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Fundamento y objetivo Evaluar la mortalidad tras el alta de la Unidad de Cuidados Intensivos �post-UCI� (hospitalaria y seguimiento a un ano) y los factores asociados. Pacientes y metodo Diseno de cohortes en enfermos medicoquirurgicos con sindrome de disfuncion multiorganica (SDMO) durante sus primeras 24h de ingreso. Se registraron antecedentes personales, situacion basal, datos generales de ingreso en UCI, estancia hospitalaria y datos de supervivencia mediante contacto telefonico al ano. Se evaluo la mortalidad en cualquier momento del seguimiento, relizando una regresion de Cox para valorar factores de mortalidad. Resultados Se reclutaron 545 pacientes. En el total del periodo de estudio fallecieron 256 pacientes (52,9%); de los ingresados en UCI fallecieron el 29,5%, mientras que de los 384 enfermos que pasaron a planta fallecio el 14,8%. De los 327 enfermos dados de alta hospitalaria se ha contactado con el 81,3% (266 pacientes), de los que han fallecido el 14,3%. Los factores relacionados con la muerte hospitalaria fueron la edad (odds ratio [OR] 1,04; intervalo de confianza del 95% [IC 95%] 1,02-1,06; p

Resumen Objetivos Estudiar la incidencia del «trastorno por duelo prolongado» desde un ano despues del fallecimiento de un familiar ingresado en la Unidad de Cuidados Intensivos (UCI). Material y metodos Estudio de seguimiento longitudinal con corte transversal en una UCI polivalente de un hospital de referencia. Se evaluo a los allegados alrededor de un ano despues del fallecimiento, usando como herramientas los «criterios de consenso para el diagnostico del trastorno por duelo prolongado» (CCTDP). Se determino la prevalencia entre el primero y segundo anos. Resultados Se incluyeron 151 familiares de fallecidos en la UCI. El seguimiento se realizo 22,1 ± 5,3 meses tras el fallecimiento. Once familiares (10,3%) cumplieron los CCTDP. De todos los allegados en duelo, aquellos identificados con trastorno por duelo prolongado son los que con mas frecuencia precisaron apoyo psicologico/psiquiatrico. Conclusiones En una muestra de allegados de pacientes fallecidos tras un ingreso en la UCI, una minoria significativa cumplio los CCTDP al cabo de 1-2 anos tras el fallecimiento. Hay que tener en consideracion este hecho que es frecuentemente pasado por alto, y que podria precisar algun tipo de intervencion psicologica.

Resumen Objetivo Valorar si el gradiente alveolo-arterial de oxigeno (Grad[A-a]O 2 ) ayuda a confirmar la influencia de la PEEP en la PaFi (PaO 2 /FiO 2 ). Diseno Estudio observacional; usamos una regresion lineal para realizar un estudio multivariable y mejorar la formula de la PaFi teniendo en cuenta la PEEP. Ambito Hospital terciario. Pacientes Todos los pacientes de cuidados intensivos, con o sin dano pulmonar. Variables de interes principal Registramos los antecedentes personales, juicio clinico, datos durante ingreso en UCI, puntuaciones de severidad en el primer dia y durante evolucion; dos variables calculadas: PaFi and Grad(A-a)O 2 . Resultados Un total de 956 patientes incluidos; 63,9% hombres; edad mediana 68 anos. Primer dia, 31,8% no tienen ventilacion mecanica (VM), 13,1% tienen VM no invasiva y 55,1% VM invasiva. PaFi: 32,9% 0-200, 32,2% 201-300, y 34,8% >300. PEEP: 0-5 69,8%, 6-10 27,5% y >10 2,6%. Observamos una correlacion (Pearson) entre el Grad(A-a)O 2 y la PaFi de -0,84 (p 2 ) de 0,62 sin PEEP y 0,72 con PEEP. Cambiamos la formula de la PaFi, denominandola PaFip (PaFi mas PEEP): Ln (PaFi/[PEEP + 12]). El indice de correlacion entre PaFip y Grad(A-a)O 2 : -0,9 (p 2 ) y utilizamos PaFip en vez de la PaFi. Solo la PaFi permanece en el modelo, y es discretamente complementada por APACHE IV; R 2 0,8. Conclusiones Anadiendo la PEEP a la PaFi, creamos una variable (PaFip) que mejora el modelo, demostrando mayor capacidad de bondad de ajuste.

