bluuebunny/crossref_metadata_2025 huggingface dataset:
Conservative surgical debridement as a burn treatment: Supporting evidence from a porcine burn model
Authors: Wang Xue‐Qing , Kempf Margit , Liu Pei‐Yun , Cuttle Leila , Chang Hong‐En , Kravchuk Olena , Mill Julie , Phillips Gael E , Kimble Roy M.
Published: August 2010

In thermal deep‐dermal burns, surgical debridement is normally used in conjunction with skin grafting or skin substitutes and debridement alone as a burn treatment is not usually practiced. The current study addresses whether or not debridement alone would enhance burn wound healing on small deep‐dermal‐partial thickness burns. This was a prospective and blinded experimental trial using a porcine deep‐dermal‐partial thickness burn model. Four burns, approximately 50 cm2 in size, were created on each of eight pigs. Two burns from each pig were immediately surgically debrided and the other two were not debrided as the internal control. Hydrate gel together with paraffin gauze were used to cover the burns for four pigs and silver dressings for the other four. Clinical assessment of wound healing was conducted over a 6‐week period. Skin samples were collected at the end of the experiment and histopathological evaluation was performed. The results show thinner scar formation and lower scar height in the debrided compared with nondebrided wounds in the hydrate gel/paraffin gauze groups. There were no statistically significant differences in wound healing assessment between the debrided and nondebrided wounds dressed with silver dressings. This study provides supporting evidence that immediate debridement with an appropriate dressing and without skin grafting may promote wound healing, suggesting its potential benefit for clinical patients.
A poly‐herbal formulation accelerates normal and impaired diabetic wound healing
Authors: Gupta Asheesh , Upadhyay Nitin K. , Sawhney R.C. , Kumar Ratan
Published: August 2010

In the present study, a poly‐herbal formulation (PHF) was prepared by combining the aqueous lyophilized leaf extracts of Hippophae rhamnoides L. and Aloe vera L. and the ethanol rhizome extract of Curcuma longa L., in an optimized ratio (1 : 7 : 1). The efficacy of PHF treatment was studied in normal and impaired diabetic rats using a full‐thickness cutaneous wound model. Topical PHF treatment increased cellular proliferation and collagen synthesis at the wound site in normal rats, as evidenced by the significant increase in DNA, total protein, hydroxyproline, and hexosamine contents in comparison with a positive control treated with a povidone–iodine ointment. The histological examinations and matrix metalloproteinases expression also correlated well with the biochemical findings, confirming the efficacy of PHF in normal wounds. In the streptozotocin‐induced diabetic rats, PHF treatment increased hydroxyproline and hexosamine content. A faster wound contraction was also observed in PHF‐treated normal and diabetic rats. The PHF also promoted angiogenesis as evidenced by an in vitro chick chorioallantoic membrane model and in vivo up‐regulated vascular endothelial growth factor expression. The results suggest that PHF possesses significant wound healing potential in both normal as well as chronic diabetic wounds.
S163 is critical for FXYD5 modulation of wound healing in airway epithelial cells
Authors: Miller Timothy J. , Davis Pamela B.
Published: August 2010

The FXYD family, which contains seven members, are tissue specific regulators of the Na,K‐ATPase. Increased expression of FXYD5, a cancer–cell‐associated membrane glycoprotein, has been associated with increased cell motility and metastatic potential. To better understand how FXYD5 may modulate cell motility, we analyzed S163, a conserved residue in all FXYD family members located in the C‐terminus. Ectopic expression of human FXYD5 S163 mutants in HEK 293 cells showed that negative charge at S163 (S163D) decreased membrane localization, assessed by immunofluorescence. Coimmunoprecipitation studies revealed decreased FXYD5/Na,K‐ATPase interaction for S163D compared with wild‐type or S163A mutants. Interestingly, FXYD5 overexpression induced expression of vimentin, a marker of epithelial–mesenchymal transition, in murine airway epithelial cells. Because Na,K‐ATPase expression is decreased in some forms of cancer and is critical for establishing cell polarity and suppressing cell motility, we analyzed S163 mutants in an epithelial cell scratch‐wound model as a measure of cell migration. Wild‐type FXYD5 overexpression increased reepithelialization (p<0.0001), which was further increased in S163D mutants (p<0.005). However, S163A mutants inhibited epithelial cell migration compared with wild‐type FXYD5 overexpression (p<0.0001). We conclude that negative charge at S163 regulates FXYD5/Na,K‐ATPase interaction and that this interaction modulates cell migration across a wound in airway epithelial cells.
In vitro multispecies Lubbock chronic wound biofilm model
Authors: Sun Yan , Dowd Scot E. , Smith Ethan , Rhoads Dan D. , Wolcott Randall D.
Published: August 2010

