α-Synuclein Drives Tau's Cytotoxic Aggregates Formation through Hydrophobic Interactions.

Tau and α-synuclein are proteins involved in pathologies known as tauopathies and synucleinopathies, respectively. Moreover, evidence shows that there is a crosstalk between them as is seen in the brains of individuals with sporadic neurodegenerative disorders. Based on that, we present data showing that the hydrophobic α-peptide 71 VTGVTAVAQKTV82 induces the aggregation of the full-length tau fragment in the absence of heparin assessed by ThT. Moreover, AFM images reveal the presence of straight filaments and amorphous aggregates of full-length tau in the presence of the α-peptide. Additionally, ITC experiments showed the interaction of the α-peptide with tau full-length (441 amino acids),4R (amino acids from 244 to 369), and both hexapeptides 275 VQIINK280 and 306 VQIVYK311 through hydrophobic interactions. The Raman spectroscopy spectra showed conformational changes in the Amide region in the aggregates formed with full-length tau and α-syn peptide. Furthermore, the incubation of extracellular aggregates with N2a cells showed morphological differences in the cellular body and the nucleus suggesting cell death. Moreover,, the incubation of different types of aggregates in cell culture provokes the release of Lactate dehydrogenase (LDH). Altogether, we found that α-synuclein peptide can drive the aggregation of full-length tau-provoking morphological and structural changes evoking cytotoxic effects.

Management of the axilla in postmenopausal patients with cN0 hormone receptor-positive/HER2-negative breast cancer treated with neoadjuvant endocrine therapy and its prognostic impact.

PURPOSE: To evaluate the differences in nodal positivity if the sentinel lymph node biopsy (SLNB) is performed before or after neoadjuvant endocrine therapy (NET) in breast cancer patients, and its impact on prognosis. METHODS: A retrospective cohort study was performed in a single center including 91 postmenopausal cases with clinically node-negative and hormone receptor-positive/HER2-negative (HR + /HER2-) breast cancer, treated with NET and SLNB. SLNB was done pre-NET until 2014, and post-NET thereafter. Axillary lymph node dissection (ALND) was indicated only in SLNB macrometastasis, although in selected elderly patients, it was omitted. Kaplan-Meier survival curves were estimated in relation to the status of the axilla, and the differences assessed using the log-rank test. RESULTS: Between December 2006 and March 2022, SLNB was performed pre-NET in 14 cases and post-NET in 77. Both groups were similar in baseline tumor and patient characteristics. SLNB positivity was similar regardless of whether SLNB was performed before (5/14, 35.7%) or after NET (27/77, 37%), with 2/14 SLN macrometastases in the pre-NET cohort and 17/77 in the post-NET cohort. Only three patients (18.7%) with SLN macrometastasis had > 3 positive nodes following ALND. The 5-year overall survival and distant disease-free survival were 92.4% and 94.8%, respectively, with no significant differences according to SLNB status (p 0.5 and 0.8, respectively). CONCLUSION: SLN positivity did not differ according to its timing (before or after NET). Therefore, NET has no effect on lymph node clearance. Furthermore, the prognosis is good regardless of the axillary involvement. Therefore, factors other than axillary involvement may affect the prognosis in these patients.

Impact of the initial experience in the penile inversion vaginoplasty technique on satisfaction levels: a pilot retrospective cohort study.

Background: There is a growing interest in reporting satisfaction levels of transgender women undergoing vaginoplasty surgery. The lack of information regarding satisfaction during the initial experience of the vaginoplasty technique, and the moderate morbidity related to the surgery, could discourage the immersion of new groups in initiating a program of this kind. Therefore, we aim to report patients' level of satisfaction during our initial experience in the penile inversion vaginoplasty technique. Methods: Retrospective study of patients who underwent penile inversion vaginoplasty in our center between September 2019 and August 2021. Surgery technique, demographic data, preoperative clinical variables, and short and long-term follow-up are described. Six months after surgery, a survey elaborated by the research team was conducted by phone. The score goes from 1 to 5, and it evaluates satisfaction on esthetics, functional, psychosocial, and global aspects. Results: Twenty patients underwent penile inversion vaginoplasty in our center during the described period. The average age was 35.6 years old, the mean body mass index (BMI) was 24.7 kg/m2, and they presented low comorbidity. Half of the patients presented at least one complication, most of which were minor. One patient was urgently reoperated due to bleeding, and three patients were reoperated on a scheduled basis from minor surgeries. 90% of the patients answered the questionnaire. The most common answers to all four areas covered (esthetics, functional, psychosocial, and global) were satisfied or very satisfied, resulting in a mean over four points in each one of the sections. Lastly, 94.4% of the patients reported being satisfied with their choice of having undergone surgery. Conclusions: Our initial experience in penile inversion vaginoplasty reveals good satisfaction results at short follow up.