Physiotherapists are exposed to various occupational stressors, which, if persistent, can lead to the Burnout syndrome, psychosocial phenomenon, consisting of three dimensions: Emotional Exhaustion, Depersonalization and Low Professional Accomplishment. 0bjective: To assess the Burnout syndrome and its association with variables socio-demographic, academic, psychosocial and discomfort/pain.Method: In the search were used MBI-SS (Maslach Burnout Inventory - Student Survey), the scale of Corlet and a questionnaire to identification the demographic variables, academic and psychosocial. The application of instruments was held in places of probation. Results identified average index of Emotional Exhaustion, low Descrença and high rate of effectiveness training, indicating a pre provision for the development of the syndrome. There was also association between the size of Burnout and variable academic, psychosocial and pain, where they had direct relationship with the syndrome. Conclusion: Results point to the need for intervention by the end of current students in order to prevent the Burnout syndrome at work. Los profesionales de la salud están expuestos a diversas manifestaciones de estrés, que, si se mantienen, pueden dar lugar a Síndrome de Burnout, fenómeno psicosocial que consta de tres dimensiones: agotamiento emocional, despersonalización y bajos logros profesionales. El estudio tuvo como objetivo evaluar el Síndrome de Burnout y su asociación con variables sociodemográficas, académicos, psicosociales y malestar/dolor en 101 estudiantes de fisioterapia. Como los motores de búsqueda se utilizaron los SS-MBI (Maslach Burnout Inventory - Student Survey), la escala de Corleto y un cuestionario para identificar las variables demográficas, académicas y psicosociales.Los resultados identificaron tasa media de cansancio emocional, bajo nivel de incredulidad y de alto índice de eficiencia de la ocupación, indicando la posibilidad del riesgo de desarrollar el síndrome. También hay correlación entre el tamaño de Burnout y variables académicas, psicosociales y del dolor, en el que se presentó una relación directa con el síndrome. Los resultados sugieren la necesidad de una intervención al final del curso para los estudiantes, con el fin de evitar el síndrome de Burnout en el ejercicio de capacitación. Fisioterapeutas estão expostos a diversos estressores ocupacionais, que, se persistentes, podem levar à Síndrome de Burnout, fenómeno psicossocial, constituído por três dimensões: Exaustão Emocional, Despersonalização e Baixa Realização Profissional. 0 estudo buscou avaliar a Síndrome de Burnout e sua associação com variáveis sóciodemográficas, acadêmicas, psicossociais e desconforto/dor em 101 estudantes de fisioterapia. Como instrumentos de pesquisa foram utilizados o MBI-SS (Maslach Burnout Inventory - Student Survey), a Escala de Corlet e um questionário para identificação das variáveis demográficas, acadêmicas e psicossociais. Resultados identificaram médio índice de Exaustão Emocional, baixo índice de Descrença e Alto índice de Eficácia profissional, indicando possibilidade de risco de desenvolvimento da síndrome. Verificou-se também associação entre as dimensões de Burnout e variáveis acadêmicas, psicossociais e desconforto/dor, onde estas apresentaram relação direta com a síndrome. Conclusão: Os resultados apontam para a necessidade de intervenção em alunos de final de curso no sentido de prevenir a Síndrome de Burnout no exercício profissional. Palavras chaves: Síndrome de Burnout; estagiários, fisioterapia.

