Podocalyxin Expressed in Antigen Presenting Cells Promotes Interaction With T Cells and Alters Centrosome Translocation to the Contact Site.

Podocalyxin (PODXL), a cell surface sialomucin expressed in diverse types of normal and malignant cells, mediates cellular adhesion to extracellular matrix and cell-to-cell interaction. A previous study reported the expression of PODXL protein on monocytes undergoing macrophage differentiation, yet the expression of this molecule in other antigen presenting cells (APCs) and its function in the immune system still remain undetermined. In this study, we report that PODXL is expressed in human monocyte-derived immature dendritic cells at both the mRNA and protein levels. Following dendritric cells maturation using pro-inflammatory stimuli, PODXL expression level decreased substantially. Furthermore, we found that PODXL expression is positively regulated by IL-4 through MEK/ERK and JAK3/STAT6 signaling pathways. Our results revealed a polarized distribution of PODXL during the interaction of APCs with CD4+ T cells, partially colocalizing with F-actin. Notably, PODXL overexpression in APCs promoted their interaction with CD4+ T cells and CD8+ T cells and decreased the expression of MHC-I, MHC-II, and the costimulatory molecule CD86. In addition, PODXL reduced the translocation of CD4+ T-cell centrosome toward the APC-contact site. These findings suggest a regulatory role for PODXL expressed by APCs in immune responses, thus representing a potential target for therapeutic blockade in infection and cancer.

Genomic Multiple Sclerosis Risk Variants Modulate the Expression of the ANKRD55-IL6ST Gene Region in Immature Dendritic Cells.

Intronic single-nucleotide polymorphisms (SNPs) in the ANKRD55 gene are associated with the risk for multiple sclerosis (MS) and rheumatoid arthritis by genome-wide association studies (GWAS). The risk alleles have been linked to higher expression levels of ANKRD55 and the neighboring IL6ST (gp130) gene in CD4+ T lymphocytes of healthy controls. The biological function of ANKRD55, its role in the immune system, and cellular sources of expression other than lymphocytes remain uncharacterized. Here, we show that monocytes gain capacity to express ANKRD55 during differentiation in immature monocyte-derived dendritic cells (moDCs) in the presence of interleukin (IL)-4/granulocyte-macrophage colony-stimulating factor (GM-CSF). ANKRD55 expression levels are further enhanced by retinoic acid agonist AM580 but downregulated following maturation with interferon (IFN)-γ and lipopolysaccharide (LPS). ANKRD55 was detected in the nucleus of moDC in nuclear speckles. We also analyzed the adjacent IL6ST, IL31RA, and SLC38A9 genes. Of note, in healthy controls, MS risk SNP genotype influenced ANKRD55 and IL6ST expression in immature moDC in opposite directions to that in CD4+ T cells. This effect was stronger for a partially correlated SNP, rs13186299, that is located, similar to the main MS risk SNPs, in an ANKRD55 intron. Upon analysis in MS patients, the main GWAS MS risk SNP rs7731626 was associated with ANKRD55 expression levels in CD4+ T cells. MoDC-specific ANKRD55 and IL6ST mRNA levels showed significant differences according to the clinical form of the disease, but, in contrast to healthy controls, were not influenced by genotype. We also measured serum sgp130 levels, which were found to be higher in homozygotes of the protective allele of rs7731626. Our study characterizes ANKRD55 expression in moDC and indicates monocyte-to-dendritic cell (Mo-DC) differentiation as a process potentially influenced by MS risk SNPs.

Emerging Role of Podocalyxin in the Progression of Mature B-Cell Non-Hodgkin Lymphoma.

