bluuebunny/crossref_metadata_2025 huggingface dataset:
Re-investigation of the structure and crystal chemistry of the Bi2O3–W2O6 `type (Ib)' solid solution using single-crystal neutron and synchrotron X-ray diffraction
Authors: Sharma Neeraj , Macquart Rene B. , Avdeev Maxim , Christensen Mogens , McIntyre Garry J. , Chen Yu-Sheng , Ling Chris D.
Published: February 2010

Single crystals of composition Bi35.66W4.34O66.51 (or Bi8.2WO15.3, bismuth tungsten oxide), within the type (Ib) solid-solution region of the Bi2O3–WO3 system, were synthesized using the floating-zone furnace method. Synchrotron X-ray and neutron single-crystal diffraction data were used to confirm the previously tentative assignment of the room-temperature space group as I41. Fourier analysis of the combined X-ray and neutron datasets was used to elucidate and refine fully the cation and anion arrays for the first time. The mixed cation site M1 is shown to be coordinated by eight O atoms in an irregular cube when M = Bi, and by six O atoms in an octahedron when M = W. The resulting disorder in the average structure around M1 is discussed in the context of experimentally observed oxide-ion conductivity.
Proton-transfer dynamics in the (HCO3−)2dimer of KHCO3from Car–Parrinello and path-integrals molecular dynamics calculations
Authors: Dopieralski Przemyslaw D. , Latajka Zdzislaw , Olovsson Ivar
Published: February 2010

The proton motion in the (HCO_3^-)2dimer of KHCO3at 298 K has been studied with Car–Parrinello molecular dynamics (CPMD) and path-integrals molecular dynamics (PIMD) simulations. According to earlier neutron diffraction studies at 298 K hydrogen is disordered and occupies two positions with an occupancy ratio of 0.804/0.196. A simulation with only one unit cell is not sufficient to reproduce the disorder of the protons found in the experiments. The CPMD results with four cells, 0.783/0.217, are in close agreement with experiment. The motion of the two protons along the O...O bridge is highly correlated inside one dimer, but strongly uncoupled between different dimers. The present results support a mechanism for the disorder which involves proton transfer from donor to acceptor and not orientational disordering of the entire dimer. The question of simultaneous or successive proton transfer in the two hydrogen bonds in the dimer remains unanswered. During the simulation situations with almost simultaneous proton transfer with a time gap of around 1 fs were observed, as well as successive processes where first one proton is transferred and then the second one with a time gap of around 20 fs. The calculated vibrational spectrum is in good agreement with the experimental IR spectrum, but a slightly different assignment of the bands is indicated by the present simulations.
Erythema multiforme, Stevens–Johnson syndrome and toxic epidermal necrolysis: Frozen‐section diagnosis
Authors: HOSAKA Hiroomi , OHTOSHI Shinpei , NAKADA Tokio , IIJIMA Masafumi
Published: February 2010

Stevens–Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) may be fatal. Although classified by body surface area skin detachment, initial stages of both may present with erythema multiforme (EM)‐like lesions. To diagnose and predict disease activity adequately as early as possible for patients revealing EM‐like lesions, we performed frozen‐section diagnosis. Thirty‐five patients clinically diagnosed as EM, SJS or TEN were biopsied to diagnose and predict disease progression within the initial‐visit day. Half of a histological section taken from a lesion was snap‐frozen and immediately cryostat‐sectioned, acetone‐fixed and stained with hematoxylin–eosin. Specimens were examined with light microscopy for presence of epidermal necrosis. A section from unaffected sites was also examined for 11 patients. Specimens were examined with light microscopy for presence of graft‐versus‐host reaction (GVHR)‐like findings: apoptotic keratinocytes and satellite cell necrosis. Epidermal necrosis was seen in nine patients. Initial diagnosis of the nine was one of overlap SJS‐TEN, four of SJS and four of EM, and final diagnosis of those was one of TEN, one of overlap SJS–TEN, four of SJS and three of EM. Dissociation between initial and final diagnosis was seen in three cases. GVHR‐like findings in the epidermis were observed in two patients finally diagnosed as overlap SJS–TEN and TEN. Frozen sections are useful not only to make a diagnosis of erythema multiforme but to assess a potential to exhibit more aggressive clinical behaviors (SJS or TEN).
Herpetiform pemphigus without anti‐desmoglein 1/3 autoantibodies
Authors: NAKASHIMA Hiroko , FUJIMOTO Manabu , WATANABE Rei , ISHIURA Nobuko , YAMAMOTO Akemi Ishida , HASHIMOTO Takashi , TAMAKI Kunihiko
Published: February 2010

We report a patient with herpetiform pemphigus who was negative for autoantibodies to desmoglein (Dsg)1 or 3. He had erythemas with vesicles lining the margins on the trunk and extremities. Histopathology revealed intraepidermal blister with prominent eosinophil infiltration. Direct and indirect immunofluorescence demonstrated the presence of depositing and circulating immunoglobulin (Ig)G autoantibodies, but no IgA antibodies, to keratinocyte cell surface. Nonetheless, neither anti‐Dsg1 nor Dsg3 antibodies were detected by enzyme‐linked immunosorbent assay. Immunoblotting using human epidermal extracts also showed no reactivity with known intraepidermal or epidermal–dermal junctional substances. Immunoelectronmicroscopy revealed the reactivity on the portion of keratinocyte cell surface but not on the desmosomes. This case suggests that non‐desmoglein antigen on keratinocyte cell surface can be targeted in some patients with this unusual variant of pemphigus.
What’s new in bullous pemphigoid
Authors: UJIIE Hideyuki , SHIBAKI Akihiko , NISHIE Wataru , SHIMIZU Hiroshi
Published: February 2010