Uterine secretions have a dominant impact on the environment in which embryo development takes place. The uterine serpins (SERPINA14, previously known as UTMP) are found most abundantly during pregnancy in the uterus of ruminants. Although progesterone is currently assumed to be the major regulator of SERPINA14 expression, our recent study of transcriptome changes in bovine endometrium during the estrous cycle unexpectedly detected a marked upregulation of SERPINA14 mRNA levels at estrus. The present study describes the full-length mRNA sequence, genomic organization, and putative promoter elements of the SERPINA14 gene. The SERPINA14 mRNA abundance was quantified by real-time RT-PCR in intercaruncular endometrium at several time points during the estrous cycle and early pregnancy. Highest levels were found at estrus, followed by a dramatic decrease and a moderate expression during the luteal phase. Transcript levels were higher in pregnant endometrium compared with controls at Day 18. At estrus, immunoreactive protein was localized in deep glandular epithelium, and Western blotting concomitantly showed the 52-kDa form in uterine flushings. SERPINA14 mRNA was significantly upregulated in glandular endometrial cells in vitro after stimulation with estradiol-17beta and progesterone, but not after interferon-tau treatment. Our results clearly demonstrate that SERPINA14 appears distinctly in bovine endometrium during the estrus phase. A supporting role toward providing a well-prepared endometrial environment for passing gametes, especially sperm, is assumed.

Prostaglandins (PGs) are important regulators of reproductive processes including early embryonic development. We analyzed the most relevant PG in bovine uteri at different preimplantation pregnancy stages when compared with non-pregnant controls. Additionally, endometrium and trophoblast tissues were examined regarding specific enzymes and receptors involved in PG generation and function. Simmental heifers were artificially inseminated or received seminal plasma only. At days 12, 15, or 18, post-estrus uteri were flushed for PG determination by liquid chromatography–tandem mass spectrometry. Endometrium and trophoblast tissues were sampled for RNA extraction and quantitative real-time PCR analysis. At all days and points of time examined, the concentration of 6-keto PGF1α (stable metabolite of PGI2) was predominant followed by PGF2α>PGE2>PGD2≈TXB2 (stable metabolite of TXA2). At days 15 and 18, PG increased from overall low levels at day 12, with a much more pronounced increase during pregnancy. The PGF2α/PGE2 ratio was not influenced by status. The highest PG concentration was measured at day 15 with 6-keto PGF1α (6.4 ng/ml) followed by PGF2α (1.1 ng/ml) and PGE2 (0.3 ng/ml). Minor changes in endometrial PG biosynthesis enzymes occurred due to pregnancy. Trophoblasts revealed high transcript abundance of general and specific PG synthases contributing to uterine PG. As PGI2 and PGF2α receptors were abundantly expressed by the trophoblast, abundant amounts of PGI2 and PGF2α in the uterine lumen point towards an essential role of PG for the developing embryo. High amounts of PG other than PGE2 in the preimplantation uterus may be essential rather than detrimental for successful reproduction.

BACKGROUND: Immune alterations may be involved in the pathogenesis and progression of endometriosis. Dendritic cells (DCs) are potent antigen presenting cells that are highly involved in the initiation of the immune response. The aim of this study was to investigate DC populations in the eutopic and ectopic endometrium of women with endometriosis compared with controls. METHODS: Hysterectomy samples were obtained from premenopausal women with (n = 33) and without (n = 28) endometriosis. In addition, paired peritoneal endometriotic lesions and uterine curettings were collected from 32 women with endometriosis. Specimen sec- tions were stained immunohistochemically using antibodies for monoclonal mouse antibodies directed against human CD la and CD83, which are specific for immature and mature DCs, respectively. RESULTS: The mean density of endometrial CD I a+ DCs in the basal layer was significantly increased in women with endometriosis com- pared with controls during the proliferative phase only (P = 0.001). There was a highly significant decrease in the density of endometrial CD83+ DCs in women with endometriosis compared with controls in both layers of the endometrium across all phases of the menstrual cycle (P = 0.001). The density of CD la+ DCs was significantly increased in peritoneal endometriotic lesions (P = 0.003) and in the sur- rounding peritoneum (P = 0.00I) compared with paired uterine curettings and peritoneum distant from the lesion. CONCLUSIONS: Both CD la+ and CD83 + DC populations were altered in the eutopic and ectopic endometrium of women with endo- metriosis compared with controls. Alterations in these cells, which play a crucial role in the coordination of the immune response, may be involved in pain generation and the pathogenesis of endometriosis.

