Integration of the Duke Activity Status Index into preoperative risk evaluation: a multicentre prospective cohort study.

BACKGROUND: The Duke Activity Status Index (DASI) questionnaire might help incorporate self-reported functional capacity into preoperative risk assessment. Nonetheless, prognostically important thresholds in DASI scores remain unclear. We conducted a nested cohort analysis of the Measurement of Exercise Tolerance before Surgery (METS) study to characterise the association of preoperative DASI scores with postoperative death or complications. METHODS: The analysis included 1546 participants (≥40 yr of age) at an elevated cardiac risk who had inpatient noncardiac surgery. The primary outcome was 30-day death or myocardial injury. The secondary outcomes were 30-day death or myocardial infarction, in-hospital moderate-to-severe complications, and 1 yr death or new disability. Multivariable logistic regression modelling was used to characterise the adjusted association of preoperative DASI scores with outcomes. RESULTS: The DASI score had non-linear associations with outcomes. Self-reported functional capacity better than a DASI score of 34 was associated with reduced odds of 30-day death or myocardial injury (odds ratio: 0.97 per 1 point increase above 34; 95% confidence interval [CI]: 0.96-0.99) and 1 yr death or new disability (odds ratio: 0.96 per 1 point increase above 34; 95% CI: 0.92-0.99). Self-reported functional capacity worse than a DASI score of 34 was associated with increased odds of 30-day death or myocardial infarction (odds ratio: 1.05 per 1 point decrease below 34; 95% CI: 1.00-1.09), and moderate-to-severe complications (odds ratio: 1.03 per 1 point decrease below 34; 95% CI: 1.01-1.05). CONCLUSIONS: A DASI score of 34 represents a threshold for identifying patients at risk for myocardial injury, myocardial infarction, moderate-to-severe complications, and new disability.

Medical education from the point of view of medical students: Results from four participatory Delphi panels in Quito, Ecuador.

BACKGROUND: Medical curricula have historically been designed in a top-down approach, usually excluding students. While Delphi panels have been used as a tool for medical education curricula design, none have been conducted in Ecuador. In addition, no such approach has ever included students both as panelists and researchers. MATERIAL AND METHODS: Four Delphi panels were developed and conducted using a participatory approach that allowed medical students to take part both as expert panelists and researchers: specifically, students developed the questionnaire and conducted a qualitative synthesis. Questionnaire responses were anonymized and dispatched online to panelists. The information was organized and collected to develop the qualitative syntheses and prepare the final statements. RESULTS: Thirty-two medical students participated between February and May 2018. A total of 32 questions were developed, corresponding to five different categories. For some questions, consensus was reached; for other questions, general statements were obtained.Discussion and conclusion: Developing the questionnaire, responding to it and analyzing the answers allowed students to raise significant concerns regarding medical education topics proposing relevant policy and curricula change. Participatory Delphi panels can be an efficient tool to obtain organized feedback, improve student class involvement, and promote research skills.

Altitude effects on technology and productivity of small bovine farms (milk meat) in Veracruz (Gulf of Mexico).

The dual-purpose bovine system represents 98.4% of the bovine livestock of Veracruz, the main cattle-producing state of Mexico. This system supplies calves to meat companies, a sector in which Veracruz has been the national leader in the last decade. The objective of the present study was to analyze the effect of the altitudinal zonation of farms on livestock technology and productivity in a microbasin of the Gulf of Mexico where small farms predominate. Structured interviews were applied to producers located in three altitudinal zones (at average altitudes of 50, 140, and 450 m, respectively, for lower, middle, and upper zones). Sample size was 135 farms having similar land surface (within a range of 15-22 ha). The results indicated multiple differences among farms located in the three zones. Farms in the middle and lower zones presented higher productive indicators than those in the upper zone. Differences in herd structure and management resulted in important differences in productivity, income, and profits in milk and calf production. We concluded from this study that altitudinal zonation in Veracruz had a clear effect on the differentiation of small farms, which are representative of dual-purpose cattle. The upper zone performs cattle activity under conditions with greater disadvantages in the analyzed region.

