Neural cell adhesion molecules in rat endocrine tissues and tumor cells: distribution and molecular analysis

The adhesive properties of neural cell adhesion molecules (NCAMs) can be modified by alternative splicing of the primary transcript or posttranslational modifications. In the present study, we describe distinct forms of alternative splicing and posttranslational modification of the extracellular domain of NCAM of various endocrine tissues and derived tumor cells of the rat. Using an antiserum detecting the immunoglobulin-like domains of NCAM as well as a monoclonal antibody recognizing the NCAM-specific polysialic acid (PSA), we observed a similar staining pattern in adrenals, pituitary, and neoplastic endocrine cells. In endocrine tumor cells [pheochromocytoma (PC12), insulinoma (RINA2), and pituitary tumor cells (GH3)], NCAM immunoreactivity was most intense at contact sites between the cells. The immunocytochemical data were substantiated by results of in situ hybridization histochemistry. Specifically, higher levels of NCAM mRNA were detected in the adrenal cortex than in the medulla. In the pituitary, NCAM mRNA was more abundant in the anterior and intermediate lobes than in the neural lobe. The sequence of NCAM mRNAs in endocrine cells was analyzed by polymerase chain reaction and S1 nuclease protection assays. We found that major exons 4-13 of the NCAM mRNA in endocrine tissues and related tumor cell lines were homologous to those in the brain. However, PC12, RINA2, and GH3 tumor cells; normal rat pituitaries; and adrenals contained different amounts of NCAM mRNA with an alternative extra exon, termed VASE (also called pi in mouse) between constitutive exons 7 and 8. In addition, in pituitaries, we detected an alternative exon in splice site a between the constitutive exons 12 and 13, termed a15, with or without an AAG triplett. These sites are thought to be important for the adhesive properties of NCAM. Therefore, these results suggest that modifications of NCAM may be important for adhesive interactions in normal and neoplastic endocrine cells

Leydig cells express neural cell adhesion molecules in vivo and in vitro

The neural cell adhesion molecule (NCAM) polypeptides are expressed by numerous tissues during embryonic development, where they are involved in cell-cell interactions. In the adult, NCAM expression is confined to a few cell types, including neurons and peptide-hormone-producing cells. Here we demonstrate that the Leydig cells of the adult rat, mouse, and hamster testes express NCAM as well. Western blotting showed that an NCAM of approximately 120 kDa was present in the adult testes of all three species investigated. This form was also found in freshly isolated mouse Leydig cells and in Leydig cells after 2 days in culture. After 4 days in culture, mouse Leydig cells expressed additional NCAM isoforms of approximately 140 and 180 kDa, indicating changes in alternative splicing of NCAM primary transcripts. Also, NCAM mRNA of all isoforms, as detected by S1-nuclease protection assays, increased with time in culture. The expression of the cell adhesion molecule NCAM by adult Leydig cells may explain the aggregation of Leydig cells in clusters in rodent testes, which could be a prerequisite for functional coordination of groups of Leydig cells. Furthermore, the presence of this neural and endocrine marker may indicate a closer relationship between Leydig cells and neural and peptide-hormone-producing cells than is considered to exist at the present time

Caracterização física das chapas de partículas com os rejeitos oriundos da cana-de-açúcar e das folhas de bambu

A problemática do descarte correto dos resíduos sólidos gerados nos diferentes processos industriais faz parte das questões ambientais discutidas na atualidade. Nesse sentido, este trabalho visa estabelecer uma nova alternativa na disposição final de dois rejeitos por meio da produção compósitos para chapas de partículas e, a avaliação das propriedades físicas. O enfoque foi dado à reutilização do bagaço da cana-de-açúcar, originário do processamento da Saccharum officinarum, resíduo oriundo da usina Paraíso Bioenergia, produtora de açúcar e álcool, localizada no município de Torrinha-SP; além das folhas caulinares do bambu da espécie Dendrocalamus giganteus. Para tanto, foram produzidas chapas em seis diferentes tratamentos – 100%, 75%, 50%, 40%, 25% e 0% de bagaço de cana, em relação à massa total dos compósitos, sendo completadas com as folhas de bambu trituradas. Tais chapas foram produzidas e distinguidas por meio de ensaios normalizados da norma NBR 14.810-3 (2002), nas quais foram determinados os coeficientes de teor de umidade (variação de 8,8% a 11,94%) e, absorção (56,3% a 71,3%) e massa específica (0,69 g/cm3 a 0,74 g/cm3).In present days, the disposition of solid residues generated in industrial processes is one of the most important environment questions. In this context, the aim of this work is to establish an alternative to employ these residues, producing particleboards, and evaluation of physical property. The mean focus of the word is reusing: sugar cane bagasse, from processing Saccharum officinarum in Paraíso Bioenergia plant (Torrinha, SP), that produces sugar and ethanol; and caulinar leaves of bamboo Dendrocalamus giganteus. Particleboards were produced in six different insume fractions: 100%, 75%, 50%, 40%, 25% e 0% (sugar cane bagasse) related to total composite mass, completed with bamboo caulinar leaves. The panels were evaluated using normative recommendations from NBR14810-3 (2002) to determine moisture content (ranging from 8.8 % to 11.9%), water absorption (56,3 % to 71,3 %), and density (0,69 g/cm3 to 0,74g/cm3