Multispecies biofilms are becoming increasingly recognized as the naturally occurring state in which bacteria reside. One of the primary health issues that is now recognized to be exacerbated by biofilms are chronic, nonhealing wounds such as venous leg ulcers, diabetic foot ulcers, and pressure ulcers. Arguably three of the most important species associated with multispecies biofilms that our group sees clinically are Pseudomonas aeruginosa, Enterococcus faecalis, and Staphylococcus aureus. This study was conducted to address the need for a chronic pathogenic biofilm laboratory model that allows for cooperative growth of these three organisms. We have developed a novel media formulation, simple laboratory system, quantitative polymerase chain reaction for monitoring population dynamics, and methods for objectively and subjectively measuring biofilm formation. The Lubbock chronic wound pathogenic biofilm withstood treatment with a 50‐fold higher concentration of bleach than that which was completely bacteriocidal for fully turbid planktonic cultures. The Lubbock chronic wound pathogenic biofilm when treated with biofilm effectors such as gallium nitrate and triclosan responded with selective inhibition of Pseudomonas aeruginosa or Staphylococcus aureus, respectively, as has been reported in the literature. The ability of this 24‐hour model to react as predicted using known biofilm effectors suggests that it will lend itself to future work in the development and testing of first‐generation chronic wounds pathogenic biofilm therapeutics. We have defined a realistic in vitro multispecies biofilm model simulating the functional characteristics of chronic pathogenic biofilms and developed effective tools for its characterization and analyses.
Localization of small leucine‐rich proteoglycans and transforming growth factor‐β in human oral mucosal wound healing
Authors: Honardoust Dariush , Eslami Ameneh , Larjava Hannu , Häkkinen Lari
Published: August 2010

Wound healing in oral mucosa is fast and results in little scar formation as compared with skin. The biological mechanisms underlying this property are poorly understood but may provide valuable information about the factors that promote wound regeneration. Small leucine‐rich proteoglycans (SLRPs) decorin, biglycan, fibromodulin and lumican are extracellular matrix molecules that regulate collagen fibrillogenesis, inhibit transforming growth factor‐β (TGF‐β) activity and reduce scarring. In the present study, we analyzed accumulation of SLRPs and TGF‐β during non‐scarring human oral mucosal wound healing. Biopsies were collected from healthy volunteers from unwounded tissue and from standardized experimental wounds 3–60 days postwounding. Localization of SLRPs, TGF‐β1 and TGF‐β3 was analyzed by immunohistochemical staining and quantitated by image analysis. Double immunostaining was used to study localization of SLRPs or active TGF‐β in distinct cells. Decorin, biglycan, fibromodulin, and TGF‐β isoforms showed significantly increased accumulation in the wound extracellular matrix and distinct wound cells while the abundance of lumican in the extracellular matrix was strongly reduced during wound healing. Localization and abundance of fibromodulin, lumican, and TGF‐β isoforms was also spatiotemporally regulated in the wound epithelium. The findings suggest that SLRPs regulate wound reepithelialization and connective tissue regeneration during oral mucosal wound healing.
THE FRESHWATER DIATOM CYMBELLA CURSIFORMIS NOM. NOV.1,2
Authors: Hufford Terry L. , Collins Gary B.
Published: August 2010

The morphology, ecology, and taxonomy of a pennate diatom, Cymbella cursiformis nom, nov. (family Cymbellaceae), from Cedar Bog, Champaign County, Ohio, originally described from Tasmania by Hustedt as Cymbella cistula var. gracilis, are reported. Photographs of specimens on Hustedt's type, slide, are indistinguishable from Cedar Bog specimens. Scanning electron microscopy and light microscopy reveal that, this taxon cannot be considered a variety of Cymbella cistula (Ehr.) Kirchn.; therefore it has been raised to specific rank.
A Polymorphism in GABRA2 Is Associated With the Medial Frontal Response to Alcohol Cues in an fMRI Study
Authors: Kareken David A. , Liang Tiebing , Wetherill Leah , Dzemidzic Mario , Bragulat Veronique , Cox Cari , Talavage Thomas , O’Connor Sean J. , Foroud Tatiana
Published: August 2010

Background:  Significant evidence has accumulated to suggest an association between single‐nucleotide polymorphisms (SNPs) in the GABRA2 gene and alcoholism. However, research has yet to show an association between these polymorphisms and the human brain’s reward system function. In this study, we stratified subjects who had participated in an fMRI study of alcohol cue responses according to their genotype at a SNP in GABRA2 (rs279871) shown to be associated with alcohol dependence (Edenberg et al., 2004).

Methods:  Genotyping showed 13 subjects to be homozygous for the high‐risk allele (AA), and 23 subjects to be heterozygous (AG). In fMRI, subjects were exposed to the aromas of their preferred alcoholic drink odors (AO), as well as to appetitive control odors (ApCO) under both alcohol intoxication and placebo control conditions.

Results:  Homozygous AA subjects had a larger [AO > ApCO] response than did AG subjects in medial frontal cortical areas thought to code reward value. However, AG subjects had a larger [AO > ApCO] effect in the ventral tegmental area. Alcohol intoxication did not alter these group differences.

Conclusions:  These are the first data to suggest that GABRA2 genotype could affect the brain’s responses to cues associated with alcohol.
Toddler Emotion Regulation With Mothers and Fathers: Temporal Associations Between Negative Affect and Behavioral Strategies
Authors: Ekas Naomi V. , Braungart‐Rieker Julia M. , Lickenbrock Diane M. , Zentall Shannon R. , Maxwell Scott M.
Published: August 2010

The present study investigated temporal associations between putative emotion regulation strategies and negative affect in 20‐month‐old toddlers. Toddlers’ parent‐focused, self‐distraction, and toy‐focused strategies, as well as negative affect, were rated on a second‐by‐second basis during laboratory parent–toddler interactions. Longitudinal mixed‐effects models were conducted to determine the degree to which behavioral strategy use predicts subsequent negative affect and negative affect predicts subsequent strategy use. Results with mother–toddler and father–toddler dyads indicated that parent‐focused strategies with an unresponsive parent were followed by increases in negative affect, whereas toy‐focused strategies were followed by decreases in negative affect. Results also indicated that toddler negative affect serves to regulate behavioral strategy use within both parent contexts.
Quantification of human platelet antigen‐1a antibodies with the monoclonal antibody immobilization of platelet antigens procedure
Authors: Bertrand Gérald , Jallu Vincent , Gouet Maxime , Kjaer Killie Mette , Lambin Patrick , Husebekk Anne , Kaplan Cécile
Published: August 2010

BACKGROUND: Fetal‐neonatal alloimmune thrombocytopenia results from a maternal alloimmunization against fetal platelet (PLT) antigens. In Caucasian persons, human PLT antigen‐1a (HPA‐1a) is the most frequently implicated antigen. Our aim was to develop a method supported by a mathematical approach to quantify HPA‐1a antibodies.

