Estrogen receptor subtype beta2 is involved in neuromast development in zebrafish (Danio rerio) larvae

Estrogens are known to play a role in both reproductive and non-reproductive functions in mammals. Estrogens and their receptors are involved in the development of the central nervous system (brain development, neuronal survival and differentiation) as well as in the development of the peripheral nervous system (sensory-motor behaviors). In order to decipher possible functions of estrogens in early development of the zebrafish sensory system, we investigated the role of estrogen receptor beta(2) (ERbeta(2)) by using a morpholino (MO) approach blocking erbeta(2) RNA translation. We further investigated the development of lateral line organs by cell-specific labeling, which revealed a disrupted development of neuromasts in morphants. The supporting cells developed and migrated normally. Sensory hair cells, however, were absent in morphants' neuromasts. Microarray analysis and subsequent in situ hybridizations indicated an aberrant activation of the Notch signaling pathway in ERbeta(2) morphants. We conclude that signaling via ERbeta(2) is essential for hair cell development and may involve an interaction with the Notch signaling pathway during cell fate decision in the neuromast maturation process.

Zebrafish (Danio rerio) neuromast: promising biological endpoint linking developmental and toxicological studies

Aquatic toxicology is facing the challenge to assess the impact of complex mixtures of compounds on diverse biological endpoints. So far, ecotoxicology focuses mainly on apical endpoints such as growth, lethality and reproduction, but does not consider sublethal toxic effects that may indirectly cause ecological effects. One such sublethal effect is toxicant-induced impairment of neurosensory functions which will affect important behavioural traits of exposed organisms. Here, we critically review the mechanosensory lateral line (LL) system of zebrafish as a model to screen for chemical effects on neurosensory function of fish in particular and vertebrates in general. The LL system consists of so-called neuromasts, composed of centrally located sensory hair cells, and surrounding supporting cells. The function of neuromasts is the detection of water movements that is essential for the fish's ability to detect prey, to escape predator, to socially interact or to show rheotactic behaviour. Recent advances in the study of these organs provided researchers with a broad area of molecular tools for easy and rapid detection of neuromasts dysfunction and/or disturbed development. Further, genes involved in neuromasts differentiation have been identified using auditory/mechanosensory mutants and morphants. A number of environmental toxicants including metals and pharmaceuticals have been shown to affect neuromasts development and/or function. The use of the LL organ for toxicological studies offers the advantage to integrate the available profound knowledge on developmental biology of the neuromasts with the study of chemical toxicity. This combination may provide a powerful tool in environmental risk assessment.

Long-term regulation of neuronal high-affinity glutamate and glutamine uptake in Aplysia.

An increase in transmitter release accompanying long-term sensitization and facilitation occurs at the glutamatergic sensorimotor synapse of Aplysia. We report that a long-term increase in neuronal Glu uptake also accompanies long-term sensitization. Synaptosomes from pleural-pedal ganglia exhibited sodium-dependent, high-affinity Glu transport. Different treatments that induce long-term enhancement of the siphon-withdrawal reflex, or long-term synaptic facilitation increased Glu uptake. Moreover, 5-hydroxytryptamine, a treatment that induces long-term facilitation, also produced a long-term increase in Glu uptake in cultures of sensory neurons. The mechanism for the increase in uptake is an increase in the V(max) of transport. The long-term increase in Glu uptake appeared to be dependent on mRNA and protein synthesis, and transport through the Golgi, because 5,6-dichlorobenzimidazole riboside, emetine, and brefeldin A inhibited the increase in Glu uptake. Also, injection of emetine and 5,6-dichlorobenzimidazole into Aplysia prevented long-term sensitization. Synthesis of Glu itself may be regulated during long-term sensitization because the same treatments that produced an increase in Glu uptake also produced a parallel increase in Gln uptake. These results suggest that coordinated regulation of a number of different processes may be required to establish or maintain long-term synaptic facilitation.

(Doctor perplexorum) oder: theologisch-philosophische Erörterungen über die Übereinstimmung der mosaischen und rabbinischen Religionsquellen mit der Philosophie

von Rabbi Moses Maimonides. Aus dem Hebr. des Ebn Thybbon ins Dt. übers., und kommentiert von R. I. Fürstenthal

The effects of proteasome inhibition on angiogenesis and autophagy in human prostate cancer cells

