagittal T2 maps were obtained (1) at knee neutral alignment without external loading (no loading), (2) under mechanical compression of 140 N (neutral loading), and (3) under the same loading conditions as in (2) with the knee at 10 degrees varus alignment (varus loading). T2 values of deep, intermediate, and superficial zones of the medial and lateral femoral cartilages at the weight-bearing area were compared among these conditions using custom-made software. Cartilage contact pressure between the femoral and tibial cartilages, measured by a pressure-sensitive film, was correlated with cartilage T2 measurements. RESULTS: In the medial cartilage, mean T2 values of the deep, intermediate, and superficial zones decreased by 1.4%, 13.0%, and 6.0% under neutral loading. They further decreased by 4.3%, 19.3%, and 17.2% under varus loading compared to no loading. In the lateral cartilage, these mean T2 values decreased by 3.9%, 7.7%, and 4.2% under neutral loading, but increased by 1.6%, 9.6%, and 7.2% under varus loading. There was a significant decrease in T2 values in the intermediate zone of the medial cartilage under both neutral and varus loading, and in the superficial zone of the medial cartilage under varus loading (P<0.05). Total contact pressure values under neutral loading and varus loading conditions significantly correlated with T2 values in the superficial and intermediate zones of the medial cartilages. CONCLUSIONS: The response of T2 to change in static loading or alignment varied between the medial and lateral cartilages, and among the deep, intermediate, and superficial zones. These T2 changes were significantly related to the contact pressure measurements. Our results indicate that T2 mapping under loading allows non-invasive, biomechanical assessment of site-specific stress distribution in the cartilage.

Horizontal propagation of excitation in rat visual cortical slices revealed by optical imaging.

Optical imaging with high spatial and temporal resolution of neural activity in rat cortical slices was used to investigate the dynamics of signal transmission through neural connections in the visual cortex. When inhibition due to gamma-aminobutyric acid was slightly suppressed, horizontal propagation of excitation in both the supra- and infragranular layers became prominent. This propagation was not affected by vertical cuts in either the supra- or infragranular layer, which suggests that excitation is at least partially conveyed horizontally by reciprocal vertical connections between neurons in these layers.

Promotion of medial collateral ligament healing using a high-voltage electrical field.

PURPOSE: To assess the biomechanical and histological effects of a high-voltage electrical field on ligament healing. METHODS: 54 male Wistar rats were used. The medial collateral ligaments (MCLs) of 6 rats were measured for normal biomechanical failure load and stiffness. MCLs of the remaining 48 rats were transected and randomly assigned; half were subjected to a high-voltage electrical field for 8 hours a day (experimental group), and the other half were kept under normal conditions (control group). Eight rats from each group were killed 2, 4, and 6 weeks after the transection and their MCLs were tested biomechanically and histologically. RESULTS: Failure load and stiffness of the MCLs in the experimental group were significantly higher than those in the control group 2 weeks after transection, but not thereafter. At week 2, the granulation tissue completely occupied the space between the cut ends of the MCL in the experimental group, whereas some gaps remained in the control group. CONCLUSION: A high-voltage electrical field appear to accelerate ligament healing in the initial 2 weeks. Earlier initial healing may allow earlier start of the rehabilitation.

Composite hypo-hyper-d-intermetallic and interionic phases as supported interactive electrocatalysts.

Interactive, strong interbonding and highly electron conductive nonstoichiometric titanium suboxide catalytic supports, Magneli phases (Ti(n)O(2n-1), on average Ti(4)O(7)), have been used in the electrocatalysis of hydrogen (HELR) and oxygen (OELR) electrode reactions with remarkable consequences and advanced achievements. The theory of hypo-hyper-d-interelectronic bonding of transition metal ions and atoms has been employed for selective ordered grafting and shown to stay in the core of the strong metal-support interaction (SMSI) in heterogeneous catalysis and electrocatalysis, and thereby the substantial cause for the improved synergistic activity of composite (electro)catalysts. The same fundament has been the thermodynamic basis for the thermal production of symmetric intermetallic Laves type phases of nanostructured electrocatalysts, in particular the ones with higher oxophilic properties of hypo-d-elements. Remarkably advanced in electrocatalytic activity, highly monatomically dispersed deposits of Pt upon Magneli phases are shown to be unique and highly promising electrocatalysts for the cathodic oxygen reduction (ORR). Nanostructured Au upon a thin nanocrystalline film of anatase titania has been confirmed by X-ray photoelectron spectroscopy (XPS) as a typical classical paradigm of the SMSI, and at the same time affording the basis for gold with strained d-orbitals, as the reversible hydrogen electrode. Magneli phases have been shown to be the best electrocatalytic supports with unique properties both for low temperature PEM fuel cells (LT PEM FCs) with pronounced CO tolerance and water electrolysis in membrane type hydrogen generators.