Ontogeny of synaptophysin and synaptoporin in the central nervous system

The expression of the synaptic vesicle antigens synaptophysin (SY) and synaptoporin (SO) was studied in the rat striatum, which contains a nearly homogeneous population of GABAergic neurons. In situ hybridization revealed high levels of SY transcripts in the striatal anlage from embryonic day (E) 14 until birth. In contrast. SO hybridization signals were low, and no immunoreactive cell bodies were detected at these stages of development. At E 14, SY-immunoreactivity was restricted to perikarya. In later prenatal stages of development SY-immunoreactivity appeared in puncta (identified as terminals containing immunostained synaptic vesicles), fibers, thick fiber bundles and ‘patches’. In postnatal and adult animals, perikarya of striatal neurons exhibited immunoreaction for SO; ultrastructurally SO antigen was found in the Golgi apparatus and in multivesicular bodies. SO-positive boutons were rare in the striatum. In the neuropil, numerous presynaptic terminals positive for SY were observed. Our data indicate that the expression of synaptic vesicle proteins in GABAergic neurons of the striatum is developmentally regulated. Whereas SY is prevalent during embryonic development, SO is the major synaptic vesicle antigen expressed postnatally by striatal neurons which project to the globus pallidus and the substantia nigra. In contrast synapses of striatal afferents (predominantly from cortex, thalamus and substantia nigra) contain SY

Prevalence of Fibromyalgia: A Population‐Based Study in Olmsted County, Minnesota, Utilizing the Rochester Epidemiology Project

Objective To estimate and compare the prevalence of fibromyalgia by 2 different methods in Olmsted County, Minnesota. Methods The first method was a retrospective review of medical records of potential cases of fibromyalgia in Olmsted County using the Rochester Epidemiology Project (from January 1, 2005, to December 31, 2009) to estimate the prevalence of diagnosed fibromyalgia in clinical practice. The second method was a random survey of adults in Olmsted County using the fibromyalgia research survey criteria to estimate the percentage of responders who met the fibromyalgia research survey criteria. Results Of the 3,410 potential patients identified by the first method, 1,115 had a fibromyalgia diagnosis documented in the medical record by a health care provider. The age‐ and sex‐adjusted prevalence of diagnosed fibromyalgia by this method was 1.1%. By the second method, of the 2,994 people who received the survey by mail, 830 (27.6%) responded and 44 (5.3%) met the fibromyalgia research survey criteria. The age‐ and sex‐adjusted prevalence of fibromyalgia in the general population of Olmsted County by this method was estimated at 6.4%. Conclusion To the best of our knowledge, this is the first report of the rate at which fibromyalgia is being diagnosed in a community. This is also the first report of prevalence as assessed by the fibromyalgia research survey criteria. Our results suggest that patients, particularly men, who meet the fibromyalgia research survey criteria are unlikely to have been given a diagnosis of fibromyalgia.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/97488/1/21896_ftp.pd

Effects of Eucalyptus pulp refining on the performance and durability of fibre-cement composites

