Lanthanoid analysis in seawater by seaFAST-SP3™ system in off-line mode and magnetic sector high-resolution inductively coupled plasma source mass spectrometer.

Analysis of lanthanoids in seawater is challenging due to the complex matrix (∼35 g L-1 TDS) and low dissolved concentrations (in ng L-1). A 4-step strict analytical protocol and state-of-the-art technology were implemented and validated in this study. The 4-steps method involves the 1) sample filtration and acidification (pH<2); 2) pre-concentration by the matrix separation system, 3) off-line injection of the eluted sample; and 4) determination of lanthanoids by high-resolution inductively coupled plasma mass spectrometer (HR-ICP-MS). Since there are no certified values for lanthanoids in seawater are available, the method validation was done by analyzing SLEW-3 (estuarine water reference samples) and comparing with other reports and artificial seawater (100 ng L-1 lanthanoid multi-element standard solutions). SLEW-3 recovery varied from 78.6% to 106% and in artificial samples it ranged from 87 to 110%. Low recovery can be explained by complex organic in seawater, because the UV oxidation was not performed in the acidified samples. The variation was ≤10%, except for Gd, Tb, and Yb (11-13.75%). Blanks varied between 0.01 and 0.07 ng L-1, except for La and Ce (0.13-0.21 ng L-1). Blanks represent <5% SLEW-3 values and <1% synthetic seawater. The procedural detection limit varied from 0.01 to 0.03 ng L-1.•Lanthanoids as geochemical tracers in seawaters•A 4-step strict analytical protocol and state-of-the-art technology for lanthanoids analyses in seawaters•Sample pre-concentration system for matrix separation for the detection of ultra-low lanthanoids levels.

Gender differences in ancestral contribution and admixture in Venezuelan populations.

The origin of the contribution of uniparental heritage were analyzed in 615 samples of individuals proceeding from 13 towns classified according to historic differences in their emergence and development as African-derived, European-derived, and admixed/urban. Mitochondrial and Y-chromosome haplogroups were identified by PCR-restriction fragment length polymorphism. The results were compared with previous estimates of admixture made with autosomal markers and with historic aspects. The results show a predominantly indigenous genetic contribution through the female, being more prevalent in urban populations; the African contribution, although dispersed, presents a larger concentration in the African-derived towns, whereas the European contribution is limited to populations with this origin, reflecting isolation and the conservation of the distribution pattern of genes of the Colonial era. With regard to admixture through males, it is almost exclusively of European origin, whereas the African contribution is basically concentrated in the African-derived towns, and the Amerindian lineages are almost nonexistent. The genome of paternal heredity, as opposed to the autosomal and the mitochondrial, shows a homogeneous pattern of admixture that is independent of the origin of the population studied, suggesting that European genes have been introduced into the Venezuelan population through male immigrations, whereas the indigenous contribution has been preserved in the Venezuelan genetic pool through the women. These results provide evidence of the heterogeneity in the genetic origin of the Venezuelan population, which should be taken into account in forensic and epidemiologic genetic studies.

Geogenic lanthanoid signature in coastal and marine waters from the southern Gulf of California.

Lanthanoids in the southern Gulf of California (GC) seawater are reported for the first time. Lanthanoids showed differences between peninsular and continental coastline, coastal or marine ecosystems, and dry or rainy season. The chondrite-normalized values showed high variability but followed a same pattern. Light lanthanoids were more enriched than heavy ones. Values of ∑Ln and La/Lu were higher in continental than peninsular coastlines, coastal than adjacent marine ecosystems, and rainy than dry season. Differences were related to the lithology and perturbation degree of the ecosystem watersheds. The chondrite-normalized patterns are typical of geological origin. Slightly negative Ce anomaly was related to the low levels of oxygen in water for the oxidation of Ce (III) to Ce (IV) and its posterior scavenging. Negative δEu anomaly is explained by an influx of fluvial and eolian materials from the upper continental, while a positive Eu anomaly related to hydrothermal vent inputs was non-evidenced.

