bluuebunny/crossref_metadata_2025 huggingface dataset:
Thrombocytopenic purpura and cardiomyopathy in pregnancy reversed by combined plasma exchange and infusion
Authors: Cosmai Elizabeth M. , Puzis Leopold , Tsai H.‐M. , Lian Eric C.‐Y.
Published: March 2003

Abstract: Thrombocytopenia and hemolytic anemia have been seen with thrombotic thrombocytopenic purpura (TTP), HELLP syndrome (hemolysis, elevated liver enzymes, and low platelets), and hemolytic uremic syndrome (HUS). Differentiating between TTP, HUS, and HELLP syndrome is often difficult. Coexistence of TTP and HELLP is possible. Cardiomyopathy occurring in pregnancy can be idiopathic or associated with TTP. We describe a previously healthy woman who developed thrombocytopenia and hemolysis at 34 wk gestation. The patient underwent delivery after transfusion with platelets and RBCs. The suspicion of TTP was raised, and plasma exchange was begun by the third hospital day. On the seventh day of treatment, the patient developed shortness of breath, and an echocardiogram showed global hypokinesis with an ejection fraction of 25%. Plasma infusion, one unit q 4 h, was initiated in addition to the daily plasma exchange. The patient improved and her ejection fraction normalized. Plasma exchange and infusion and corticosteroids were gradually tapered off. von Willebrand factor (vWF) protease activity in the plasma upon transfer was completely deficient with the presence of inhibitor. This case illustrates that vWF protease assay and detection of inhibitor can be used for the diagnosis of TTP during pregnancy; and a severe cardiomyopathy in TTP can be reversed rapidly with combined plasma exchange and infusion.
De novo acute myeloid leukemia with multilineage dysplasia: treatment results and prognostic evaluation from a series of 44 patients treated with fludarabine, cytarabine and G‐CSF (FLAG)
Authors: Ferrara Felicetto , Palmieri Salvatore , Pocali Barbara , Pollio Filiberto , Viola Assunta , Annunziata Silvana , Sebastio Lucia , Schiavone Ettore Mariano , Mele Giuseppina , Gianfaldoni Giacomo , Leoni Franco
Published: March 2003

Abstract: Objectives
: To evaluate therapeutic results and prognostic factors from a series of 44 patients affected by
de novo
acute myeloid leukemia with multilineage dysplasia (MD‐AML), treated with the combination of fludarabine, cytarabine and G‐CSF (FLAG).
Methods
: Forty‐four patients with
de novo
MD‐AML were treated with the FLAG regimen. The median age was 61 yr (range 31–75 yr). Induction therapy consisted of the FLAG regimen; consolidation included idarubicin plus cytarabine. Patients with a compatible donor and aged less than 55 yr were programmed to receive allogeneic bone marrow transplantation (BMT), while in those without a donor and aged less than 65 yr autologous transplantation with peripheral blood stem cells mobilized by a consolidation regimen plus G‐CSF was planned. Bone marrow harvest was performed in poor mobilizers.
Results
: Complete remission (CR) was achieved in 28 out of 44 patients (64%). Death in induction occurred in four patients (9%), while 12 patients (27%) were resistant to FLAG. Toxicity of consolidation was negligible. Most patients aged less than 60 yr and achieving CR were eligible for transplantation procedures, the main reason of exclusion being early relapse. Median overall survival and disease free survival were 16 and 22 months, respectively. Unfavorable cytogenetics was the only parameter significantly related to inferior clinical outcome following multivariate analysis.
Conclusion
: Multilineage dysplasia
per se is not an adverse prognostic factor in AML patients treated with the FLAG regimen. Favorable results are obtained in patients with intermediate karyotype, while in those with adverse cytogenetics new approaches are clearly needed. The toxicity of the regimen is also acceptable in the elderly, and following induction/consolidation, most patients may be submitted to transplantation procedures.
Endogenous endophthalmitis following disseminated fungemia due to Fusarium solani in a patient with acute myeloid leukemia
Authors: Tiribelli Mario , Zaja Francesco , Filì Carla , Michelutti Teresa , Prosdocimo Simonetta , Candoni Anna , Fanin Renato
Published: March 2003

Abstract: We report the case of a young man with a resistant acute myeloid leukemia (AML) who developed a disseminated fungemia due to Fusarium solani involving the skin and lungs, during the neutropenic phase following a chemotherapy course. Despite continuous therapy with liposomal amphotericin B, he developed a bilateral endophthalmitis that rapidly evolved to complete blindness. The patient underwent two procedures of vitrectomy, with detection of F. solani in the vitreous fluid, and continued antifungal therapy, without any recovery of visual acuity. When he eventually died due to recurrence of leukemia and hemorrhagic shock, autopsy revealed a diffuse fusarial involvement of the central nervous sysytem.
Characterization of five marker levels of the hemostatic system and endothelial status in normotensive pregnancy and pre‐eclampsia
Authors: Hayashi Masatoshi , Inoue Teruo , Hoshimoto Kazunori , Negishi Hideaki , Ohkura Takeyoshi , Inaba Noriyuki
Published: March 2003

Abstract:  Objectives
: Pre‐eclampsia is associated with changes in the hemostatic system and endothelial status. Urinary 11‐dehydrothromboxane B
2