Bullous pemphigoid (BP) is the most common autoimmune blistering disease. BP patients have autoantibodies against type XVII collagen (COL17, also called BP180 or BPAG2), a type II transmembrane protein that spans the lamina lucida and projects into the lamina densa of the epidermal basement membrane. The non‐collagenous 16A domain of COL17 is considered to contain pathogenic epitopes of BP. The transfer of immunoglobulin (Ig)G from BP patients fails to cause blisters on mouse skin probably due to differences between humans and mice in the amino acid sequence of NC16A pathogenic epitope of COL17. Passive transfer of rabbit IgG antibodies against the murine homolog of human COL17 NC16A triggers immune reactions to COL17 in mice, including complement activation, mast cell degranulation and neutrophilic infiltration, resulting in dermal–epidermal separation. Recent studies using COL17‐humanized mice that express human COL17 but lack murine COL17 were the first to demonstrate the pathogenicity of anti‐COL17 human BP IgG autoantibodies in vivo. These new findings provide a greater understanding of BP pathomechanisms and facilitate the development of novel specific and efficient therapeutic strategies for BP.
Bullous pemphigoid as a dermadrome associated with spindle cell carcinoma of the gallbladder
Authors: UMEKOJI Ayano , TSURUTA Daisuke , INOUE Toru , NISHIMORI Takeo , ISHII Masamitsu
Published: February 2010

Skin disorders that appear in association with internal malignancies are called dermadromes. Bullous pemphigoid (BP), which is a major autoimmune disease of the skin, is considered to be a dermadrome, although there have been conflicting reports. We report a case of BP that preceded the diagnosis of an internal malignancy. Although we could not detect any malignancies on chest, abdominal or pelvic computer tomography on the first hospital admission, intensive screening on the third admission revealed a gallbladder malignancy. Laparoscopic cholecystectomy was performed. Histopathology showed a spindle cell carcinoma of the gallbladder. To the best of our knowledge, this is the first report of a spindle cell carcinoma of the gallbladder in a patient with BP.
Immune dysregulation of pemphigus in humans and mice
Authors: YOKOYAMA Tomoaki , AMAGAI Masayuki
Published: February 2010

Pemphigus vulgaris is an autoimmune blistering disease of the skin and mucous membranes that is caused by immunoglobulin (Ig)G autoantibodies against the cadherin‐type adhesion molecule desmoglein (Dsg)3 expressed on stratified epithelial cells. Interaction between antigen‐specific T and B cells, which is selectively achieved through T‐cell receptor/major histocompatibility complex–peptide complex association and subsequently corroborated by co‐stimulatory molecules such as CD40/CD154, is required for production of pathological anti‐desmoglein 3 antibody. Some genetically and environmentally susceptible individuals harbor desmoglein reactive B and T cells, and anti‐desmoglein antibodies were detected in their serum. Analysis of the anti‐desmoglein antibody clones derived from pemphigus patients or pemphigus model mice revealed that pathogenic antibodies mostly react with conformational epitopes on mature form desmogleins, whereas non‐pathogenic ones tend to react with non‐conformational epitopes. Surprisingly, antibodies to the Dsg1 precursor pro‐protein are also cloned from individuals without pemphigus. These observations suggest that active suppression by regulatory cell subsets is dominant in these susceptible individuals. In fact, Dsg‐reactive T‐inducible regulatory type 1 (Tr1) cells are readily detected in healthy carriers of pemphigus‐related human leukocyte antigen haplotypes, but rarely in pemphigus patients. These Tr1 cells can be functionally converted to T‐helper 2‐like cells which secrete interleukin‐2 by inactivation of Foxp3 through antisense oligonucleotides. Thus, delicate balance between self‐reactive lymphocytes and regulatory T cells may be a key element in determining whether individuals produce pathogenic antibodies and develop pemphigus phenotypes or not.
Epidermolysis bullosa acquisita: What’s new?
Authors: ISHII Norito , HAMADA Takahiro , DAINICHI Teruki , KARASHIMA Tadashi , NAKAMA Takekuni , YASUMOTO Shinichiro , ZILLIKENS Detlef , HASHIMOTO Takashi
Published: February 2010

Type VII collagen is an adhesion molecule of the extracellular matrix in epithelial basement membranes, and the main constituent of anchoring fibrils at the dermal–epidermal junction (DEJ). Autoimmunity against this protein is causing the rare organ‐specific epidermolysis bullosa acquisita (EBA). EBA is a rare acquired, heterogeneous, chronic blistering disease of skin disease of skin and mucous membranes characterized by subepidermal blisters and tissue‐bound as well as circulating autoantibodies to the DEJ. EBA has several distinct clinical presentations with other subepidermal bullous diseases, such as mainly dystrophic epidermolysis bullosa or bullous pemphigoid. The circulating immunoglobulin G autoantibodies for EBA react with a 290‐kDa dermal protein, type VII collagen, as detected by immunoblot analysis using dermal extracts. The pathogenicity of these autoantibodies has been demonstrated by experimental animal models, in which anti‐type VII collagen antibodies injected into a mouse produced an EBA‐like blistering disease in the animal. EBA cases often require high doses of systemic corticosteroids and a variety of immunosuppressants. Although treatment for EBA is frequently difficult and unsatisfactory, some therapeutic success has been reported with colchicine, dapsone, infliximab and i.v. immunoglobulin. In this review, we will focus on recent progress in our understanding of the clinical manifestations, the etiopathogenesis as well as the management of EBA.
Overview of epidermolysis bullosa
Authors: SAWAMURA Daisuke , NAKANO Hajime , MATSUZAKI Yasushi
Published: February 2010