Endometriosis is a disease that still presents many puzzles to clinicians and basic research scientists. Until recently, it has been regarded as a condition that arises when normal endometrium adheres to the peritoneal surface and then grows into an ‘inflammatory’ lesion, which adversely influences local reproductive tract function and causes pain. It is now becoming clear that the endometrium within the uterus in these women differs significantly in function from ‘normal’, and that these anomalies probably precede the development of classical ectopic endometriotic lesions. However, the etiology and mechanisms by which endometriosis arises are still far from certain. This review attempts to address the extensive, but fragmented, evidence that demonstrates widespread molecular disturbances in endometrial function and microstructure underlying this complex condition. The review also addresses some of the novel concepts that are being raised by these exciting new discoveries.

BACKGROUND: Dendritic cells (DCs) are specialized antigen presenting cells that are highly involved in the stimulation and modulation of the immune response within mucosal surfaces, including the female reproductive tract. DCs have been poorly characterized in the non-pregnant endometrium. METHODS: Hysterectomy specimens were obtained from premenopausal women (n = 49) with histoloigically normal endometrium. Endometrial sections were stained immunohistochemically using antibodies for monoclonal mouse anti-human CD1a and CD83, two markers which are specific for populations of immature and mature DCs, respectively. RESULTS: There was a significantly higher density of endometrial CD1a+ DCs than CD83+ DCs throughout the menstrual cycle (P < 0.001). The density of CD1a+ and CD83+ DCs did not vary between the fundus and isthmus of the uterus. There was a significant increase in the density of CD1a + DCs, but not CD83 + DCs, in the basal layer of the endometrium through the phases of the menstrual cycle. The density of CD83+ was significantly greater in the basal layer compared with the functional layer during both the proliferative (P = 0.004) and secretory phases (P = 0.001), whereas for CD1a+ DCs, the greater density in the basal layer was only observed in the secretory phase (P < 0.001). CONCLUSIONS: The highly coordinated cyclical changes in DC populations during the normal menstrual cycle reported in this study may be important for local regulatory mechanisms relevant to menstruation and implantation; alterations in this normal profile may contribute to the development of disturbances of function, fertility and even benign gynaecological disease.

BACKGROUND: The Medicare Modernization Act (MMA) has provided an opportunity for quality improvement organizations (QIOs) to partner with Medicare Part D plan sponsors. These new relationships have developed into a set of diverse projects, each approved by the Centers for Medicare Medicaid Services. OBJECTIVE: To provide information about the scope of the projects being conducted by the QIOs and their partners. SUMMARY: The document describes a variety of quality improvement projects addressing medication use by beneficiaries enrolled in Medicare Part D.Private Medicare QIO contractors are implementing these projects in each state. Descriptions of each project were developed by individual QIOs with the assistance of lead staff for the Physician Practice/Pharmacy QIO SupportCenter for all QIOs nationwide. These projects vary in their complexity, in the quality measures used, and in the clinical processes and economic impact they seek to improve. The summaries in this supplement were prepared 6 months into ...

Spatial processes in pollination biology are poorly understood, especially at levels above that of the local population. For example, little is known about how pollinators and pollen move among populations, although there is evidence that such movement can exceed what is predicted from intrapopulational movement. We explored pollination success in experimental isolates of the bumblebee- and hummingbird-pollinated wildflower Delphinium nuttallianum. We established a total of 15 arrays of potted plants isolated by 50-400 m from ten natural "source" populations, as well as control arrays embedded within each source. Flowers on potted plants were emasculated, so any pollen received could be assumed to come from source populations. A total of 69 h of observation suggested that pollinators were somewhat less abundant in isolates than in controls, but visited more plants and flowers once within an isolate. Consistent with this, 82.1% of all flowers in isolated arrays received pollen, versus 87.7% in controls. Mean receipt was more than 100 pollen grains per flower in most arrays, and seed set in isolates and controls respectively averaged 69.8% and 74.3% of ovules. Furthermore, pollen receipt in isolates declined relatively slowly with distance from the source. We conclude that pollinators of D. nuttallianum often will fly up to 400 m among populations, and that substantial pollination ensues. Thus isolated populations of this species often belong to metapopulations in terms of pollen dispersal, with important consequences for genetic differentiation, and potential implications for the management of endangered plant species.

Endometrial polyps are benign lesions frequently identified in women with infertility or abnormal uterine bleeding in the reproductive and postmenopausal phases We report the striking observation that the numbers of activated mast cells expressing tryptase are increased more than sevenfold throughout the cycle in endometrial polyps (n = 20) compared with normal endometrium. This novel finding has important implications for growth, development, and symptoms associated with polyps in many different tissues.