Mature B-cell non-Hodgkin lymphoma (B-NHL) constitutes a group of heterogeneous malignant lymphoproliferative diseases ranging from indolent to highly aggressive forms. Although the survival after chemo-immunotherapy treatment of mature B-NHL has increased over the last years, many patients relapse or remain refractory due to drug resistance, presenting an unfavorable prognosis. Hence, there is an urgent need to identify new prognostic markers and therapeutic targets. Podocalyxin (PODXL), a sialomucin overexpressed in a variety of tumor cell types and associated with their aggressiveness, has been implicated in multiple aspects of cancer progression, although its participation in hematological malignancies remains unexplored. New evidence points to a role for PODXL in mature B-NHL cell proliferation, survival, migration, drug resistance, and metabolic reprogramming, as well as enhanced levels of PODXL in mature B-NHL. Here, we review the current knowledge on the contribution of PODXL to tumorigenesis, highlighting and discussing its role in mature B-NHL progression.

Poly(3,4-ethylenedioxythiophene):GlycosAminoGlycan Aqueous Dispersions: Toward Electrically Conductive Bioactive Materials for Neural Interfaces.

UNLABELLED: There is an actual need of advanced materials for the emerging field of bioelectronics. One commonly used material is the conducting polymer poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) ( PEDOT: PSS) due to its general use in organic electronics. However, depending on the application in bioelectronics, PEDOT: PSS is not fully biocompatible due to the high acidity of the residual sulfonate protons of PSS. In this paper, the synthesis and biocompatibility properties of new poly(3,4-ethylenedioxythiophene):GlycosAminoGlycan ( PEDOT: GAG) aqueous dispersions and its resulting films are shown. Thus, negatively charged GAGs as an alternative to PSS are presented. Three different commercially available GAGs, hyaluronic acid, heparin, and chondroitin sulfate are used. Indeed, PEDOT: GAGs dispersions are prepared through an oxidative chemical polymerization in water. Biocompatibility assays of the PEDOT: GAGs coatings are performed using SH-SY5Y and CCF-STTG1 cell lines and with ATP and Ca(2+) . Results show full biocompatibility and a pronounced anti-inflammatory effect. This last characteristic becomes crucial if implanted in the body. These materials can be used for in vivo applications, as transistor or electrode for electrical recording and for all the possible situations when there is contact between electronic circuits and living tissues.

Deciphering the Lipid Architecture of the Rat Sciatic Nerve Using Imaging Mass Spectrometry.

Knowledge on the normal structure and molecular composition of the peripheral nerves is essential to understand their pathophysiology and to select the regeneration strategies after injury. However, the precise lipid composition of the normal peripheral nerve is still poorly known. Here, we present the first study of distribution of individual lipids in the mature sciatic nerve of rats by imaging mass spectrometry. Both positive and negative ion modes were used to detect, identify and in situ map 166 molecular species of mainly glycerophospholipids, sphingomyelins, sulfatides, and diacyl and triacylglycerols. In parallel, lipid extracts were analyzed by LC-MS/MS to verify and complement the identification of lipids directly from the whole tissue. Three anatomical regions were clearly identified by its differential lipid composition: the nerve fibers, the connective tissue and the adipose tissue that surrounds the nerve. Unexpectedly, very little variety of phosphatidylcholine (PC) species was found, being by far PC 34:1 the most abundant species. Also, a rich composition on sulfatides was detected in fibers, probably due to the important role they play in the myelin cover around axons, as well as an abundance of storage lipids in the adipose and connective tissues. The database of lipids here presented for each region and for the whole sciatic nerve is a first step toward understanding the variety of the peripheral nerves' lipidome and its changes associated with different diseases and mechanical injuries.

Tratamiento conservador del adenosarcoma mülleriano de cérvix uterino / Conservative treatment of a müllerian adenosarcoma of the uterine cervix

El adenosarcoma mülleriano de cérvix es un tumor muy poco frecuente que aparece generalmente en mujeres jóvenes, como un pólipo cervical sangrante, con tendencia a la recurrencia local debido a su bajo potencial de malignidad. No está claro cuál es su tratamiento óptimo, pero en casos seleccionados se puede realizar tratamiento conservador para preservar la fertilidad de la mujer, destacando la importancia del seguimiento clínico prolongado. Presentamos el caso de un adenosarcoma mülleriano de cérvix en una mujer de 19 años tratada con una conización extensa (AU)