Epidermolysis bullosa (EB) is classified into major types – EB simplex (EBS), junctional EB (JEB) and dystrophic EB (DEB) – on the basis of the level of tissue separation within the cutaneous basement membrane zone. Recent advances in research on EB have led to the identification of 10 genes responsible for EB. The Japanese Ministry of Health, Labor and Welfare has designated JEB and DEB, but not EBS, as specified diseases. However, EBS has a lethal variant and should also be registered as a specified disease. In the Third Consensus Meeting on the Diagnosis and Classification of EB held in Vienna in 2007, it was recommended that Kindler syndrome should be classified as a subtype of EB. Corrective gene therapy is the most ideal therapy for EB, but much more research is required before it can be developed and used in clinical practice. Cell‐based therapies using fibroblasts and bone marrow cells have recently attracted considerable attention.
Combustion metamorphism in the Nabi Musa dome: new implications for a mud volcanic origin of the Mottled Zone, Dead Sea area
Authors: Sokol E. , Novikov I. , Zateeva S. , Vapnik Ye. , Shagam R. , Kozmenko O.
Published: February 2010

Numerous high‐temperature combustion metamorphic foci within brecciated mainly calcareous sedimentary rocks in the Dead Sea area (the so‐called ‘Mottled Zone’ complexes) have been interpreted as resulting from in situ oxidation and ignition of dispersed organic matter in bituminous chalks. Geological, chemical and petrological data for the Nabi Musa dome, one of 15 Mottled Zone complexes data presented in this paper, suggest an alternative interpretation relating the Mottled Zone complexes to Pliocene–Pleistocene mud volcanism and the associated methane combustion. The geochemistry and mineralogy of sedimentary, combustion metamorphic, localized hydrothermally, altered rocks, and ignition foci marked by ultrahigh temperature (up to 1500 °C) pseudowollastonite–rankinite–nagelschmidtite‐bearing paralava, indicate that Nabi Musa is a fossil mud volcano, comprising a large diatreme edifice with brecciated sedimentary rocks in its main feeder. The mud volcanic event mobilized sediments of underlying Cretaceous strata from depths of at least 0.8 km, and the eruption was driven by hydrocarbon gases (predominantly methane), with gas flaming causing local combustion metamorphism. Besides ultrahigh temperature combustion metamorphic processes, ejected sedimentary rocks were subsequently altered by low‐temperature hydrothermal fluids from various sources, which produced specific rock compositions with local enrichments in Mg, Na, Cl and B. Later, carbonation almost completely replaced the original smectite‐bearing parent mud and preserved the edifice from erosion. The proposed mud‐volcanic origin of the Mottled Zone complexes may have implications for gas prospecting in the Levantine basin.
Hormonal and metabolic effects of morning or evening dosing of the dipeptidyl peptidase IV inhibitor vildagliptin in patients with type 2 diabetes
Authors: He Yan‐Ling , Valencia Jessica , Zhang Yiming , Schwartz Sherwyn L. , Ligueros‐Saylan Monica , Foley James , Dole William P.
Published: February 2010

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• Vildagliptin is an orally active, potent inhibitor of dipeptidyl peptidase IV and was developed for the treatment of type 2 diabetes.

• In clinical trials, once or twice daily dosing with vildagliptin (up to 100 mg day−1) has been shown to reduce endogenous glucose production and fasting plasma glucose in patients with type 2 diabetes.

• The comparative efficacy of vildagliptin under a morning vs. evening dosing regimen has not previously been determined.

WHAT THIS STUDY ADDS

• Once daily dosing with vildagliptin 100 mg for 28 days improved glycaemic control in patients with type 2 diabetes independent of whether vildagliptin was administered in the morning or evening.

• Morning or evening dosing with vildagliptin had similar effects on 24 h glycaemic control and plasma concentrations of the hormones insulin, glucagon and glucagon‐like peptide 1.

AIM

This randomized, double‐blind, crossover study compared post‐prandial hormonal and metabolic effects of vildagliptin, (an oral, potent, selective inhibitor of dipeptidyl peptidase IV [DPP‐4]) administered morning or evening in patients with type 2 diabetes.

METHODS

Forty‐eight patients were randomized to once daily vildagliptin 100 mg administered before breakfast or before dinner for 28 days then crossed over to the other dosing regimen. Blood was sampled frequently after each dose at baseline (day −1) and on days 28 and 56 to assess pharmacodynamic parameters.

RESULTS

Vildagliptin inhibited DPP‐4 activity (>80% for 15.5 h post‐dose), and increased active glucagon‐like peptide‐1 compared with placebo. Both regimens reduced total glucose exposure compared with placebo (area under the 0–24 h effect–time curve [AUE(0,24 h)]: morning −467 mg dl−1 h, P= 0.014; evening −574 mg dl−1 h, P= 0.003) with no difference between regimens (P= 0.430). Reductions in daytime glucose exposure (AUE(0,10.5 h)) were similar between regimens. Reduction in night‐time exposure (AUE(10.5,24 h) was greater with evening than morning dosing (−336 vs.−218 mg dl−1 h, P= 0.192). Only evening dosing significantly reduced fasting plasma glucose (−13 mg dl−1, P= 0.032) compared with placebo. Insulin exposure was greater with evening dosing (evening 407 µU ml−1 h; morning 354 µU ml−1 h, P= 0.050).