Reductions of plant cover induced by sheep grazing change the above-belowground partition and chemistry of organic C stocks in arid rangelands of Patagonian Monte, Argentina.

The objective of this study was to estimate the size and chemical quality of the total organic C stock and its partition between above-belowground plant parts and soil at sites with different plant cover induced by sheep grazing in the arid Patagonian Monte. This study was conducted at six representative sites with increasing signs of canopy disturbance attributed to grazing pressure. We used faeces density as a proxy of grazing pressure at each site. We assessed the total plant cover, shrub and perennial grass cover, total standing aboveground biomass (AGB), litter mass and belowground biomass (BGB) at each site. We further estimated the content of organic C, lignin and soluble phenols in plant compartments and the content of organic C, organic C in humic substances (recalcitrant C) and water soluble C (labile C) in soil at each site. Total plant cover was significantly related to grazing pressure. Standing AGB and litter mass decreased with increasing canopy disturbance while BGB did not vary across sites. Total organic C stock and the organic C stock in standing AGB increased with increasing total plant, shrub, and perennial grass cover. The organic C stock in litter mass increased with increasing total plant and shrub cover, while the organic C stock in BGB did not vary across sites. Lignin content in plant compartments increased with increasing total and shrub cover, while soluble phenols content did not change across sites. The organic C stock and the water soluble C content in soil were positively associated with perennial grass cover. Changes in total plant cover induced by grazing pressure negatively affected the size of the total organic C stock, having minor impact on the size of belowground than aboveground components. The reduction of perennial grass cover was reflected in decreasing chemical quality of the organic C stock in soil. Accordingly, plant managerial strategies should not only be focused on the amount of organic C sequestered but also on the chemical quality of organic C stocks since C chemistry could have an important impact on ecosystem functioning.

Meaningful traits for grouping plant species across arid ecosystems.

Grouping species may provide some degree of simplification to understand the ecological function of plants on key ecosystem processes. We asked whether groups of plant species based on morpho-chemical traits associated with plant persistence and stress/disturbance resistance reflect dominant plant growth forms in arid ecosystems. We selected twelve sites across an aridity gradient in northern Patagonia. At each site, we identified modal size plants of each dominant species and assessed specific leaf area (SLA), plant height, seed mass, N and soluble phenol concentration in green and senesced leaves at each plant. Plant species were grouped according with plant growth forms (perennial grasses, evergreen shrubs and deciduous shrubs) and plant morphological and/or chemical traits using cluster analysis. We calculated mean values of each plant trait for each species group and plant growth form. Plant growth forms significantly differed among them in most of the morpho-chemical traits. Evergreen shrubs were tall plants with the highest seed mass and soluble phenols in leaves, deciduous shrubs were also tall plants with high SLA and the highest N in leaves, and perennial grasses were short plants with high SLA and low concentration of N and soluble phenols in leaves. Grouping species by the combination of morpho-chemical traits yielded 4 groups in which species from one growth form prevailed. These species groups differed in soluble phenol concentration in senesced leaves and plant height. These traits were highly correlated. We concluded that (1) plant height is a relevant synthetic variable, (2) growth forms adequately summarize ecological strategies of species in arid ecosystems, and (3) the inclusion of plant morphological and chemical traits related to defenses against environmental stresses and herbivory enhanced the potential of species grouping, particularly within shrubby growth forms.

Effects of long-term grazing disturbance on the belowground storage of organic carbon in the Patagonian Monte, Argentina.