3D automatic quantification applied to optically sectioned images to improve microscopy analysis.

New fluorescence microscopy techniques, such as confocal or digital deconvolution microscopy, allow to easily obtain three-dimensional (3D) information from specimens. However, there are few 3D quantification tools that allow extracting information of these volumes. Therefore, the amount of information acquired by these techniques is difficult to manipulate and analyze manually. The present study describes a model-based method, which for the first time shows 3D visualization and quantification of fluorescent apoptotic body signals, from optical serial sections of porcine hepatocyte spheroids correlating them to their morphological structures. The method consists on an algorithm that counts apoptotic bodies in a spheroid structure and extracts information from them, such as their centroids in cartesian and radial coordinates, relative to the spheroid centre, and their integrated intensity. 3D visualization of the extracted information, allowed us to quantify the distribution of apoptotic bodies in three different zones of the spheroid.

Aplicación de PCR a partir de cultivo en la identificación de subespecies del complejo Mycobacterium Tuberculosis / The use of culture-based PCR for the identification of subspecies of the mycobacterium tuberculosis complex

Los métodos diagnósticos clásicos de tuberculosis (TB) se basan en la utilización de baciloscopía y cultivo. La identificación del agente etiológico desde la positivización del cultivo requiere entre 10 y 15 días, mientras que el empleo de la reacción en cadena de la polimerasa (PCR) disminuye el tiempo a 24 h, lo que permite no solo identificar las subespecies del complejo Mycobacterium tuberculosis (CMTB) sino también diferenciarlas de otras especies ambientales clínicamente importantes (MOTT) facilitando el diagnóstico y tratamiento. El objetivo del presente trabajo fue determinar la utilidad de la PCR en la identificación temprana de las micobacterias pertenecientes al CMTB, a partir de cultivos positivos, de pacientes con sospecha de TB, atendidos en un hospital pediátrico de alta complejidad, durante un período de cuatro años. A cada muestra, se le realizó baciloscopía y cultivo en medio líquido. A los cultivos positivos, una inmunocromatografía lateral (TBIDR) y luego PCR. El 4,6% del total de muestras (510/11.162) pertenecientes a 198 pacientes presentó cultivos positivos. Cuatrocientos veintiseis (84%) correspondieron a muestras respiratorias. El rendimiento de la baciloscopía directa fue del 41% (194/470). Cuatrocientos treinta y ocho (86%) resultaron M. tuberculosis, 21 (4%) Mycobacterium bovis, 7 (1%), M. bovis-BCG y 44 (9%) MOTT. La utilización de medios de cultivos líquidos junto con el empleo de PCR favorecen una rápida orientación microbiológica y constituye una estrategia útil para optimizar el manejo clínico de estas infecciones, desde el punto de vista terapéutico y epidemiológico, especialmente en pediatría (AU)

Classical diagnostic methods for tuberculosis (TB) are based on the use of smear microscopy and culture. The identification of the etiological agent from positive culture requires 10 to 15 days, while the use of the polymerase chain reaction (PCR) reduces the time to 24 h, which allows not only to identify the subspecies of the Mycobacterium tuberculosis complex (MTC) but also to differentiate them from clinically important environmental mycobacteria other than tuberculosis (MOTT), facilitating diagnosis and treatment. The aim of this study was to determine the usefulness of PCR in the early identification of mycobacteria belonging to the MTC, from positive cultures of patients with suspected TB seen in a pediatric tertiary hospital over a 4-year period. For each sample, smear microscopy and culture in liquid medium was performed. Positive cultures were subjected to lateral immunochromatography (TBIDR) and then PCR. Of the total number of samples (510/11,162) belonging to 198 patients, 4.6% showed positive cultures; 426 (84%) were respiratory samples. The direct smear microscopy yield was 41% (194/470). Overall, 438 (86%) were found to be M. tuberculosis, 21 (4%) Mycobacterium bovis, 7 (1%), M. bovis-BCG, and 44 (9%) MOTT. The use of liquid culture media together with the use of PCR favors a rapid microbiological orientation and is a useful strategy to optimize the clinical management of these infections, from a therapeutic and epidemiological point of view, especially in children (AU)

Impact of antibiotic treatment on bacterial resistance rates from patients with hospital-acquired infection.

