bluuebunny/crossref_metadata_2025_split huggingface dataset:
Correlation of the BRAF V600E mutation in hairy cell leukaemia with morphology, cytochemistry and immunophenotype
Authors: LANGABEER S. E. , O’BRIEN D. , LIPTROT S. , FLYNN C. M. , HAYDEN P. J. , CONNEALLY E. , BROWNE P. V. , VANDENBERGHE E. 
Published: February 2012

SummaryHairy cell leukaemia (HCL) has distinct clinical, morphological and immunophenotypic features with no recurrent cytogenetic or molecular abnormalities reported until the recent description of the BRAF V600E mutation in patients with classical HCL. The incidence of this mutation was sought in 27 patients with either classical HCL or HCL variant by an allele‐specific PCR approach and findings related to morphology, cytochemistry and immunophenotype. A high degree of correlation was noted between the presence of BRAF V600E and established diagnostic criteria in 26/27 patients with HCL/HCL variant. Detection of the BRAF V600E mutation is therefore a useful adjunct in the differential diagnosis of HCL and HCL variant and highlights the value of a multifaceted approach to the diagnosis of this malignancy.
Catabolism and biotechnological applications of cholesterol degrading bacteria
Authors: García J. L. , Uhía I. , Galán B. 
Published: February 2012

SummaryCholesterol is a steroid commonly found in nature with a great relevance in biology, medicine and chemistry, playing an essential role as a structural component of animal cell membranes. The ubiquity of cholesterol in the environment has made it a reference biomarker for environmental pollution analysis and a common carbon source for different microorganisms, some of them being important pathogens such as Mycobacterium tuberculosis. This work revises the accumulated biochemical and genetic knowledge on the bacterial pathways that degrade or transform this molecule, given that the characterization of cholesterol metabolism would contribute not only to understand its role in tuberculosis but also to develop new biotechnological processes that use this and other related molecules as starting or target materials.
Examiner reliability of fluorosis scoring: a comparison of photographic and clinical examination findings
Authors: Cruz‐Orcutt Noemi , Warren John J. , Broffitt Barbara , Levy Steven M. , Weber‐Gasparoni Karin 
Published: February 2012

AbstractObjective: To assess and compare examiner reliability of clinical and photographic fluorosis examinations using the Fluorosis Risk Index (FRI) among children in the Iowa Fluoride Study (IFS).Methods: The IFS examined 538 children for fluorosis and dental caries at age 13 and obtained intraoral photographs from nearly all of them. To assess examiner reliability, duplicate clinical examinations were conducted for 40 of the subjects. In addition, 200 of the photographs were scored independently for fluorosis by two examiners in a standardized manner. Fluorosis data were compared between examiners for the clinical exams and separately for the photographic exams, and a comparison was made between clinical and photographic exams. For all three comparisons, examiner reliability was assessed using kappa statistics at the tooth level.Results: Interexaminer reliability for the duplicate clinical exams on the sample of 40 subjects as measured by kappa was 0.59, while the repeat exams of the 200 photographs yielded a kappa of 0.64. For the comparison of photographic and clinical exams, interexaminer reliability, as measured by weighted kappa, was 0.46. FRI scores obtained using the photographs were higher on average than those obtained from the clinical exams. Fluorosis prevalence was higher for photographs (33 percent) than found for clinical exam (18 percent).Conclusion: Results suggest that interexaminer reliability is greater and fluorosis scores are higher when using photographic compared with clinical examinations.
A multicentre prospective study of post‐traumatic endophthalmitis
Authors: Cornut Pierre‐Loïc , Youssef El Bichara , Bron Alain , Thuret Gilles , Gain Philippe , Burillon Carole , Romanet Jean‐Paul , Vandenesch François , Maurin Max , Creuzot‐Garcher Catherine , Chiquet Christophe ,   for the French Institutional Endophthalmitis Study (FRIENDS) Group
Published: February 2012