/creatinine (11‐DTXB
2

/Cr) is a marker for platelet activation and vascular constriction, thrombin–antithrombin complex (TAT) for thrombin formation, serum thrombomodulin (TM) for endothelial damage, and β‐thromboglobulin
(β‐TG) and platelet factor 4 (PF‐4) for platelet activation and releasing reaction. The present study attempted to evaluate these five markers in normotensive pregnancy and pre‐eclampsia. Methods: These five markers were simultaneously measured in urine and blood samples from 25 women who were not pregnant (group 1, controls), 31 women with normotensive pregnancy (group 2, second controls), 22 women with mild pre‐eclampsia (group 3), and 21 women with severe pre‐eclampsia (group 4). The average gestational age was 36 wk. Results: The 11‐DTXB2/Cr, TAT, and β‐TG levels were significantly higher (P < 0.01) in groups 2, 3, and 4 than in group 1. The TM and β‐TG levels were significantly higher (P < 0.05) in group 3 than in group 2. The TM, β‐TG, and PF‐4 levels were increased significantly (P < 0.05–0.01) in group 4 compared to those in groups 1, 2, and 3. Conclusions: Platelet aggregation, vascular constriction, and thrombin formation (detected by 11‐DTXB2/Cr and TAT) may be markedly enhanced even in group 2, but further enhancement may be relatively slight in groups 3 and 4. In contrast, endothelial damage (determined by TM) and platelet release of PF‐4 may not increase significantly in group 2, but they may increase in group 4. Platelet‐release of β‐TG may be enhanced in groups 2, 3, and 4. Endothelial damage and platelet‐releasing reaction (detected by PF‐4 and β‐TG) may be significantly more enhanced in group 4 than in group 3.
Acute megakaryocytic leukemia in essential thrombocythemia: an unusual evolution?
Authors: Radaelli Franca , Mazza Rita , Curioni Elisabetta , Ciani Alberto , Pomati Mauro , Maiolo Anna‐Teresa
Published: March 2003

Abstract: Essential thrombocythemia (ET) is a chronic myeloproliferative disorder that can rarely undergo leukemic transformation either in treated (3–7%) or untreated patients (1%). Evolution to myeloblastic or myelomonoblastic acute leukemia is commonly described in the literature, whereas lymphatic and megakaryocytic forms are considered unusual. Here, we report three cases of acute megakaryocytic leukemia (LMA‐M7) among 11 acute leukemic transformations observed in our series of 321 ET patients. LMA‐M7 was diagnosed employing immunophenotyping according to FAB criteria. These recurrences of LMA M7 suggest that this kind of evolution cannot be considered rare or casual in ET.
Acquired hemophilia: a single‐center survey with emphasis on immunotherapy and treatment‐related side‐effects
Authors: Delgado Julio , Villar Ana , Jimenez‐Yuste Victor , Gago Jose , Quintana Manuel , Hernandez‐Navarro Fernando
Published: March 2003

Abstract:Objectives:Acquired hemophilia is a rare disease caused by the development of autoantibodies against factor VIII. Since 1981 we have observed 17 patients with this disorder in our institution. The objective of this survey was to assess the epidemiological features, clinical course, and mortality rate of these patients, with special emphasis on therapy‐related side‐effects. Also, we present our results with an immunosuppressive approach based on the severity of bleeding episodes.Methods: Clinical records of all patients with acquired hemophilia due to factor VIII inhibitor admitted or referred to our hospital between 1981 and 2001 were reviewed retrospectively. We collected each patient's sex, age, medical history, presenting symptoms, activated partial thromboplastin time, factor VIII activity, and inhibitor titre. Patient's clinical courses, including their bleeding episodes, response to therapy, and therapy‐related side‐effects, were also recorded.Results: Complete and partial responses were achieved in 14 and one patient, respectively (overall response rate 88%) after a median time to complete response of 3.5 months (range 30 d − 25 months). The inhibitor‐related and overall mortality rates were 12% and 29%, respectively. Side‐effects were frequent: two patients had blood‐borne infections, three patients had thrombotic complications, and nine patients had immunosuppressive therapy‐related side‐effects. In five patients, discontinuation of cyclophosphamide or prednisone was required.Conclusions: Although our response rates were remarkable, this survey showed that treatment‐related morbidity could also be very important. Therefore, it is pertinent to bear in mind these potential side‐effects in order to decide the most appropriate therapy for each particular patient.
Retinoid‐induced apoptosis in B‐cell chronic lymphocytic leukaemia cells is mediated through caspase‐3 activation and is independent of p53, the retinoic acid receptor, and differentiation
Authors: Pepper Chris , Ali Khalida , Thomas Alun , Hoy Terry , Fegan Chris , Chowdary Pratima , Kell Jonathan , Bentley Paul
Published: March 2003

Abstract: The aim of this study was to investigate the effects of all‐trans retinoic acid (ATRA) on apoptosis induction, Bcl‐2 family protein expression, and differentiation in B‐cell chronic lymphocytic leukaemia (B‐CLL) cells. ATRA induced apoptosis in all the B‐CLL samples tested, and this was accompanied by a specific reduction in Bcl‐2 and Mcl‐1 protein expression in the apoptotic cells. In contrast, Bax, p21, and p53 expression was not altered in either the viable or apoptotic B‐CLL cells, inferring that ATRA utilises a p53‐independent cell death pathway. Caspase‐3 activation was shown to be a prerequisite for ATRA‐induced apoptosis, which was inhibited by the pan‐caspase inhibitor Z‐VAD‐FMK and the caspase‐9 inhibitor Z‐LEHD‐FMK. In addition, the retinoic acid receptor (RAR) antagonist AGN194310 failed to abrogate the apoptotic effects of ATRA, indicating that RAR binding was not necessary for ATRA‐induced apoptosis. Furthermore, there was no evidence of ATRA‐induced differentiation of the B‐CLL cells in this study either in terms of altered morphology or immunophenotype. In summary these data indicate that ATRA induces apoptosis via the intrinsic apoptotic pathway, and this is independent of RAR binding, p53 activation, and cellular differentiation in B‐CLL cells.
Autologous stem‐cell transplantation for patients with acute myeloid leukemia aged over 60 yr
Authors: Ferrara Felicetto , Venditti Adriano , Carellajr Angelo Michele , Cantore Nicola , Buccisano Francesco , Tamburini Anna , Palmieri Salvatore , Mele Giuseppina , Annunziata Mario , Greco Michele Mario , Amadori Sergio
Published: March 2003