Research was conducted by staff of the American Pharmaceutical Association (APhA), the national professional society of pharmacists, to inform the APhA policy process. Findings indicate that many consumers view products that were switched from prescription to over-the-counter (OTC) status as being more effective than products that have not previously been designated as prescription drugs, and they prefer to purchase switched products. Consumer purchases of switched products replace utilization of prescription drugs, physician visits, and laboratory services, but no data were found concerning increased utilization that may eventually result from foregone or delayed medical attention. Evidence exists that many comparatively well-informed consumers are knowingly using these products in a manner inconsistent with their labeling, and are using complementary and alternative medications for which labeling is uninformative in lieu of either prescription or OTC products. A need exists for more and better-quality pharmaceutical care services from pharmacists to support informed self-care decision making by consumers.

Objetivo: Valorar si el gradiente alveolo-arterial de oxígeno (Grad[A-a]O2) ayuda a confirmar la influencia de la PEEP en la PaFi (PaO2/FiO2). Diseño: Estudio observacional; usamos una regresión lineal para realizar un estudio multivariable y mejorar la fórmula de la PaFi teniendo en cuenta la PEEP. Ámbito: Hospital terciario. Pacientes: Todos los pacientes de cuidados intensivos, con o sin daño pulmonar. Variables de interés principal: Registramos los antecedentes personales, juicio clínico, datos durante ingreso en UCI, puntuaciones de severidad en el primer día y durante evolución; dos variables calculadas: PaFi and Grad(A-a)O2. Resultados: Un total de 956 patientes incluidos; 63,9% hombres; edad mediana 68 años. Primer día, 31,8% no tienen ventilación mecánica (VM), 13,1% tienen VM no invasiva y 55,1% VM invasiva. PaFi: 32,9% 0-200, 32,2% 201-300, y 34,8% >300. PEEP: 0-5 69,8%, 6-10 27,5% y >10 2,6%. Observamos una correlación (Pearson) entre el Grad(A-a)O2 y la PaFi de -0,84 (p300. PEEP values: 0-5 69.8%, 6-10 27.5% and >10 2.6%. We observed a correlation (Pearson) between Grad(A-a)O2 and PaFi of -0.84 (p

Resumen Objetivo Evaluar la frecuencia de la trombocitopenia grave (TCPG) (≤50.000/μl) en las primeras 24 horas en pacientes con sindrome de disfuncion multiorganica (SDMO) y los factores asociados a su aparicion. Diseno Estudio retrospectivo, con diseno observacional. Ambito Unidad de cuidados intensivos (UCI) medico-quirurgica de un hospital de nivel III. Pacientes Aquellos con disfuncion de al menos dos organos, segun criterios SOFA; se excluyen neurocriticos y politraumatizados. Variables de interes Antecedentes personales, medicacion habitual, situacion funcional basal, datos de filiacion, puntuaciones de gravedad en UCI, datos de la disfuncion multiorganica, evolucion UCI y datos hospitalarios. Resultados Se incluyeron 587 pacientes. El 6,3% (37 pacientes) presentaban TCPG durante el primer dia de ingreso. El 64,6% eran hombres; la mediana de edad fue 69 (56-77) anos; al ingreso, SOFA 8 (5-10); APACHE II 18 (13-24); APACHE IV 59 (46-73); 32,5% son quirurgicos. Durante su evolucion 79,9% necesitaron ventilacion mecanica y el 71,4% requirio farmacos vasoactivos. Estancia en UCI 4 (2-10) dias; estancia hospitalaria 18 (9-35) dias. El 29,2% fallecieron en UCI. El 11,7% desarrollaron durante su ingreso en UCI TCPG. En el analisis multivariable los principales determinantes de la aparicion de la trombocitopenia al ingreso fueron los antecedentes de ingreso hospitalario en el ultimo ano, el peor valor de bilirrubina y albumina sanguinea y la sepsis. Conclusion La prevalencia de TCPG en pacientes criticos con SDMO durante el primer dia de estancia en UCI es del 6,3%. Los factores asociados son: la presencia o no de ingresos hospitalarios en el ultimo ano, los niveles de albumina y bilirrubina y la sepsis.