Abstract.Purpose:  Study the clinical and microbiological characteristics and the prognostic factors of post‐traumatic endophthalmitis.Methods:  Seventeen eyes were included between 2004 and 2010, with clinical and microbiological data collected prospectively. Conventional cultures and panbacterial PCR were performed on aqueous and vitreous samples.Results:  Clinical signs of endophthalmitis were observed soon after trauma (1.5 ± 2.5 days). Laceration with an intraocular foreign body (IOFB) was noted in 53% of the patients. At admission, all patients had aqueous humour (71%) and/or vitreous (53%) samples. Fifteen patients (88%) underwent a pars plana vitrectomy. Bacteria were identified in 77% of the cases: Staphylococcus epidermidis (n = 5), Streptococcus (n = 4), Bacillus (n = 2), Pseudomonas stuzeri (n = 1), and Streptococcus salivarius and Gemella haemolysans (multibacterial infection, n = 1). Progression toward phthisis was observed in 35% of the cases; 41% of the patients recuperated visual acuity (VA) ≥20/40. A good final visual prognosis (≥20/40) was significantly associated with initial VA better than light perception (0% versus 70%, p = 0.01) and absence of pupillary fibrin membrane (80% versus 20%, p = 0.05). There was no correlation between visual prognosis and age, the type of laceration (corneal or scleral) or presence of an IOFB. We found a statistical trend toward an association between bacterial virulence and poor final VA.Conclusion:  This series showed that better final VA outcomes were associated with initial VA better than light perception, S. epidermidis or culture‐negative cases and absence of retinal detachment during the clinical course.
Regulation of the expression of interleukin‐8 induced by 25‐hydroxycholesterol in retinal pigment epithelium cells
Authors: Catarino Steve , Bento Carla F. , Brito Ana , Murteira Eliana , Fernandes Alexandre F. , Pereira Paulo 
Published: February 2012

Abstract.Purpose:  This study aimed at elucidating the molecular mechanisms involved in the regulation of IL‐8 production by several oxysterols in retinal pigment epithelium (RPE) cells.Methods:  A human cell line from RPE (ARPE‐19) was used to test the role of cholesterol and several oxysterols (25‐OH, 7‐KC and 7β‐OH) in the expression and secretion of IL‐8. Expression of IL‐8 was assessed by real‐time PCR, while IL‐8 secretion was evaluated by ELISA. PI3K‐, MEK1/2‐, ERK1/2‐ and NF‐κB‐specific inhibitors were used to assess the specific role of the several players on the regulation of IL‐8 production by oxysterols. A gene‐reporter assay for AP‐1 activity was also conducted to evaluate the putative role of this transcription factor on IL‐8 expression induced by oxysterols.Results:  Here, we demonstrate that 25‐OH specifically increases transcription and secretion of the cytokine IL‐8 in ARPE‐19 cells. Indeed, treatment of ARPE‐19 with 25‐OH, but not with 7‐KC, 7β‐OH or cholesterol, induced the secretion of IL‐8 from cells. 25‐OH also induced the activation/phosphorylation of ERK1/2 through a mechanism dependent on MEK, ERK1/2 and PI3K kinase activity. Real‐time PCR and ELISA experiments demonstrated that 25‐OH increased transcription and secretion of IL‐8 through a mechanism that is dependent on ERK1/2 and PI3K activity. Furthermore, 25‐OH triggered the activation/phosphorylation of the AP‐1 component c‐Jun and, consistently, increased the transcriptional activity of AP‐1. Additionally, we also found that 25‐OH decreases the levels of IκB and increases the nuclear levels of NF‐κB p65 subunit and that inhibition of NF‐κB activity partially prevents the increased secretion of IL‐8 induced by 25‐OH.Conclusions:  The results presented in this study suggest a role for 25‐OH in inducing IL‐8 production through pathways that are likely to involve AP‐1 and NF‐κB in ARPE‐19 cells. Our data may also provide new molecular targets for the treatment of AMD.
Microcalorimetric studies of hybridoma cells
Authors: Nässberger Lennart , Truedsson Lennart , Monti Mario 
Published: February 2012

In the present study, overall metabolism has been estimated in hybridoma cells by microcalorimetric measurement. Heat production rate was found to be 30–50 pW/cell at cell concentrations 0.65t‐4.5×105/ml. High cell concentrations (1×106 cells/ml) caused unstable power‐curves with an initial high peak and a rapid declining phase, whereas low cell concentrations (0.6−4.5×105 cells/ml) produced steady‐state power‐curves. Oxygen consumption was found to range between 1.5−6.1×10−5 mol 02/cell/min, corresponding to about 80% of the total metabolic activity.The metabolic inhibitors sodium fluoride (50 mM), sodium azide (160 mM) androtenone (0.1 mM) caused a reduction in overall cell metabolism of 60, 55 and 40% respectively.
Immunocytochemical detection of ribulose bisphosphate carboxylase in capsicum plastids
Authors: Cheniclet Catherine , Suire Claude , Carde Jean‐Pierre 
Published: February 2012