The response of the mental health network of the Salamanca area to the COVID-19 pandemic: The role of the telemedicine.

The COVID-19 pandemic reached world-wide causing a great impact on healthcare services. The aim of this work is to describe the response of the Mental Health Network of the Salamanca´ Area (Spain) to this crisis and the reorganization of its resources within the first 8 weeks after the state of alarm was declared. The Psychiatry Service applied a contingency plan which included the reorganization of the human resources, the closure of some of the units and the implementation of telemedicine programs along with two specific programs, namely a mental health assistance program in the context of the infection by coronavirus, and another program for homeless people. 9.038 phone interviews were carried out in the outpatients and community mental health programs. The activity in subacute and acute wards, as well as that of the day hospital programs was decreased to 50%. Based on that this real-world response provided we concluded that the usage of telemedicine is promising in patients with any kind of disorder. Its implementation in daily practice will be considered in the future. Research must continue on COVID-19's impact on patients with mental disorders and Psychiatry's necessary adaptations and new approaches to them.

Current HHT genetic overview in Spain and its phenotypic correlation: data from RiHHTa registry.

BACKGROUND: Hereditary hemorrhagic telangiectasia (HHT) is a rare vascular disease with autosomal dominant inheritance. Disease-causing variants in endoglin (ENG) and activin A receptor type II-like 1 (ACVRL1) genes are detected in more than 90% of cases submitted to molecular diagnosis. METHODS: We used data from the RiHHTa (Computerized Registry of Hereditary Hemorrhagic Telangiectasia) registry to describe genetic variants and to assess their genotype-phenotype correlation among HHT patients in Spain. RESULTS: By May 2019, 215 patients were included in the RiHHTa registry with a mean age of 52.5 ± 16.5 years and 136 (63.3%) were women. Definitive HHT diagnosis defined by the Curaçao criteria were met by 172 (80%) patients. Among 113 patients with genetic test, 77 (68.1%) showed a genetic variant in ACVRL1 and 36 (31.8%) in ENG gene. The identified genetic variants in ACVRL1 and ENG genes and their clinical significance are provided. ACVRL1 mutations were more frequently nonsense (50%) while ENG mutations were more frequently, frameshift (39.1%). ENG patients were significantly younger at diagnosis (36.9 vs 45.7 years) and had pulmonary arteriovenous malformations (AVMs) (71.4% vs 24.4%) and cerebral AVMs (17.6% vs 2%) more often than patients with ACVRL1 variants. Patients with ACVRL1 variants had a higher cardiac index (2.62 vs 3.46), higher levels of hepatic functional blood tests, and anemia (28.5% vs 56.7%) more often than ENG patients. CONCLUSIONS: ACVRL1 variants are more frequent than ENG in Spain. ACVRL1 patients developed symptomatic liver disease and anemia more often than ENG patients. Compared to ACVRL1, those with ENG variants are younger at diagnosis and show pulmonary and cerebral AVMs more frequently.

[Sequencing and combination of agents in castration resistant prostate cancer.] / Secuenciación y combinación de agentes en cáncer de próstata resistente a la castración.

