Conditioned Medium from Human Uterine Cervical Stem Cells Regulates Oxidative Stress and Angiogenesis of Retinal Pigment Epithelial Cells.

INTRODUCTION: Retinal homeostasis is essential to avoid retinal pigment epithelium (RPE) damage resulting in photoreceptor death and blindness. Mesenchymal stem cells-based cell therapy could contribute to the maintenance of the retinal homeostasis. We have explored the effect of human uterine cervical stem cells (hUCESCs)-conditioned medium (hUCESC-CM) on RPE cells under oxidative stress condition. METHODS: ARPE-19 cells were treated with hydrogen peroxide (H2O2) in the presence or absence of hUCESC-CM. qRT-PCR and Western blot were used to evaluate the expression of oxidative stress-related (HO-1, GCLC, and HSPB1) and vasculogenesis-related (VEGFA, PDGFA, and PDGFB) factors. Also, we assessed in vitro effects of hUCESC-CM on endothelial-cell (HUVEC) tube formation. RESULTS: mRNA expression of HO-1, GCLC, HSPB1, VEGFA, PDGFA, and PDGFB were significantly increased in ARPE-19 cells treated with H2O2 + hUCESC-CM compared to cells treated with H2O2 only. Regarding the tube formation assay, HUVEC treated with supernatant from ARPE-19 cells treated with H2O2 + hUCESC-CM showed a significant increase in average vessel length, number of capillary-like junctions, and average of vessels area compared with HUVEC treated with supernatant from ARPE-19 cells treated with H2O2 only. CONCLUSION: Our results show potential therapeutic effects of hUCESC-CM on RPE, such as protection from damage by oxidative stress, stimulation of detoxifying genes, and a better vascularization.

Anti-Inflammatory Effect of Tacrolimus/Hydroxypropyl-ß-Cyclodextrin Eye Drops in an Endotoxin-Induced Uveitis Model.

BACKGROUND: Uveitis is an infrequent disease which constitutes a major cause of ocular morbidity. Correct management is essential, being corticosteroids its cornerstone. In case of contraindication to corticosteroids or treatment failure, the use of topical tacrolimus (TAC) could be an alternative which has already demonstrated safety and effectiveness in other ocular pathologies. However, TAC eye drops are not marketed, thus their elaboration must be carried out in Hospital Pharmacy Departments (HPDs). METHODS: 32 Sprague-Dawley rats were divided into 4 groups of 8 rats each: (a) untreated healthy rats (Healthy); (b) untreated Endotoxin-Induced Uveitis model-rats (EIU); (c) EIU-rats treated with standard treatment of dexamethasone ophthalmic drops (DXM) and (d) EIU-rats treated with TAC-hydroxypropyl-ß-cyclodextrin eye drops previously developed by our group (TAC-HPßCD). The mRNA expression levels of IL-6, IL-8, MIP-1α and TNF-α, quantitative analysis of leucocytes in aqueous humor and histological evaluation were performed. RESULTS: TAC-HPßCD eye drops demonstrated to reduce ocular inflammation, expression of IL-6, TNF-α, MIP-1α and leukocyte infiltration in aqueous humor. CONCLUSIONS: TAC-HPßCD eye drops showed beneficial effect in EIU model in rats, positioning as an alternative for uveitis treatment in case of corticosteroids resistance or intolerance.

LIPG endothelial lipase and breast cancer risk by subtypes.