Müllerian adenosarcoma of the cervix is a rare tumor that usually presents in young women as a bleeding cervical polyp. There is a tendency for local recurrence due to their low malignant potential. The optimal therapy for these tumors is uncertain but conservative treatment can be provided in selected patients to preserve female fertility. Long-term follow-up is of paramount importance. We present the case of a 19-year-old girl who was diagnosed with a Müllerian adenosarcoma of the uterine cervix treated with a wide cervical conization (AU)

Metástasis vaginal del carcinoma de células renales / Vaginal metastasis from renal cell carcinoma

El carcinoma de células renales es un tumor de comportamiento impredecible, con alta tendencia a producir metástasis. Puede afectar la vagina, produciendo hemorragia genital. El tratamiento es la biopsia extirpación de la lesión (AU)

Renal cell carcinoma behaves unpredictably, with a strong tendency to produce metastasis, which can affect the vagina, leading to genital bleeding. Treatment is based on wide local excision of the lesion (AU)

Melanoma multifocal del tracto genital femenino / Multifocal melanoma of the female genital tract

El melanoma maligno del tracto genital femenino es una enfermedad multifocal a un trastorno de los melanocitos de las mucosas. Esto justifica su alta tasa de recurrencias a pesar de las resecciones quirúrgicas radicales con márgenes libres. Presentamos el caso de una paciente con melanomas malignos en vulva, vagina y cérvix, acompañados de melanosis del área genital (AU)

Malignant melanoma of the genital tract is a multifocal disease resulting from a disorder of melanocytes within the mucosa, which explains the high recurrence rate after radical excision with free margins. We report the case of a patient with malignant melanoma of the vulva, vagina and cervix, associated with genital melanosis (AU)

Primeros resultados de la disección del ganglio centinela en los carcinomas epidermoides vulvares / First results in the sentinel lymph node procedure in vulvar squamous cell carcinoma

Objetivos: Conocer nuestros primeros resultados en la disección selectiva del ganglio centinela en el cáncer de vulva, en cuanto a tasa de detección del ganglio, casos de falsos negativos y su relación con la recidiva tumoral o no. Material y métodos: Estudio retrospectivo de los primeros 9 casos de ganglio centinela en carcinomas epidermoides vulvares, realizado entre junio de 2004 y diciembre de 2007. Resultados: Se detectó el ganglio centinela en 8 de las 9 pacientes; la tasa de detección fue del 88,8%. No hubo ningún caso de falso negativo, por lo que el valor predictivo negativo de la técnica fue del 100%. Solamente una paciente con ganglio centinela negativo ha fallecido por cáncer de vulva; esta mujer estaba afectada por anemia de Fanconi. Conclusiones: La técnica del ganglio centinela en los carcinomas epidermoides vulvares es factible y puede ser una alternativa a la linfadenectomía inguinofemoral convencional (AU)

Objective: The aim was to analyse our first results in the sentinel lymph node procedure in vulvar cancer, as regards the detection rate, false negative cases and relation with cancer recurrence or not. Material and methods: Retrospective study of first 9 cases of sentinel lymph node detection in vulvar squamous cell carcinoma, between June 2004 and December 2007. Results: We identified the sentinel node in 8 out of 9 patients (88% detection rate). There were no false negative cases, giving a negative predictive value of 100 %. Only one patient with a negative sentinel node died of vulvar cancer. She also had Fanconi anaemia. Conclusions: Sentinel lymph node detection in patients with squamous cell carcinoma of the vulva is feasible, and can be an alternative to conventional inguinofemoral lymphadenectomy (AU)

Downregulation of NR3A-containing NMDARs is required for synapse maturation and memory consolidation.

NR3A is the only NMDA receptor (NMDAR) subunit that downregulates sharply prior to the onset of sensitive periods for plasticity, yet the functional importance of this transient expression remains unknown. To investigate whether removal/replacement of juvenile NR3A-containing NMDARs is involved in experience-driven synapse maturation, we used a reversible transgenic system that prolonged NR3A expression in the forebrain. We found that removal of NR3A is required to develop strong NMDAR currents, full expression of long-term synaptic plasticity, a mature synaptic organization characterized by more synapses and larger postsynaptic densities, and the ability to form long-term memories. Deficits associated with prolonged NR3A were reversible, as late-onset suppression of transgene expression rescued both synaptic and memory impairments. Our results suggest that NR3A behaves as a molecular brake to prevent the premature strengthening and stabilization of excitatory synapses and that NR3A removal might thereby initiate critical stages of synapse maturation during early postnatal neural development.