Prostate Cancer is the second most frequent malignant neoplasm in males in the world. At the end of the disease, when the tumor becomes resistant to castration, we have a wide range of treatment possibilities aimed at the Androgenic Receptor, androgens synthesis, the skeleton, chemotherapy, and even new molecular targets that are still under investigation. Today, the best sequence of treatment for each patient has not been established yet. OBJECTIVE: The objective of this work is to review the current scene of treatment in castrate resistant prostate cancer, as well as the latest developments and strategies to choose the best sequence in each patient. MATERIAL AND METHODS: A literature review was performed through Medline Database (Pubmed) using as key words: "Castrate Resistant Prostate Cancer", "Sequencing", "Biomarkers", "Systemic Therapy". We also reviewed ASCO GU 2017 abstracts. RESULTS: Since Docetaxel was approved in 2004, which increased overall survival by about 2 months in patients with Metastatic Castration Resistant Prostate Cancer, in recent years a large number of therapies have been approved, demonstrating an increase in overall survival after several phase III clinical trials: Cabacitaxel, Abiraterone, Enzalutamide, Sipuleucel-T, Denosumab, Radium 223. And more recently, some investigations about new targeted therapies directed to the androgen receptor, with greater affinity than enzalutamide, or more accurate inhibitors of CYP 17 enzyme than abiraterone, as well as, agents as monoclonal antibodies (anti PD1), vaccines, poly adenosine diphosphate- ribose polymerase inhibitors, are coming to the light. In the future, these outcomes could tune up the treatment sequencing, through the study of predictive biomarkers that will indicate the right target of each therapy. CONCLUSIONS: In the near future, outcomes of different clinical trials that are studying new molecules, will allow us to apply the sequencing of different therapies based on biomarkers present in blood (circulating tumor cells) or in specimen biopsies, achieving an increase in overall survival and improving quality of life of patients in the advanced stage of the disease, however the best choice of sequence is unknown at this moment.

Secuenciación y combinación de agentes en cáncer de próstata resistente a la castración / Sequencing and combination of agents in castration resistant prostate cancer

El cáncer de próstata (CP) es la segunda neoplasia maligna más frecuente en el mundo dentro del sexo masculino. Para la fase final de la enfermedad, cuando el tumor se hace resistente a castración tenemos un amplio abanico de posibilidades de tratamiento dirigidos al Receptor Androgénico, a la síntesis de andrógenos, al esqueleto, quimioterapia, incluso nuevas dianas moleculares que están aún en estudio. Aún así hoy en día no se ha establecido la secuencia idónea de tratamiento para cada paciente. Objetivos: El objetivo de este trabajo es plantear el panorama actual de tratamiento en la fase de resistencia a la castración y las últimas novedades y estrategias para elegir la mejor secuencia en el paciente adecuado. Métodos: Se ha realizado una búsqueda a través de Medline Database (Pubmed) usando como palabras Clave: "Castrate Resistant Prostate Cancer", "Sequencing", "Biomarkers", "Systemic Therapy". Revisión de abstract de ASCO GU 2017. Resultados: Desde que se aprobara Docetaxel en 2004 que aumentaba la supervivencia global unos 2 meses en pacientes afectos Cáncer de Próstata Resistente a la Castración Metastásicos, en los últimos años se han aprobado un buen número de terapias demostrando aumento de la supervivencia global tras ensayos clínicos en fase III: Cabacitaxel, abiraterona, enzalutamida, sipuleucel T, denosumab, Radium 223. Y más recientemente están saliendo a la luz investigaciones de nuevas terapias dirigidas al receptor androgénico, con una mayor afinidad que enzalutamida, o inhibidores de la enzima CYP 17 más precisos que abiraterona, así como anticuerpos monoclonales (anti PD1), vacunas, inhibidores de la poli adenosina difosfato-ribosa polimerasa, que en un futuro podrían afinar la secuencia de tratamiento de cada paciente, siendo necesario el estudio de biomarcadores predictores que indiquen la diana de cada terapia. Conclusiones: En un futuro próximo con los resultados de los distintos ensayos clínicos que estudian las nuevas moléculas podremos aplicar la secuenciación de las distintas terapias en función de biomarcadores presentes en la sangre (células tumorales circulantes) o en las biopsias, consiguiendo aumentar la supervivencia y la calidad de vida de los pacientes en la fase avanzada de la enfermedad, pero por el momento la mejor elección de secuencia es desconocida