Experimental data showed that endothelial lipase (LIPG) is a crucial player in breast cancer. However, very limited data exists on the role of LIPG on the risk of breast cancer in humans. We examined the LIPG-breast cancer association within our population-based case-control study from Galicia, Spain, BREOGAN (BREast Oncology GAlicia Network). Plasma LIPG and/or OxLDL were measured on 114 breast cancer cases and 82 controls from our case-control study, and were included in the present study. The risk of breast cancer increased with increasing levels of LIPG (multivariable OR for the highest category (95% CI) 2.52 (1.11-5.81), P-trend = 0.037). The LIPG-breast cancer association was restricted to Pre-menopausal breast cancer (Multivariable OR for the highest LIPG category (95% CI) 4.76 (0.94-28.77), P-trend = 0.06, and 1.79 (0.61-5.29), P-trend = 0.372, for Pre-menopausal and Post-menopausal breast cancer, respectively). The LIPG-breast cancer association was restricted to Luminal A breast cancers (Multivariable OR for the highest LIPG category (95% CI) 3.70 (1.42-10.16), P-trend = 0.015, and 2.05 (0.63-7.22), P-trend = 0.311, for Luminal A and non-Luminal A breast cancers, respectively). Subset analysis only based on HER2 receptor indicated that the LIPG-breast cancer relationship was restricted to HER2-negative breast cancers (Multivariable OR for the highest LIPG category (95% CI) 4.39 (1.70-12.03), P-trend = 0.012, and 1.10 (0.28-4.32), P-trend = 0.745, for HER2-negative and HER2-positive tumors, respectively). The LIPG-breast cancer association was restricted to women with high total cholesterol levels (Multivariable OR for the highest LIPG category (95% CI) 6.30 (2.13-20.05), P-trend = 0.018, and 0.65 (0.11-3.28), P-trend = 0.786, among women with high and low cholesterol levels, respectively). The LIPG-breast cancer association was also restricted to non-postpartum breast cancer (Multivariable OR for the highest LIPG category (95% CI) 3.83 (1.37-11.39), P-trend = 0.003, and 2.35 (0.16-63.65), P-trend = 0.396, for non-postpartum and postpartum breast cancer, respectively), although we lacked precision. The LIPG-breast cancer association was more pronounced among grades II and III than grade I breast cancers (Multivariable ORs for the highest category of LIPG (95% CI) 2.73 (1.02-7.69), P-trend = 0.057, and 1.90 (0.61-6.21), P-trend = 0.170, for grades II and III, and grade I breast cancers, respectively). No association was detected for OxLDL levels and breast cancer (Multivariable OR for the highest versus the lowest category (95% CI) 1.56 (0.56-4.32), P-trend = 0.457).

Development and Characterization of a Tacrolimus/Hydroxypropyl-ß-Cyclodextrin Eye Drop.

Uveitis is a vision inflammatory disorder with a high prevalence in developing countries. Currently, marketed treatments remain limited and reformulation is usually performed to obtain a tacrolimus eye drop as a therapeutic alternative in corticosteroid-refractory eye disease. The aim of this work was to develop a mucoadhesive, non-toxic and stable topical ophthalmic formulation that can be safely prepared in hospital pharmacy departments. Four different ophthalmic formulations were prepared based on the tacrolimus/hydroxypropyl-ß-cyclodextrin (HPßCD) inclusion complexes' formation. Phase solubility diagrams, Nuclear Magnetic Resonance (NMR) and molecular modeling studies showed the formation of 1:1 and 1:2 tacrolimus/HPßCD inclusion complexes, being possible to obtain a 0.02% (w/v) tacrolimus concentration by using 40% (w/v) HPßCD aqueous solutions. Formulations also showed good ophthalmic properties in terms of pH, osmolality and safety. Stability studies proved these formulations to be stable for at least 3 months in refrigeration. Ex vivo bioadhesion and in vivo ocular permanence showed good mucoadhesive properties with higher ocular permanence compared to the reference pharmacy compounding used in clinical settings (t1/2 of 86.2 min for the eyedrop elaborated with 40% (w/v) HPßCD and Liquifilm® versus 46.3 min for the reference formulation). Thus, these novel eye drops present high potential as a safe alternative for uveitis treatment, as well as a versatile composition to include new drugs intended for topical ophthalmic administration.

Differential sensitivity of the On and Off visual responses to retinal ischemia.