Abstract: Objectives: Preliminary reports have suggested that autologous stem‐cell transplantation (ASCT) is feasible in elderly patients with acute myeloid leukemia (AML). The objective of this study was to describe the disease characteristics and treatment results from a series of 22 elderly AML patients undergoing ASCT. Methods: The median age was 64 yr (range 61–71). Twenty patients were in first complete remission (CR1), two in CR2, and all were in performance status 0–1. The median interval between CR achievement and ACST was 3 months (range 2–5). In 20 cases peripheral blood stem cells were infused, in two bone marrow. Results: All patients had a successful engrafment. One patient (5%) died from transplant‐related complications. The median number of days to granulocytes > 500 mm−3 and platelets > 20 000 mm−3 was 11(range 9–15) and 13 (range 9–20), respectively. Non‐hematologic toxicity included WHO grade III‐IV stomatitis in 32% patients and grade IV nausea and vomiting in one (4.5%). Seven patients had fever of unknown origin, while in 14 a documented infection was diagnosed. Median duration of hospitalization was 31 d (range 16–60). Conclusions: After a median follow‐up of 12 months from ASCT, nine patients are alive in continuous CR and 13 died from AML relapse. Median survival from diagnosis and disease‐free survival (DFS) was 19 and 14 months, respectively. Our data show that ASCT with a standard conditioning regimen is feasible in AML patients aged more than 60 yr. Toxicity and hemopoietic recovery do not substantially differ from those observed in young adults. DFS and overall survival (OS) duration are encouraging, but a longer follow up is needed on a larger series of patients.
Increase of haemoglobin levels by anti‐retroviral therapy is associated with a decrease in immune activation
Authors: Sarcletti Mario , Quirchmair Gisela , Weiss Günter , Fuchs Dietmar , Zangerle Robert
Published: March 2003

Abstract: Design: We evaluated whether an increase in haemoglobin levels in the first 6 months of effective anti‐retroviral therapy (ART) is associated with a decrease in immune activation. To reduce confounding factors only men (n = 35) and patients not receiving agents known to enhance haematopoiesis or patients without diseases that might suppress haematopoiesis were included. Simultaneously parameters of iron metabolism and cofactors for haematopoiesis were analysed. Results: A median baseline haemoglobin level of 139 g L−1 increased to 149 g L−1 at month 6 of ART (P < 0.001). At baseline low haemoglobin levels were strongly associated with high neopterin concentrations (r = − 0.64, P < 0.001), and much less correlated to high HIV‐1 RNA levels (r = − 0.41, P < 0.05) and to a lower CD4+ cell count (r = 0.33, P < 0.05). The change of neopterin levels during the study period correlated with the relative change in haemoglobin levels, r = − 0.35, P = 0.03, whereas no such correlations were found for the change of HIV‐1 RNA levels and the CD4 cell count. A logistic regression analysis revealed that the change of neopterin and soluble transferrin receptors concentrations are independently associated with an increase of haemoglobin levels of more than 15 g L−1. Conclusion: Our study supports a cause–effect relationship between immune activation and anaemia in HIV‐infected patients. Treatment of patients with ART decreases virus load, which may thereby result in silencing of immune effector activity thus ameliorating anaemia by reversing the anti‐proliferative effects of cytokines towards erythroid progenitors and the iron withdrawal strategy of the immune system.
Langerhans cell migration is modulated by N‐sulfated glucosamine moieties in heparin
Authors: O'Sullivan G. M. , Boswell C. M. , Halliday G. M.
Published: March 2003

Abstract: Dendritic cell (DC) migration into and out of tissues is important for the generation of primary immune responses to antigens encountered in tissues. In order to study the mechanisms involved in DC migration we used a skin explant system and quantitated the number of Langerhans cells (LC), which are immature precursors of DC in skin‐draining lymph nodes, remaining in the epidermis in response to incubation with various biomolecules. This paper shows that LC trafficking in epidermis is a metabolically active process that is modulated by heparin, specifically by N‐sulfated glucosamine moieties in heparin. This is the first demonstration of structural specificity in the biochemical requirements for DC migration in a tissue and therefore is important to understanding DC migration in general.
Spontaneous keratinocyte cell lines representing early and advanced stages of malignant transformation of the epidermis
Authors: Proby C. M. , Purdie K. J. , Sexton C. J. , Purkis P. , Navsaria H. A. , Stables J. N. , Leigh I. M.
Published: March 2003