We assessed the impact of antibiotic administration prior to sample collection on the bacterial resistance rates from patients with nosocomial infection. Every individual susceptibility report was assessed in real time at the bedside of the patient by a team composed of infectious diseases and internal medicine specialists as well as clinical microbiologists for clinical significance and appropriateness of the specimen. The report also stated the kind, source and origin of the infection, history of administration of any antibiotic during the last month prior to sample collection. To evaluate the impact of previous antibiotic administration, resistance rates were calculated separately among the group of patients with and without history of antibiotic treatment. A crude univariate analysis was performed to assess the significance of the differences between groups for every species-antibiotic pair. Patients who had received ciprofloxacin showed significantly higher rates of Escherichia coli resistant to ciprofloxacin, broad-spectrum cephalosporins and gentamicin. A higher rate of methicillin-resistant Staphylococcus aureus was observed in patients who were given gentamicin. A stratified analysis showed that the previous antibiotic administration continued to be a risk factor for increased resistance rates regardless of the hospital ward or the source of the infection. This study demonstrates the influence of previous antibiotic administration on bacterial resistance rates although this fact is barely taken into account by the laboratory when constructing the cumulative susceptibility data. Real time clinical validation of the individual susceptibility reports, performed by a multidisciplinary team prior to the data entering, might be a suitable approach to get more reliable susceptibility rates to guide the rational selection of antimicrobial empirical therapy in patients with hospital-acquired infections who have been given antimicrobial treatment prior to specimen collection.

CAS role in the brain apoptosis of Bufo arenarum induced by cypermethrin.

CAS might have a key role in the apoptosis induced by toxins, acting as anti-apoptotic factor, stimulating the cellular proliferation and the cell contact stabilization. To start to elucidate their role in the brain apoptosis of Bufo arenarum induced by cypermethrin (CY), the expression patterns of CAS and several cell adhesion molecules (CAMs) were established. Bufo arenarum tadpoles of the control and acute bioassay survival at different doses (39, 156, 625 and 2,500 microg CY/L) and times (24, 48, 72 and 96 h) of CY treatment were fixed in Carnoy, embedded in paraffin and sectioned. CAS and CAMs expression was determined by immunofluorescence and immunohistochemistry, respectively. When the bioassay starts, CAS increases suggesting a proliferative or regenerative effect, but decreases when the doses and/or the biocide exposure time increases, suggesting compromise of the cellular cycle control and trigger of an apoptotic wave. However, these neurotoxic mechanisms should not involve degradation of N-cadherin and alpha-catenin, in contrast of beta-catenin and axonal N-CAM180, at least in the initial apoptotic phase. Additionally, an adhesion compensatory mechanism by N-CAM180 is observed in the neuron cell body. These results suggest a dual role of CAS in the cellular cycle control during the CY-induced apoptosis: induction of cell proliferation and stabilization of the cell-cell junctions by modulating CAMs expression.

[Compliance with the metabolic goals in diabetes mellitus treatment in the rural area of Ourense, Spain]. / Cumplimiento de los objetivos de control metabólico en diabetes mellitus en el medio rural de ourense.