Relationship between variations in the circle of Willis and flow rates in internal carotid and basilar arteries determined by means of magnetic resonance imaging with semiautomated lumen segmentation: reference data from 125 healthy volunteers.

BACKGROUND AND PURPOSE: Volume flow rates in the feeding arteries of the brain are measured to evaluate blood flow dynamics in vascular disease. Although these flow values are thought to be effected by anatomic variations in the circle of Willis, few reports have described the effect. This study reports on the relationship between variations in the circle of Willis and volume flow rates in the bilateral internal carotid and basilar arteries of normal volunteers. METHODS: We prospectively examined 125 healthy volunteers by MR imaging. Variations in the circle of Willis were classified as "textbook" type, hypoplasia of the precommunicating segment of the anterior cerebral artery (A1), hypoplasia of the precommunicating segment of the posterior cerebral artery (P1), or "other." Volume flow rates were measured by 2D cine phase-contrast MR imaging. Lumen boundaries and volume flow rates were semiautomatically determined by pulsatility-based segmentation. RESULTS: Of the 117 subjects (61 men, 56 women; mean age, 23.6 years) considered suitable for flow measurement, 105 showed textbook type, and 6 each showed A1 hypoplasia and P1 hypoplasia. Total flow rates for the 3 variations were 781 +/- 151 mL/min (mean +/- SD), 744 +/- 119, and 763 +/- 129, respectively. Relative contributions by flow rates of the internal carotid arteries and the basilar artery for the 3 variations were 39.8%:38.9%:21.3%, 31.8%:49.1%:19.0%, and 46.6%:41.6%:11.7%, respectively, showing statistically significant differences. CONCLUSIONS: Variations in the circle of Willis correlate significantly with relative contributions by the flow rates of the bilateral internal carotid and basilar arteries.

Comparison of patient doses in 256-slice CT and 16-slice CT scanners.

The 256-slice CT-scanner has been developed at the National Institute of Radiological Sciences. Nominal beam width was 128 mm in the longitudinal direction. When scanning continuously at the same position to obtain four-dimensional (4D) images, the effective dose is increased in proportion to the scan time. Our purpose in this work was to measure the dose for the 256-slice CT, to compare it with that of the 16-slice CT-scanner, and to make a preliminary assessment of dose for dynamic 3D imaging (volumetric cine imaging). Our group reported previously that the phantom length and integration range for dosimetry needed to be at least 300 mm to represent more than 90% of the line integral dose with the beam width between 20 mm and 138 mm. In order to obtain good estimates of the dose, we measured the line-integral dose over a 300 mm range in PMMA (polymethylmethacrylate) phantoms of 160 mm or 320 mm diameter and 300 mm length. Doses for both CT systems were compared for a clinical protocol. The results showed that the 256-slice CT generates a smaller dose than the 16-slice CT in all examinations. For volumetric cine imaging, we found an acceptable scan time would be 6 s to 11 s, depending on examinations, if dose must be limited to the same values as routine examinations with a conventional multidetector CT. Finally, we discussed the studies necessary to make full use of volumetric cine imaging.

Effectiveness of flexible noise control image processing for digital portal images using computed radiography.

The purpose of this study was to evaluate the clinical efficacy of flexible noise control (FNC) image processing in off-line computed radiography (CR) portal images. An observer study was designed to compare FNC with multiobjective frequency processing (MFP) in images acquired using a CR portal imaging device (CRPID). The image processing of MFP and FNC used the same data and required no additional irradiation of patients, and all images were printed on 10-bit grey-level dry laser film. Four radiation technologists and one oncologist served as observers and evaluated 40 sets of images for three different treatment sites: brain, lung and pelvis. Six to 10 anatomical landmarks were selected from each treatment site. Each observer was asked to rate each landmark in terms of its clinical visibility and rate the ease of making the pertinent verification in the MFP- and FNC-processed images. In the ratings of the visibility of landmarks and for the verification of treatment ports, FNC-processed images were more visible than MFP-processed images except for several landmarks in the anteroposterior (AP) pelvis such as the pubic symphysis. The visibility of landmarks in FNC-processed images was comparable with that in MFP-processed images. The verification of treatment ports using the CRPID with FNC was generally achievable. In conclusion, this study suggests that FNC is effective for image processing of CR portal images.