Abstract: A unique series of epidermal cell lines representing different stages of malignant transformation were spontaneously derived from a single adult immunosuppressed individual. Four keratinocyte lines (PM1–4) established from forehead skin are here compared with 4 squamous cell carcinoma (SCC) lines (MET1–4) derived respectively from a primary cutaneous tumour, two local recurrences and a distant metastasis of invasive SCC. Despite altered growth properties, the PM lines retained many features of normal keratinocytes including keratin phenotype, differentiation capacity and non‐tumorigenicity in athymic mice. In contrast, from early passage, the MET lines displayed markedly reduced growth requirements, abnormal differentiation, aberrant K18 expression and tumorigenicity in athymic mice. The abnormal keratin profile of individual MET lines closely reflected the keratin phenotype of the tumour of origin. Although unusual HPV types were identified in the original tissue, there was no evidence of persistent virus within any cell line and it appears that HPV is not critical for maintenance of the immortal phenotype. The PM lines were distinctly different from invasive SCC lines and are likely to be useful for studies of mutations important early in neoplastic progression. The SCC series represent primary, recurrent and metastatic carcinoma. Availability of such a series from the same individual will facilitate genetic analysis of the malignant process.
Inhibition of keratinocyte growth in cell culture and whole skin culture by mast cell mediators
Authors: Huttunen M. , Hyttinen M. , Nilsson G. , Butterfield J. H. , Horsmanheimo M. , Harvima I. T.
Published: March 2003

Abstract: Mast cells are suggested to participate in regenerative processes, but their influence on epithelialization and wound healing has not been well studied. Since mast cells can be found in contact with epidermis in chronic inflammatory skin diseases and venous ulcers, the effect of mast cells on keratinocyte growth was studied. Keratinocytes were cultured in serum‐free conditions with (complete medium) or without (basal medium) epidermal growth factor (EGF) and bovine pituitary extract (BPE) to reach subconfluence in a 24‐well plate, and the cells were treated with different mast cell mediators histamine, heparin and tryptase, or lysate from HMC‐1 cells, a human leukemic mast cell line. Whole skin cultures were used as a model for in vitro wounds to study the effect of mast cells on epithelial outgrowth from skin specimens. Histamine inhibited 3H‐thymidine incorporation of keratinocytes dose‐dependently by 29% at 1 mM, and 89% at 5 mM histamine. In whole skin culture, histamine inhibited epithelial outgrowth dose‐dependently by 64% already at 0.1 mM histamine and maximally (91%) at 1 mM histamine. Heparin inhibited 3H‐thymidine incorporation dose‐dependently by up to 33% at 2 μg/ml in the absence, but not in the presence, of EGF/BPE. In contrast, in whole skin culture, heparin first inhibited the epithelial outgrowth by up to 27% at 2 μg/ml, but then reversed the inhibition to 30% stimulation at 200 μg/ml. Skin tryptase (0.0285 to 2.85 μg/ml) with or without heparin (0.5 to 20 μg/ml) did not affect thymidine incorporation in keratinocytes. Lysate from HMC‐1 cells, but not that from control, neuroblastoma cells, inhibited 3H‐thymidine incorporation in keratinocytes dose‐dependently, and maximal (47%) inhibition was reached with 16,700 lysed HMC‐1 cells/ml. In whole skin culture, HMC‐1 lysate inhibited the epithelial outgrowth by up to 36% at 67,000 lysed cells/ml. The results show that mast cells and their mediators are inhibitory to keratinocyte 3H‐thymidine incorporation and epithelial outgrowth in vitro, although, the inhibitory effect of histamine was seen at high concentrations suggesting a requirement for close morphologic vicinity of mast cells to keratinocytes. Thus, mast cells are assumed to control epidermal regeneration and to impair epithelialization of chronic ulcers.
Molecular genetics of pseudoxanthoma elasticum
Authors: Ringpfeil F. , Pulkkinen L. , Uitto J.
Published: March 2003

Abstract: Pseudoxanthoma elasticum (PXE), a systemic heritable connective tissue disorder, is characterized by progressive calcification of elastic structures in the skin, the eyes and the cardiovascular system, with considerable intra‐ and interfamilial phenotypic variability. Recently, underlying genetic defects have been identified in the ABCC6 gene, which resides on the chromosomal locus 16p13.1 and encodes the MRP6 protein, a member of the ATP‐binding cassette (ABC) family of proteins. The affected individuals are homozygous or compound heterozygous for a spectrum of genetic lesions, including nonsense and missense mutations, or deletions and splice‐site alterations, confirming the autosomal recessive nature of this condition. Analysis of the deduced primary sequence suggests that MRP6 is a transmembrane transporter, but its function has not been delineated yet. Surprisingly, however, MRP6 is expressed primarily, if not exclusively, in the liver and the kidneys, suggesting that PXE may be a primary metabolic disorder with secondary involvement of elastic fibers. Identification of mutations in the ABCC6 gene in PXE provides a means for prenatal and presymptomatic testing in families at risk for recurrence. DNA‐based analyses will also identify heterozygous carriers who may be at risk for development of limited manifestations of the disease as a result of compounding genetic factors and/or environmental modifiers.
Shortened telomere length is demonstrated in T‐cell subsets together with a pronounced increased telomerase activity in CD4 positive T cells from blood of patients with mycosis fungoides and parapsoriasis
Authors: Wu K.‐D. , Hansen E. R.
Published: March 2003