STUDY DESIGN AND METHODS: The monoclonal antibody immobilization of PLT antigens (MAIPA) protocol was applied on serial dilutions of a serum with high concentration of HPA‐1a antibodies used as reference. Two mathematical procedures were used: the standard curve was constructed with a linear and a logistic regression. These regressions were used to calculate the amount of antibody in 14 test serum samples from mothers with detectable anti‐HPA‐1a.

RESULTS: Similar quantifications were obtained with both mathematical procedures. In 11 serum samples, the results, expressed in arbitrary units (AU), ranged from 62 ± 4 to 1096 ± 19 with the linear regression and from 62 ± 4 to 1117 ± 38 with the logistic regression. In addition, the linear regression allowed us to measure 3 test serum samples with low antibody concentrations from 4 ± 2 to 44 ± 3 AU, whereas the logistic regression was not suitable for the quantification of antibodies in these serum samples.

CONCLUSION: A simple quantification method was developed for anti‐HPA‐1a, based on the MAIPA procedure, allowing further studies concerning correlations between anti‐HPA‐1a quantification and clinical relevance.
Possible involvement of heparin‐binding protein in transfusion‐related acute lung injury
Authors: Yasui Kazuta , Furuta Rika A. , Matsuyama Nobuki , Fukumori Yasuo , Kimura Takafumi , Tani Yoshihiko , Shibata Hirotoshi , Hirayama Fumiya
Published: August 2010

BACKGROUND: In antibody‐mediated nonhemolytic transfusion reactions, transfusion‐related acute lung injury (TRALI) tends to occur typically within 2 hours after a blood transfusion. White cell antibodies or immune complexes have been frequently shown to be associated with the syndrome, although the mechanisms by which they induce TRALI are poorly understood. The aim of this study was to characterize soluble mediators that are released from cells at an early stage after immune stimulation.

STUDY DESIGN AND METHODS: To explore the mechanism of TRALI, an in vitro whole‐blood cell culture assay was established in which cells were stimulated by human antibodies and the activation of neutrophils was monitored by a cell surface marker (Mac‐1) with flow cytometry and by measurement of the release of soluble factors, including perforin, interleukin‐6, tumor necrosis factor‐α, and heparin‐binding protein (HBP) with enzyme‐linked immunosorbent assays. In addition, the involvement of two neutrophil FcγRs (FcγRIIIb and FcγRIIa, also known as CD16 and CD32, respectively) was examined during antibody‐induced cell activation with anti‐FcγR blocking antibodies.

RESULTS: Substantial amounts of HBP were released within 30 minutes of stimulation by human antibodies, although other soluble mediators were not released within the same period. Furthermore, the release of HBP was mediated via signals through both FcγRIIIb and FcγRIIa.

CONCLUSION: HBP appears to be one of the primary effector molecules of antibody‐mediated nonhemolytic transfusion reactions including TRALI.
Prognostic Significance of Large Mediastinal Involvement in Hodgkin's Disease
Authors: Mazza P. , Lauria F. , Sciascia R. , Emiliani E. , Fiacchini M. , Baccarani M. , Frezza G. , Dominici G. , Cantore M. , Faedi M. , Gherlinzoni F. , Tura S.
Published: August 2010

Relapse rates of 75 patients with previously untreated Hodgkin's disease with stages I and II nodular sclerosis were analyzed according to the mediastinal involvement. The overall relapse rate was 22.6%. The probability of relapse was much greater for patients with large mediastinal involvement (66.6%) compared with 17% for patients with small mass, and 11.7% of patients without mediastinal involvement (P < 0.001). There was no significant difference in recurrence rates between patients without mediastinal mass and patients with a small mass, and in these patients adjuvant chemotherapy MOPP after radiotherapy showed an evident benefit in reducing the relapse rate. On the other hand, no benefical effect of adjuvant chemotherapy was observed in patients with large mediastinal involvement. Finally, in the 17 relapsing patients, ‘salvage’ chemotherapy was less effective in patients with large mediastinal mass than in those with small or no mediastinal involvement.
Iso‐immuno‐electrophoresis of plasma fibrinogen Preliminary results
Authors: KIERULF P.
Published: August 2010