The proteasome degrades approximately 80% of intracellular proteins to maintain homeostasis. Proteasome inhibition is a validated therapeutic strategy, and currently, proteasome inhibitor bortezomib is FDA approved for the treatment of MM and MCL. Specific pathways affected by proteasome inhibition have been identified, but mechanisms of the anti-tumor effects of proteasome inhibition are not fully characterized and cancer cells display marked heterogeneity in terms of their sensitivity to proteasome inhibitor induced cell death. ^ The antitumor effects of proteasome inhibition involve suppression of tumor angiogenesis and vascular endothelial growth factor (VEGF) expression, but the mechanisms involved have not been clarified. In this dissertation I investigated the mechanisms underlying the effects of two proteasome inhibitors, bortezomib and NPI-0052, on VEGF expression in human prostate cancer cells. I found that proteasome inhibitors selectively downregulated hypoxia inducible factor 1alpha (HIF-1α) protein and its transcriptional activity to inhibit VEGF expression. Mechanistic studies demonstrated that proteasome inhibitors mediate the induction of the unfolded protein response (UPR) and that downregulation of HIF-1α is caused by eukaryotic translation initiation factor 2α (eIF2α) phosphorylation and translation repression. Importantly, I showed that proteasome inhibitors activated the UPR in some cells but not in others. My observation may have implications for the design of combination regimens that are based on exploiting proteasome inhibitor-induced ER stress.^ Although proteasome inhibitors have shown modest activity on prostate cancer, there is general consensus that no single agent is likely to have significant activity in prostate cancer. In the second part of this dissertation I attempted to exploit the effects of proteasome inhibition on the UPR to design a combination therapy that would enhance cancer cell death. Autophagy is a lysosome dependent degradation pathway that functions to eliminate long-lived protein and subcellular structures. Targeting autophagy has been shown to inhibit tumors in preclinical studies. I found that inhibition of autophagy with chloroquine or 3-methyladenine enhanced proteasome inhibitor induced cell death and the effects were associated with increased intracellular stress as marked by aggresome formation. Multiple cancers appear to be resistant to proteasome inhibition treatment alone. The implications of synergy for the combined inhibition of autophagy and the proteasome would likely apply to other cancers aside from prostate cancer. ^

For health's sake: Cervical and breast cancer screening

African-Americans make up twelve percent of the United States population, yet they experience morbidity and mortality at a rate that, in some cases, is disproportionate to their numbers. There are numerous health areas, including cancer, in which disparities exist. There are also numerous reasons which have been suggested to explain the high rates of cancer morbidity and mortality experienced by African-Americans. Among the reasons given to explain these differences are lack of knowledge and lack of access to medical care (1). This study sought to increase the knowledge, attitudes, and behavioral intentions of African-American women attending a Baptist church in Houston with regard to cervical cancer, breast cancer, Pap smear, and mammography. It was hypothesized that a church-based cancer education program would produce the desired change in knowledge, attitudes, and behavioral intentions.^ The quasi-experimental design of the study was untreated control group with pretest and posttest and untreated control group with posttest only. Female members of Mount Ararat Baptist Church took part in an eight-week, cancer education program based on social cognitive theory. Baseline data were collected before the start of the program at Mount Ararat and at Solid Rock Baptist Church, control group one. At the end of the program, the follow-up survey was administered at the program church, control church one, and in a third church, Damascus Missionary Baptist Church, which served as the posttest only group. The data were analyzed by Fisher's exact and paired t-test to determine if the program supported the project's hypotheses.^ Results of data analyses supported the major study hypotheses, the exception being behavioral intention to have Pap smear performed. Although the program appeared to have generally influenced changes in the desired direction, the results are limited due to the quasi-experimental design and small sample size. Longer term studies with larger sample sizes are needed to more fully develop and evaluate programs which impact the health of African-Americans. ^

Transcriptional regulation of the {\it neu\/} oncogene and its negative regulation by the c-{\it myc\/} oncogene

The first part of my research involved the characterization of the neu gene promoter. I subcloned a 2.2-kb sequence located upstream to the extreme 5$\sp\prime$ end of the neu gene, in front of the bacterial reporter gene, chloramphenicol acetyltransferase (CAT). Transfection of this construct into different cell lines and subsequent CAT assays demonstrated that this 2.2-kb fragment was functional as a promoter. A series of deletion constructs was engineered to study the contribution of different fragments to transcription. Subcloning of individual fragments was followed by a cotransfection competition experiment, which demonstrated the involvement of protein factors interacting with the promoter. A gel retardation assay was also performed to show the physical binding of protein factors to the promoter. The combined results suggested that both positively and negatively acting protein factors are involved in interacting with different regions of the promoter, contributing to the overall transcription activity. My findings provide an insight into the regulation of neu gene expression, which in turn provides the tools to understand the molecular mechanisms of overexpression of the neu gene in some breast cancer and ovarian cancer cell lines.^ In the second part of my research, I discovered that another oncogene, c-myc, was able to reverse the transformed morphology that was induced by the neu oncogene. Utilizing the promoter constructs that I made, I was able to show that the c-myc oncogene has a negative regulatory effect on the expression of the neu oncogene. Further studies suggested that c-myc is able to lower the effective concentration of a positive factor(s) that interact with a 139-bp fragment of the neu gene promoter. These findings may provide a direct evidence of the long suspected role of the c-myc gene in transcriptional regulation. The neu gene may very well be the first identified mammalian target gene that is regulated by the c-myc oncogene. Since c-myc is known to be stimulated by various mitogenic signals and the neu gene is likely to be a growth factor receptor, it is possible that c-myc, when stimulated by the signal transduction pathway of the neu gene, would function as a negative feedback regulator on the neu gene receptor. (Abstract shortened with permission of author.) ^

