Accounting for the increasing benefits from scarce ecosystems.

As people get richer, and ecosystem services scarcer, policy-relevant estimates of ecosystem value must rise.

Network analysis of depression and anxiety symptom relationships in a psychiatric sample.

BACKGROUND: Researchers have studied psychological disorders extensively from a common cause perspective, in which symptoms are treated as independent indicators of an underlying disease. In contrast, the causal systems perspective seeks to understand the importance of individual symptoms and symptom-to-symptom relationships. In the current study, we used network analysis to examine the relationships between and among depression and anxiety symptoms from the causal systems perspective. METHOD: We utilized data from a large psychiatric sample at admission and discharge from a partial hospital program (N = 1029, mean treatment duration = 8 days). We investigated features of the depression/anxiety network including topology, network centrality, stability of the network at admission and discharge, as well as change in the network over the course of treatment. RESULTS: Individual symptoms of depression and anxiety were more related to other symptoms within each disorder than to symptoms between disorders. Sad mood and worry were among the most central symptoms in the network. The network structure was stable both at admission and between admission and discharge, although the overall strength of symptom relationships increased as symptom severity decreased over the course of treatment. CONCLUSIONS: Examining depression and anxiety symptoms as dynamic systems may provide novel insights into the maintenance of these mental health problems.

A variation of the Nijmegen-Bethesda assay using heat or a novel heat/cold pretreatment for the detection of FIX inhibitors in the presence of residual FIX activity.

INTRODUCTION: The inclusion of a heat treatment step has improved the classic Nijmegen-Bethesda assay for detection of factor VIII (FVIII) inhibitors in the presence of residual FVIII activity (FVIII:C). However, information regarding heat-modified Nijmegen-Bethesda assays for the detection of FIX inhibitors is still limited. METHODS: Three methods to measure FIX inhibitors in the presence or absence of residual FIX activity (FIX:C) using three different activated partial thromboplastin time (aPTT) reagents were investigated. These included the standard Nijmegen-Bethesda assay (method 1), a heat-modified assay (method 2) and a novel heat/cold-modified assay (method 3). RESULTS: In the absence of FIX:C, all methods measured similar levels of FIX inhibitor, while FIX inhibitor titres varied widely in the presence of residual FIX:C. Using method 1, inhibitors were not accurately measured in the presence of residual circulating FIX:C. Using method 2, detection was improved, especially at higher inhibitor titres. Using method 3, some additional sensitivity was obtained. The choice of aPTT reagent did not affect the detection of inhibitors. CONCLUSION: Heat pretreatment is recommended for detecting FIX inhibitors in samples with residual FIX:C. The heat/cold modification improved the sensitivity of the Nijmegen-Bethesda assay, resulting in higher tolerance for residual FIX:C.

Elevated amygdala response to faces following early deprivation.

A functional neuroimaging study examined the long-term neural correlates of early adverse rearing conditions in humans as they relate to socio-emotional development. Previously institutionalized (PI) children and a same-aged comparison group were scanned using functional magnetic resonance imaging (fMRI) while performing an Emotional Face Go/Nogo task. PI children showed heightened activity of the amygdala, a region that supports emotional learning and reactivity to emotional stimuli, and corresponding decreases in cortical regions that support perceptual and cognitive functions. Amygdala activity was associated with decreased eye-contact as measured by eye-tracking methods and during a live dyadic interaction. The association between early rearing environment and subsequent eye-contact was mediated by amygdala activity. These data support the hypothesis that early adversity alters human brain development in a way that can persist into childhood, and they offer insight into the socio-emotional disturbances in human behavior following early adversity.

The T cell response to major grass allergens is regulated and includes IL-10 production in atopic but not in non-atopic subjects.

BACKGROUND: The incidence of allergic diseases is increasing in industrialized countries and the immunological mechanisms leading to tolerance or allergy are poorly understood. Cytokines with suppressive abilities and CD4(+)CD25(+) regulatory T cells have been suggested to play a central role in allergen-specific responses. The aim was to determine whether major grass allergens induce production of suppressive cytokines in allergic and healthy subjects and to examine the inhibitory effect of these cytokines on allergic responses. METHODS: Peripheral blood mononuclear cells (PBMCs) were isolated from healthy and grass-allergic donors and stimulated with the major grass allergens Phl p 1 or Phl p 5. The effects of endogenous IL-10 and/or TGF-beta on proliferation and cytokine production were determined by use of blocking antibodies. In addition, the number of CD4(+)CD25(+) T cells and their expression of chemokine receptors were investigated by flow cytometry. RESULTS: Phl p 1 and Phl p 5 induced IL-10 production, which down-regulated proliferation and cytokine production, in PBMC cultures from atopic but not from non-atopic donors. Comparable frequencies of CD4(+)CD25(+) T cells were present in PBMCs in the two groups, but fewer cells from atopic donors were CD4(+)CD25(+)CCR4(+) and more cells were CD4(+)CD25(+)CLA(+) compared to healthy donors. CONCLUSION: Allergen-specific responses of grass allergic patients but not in non-atopic subjects are influenced by regulatory cytokines produced in response to the important allergens. Differences in CD4(+)CD25(+) T cell expression of chemokine receptors in allergic compared to non-atopic donors could suggest that the homing of CD4(+)CD25(+) T cells is important for the regulation of allergen-specific responses.