CONCLUSIONS

Both morning and evening dosing of once daily vildagliptin 100 mg significantly reduced post‐prandial glucose in patients with type 2 diabetes; only evening dosing significantly decreased fasting plasma glucose. Although evening dosing with vildagliptin 100 mg tended to decrease night‐time glucose exposure more than morning dosing, both regimens were equally effective in reducing 24 h mean glucose exposure (AUE(0,24 h)) in patients with type 2 diabetes.
Systemic bioavailability of hydrofluoroalkane (HFA) formulations of fluticasone/salmeterol in healthy volunteers via pMDI alone and spacer
Authors: Clearie Karine L. , Williamson Peter A. , Vaidyanathan Sriram , Du Bois Jeannine , Nell Haylene , Lipworth Brian J.
Published: February 2010

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• Seretide (GlaxoSmithKline, UK) is a combination inhaler which contains both fluticasone (FP) and salmeterol (SM).

• A generic fluticasone/salmeterol (FP/SM) combination inhaler (Neolab, UK) has recently been developed.

• Determination of therapeutic equivalence is essential in the development of generic products.

WHAT THIS STUDY ADDS

• This paper determines that in vitro fine particle dose may not predict pharmacokinetic and systemic pharmacodynamic outcomes.

• This paper shows that the generic and reference products were equivalent in terms of FP, but not SM pharmacokinetics.

• No significant differences were found between generic and reference products in terms of either adrenal suppression or hypokalaemia.

AIM To compare a test version of HFA fluticasone/salmeterol (FP/SM) combination inhaler (Neolab, UK) with the reference product Seretide (GlaxoSmithKline, UK).

METHODS An in vitro Anderson cascade impactor was used to compare the fine particle dose (<4.7 µm). Two separate randomized cross‐over studies were performed to compare the systemic bioavailability of test vs. reference (T vs. R) formulations of FP/SM 250/25 µg pMDI in healthy volunteers. In study 1 blood pharmacokinetic analysis using oral charcoal block was performed over 24 h following a single dose of four puffs via pMDI alone. In study 2 systemic bioactivity was measured following single doses of four and eight puffs via a spacer device: serum potassium (K+) to reflect SM, and overnight urinary cortisol : creatinine (OUCC) for FP. An early pharmacokinetic profile was also assessed over 120 min.

RESULTS The in vitro fine particle dose was similar for test vs. reference pMDI alone and via spacer. The results of both studies were consistent: No significant differences between formulations were seen in terms of FP kinetics. Analysis of SM kinetics revealed superiority of the test product. No significant dose–response or difference in T : R ratio was noted for OUCC. Fall in K+ revealed a significant dose–response with a non‐significant T : R ratio.

CONCLUSIONS The in vitro fine particle dose may not predict pharmacokinetic and systemic pharmacodynamic outcomes. Single dosing studies with fluticasone/salmeterol 250/25 µg via pMDI or with spacer showed pharmacokinetic equivalence with FP, but not SM. No significant difference between formulations was seen with either adrenal suppression or hypokalaemia.
Potential impact of cytochrome P450 3A5 in human liver on drug interactions with triazoles
Authors: Yamazaki Hiroshi , Nakamoto Minako , Shimizu Makiko , Murayama Norie , Niwa Toshiro
Published: February 2010

Cytochrome P450 3A is the main enzyme subfamily involved in the metabolism of a variety of marketed medicines. It is generally believed that the substrate specificity of polymorphic P450 3A5 is similar to that of the predominant P450 3A4 isoform, although some differences in catalytic properties have been found. It has been hypothesized that individuals withCYP3A5*1(P450 3A5 expresser) might clear the HIV protease inhibitor saquinavir, administered by mouth, more rapidly than subjects lacking functionalCYP3A5alleles. Enhanced midazolam hydroxylation and cyclosporin metabolism occur in anin vitroP450 3A5 system and in liver microsomes expressing P450 3A5 in the presence of thalidomide. However, inhibition constants (Ki) of three triazole anti‐fungal drugs (itraconazole, fluconazole, and voriconazole) for liver microsomal P450 3A5 are higher than for liver microsomal P450 3A4. To predict drug interactionsin vivo, we estimated increases of areas under the curves (AUC) dependent on polymorphic P450 3A5 expression, using both 1 +[Inhibitor] / Ki(recommended in US FDA guidance), and 1 +[Inhibitor]unbound/Ki(as recommended by Japanese MHLW Notice). Voriconazole would be expected to cause approximately a three‐fold higher increase in AUC in subjects withCYP3A5*3/*3than in those withCYP3A5*1/*3,especially when estimated using the FDA guidance. We conclude that drug interactions between marketed drugs may differ substantially between individuals with genetically distinct P450 3A5 catalytic functions.
Evaluation of Ki‐67 and Bcl‐2 antigen expression in breast carcinomas of women treated with raloxifene
Authors: Lopes‐Costa P. V. , Dos Santos A. R. , Dos Santos L. G. , Da Silva B. B.
Published: February 2010

Objectives:  To evaluate the effect of raloxifene on Ki‐67 and Bcl‐2 antigen expression in operable, stage II, oestrogen‐receptor‐positive invasive ductal breast carcinomas.

Materials and methods:  Twenty post‐menopausal women who had taken 60 mg of raloxifene daily for 28 days prior to definitive surgery were enrolled in the investigation. Two tumour samples were obtained by incisional biopsy during the study, one at the time of confirmation of diagnosis of invasive ductal carcinoma and evaluation of oestrogen receptor status, and the other 29 days later, at the time of definitive surgery. Immunohistochemistry was performed on tumour samples, prior to and after raloxifene treatment, to evaluate Ki‐67 and Bcl‐2 expression. Friedman and McNemar tests were used for statistical analysis of the data, significance being established at 5%.