Plasma (citrated) from normal single donors, plasma treated with plasmin, and plasma from two patients during different stages of disease (one pneumonia, one myocardial infarction) were subjected to isofocusing (1st dimension) in agarose (IsoGel, Marine Colloids) (1 mm, pH 3.5–9.5, Ampholine LKB) using a water‐cooled flat‐bed equipment (Multiphore LKB). Subsequent to isofocusing immuno‐profiles were established by electrophoresing focused species into fibrinogen antibody‐containing agarose (2nd dimension), and developing the plates with Coomassie Brilliant Blue. Normal plasma always demonstrated a broad, partly peaked (2–3 peaks, pI 5.5, 5.3, 5.1) iso‐immuno‐profile, suggesting normal plasma fibrinogen to consist of several charged species. Plasminized normal plasma, and patient plasma (in the absence of significant amounts of serum‐FDP) regularly demonstrated evolving iso‐immuno‐patterns, suggesting the present two‐dimensional iso‐immuno‐profiling of plasma fibrinogen to be a valuable accretion to other methods used to demonstrate molecular heterogeneity.
Mechanisms of Decreased Leucocyte Adhesiveness and Migration in Plasma from Patients with IgG Myelomatosis
Authors: Hopen G. , Glette J. , Matre R.
Published: August 2010

The adhesiveness of leucocytes in human plasma was decreased by the addition of IgG M‐protein, commercial IgG, F(ab)2‐fragments, or bovine serum albumin (BSA). Leucocyte migration was inhibited by IgG M‐protein and commercial IgG, but not by reduced and alkylated IgG, F(ab)2‐fragments or BSA. Guinea pig serum (GPS) strongly enhanced leucocyte migration in Hanks' balanced salt solution. This effect was abolished by hydrazine treatment or heating, and reduced, but not abolished, by zymosan treatment. C4‐deficient guinea pig serum (C4‐def‐GPS), but not zymosan treated C4‐def‐GPS stimulated migration. In IgG myeloma sera migration was enhanced by the addition of C4‐def‐GPS and by untreated or heated, but not hydrazine treated GPS, and inhibited by zymosan treated GPS. IgG M‐protein inhibits leucocyte adhesiveness and migration. Migration is specifically inhibited by the intact IgG, probably through complement activation with consumption of heat stable chemokinetic factor(s) and production of migration inhibitors.
Sickle cell β‐thalassaemia compared with sickle cell anaemia in Algeria
Authors: Belhani M. , Morle L. , Godet J. , Bachir D. , Henni T. , Zerhouni F. , Bensenouci A. , Colonna P.
Published: August 2010

Clinical, haematological and biochemical features in 42 subjects with S‐beta thalassaemia (31 subjects with S‐beta° thalassaemia and 11 subjects with S‐beta+ thalassaemia); and in 42 with homozygous sickle cell disease were compared. Persistent splenomegaly was more common and painful crises less common in the S‐beta thalassaemia group. Total Hb was higher and reticulocyte count lower in S‐beta+ thalassaemia than in S‐beta° thalassaemia or SS disease. Microcytosis was marked in the S‐beta thalassaemia group while the MCV was normal in sickle cell anaemia. Hb F was significantly higher in the S‐beta° thalassaemia group, without any influence on the severity of the disease. Many features suggest that sickle cell thalassaemia is more severe in Algeria than in Negro subjects and similar to the disease in Italian patients.
Clinical course and survival in 16 patients with localized plasmacytoma
Authors: Wollersheim H. C. H. , Holdrinet R. S. G. , Haanen C.
Published: August 2010

The clinical course of 16 patients with localized plasmacytoma, 9 with solitary plasmacytoma of bone (SPB), and 7 with extramedullary plasmacytoma (EMP) are presented. Median follow up of SPB was 77 months and of EMP 74 months. The EMPs were localized in the upper respiratory tract (4 cases), the gastro‐intestinal tract (2 cases) and the skin (1 case). The SPBs were localized in the ribs (3 cases), the spine (3 cases), the humoral bone (1 case), the skull (1 case) and the mandibular bone (1 case). In 3 patients with EMP and in 5 patients with SPB, the monoclonal protein could be determined at presentation. Local recurrences following surgical removal or irradiation occurred in 2 patients with EMP and in 1 patient with SPB after 12–60 months. Classical multiple myeloma developed in 3 patients with EMP and in 4 patients with SPB, 9–130 months after diagnosis. The monoclonal protein level proved to be a useful disease marker which (re)appeared at local recurrence or at dissemination. The median survival of the 16 patients with solitary plasmacytoma was 138 months, which is considerably longer than the median survival of 42 months in 14 stage‐I patients with classical multiple myeloma who were observed during the same follow‐up period.
A stage classification for prognosis in adult acute myelogenous leukaemia based upon patients' age, bone marrow karyotype and clinical features
Authors: Bernard Philippe , Reiffers Josy , Lacombe Francis , Dachary Dominique , Boisseau Michel R. , Broustet Antoine
Published: August 2010

In 71 adult acute myelogenous leukaemia (AML) cases, the relationship between well‐known prognostic features and complete remission (CR) rate and survival was studied. These features were: (i) bone marrow karyotype classified NN, AN, AA according to Sakurai & Sandberg (5); (ii) patients' age; (iii) clinical ‘negative prognostic features' (NPF): previous history of preleukaemia, septicaemia or pneumonia, hyperleucocytosis, associated pathology (diabetes, obesity, renal insufficiency etc.). 59 years of age was found to be a frontier between 2 homogenous groups having quite different prognosis. The NN/AN/AA classification had good prognostic value (CR rate and survival) in patients under 59 years, but not in older patients. In those patients over 59, a significant difference in CR rate and survival appeared between cases with NPF and those without.