Retinal ischemia is a common condition that may lead into vision impairment and blindness. In this study, we evaluated changes separately in On and Off visual responses induced by retinal ischemia. To do this, reversible retinal ischemia was induced in anaesthetized rats by increasing the intraocular pressure until the eye fundus became whitish for either 30 or 60 min. Both electroretinogram (ERG) and multiunit neuronal activity in the superior colliculus (SC) were recorded simultaneously for at least 20 min before, during, and after ischemia. In addition, in normal eyes, intravitreal glycine (Gly) injections were performed to further investigate the mechanisms involved in this process. We found that collicular Off responses were more sensitive to ischemia than On responses. The Off response was the first one to decay at the time ischemia was induced and the last to recover after blood reperfusion. The duration of ischemia also differentially affected both responses. After 30 min of ischemia, 14% of SC recordings failed to recover Off responses. After 1 h of ischemia, the percentage of recordings that failed to recover Off responses increased to 50%. Post-ischemic ERGs remained unaltered in all cases. Intravitreal Gly injections caused suppression of Off responses in the SC. Higher doses caused suppression of both On and Off responses in the SC but with no effect on the ERG at the doses tested. In summary, Off responses were more sensitive than On responses to ischemia suggesting that different mechanisms drive the two types of responses. The recovery of transitory ischemia was not complete in the SC responses whereas the ERG remained unaltered, suggesting that retinal damage produced by ischemia is more prominent in ganglion cells. Our results provide critical information for understanding ischemia repercussions and visual processing in the early visual system.

Using Herbs and Spices to Increase Vegetable Intake Among Rural Adolescents.

OBJECTIVE: To test whether adding herbs and spices to school lunch vegetables increases selection and intake compared with lightly salted control versions among rural adolescents. DESIGN: This study compared intake of vegetables with herbs and spices with lightly salted controls (phase I) and tested whether 5 repeated exposures would increase students' intake of herb and spice seasoned vegetables (phase II). PARTICIPANTS AND SETTING: A total of 600-700 students at a rural middle/high school (age 11-18 years). INTERVENTION: In phase I, herbs and spices were added to 8 vegetables and outcomes were compared with 8 control recipes. In phase II, the impact of repeated exposure to herb and spice blends served on different vegetables was assessed. MAIN OUTCOMES: Vegetable selection rates, weighed intake, and willingness to eat again. ANALYSIS: Two-way ANOVAs tested effects of condition (herbs and spices vs control; before vs after exposure) and age (middle vs high school) on selection and intake. RESULTS: In phase I, students ate more control than seasoned broccoli (P = .01), cauliflower (P = .006), and green beans (P = .01), and high schoolers generally consumed more seasoned vegetables than did middle schoolers (P < .03). In phase II, repeated exposure to herbs and spices increased reported willingness to eat again for seasoned broccoli (P = .003). CONCLUSIONS AND IMPLICATIONS: In a short-term intervention, herbs and spices did not produce robust increases in school lunch vegetable intake among rural adolescents, but limited repeat exposure may increase students' willingness to consume these flavors. Additional work is needed to identify individual and school-level characteristics that affect students' willingness to select and consume vegetables with herbs and spices.

Food commercials do not affect energy intake in a laboratory meal but do alter brain responses to visual food cues in children.

Food commercials promote snack intake and alter food decision-making, yet the influence of exposure to food commercials on subsequent neural processing of food cues and intake at a meal is unclear. This study tested whether exposing children to food or toy commercials altered subsequent brain response to high- and low-energy dense food cues and influenced laboratory intake at a multi-item, ad libitum meal. Forty-one 7-9-year-old children (25 healthy weight; 16 with overweight/obesity) completed five visits as part of a within-subjects design where they consumed multi-item test-meals under three conditions: no exposure, food commercial exposure, and toy commercial exposure. On the fourth and fifth visits, functional magnetic resonance imaging (fMRI) was performed while children viewed low- and high-energy dense food images following exposure to either food or toy commercials. Linear mixed models tested for differences in meal energy intake by commercial condition. A whole-brain analysis was conducted to compare differences in response by commercial condition and child weight status. Meal intake did not differ by commercial condition (p = 0.40). Relative to toy commercials, food commercials reduced brain response to high-energy food stimuli in cognitive control regions, including bilateral superior temporal gyri, middle temporal gyrus, and inferior frontal gyrus. Commercial condition * weight status interactions were observed in orbitofrontal cortex, fusiform gyrus, and supramarginal gyrus. Children with overweight/obesity showed increased response in these regions to high-energy stimuli following food commercials. Food commercial exposure affected children's subsequent processing of food cues by reducing engagement of the prefrontal cortex, a region implicated in cognitive control. Even though food commercial exposure did not increase intake at a meal, the effect of reduced prefrontal cortical engagement on a broader range of consumption patterns warrants investigation.