SUMMARYRibulose bisphosphate carboxylase (RUBPCase) was localized by fluorescence and gold immunocytochemistry in Capsicum fruits. Chloroplasts of the green fruit are heavily labelled. A positive staining is also obtained with chromoplasts of the ripe rad fruit, but gold labelling is fainter. The presence of reactive RuBPCase in chromoplasts is discussed in relation with the absence of ribosomes in these plastids.
Characterization of a ferritin isolated from the midgut epithelial cells of a homopteran insect, Philaenus spumarius L.
Authors: Collin Olivier , Thomas Daniel , Flifla Mohammed , Quintana Carmen , Gouranton Jean 
Published: February 2012

Crystalline accumulations of ferritin‐like particles are present within the cytoplasm and the nucleus in midgut epithelial cells of the homopteran Philaenus spumarius.A structural study at the electron microscope level reveals that these particles have the morphological characteristics of the ferritin molecule: crystals have a face‐centered cubic structure with a lattice parameter of 14 ± 1 nm; negatively stained isolated particles have the appearance of ferritin; on rotary‐shadowed particles 3 axes of symmetry are clearly seen; image processing performed on selected molecules demonstrates a 4‐fold symmetry.A semiquantitative electron microprobe analysis effected on aggregates of microcrystals in thin sections reveals a high atomic ratio Fe/P.Analyzed by SDS‐PAGE, the protein subunit has a molecular weight of 18,600. The amino acid composition of the protein bears the general characteristics of the ferritin molecule in terms of polar and nonpolar residues. But in terms of sequences, this protein displays a strong dissimilarity to rat liver ferritin as demonstrated with a common amino acid index test and with immunoelectrophoresis experiments.
Analysis by polarizing microscopy of chromosomal structure among dinoflagellates and its phylogenetic involvement
Authors: Cachon Jean , Sato Hidemi , Cachon Monique , Sato Yukiko 
Published: February 2012

Dinoflagellate chromosomes are highly interesting because of their condensed state, their lack of histone, and their ultrastructure reminiscent of that of bacterial nucleoid.With the use of a highly sensitive resolution polarization microscope coupled with electron microscope observations, and by studying and comparing a great variety of species, we confirm the intrinsic nature of birefringence and bring new data about the evolution of interphasic chromosomes among dinoflagellates. Three types are observed: the A‐type, characterized by positive birefringence, condensed state, and lack of basic proteins; the B‐type, with a half positive‐half negative birefringence corresponding to a neck lace chromosome shape and possessing no or very few basic proteins; and the C‐type, which has a negative birefringence because of its loosely gathered filament bundle possessing few basic proteins.
Nucleolar component behaviour in plant cells under different physiological conditions. A morphological, cytochemical and quantitative study
Authors: Moreno Francisco Javier , Rodrigo Rosa M. , Gracia‐Navarro Francisco , Garcia‐Herdugo Gregorio 
Published: February 2012

Different nucleolar components of Allium cepa L. root meristem cells under physiological conditions of proliferation and quiescence were quantitatively analyzed after the application of morphometric and cytochemical techniques.Our data show that quiescent cells have a diminished nucleolar size when compared with proliferating cells. However, the silver stained area in relation to the nucleolar area (fibrillar component) is maintained constant in both physiological conditions. We also detected a smaller number of vacuoles per nucleolar volume unit in the case of low transcriptional activity. Cytochemical results obtained in both physiological conditions allowed us to postulate about the distribution of the acidic nucleolar proteins responsible for the NOR‐silver staining.
Renewal of goblet cell mucus granules during the cell migration along the crypt‐villus axis in rabbit jejunum: an immunolabeling study
Authors: Chambraud Lydie , Bernadac Alain , Gorvel Jean‐Pierre , Maroux Suzanne 
Published: February 2012