For each feature having an established relationship to survival, a panel of prognostic points was determined as follows: age over 59 (1 point), AA karyotype in patients under 59 (2 points), NPF in patients over 59 (1 point). Using this stage classification, it was possible to classify every case into 1 of 3 groups (i.e.: 0 points, 1 point, 2 points). The life‐table analysis of these 3 groups showed very significant differences. The median survival times were 18.5 months, 5.2 months and 1.3 month, for the 0‐point group (26 cases), the 1‐point group (19 cases) and the 2‐point group (26 cases), respectively.
Ultrastructural localization of acid phosphatase in immunologically defined neoplastic lymphocytic cells and hairy cells A comparison between two different substrates
Authors: BOESEN ANNE MARIE
Published: August 2010

The subcellular localization of acid phosphatase (AcP) in various immunologically‐defined neoplastic lymphoid cells including hairy cells was investigated by electron microscopy. 2 substrates, naphthol‐AS‐BI‐phosphoric acid (naphthol‐AS‐BI‐P) and sodium β‐glycerophosphate, were compared. By incubation in naphthol‐AS‐BI‐P containing medium, the reaction product was found located in granules, vesicles, the Golgi apparatus, the rough ER including the nuclear envelope in the cells of T ALL, T CLL and HCL. A typical pattern of reaction was observed for each of these disorders: enzyme‐positive Golgi membranes and neighbouring granules, clustered in the nuclear notch in T cell‐derived lymphoblasts; enzyme‐positive granules around Gall bodies, aggregated paranuclearly in T CLL lymphocytes and enzyme‐positive scattered cytoplasmic granules and vesicles in hairy cells. Enzyme activity was occasionally seen in singly‐occurring granules in the cells of cALL, B CLL and B PLL, rarely in other substructures. With the Gomori method using β‐glycerophosphate as substrate, the enzyme reaction was limited primarily to lysosomal sites and was seldom observed in other organelles. Tartrate‐resistant AcP was found in the majority of hairy cells and in a few prolymphocytes, located in the same structures as AcP without tartrate.
Effectiveness of Pentostatin (2‘‐Deoxycoformycin) in Refractory Lymphoid Neoplasms
Authors: Spiers A. S. D. , Ruckdeschel J. C. , Horton J.
Published: August 2010

Pentostatin (2′‐deoxycoformycin, DCF) was administered to 17 patients with a variety of lymphoid neoplasms, both T‐ and B‐cell, that were refractory to conventional treatments. Several responses and 2 complete remissions occurred. Toxic effects were less severe than previously described: this may be attributable to relatively low doses of DCF or to precautions taken to prevent tumour lysis syndrome. DCF appears valuable as a second‐line treatment in non‐Hodgkin's lymphomas and as initial treatment in T‐cell chronic lymphocytic leukaemia and mycosis fungoides. Although myelosuppression is mild, immunosuppression and superinfection are potential hazards of treatment with DCF. The ocular toxicity of DCF, previously described as conjunctivitis, appears to be a keratitis of moderate severity which requires further study.
Prognostic factors for non‐cleaved follicular center‐cell lymphomas and immunoblastic sarcoma A Bayesian approach
Authors: AINE R. , KATAJA M. , ALAVAIKKO M.
Published: August 2010

The Bayesian multivariate statistical method was applied to determine the relative strength and optimal combination of 17 variables in predicting survival of 151 patients with non‐Hodgkin's lymphomas assigned as non‐cleaved follicular center‐cell and immunoblastic sarcoma types according to the classification of Lukes & Collins. Considering all the factors simultaneously, the analysis showed that the combination of stage, Hb level and location of the lymphoma was included in the best predictive model at each survival time studied. Additional factors were erythrocyte sedimentation rate, thrombocyte count and leucocyte count. Of the histological variables, only growth pattern and mitotic ratio in the biopsy specimen remained significant. At manually controlled computer simulation with these best indicators, this model would have given a correct classification for 69–78 % of the patients at the 4 survival times studied. One can thus expect about 70 % correct prognoses using this model.
The in vitro growth pattern of human bone marrow in methylcellulose stimulated by different concentrations of conditioned medium
Authors: HAAK H. L. , GOSELINK H. M. , VEENHOF W. F. J. , BLOTKAMP J. , JANSEN J.
Published: August 2010

The proliferation of human bone marrow in methylcellulose stimulated by various concentrations of conditioned medium (CM) and observed at various intervals was studied. The growth kinetics of granulocytic aggregates was found to differ from monocytic clusters and colonies. Granulocytic aggregates showed a consistent and reproducible dose‐response relationship; at day 7, the maximum number of granulocytic aggregates was found at 4% CM. At higher levels, the total number of aggregates decreased, while the number of cells per aggregate increased. The number of macrophage aggregates was far less, depending on the CM concentration. Photographic interval studies showed that at high concentrations of CM, clusters and colonies were formed earlier, but also coalesce to form one colony. Our results suggest that the proliferation kinetics of granulocytic aggregates are complex and preclude simple statements about sensitivity to colony‐stimulating activity.
The great majority of childhood lymphoblastic leukaemias are identified by monoclonal antibodies as neoplasias of the B‐cell progenitor compartment
Authors: Melsom Haakon , Funderud Steinar , Lie Sverre O. , Godal Tore
Published: August 2010