Results:  Mean percentage of Ki‐67‐stained nuclei was 24.86 ± 2.95 prior to raloxifene treatment and 13.33 ± 1.52 after treatment (P < 0.001). Prior to raloxifene treatment, only 9/20 cases (45%) were classified as Bcl‐2‐positive, whereas after treatment, 17/20 (85%) were classified as Bcl‐2‐positive (P < 0.013).

Conclusions:  Raloxifene treatment significantly reduced Ki‐67 antigen expression and increased Bcl‐2 expression in breast carcinomas of post‐menopausal women.
Microplate spectrophotometry for high‐throughput screening of cytotoxic molecules
Authors: Tomelleri C. , Dalla Pellegrina C. , Chignola R.
Published: February 2010

Objectives:  High‐throughput chemical and biochemical technologies are now exploited by modern pharmacology and toxicology to synthesize a multitude of new molecules with bioactive potential, or to isolate them from living matter. Testing molecules in cell systems on a large scale, however, is a rate‐limiting step in drug discovery or in toxicity assessment. In this study, we developed a low‐cost high‐throughput method for first‐level screening of cytotoxic molecules.

Materials and methods:  We used microplate spectrophotometry to measure growth kinetics of human tumour cells that grow in suspension (Molt3) or adherent to the plastic surface of culture wells (HeLa) in standard RPMI medium. Cells were treated with colchicin, idarubicin or paclitaxel under various treatment schedules. The effects were quantified and compared with those measured by optical microscopy using the trypan blue dye exclusion method to reveal dead cells.

Results:  Proliferation kinetics of tumour cells can be quantified by measuring variations in optical densities of cell samples at 410 and 560 nm wavelengths. For cells that grow in suspension, one single reading at 730 nm may be sufficient to reconstruct growth curves that parallel those obtained by direct cell counting. Effects of the cytotoxic treatments could also be quantified and results compared very favourably with those obtained using standard techniques.

Conclusions:  Microplate spectrophotometry is a robust and sensitive method to monitor growth of animal cell populations both in the absence and in the presence of cytotoxic drugs. This method implements existing technologies and can be fully automated.
3‐O‐Methylfunicone, a metabolite produced by Penicillium pinophilum, modulates ERK1/2 activity, affecting cell motility of human mesothelioma cells
Authors: Buommino E. , Paoletti I. , De Filippis A. , Nicoletti R. , Ciavatta M. L. , Menegozzo S. , Menegozzo M. , Tufano M. A.
Published: February 2010

Objectives:  3‐O‐methylfunicone (OMF), a secondary metabolite produced by Penicillium pinophilum, affects cell proliferation and motility in a variety of human solid tumours. The aim of this study was to demonstrate whether OMF has the ability to arrest cell division and motility, in a human mesothelioma cell line. Malignant mesothelioma is an aggressive cancer that does not respond to standard therapies the cells of which are considered to be highly resistant to apoptosis.

Material and methods:  Cell motility and invasion were measured using a modified Boyden chamber. Gene expression was examined by RT‐PCR, while ERK1/2 was investigated by Western blot analysis. All experiments were also performed on primary cultures of mesothelial cells.

Results:  The present study shows that OMF inhibited motility of the NCI mesothelioma cell line by modulating ERK signalling activity, and affected αVβ5 integrin and MMP‐2 expression, inducing marked downregulation at both mRNA and protein levels. Substantial downregulation of VEGF gene expression was also demonstrated. These effects were not observed in normal mesothelial cell cultures.

Conclusion:  OMF may have potential as a naturally derived anti‐tumour drug for treatment of mesothelioma.
Analysis of radiation‐induced changes to human melanoma cultures using a mathematical model
Authors: Basse B. , Joseph W. R. , Marshall E. S. , Baguley B. C.
Published: February 2010

Objectives:  Tumour cells respond to ionizing radiation by cycle arrest, cell death or repair and possible regrowth. We have developed a dynamic mathematical model of the cell cycle to incorporate transition probabilities for entry into DNA replication and mitosis. In this study, we used the model to analyse effects of radiation on cultures of five human melanoma cell lines.

Materials and methods:  Cell lines were irradiated (9 Gy) prior to further culture and harvesting at multiple points up to 96 h later. Cells were fixed, stained with propidium iodide and analysed for G1‐, S‐ and G2/M‐phase cells by flow cytometry. Data for all time points were fitted to a mathematical model. To provide unique solutions, cultures were grown in the presence and absence of the mitotic poison paclitaxel, added to prevent cell division.

Results:  The model demonstrated that irradiation at 9 Gy induced G2‐phase arrest in all lines for at least 96 h. Two cell lines with wild‐type p53 status additionally exhibited G1‐phase arrest with recovery over 15 h, as well as evidence of cell loss. Resumption of cycling of surviving cells, as indicated by increases in G1/S and G2/M‐phase transitions, was broadly comparable with results of clonogenic assays.

Conclusions:  The results, combined with existing data from clonogenic survival assays, support the hypothesis that a dominant effect of radiation in these melanoma lines is the induction of long‐term cell cycle arrest.
Recombinant human midkine stimulates proliferation of articular chondrocytes
Authors: Zhang Z. H. , Li H. X. , Qi Y. P. , Du L. J. , Zhu S. Y. , Wu M. Y. , Lu H. L. , Yu Y. , Han W.
Published: February 2010

Objectives:  Midkine, a heparin‐binding growth factor, promotes population growth, survival and migration of several cell types, but its effect on articular chondrocytes remains unknown. The aim of this study was to investigate its role on proliferation of articular chondrocytes in vitro and in vivo.