Prostate Cancer is the second most frequent malignant neoplasm in males in the world. At the end of the disease, when the tumor becomes resistant to castration, we have a wide range of treatment possibilities aimed at the Androgenic Receptor, androgens synthesis, the skeleton, chemotherapy, and even new molecular targets that are still under investigation. Today, the best sequence of treatment for each patient has not been established yet. Objective: The objective of this work is to review the current scene of treatment in castrate resistant prostate cancer, as well as the latest developments and strategies to choose the best sequence in each patient. Material and methods: A literature review was performed through Medline Database (Pubmed) using as key words: "Castrate Resistant Prostate Cancer", "Sequencing", "Biomarkers", "Systemic Therapy". We also reviewed ASCO GU 2017 abstracts. RESULTS: Since Docetaxel was approved in 2004, which increased overall survival by about 2 months in patients with Metastatic Castration Resistant Prostate Cancer, in recent years a large number of therapies have been approved, demonstrating an increase in overall survival after several phase III clinical trials: Cabacitaxel, Abiraterone, Enzalutamide, Sipuleucel-T, Denosumab, Radium 223. And more recently, some investigations about new targeted therapies directed to the androgen receptor, with greater affinity than enzalutamide, or more accurate inhibitors of CYP 17 enzyme than abiraterone, as well as, agents as monoclonal antibodies (anti PD1), vaccines, poly adenosine diphosphate-ribose polymerase inhibitors, are coming to the light. In the future, these outcomes could tune up the treatment sequencing, through the study of predictive biomarkers that will indicate the right target of each therapy. Conclusions: In the near future, outcomes of different clinical trials that are studying new molecules, will allow us to apply the sequencing of different therapies based on biomarkers present in blood (circulating tumor cells) or in specimen biopsies, achieving an increase in overall survival and improving quality of life of patients in the advanced stage of the disease, however the best choice of sequence is unknown at this moment

Porcine Y-chromosome variation is consistent with the occurrence of paternal gene flow from non-Asian to Asian populations.

Pigs (Sus scrofa) originated in Southeast Asia and expanded to Europe and North Africa approximately 1 MYA. Analyses of porcine Y-chromosome variation have shown the existence of two main haplogroups that are highly divergent, a result that is consistent with previous mitochondrial and autosomal data showing that the Asian and non-Asian pig populations remained geographically isolated until recently. Paradoxically, one of these Y-chromosome haplogroups is extensively shared by pigs and wild boars from Asia and Europe, an observation that is difficult to reconcile with a scenario of prolonged geographic isolation. To shed light on this issue, we genotyped 33 Y-linked SNPs and one indel in a worldwide sample of pigs and wild boars and sequenced a total of 9903 nucleotide sites from seven loci distributed along the Y-chromosome. Notably, the nucleotide diversity per site at the Y-linked loci (0.0015 in Asian pigs) displayed the same order of magnitude as that described for autosomal loci (~0.0023), a finding compatible with a process of sustained and intense isolation. We performed an approximate Bayesian computation analysis focused on the paternal diversity of wild boars and local pig breeds in which we compared three demographic models: two isolation models (I models) differing in the time of isolation and a model of isolation with recent unidirectional migration (IM model). Our results suggest that the most likely explanation for the extensive sharing of one Y-chromosome haplogroup between non-Asian and Asian populations is a recent and unidirectional (non-Asian > Asian) paternal migration event.

Knockdown of CXCL14 disrupts neurovascular patterning during ocular development.

The C-X-C motif ligand 14 (CXCL14) is a recently discovered chemokine that is highly conserved in vertebrates and expressed in various embryonic and adult tissues. CXCL14 signaling has been implicated to function as an antiangiogenic and anticancer agent in adults. However, its function during development is unknown. We previously identified novel expression of CXCL14 mRNA in various ocular tissues during development. Here, we show that CXCL14 protein is expressed in the anterior eye at a critical time during neurovascular development and in the retina during neurogenesis. We report that RCAS-mediated knockdown of CXCL14 causes severe neural defects in the eye including precocious and excessive innervation of the cornea and iris. Absence of CXCL14 results in the malformation of the neural retina and misprojection of the retinal ganglion neurons. The ocular neural defects may be due to loss of CXCL12 modulation since recombinant CXCL14 diminishes CXCL12-induced axon growth in vitro. Furthermore, we show that knockdown of CXCL14 causes neovascularization of the cornea. Altogether, our results show for the first time that CXCL14 plays a critical role in modulating neurogenesis and inhibiting ectopic vascularization of the cornea during ocular development.

Análisis del embarazo adolescente: miradas cualitativas a los casos de Bucaramanga y Jaén / Analysis of adolescent pregnancy: a qualitative approach at the cases of Bucaramanga and Jaén