A monoclonal antibody against intestinal mucins (5H7) was obtained and used to immunolabel thin frozen sections and Epon‐embedded sections of rabbit jejunum. It recognized a mucin oligosaccharide, the synthesis of which increased during goblet cell migration along the crypt‐villus axis. During the earliest steps in their differentiation, the goblet cells located at the bottom of crypts synthesized mucins devoid of the 5H7 epitope, thus generating unlabeled granules. These unlabeled granules were gradually replaced by more and more labeled granules during the cell maturation. During goblet cell migration along the middle half of the villi, the mucus granules were found to be totally renewed twice. However, some newly formed labeled granules were observed to reach the apical pole of the cells before older unlabeled ones and might have had a faster turnover.At least one glycoconjugate of the goblet cell microvillar membrane also bore the 5H7 epitope. It was rapidly carried from the Golgi apparatus to the apical plasma membrane domain by a transport process that was independent of baseline mucin secretion.
Epithelial cell migration on small intestinal villi in the neonatal rat. Comparison between [3H] thymidine and cytoplasmic labelling after Pu‐citrate ingestion
Authors: Moutairou Kabirou , Fritsch Paul , Meslin Jean‐Claude , Poncy Jean‐Luc , Metivier Henri , Masse Roland 
Published: February 2012

This study compares, in 2‐d‐old rats, the migration rates of epithelial cells on villi of the small intestine, using two labelling methods: a single [3H] thymidine injection; and cytoplasmic labelling by a single ingestion of Pu‐citrate. Histoautoradiography shoed negligible diffusion of Pu after the initial retention, which was mostly confined to the epithelial cells of the villi. However, after sloughing of labelled cells in the intestinal lumen, Pu was reabsorbed by the distal epithelial cells. In segments in which Pu reabsorption was negligible, the migration rates of Pu‐ and 3H‐labelled cells were very close. These rates, expressed in micrometers, were almost constant along the length of the villus, and the Pu and 3H labelling edges reached the top of the villi in about 5 and 7 d, respectively.Once Pu retention had reached its maximum in 9 equal segments cut along the small intestine, tissue counting showed an exponential Pu release of 30–40%d from each segments until the end of the experiment at d16. This constant release might reflect a constant cell migration rate during the period from Pu ingestion until d16.
Size‐related differences in the branching pattern of the motor nerve terminals in triangularis sterni muscle of the mouse
Authors: Tomasi Josep , Fenol Rosa , Santafe Manel , Mayayo Emili 
Published: February 2012

A light microscopy morphometric study was performed in singly innervated synaptic areas of the triangularis sternal muscle of the normal adult Swiss mouse. Investigating mechanisms of the motor nerve growth control, we tested the hypothesis that significant differences in the nerve terminal branching pattern can be detected between different populations of nerve endings classified according to their arborization complexity or size.The main observations of this morphometric study are first, that the mean segment length of the terminal arborization between branch points behaves as an independent variable from the remaining parameters; the mean value of this parameter did not change in nerve endings of differing size and complexity. Secondly, the increase in size of the nerve endings is accompanied by a significant reduction in the mean length of the distal free‐end segments. Results are discussed in the context of the possible regulatory mechanisms governing nerve terminal growth and remodelling.
Localization of barriers to water flow in toad urinary bladder
Authors: Levine Sherman D. , Jacoby Monica 
Published: February 2012

Although it is well accepted that vasopressin (ADH) increases the permeability to water of the toad bladder granular cell's luminal membrane, recent studies have suggested that regulation also takes place at an additional “postluminal” site within the epithelial granular cell. These studies are based upon the observation that a number of experimental maneuvers can alter tissue permeability to water, but do not change the number of particle aggregates observed on the protoplasmic face of the granular cell's luminal membrane with freeze‐fracture electron microscopy. These aggregates are believed by many investigators to mediate the transport of water across the luminal membrane.The dissociation between permeability and aggregate frequency described above has been variously interpreted as the consequence of changes in the permeability of the aggregates themselves, or of changes in the permeability of a “postluminal” barrier that is functionally in series with the luminal membrane.We attempted to distinguish between these 2 possibilities by studying paired toad bladders during 3 protocols that alter vasopressin‐stimulated water flow across the intact tissue without altering aggregate frequency. Estimates of the permeability of postluminal barriers were obtained by exposing the luminal surface to amphotericin B, an antibiotic that forms water‐permeant channels in the luminal membrane.Of the 3 protocols, only diminishing bladder filling volume decreased the water flow elicited by luminal amphotericin B, suggesting that only that protocol indeed decreased the permeability of some postluminal barrier. The other 2 protocols, increasing PCO2 and repeatedly stimulating the bladder with vasopressin, did not alter amphotericin B‐elicited flow, suggesting that postluminal barriers were not altered by these 2 protocols. We thus conclude that a decrease in luminal membrane aggregate permeability or a decreased frequency of fusion of aggregate‐rich tubules to the luminal membrane mediates the decreased flow‐response in the latter 2 protocols, and not modulation of a postluminal membrane barrier.
Isolation of frog urinary bladder plasma membranes with polycation coated beads
Authors: Valenti G. , Casavola V. , Svelto M. 
Published: February 2012