Corneal regeneration by conditioned medium of human uterine cervical stem cells is mediated by TIMP-1 and TIMP-2.

The aim of the present study was to evaluate the effect and the mechanism of action of the conditioned medium from human uterine cervical stem cells (CM-hUCESC) on corneal wound healing in a rabbit dry eye model. To do this, dry eye and corneal epithelial injuries were induced in rabbits by topical administration of atropine sulfate and NaOH. Hematoxylin-Eosin (H&E) and Ki-67 immunostaining were carried out to evaluate corneal damage and cell proliferation, and real-time PCR was used to evaluate proinflammatory cytokines in the cornea. In addition, in order to investigate possible factors involved in corneal regeneration, primary cultures of rat corneal epithelial cells (rCECs) were used to evaluate cell migration, proliferation, and apoptosis before and after immunoprecipitation of specific factors from the CM-hUCESC. Results showed that CM-hUCESC treatment significantly improved epithelial regeneration in rabbits with dry eye induced by atropine and reduced corneal pro-inflammatory TNF-α, MCP-1, MIP-1α and IL-6 cytokines. In addition, metalloproteinase inhibitors TIMP-1 and TIMP-2, which are present at high levels in CM-hUCESC, mediated corneal regenerative effects by both inducing corneal epithelial cell proliferation and inhibiting apoptosis. In summary, CM-hUCESC induces faster corneal regeneration in a rabbit model of dry eye induced by atropine than conventional treatments, being TIMP-1 and TIMP-2 mediators in this process. The results indicate that an alternative CM-based treatment for some corneal conditions is achievable, although future studies would be necessary to investigate other factors involved in the multiple observed effects of CM-hUCESC.

Representaciones sociales de adolescentes mexicanas embarazadas sobre el puerperio, la lactancia y los cuidados del recién nacido / Social representations of Mexican pregnant teenagers about the puerperal care, lactation, and newborn care

Resumen: Introducción: Los cuidados durante el puerperio y la alimentación del recién nacido están guiados por significados culturales afianzados entre las mujeres, por lo que es importante conocerlos e identificar cómo se adquieren y perpetúan. En este tenor, se estudiaron las representaciones sociales que tienen adolescentes mexicanas embarazadas sobre el puerperio, la lactancia y el cuidado del recién nacido. Métodos: Se realizó un estudio interpretativo basado en los principios de la teoría de las representaciones sociales. Se realizaron entrevistas para obtener información de 30 adolescentes mexicanas que asistieron a control prenatal al área de ginecoobstetricia en un hospital de segundo nivel de atención durante 2015. Se aplicaron estrategias de análisis de contenido clásico para analizar la información; este proceso consistió en codificar y categorizar la información. También se elaboró un mapa conceptual para describir las representaciones sociales encontradas. Resultados: Se identificaron 190 códigos y tres representaciones sociales: «lactancia: práctica mitificada¼, «los recién nacidos son frágiles¼ y «madre e hijo deben sincronizarse¼. Conclusiones: Se identificaron tres representaciones sociales que explicaron las prácticas de las adolescentes hacia la lactancia y el cuidado de ellas y sus hijos, adquiridas mediante la comunicación familiar y afianzadas por la necesidad de apoyo debido a ausencia temporal o permanente de pareja, crisis personales motivadas por los cambios corporales, miedo a sufrir cambios por la lactancia y desconocimiento sobre cómo llevar a cabo la lactancia y los cuidados durante el puerperio.