Amino acids (AAs) are crucial for the developing conceptus prior to implantation. To provide insights into the requirements of the bovine embryo, we determined the AA composition of the uterine fluid. At days 12, 15, and 18 post-estrus, the uteri of synchronized pregnant and non-pregnant Simmental heifers were flushed for the analysis of 41 AAs and their derivatives by liquid chromatography–tandem mass spectrometry. The ipsilateral endometrium was sampled for quantitative PCR. In addition to a pregnancy-dependent increase of the essential AAs (PSLC15A3) were significantly increased at day 18 of pregnancyin vivo. In addition,SLC15A3was predominantly stimulated by trophoblast-derived interferon-τ in stroma cells of anin vitroco-culture model of endometrial cells. Our results show an increased concentration of AAs most likely to optimally provide the elongating pre-attachment conceptus with nutrients.

A well-balanced immunological interaction between mother and the semi-allogenic embryo is of particular importance. The objective of the present study was to analyse mechanisms of immune tolerance in bovine pregnancy during peri-implantation. Simmental heifers inseminated with either cryopreserved spermatozoa or seminal plasma were killed 12, 15 or 18 days after oestrus. Uteri were flushed for the recovery of conceptuses and the ipsilateral intercaruncular endometrium was sampled for gene expression analysis. Indoleamine 2,3-dioxygenase (IDO) mRNA, coding for the initial enzyme of the kynurenine pathway, was 18-fold (P

Bovine trophoblast cells release interferon-τ (IFNT), a type I IFN, as the pregnancy recognition signal. Since type I IFNs exert growth inhibitory and proapoptotic actions, the effect of the conceptus on components of the apoptosis pathways was determined in the bovine endometrium during the periimplantation period. Uteri of Simmental heifers were flushed post mortem at days 12, 15, and 18 of cycle or pregnancy for the recovery of conceptuses and the sampling of ipsilateral endometrial tissue at slaughter for quantitative RT-PCR, immunohistochemistry, caspase activity and TUNEL assays. Endometrium samples of pregnant animals revealed increased transcript levels for the proapoptotic genes XAF1 (day 15: 2.9-fold; day 18: 15.1-fold; p = 0.005) and CASP8 (day 18: 2.4-fold; p = 0.007). The mRNA expression increased significantly with the day of the cycle for the proapoptotic genes FASLG, TNFSF10, TNF and TNFSF1A (p = 0.004, p = 0.006, p = 0.001 and p = 0.007) and the antiapoptotic gene BIRC4 (p = 0.03). We detected high amounts of FASLG transcripts in day 18 conceptuses (16-fold higher than day 18 endometria). This finding was validated at the protein level by immunohistochemistry. To further analyse the endometrial activation of the caspase cascade, the activities of initiator caspase 8 and effector caspases 3/7 were determined luminometrically. No difference between pregnant and cyclic animals was found for either caspase activity. Additionally, a TUNEL assay showed no increase of apoptotic cells in the pregnant endometrium. In conclusion, although the bovine conceptus induces the expression of proapoptotic genes, neither an activation of a caspase cascade nor an increase of apoptotic cells was noticed. These results suggest inhibitory mechanisms preventing endometrial cells from programmed cell death.

Endometriosis is an inflammatory condition, characterized by the presence of endometrial-like tissue outside the uterus. The immune system provides a defence mechanism in response to foreign pathogens, and macrophages play important roles in this response. Activation of macrophages has been reported in peritoneal fluid and ectopic endometriotic lesions; however, controversy exists regarding the composition and function of macrophage populations in eutopic endometrium of women with and without endometriosis. This study aimed to quantify macrophages in eutopic endometrium of women with and without endometriosis, during the early, mid and late proliferative and menstrual phases of the cycle.Paraffin-embedded endometrial curettage blocks were selected from pathology archives. Seventy-six specimens from women with and without endometriosis were analysed using standard immunohistochemical techniques with CD68-PGM1 (phosphoglucomutase 1) clone antibody. Macrophages were counted according to their morphology over several fields of view.A significant increase in macrophage cell numbers was shown in eutopic endometrium in women with endometriosis (mean +/- SD, 182.7 +/- 72.9/mm(2)) during all stages of the proliferative phase compared with normal controls (101.6 +/- 53.4/mm(2); P0.001). Significant increase in macrophage density occurred in the control group during the mid-menstrual phase, Days 3-4 (P0.01), which was not observed in women with endometriosis.This study further supports an association between immune changes in eutopic endometrium and presence of endometriosis. However, it remains uncertain if eutopic immune changes are primary or secondary occurrences.