36 acute leukaemias in children, 24 lymphoblastic and 14 myelogenic, have been examined with a set of 10 monoclonal antibodies by indirect fluorescent staining. In the lymphoblastic group the leukaemic cells of 4 children were found to have T‐cell phenotype, while 19 of the other 20 T‐cell phenotype negative cases were found to be positive for the c‐ALL antigen. All 20 were negative for surface immunoglobulin as well as cytoplasmic μ‐heavy chains. However, 17 (85 %) reacted positively with the monoclonal antibody AB‐1 which we have developed against a B‐cell lymphoma, thus revealing B‐lineage specificity. Another B‐lineage‐associated antibody (AB‐2) reacted with 8/20 (40 %) of the cases with distribution similar to B‐1. These findings suggest that the great majority of non‐T‐non‐B acute lymphoblastic leukaemias are neoplasms derived from the B‐cell progenitor compartment. Moreover, monoclonal antibody testing allows further sub‐categorization in this group. Similarly the acute‐myelogenic leukaemia group could be subdivided into phenotypic subsets. The importance of using panels of monoclonal antibodies in the diagnosis of acute leukaemias is discussed.
Platelet glyoxalases in thrombocytosis
Authors: Leoncini G. , Maresca M. , Balestrero F. , Armani U. , Piana A.
Published: August 2010

In platelets of patients suffering from thrombocytosis due to myeloproliferative disorders, glyoxalase I activity is significatively higher than in controls (P < 0.01), while glyoxalase II levels are the same (P < 0.3). The endocellular concentration of glutathione is also increased in patients. Km values for methylglyoxal (glyoxalase I) and S‐lactoylglutathione (glyoxalase II) are identical both in normal and pathological subjects, as are the thermostability of the two enzymes. The higher activity observed for glyoxalase I in patients could be related to a specific role of this enzyme in platelets.
Kinetic evaluation of four Factor VIII concentrates by model‐independent methods
Authors: Matucci M. , Messori A. , Donati‐Cori G. , Longo G. , Vannini S. , Morfini M. , Tendi E. , Rossi‐Ferrini P. L.
Published: August 2010

Single‐dose kinetics of 4 Factor VIII concentrates (Kryobulin, Hemofil T, Koate, cryoprecipitate) were studied in 41 patients with haemophilia‐A. Model‐independent methods were adopted for calculating the kinetic parameters (area under the curve, clearance, area under the moment curve, mean residence time, volume of distribution at steady‐state). No substantial difference was observed in the kinetic characteristics of the 4 Factor VIII concentrates. A considerable interindividual variability of the calculated kinetic parameters was demonstrated for all concentrates. Our findings support the need to individualize Factor VIII dosage.
Platelet 5‐Hydroxytryptamine transport and storage in myeloproliferative disorders
Authors: Cortellazzo Sergio , Viero Piera , Buczko Wlodzimierz , Barbui Tiziano , de Gaetano Giovanni
Published: August 2010

Platelets of patients with myeloproliferative disorders (MD) such as polycythaemia vera (PV), chronic myelogenous leukaemia (CML), idiopathic myelofibrosis (IM) and essential thrombocythaemia (ET) have been found to have low 5HT levels measured both by a fluorimetric and a liquid chromatographic assay. Km and Vmax for platelet active uptake of 3H‐5HT were not significantly different in controls and patients. Inhibition of 5HT reuptake by imipramine or induction of moderate release by fenfluramine were not sufficient to distinguish the group of MD platelets from controls, although some patients had less of a tendency to retain intraplatelet amine. The low platelet 5HT content found in our patients seems not to be the consequence of disturbed active transport of 5HT across platelet membrane. Although defective storage of this amine within the cell is probable, the results of the present study do not rule out the possibility that platelets from MD patients undergo in vivo activation by endogenous stimuli not inhibited by aspirin. 10 d treatment with aspirin did not result in any significant rise in intraplatelet 5HT concentration.
T‐cell subsets in malignant lymphomas and monoclonal gammopathies
Authors: Bergmann Lothar , Mitrou Paris S. , Kelker Wolfgang , Weber Kai C.
Published: August 2010

159 patients with malignant lymphomas or monoclonal gammopathies were investigated for lymphocytes and their subsets using conventional surface markers and a panel of monoclonal antibodies. In untreated patients with Hodgkin's disease, non‐Hodgkin lymphomas and multiple myeloma, (MM) a reduction of T‐cells and especially of the “helper/inducer” subset (OKT4+) was found to be a common phenomenon. The major abnormalities occurred in advanced stages of disease. Patients previously treated by chemo‐ and/or radiotherapy had a further decrease of T‐cells, whereas the loss of OKT4+ cells was more pronounced than that of the “suppressor/cytotoxic” lymphocytes (OKT8+).

The alterations of lymphocyte subsets persisted even in long‐term remitters. Comparing the lymphocyte subsets in MM and benign monoclonal gammopathies (BMG), patients with BMG showed a significant reduction in OKT8+ cells, whereas the OKT4+ population was within normal range, resulting in a significant elevation of the OKT4/OKT8‐ratio compared to the controls and untreated multiple myeloma.
Iron chelation using subcutaneous infusions of diethylene triamine penta‐acetic acid (DTPA)
Authors: Pippard M. J. , Jackson M. J. , Hoffman K. , Petrou M. , Modell C. B.
Published: August 2010