Chaotic dynamics of a behavior-based miniature mobile robot: effects of environment and control structure.

To study the regularity and complexity of autonomous behavior, the flow of sensory information obtained in autonomous mobile robots under various conditions was analyzed as a complex system. Sensory information time series Xn was collected from a miniature mobile robot during free navigation, and plotted on the return map, the graph of X(n+tau) vs. Xn. The plot exhibited a characteristic trajectory, representing the regularity of the time series. Correlation integral and Lyapunov exponent analysis also showed properties of deterministic chaos; the presence of fractal dimension and positive Lyapunov exponent. Analysis of sensory information obtained in the robot with three different neural controllers revealed that the autonomous robot behaves in such a way that the flow of sensory information is governed by a deterministic rule, and this pattern is unique to each controller. Furthermore, the analysis in various environments exhibited that transitions from one trajectory to another on the return map occur during the course of autonomous behavior. The fractal and Lyapunov dimensions calculated in various conditions indicate that these dimension could be utilized to quantify the complexity of autonomous behavior and the relative difficulty of tasks. Analyses at different evolutionary stage revealed that behavioral performance correlates with fractal dimension. These studies using a miniature mobile robot that allowed to idealize the experimental conditions demonstrated firmly that the complex analysis could be utilized in evaluation and optimization of autonomous systems and the behavior.

Stimulation of nerve growth factor synthesis/secretion by 1,4-benzoquinone and its derivatives in cultured mouse astroglial cells.

Previously we reported that astroglial cells cultured from mouse brain synthesize and secrete nerve growth factor (NGF) and that, in quiescent cells, catecholamines markedly increase the NGF content in the conditioned medium (CM). We wished to further assess the structural properties required for exhibition of such effect of compounds containing a ring structure analogous to that of catechol on astroglial NGF synthesis. During our study, we found that hydroquinone, which was confirmed not to stimulate NGF synthesis in mouse fibroblast cells in another of our investigations, is a potent stimulator of NGF synthesis in astroglial cells and that 1,4-benzoquinone, an oxidized form of hydroquinone, is a more effective stimulator than hydroquinone itself. In addition, the results of experiments with 1,2-benzoquinone derivatives indicated that the presence of a long aliphatic side chain in the molecule eliminates the stimulatory effect of 1,4-benzoquinone on NGF synthesis in astroglial cells.

Tumor necrosis factor is markedly synergistic with interleukin 1 and interferon-gamma in stimulating the production of nerve growth factor in fibroblasts.

A possible interaction between tumor necrosis factor-alpha (TNF) and other cytokines/growth factors in stimulating the production of nerve growth factor (NGF) in Swiss 3T3 cells was studied. TNF's stimulatory activity on fibroblast NGF production was synergized by interleukin-1 alpha (IL-1 alpha), IL-1 beta and interferon-gamma (IFN-gamma), but was antagonized by transforming growth factor-beta (TGF-beta). The most remarkable synergistic effect was observed between TNF and IL-1 alpha/beta; as little as 0.003 ng/ml of IL-1 beta markedly enhanced TNF's stimulatory activity on NGF production in the cells. These findings reinforce the idea that TNF, in concert with IL-1 alpha/beta, plays an essential role in regulating the regeneration of peripheral nerves following injury through an indirect mechanism by which it stimulates NGF production in fibroblasts.

Effect of soymilk consumption on serum estrogen and androgen concentrations in Japanese men.