Materials and methods:  Bromodeoxyuridine incorporation and MTT assays were performed to examine the proliferative effect of recombinant human midkine (rhMK) on primary articular chondrocytes. Activation of extracellular signal‐regulated kinase (ERK) and phosphatidylinositol 3‐kinase (PI3K) was analysed using western blot analysis. Systemic and local delivery of rhMK into mice and rats was preformed to investigate the proliferative effect of rhMK in vivo, respectively. Histological evaluation, including measurement of articular cartilage thickness, cell density, matrix staining and immunostaining of proliferating cell nuclear antigen was carried out.

Results:  rhMK promoted proliferation of articular chondrocytes cultured in a monolayer, which was mediated by activation of ERK and PI3K. The proliferative role of rhMK was not coupled to dedifferentiation of culture‐expanded cells. Consistent with its action in vitro, rhMK stimulated proliferation of articular chondrocytes in vivo when it was administered subcutaneously and intra‐articularly in mice and rats, respectively.

Conclusion:  Our results demonstrate that rhMK stimulates proliferation of primary articular chondrocytes in vitro and in vivo. The results of this study warrant further examination of rhMK for treatment of animal models of articular cartilage defects.
Patch size effects are more important than genetic diversity for plant–herbivore interactions in Brassica crops
Authors: HAMBÄCK PETER A. , BJÖRKMAN MARIA , HOPKINS RICHARD J.
Published: February 2010

1. Considerable evidence suggests that the diversity within plant communities may strongly affect the strength of species interactions, but the majority of studies only considered interspecific diversity.

2. This paper examines the effect of intraspecific genetic diversity within Brassica fields on two Brassica specialists, cabbage root fly, and diamondback moth, and on a parasitoid attacking diamondback moths. Genetic diversity was manipulated both in a replacement and an additive design.

3. Both herbivore densities and parasitism rates were higher in smaller plots, with limited responses to increased within‐plot diversity. All species showed variable densities across genotypes, and preference hierarchies were species specific.

4. Responses to plot size in root flies scaled with the diameter‐to‐area ratio, suggesting that patch detectability affected local density, whereas responses by diamondback moths and parasitoids deviated from this ratio. These species differences could be traced to differences in the residence time within patches, where diamondback moths typically spend longer and more variable time periods in patches than root flies.

5. The lack of response to genetic diversity by both herbivores suggests that egg‐laying rates are affected by decisions on the plant and not by attraction from a distance, neither to the plant itself nor the patch. Patterns of differential attack may then be due to different acceptability for studied genotypes.

6. Future theories on insect responses to spatial heterogeneity should focus on species traits and how traits interact with information landscapes in the field.
A novel frame‐shift deletion causing analbuminaemia in an Italian paediatric patient
Authors: Dagnino M. , Caridi G. , Marsciani M. , Bettocchi I. , Tassinari D. , Bernardi F. , Chiodo F. , Campagnoli M. , Galliano M. , Minchiotti L.
Published: February 2010

Eur J Clin Invest 2010; 40 (3): 281–284

Background  Analbuminaemia (OMIM #103600) is a rare autosomal recessive disorder manifested by the absence or severe reduction of circulating serum albumin in homozygous or compound heterozygous subjects. The trait is caused by a variety of mutations within the albumin gene.

Design  We report here the clinical and molecular characterization of a new case of congenital analbuminaemia in a 4‐year‐old Italian girl diagnosed on the basis of the low level of circulating albumin (= 10·0 g L−1). The albumin gene was screened by single‐strand conformation polymorphism and heteroduplex analysis and the mutated region submitted to DNA sequencing.

Results  The proband was found to be homozygous, and both parents heterozygous, for a novel deletion in exon 8 (c.920delT). The subsequent frame‐shift should have given rise to a putative polypeptide chain of 304 amino acid residues, which we could not identify in the proband’s serum.

Conclusions  A novel analbuminaemia causing mutation was identified and characterized at the clinical level in a child. The molecular diagnosis of the trait is based on the rapid localization of the mutation within the albumin gene by single‐strand conformation polymorphism and heteroduplex analysis, followed by DNA sequencing of the mutated region.
Cardiac hormones eliminate some human squamous lung carcinomas in athymic mice
Authors: Lenz A. , Sun Y. , Eichelbaum E. J. , Skelton W. P. , Pi G. , Vesely D. L.
Published: February 2010

Eur J Clin Invest 2010; 40 (3): 242–249

Background  Four cardiac hormones synthesized by the same gene, i.e. atrial natriuretic peptide, vessel dilator, long acting natriuretic peptide and kaliuretic peptide, have anticancer effects in vitro.

Materials and methods  These cardiac hormones were infused subcutaneously for 28 days with weekly fresh hormones at 0·3 nM kg−1 body weight in athymic mice bearing human squamous cell carcinomas.

Results  Vessel dilator, atrial natriuretic peptide and kaliuretic peptide each eliminated one in six (17%) of the human squamous cell lung carcinomas. Long‐acting natriuretic peptide, although it did not eliminate any of the human squamous cell lung carcinomas did decrease the volume of one carcinoma to only 2% (P < 0·0001) of the untreated carcinomas. The squamous cell lung carcinomas that were not eliminated, with the exception of the one LANP‐treated tumour that decreased to only 2% of the volume of the untreated cancers, grew rapidly but their growth velocity compared to controls decreased by 76%, 40%, 38% and 25% in the vessel dilator, atrial natriuretic peptide, kaliuretic peptide and long‐acting natriuretic peptide groups respectively (P < 0·05).