Abstract: We have recently demonstrated that telomerase activity is increased and telomere length shortened in lymphocytes from peripheral blood of patients with cutaneous T‐cell lymphoma. In order to determine which cell type has increased telomerase activity and shortened telomere length, CD4+, CD8+, CLA+ CD3+ and CLA− CD3+ T cells were isolated from peripheral blood of 25 patients, including 15 patients with mycosis fungoides and 10 patients with parapsoriasis. Eleven healthy individuals were used as controls; CD19+ B cells were separated from each individual as an internal control. The results showed that the increased telomerase activity was significantly predominating in the CD4+ T‐cell subset. Significantly shortened telomere length was found in CD4+ and CD8+ T‐cell subsets from the patients compared with the same cell subsets obtained from healthy individuals. However, no difference was observed between the subsets; CD19+ B cells collected from patients and healthy control individuals had similar telomerase activity and telomere length which was significantly different from the values found in T cells. The telomere length was significantly shorter in CLA+ CD3+ subset than in CLA− CD3+ subset. Interestingly, increased telomerase activity and shortened telomere length was also detected in CD4+ T cells from patients with parapsoriasis indicating that alteration of telomerase activity and telomere length in CD4+ T cells is an early event in the pathogenesis of cutaneous T‐cell lymphoma. Thus, the results indicate that a significant high level of telomerase activity and shortened telomere length frequently occur in T cells of patients with CTCL and may reflect tumorigenesis.
Basis for the permeability barrier abnormality in lamellar ichthyosis
Authors: Elias Peter M. , Schmuth Matthias , Uchida Yoshikazu , Rice Robert H. , Behne Martin , Crumrine Debra , Feingold Kenneth R. , Holleran Walter M. , Pharm D.
Published: March 2003

Abstract: The basis for the permeability barrier abnormality in lamellar ichthyosis (LI) is not known. LI is caused by mutations in the gene that encodes the enzyme, transglutaminase 1 (TGI), which is responsible for assembly of the cornified envelope (CE). TG1 also has been suggested recently to catalyze the covalent attachment of omega‐hydroxyceramides (omega‐OHCer) to the CE, forming the corneocyte‐lipid envelope (CLE). We first assessed the barrier function and the permeability pathway of the water‐soluble tracer, colloidal lanthanum, across the stratum corneum (SC) in patients with LI with absent (n = 4) or low (n = 2) TG1 activity/protein. Increased movement of tracer through the SC correlated with increased transcutaneous water loss, and tracer remained restricted to the SC interstices. Enhanced extracellular permeability, in turn, was explicable by truncation and fragmentation of extracellular lamellar membrane arrays. The resultant clefts in the SC interstices represent the likely pathway for increased water permeability. Moreover, tracer movement remained restricted to the interstices, despite the demonstration of increased corneocyte fragility associated with widespread variations in CE structure. Regardless of variability in CE structure, however, CLE structure and bound omega‐OHCer content were normal. The normal CLE in LI may explain both the restriction of tracer to the SC interstices, as well as the presence of foreshortened membrane arrays with near‐normal interlamellar dimensions. Finally, the demonstration of a normal CLE in LI also raises questions about the putative role of TG1 in forming the CLE. These results demonstrate: (1) the extracellular nature of increased permeability in LI; (2) discontinuities in extracellular membrane structures that account for the enhanced permeability in LI; (3) that these membrane abnormalities are both associated with and explained by abnormalities in the subjacent CE scaffold; and (4) an intact CLE is present in LI, despite abnormalities in the CE, which may restrict water movement to the SC interstices in LI.
Nitric oxide‐peroxynitrite‐poly(ADP‐ribose) polymerase pathway in the skin
Authors: Virág László , Szabó Éva , Bakondi Edina , Bai Péter , Gergely Pál , Hunyadi János , Szabó Csaba
Published: March 2003

Abstract: In the last decade it has become well established that in the skin, nitric oxide (NO), a diffusable gas, mediates various physiologic functions ranging from the regulation of cutaneous blood flow to melanogenesis. If produced in excess, NO combines with superoxide anion to form peroxynitrite (ONOO–), a cytotoxic oxidant that has been made responsible for tissue injury during shock, inflammation and ischemia‐reperfusion. The opposite effects of NO and ONOO– on various cellular processes may explain the ‘double‐edged sword’ nature of NO depending on whether or not cellular conditions favour peroxynitrite formation. Peroxynitrite has been shown to activate the nuclear nick sensor enzyme, poly(ADP‐ribose) polymerase (PARP). Overactivation of PARP depletes the cellular stores of NAD+, the substrate of PARP, and the ensuing ‘cellular energetic catastrophy’ results in necrotic cell death. Whereas the role of NO in numerous skin diseases including wound healing, burn injury, psoriasis, irritant and allergic contact dermatitis, ultraviolet (UV) light‐induced sunburn erythema and the control of skin infections has been extensively documented, the intracutaneous role of peroxynitrite and PARP has not been fully explored. We have recently demonstrated peroxynitrite production, DNA breakage and PARP activation in a murine model of contact hypersensitivity, and propose that the peroxynitrite‐PARP route represents a common pathway in the pathomechanism of inflammatory skin diseases. Here we briefly review the role of NO in skin pathology and focus on the possible roles played by peroxynitrite and PARP in various skin diseases.
Production of acute and chronic itch with histamine and contact sensitizers in the mouse and guinea pig
Authors: Laidlaw Anita , Flecknell Paul , Rees Jonathan L.
Published: March 2003

Abstract: Itch is a major symptom of skin disease and is poorly understood, in part due to the lack of adequate small animal models. We show, using iontophoresis of histamine and capsaicin, that it is possible to induce scratching behaviour in both guinea pig and mouse. Use of iontophoresis may obviate the problems of induction of pain as well as itch when injection is used. The behavioural response to capsaicin, however, differs from that seen with histamine, raising the possibility that the use of scratch counts as a method of measuring itch severity needs to be set in the context of other responses. Naloxone partly inhibits scratching in mouse and guinea pig due to histamine. We also show that contact sensitization with 2‐4 dinitrochlorobenzene (DNCB) can be used as a simple assay for chronic itch allowing study of scratching over at least a 15‐h period. The characteristics of scratching (but not the time course) induced with DNCB are similar to those seen with histamine.
Laminin 10/11: an alternative adhesive ligand for epidermal keratinocytes with a functional role in promoting proliferation and migration
Authors: Pouliot Normand , Saunders Nicholas A. , Kaur Pritinder
Published: March 2003