Anualmente millones de jóvenes menores de edad quedan embarazadas, exponiéndose a una serie de riesgos biológicos relacionados con la gestación y a diversas consecuencias sociales. Objetivos: Conocer las percepciones y los imaginarios de las adolescentes frente al embarazo a temprana edad, en una región de Andalucía (España) y otra de Colombia. Material y métodos: Se realizó un estudio cualitativo con enfoque fenomenológico. Se entrevistó a 31 mujeres adolescentes en diferentes periodos del embarazo y el posparto. El contacto con las participantes se efectuó a través de servicios públicos sociales o sanitarios de cada uno de los contextos de estudio. Se llevó a cabo un análisis de contenido de las categorías emergentes, mediante un proceso cíclico, reflexivo, flexible y metódico, adaptado del procedimiento propuesto por Miles y Huberman. Resultados: Se han identificado elementos en común en la maternidad adolescente, independientemente del contexto en el que se desarrolla. Las percepciones y creencias sobre un embarazo adolescente están poco definidas y desarrolladas. Algunas jóvenes interpretaron su embarazo y maternidad en términos positivos y de cumplimiento de un proyecto de vida, aunque las implicaciones negativas fueron más argumentadas. Conclusiones: Los resultados de este estudio ponen de relieve la complejidad de las creencias y percepciones hacia el embarazo adolescente. Las intervenciones en salud sexual y reproductiva dirigidas a la población adolescente deberían tener en cuenta esta diversidad de sentimientos y conductas (AU)

Every year millions of underage girls become pregnant and are exposed to a series of biological risks associated with pregnancy and a variety of social consequences. Objective: The objective of this study is to know the perceptions and imaginary of the adolescents on the pregnancies at early age, in a region of Andalusia (Spain) and another region of Colombia. Materials and methods: A qualitative study with phenomenological approach was conducted. We interviewed 31 adolescent women in different periods of pregnancy and postpartum. Contact with the participants is carried out through public social services or health of each of the contexts of study. We realized a content analysis of the emergent categories, across a cyclical, reflective, flexible and methodical, adapted process of the procedure proposed by Miles and Huberman. Results: We have identified some elements in common in the adolescent maternity, independently of the context in which it develops. Perceptions and beliefs about a teenage pregnancy are poorly defined and developed. Some young women interpret its pregnancy in positive terms and they think that they fulfill a life project although the negative implications are more argued. Conclusions: Our results highlight the complexity of beliefs and perceptions toward pregnancy in adolescence. Sexual and reproductive health interventions targeting adolescent population should take into account the variety of feelings and behaviors (AU)

Compartmentalized Immune Response in Leishmaniasis: Changing Patterns throughout the Disease.

Visceral leishmaniasis (VL) is characterized by loss of T-cell responsiveness and absence of Leishmania-specific IFN-γ production by peripheral blood mononuclear cells. However, the expressions of IFN-γ and TNF-α are up-regulated in the tissues and plasma of VL patients. There is a paucity of information regarding the cytokine profile expressed by different target tissues in the same individual and the changes it undergoes throughout the course of infection. In this work we evaluated IFN-γ, TNF-α, IL-10, and TGF-ß mRNA expression using real-time RT-PCR in 5 target tissues at 6 months and 16 months post-infection (PI) in a canine experimental model which mimics many aspects of human VL. The spleen and liver of Leishmania infantum experimentally-infected dogs elicited a pro- and anti- inflammatory response and high parasite density at 6 and 16 months PI. The popliteal lymph node, however, showed an up-regulation of IFN-γ cytokin at commencement of the study and was at the chronic phase when the IL-10 and TGF-ß expression appeared. In spite of skin parasite invasion, local cytokine response was absent at 6 months PI. Parasite growth and onset of clinical disease both correlated with dermal up-regulation of all the studied cytokines. Our VL model suggests that central target organs, such as the spleen and liver, present a mixed cytokine immune response early on infection. In contrast, an anti-inflammatory/regulatory immune response in peripheral tissues is activated in the later chronic-patent stages of the disease.

PlexinD1 is required for proper patterning of the periocular vascular network and for the establishment of corneal avascularity during avian ocular development.

The anterior eye is comprised of an avascular cornea surrounded by a dense periocular vascular network and therefore serves as an excellent model for angiogenesis. Although signaling through PlexinD1 underlies various vascular patterning events during embryonic development, its role during the formation of the periocular vascular network is yet to be determined. Our recent study showed that PlexinD1 mRNA is expressed by periocular angioblasts and blood vessels during ocular vasculogenesis in patterns that suggest its involvement with Sema3 ligands that are concurrently expressed in the anterior eye. In this study, we used in vivo knockdown experiments to determine the role of PlexinD1 during vascular patterning in the anterior eye of the developing avian embryos. Knockdown of PlexinD1 in the anterior eye caused mispatterning of the vascular network in the presumptive iris, which was accompanied by lose of vascular integrity and profuse hemorrhaging in the anterior chamber. We also observed ectopic vascularization of the cornea in PlexinD1 knockdown eyes, which coincided with the formation of the limbal vasculature in controls. Finally we show that Sema3E and Sema3C transcripts are expressed in ocular tissue that is devoid of vasculature. These results indicate that PlexinD1 plays a critical role during vascular patterning in the iris and limbus, and is essential for the establishment of corneal avascularity during development. We conclude that PlexinD1 is involved in vascular response to antiangiogenic Sema3 signaling that guides the formation of the iris and limbal blood vessels by inhibiting VEGF signaling.