The objective of this study was to analyze the effect of grazing disturbance on the amount and the spatial distribution (vertical and horizontal) of root biomass and soil organic carbon (SOC) in order to evaluate whether grazing alters the belowground storage of organic carbon (C) in arid rangelands of the Patagonian Monte. We selected three representative sites (3 ha each) with low, moderate and high grazing disturbance located far, mid-distance and near the watering point, respectively, in rangelands submitted to sheep grazing for more than 100 years. We assessed the canopy structure and identified the four most frequent plant patch types at each site. We selected four replications of each patch type and extracted a soil sample (0-30 cm depth) underneath the canopy and in the middle of the nearest inter-patch bare soil area in winter and summer. We assessed the root and soil dry mass and the respective organic C concentration in each sample and then we estimated the total belowground organic C storage at each site. Total plant and perennial grass cover were lower with high than low grazing disturbance while the reverse occurred with dwarf shrub cover. High grazing disturbance led to the increase in total root biomass in the whole soil profile of patch areas and in the upper soil of inter-patch areas. SOC was higher in patch than in inter-patch areas at all sites but at both areas was reduced with high grazing disturbance. This was probably the result of the low total plant cover and the low and recalcitrant contribution of above and below-ground plant litter to soils at sites with high grazing disturbance. Accordingly, these changes did not result in variations in the total belowground organic C storage. We concluded that high grazing disturbance did not affect the total belowground organic C storage but led to changes in the spatial patterning of this organic C storage (i.e shifting from soil to roots).

Effectiveness of a specific follow up program for the management of the mental components of post-intensive care syndrome and chronic pain after COVID-19: results from the PAIN-COVID randomized clinical trial.

BACKGROUND: Critical COVID-19 survivors are at risk of developing Post-intensive Care Syndrome (PICS) and Chronic ICU-Related Pain (CIRP). We determined whether a specific care program improves the quality of life (QoL) of patients at risk of developing PICS and CIRP after COVID-19. METHODS: The PAIN-COVID trial was a parallel-group, single-centre, single-blinded, randomized controlled trial. The intervention consisted of a follow up program, patient education on PICS and pain, and a psychological intervention based on Rehm's self-control model in patients with abnormal depression scores (≥8) in the Hospital Anxiety and Depression Scale (HADS) at the baseline visit. QoL was evaluated with the 5-level EQ 5D (EQ 5D 5 L), mood disorders with the HADS, post-traumatic stress disorder (PTSD) with the PCL-5 checklist, and pain with the Brief Pain Inventory short form, the Douleur Neuropathique 4 questionnaire, and the Pain Catastrophizing Scale. The primary outcome was to determine if the program was superior to standard-of-care on the EQ visual analogue scale (VAS) at 6 months after the baseline visit. The secondary outcomes were EQ VAS at 3 months, and EQ index, CIRP incidence and characteristics, and anxiety, depression, and PTSD at 3 and 6 months after baseline visits. CONCLUSIONS: This program was not superior to standard care in improving QoL in critical COVID-19 survivors as measured by the EQ VAS. However, our data can help establish better strategies for the study and management of PICS and CIRP in this population. TRIAL REGISTRATION: # NCT04394169, registered on 5/19/2020.

Negative screening colonoscopy after a positive guaiac faecal occult blood test: not a contraindication to continued screening.

AIM: In guaiac faecal occult blood test (gFOBT) screening at least 50% of positive individuals will have a colonoscopy negative for colorectal neoplasia. The question of continuing screening in this group has not been addressed. METHOD: Data on participants aged 50-69 years with a positive gFOBT result and a negative colonoscopy were followed through the biennial screening pilot conducted between 2000 and 2007 in Scotland. RESULTS: In the first screening round, 1527 colonoscopies were negative for neoplasia. 1300 were re-invited in the second round, 905 accepted, and 157 had a positive gFOBT result, giving a positivity rate of 17.4%. Colonoscopy revealed 20 subjects with adenoma and six with invasive cancer. In the third screening round 1031 were invited for a third time and 730 accepted: 55 had a positive gFOBT test, giving a positivity rate of 7.5%. In this group, six colonoscopies revealed adenomas but there were no cancers diagnosed. In the third screening round, 108 individuals had had two positive gFOBT results and two subsequent negative colonoscopies. Eighty-four were invited for a third gFOBT, 66 accepted and 19 (25.6%) had a positive result none of whom had an adenoma or carcinoma. CONCLUSION: These data indicate that a negative colonoscopy following a positive gFOBT is not a contraindication for further screening, although this is likely to have a low yield of neoplastic pathology after two negative colonoscopies.