Abstract: We have investigated the expression and function of the isoforms of laminin bearing the α5 chain, i.e. laminin‐10/11 in neonatal and adult human skin. By immunostaining human skin derived from a variety of anatomic sites, we found that the laminin‐α5 chain is expressed abundantly in the basement membrane underlying the interfollicular epidermis and the blood vessels in the dermis. Interestingly, while the expression level of the well‐studied laminin‐5 isoform did not change significantly with age, laminin‐10/11 (α5 chain) appeared to decrease in the basement membrane underlying the epidermis, in adult skin. In contrast, the levels of laminin‐10/11 in the basement membrane underlying blood vessels remained unchanged in neonatal vs. adult skin. Importantly, in vitro cell adhesion assays demonstrated that laminin‐10/11 is a potent adhesive substrate for both neonatal and adult keratinocytes and that this adhesion is mediated by the α3β1 and α6β4 integrins. Adhesion assays performed with fractionated basal keratinocytes showed that stem cells, transit amplifying cells and early differentiating cells all adhere to purified laminin‐10/11 via these receptors. Further, laminin‐10/11 provided a proliferative signal for neonatal foreskin keratinocytes, adult breast skin keratinocytes, and even a human papillomavirus type‐18 transformed tumorigenic keratinocyte cell line in vitro. Finally, laminin‐10/11 was shown to stimulate keratinocyte migration in an in vitro wound healing assay. These results provide strong evidence for a functional role for laminin‐10/11 in epidermal proliferation during homeostasis, wound healing and neoplasia.
Energy reserves and reproductive investment in male three‐spined sticklebacks, Gasterosteus aculeatus
Authors: Huntingford F. A. , Chellappa S. , Taylor A. C. , Strang R. H. C.
Published: March 2003

Abstract – In a 6‐month study of male sticklebacks from a single cohort of an annual population, energy reserves (carbohydrate, lipid and protein) in the carcass and gonad were found to increase from December to May. Energy reserves in the liver increased from December to March before declining between March and April, a period of marked gonadal growth in this population. Examination at the individual level of the interrelationships among the various components of the energy store (corrected for differences in body size) identified overall body condition as a major component of variation and highlighted a negative relationship between energetic investment in gonadal tissue and energy reserves in various somatic compartments. Early in the season, relative gonadal weight was largely unrelated to somatic energy reserves, but from March onwards there was a negative correlation between these two variables. These data suggest that a significant flux of glycogen and lipid from the somatic to the gonadal tissue occurs in male sticklebacks in the period just prior to the breeding season, with the extent of gonadal growth varying between individuals and occurring at the expense of overall nutritional status.Note
Production and predation rates in a cannibalistic Arctic char (Salvelinus alpinus L.) population
Authors: Finstad A. G. , Jansen P. A. , Langeland L.
Published: March 2003

Abstract – Production of cannibalistic Arctic char (Salvelinus alpinus (L.)) and their prey conspecifics were estimated in an allopatric population, situated in an alpine ultraoligotrophic lake in central Norway. Estimates of population abundance were obtained by mark recapture and successive removal experiments. Assuming stable age distribution, we estimated mean yearly biomass (±95% CL) of char in sampled size groups vulnerable to predation (60 to 150 mm) to 7.93 (5.11–14.30) kg · ha−1. Similarly, mean yearly biomass (±95% CL) of cannibalistic char (> 250 mm) was estimated to 0.62 (0.50–1.06) kg · ha−1. Annual production (±95% CL) of char in length groups 60 to 150 mm was estimated to be 4.31 (2.74–8.03) kg · ha−1 · year−1, and production (±95% CL) of cannibalistic char to 0.19 (0.15–0.33) kg · ha−1 · year−1. Depending on the food conversion factor (set to vary from 0.1 to 0.4), the cannibalistic char removed from 10% to 40% of the production of char in sampled size‐classes vulnerable to predation yearly. The overall ecological efficiency in energy transfer between the prey and predator population was 4.4%.
Seasonal and spatial variation in a prairie stream‐fish assemblage
Authors: Ostrand K. G. , Wilde G. R.
Published: March 2003

Abstract – Stream‐fish assemblage and environmental data for 13 sites in the upper Brazos River, Texas, USA during 1997 and 1998 were used to assess the relationship between environmental conditions, and seasonal and spatial variation in fish species abundance and distribution patterns. There was considerable spatial variation in environmental conditions among sites. Spatial variation in species diversity and species composition was related to variation in conductance (salinity), depth and current velocity among sites and streams. Species diversity increased downstream and species composition shifted from primarily cyprinodontids upstream to cyprinids downstream. Among all dominant species, spatial components of variation in fish abundance were greater than seasonal components, suggesting that assemblage structure is determined more by average or persistent differences in environmental conditions among sites than by seasonal variation in environmental conditions.
Current conservation genetics: building an ecological approach to the synthesis of molecular and quantitative genetic methods
Authors: Moran P.
Published: March 2003