Contaminación por fosfatos en el oasis bajo riego del río Mendoza

El oasis bajo riego del río Mendoza, en la provincia argentina del mismo nombre -al igual que casi todas las ciudades en la actualidadpresenta problemas de avance de la urbanización sobre las tierras agrícolas, multiplicidad de usuarios y disminución de la disponibilidad del recurso hídrico, tanto en cantidad como en calidad. Si bien se destinan esfuerzos e inversiones tendientes a asegurar la disponibilidad de agua (mejora de eficiencias, ahorro de agua) no pasa lo mismo en relación con la preservación de su calidad. La agricultura mendocina resulta víctima de la contaminación producida por la urbanización y la industria a través del vuelco (puntual y/o difuso) de sus efluentes a la red de riego. Estudios realizados en el Oasis Norte de Mendoza pusieron de manifiesto la existencia de altos niveles de contaminación fosfatada en las aguas del río Mendoza. El presente trabajo tuvo como objetivo evaluar la evolución espacio-temporal y detectar las fuentes de esta contaminación. Los resultados del diagnóstico basado en una serie de muestreos realizados en 2003 - 2009 ponen de relieve la existencia de una moderada contaminación por fosfatos en las aguas del río Mendoza que riegan el Oasis Norte provincial. Asimismo, se detectaron niveles considerablemente altos de fosfatos en tres sitios específicos del oasis: 1. la superficie regadía servida por los canales Cacique Guaymallén y Jocolí -se observa un incremento de seis veces el contenido de fosfatos del agua: de 0,2 mg L-1 (R I) a 1,2 mg L-1 (C II)-; en este último sitio sólo se riega un pequeño sector que se aproxima a las 7.300 ha; 2. la superficie regada por el Colector Pescara aguas abajo del punto D VIII (1.250 ha), en la que los valores medios arrojaron un contenido diecisiete veces mayor (8,5 mg L-1 ) que los del sitio D I (0,49 mg L-1 ) que recibe desagües agrícolas y urbano pluviales; 3. la zona del Bajo río Mendoza (en esta zona se registró un aumento de dieciséis veces más fosfatos entre la parte media y la cola del sistema, con valores medios de 0,2 mg L-1 en el sitio R II y de 3,25 mg L-1 en R III).

Memoria de la segunda revision del monte "Poyo de Santo Domingo", de la tercera revision del monte "Cerros del Pozo" y de la primera revision del monte "Cerros de Hinojares"

EL día 24 le Junio de 1.941 me fue entregado en la secretaría de la Escuela Especial de Ingenieros de Montes el oficio que a continuación t r a n s c r i b o; rt Ministerio de cultura -Dirección General de Montes Caza y Pesca aluvial- Con esta fecha digo al Sr. Ingeniero Jefe del Distrito Forestal de Jaén lo siguiente:- Con el fin de suplir en lo posible la escasez de personal técnico afecto a ese servicio, para llevar a cabo el estudio y redacción de proyectos necesarios, este Centro ha dispuesto de acuerdo con la Dirección de la Escuela Especial de Ingenieros de Montes y teniendo en cuenta las manifestaciones de la Jefatura, designar a los alumnos del último curso Don Vicente Reus Cid, Don Juan* José Casado Bracho y Don Rafael "Fernández Huerta para que trabajen realizando los estudios que se l e encomienden a las inmediatas órdenes del Ingeniare de esa Dependencia, designado por V. S., que será quien lleve la dirección y responsabilidad del trabajo para presentarlo con una ante esa Jefatura que lo remitirá, con el regí amen t al informe a la dirección General rmine.- Ba su día se fijara por la Superioridad el parecer de la Dirección dé la Escuela y el de V.3. la parte de las indemnizaciones correspondiente a la tarifa reglamentaria que deberá percibir cada alumno que colabore con el Ingeniero -designado .- Las revisiones deberán estar terminadas para fin del año corriente. Lo que traslado para su conocimiento y efectos.