In vitro effects of monophthalates on cytokine expression in the monocytic cell line THP-1 and in peripheral blood mononuclear cells from allergic and non-allergic donors.

It has recently been shown that plasticizers are present in indoor air dust, which may lead to human exposure via the inhalation route. Moreover, studies have indicated that plasticizers may possess adjuvant effects increasing the health damaging potential of allergens. The aim of this study was to investigate the in vitro effect of metabolites of phthalate plastisizers, such as whether an adjuvant effect is paralleled by changes of the cytokine expression in the monocytic cell line THP-1 and in peripheral blood mononuclear cells (PBMCs) from allergics and non-allergics. The toxicity monitored by cell viability was determined by incubating THP-1 cells with a 10-fold dilution series of monophthalates for 24 h. At different points in time cytokine expression (IL-1beta, IL-6, IL-12alpha (p35)) in THP-1 cells incubated with non-toxic concentrations of monophthalate (2-20 microg/ml)+/-LPS (1 microg/ml) were determined using Quantitative Competitive RT-PCR. PBMCs from allergics and non-allergics were incubated with monophthalate 220 microg/ml) for up to 48 h and cytokine expression (IL-4, IL-5, IFN-gamma) was measured using real-time PCR. The cytotoxic level of monophthalates is 20-200 microg/ml, depending on the individual monophthalate. There seems to be a correlation between increasing side-chain length and toxicity. Monophthalates did not induce changes in cytokine expression in THP-1 cells, though there is an increase when co-incubating with LPS. Cytokine expression in PBMC seems virtually unchanged when co-incubated with monophthalate, though mono-n-butyl phthalate (MBUP) tends to increase the level of IL-4 in PBMCs from allergic individuals. The two cellular models demonstrated the dynamics of regulated cytokine mRNA and are applicable for in vitro immunotoxicological investigations. The results regarding monophthalates suggest these to have a limited effect on cytokine expression in the monocytic cell line THP-1 and weak effect on cytokine expression in PBMCs from allergic and non-allergic individuals.

Detergents in the indoor environment - what is the evidence for an allergy promoting effect? Known and postulated mechanisms.

IgE-mediated allergic diseases, such as asthma and rhinitis seem to be increasing in industrialised societies. One possible explanation for this could be the increased use of more effective and aggressive detergents. The surfactants from these could interfere with the sensitisation process in which specific IgE is formed to ubiquitously occurring environmental allergens. Only sparse data exist in relation to surfactants and allergic sensitization. However, it can be speculated that the strong surfactant properties of some of ingredients used in modem detergents may interfere with some of the intricate cellular interactions taking place along the immunological pathways. These include formation of IL-4 and IL-5 producing T helper lymphocytes type 2 and the B-lymphocyte isotype switch, which leads to production of specific IgE. Candidates for experimental studies of such phenomena on the cellular level are proposed.

CXC chemokine receptor 3 expression on CD34(+) hematopoietic progenitors from human cord blood induced by granulocyte-macrophage colony-stimulating factor: chemotaxis and adhesion induced by its ligands, interferon gamma-inducible protein 10 and monokine induced by interferon gamma.

CXC chemokine receptor 3 (CXCR3), which is known to be expressed predominately on memory and activated T lymphocytes, is a receptor for both interferon gamma (IFN-gamma)-inducible protein 10 (gamma IP-10) and monokine induced by IFN-gamma (Mig). We report the novel finding that CXCR3 is also expressed on CD34(+) hematopoietic progenitors from human cord blood stimulated with granulocyte-macrophage colony-stimulating factor (GM-CSF) but not on freshly isolated CD34(+) progenitors. Freshly isolated CD34(+) progenitors expressed low levels of CXCR3 messenger RNA, but this expression was highly up-regulated by GM-CSF, as indicated by a real-time quantitative reverse transcriptase-polymerase chain reaction technique. gamma IP-10 and Mig induced chemotaxis of GM-CSF-stimulated CD34(+) progenitors by means of CXCR3, since an anti-CXCR3 monoclonal antibody (mAb) was found to block gamma IP-10-induced and Mig-induced CD34(+) progenitor chemotaxis. These chemotactic attracted CD34(+) progenitors are colony-forming units-granulocyte-macrophage. gamma IP-10 and Mig also induced GM-CSF-stimulated CD34(+) progenitor adhesion and aggregation by means of CXCR3, a finding confirmed by the observation that anti-CXCR3 mAb blocked these functions of gammaIP-10 and Mig but not of chemokine stromal cell-derived factor 1 alpha. gamma IP-10-induced and Mig-induced up-regulation of integrins (CD49a and CD49b) was found to play a crucial role in adhesion of GM-CSF-stimulated CD34(+) progenitors. Moreover, gamma IP-10 and Mig stimulated CXCR3 redistribution and cellular polarization in GM-CSF-stimulated CD34(+) progenitors. These results indicate that CXCR3-gamma IP-10 and CXCR3-Mig receptor-ligand pairs, as well as the effects of GM-CSF on them, may be especially important in the cytokine/chemokine environment for the physiologic and pathophysiologic events of differentiation of CD34(+) hematopoietic progenitors into lymphoid and myeloid stem cells, subsequently immune and inflammatory cells. These processes include transmigration, relocation, differentiation, and maturation of CD34(+) hematopoietic progenitors. (Blood. 2000;96:1230-1238)