AbstractAlthough neutral molecular markers have long been important tools for describing genetic variation in threatened fish species, many of the most critical questions in conservation relate more to quantitative genetic variation than to neutral markers. Quantitative genetic studies are typically expensive and time‐consuming to conduct, especially in some of the long‐lived vertebrates of conservation concern. The present review of recent literature in fish conservation genetics examines the traditional role of molecular studies in describing conservation units and providing indirect inference about local adaptation and adaptive potential. Of special interest are approaches that use a combination of molecular and quantitative genetic methods. Such studies are likely to provide important new insights into many conservation‐related problems. The review also explores how increasing interest in non‐neutral molecular markers is contributing to our understanding of the geographic scale and evolutionary importance of local adaptation in threatened populations. It is increasingly clear that advanced genetic technologies for the exploration of neutral and non‐neutral molecular variation are leading to a fundamental shift in the way complex phenotypic traits are studied. This new synthesis of methods will have dramatic implications for fish conservation genetics and biology in general.
Diel microhabitat use of age‐0 bull trout in Indian Creek, Washington
Authors: Polacek M. C. , James P. W.
Published: March 2003

Abstract  – We studied diel microhabitat use at the focal point of age‐0 bull trout, Salvelinus confluentus, in Indian Creek, Washington during mid‐summer and fall of 1997. Microhabitat variables included water depth and velocity, distance from the stream bottom, habitat and refuge use, substrate type, and substrate embeddedness. Age‐0 fish were located over fines and gravel substrates in shallow, low‐velocity water near stream margins, but were located in shallower water at night. Bull trout were highly associated with loose substrate, and used the substrate interstices for refuge cover. Diurnal bull trout counts decreased and no age‐0 fish were observed after 15 September at water temperatures below 6.1 °C. Nocturnal counts remained relatively constant throughout the study. Our results suggest that age‐0 bull trout surveys be conducted at night when summer water temperatures begin to decline.
Oral status of 35 subjects with eating disorders – A 1‐year study
Authors: Öhrn Rolf , Angmar‐Månsson Birgit
Published: March 2003

The aim was to record changes over time in the oral status of subjects with diagnosed eating disorders. The outpatient psychiatrist had referred to the hospital dental clinic 35 women (19–47 yr, median 27 yr) with eating disorders, diagnosed according to DSM III‐R criteria. At the baseline examination, dental, medical and dietary histories were taken, and intra‐oral clinical and radiographic examinations were supplemented by intra‐oral photographs, study casts and salivary analysis. The subjects were re‐examined 1 yr later. Together, the investigators assessed progression of tooth wear blindly by comparing coded study casts from the baseline and 12‐month examinations. Progression of erosive tooth wear was recorded in almost half of the subjects. Several subjects had low unstimulated salivary flow rates (<0.1 ml/min) and very high counts of mutans streptococci and lactobacilli, both at baseline and 1 yr later. The flow rates for paraffin‐stimulated saliva at baseline were significantly lower for subjects with progression of erosive tooth wear than for those without. Because of the increased susceptibility to both caries and erosion, patients with eating disorders should be encouraged to have regular dental check‐ups. Test of salivary flow may serve as an indicator of patients' risk of progression of erosive tooth wear.
Adhesive bonding of titanium with a thione‐phosphate dual functional primer and self‐curing luting agents
Authors: Taira Yohsuke , Yanagida Hiroaki , Matsumura Hideo , Yoshida Keiichi , Atsuta Mitsuru , Suzuki Shiro
Published: March 2003

A consistent bond between the metal framework and the luting agent is desired when resin‐bonded prostheses are constructed with titanium. The purpose of this study was to evaluate three different metal primers on titanium bonding. Two sources of titanium (machined 99.9% titanium and cast Titan Ingot JS2) were used. Disk specimens were bonded with eight combinations of three primers and two luting agents (Panavia 21 and Super‐Bond C&B), including two controls. Shear bond strengths were determined after 24‐h water storage and after 10,000 cycles of thermocycling. Bond strengths were influenced by thermocycling, primer, luting agent and their combinations, but no significant differences were found between the machined 99.9% titanium and the cast ingot. The thione‐phosphate dual functional primer (Alloy Primer) was comparable to the phosphate primer (Cesead II Opaque Primer) and the thiophosphate primer (Metal Primer II) for bonding the titanium metals examined. The most durable bond was obtained in three combinations of these primers and one luting agent (Super‐Bond C&B).
Age‐period‐cohort analysis of dentist use in Spain from 1987 to 1997. An analysis based on the Spanish National Health Interview Surveys
Authors: Bravo Manuel
Published: March 2003

The aim was to determine whether the effects of age, period (from 1987 to 1997), or cohort (birth‐cohort) are the most plausible explanations for the variation in dentist utilisation over time in Spain. On the basis of three sequential Spanish National Health Surveys, the trend in the percentage of people visiting the dentist within the previous 3 months was analysed by means of a standard cohort table and by the Clayton & Schifflers method. The percent of people visiting the dentist grew from 13.6% in 1987, to 17.2% in 1997. Visual examination of the cohort table, together with the fit of the age‐cohort model, indicates that cohort succession may be the best explanation for the increase. The younger the cohort, the higher the dental visit rate; this relationship was strongest for people born after 1977. The model also indicates that the rates of dental visiting tend to increase up to middle age, and then tend to decrease with age.
The extracellular matrix of muscle – implications for manipulation of the craniofacial musculature
Authors: Lewis M. P. , Machell J. R. A. , Hunt N. P. , Sinanan A. C. M. , Tippett H. L.
Published: March 2003