Cáncer de vulva recidivante en una paciente con anemia de Fanconi / Recurrent vulvar cancer in a patient with Fanconi anemia

La anemia de Fanconi es una enfermedad autosómica recesiva asociada a defectos congénitos, pancitopenia progresiva y predisposición al cáncer. La leucemia mieloblástica aguda es el tipo de cáncer más frecuente, pero los pacientes con anemia de Fanconi también desarrollan diversos tipos de tumores sólidos a unas edades muy tempranas. En las mujeres, los cánceres de vulva y de cérvix son muy frecuentes. Presentamos el caso de una mujer joven con anemia de Fanconi que desarrolló un cáncer de vulva y discutimos el tratamiento cuando la enfermedad se hizo recurrente

Fanconi anemia is an autosomal recessive disorder associated with congenital abnormalities, progressive pancytopenia, and a predisposition to cancer. The most frequent type of cancer is acute myeloid leukemia, but patients with Fanconi anemia also develop various types of solid tumors at remarkably young ages. In women, cervical and vulvar cancer are highly common. We present a case of vulvar cancer in a young woman with Fanconi anemia and discuss the treatment of recurrent disease

In vivo calcium imaging from genetically specified target cells in mouse cerebellum.

Genetically encoded fluorescent calcium indicator proteins provide the potential to monitor activity from genetically specified target cells without a need for single cell resolution. Here we report the use of transgenic mice expressing the fluorescent calcium indicator protein GCaMP2 in cerebellar granule cells to image parallel fiber activity transcranially in vivo. We demonstrated reliable measurements of calcium transients from beams of parallel fibers in response to electrical stimulation in the molecular layer through the intact skull. These parallel fiber calcium transients differed from intrinsic postsynaptic autofluorescence signals in their faster kinetics and resistance to blockers of synaptic transmission. Finally, we used 2P laser-scanning microscopy to demonstrate reliable measurements of calcium transients from beams of parallel fibers at high spatial resolution in living mice. We expect that genetically targeted fluorescent calcium indicator proteins along with optical imaging techniques will be instrumental for the construction of macroscopic and microscopic maps of the function of specific brain circuits.

Optical probing of neuronal circuit dynamics: genetically encoded versus classical fluorescent sensors.

During the past few decades, optical methods for imaging activity in networks composed of thousands of neurons have been developed. These techniques rely mainly on organic-chemistry-based dyes as indicators of Ca(2+) and membrane potential. However, recently a new generation of probes, genetically encoded fluorescent protein sensors, has emerged for use by physiologists studying the operation of neuronal circuits. We critically review the development of these new probes, and analyze objectives and experimental conditions in which classical probes are likely to prevail and where the fluorescent protein sensors will open paths to previously unexplored territories of functional neuroimaging.

Activation of cerebellar parallel fibers monitored in transgenic mice expressing a fluorescent Ca2+ indicator protein.

Genetically encoded fluorescent Ca2+ indicator proteins (FCIPs) are promising tools to study Ca2+ signaling in large assemblies of nerve cells. Currently, there are few examples of stable transgenic mouse lines that functionally express such sensors in well-defined neuronal cell populations. Here we report the generation and characterization of transgenic mice expressing an FCIP under the 5' regulatory sequences of the Kv3.1 potassium channel promoter. In the cerebellar cortex, expression was restricted to granule cells. We first demonstrated reliable measurements of Ca2+ transients from beams of parallel fibers and compared the FCIP signals with intrinsic autofluorescence signals. We demonstrate that, in a transgenic line that exhibits a high expression level of the FCIP, autofluorescence signals are negligible and stimulation-induced fluorescence transients represent FCIP signals. Using frontal cerebellar slices we imaged antidromic activation of granule cells following electrical stimulation of parallel fibers and orthodromic activation of beams of parallel fibers following electrical stimulation of granule cells. We found that paired pulse-induced presynaptic Ca2+ transients of parallel fibers are not affected by blockade of N-methyl-D-aspartate receptors.