Abstract: Background: Puerperal care and feeding of the newborn are guided by entrenched cultural meanings between women, so it is important to know and identify how they are acquired and perpetuated. Regarding this knowledge, the social representations that Mexican pregnant teenagers have about puerperium, lactation and newborn care were studied. Methods: An interpretative study was made based on principles of the theory of social representations. Interviews were conducted to obtain information from 30 Mexican adolescents who attended prenatal care at the gynecological obstetrics area in a second-level hospital during 2015. Classical content analysis strategies were applied to analyze the information; this process consisted of coding and categorizing information. A conceptual map was also developed to describe the social representations found. Results: In this study, 190 codes and three social representations were identified: "breastfeeding is a practice based on myths", "newborns are fragile" and "mother and child must be synchronized". Conclusions: Three social representations were identified that explain the practices of adolescents towards breastfeeding and the care of them and their children, which were acquired through family communication and strengthened by the need for support due to the temporary or permanent absence of the couple, personal crises motivated by bodily changes, fear of new modifications due to breastfeeding and ignorance about how carry out breastfeeding and care during the puerperium.

[Social representations of Mexican pregnant teenagers about the puerperal care, lactation, and newborn care]. / Representaciones sociales de adolescentes mexicanas embarazadas sobre el puerperio, la lactancia y los cuidados del recién nacido.

Background: Puerperal care and feeding of the newborn are guided by entrenched cultural meanings between women, so it is important to know and identify how they are acquired and perpetuated. Regarding this knowledge, the social representations that Mexican pregnant teenagers have about puerperium, lactation and newborn care were studied. Methods: An interpretative study was made based on principles of the theory of social representations. Interviews were conducted to obtain information from 30 Mexican adolescents who attended prenatal care at the gynecological obstetrics area in a second-level hospital during 2015. Classical content analysis strategies were applied to analyze the information; this process consisted of coding and categorizing information. A conceptual map was also developed to describe the social representations found. Results: In this study, 190 codes and three social representations were identified: "breastfeeding is a practice based on myths", "newborns are fragile" and "mother and child must be synchronized". Conclusions: Three social representations were identified that explain the practices of adolescents towards breastfeeding and the care of them and their children, which were acquired through family communication and strengthened by the need for support due to the temporary or permanent absence of the couple, personal crises motivated by bodily changes, fear of new modifications due to breastfeeding and ignorance about how carry out breastfeeding and care during the puerperium.

Introducción: Los cuidados durante el puerperio y la alimentación del recién nacido están guiados por significados culturales afianzados entre las mujeres, por lo que es importante conocerlos e identificar cómo se adquieren y perpetúan. En este tenor, se estudiaron las representaciones sociales que tienen adolescentes mexicanas embarazadas sobre el puerperio, la lactancia y el cuidado del recién nacido. Métodos: Se realizó un estudio interpretativo basado en los principios de la teoría de las representaciones sociales. Se realizaron entrevistas para obtener información de 30 adolescentes mexicanas que asistieron a control prenatal al área de ginecoobstetricia en un hospital de segundo nivel de atención durante 2015. Se aplicaron estrategias de análisis de contenido clásico para analizar la información; este proceso consistió en codificar y categorizar la información. También se elaboró un mapa conceptual para describir las representaciones sociales encontradas. Resultados: Se identificaron 190 códigos y tres representaciones sociales: «lactancia: práctica mitificada¼, «los recién nacidos son frágiles¼ y «madre e hijo deben sincronizarse¼. Conclusiones: Se identificaron tres representaciones sociales que explicaron las prácticas de las adolescentes hacia la lactancia y el cuidado de ellas y sus hijos, adquiridas mediante la comunicación familiar y afianzadas por la necesidad de apoyo debido a ausencia temporal o permanente de pareja, crisis personales motivadas por los cambios corporales, miedo a sufrir cambios por la lactancia y desconocimiento sobre cómo llevar a cabo la lactancia y los cuidados durante el puerperio.

Visual stimulation quenches global alpha range activity in awake primate V4: a case study.