BACKGROUND: To evaluate the degree to which the metabolic check-up objectives among Type II diabetes patients were met in rural primary care by comparing them to the values recommended by the American Diabetes Association (ADA) and the Diabetes in Primary Care Study Group (GEDAPS). METHODS: Descriptive cross-sectional study. A sample size was calculated based on the glycosolated hemoglobin (HbA1c) test in 119 individuals. A total of 253 patients with Type II diabetes followed up for at least two years at their healthcare facility took part. The patients in question were selected at random from among 17 medical lists at 11 rural healthcare facilities in the province of Ourense (Spain). Demographic variables, cardiovascular risk factors, pharmacological treatment, self-analyses, eye fundus, tactile sensitivity, HbA1c, lipid profile, blood pressure and body mass index data were taken from the patients' clinical records. RESULTS: A total of 44.3% of those in the sample had undergone an HbA1c test within the immediately prior six-month period, showing a value of under seven percent (7%). A total of 21.2% had a blood pressure of under 130/80, and 19.8% a LDL cholesterol level of under 100 mg/dl. A total of 40.7% of those patients with a LDL cholesterol level over 100 were not undergoing any hypolipemiant treatment. A total of 20.4% of the patients showing high blood pressure readings were not undergoing any blood pressure lowering treatment. On the basis of these three factors, a total of 2.5% met the check-up objectives. Thirty-six percent (36%) were antiaggregated with AAS. CONCLUSIONS: A major deficit is revealed both in the frequency of the check-ups conducted by the healthcare professionals as well as the number of interventions performed for achieving the proposed objectives, showing results far below what is recommended in the clinical practice guidelines.

Alternative indicators for monitoring the quality of a continuous intervention program on antibiotic prescribing during changing healthcare conditions.

We recently published on the impact of a four-phase hospital-wide intervention program designed to optimize the quality of antibiotic use, where a multidisciplinary team (MDT) could modify prescription at the last phase. Because health care quality was changing during the last 5 years (late 1999 to early 2004), we developed certain indicators to monitor the quality of our intervention over time. Different periods were defined as baseline (pre-intervention), initial intervention-active control, pre-crisis control, crisis control, post-crisis control and end of crisis control. Major indicators were rates of prescription modification by the MDT; prescription for an uncertain infection and a novel index formula (RIcarb) to estimate the rationale for carbapenem use. We assessed 2115 antimicrobial prescriptions. Modification of prescription rate was 30% at the beginning and decreased thereafter up to stable levels. Rate of prescriptions ordered for cases of both uncertain infection and unknown source of infection decreased significantly after intervention (i.e. from baseline to active control). In contrast, a doubling of culture-directed prescriptions was observed between these periods. RIcarb values lower and higher than 60% (modal, cut-off) were assumed as carbapenem overuse and underuse, respectively. Overuse was observed at the pre-intervention, while pronounced underuse was shown during the crisis (RIcarb, 45% and 87%, respectively). The present study demonstrates that certain indicators, other than the widely adopted impact outcomes, are a suitable tool for monitoring the quality of a continuous, long-term, active intervention on antimicrobial prescribing practice, especially when applied in a changing healthcare setting.

Alterations induced by E-cadherin and beta-catenin antibodies during the development of Bufo arenarum (Anura-Bufonidae).

E(epithelial)-cadherin is a member of a calcium-dependent family of cell surface glycoproteins involved in cell-cell adhesion and morphogenesis. Catenins are a large family of proteins that connect the cadherins to the cytoskeleton. They are important for cadherin function and for transducing signals involved in specification of cell fate during embryogenesis. The best characterized catenins include alpha-, beta-, gamma-, and p120-catenin. Using specific antibodies, we studied the expression and distribution of E-cadherin, and alpha- and beta-catenin in developmental stages of Bufo arenarum toad. The three proteins were found co-localized in stages 19 to 41 of development. Surprisingly, E-cadherin was the only of these three proteins found earlier than stage 19. To test whether E-cadherin and beta-catenin have a functional role in Bufo arenarum embryogenesis, stage 17 whole embryos were incubated with anti-E-cadherin and beta-catenin antibodies. Both anti-E-cadherin and anti-beta-catenin antibodies induced severe morphological alterations. However, while alterations produced by the anti-beta-catenin antibody, showed some variability from the most severe (neural tube and notochord duplication) to a simple delay in development, the alterations with anti-E-cadherin were homogeneous. These observations suggest a critical role for E-cadherin and beta-catenin in the early embryonic development of the Bufo arenarum toad. Our results are consistent with the developmental role of these proteins in other species. One of the most surprising findings was the blockage with the anti-beta-catenin antibodies on later embryo stages, and we hypothesize that the partial axes duplication could be mediated by the notochord induction.