Conclusions  Three of four cardiac hormones synthesized by the atrial natriuretic peptide gene can eliminate human squamous cell lung carcinomas in athymic mice when treated subcutaneously for 4 weeks. The 4th cardiac hormone, i.e. long‐acting natriuretic peptide, decreased the volume of one squamous cell lung carcinoma to 2% of that of untreated animals, suggesting that it, too, has beneficial effects on squamous cell lung cancers.
Functional diversity is positively associated with biomass for lake diatoms
Authors: VOGT RICHARD J. , BEISNER BEATRIX E. , PRAIRIE YVES T.
Published: February 2010

1. Recent work has emphasised the benefit of using functional measures when relating biodiversity to ecosystem functioning. In this study, we investigated the extent to which functional and taxonomic diversity might be related to summed biovolume in community assemblages of 212 species of diatoms collected from 65 temperate lakes in western and central Quebec, Canada.

2. We quantified functional diversity as both the total path‐length of a functional dendrogram (FD) and the variance in species traits (TV) for a given community. Selected traits included both size and responses to a set of environmental variables known to be influential for diatom communities.

3. Species richness, as well as both FD and TV, was positively associated with total diatom biovolume at the level of the entire diatom community, suggesting that diversity in response types (particularly to total phosphorus and pH) is important for diatom community production.

4. Although functional measures of diversity did not provide enhanced explanatory power over species richness, we argue that an exploration of functional traits potentially allows greater insight into the mechanisms underlying biodiversity–ecosystem functioning relations, indicating which traits might be most influential in driving community biomass production.
Regulation of nitrous oxide emission associated with benthic invertebrates
Authors: STIEF PETER , SCHRAMM ANDREAS
Published: February 2010

1. A number of freshwater invertebrate species emit N2O, a greenhouse gas that is produced in their gut by denitrifying bacteria (direct N2O emission). Additionally, benthic invertebrate species may contribute to N2O emission from sediments by stimulating denitrification because of their bioirrigation behaviour (indirect N2O emission).

2. Two benthic invertebrate species were studied to determine (i) the dependence of direct N2O emission on the preferred diet of the animals, (ii) the regulation of direct N2O emission by seasonally changing factors, such as body size, temperature and availability and (iii) the quantitative relationship between direct and indirect N2O emission.

3. Larvae of the mayfly Ephemera danica, which prefer a bacteria‐rich detritus diet, emitted N2O at rates of up to 90 pmol Ind.−1 h−1 under in situ conditions and 550 pmol Ind.−1 h−1 under laboratory conditions. In contrast, larvae of the alderfly Sialis lutaria, which prefer a bacteria‐poor carnivorous diet, emitted N2O at invariably low rates of 0–20 pmol Ind.−1 h−1. The N2O emission rate of E. danica larvae was positively correlated with seasonally changing factors (body size, temperature and availability). Direct N2O emission by E. danica larvae was limited by low temperature in winter, larval development in spring and low availability in summer.

4. Both E. danica and the non‐emitting S. lutaria increased the total N2O and N2 emission from sediment in a density‐dependent manner. While N2O directly emitted by benthic invertebrates can be partially consumed in the sediment (E. danica), non‐emitting species can still indirectly contribute to total N2O emission from sediment (S. lutaria).
Differentiating aquatic plant communities in a eutrophic river using hyperspectral and multispectral remote sensing
Authors: TIAN YONG Q. , YU QIAN , ZIMMERMAN MARC J. , FLINT SUZANNE , WALDRON MARCUS C.
Published: February 2010

1. This study evaluates the efficacy of remote sensing technology to monitor species composition, areal extent and density of aquatic plants (macrophytes and filamentous algae) in impoundments where their presence may violate water‐quality standards.

2. Multispectral satellite (IKONOS) images and more than 500 in situ hyperspectral samples were acquired to map aquatic plant distributions. By analyzing field measurements, we created a library of hyperspectral signatures for a variety of aquatic plant species, associations and densities. We also used three vegetation indices. Normalized Difference Vegetation Index (NDVI), near‐infrared (NIR)‐Green Angle Index (NGAI) and normalized water absorption depth (DH), at wavelengths 554, 680, 820 and 977 nm to differentiate among aquatic plant species composition, areal density and thickness in cases where hyperspectral analysis yielded potentially ambiguous interpretations.

3. We compared the NDVI derived from IKONOS imagery with the in situ, hyperspectral‐derived NDVI. The IKONOS‐based images were also compared to data obtained through routine visual observations. Our results confirmed that aquatic species composition alters spectral signatures and affects the accuracy of remote sensing of aquatic plant density. The results also demonstrated that the NGAI has apparent advantages in estimating density over the NDVI and the DH.

4. In the feature space of the three indices, 3D scatter plot analysis revealed that hyperspectral data can differentiate several aquatic plant associations. High‐resolution multispectral imagery provided useful information to distinguish among biophysical aquatic plant characteristics. Classification analysis indicated that using satellite imagery to assess Lemna coverage yielded an overall agreement of 79% with visual observations and >90% agreement for the densest aquatic plant coverages.