The iron chelating ability and potential toxicity of subcutaneous infusions of the calcium and zinc salts of diethylene triamine penta‐acetic acid (DTPA) have been assessed in metabolic balance studies in 2 iron‐loaded thalassaemic patients. Infusions of calcium DTPA were locally well tolerated and the drug was as effective as desferrioxamine in mobilising iron. However, daily infusions in the 1st patient also produced symptomatic zinc depletion which could not be controlled by simultaneous oral zinc supplements. Zinc DTPA proved ineffective as an iron chelator, but zinc balance could be maintained in the 2nd patient by combining intermittent (every 4 d) use of calcium DTPA with oral zinc supplements. Combined studies with desferrioxamine and calcium DTPA showed the drugs to have additive effects, probably as a result of the chelation of iron from different body sites.
Mechanistic study of endogenous skin lesions in diabetic rats
Authors: Chen Xiang‐fang , Lin Wei‐dong , Lu Shu‐liang , Xie Ting , Ge Kui , Shi Yong‐quan , Zou Jun‐jie , Liu Zhi‐min , Liao Wan‐qing
Published: August 2010

Please cite this paper as: Mechanistic study of endogenous skin lesions in diabetic rats. Experimental Dermatology 2010; 19: 1088–1095.

Abstract:  Pathological and physiological changes in dermal tissue in a rat model of diabetes mellitus (DM) were investigated. Sixteen male 8‐week‐old Sprague–Dawley rats were randomized into two groups of eight, the DM group (Group DM) and the normal control group (Group (NC) normal control). Group DM rats were injected with streptozotocin (STZ) intraperitoneally at a dose of 65 mg/kg body weight. Group NC rats were injected with the same volume of citric acid buffer. All rats were sacrificed 12 weeks later. The impact of exposure to (AGE) advanced glycation end products‐modified human serum albumin (AGE‐HSA) on epidermal cells and ECV304 cells was evaluated in cell culture experiments. The diabetic rats exhibited changes in skin tissue, including a decrease in thickness, disappearance of the multilayer epithelium structure, degeneration of collagen fibres and an increase in the infiltration of inflammatory cells, in addition to a significant increase in skin glucose and AGEs. Moreover, diabetic rats had increased plasma glycosylated protein (GSP) and malondialdehyde (MDA) and decreased plasma glutathione (GSH). The percentage of epidermal cells in S phase was similar between the two group rats; however, there was a marked decrease in the G2/M phase in Group DM. Additionally, exposure of ECV304 cells to AGE‐HSA led to a time‐dependent and dose‐dependent increase in apoptosis. Therefore, the high glucose in the skin tissue, coupled with the accumulation of toxic substances such as AGEs, promote the dysfunction of dermal cells and/or the matrix. This may be a significant mechanism of diabetes‐induced early‐stage endogenous skin damage.
Amelogenesis imperfecta—towards a new classification
Authors: Aldred MJ , Crawford PJM
Published: August 2010

This editorial reviews the history of the classification of amelogenesis imperfecta (Al). The limitations of the existing classification systems are discussed. An alternative classification is proposed based upon the molecular defect, biochemical result, mode of inheritance and phenotype in the family involved. While not all of the criteria for the proposed classification can yet be addressed, this scheme is proposed for future classification of At cases and families.
Analysis of cultivable Porphyromonas gingivalis with trypsin‐like protease enzyme activity and serum antibodies in chronic adult periodontitis
Authors: Smith AJ , Wade WG , Greenman J , Addy M
Published: August 2010

OBJECTIVE: Trypsin‐like protease (TLPase) enzyme produced by Porphyromonas gingivalis has been implicated as a virulence factor in the pathogenesis of periodontal disease. The aims of this study were to investigate the relationship between cultivable P. gingivalis, TLPase enzyme activity (BANA hydrolysis) and serum antibody levels against cell sonicate and a purified TLPase antigen from P. gingivalis W50.

MATERIALS AND METHODS: Sub‐gingival plaque samples were cultured for levels of P. gingivalis together with a chairside analysis of TLPase enzyme activity (Perioscan) from periodontitis and gingivitis sites of adult periodontitis patients. A TLPase from P. gingivalis was purified by gel filtration and ion exchange chromatogra‐phy from the vesicle fraction for use as a test antigen. RESULTS: Elevated levels of P. gingivalis were found at periodontitis sites, however, there was no correlation with sub‐gingival plaque TLPase enzyme activity. Adult periodontitis patients had higher levels of IgG and IgA against cell sonicate and TLPase antigens than did controls. Those patients who were P. gingivalis culture‐positive demonstrated an elevated immune response against both cell sonicate and TLPase when compared to P. gingivalis culture‐negative patients. Treatment resulted in an improvement of clinical indices and no cultivable P. gingivalis could be recovered from the treated sites and there was a concomitant decrease in IgG levels against the TLPase. There was no significant difference in BANA hydrolysis at gingivitis sites or periodontitis sites after treatment.

CONCLUSIONS Further longitudinal studies are suggested to investigate the role of the TLPase in the response to treatment of chronic adult periodontitis patients.
Cytokines and cell signalling in the periodontium
Authors: Hughes FJ
Published: August 2010

A large number of potential regulatory mechanisms have been described which may be involved in the control of cell function in the periodontium. In this review, soluble effector molecules which may regulate normal cell turnover and which may control the maintenance of the per‐iodontal space are considered. There is evidence for the involvement of growth factors including EGF, PDGF, FGFs, IGF I & II and TGFβ in these processes. The role of bone morphogenetic proteins (BMPs) in periodontal turnover is of considerable interest as they appear to be able to regulate all stages of this process from specifying cell commitment to regulating differentiated cell function.