Efficacy of amoxicillin-sulbactam, given twice-a-day, for the treatment of community-acquired pneumonia: a clinical trial based on a pharmacodynamic model.

The present multicenter study reports the results of a clinical trial, designed on the basis of a pharmacodynamic study published previously (Bantar et al., J. Chemother 2000; 12: 223-227) to assess the efficacy of amoxicillin/sulbactam (875 mg/125 mg), given orally twice-a-day for 7 days in the treatment of patients with community-acquired pneumonia (CAP). Eighty-four evaluable subjects older than 19 years with clinical symptoms and features suggestive of CAP, consulting from June 2000 to March 2002 and meeting the PORT risk class I through III, were enrolled in the study. Mean age (y +/- standard deviation) was 46.7 +/- 16.3 and 62% of the patients had some co-morbidity predisposing for CAP. Several individuals (77.4%) fell into a low-risk class (i.e. PORT I or II) and 22.6% of patients belonged to a moderate-risk class at the start of treatment. Six patients (6.45%) had pneumococcal bacteremia. Streptococcus pneumoniae was the organism most frequently isolated (61.9% of all the patients in whom an etiologic diagnosis was made), followed by Haemophilus influenzae. Clinical success was observed in 97.6% of the patients (confidence interval 95%, 94.3%-100%). Almost all the individuals with clinical success became afebrile within the first 3 days of therapy. Ten patients (11.8%) reported mild or moderate adverse events (especially diarrhea) possibly related to the antimicrobial therapy, but this did not lead to withdrawal from the trial. The results of this study suggest that amoxicillin/sulbactam (875 mg/125 mg) is an efficacious and well tolerated option for treating patients with CAP belonging to a low-moderate risk class and support the use of a short, oral (7-day) b.i.d. regimen.

Expression of N-CAM-180 and N-cadherin during development in two South-American anuran species (Bufo arenarum and Hyla nana).

Cadherins and N-CAM are Ca++-dependent and Ca++-independent cell adhesion molecules respectively. These molecules play a key role in morphogenesis and histogenesis. We determined the spatiotemporal pattern of N-cadherin and N-CAM-180 kDa expression by immunohistochemistry during development in two South-American anuran species (Bufo arenarum, toad and Hyla nana, frog). Both N-cadherin and N-CAM were not detectable during early developmental stages. Expression of N-cadherin appeared between the inner and the outer ectoderm layers at stages 19-20. At stages 24-25, N-cadherin was expressed in the neural tube and the heart. In early tadpoles, N-cadherin expression increased along with the central nervous system (CNS) morphogenesis, and reached its maximum level at metamorphic climax stage. N-Cadherin expression was not uniformly distributed. At stage 42, olfactory placodes and retina expressed N-cadherin. Contrary to N-CAM, the strongly myelinated cranial nerves were not labeled. N-Cadherin was present in several mesoderm derivatives such as the notochord, heart and skeletal muscle. The non-neural ectoderm and the endoderm were always negative. Expression of N-CAM appeared first in the neural tube at stages 24-25 and the level of expression became uniform from pre-metamorphic to metamorphic climax tadpoles. At this latter stage, a clear N-CAM immunolabeling appeared in the nerve terminals of pharynx and heart. N-Cadherin and N-CAM were found mainly co-expressed in the CNS from early tadpole to metamorphic climax tadpole. Our results show that the expression of N-CAM and N-cadherin is evolutionary conserved. Their increased expression during late developmental stages suggests that N-CAM and N-cadherin are involved in cell contact stabilization during tissue formation.