Role of the PI3K regulatory subunit in the control of actin organization and cell migration.

Cell migration represents an important cellular response that utilizes cytoskeletal reorganization as its driving force. Here, we describe a new signaling cascade linking PDGF receptor stimulation to actin rearrangements and cell migration. We demonstrate that PDGF activates Cdc42 and its downstream effector N-WASP to mediate filopodia formation, actin stress fiber disassembly, and a reduction in focal adhesion complexes. Induction of the Cdc42 pathway is independent of phosphoinositide 3-kinase (PI3K) enzymatic activity, but it is dependent on the p85alpha regulatory subunit of PI3K. Finally, data are provided showing that activation of this pathway is required for PDGF-induced cell migration on collagen. These observations show the essential role of the PI3K regulatory subunit p85alpha in controlling PDGF receptor-induced cytoskeletal changes and cell migration, illustrating a novel signaling pathway that links receptor stimulation at the cell membrane with actin dynamics.

Intraneural injection during nerve stimulator-guided sciatic nerve block at the popliteal fossa.

BACKGROUND: Exact location of the needle tip during nerve stimulation-guided peripheral nerve blocks is unknown. Using high-frequency ultrasound imaging, we tested the hypothesis that intraneural injection is common with nerve stimulator-guided sciatic nerve (SN) block in popliteal fossa. METHODS: Forty-two patients scheduled for hallux valgus repair were studied. Sciatic block at the popliteal fossa was accomplished using nerve stimulation. When a motor response was elicited at <0.5 mA (2 Hz, 0.1 ms), 40 ml of local anaesthetic (LA) was injected. Using ultrasound (Titan, Sonosite, 5-10 MHz), the diameters and area of the SN were measured before and after the injection. The presence of nerve swelling and proximal or distal diffusion of LA were also assessed. Intraneural injection was defined as nerve area (NA) increase of > OR =15% and one or more additional ultrasonographic markers (nerve swelling, proximal-distal diffusion within epineural tissue). Clinical neurological evaluation was performed 1 week after the block. RESULTS: Post-injection NA increase > OR =15% was seen in 32 (76%) patients [0.54 (SD 0.19) cm(-2) vs 0.76 (0.24) cm(-2); P<0.05]. Nerve swelling with fascicular separation was observed in 37 (88%) patients; proximal and distal diffusion of LA were present in six (14%) and 14 (38%) patients, respectively. Intraneural injection criteria were met in 28 (66%) patients. Greater NA increase was present in patients with fast block onset [61 (45) vs 25 (33)%; (Dif 35% 95% CI 61-9%); P<0.05]. No patient developed neurological complications. CONCLUSIONS: Intraneural (subepineural) injection is a common occurrence after nerve stimulator-guided SN block at the popliteal fossa, yet it may not inevitably lead to neurological complications.

Biradicalic excited states of zwitterionic phenol-ammonia clusters.

Phenol-ammonia clusters with more than five ammonia molecules are proton transferred species in the ground state. In the present work, the excited states of these zwitterionic clusters have been studied experimentally with two-color pump probe methods on the nanosecond time scale and by ab initio electronic-structure calculations. The experiments reveal the existence of a long-lived excited electronic state with a lifetime in the 50-100 ns range, much longer than the excited state lifetime of bare phenol and small clusters of phenol with ammonia. The ab initio calculations indicate that this long-lived excited state corresponds to a biradicalic system, consisting of a phenoxy radical that is hydrogen bonded to a hydrogenated ammonia cluster. The biradical is formed from the locally excited state of the phenolate anion via an electron transfer process, which neutralizes the charge separation of the ground state zwitterion.

Constructed wetlands as an alternative for arsenic removal from reverse osmosis effluent.