Increasing evidence suggests that sensory stimulation not only changes the level of cortical activity with respect to baseline but also its structure. Despite having been reported in a multitude of conditions and preparations (for instance, as a quenching of intertrial variability, Churchland et al., 2010), such changes remain relatively poorly characterized. Here, we used optical imaging of voltage-sensitive dyes to explore, in V4 of an awake macaque, the spatiotemporal characteristics of both visually evoked and spontaneously ongoing neuronal activity and their difference. With respect to the spontaneous case, we detected a reduction in large-scale activity ([Formula: see text]) in the alpha range (5 to 12.5 Hz) during sensory inflow accompanied by a decrease in pairwise correlations. Moreover, the spatial patterns of correlation obtained during the different visual stimuli were on the average more similar one to another than they were to that obtained in the absence of stimulation. Finally, these observed changes in activity dynamics approached saturation already at very low stimulus contrasts, unlike the progressive, near-linear increase of the mean raw evoked responses over a wide range of contrast values, which could indicate a specific switching in the presence of a sensory inflow.

Preclinical PET Study of Intravitreal Injections.

Purpose: This work aimed at describing the time course of vitreous clearance through the use of positron emission tomography (PET) as a noninvasive tool for pharmacokinetic studies of intravitreal injection. Methods: The pharmacokinetic profile of intravitreal injections of molecules labeled with 18Fluorine (18F) was evaluated in adult Sprague Dawley rats by using a dedicated small-animal PET/computed tomography scanner. Different conditions were studied: three molecules radiolabeled with 18F (18F-FDG, 18F-NaF, and 18F-Choline), three volumes of intravitreal injections (7, 4, and 2 µL), and absence or presence of eye inflammation (uveitis). Results: Our results showed that there are significant pharmacokinetic differences among the radiolabeled molecules studied but not among the injected volumes. The presence or absence of uveitis was an important factor in vitreous clearance, since the elimination of the drug was clearly increased when this condition is present. Conclusions: Intravitreal pharmacokinetic studies based on the use of dedicated PET imaging can be of potential interest as noninvasive tools in ophthalmic drug development in small animals.

Anti-inflammatory effect of conditioned medium from human uterine cervical stem cells in uveitis.

The aim of the present study was to evaluate the effect of conditioned medium from human uterine cervical stem cells (CM-hUCESCs) in uveitis. To do that, uveitis was induced in rats after footpad injection of Escherichia coli lipopolysaccaride (LPS). Human retinal pigment epithelial (ARPE-19) cells after LPS challenge were used to test anti-inflammatory effect of CM-hUCESCs 'ìn vitro'. Real-time PCR was used to evaluate mRNA expression levels of the pro-inflammatory cytokines interkeukin-6, interkeukin-8, macrophage inflammatory protein-1 alpha, tumor necrosis factor alpha, and the anti-inflammatory interkeukin-10. Leucocytes from aqueous humor (AqH) were quantified in a Neubauer chamber, and eye histopathological analysis was done with hematoxylin-eosin staining. Additionally, using a human cytokine antibody array we evaluated CM-hUCESCs to determine mediating proteins. Results showed that administration of CM-hUCESCs significantly reduced LPS-induced pro-inflammatory cytokines both 'in vitro' and 'in vivo', and decreased leucocytes in AqH and ocular tissues. High levels of cytokines with anti-inflammatory effects were found in CM-hUCESCs, suggesting a possible role of these factors in reducing intraocular inflammation. In summary, treatment with CM-hUCESCs significantly reduces inflammation in uveitis. Our data indicate that CM-hUCESCs could be regarded as a potential therapeutic agent for patients suffering from ocular inflammation.

Biological evaluation of new vitamin D2 analogues.

1,25-dihydroxyvitamin D3 (1,25D), a steroid hormone which regulates calcium/phosphate homeostasis, has a broad spectrum of anti-cancer activities, including differentiation of acute myeloid leukemia (AML) cells. In order to avoid undesirable side effects such as hypercalcemia, low-calcemic analogues should be produced for therapeutic purposes. In this paper, we describe biological activities of double-point modified analogues of vitamin D2 and we compare them to 1,25D and to paricalcitol, the drug used to treat secondary hyperparathyroidism. In vivo, our new analogues have lower calcemic effects, and lower toxicity in comparison to 1,25D. They have enhanced pro-differentiating and transcription-inducing activities in AML cells. Interestingly, differentiation effects do not correlate with the affinities of the analogues to the vitamin D receptor (VDR).