Detection of Poinsettia mosaic virus Infecting Poinsettias (Euphorbia pulcherrima) in Venezuela.

Poinsettia mosaic virus (PnMV), a putative member of the tymoviruses, was detected in several cultivars of vegetatively propagated poinsettias grown in commercial nurseries in Estado Miranda, Venezuela. Symptoms associated with the affected plants consisted of severe mottling and distortion of leaves and bracteoles. The suspect virus was mechanically transmitted to Nicotiana benthamiana. Leaf extracts and thin sections of affected leaf tissue were analyzed by transmission electron microscopy. Spherical virus particles (30 nm diameter) were observed in samples from symptomatic poinsettia plants. Ultrastructural analyses of virus-infected cells revealed aggregates of virus particles in the cytoplasm and central vacuole. The virus was purified twice from infected N. benthamiana, resulting in yields as high as 12 mg/100 g. Dissociated coat protein contained a single 24-kDa protein species. The virus was not serologically related to Carnation mottle, Bean rugose mosaic, Cowpea mosaic, Cucumber mosaic, Pea enation mosaic, Prunus necrotic ringspot, Apple mosaic, Tobacco streak, Maize rayado fino, Tomato ringspot, Bean southern mosaic, Sowbane mosaic, Andean potato latent, Belladona mottle, Scrophularia or Turnip yellow mosaic viruses, but did react positively in enzyme-linked immunosorbent assay and western blot analysis with antiserum (ATCC PVAS-476) to PnMV. Based on these results, the virus is considered to be PnMV. To our knowledge, this is the first report of PnMV infecting poinsettias in Venezuela.

Expression of polysialic acid, alpha- and beta-cantenins in adult toad testis in hibernation stage and after gonadotrophin--releasing hormone (GnRH) treatment.

The Polysialic Acid (PSA), glycosydic moiety of the Neural Cell Adhesion Molecule (N-CAM), and alpha- and beta-Catenins, which mediate interaction between Cadherins and cytoskeletal proteins, participate in cell adhesion phenomena in numerous organs and tissues. We have performed an immunohistochemical analysis, in hibernating toad testis and in GnRH-reactivated hibernating animals. In hibernating toads we could demonstrate PSA-immunoreactivity (PSA-IR) within the seminiferous tubules, in clusters of primary spermatocytes, spermatids and spermatozoa, in follicular and Sertoli cells. PSA-IR was seen in peritubular, Leydig and efferent duct cells. In GnRH-treated toads PSA-IR persists in primary spermatocyte groups. alpha-Catenin is localized in the basal laminae of seminiferous tubules and in Leydig cells of hibernating toads. This did not change after hormonal treatment. In hibernating toads, beta-Catenin was detected only in Leydig cells and within seminiferous tubules on basal spermatocystes and limiting spermatozoa clusters. In GnRH-treated toads, the beta-Catenin-IR was less intense in Leydig cells and vanished within seminiferous tubules.

Dominance patterns in macroalgal and phytoplankton biomass under different nutrient loads in subtropical coastal lagoons of the SE Gulf of California.

Nine macroalgal blooms were studied in five coastal lagoons of the SE Gulf of California. The nutrient loads from point and diffuse sources were estimated in the proximity of the macroalgal blooms. Chlorophyll a and macroalgal biomass were measured during the dry, rainy and cold seasons. Shrimp farms were the main point source of nitrogen and phosphorus loads for the lagoons. High biomasses were found during the dry season for phytoplankton at site 6 (791.7±34.6 mg m(-2)) and during the rainy season for macroalgae at site 4 (296.0±82.4 g m(-2)). Depending on the season, the phytoplankton biomass ranged between 40.0 and 791.7 mg m(-2) and the macroalgal biomass between 1 and 296.0 g m(-2). The bulk biomass (phytoplankton+macroalgal) displayed the same tendency as the nutrient loads entering the coastal lagoons. Phytoplankton and macroalgal biomass presented a significant correlation with the atomic N:P ratio.