In Argentina, drinking water for c.a. 10% of the population has arsenic (As) concentrations higher than those recommended by WHO (10 µg L-1). Reverse osmosis (RO) appears as an immediate and effective solution for As remediation. However, this process has a residual flow known as "rejection" or "concentrate" where dissolved species are more concentrated than in the feed flow. In this study, phytoremediation with subsurface horizontal-flow constructed wetlands (CW) was proposed to reduce As concentration in the RO residues. Experiments were carried out during 419 days at room temperature and using a continuous regime (flow of 36 L d-1, As concentration around 85 µg L-1) of RO rejection from a water treatment plant located in Buenos Aires province, Argentina. The study was performed using prototypes planted with Cyperus haspan (PA), Juncus effusus (PB) and a mix of inert gravel and laterite (substrate) that was used as a control (PC). Results showed that after a stabilization time, As removal (%) was between 30% and 80% in the CW planted with J. effusus and between 10 and 40% with C. haspan. As concentration along CW showed similarities between the prototypes PC and PA. The cumulative mass of As was 62%, 34% and 27% for PA, PB and PC, respectively. The contribution of C. haspan and J. effusus during the experimental time was between 12 and 67% and 22 to 87%, respectively. The bioaccumulation and translocation factors indicated that for J. effusus the accumulation is more important than the translocation process (1.6 and 0.2, respectively), while for C. haspan both factors were similar (1.1 and 1.0, respectively). Results suggested that this technology has the potential for an efficient and environmentally sustainable alternative to RO rejection treatment and disposal regarding As concentration.

Linomide prevents the lethal effect of anti-Fas antibody and reduces Fas-mediated ceramide production in mouse hepatocytes.

Fas is an apoptosis-signaling receptor molecule expressed in vivo on thymocytes, liver, heart, and ovary. In vivo administration of the anti-Fas Jo2 antibody in mice induces severe apoptotic liver damage leading to fulminant hepatitis and death. Linomide, a quinoline 3-carboxamide, inhibits apoptosis of B and T cells induced by various stimuli including viruses, superantigens, and glucocorticoids. Mice treated with linomide survived the lethal effect of anti-Fas antibody, did not accumulate ceramide in hepatocytes, and recovered liver structure and function within 96 h of anti-Fas injection, as confirmed by histology and glutamic oxalacetic transaminase, glutamic pyruvic transaminase, and lactate dehydrogenase levels. Surviving mice showed severe depletion of cortical thymocytes, but medullar thymic cells expressing high CD3 and Fas levels also survived the treatment with anti-Fas in the presence of linomide. Heart, lung, and ovary showed no signs of apoptosis promoted by Fas ligation. These results suggest that linomide prevents cell death triggered by Fas ligation and can be useful for therapeutic intervention in fulminant hepatitis.

Thymic selection.

The ability of T cells to recognize foreign antigens, distinguish them from self antigens, and regulate immune responses depends largely on which of the vast array of different T-cell receptors they display on their surface, the so called T-cell repertoire. Two selection processes, positive and negative selection, operate on developing T cells and allow only a subset of all possible T cells to differentiate. New developments in transgenic mouse technology and the study of T-cell signalling are unravelling these fascinating processes.

Apoptosis, Fas and systemic autoimmunity: the MRL-lpr/lpr model.

Proteins encoded by the fas and fas ligand (fasL) genes are involved in apoptotic cell death in lymphocytes. In this article we review the recent elucidation of the role of the Fas-FasL interactions in the maintenance of tolerance to self antigens and in the homeostatic regulation of lymphocyte clonal expansion, and discuss the mechanisms of autoimmunity in Fas- and FasL-deficient mutant mouse strains.

Evaluation of exercise testing for subjects with isolated proteinuria and/or hematuria of unclear significance.