Visual Perception in Anterior Temporal Lobectomy.

PURPOSE: To study visual perception in patients with anterior temporal lobectomy. MATERIALS AND METHODS: We explored some aspects of visual perception and compared the results obtained from 14 control subjects and 14 patients with unilateral anterior temporal lobectomy. Each group included 7 men and 7 women and the same age distribution (patients and controls: age range 27-48 years; mean 37 years). All subjects underwent a conventional ophthalmic examination and were tested for color perception, stereopsis, texture perception, face recognition, and visual illusions. To quantify color, stereoscopic, and texture perception they performed a visuomotor task that required a rapid response to a visual stimulus. Reaction times were measured under several conditions. RESULTS: Mild visual field defects involving the superior quadrant contralateral to the lobectomy were found in five patients; two other patients presented more severe defects. Lobectomized patients showed a lower number of correct trials than normal subjects when performing tasks involving color and texture perception. These patients also had longer reaction times for color, stereoscopic, and texture stimulus detection. Face recognition and perception of illusory images were preserved after unilateral anterior temporal lobectomy. CONCLUSION: Our data indicate that patients with anterior temporal lobectomy show moderate deficits in color, stereo, and texture perception, with no impairment in complex visual stimuli perception.

Neural activity in monkey amygdala during performance of a multisensory operant task.

In this paper, we study the potential involvement of monkey amygdala in the evaluation of value encoding of visual and auditive stimuli associated with reward or no reward. We recorded the activity of 93 extracellular neurons from the monkey right amygdala, while performing a multisensory operant task. The activity of 78 task-related neurons was studied. Of these, 13 neurons (16%) responded to the value of visual stimuli, 22 neurons (28%) responded after the presentation of visual stimuli, 22 neurons (28%) showed an inhibition around the lever-pressing and were classified as action related neurons and 22 neurons (28%) responded after reward delivery. These findings suggest that neurons in the amygdala play a role in encoding value and processing visual information, participate in motor regulation and are sensitive to reward. The activity of these neurons did not change in the evaluation of auditive stimuli. These data support the hypothesis that amygdala neurons are specific to each sensory modality and that different groups of amygdala neurons process visual and auditive information.

Corneal epithelial wound healing and bactericidal effect of conditioned medium from human uterine cervical stem cells.

PURPOSE: To evaluate the effect of conditioned medium from human uterine cervical stem cells (CM-hUCESCs) on corneal epithelial healing in a rat model of dry eye after alkaline corneal epithelial ulcer. We also tested the bactericidal effect of CM-hUCESCs. METHODS: Dry eye was induced in rats by extraocular lacrimal gland excision, and corneal ulcers were produced using NaOH. Corneal histologic evaluation was made with hematoxylin-eosin (H&E) staining. Real-time PCR was used to evaluate mRNA expression levels of proinflammatory cytokines. We also studied the bactericidal effect of CM-hUCESCs in vitro and on infected corneal contact lenses (CLs) using Escherichia coli and Staphylococcus epidermidis bacteria. In addition, in order to investigate proteins from CM-hUCESCs that could mediate these effects, we carried out a human cytokine antibody array. RESULTS: After injury, dry eyes treated with CM-hUCESCs significantly improved epithelial regeneration and showed reduced corneal macrophage inflammatory protein-1 alpha (MIP-1α) and TNF-α mRNA expression as compared to untreated eyes and eyes treated with culture medium or sodium hyaluronate ophthalmic drops. In addition, we found in CM-hUCESCs high levels of proteins, such as tissue inhibitors of metalloproteinases 1 and 2, fibroblast growth factor 6 and 7, urokinase receptor, and hepatocyte growth factor, that could mediate these effects. In vitro, CM-hUCESCs showed a clear bactericidal effect on both E. coli and S. epidermidis and CLs infected with S. epidermidis. Analyses of CM-hUCESCs showed elevated levels of proteins that could be involved in the bactericidal effect, such as the chemokine (C-X-C motif) ligands 1, 6, 8, 10, and the chemokine (C-C motif) ligands 5 and 20. CONCLUSIONS: Treatment with CM-hUCESCs improved wound healing of alkali-injured corneas and showed a strong bactericidal effect on CLs. Patients using CLs and suffering from dry eye, allergies induced by commercial solutions, or small corneal injuries could benefit from this treatment.