Angiosarcoma primario y secundario de mama: estudio de 8 casos con evidencia de origen linfático en los angiosarcomas posradioterapia / Primary and secondary angiosarcoma of the breast. A study of 8 cases with evidence of a lymphatic origin in post-radiation angiosarcomas

Objetivo. Analizar las diferencias entre los angiosarcomas primarios y posradioterapia de mama. Pacientes y métodos. Revisamos retrospectivamente angiosarcomas de mama entre los años 2000 y 2010. Realizamos un estudio clinicopatológico e inmunohistoquímico con CKAE1/AE3, CD31, CD34, Ki-67, D2-40. Resultados. Se incluyeron 8 mujeres, 4 con angiosarcomas primarios y 4 secundarios. La edad media era de 66 años en los primarios y de 74 en los secundarios. El periodo de latencia medio posradioterapia en los angiosarcomas secundarios fue de 118 meses. Tres tumores secundarios afectaban la piel, 3 angiosarcomas primarios eran intraparenquimatosos y los 2 restantes fueron mixtos. Todos los casos fueron CD31+/CD34+/CKAE1/AE3−. Los angiosarcomas secundarios expresaron el marcador linfático D2-40, mientras que los primarios eran D2-40−. Conclusiones. Los angiosarcomas secundarios expresan D2-40, mientras que los primarios son negativos para este marcador. Ello evidencia un origen vascular linfático para los angiosarcomas posradioterapia (AU)

Objective. To analyze differences between primary and radiation-associated secondary breast angiosarcomas. Patients and methods. We retrospectively reviewed all cases of angiosarcoma diagnosed at our hospital between 2000 and 2010. We analyzed the clinical and pathological features. In the immunohistochemical study, we assessed expression of CKAE1/AE3, CD31, CD34, Ki-67 and D2-40. Results. There were 8 women, 4 with primary angiosarcoma and 4 with secondary angiosarcoma. The mean age at presentation was 66 years for primary tumors and 74 years for secondary angiosarcomas. The mean latency period for radiation-associated angiosarcomas was 118 months. Three secondary tumors involved the skin, 3 primary angiosarcomas were intramammary and the remaining 2 were mixed. All tumors were CD31+/CD34+/CKAE1/AE3−. The secondary angiosarcomas also expressed D2-40, while the primary tumors were negative for this lymphatic marker. Conclusions. Secondary angiosarcomas express D2-40, while primary angiosarcomas are negative for this lymphatic marker. This finding suggests a lymphatic origin for post-radiation angiosarcomas (AU)

Sema3A maintains corneal avascularity during development by inhibiting Vegf induced angioblast migration.

Corneal avascularity is important for optical clarity and normal vision. However, the molecular mechanisms that prevent angioblast migration and vascularization of the developing cornea are not clear. Previously we showed that periocular angioblasts and forming ocular blood vessels avoid the presumptive cornea despite dynamic ingression of neural crest cells. In the current study, we investigate the role of Semaphorin3A (Sema3A), a cell guidance chemorepellent, on angioblast migration and corneal avascularity during development. We show that Sema3A, Vegf, and Nrp1 are expressed in the anterior eye during cornea development. Sema3A mRNA transcripts are expressed at significantly higher levels than Vegf in the lens that is positioned adjacent to the presumptive cornea. Blockade of Sema3A signaling via lens removal or injection of a synthetic Sema3A inhibitor causes ectopic migration of angioblasts into the cornea and results in its subsequent vascularization. In addition, using bead implantation, we demonstrate that exogenous Sema3A protein inhibits Vegf-induced vascularization of the cornea. In agreement with these findings, loss of Sema/Nrp1 signaling in Nrp1(Sema-) mutant mice results in ectopic angioblasts and vascularization of the embryonic mouse corneas. Altogether, our results reveal Sema3A signaling as an important cue during the establishment of corneal avascularity in both chick and mouse embryos. Our study introduces cornea development as a new model for studying the mechanisms involved in vascular patterning during embryogenesis and it also provides new insights into therapeutic potential for Sema3A in neovascular diseases.