BACKGROUND: A frequent problem that family doctors face is the meaning of small quantities of blood or protein in urine samples. Patients with this problem are often either neglected or referred to specialists for complex, expensive, and often invasive diagnostic procedures. Exercise testing has never been considered in nephrology, except for some attempts in diabetic patients. METHODS: We report on a study conducted over 12 years with patients referred for slight hematuria and/or proteinuria to determine whether exercise testing could be a diagnostic aid in some or all of them. We performed exercise testing using a treadmill preceded and followed by urine analysis, with a kidney biopsy within 10 days. Of the 94 patients enrolled in the study, only those with a positive exercise test turned out to have parenchymal nephropathy. At the end of the study, we simplified the quantification of exertion, dispensing with the treadmill and drastically reducing the number of urinary parameters considered. RESULTS: In patients with histological evidence of kidney damage, most of the variables increased significantly after the test. Statistical analysis also showed that determination of proteinuria and hematuria alone guaranteed maximum predictability. We found that it is also possible to simplify the quantification of effort/exertion and to drastically reduce the number of urinary parameters and still obtain significant results. CONCLUSIONS: Exercise testing provides useful information about the significance of microhematuria and proteinuria, reducing the number of cases that need to be referred to specialists. The method needs to be validated in other studies, but our results suggest that family doctors could use simple dipsticks to screen the many cases of microhematuria or proteinuria observed in daily practice. The method seems useful in eliminating doubts and unnecessary diagnostic costs.

The anatomy and vascularity of the lunate: considerations applied to Kienböck's disease.

PURPOSE: The purpose of this study was to assess the anatomy and vascularity of the lunate. The genesis of lunatomalacia requires some combination of vascular risk and mechanical predisposition. The findings will be correlated with the major existing theories of the cause of Kienböck's disease. METHODS: We studied 27 cadaver upper limbs using latex injection and the Spalteholz technique. We investigated the blood supply to the lunate. In 24 wrists we evaluated the incidence and distribution of anatomic features, arthrosis, and soft tissue lesions. We correlated the lunate morphology and ligaments disruptions with the arthritic changes. RESULTS: The lunate morphology results as classified by Antuña-Zapico were five type I (20.8%), 18 type II (75%) and one type III (4.2%). The lunate was found to have a separate facet for the hamate in 11 cases (45.8%). The most common size of the facet was found to be 3 mm (range, 3-6 mm). Arthrosis was identified with most frequency in the radius (88.2%) and lunate (94.1%). The triangular fibrocartilage complex was found torn in 58.3%, the lunotriquetral interosseous ligament was torn in 20.8% and the scapholunate interosseous ligament (SLIL) was torn in 54.2% of the wrists. There was a correlation between the presence of arthrosis at the hamate and the presence of a lunate facet (P=0.027) and a correlation between the presence of a tear in the SLIL and arthrosis in the scaphoid (P=0.002). The nutrient vessels entered the lunate through the dorsal and volar poles in all the specimens. The dorsal intercarpal and radiocarpal arches supply blood to the lunate from a plexus of vessels located directly over the lunate's dorsal pole. Vessels entered the dorsal aspect of the lunate through one to three foramina. One to five nutrient vessels were observed entering the volar pole through various ligament insertions, including the ligament of Testut-Kuentz (radio-scapho-lunate (RSL) ligament) and the radiolunate triquetrum ligament (or dorsoradial carpal ligament) and ulnar lunate triquetral ligament. CONCLUSIONS: The lunate had consistent dorsal and palmar arteries entering the bone in all the specimens. The blood supply and foramina number is greater in the volar pole of the lunate than the dorsal pole. The lunate blood supply comes from different ligaments. In the etiopathogeny of Kienböck's disease it is possible that an acute or chronic, traumatic or non-traumatic injury of the vessel bearing ligaments, particularly because of their structure and the location of the RSL ligament, may have an important role in the appearance of lunate necrosis.

[Endoscopic anatomy of the axillary brachial plexus: examination of cadavers and implications for anesthesia]. / Anatomía endoscópica del plexo braquial a nivel axilar. Estudio en cadáveres. Implicaciones anestésicas.