Successful adaptation of craniofacial skeletal muscle is dependent upon the connective tissue component of the muscle. This is exemplified by procedures such as distraction histo/osteogenesis. The mechanisms underlying remodelling of intramuscular connective tissue are complex and multifactorial and involve extracellular matrix (ECM) molecules, receptors for the ECM (integrins) and enzymes that remodel the ECM (MMPs). This review discusses the current state of knowledge and clinical implications of connective tissue biology as applied to craniofacial skeletal muscle.
Dipeptidyl‐peptidase II and cathepsin B activities in amelogenesis of the rat incisor
Authors: Smid Joseph Ralph , Young William George , Monsour Paul Andrew
Published: March 2003

A body of published evidence suggests that a significant portion of enamel matrix protein synthesized by ameloblasts localises in the lysosomal‐endosomal organelles of these enamel organ cells. Little is known regarding the lysosomal proteolytic activities during amelogenesis. The aims of this study were to detect and measure the activities of lysosomal peptidases cathepsin B (E.C. 3.4.22.1) and dipeptidyl‐peptidase II (E.C. 3.4.14.2) in the enamel organ of the rat incisor and to ascertain whether rat enamel matrix proteins are degraded by these peptidases in vitro. Whole enamel organs were dissected from rat mandibular incisors. Enamel protein was also collected from the rat teeth. Analysis indicated that the rat incisor enamel organs contained specific activities of both dipeptidyl‐peptidase II and cathepsin B at levels comparable with those of kidney which is rich in both these lysosomal peptidases. Gel electrophoresis and immunoblotting demonstrated that both cathepsin B and dipeptidyl‐peptidase II were able to substantially degrade the rat enamel proteins in vitro. Based on these observations, we propose that lysosomal proteases have roles in amelogenesis in the intracellular degradation of amelogenins.
Periodontal tissue regeneration by combined applications of recombinant human osteogenic protein‐1 and bone morphogenetic protein‐2. A pilot study in Chacma baboons (Papio ursinus)
Authors: Ripamonti Ugo , Crooks Jean , Petit Jean‐Claude , Rueger David C.
Published: March 2003

Native and recombinant human bone morphogenetic/osteogenic proteins (BMPs/OPs) singly initiate bone induction in vivo. The finding of synchronous but spatially different BMPs/OPs expression during periodontal tissue morphogenesis suggests novel therapeutic approaches using morphogen combinations based on recapitulation of embryonic development. Twelve furcation defects prepared in the first and second mandibular molars of three adult baboons (Papio ursinus) were used to assess whether qualitative histological aspects of periodontal tissue regeneration could be enhanced and tissue morphogenesis modified by combined or single applications of recombinant hOP‐1 and hBMP‐2. Doses of BMPs/OPs were 100 µg of each protein per 1 g of insoluble collagenous bone matrix as carrier. Approximately 200 mg of carrier matrix was used per furcation defect. Undecalcified sections cut for histological analysis 60 d after healing of hOP‐1‐treated specimens showed substantial cementogenesis with scattered remnants of the collagenous carrier. hBMP‐2 applied alone induced greater amounts of mineralized bone and osteoid when compared to hOP‐1 alone or to combined morphogen applications. Combined applications of hOP‐1 and hBMP‐2 did not enhance alveolar bone regeneration or new attachment formation over and above the single applications of the morphogens. The results of this study, which is the first to attempt to address the structure‐activity relationship amongst BMP/OP family members, indicate that tissue morphogenesis induced by hOP‐1 and hBMP‐2 is qualitatively different when the morphogens are applied singly, with hOP‐1 inducing substantial cementogenesis. hBMP‐2 treated defects, on the other hand, showed limited cementum formation but a temporal enhancement of alveolar bone regeneration and remodelling. The demonstration of therapeutic mosaicism in periodontal regeneration will require extensive testing of ratios and doses of recombinant morphogen combinations for optimal tissue engineering in clinical contexts.
The heat shock response of Fusobacterium nucleatum
Authors: Skår Cecilie Kristin , Bakken Vidar
Published: March 2003

The heat‐shock response of the oral Gram‐negative bacterium Fusobacterium nucleatum was examined. Different strains of F. nucleatum were grown at 37°C, 42°C and 48°C in the presence of [35S]methionine. Cellular proteins synthesised after shifts to higher temperatures were analysed by SDS‐PAGE and autoradiography. Strains ATCC 10953, F1, F3 and Fev1 exhibited heat‐shock response, and major proteins were observed at 60, 70 and 90 kDa, but increased protein synthesis was also observed for other proteins. Immunoblot analysis, using a panel of antibodies directed to epitopes on different known heat‐shock proteins revealed cross‐reactive proteins, indicating homology between Escherichia coli, Mycobacterium leprae and F. nucleatum heat shock proteins.
BMP‐7 gene transfer to inflamed ferret dental pulps
Authors: Rutherford R. Bruce
Published: March 2003

In vivo and ex vivo gene transfer are being developed for localized skeletal regeneration. These strategies for tissue regeneration were tested in an adult ferret model of vital pulp therapy. In this model a reversible pulpitis was induced first. Then after 3 d, the pulps were directly infected with recombinant virus or implanted with ex vivo transduced autologous dermal fibroblasts. The genome of the recombinant adenovirus contained a full‐length cDNA encoding mouse bone morphogenetic protein (BMP)‐7 (AdBMP7) or bacterial β‐galactosidase cDNA (AdlacZ). The BMP‐7, but not lacZ, ex vivo transduced dermal fibroblasts induced reparative dentinogenesis with apparent regeneration of the dentin‐pulp complex. In vivo infection with AdBMP‐7 failed to produce reparative dentin in all cases. Ex vivo gene transfer of BMP‐7 may be an effective method for inducing dentin regeneration in teeth with reversible pulpitis.