Soy consumption has been associated with a reduced risk of prostate cancer. The mechanism for this association may involve the effect of soy on the endocrine system. We conducted a randomized dietary intervention study to determine the effects of soy consumption on serum levels of steroid hormones in men. Thirty-five men were randomly assigned to either a soymilk-supplemented group or a control group. The men in the soy-supplemented group were asked to consume 400 ml of soymilk daily for 8 weeks. The men in the control group maintained their usual diet. Blood samples were obtained just before the initiation of the dietary period and thereafter every two weeks for 12 weeks. Changes in hormone concentrations were analyzed and compared between the two groups using the mixed linear regression model against weeks from the start of the dietary period. The mean (SD) soymilk intake estimated from dietary records during the dietary study period was 342.9 (SD, 74.2) ml in the soymilk-supplemented group. There was a significant difference between the two groups in terms of changes in serum estrone concentrations, which tended to decrease in the soy-supplemented group and increase in the control group over time. None of the other hormones measured (estradiol, total and free-testosterone, or sex hormone-binding globulin) showed any statistical difference between the two groups in terms of patterns of change. The results of the study indicate that soymilk consumption may modify circulating estrone concentrations in men.

Imidazo[2,1-b]benzothiazoles. 2. New immunosuppressive agents.

A series of 2-phenylimidazo[2,1-b]benzothiazole derivatives was prepared and tested for immunological activities. Some of the compounds showed significant suppressive activity of delayed type hypersensitivity (DTH) without inhibition of humoral immunity in mice by oral administration. The most active compound was 2-(m-hydroxyphenyl)imidazo[2,1-b]benzothiazole (20).

6-(1H-imidazol-1-yl)-7-nitro-2,3(1H,4H)-quinoxalinedione hydrochloride (YM90K) and related compounds: structure-activity relationships for the AMPA-type non-NMDA receptor.

A novel series of quinoxalinediones possessing imidazolyl and related heteroaromatic substituents was synthesized and evaluated for their activity to inhibit [3H]AMPA binding from rat whole brain. From the structure-activity relationships, it was found that the 1H-imidazol-1-yl moiety could function as a bioisostere for the cyano and nitro groups, and that 6-(1H-imidazol-1-yl)-7-nitro-2,3(1H,4H)-quinoxalinedione (11) showed the most potent activity for the AMPA receptor. Compound 11 was evaluated for selectivity versus other excitatory amino acid receptors, and its action against AMPA at its receptor in the rat striatum was characterized. These data showed that compound 11 was a selective antagonist for the AMPA receptor with a Ki value of 0.084 microM, being approximately equipotent with 2,3-dihydro-6-nitro-7-sulfamoylbenzo(f)quinoxaline (3) (NBQX; Ki = 0.060 microM). Compound 11 was also found to give protection against sound-induced seizure on DBA/2 mice at the minimum effective dose of 3 mg/kg ip (3; 10 mg/kg ip).

Alterations in [3H]MK-801, [3H]muscimol, [3H]cyclic AMP, and [3H]rolipram binding in the gerbil hippocampus following repeated ischemic insults.

Using [3H]MK-801, [3H]muscimol, [3H]cyclic AMP, and [3H]rolipram, we performed quantitative in vitro autoradiography to determine sequential alterations in the binding of N-methyl-D-aspartate and GABAA receptors, particulate cyclic AMP-dependent protein kinase, and cyclic AMP-selective phosphodiesterase, respectively, in the gerbil hippocampus following repeated brief ischemic insults. Changes from 1 h to 28 days after three 2-min ischemic insults at 1-h intervals were compared with those after 2 and 6 min of ischemia. We observed no alterations in the binding of all the four ligands throughout the observation period following 2 min of ischemia which produced no histological neuronal damage except for transient reductions in [3H]cyclic AMP binding during the early reperfusion period. [3H]Cyclic AMP binding in the CA1 subfield decreased one day after 6 min of ischemia and four days after three 2-min ischemic insults, and 62-65% of the binding was lost after 28 days. [3H]Rolipram binding in the CA1 subfield decreased one day after 6 min of ischemia and the binding was reduced by 45-50% after four and 28 days. Following three 2-min ischemic insults, [3H]rolipram binding decreased in the CA1 at one day, and decreased by 25-33% after 28 days. Both [3H]MK-801 and [3H]muscimol binding was preserved during the early reperfusion period after 6 min of ischemia and three 2-min ischemic insults. Reductions in [3H]MK-801 binding in CA1 were observed four days after ischemic insults when CA1 neuronal destruction was seen. After one month, approximately 50% of [3H]MK-801 binding was lost in CA1 in both groups.(ABSTRACT TRUNCATED AT 250 WORDS)