5. Interpretation of biophysical parameters derived from high‐resolution satellite or airborne imagery should prove to be a valuable approach for assessing the effectiveness of management practices for controlling aquatic plant growth in inland waters, as well as for routine monitoring of aquatic plants in lakes and suitable lentic environments.
Land use and season affect fluxes of CO2, CH4, CO, N2O, H2 and isotopic source signatures in Panama: evidence from nocturnal boundary layer profiles
Authors: PENDALL E. , SCHWENDENMANN L. , RAHN T. , MILLER J. B. , TANS P. P. , WHITE J. W. C.
Published: February 2010

Conversion of tropical rainforests to pastures and plantations is associated with changes in soil properties and biogeochemical cycling, with implications for carbon cycling and trace gas fluxes. The stable isotopic composition of ecosystem respiration (δ13CR and δ18OR) is used in inversion models to quantify regional patterns of CO2 sources and sinks, but models are limited by sparse measurements in tropical regions. We measured soil respiration rates, concentrations of CO2, CH4, CO, N2O and H2 and the isotopic composition of CO2, CH4 and H2 at four heights in the nocturnal boundary layer (NBL) above three common land‐use types in central Panama, during dry and rainy seasons. Soil respiration rates were lowest in Plantation (average 3.4 μmol m−2 s−1), highest in Pasture (8.3 μmol m−2 s−1) and intermediate in Rainforest (5.2 μmol m−2 s−1). δ13CR closely reflected land use and increased during the dry season where C3 vegetation was present. δ18OR did not differ by land use but was lower during the rainy than the dry season. CO2 was correlated with other species in approximately half of the NBL profiles, allowing us to estimate trace gas fluxes that were generally within the range of literature values. The Rainforest soil was a sink for CH4 but emissions were observed in Pasture and Plantation, especially during the wet season. N2O emissions were higher in Pasture and Plantation than Rainforest, contrary to expectations. Soil H2 uptake was highest in Rainforest and was not observable in Pasture and Plantation during the wet season. We observed soil CO uptake during the dry season and emissions during the wet season across land‐use types. This study demonstrated that strong impacts of land‐use change on soil–atmosphere trace gas exchange can be detected in the NBL, and provides useful observational constraints for top‐down and bottom‐up biogeochemistry models.
Mycobacterium tuberculosis conserved hypothetical protein rRv2626c modulates macrophage effector functions
Authors: Bashir Nasreena , Kounsar Fozia , Mukhopadhyay Sangita , Hasnain Seyed E.
Published: February 2010

Secretory proteins of Mycobacterium tuberculosis are the major immunomodulators of the host immune response. Open reading frame (ORF) Rv2626c, encoding a conserved hypothetical protein eliciting a strong humoral immune response in patients with tuberculosis (TB), was shown to be up‐regulated upon infection in mice under hypoxic conditions. We now show that recombinant Rv2626c protein (rRv2626c) can bind to the surface of murine macrophages and elicit the type‐1 immune response, as manifested by nitric oxide (NO) secretion and expression of inducible nitric oxide synthase (iNOS). Significant induction of pro‐inflammatory cytokines [interleukin (IL)‐12 and tumour necrosis factor (TNF)‐α] was evident upon stimulation of murine macrophages, as well as peripheral blood mononuclear cells (PBMCs) isolated from patients with active TB disease, with rRv2626c. Stimulation with rRv2626c also enhanced the expression of costimulatory molecules such as B7‐1, B7‐2 and CD40 on murine macrophages. We further show that the production of NO and pro‐inflammatory cytokines in response to rRv2626c is mediated by the transcription factor nuclear factor (NF)‐κB, and this was further confirmed using pyrrolidine dithiocarbamate (PDTC), a specific pharmacological inhibitor of NF‐κB. Rv2626c therefore appears to modulate macrophage effector functions by eliciting both innate and adaptive immune responses, suggesting its possible use as a vaccine candidate.
Enhancement of humoral and cellular immunity with an anti‐glucocorticoid‐induced tumour necrosis factor receptor monoclonal antibody
Authors: Ponte Jose F. , Ponath Paul , Gulati Reema , Slavonic Michael , Paglia Michael , O’Shea Adam , Tone Masahide , Waldmann Herman , Vaickus Louis , Rosenzweig Michael
Published: February 2010

Adjuvants, including antibodies to tumour necrosis factor receptor superfamily members, augment immune responses. One member of this family, glucocorticoid‐induced tumour necrosis factor receptor (GITR), is expressed at low levels on naive/resting T cells, B cells and macrophages, but at higher levels on T regulatory cells. The aim of this study was to determine the ability of a rat anti‐mouse GITR monoclonal antibody, 2F8, to stimulate murine humoral and cellular immunity in a prime boost model with particular attention to posology and antigen‐specific effects. 2F8 enhanced the humoral immune response to ovalbumin and haemagglutinin (HA) compared with controls and this enhancement was equal to or greater than that obtained in mice dosed with standard adjuvants. 2F8 F(ab′)2 fragments were as effective as intact antibody in boosting humoral immunity, indicating that FcR‐mediated cross‐linking of 2F8 is not required for efficacy. Moreover, the enhanced response was durable and antigen specific. Administration of 2F8 shifted the immune response towards a T helper type 1 response with significant enhancement of immunoglobulin G2a‐ and G2b‐specific anti‐HA antibodies, as well as enhanced cellular immunity as measured by ELISPOT. 2F8‐treated mice also generated significantly more neutralizing antibodies to HA than control mice. Our findings show that anti‐GITR is a robust, versatile adjuvant that, unlike commonly used adjuvants that primarily enhance humoral immunity, enhances both humoral and cellular immunity. These results support the continued development of anti‐GITR for such indications as haematological and solid tumours, chronic viral infections, and as a vaccine adjuvant.