Expression of CXCL12 and CXCL14 during eye development in chick and mouse.

Vertebrate eye development is a complex multistep process coordinated by signals from the lens, optic cup and periocular mesenchyme. Although chemokines are increasingly being recognized as key players in cell migration, proliferation, and differentiation during embryonic development, their potential role during eye development has not been examined. In this study, we demonstrate by section in situ hybridization that CXCL12 and CXCL14 are expressed during ocular development. CXCL12 is expressed in the periocular mesenchyme, ocular blood vessels, retina, and eyelid mesenchyme, and its expression pattern is conserved between chick and mouse in most tissues. Expression of CXCL14 is localized in the ocular ectoderm, limbal epithelium, scleral papillae, eyelid mesenchyme, corneal keratocytes, hair follicles, and retina, and it was only conserved in the upper eyelid ectoderm of chick and mouse. The unique and non-overlapping patterns of CXCL12 and CXCL14 expression in ocular tissues suggest that these two chemokines may interact and have important functions in cell proliferation, differentiation and migration during eye development.

Availability and costs of single cigarettes in Guatemala.

INTRODUCTION: Single-cigarette sales have been associated with increased cigarette accessibility to less educated, lower-income populations, and minors; lower immediate cost, and increased smoking cues. Since 1997, Guatemalan Law bans the sale of single cigarettes and packs with fewer than 20 cigarettes. In 2005, Guatemala ratified the World Health Organization Framework Convention on Tobacco Control (WHO FCTC); it is therefore obliged to "prohibit sale of cigarettes individually or in small packets." METHODS: Blocks were numbered and randomly selected in Guatemala City and 3 neighboring towns. All stores in each block were surveyed. Single-cigarette and fewer than 20-cigarette pack sales were assessed by observation and purchase attempts. Cigarette brands and manufacturers (Philip Morris, PM or British American Tobacco, BAT) were also recorded. Percentages and means were used to describe data. Analyses were done using STATA 11.0. RESULTS: Of 398 stores and street vendors surveyed, 75.6% (301) sold cigarettes. Of these, 91% (275) sold single cigarettes and none sold fewer than 20-cigarette packs. Only informal economic sectors sold singles. There was no difference on sales between Guatemala City and neighboring towns and by store type. Buying 20 single cigarettes was US$ 0.83 more expensive than buying a 20-cigarette pack. The most prevalent brands were Rubios (PM), Marlboro (PM), Payasos (BAT), and After Hours (BAT). CONCLUSIONS: Single-cigarettes sales are highly prevalent among informal economic sectors in Guatemala City and its neighboring towns. Our data should prove useful to advocate for FCTC Article 16 enforcement in Guatemala.

Performance of commercially available serological diagnostic tests to detect Leishmania infantum infection on experimentally infected dogs.

Leishmania infantum (syn. Leishmania chagasi) is the etiological agent of a widespread serious zoonotic disease that affects both humans and dogs. Prevalence and incidence of the canine infection are important parameters to determine the risk and the ways to control this reemergent zoonosis. Unfortunately, there is not a gold standard test for Leishmania infection. Our aim was to assess the operative validity of commercial tests used to detect antibodies to Leishmania in serum samples from experimental infections. Three ELISA tests (LEISCAN(®) Leishmania ELISA Test, INGEZIM(®) LEISHMANIA, and INGEZIM(®) LEISHMANIA VET), three immunochromatographic tests (INGEZIM(®) LEISHMACROM, SNAP(®) Leishmania, and WITNESS(®) Leishmania), and one IFAT were evaluated. LEISCAN(®) Leishmania ELISA test achieved the highest sensitivity and accuracy (both 0.98). Specificity was 1 for all tests except for IFAT. All tests but IFAT obtained a positive predictive value of 1, while the maximum negative predictive value was achieved by LEISCAN(®) Leishmania ELISA Test (0.93). The best positive likelihood ratio was obtained by INGEZIM(®) LEISHMANIA VET (30.26), while the best negative likelihood ratio was obtained by LEISCAN(®) Leishmania ELISA Test (0.02). The highest diagnostic odds ratio was achieved by LEISCAN(®) Leishmania ELISA Test (729.00). The largest area under the ROC curve was obtained by LEISCAN(®) Leishmania ELISA Test (0.981). Quantitative ELISA based tests performmed better than qualitative tests ("Rapid Tests"), and the test best suited to detect Leishmania in infected dogs and to provide clinically useful information was LEISCAN(®) Leishmania ELISA Test. This and other results point also to the need of revising the status of IFAT as a gold standard for the diagnosis of leishmaniasis.