Prostate-specific membrane antigen (PSMA) is a type 2 integral membrane glycoprotein that serves as an attractive target for cancer immunotherapy by virtue of its abundant and restricted expression on the surface of prostate carcinomas and the neovasculature of most other solid tumors. However, relatively little is known about the molecular structure of this target. Here, we report that PSMA is expressed on tumor cells as a noncovalent homodimer. A truncated PSMA protein, lacking transmembrane and cytoplasmic domains, also formed homodimers, indicating that the extracellular domain is sufficient for dimerization. PSMA dimers but not monomers displayed a native conformation and possessed high-level carboxypeptidase activity. A unique dimer-specific epitope was identified by using one of a panel of novel mAbs. When used to immunize animals, dimer but not monomer elicited antibodies that efficiently recognized PSMA-expressing tumor cells. These findings on PSMA structure and biology may have important implications for active and passive immunotherapy of prostate and other cancers.

It was previously assumed that the import of cytoplasmically synthesized precursor proteins into mitochondria occurs through a single structure spanning both outer and inner membranes at contact sites. Based on recent findings, however, the two membranes appear to contain independent translocation elements that reversibly cooperate during protein import. This feature makes it difficult to generate a means of isolating a fully integrated and functional translocation complex. To study these independent translocases in vitro and in vivo , we have constructed a chimeric protein consisting of an N-terminal authentic mitochondrial precursor (delta 1 -pyrroline-5-carboxylate dehydrogenase) linked, through glutathione S -transferase, to IgG binding domains derived from staphylococcal protein A. This construct becomes trapped en route to the matrix, spanning both outer and inner membranes in such a way that the entire signal-less delta 1 -pyrroline-5-carboxylate dehydrogenase moiety reaches the matrix, while only the folded protein A domain remains outside. During in vivo import of this precursor, outer and inner membranes of yeast mitochondria become progressively “zippered” together, forming long stretches of close contact. Using this novel intermediate, the outer and inner mitochondrial membrane channels, which normally interact only transiently, can be tightly joined (both in vitro and in vivo ), forming a stable association. This suggests a method for isolating the functional translocation complex as a single entity.

Water samples from rivers, streams, ponds, and activated sewage were tested for the presence of bacteria which utilize 2,4-dichlorophenoxyacetic acid (2,4-D) as a sole source of carbon. Seventy percent of the attempted enrichments yielded pure cultures of 2,4-D-metabolizing bacteria. All but 1 of the 30 isolates were gram-negative rods, all but 2 were motile, and all were nonfermentative and oxidase and catalase positive. Nine isolates had DNA guanine-plus-cytosine values of 61.1 to 65 mol%. One isolate had a 67 mol% guanine-plus-cytosine value. The results suggest that these 2,4-D-metabolizing bacteria belong to the genus Alcaligenes. Fourteen of 23 isolates contained one or more detectable plasmids of about 20, 60, or 100 megadaltons. HindIII restriction fragment patterns showed these plasmids to be different from each other with one exception. Very similar restriction fragment patterns were revealed with a plasmid isolated from an Alcaligenes eutrophus strain obtained from Australia (pJMP397) and in an Alcaligenes sp. isolated in Oregon (pEML159). These two plasmids were about 56 megadaltons, had the same guanine-plus-cytosine value, were transmissable, and coded for 2,4-D metabolism and resistance to HgCl2. Hybridization of these two plasmids was demonstrated by using nick-translated 32P-labeled pJMP397. The vector pBR325 was used to clone HindIII fragments from pEML159. One cloned fragment of 14.8 megaldaltons expressed in Escherichia coli the ability to release 14CO2 from 2,4-D labeled in the acetate portion.