OBJECTIVE: To identify the structures of the brachial plexus using fiberoptic techniques, determine the most useful approach, and visualize the introduction of instruments used in anesthesia. MATERIAL AND METHODS: Formalin-treated a ndcryopreserved cadavers were studied with a 4-mm fiberoptic endoscope, an arthroscope for small joints, and nerve-stimulation needles through which a catheter could be introduced. The arm and elbow were dissected to isolate the humeral artery and the medial, radial, and cubital nerves. The fiberoptic endoscope was advanced along each nerve until the axilla, where the neurovascular structures of the plexus were identified. The axilla was punctured with a nerve stimulation needle and the catheter was passed through the needle while its movement was observed with the fiberoptic endoscope. Then, the axilla was punctured with the arthroscope as if it were the needle. RESULTS: In cryopreserved and formalin-treated cadavers, the fiberoptic endoscope could be passed as far as the axilla by following the medial nerve but not by following the cubital or radial nerves. We were able to identify the humeral artery and medial nerve in the axilla and to observe entry of the needle and progression of the catheter. The arthroscope could be introduced in the axilla as if it were a needle and the humeral artery and medial nerve could be identified. CONCLUSIONS: The inside of the neurovascular bundle of the brachial plexus can be visualized using afiberoptic endoscope advanced from the elbow or using an arthroscope introduced in the axilla. Although we are a long way from being able to see what we are doing, this is a first step towards direct visualization of the major structures of the axillary brachial plexus.

SPROUTY-2 represses the epithelial phenotype of colon carcinoma cells via upregulation of ZEB1 mediated by ETS1 and miR-200/miR-150.

SPROUTY-2 (SPRY2) is a modulator of tyrosine kinase receptor signaling with receptor- and cell type-dependent inhibitory or enhancing effects. Studies on the action of SPRY2 in major cancers are conflicting and its role remains unclear. Here we have dissected SPRY2 action in human colon cancer. Global transcriptomic analyses show that SPRY2 downregulates genes encoding tight junction proteins such as claudin-7 and occludin and other cell-to-cell and cell-to-matrix adhesion molecules in human SW480-ADH colon carcinoma cells. Moreover, SPRY2 represses LLGL2/HUGL2, PATJ1/INADL and ST14, main regulators of the polarized epithelial phenotype, and ESRP1, an epithelial-to-mesenchymal transition (EMT) inhibitor. A key action of SPRY2 is the upregulation of the major EMT inducer ZEB1, as these effects are reversed by ZEB1 knock-down by means of RNA interference. Consistently, we found an inverse correlation between the expression level of claudin-7 and those of SPRY2 and ZEB1 in human colon tumors. Mechanistically, ZEB1 upregulation by SPRY2 results from the combined induction of ETS1 transcription factor and the repression of microRNAs (miR-200 family, miR-150) that target ZEB1 RNA. Moreover, SPRY2 increased AKT activation by epidermal growth factor, whereas AKT and also Src inhibition reduced the induction of ZEB1. Altogether, these data suggest that AKT and Src are implicated in SPRY2 action. Collectively, these results show a tumorigenic role of SPRY2 in colon cancer that is based on the dysregulation of tight junction and epithelial polarity master genes via upregulation of ZEB1. The dissection of the mechanism of action of SPRY2 in colon cancer cells is important to understand the upregulation of this gene in a subset of patients with this neoplasia that have poor prognosis.

Role of the autophosphorylation site on the biological function of pp56lck.

Src-family tyrosine kinases act as signaling molecules in a wide array of cellular activation processes. The existence of the various src-family members reflects the requirement for different cell-surface receptors to transmit cell-type specific intracellular signals. The structural basis for the functional specificity of src-kinases is being actively investigated. In the present report we have analysed the contribution of the area surrounding the autophosphorylation site (located at subdomain VII of the catalytic domain) in determining src-kinases activity and functional specificity. To this end we analysed the kinase activities of the lymphoid src-kinase pp56lck and a mutant of pp56lck in which this region has been exchanged for the corresponding area of the serine/threonine kinase c-Raf. Our studies indicate that the change at subdomain VII affected the ability of pp56lck to phosphorylate physiological substrates. Furthermore, when analysed in T cells, the mutant at subdomain VII failed to induce interleukin-2 production, a specific biological function of pp56lck. Thus, the area surrounding the autophosphorylation site of pp56lck plays a critical role in mediating its specific biological function.