Isolation and nucleotide sequence analysis of the of Rhinella arenarum ß-catenin: an mRNA and protein expression study during the larval stages of the digestive tract development.

ß-catenin interacts with several proteins mediating key biological processes, such as cadherin-mediated cell-cell adhesion as well as signal transduction. This work was done to establish the molecular basis and regulation of the formation pattern of cadherin/ß-catenin-mediated adherens junctions, using an animal model of unknown gene sequence, the toad Rhinella arenarum. A Rhinella arenarum ß-catenin homolog was isolated from larval tissue, their sequence compared and analyzed with those of eight other vertebrates using bioinformatics tools. The mRNA and protein expression levels of ß-catenin were determined during the development of Rhinella arenarum digestive tract both by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) and immunohistochemistry-morphometry respectively. Using Xenopus laevis frog specific primers, a fragment 539 bp of Rhinella arenarum toad ß-catenin cDNA was obtained and sequenced. The resulting putative sequence of 177 amino acids showed high similarity at the amino acid level (97%) when compared to other six vertebrates (Xenopus laevis, Xenopus tropicalis, Mus musculus, Rattus norvegicus, Bos taurus and Homo sapiens), with sequences and structural domains characteristic of catenins. Subsequently, using primers specifically designed for Rhinella arenarum nucleotide sequence, ß-catenin-mRNA increasing levels were found during the Rhinella arenarum metamorphosis. Finally, increasing ß-catenin protein expression during development has confirmed the specificity the detection of Rhinella arenarum ß-catenin. Summarizing, we have isolated and sequenced a ß-catenin-homologue sequence from the Rhinella arenarum toad, which is highly conserved between species, and following we have detected ß-catenin mRNA and protein levels during their digestive tract development.

T3 regulates E-cadherin, and ß- and α-catenin expression in the stomach during the metamorphosis of the toad Rhinella arenarum.

The metamorphosis of Rhinella arenarum was induced precociously for 5 days, then blocked for 3 months to evaluate the role of thyroid hormones as modulators of morphoregulatory molecules such as E-cadherin, and ß- and α-catenin during epithelium remodeling. We then performed an immunohistochemical and morphometric study of these molecules in the larval stomach. We show that 3,5,3'-triiodothyronine exerts a positive regulatory effect on E-cadherin and ß- and α-catenin expression in stomach epithelium. This suggests continuous synthesis of E-cadherin, and ß- and α-catenin; synthesis essentially is thyroid hormone-independent during premetamorphosis and early prometamorphosis, but it becomes thyroid hormone-dependent during metamorphic climax.

Role of E-cadherin in epithelial architecture maintenance.

Morphogenesis and architecture of a developing epithelium is controlled by both cell shape and contacts, mediated by spatially and temporally regulated cell adhesion molecules. The authors study if E-cadherin functions as a key factor of epithelial adhesion and epidermal architecture in vivo. They apply whole-mount digital deconvolution microscopy to evaluate three-dimensional (3D) E-cadherin expression during skin morphogenesis of Rhinella arenarum and in a cell adhesion alteration model. Results show morphogenetic changes in the 3D E-cadherin spatiotemporal expression pattern correlated with the increase of E-cadherin and in the number of cells with hexagonal geometry. Alterations in junction-protein phosphorylation showed drastic loss of E-cadherin and beta-catenin in cell-cell contacts and the increase of cytoplasm and nuclear beta-catenin in epidermis, suggesting the activation of the beta-catenin signal pathway. Surprisingly, no changes in cell shape and skin architecture were registered, suggesting that epidermal E-cadherin appears to be involved in signaling rather than cell contact maintenance in vivo.