Melanosis vaginal asociada a melanoma primario de vulva / Benign vaginal melanosis associated to primary melanoma of the vulva. Case report

Presentamos un caso de melanosis vaginal asociada a melanoma de vulva en una mujer de69 años. Las dos lesiones pigmentarias surgen en la paciente de forma independiente y sin relaciónaparente. La mujer había sido tratada quirúrgicamente por un melanoma maligno primitivode vulva con invasión a nivel IV de Clark y metástasis en una adenopatía inguinal. La lesión demelanosis vaginal se desarrolló tres años después de la vulvectomía radical. Además de la descripcióndel caso se realiza una revisión de la literatura y se comenta sus posibles implicacionesclínico-patológicas

A case of vulvar melanosis associated with primary vulvar melanoma in a 69 yr. old caucasianwoman is reported the two pigmentary lesions became manifest in the patient independentlyand had no apparent relationship. Firstly, the patient was treated by radical vulvectomy for a primitivevulvar melanoma, Clark’s stage IV with metastasis in an inguinal adenopathy. Three yearsafter vulvectomy the patient developed the benign vaginal melanosis. A review of the literatureand a discussion and comments on the clinico-pathological implications between both entitiesare included in this paper

Melanosis vaginal asociada a melanoma primario de vulva / Benign vaginal melanosis associated to primary melanoma of the vulva. Case report

Presentamos un caso de melanosis vaginal asociada a melanoma de vulva en una mujer de 69 años. Las dos lesiones pigmentarias surgen en la paciente de forma independiente y sin relación aparente. La mujer había sido tratada quirúrgicamente por un melanoma maligno primitivo de vulva con invasión a nivel IV de Clark y metástasis en una adenopatía inguinal. La lesión de melanosis vaginal se desarrolló tres años después de la vulvectomía radical. Además de la descripción del caso se realiza una revisión de la literatura y se comenta sus posibles implicaciones clínico-patológicas

A case of vulvar melanosis associated with primary vulvar melanoma in a 69 yr. old Caucasian woman is reported the two pigmentary lesions became manifest in the patient independently and had no apparent relationship. Firstly, the patient was treated by radical vulvectomy for a primitive vulvar melanoma, Clark’s stage IV with metastasis in an inguinal adenopathy. Three years after vulvectomy the patient developed the benign vaginal melanosis. A review of the literature and a discussion and comments on the clinico-pathological implications between both entities are included in this paper

Transgenic mice expressing a pH and Cl- sensing yellow-fluorescent protein under the control of a potassium channel promoter.

During the last few years a variety of genetically encodable optical probes that monitor physiological parameters such as local pH, Ca2+, Cl-, or transmembrane voltage have been developed. These sensors are based on variants of green-fluorescent protein (GFP) and can be synthesized by mammalian cells after transfection with cDNA. To use these sensor proteins in intact brain tissue, specific promoters are needed that drive protein expression at a sufficiently high expression level in distinct neuronal subpopulations. Here we investigated whether the promoter sequence of a particular potassium channel may be useful for this purpose. We produced transgenic mouse lines carrying the gene for enhanced yellow-fluorescent protein (EYFP), a yellow-green pH- and Cl- sensitive variant of GFP, under control of the Kv3.1 K+ channel promoter (pKv3.1). Transgenic mouse lines displayed high levels of EYFP expression, identified by confocal microscopy, in adult cerebellar granule cells, interneurons of the cerebral cortex, and in neurons of hippocampus and thalamus. Furthermore, using living cerebellar slices we demonstrate that expression levels of EYFP are sufficient to report intracellular pH and Cl- concentration using imaging techniques and conditions analogous to those used with conventional ion-sensitive dyes. We conclude that transgenic mice expressing GFP-derived sensors under the control of cell-type specific promoters, provide a unique opportunity for functional characterization of defined subsets of neurons.