It is now generally accepted that the increase in water permeability induced by antidiuretic hormone (ADH) in responsive epithelia is accompanied by the insertion of specific structures in the apical membrane of epithelial cells. There are strong indications that these particles, probably proteic in nature, represent water channels.In order to evaluate the nature and role of such proteins, plasma membranes were isolated by the affinity chromatography technique. The method is based on the firm attachment of the external face of the membrane to polycations covalently bound to the surface of polyacrylamide beads, followed by shearing of the rest of the cells. Maximal binding of epithelial cells to beads was achieved in a medium of low ionic strength and pH 5.2 (i.e. sucrose‐MES buffer). By this procedure plasma membranes were obtained from both cAMP‐stimulated cells and control cells.Membranes isolated on beads were enriched in the activity of typical membrane marker enzymes (LAP; H+ ATPase; Na+, K+ ATPase) with respect to a whole cell homogenate, whereas contamination of plasma membrane fraction by endoplasmic reticulum, lysosomes, and mitochondria was relatively low.Analysis by SDS polyacrylamide gel electrophoresis showed an interesting difference between cAMP‐treated and control samples.
Cellular permeation pathways in a leaky epithelium: the human amniochorion
Authors: Hardy Marcos A. , Leonardi Ronald T. , Scheide John I. 
Published: February 2012

The presence of cellular permeation pathways in human fetal membranes at term was evaluated. Electrical parameters (transepithelial potential [TEP], and conductance [Gt], and intracellular potentials [cell PD]), and water and urea diffusional coefficients (Pdw, Pdu), were determined in Ussing‐like chambers.In amniochorion, the TEP was practically 0 (0.1±0.03 mV), and the Gt very high (144±14 mS/cm2). The Cell PD of amnion cells was −37±3 mV. Increasing the [K+] of the amniotic perfusate between 5.8 and 125.8 mM depolarized the cells with a slope of 23 mV. The deletion of Na+ hyperpolarized the cells, whereas amiloride and ouabain depolarized them.The Pdw and Pdu were determined in intact amnion and chorion and in their epithelial cell layers. The Pdw/Pdu ratio in amnion was 4.0, and 7.0 in its cell layer; the ratio in chorion was 2.5, and 3.3 in its cell layer.The amniochorion is a leaky structure, but its cellular layers possess definite transcellular permeation pathways. The ionic conductances in amnion cells are complex, with the Cell PD being determined by at least K+ and Na+ conductances, and ouabain‐ and amiloride‐sensitive pathways. The amnion is a more effective diffusional barrier to water and urea than chorion is; its diffusional characteristics are comparable to those of nystatin‐treated lecithin: cholesterol bilayers and the membranes of human erythrocytes.
Coupling of volume and Na+ transport in frog skin epithelium
Authors: Tang Cherng‐Shing R. , Peterson‐Yantorno Kim , Civan Mortimer M. 
Published: February 2012

Whole skins and isolated epithelia were bathed with isotonic media (‐ 244 mOsm) containing sucrose or glucose. The serosal osmolality was intermittently reduced (to ‐ 137 mOsm) by removing the nonelectrolyte. Transepithelial and intracellular electrophysiological parameters were monitored while serosal osmolality was changed. Serosal hypotonicity increased the short‐circuit current (Isc) and the basolateral conductance, hyperpolarized the apical membrane (Φmc), and increased the intracellular Na+ concentration. The increases in apical conductance and apical Na+ permeability (measured from Goldman fits of the relationship between amiloride‐sensitive current and Φmc) were not statistically significant.To verify that the osmotically induced changes in Isc were mediated primarily at the basolateral membrane, the basolateral membrane potential of the experimental area was clamped close to 0 mV by replacing the serosal Na+ with K+ in Cl−‐free media. The adjoining control area was exposed to serosal Na+. Serosal hypotonicity produced a sustained stimulation of Isc across the control, but not across the adjoining depolarized tissue area.The current results support the concept that hypotonic cell swelling increases Na+ transport across frog skin epithelium by increasing the basolateral K+ permeability, hyperpolarizing the apical membrane, and increasing the electrical driving force for apical Na+ entry.