Although Eucalyptus pulp has been widely used in the paper industry, there is limited information concerning its use as reinforcement in fibre-cement composites. The objective of this study was to evaluate effects of mechanical treatment (refining) of the Eucalyptus pulp on fibre properties as well as performance and microstructure of fibre-cement composites. The composites were evaluated before and after accelerated ageing cycles. The refining increased the capacity of Eucalyptus fibres to capture mineral particles, improving the adherence of the fibres with the matrix. This improved fibre-matrix interface led to better mechanical properties at 28 days of cure but higher mineralisation of fibres and consequently increased brittleness of composites after accelerated ageing (soak and dry) cycles. Unrefined fibres maintained the toughness of composites after ageing cycles. This indicates that refining may weaken the fibres thus affecting the mechanical performance (mainly decreasing modulus of rupture and toughness) of composites after ageing cycles. These results are useful for understanding effects of refined fibre conditions (morphology, mechanical strength and surface properties) on mechanisms of fibre-matrix adherence, fibre mineralisation and degradation of fibre-cement composites.FAPESP (05/59072-4, 07/05299-3)FAPEMIGCAPESCNP

Microbiome profiling by Illumina sequencing of combinatorial sequence-tagged PCR products

We developed a low-cost, high-throughput microbiome profiling method that uses combinatorial sequence tags attached to PCR primers that amplify the rRNA V6 region. Amplified PCR products are sequenced using an Illumina paired-end protocol to generate millions of overlapping reads. Combinatorial sequence tagging can be used to examine hundreds of samples with far fewer primers than is required when sequence tags are incorporated at only a single end. The number of reads generated permitted saturating or near-saturating analysis of samples of the vaginal microbiome. The large number of reads al- lowed an in-depth analysis of errors, and we found that PCR-induced errors composed the vast majority of non-organism derived species variants, an ob- servation that has significant implications for sequence clustering of similar high-throughput data. We show that the short reads are sufficient to assign organisms to the genus or species level in most cases. We suggest that this method will be useful for the deep sequencing of any short nucleotide region that is taxonomically informative; these include the V3, V5 regions of the bac- terial 16S rRNA genes and the eukaryotic V9 region that is gaining popularity for sampling protist diversity.Comment: 28 pages, 13 figure

Oregon’s Early Hearing Detection and Intervention Program (EHDI): The First Fifteen Years (1989-2004)

Following unsuccessful efforts in the early 1990’s, using a “high-risk” registry to identify infants with hearing loss, and following the development of practical automated electrophysiologic technology to identify hearing loss in infants, community advocates and the Office of Family Health (OFH) lobbied to introduce universal newborn hearing screening in Oregon. In 1999, the Legislature adopted a compromise bill that mandated screening in all hospitals with more than 200 births annually. When the new law took effect, in July 2000, the proportion of infants screened rapidly rose from 25% to over 94%. In 2003, the Legislature added mandatory individual-level reporting and the OFH established a registry. With these essential elements now in place, a critical task is to ensure complete follow-up of infants failing newborn hearing screening with diagnostic testing and early intervention, if appropriate. Health promoters and educators will play a key role encouraging participation in the Early Hearing Detection and Intervention Program by health professionals and parents

Differential expression of synaptophysin and synaptoporin during pre- and postnatal development of the hippocampal network

The closely related synaptic vesicle membrane proteins synaptophysin and synaptoporin are abundant in the hippocampal formation of the adult rat. But the prenatal hippocampal formation contains only synaptophysin, which is first detected at embryonic day 17 (E17) in perikarya and axons of the pyramidal neurons. At E21 synaptophysin immunoreactivity extends into the apical dendrites of these cells and in newly formed terminals contacting these dendrites. The transient presence of synaptophysin in axons and dendrites suggests a functional involvement of synaptophysin in fibre outgrowth of developing pyramidal neurons. Synaptoporin expression parallels the formation of dentate granule cell synaptic contacts with pyramidal neurons: the amount of hippocampal synaptoporin, determined in immunoblots and by synaptoporin immunostaining of developing mossy fibre terminals, increases during the first postnatal week. Moreover, in the adult, synaptoporin is found exclusively in the mossy fibre terminals present in the hilar region of the dentate gyrus and the regio inferior of the cornu ammonis. In contrast, synaptophysin is present in all synaptic fields of the hippocampal formation, including the mossy fibre terminals, where it colocalizes with synaptoporin in the same boutons. Our data indicate that granule neuron terminals differ from all other terminals of the hippocampal formation by the presence of both synaptoporin and synaptophysin. This difference, observed in the earliest synaptic contacts in the postnatal hippocampus and persisting into adult life, suggests distinct functions of synaptoporin in these nerve terminals