Cancer progression by breast tumors with Pit-1-overexpression is blocked by inhibition of metalloproteinase (MMP)-13.

INTRODUCTION: The POU class 1 homeobox 1 transcription factor (POU1F1, also known as Pit-1) is expressed in the mammary gland and its overexpression induces profound phenotypic changes in proteins involved in cell proliferation, apoptosis, and invasion. Patients with breast cancer and elevated expression of Pit-1 show a positive correlation with the occurrence of distant metastasis. In this study we evaluate the relationship between Pit-1 and two collagenases: matrix metalloproteinase-1 (MMP-1) and matrix metalloproteinase-13 (MMP-13), which have been related to metastasis in breast cancer. METHODS: We began by transfecting the MCF-7 and MDA-MB-231 human breast adenocarcinoma cell lines with the Pit-1 overexpression vector (pRSV-hPit-1). Afterward, the mRNA, protein, and transcriptional regulation of both MMP-1 and MMP-13 were evaluated by real-time PCR, Western blot, chromatin immunoprecipitation (ChIP), and luciferase reporter assays. We also evaluated Pit-1 overexpression with MMP-1 and MMP-13 knockdown in a severe combined immunodeficiency (SCID) mouse tumor xenograft model. Finally, by immunohistochemistry we correlated Pit-1 with MMP-1 and MMP-13 protein expression in 110 human breast tumors samples. RESULTS: Our data show that Pit-1 increases mRNA and protein of both MMP-1 and MMP-13 through direct transcriptional regulation. In SCID mice, knockdown of MMP-13 completely blocked lung metastasis in Pit-1-overexpressing MCF-7 cells injected into the mammary fat pad. In breast cancer patients, expression of Pit-1 was found to be positively correlated with the presence of both MMP-1 and MMP-13. CONCLUSIONS: Our data indicates that Pit-1 regulates MMP-1 and MMP-13, and that inhibition of MMP-13 blocked invasiveness to lung in Pit-1-overexpressed breast cancer cells.

Potential therapeutic effect of the secretome from human uterine cervical stem cells against both cancer and stromal cells compared with adipose tissue stem cells.

Evidences indicate that tumor development and progression towards a malignant phenotype depend not only on cancer cells themselves, but are also deeply influenced by tumor stroma reactivity. The present study uses mesenchymal stem cells from normal human uterine cervix (hUCESCs), isolated by the minimally invasive method of routine Pap cervical smear, to study their effect on the three main cell types in a tumor: cancer cells, fibroblasts and macrophages. Administration of hUCESCs-conditioned medium (CM) to a highly invasive breast cancer MDA-MB-231 cell line and to human breast tumors with high cell proliferation rates had the effect of reducing cell proliferation, modifying the cell cycle, inducing apoptosis, and decreasing invasion. In a xenograft mouse tumor model, hUCESCs-CM reduced tumor growth and increased overall survival. In cancer-associated fibroblasts, administration of hUCESCs-CM resulted in reduced cell proliferation, greater apoptosis and decreased invasion. In addition, hUCESCs-CM inhibited and reverted macrophage differentiation. The analysis of hUCESCs-CM (fresh and lyophilized) suggests that a complex paracrine signaling network could be implicated in the anti-tumor potential of hUCESCs. In light of their anti-tumor potential, the easy cell isolation method, and the fact that lyophilization of their CM conserves original properties make hUCESCs good candidates for experimental or clinical applications in anticancer therapy.