Empirical evidence suggests the importance of mechanical stimulation in controlling the maintenance of the periodontal ligament space. The wide range of effects of mechanical stimulation are briefly reviewed and the central role of prostaglandins is considered. Recent evidence suggests the involvement of nitric oxide (NO) in the regulation of mineralised tissue function, and the potential role of NO in maintenance of the ligament space is considered. Further studies are required which address the interactions between all of these mechanisms in order to determine the key factors which may control periodontal cell function. For the future an understanding of these interactions has the potential to lead to important clinical developments in periodontal and orthodontic therapy.
Human gingival keratinocytes express E‐selectin (CD62E)
Authors: Pietrzak ER , Savage NW , Walsh LJ
Published: August 2010

OBJECTIVES: E‐selectin (CD62E) is an adhesion molecule that participates in the binding of leukocytes to activated blood vascular endothelium. The present study was undertaken to characterise the pattern of E‐selectin expression on epithelial cells of the gingival crevice and oral epithelium.

METHODS: A panel of six anti‐E‐selectin monoclonal antibodies was reacted with cryosections of human gin‐giva and cytospins of cultured gingival keratinocytes.

RESULTS Three antibodies raised against leukocyte binding epitopes of the molecule (H4/18, HI8/7, I.2B6) were reactive with gingival keratinocytes, three were negative (4DI0, BBAl, BBA2), while all six reacted with endothelial cells. Staining with H18/7 and 1.2B6 was weaker and more variable than with H4/18. In cell culture, levels of E‐selectin expression reduced slowly, and were only modestly increased by treatment with exogenous TNFα, a known inducer of E‐selectin on endothelium.

CONCLUSIONS: E‐selectin epitopes are expressed on gingival keratinocytes. These binding epitopes may be involved in the traffic of leukocytes in this tissue compartment.
Diagnostic strategies of periodontitis based on the molecular mechanisms of periodontal tissue destruction
Authors: Okada H , Murakami S , Kitamura M , Nozaki T , Kusumoto Y , Hirano H , Shimauchi H , Shimabukuro Y , Saho T
Published: August 2010

OBJECTIVE: Periodontitis is a disease showing differences in disease progression between patients and between sites within a patient. Routine clinical examinations today are not useful enough to distinguish susceptible patients and active lesions from resistant patients and chronic lesions. Diagnostic markers should be pathogenic and inflammatory factors participating in periodontal tissue destruction. These are both local and systemic factors.

MATERIALS AND METHODS: First of all, pathogenic factors and proinflammatory cytokines or mediators in gingival crevicular fluid (GCF) were examined and the difference was found between active and inactive periodontitis lesions distinguished by attachment loss. Active lesions were detected by discriminant‐function analysis of these examinations, although the sensitivity of differential diagnosis was low. Then, we established a novel needle biopsy for understanding the pathophysiological conditions elicited in active and chronic inflammatory processes of periodontal tissue destruction. A variety of cytokines and mediators were detected in biopsied specimens by reversed transcription polymerase chain reactions (RT‐PCR). Cytokine profiles were varied in inflamed periodontal biopsies. As IFNy mRNA expression was enhanced in inflamed gingiva, antigen‐presenting‐cell (APC) functions of human gingival fibro‐blasts (HGF) were examined.

RESULTS: Despite the phenotypical resemb!ance of IFNγ‐treated HGF to so‐called APC, HLA‐DR positive HGF could not induce proliferation but suppressed proliferation of alloreactive peripheral blood T cells (PBT). However, HLA‐DR positive HGF stimulated the prolifer‐ative responses of PBT which had been primed with allo‐APC. Regulatory immune responses by IFNγ were different in T cell conditions.

CONCLUSIONS Various kinds of cytokines participated in periodontal inflammation, and every cytokine is multifunctional. Complex and compound inflammatory processes can be clarified by examining cytokine networks and the precise effects of each cytokine on each of the cell types comprising periodontal tissue. It is, therefore, necessary for establishing diagnostic strategies to integrate pathogenic and inflammatory factors in periodontal tissue destruction.
Dietary factors in oral leukoplakia and submucous fibrosis in a population‐based case control study in Gujarat, India
Authors: Gupta PC , Hebert JR , Bhonsle RB , Sinor PN , Mehta H , Mehta FS
Published: August 2010

OBJECTIVES: To investigate the relationship of specific nutrients and food items with oral precancerous lesions among tobacco users.

DESIGN: A population‐based case‐control study. SETTING: Villages in Palitana taluk of Bhavnagar district, Gujarat, India.

SUBJECTS AND METHODS: An interviewer‐administered food frequency questionnaire, developed and validated for this population, was used to estimate nutrient intake in blinded, house‐to‐house interviews. Among 5018 male tobacco users, 318 were diagnosed as cases. An equal number of controls matched on age (±5 years), sex, village, and use of tobacco were selected.

MAIN OUTCOME MEASURES: Odds ratios (OR) from multiple logistic regression analysis controlling for relevant variables (type of tobacco use and economic status).

RESULTS: A protective effect of fibre was observed for both oral submucous fibrosis (OSF) and leukoplakia, with 10% reduction in risk per g day‐1 (P < 0.05). Ascorbic acid appeared to be protective against leukoplakia with the halving of risk in the two highest quartiles of intake (versus the lowest quartile: OR = 0.46 and 0.44, respectively; P < 0.10). A protective effect of tomato consumption was observed in leukoplakia and a suggestion of a protective effect of wheat in OSF.

CONCLUSION: In addition to tobacco use, intake of specific nutrients may have a role in the development of oral precancerous lesions.