Efficacy and safety of concurrent trastuzumab plus weekly paclitaxel-FEC as primary therapy for HER2-positive breast cancer in everyday clinical practice.

One of the most efficacious primary therapies in HER2-positive breast cancer was published by the M.D. Anderson group in 2005. This randomized trial evaluated the addition of trastuzumab to a taxane-anthracycline based chemotherapy. Despite largely significant differences in pathological complete response (pCR) in the trastuzumab group (65 vs. 26 %) this regimen did not become a common standard due to toxicity concerns and its premature closure with a small sample size. In order to evaluate the efficacy and safety of this regimen in an off-trial setting we conducted a prospectively monitorized series of consecutive patients with early or locally advanced Her-2 positive breast cancer following the same treatment strategy. Stage II-IIIC HER2-positive breast cancer patients, including inflammatory disease, were treated with weekly-trastuzumab for 24 weeks administered concurrently with all primary chemotherapy containing paclitaxel (80 mg/m(2)) for 12 weeks and 4 cycles of FEC-75 (fluorouracil 500 mg/m(2), epirubicine 75 mg/m(2), and cyclophosphamide 500 mg/m(2)) followed by surgery. The objectives were efficacy, in terms of pCR in both the breast and lymph nodes, and safety, with close cardiac monitoring during and after treatment. From August 2004 to February 2009, 83 patients were included. Most patients (73.5 %) had node involvement and 13.2 % had inflammatory disease. Fifty-one patients (61.4 %) achieved a pCR in breast and axilla (95 % CI 50-72 %). HR-negative tumors were associated with higher pCR rate than HR-positive tumors (77 vs. 48 %, P = 0.006). At a median follow-up of 50.2 months no patient developed symptomatic cardiac failure, and 9 patients (10.8 %) presented a transient asymptomatic decrease in left ventricular ejection fraction. Primary therapy with concurrent trastuzumab plus paclitaxel-FEC for HER2-positive breast cancer in everyday practice is highly effective and safe confirming the results observed in a randomized trial stopped prematurely.

Possible important pair of acidic residues in vesicular acetylcholine transporter.

Invariant E309 is in contact with critical and invariant D398 in a three-dimensional homology model of vesicular acetylcholine transporter (VAChT, TC 2.A.1.2.13) [Vardy, E., et al. (2004) Protein Sci. 13, 1832-1840]. In the work reported here, E309 and D398 in human VAChT were mutated singly and together to test their functions, assign pK values to them, and determine whether the residues are close to each other in three-dimensional space. Mutants were stably expressed in the PC12(A123.7) cell line, and transport and binding properties were characterized at different pH values using radiolabeled ligands and filtration assays. Contrary to a prior conclusion, the results demonstrate that most D398 mutants do not bind the allosteric inhibitor vesamicol even weakly. Earlier work showed that most D398 mutants do not transport ACh. D398 therefore probably is the residue that must deprotonate with a pK of 6.5 for binding of vesamicol and with a pK of approximately 5.9 for transport of ACh. Because E309Q has no effect on VAChT functions at physiological pH, E309 has no apparent critical role. However, radical mutations in E309 cause decreases in ACh and vesamicol affinities and total loss of ACh transport. Unlike wild-type VAChT, which exhibits a peak of [(3)H]vesamicol binding centered at pH 7.4, mutants E309Q, E309D, E309A, and E309K all exhibit peaks of binding centered at pH >or=9. The combination of high pH and mutated E309 apparently produces a relaxed (in contrast to tense) conformation of VAChT that binds vesamicol exceptionally tightly. No compensatory interactions between E309 and D398 in double mutants were discovered. Proof of a close spatial relationship between E309 and D398 was not found. Nevertheless, the data are more consistent with the homology model than an alternative hydropathy model of VAChT that likely locates E309 far from D398 and the ACh binding site in three-dimensional space. Also, a probable network of interactions involving E309 and an unknown residue having a pK of 10 has been revealed.