CXCR3 expression and activation of eosinophils: role of IFN-gamma-inducible protein-10 and monokine induced by IFN-gamma.

CXC chemokine receptor 3 (CXCR3), predominately expressed on memory/activated T lymphocytes, is a receptor for both IFN-gamma-inducible protein-10 (gamma IP-10) and monokine induced by IFN-gamma (Mig). We report a novel finding that CXCR3 is also expressed on eosinophils. gamma IP-10 and Mig induce eosinophil chemotaxis via CXCR3, as documented by the fact that anti-CXCR3 mAb blocks gamma IP-10- and Mig-induced eosinophil chemotaxis. gamma IP-10- and Mig-induced eosinophil chemotaxis are up- and down-regulated by IL-2 and IL-10, respectively. Correspondingly, CXCR3 protein and mRNA expressions in eosinophils are up- and down-regulated by IL-2 and IL-10, respectively, as detected using flow cytometry, immunocytochemical assay, and a real-time quantitative RT-PCR technique. gamma IP-10 and Mig act eosinophils to induce chemotaxis via the cAMP-dependent protein kinase A signaling pathways. The fact that gamma IP-10 and Mig induce an increase in intracellular calcium in eosinophils confirms that CXCR3 exists on eosinophils. Besides induction to chemotaxis, gamma IP-10 and Mig also activate eosinophils to eosinophil cationic protein release. These results indicate that CXCR3-gamma IP-10 and -Mig receptor-ligand pairs as well as the effects of IL-2 and IL-10 on them may be especially important in the cytokine/chemokine environment for the pathophysiologic events of allergic inflammation, including initiation, progression, and termination in the processes.

Cutting edge: expression of the NF of activated T cells in eosinophils: regulation by IL-4 and IL-5.

We report that NF-AT1 and NF-AT4 are expressed cytoplasmically in resting eosinophils, whereas NF-AT2 and NF-AT3 have not been seen. Likewise, NF-AT1 mRNA and NF-AT4 mRNA have been detected in resting eosinophils, and their levels can be significantly up-regulated by the Th2-associated cytokines IL-4 and IL-5. There is no detectable NF-AT protein expression in the nuclei of resting eosinophils. However NF-ATs appear in the nuclei of IL-4-, IL-5-, or ionomycin-stimulated eosinophils. Only NF-AT1 and NF-AT4, but not NF-AT2 and NF-AT3, have translocated into the nuclei in IL-4- or IL-5-stimulated eosinophils. These findings delineate a novel pathway in the cytokine network in which Th2 lymphocytes "control" eosinophils via the release of IL-4 and IL-5, and activation of NF-AT in eosinophils. The findings also suggest that a later feedback "talking" may exist between eosinophils and Th2 lymphocytes.

Topical bleomycin for the treatment of dysplastic oral leukoplakia.

BACKGROUND: Because malignant transformation of dysplastic oral leukoplakia has been reported in up to 43% of cases, these lesions must be managed. METHODS: This study evaluated the use of topical 1% bleomycin in dimethylsulfoxide for the treatment of dysplastic oral lesions. Bleomycin was applied once daily for 14 consecutive days to lesions of the oral mucosa in 19 patients. Immediate posttreatment biopsies and the clinical response were evaluated and clinical follow-up was conducted for as long as possible. RESULTS: The mean age of the patients at diagnosis was 59.4 years and 74% were tobacco users. Seventy-five percent of patients had resolution of dysplasia at follow-up biopsy, with a mean improvement of two histologic grades of dysplasia after topical chemotherapy. Ninety-four percent of the patients attained at least partial responses. After a mean follow-up period of 3.4 years, 31.6% of patients had no clinically visible lesions and 47.4% of patients had clinically benign lesions of homogeneous leukoplakia or minimal visible leukoplakia. In 2 patients (11%) malignant transformation occurred a mean of 1.75 years after bleomycin treatment. CONCLUSIONS: Topical bleomycin may prevent the potential progression of leukoplakia through dysplasia to carcinoma. Close follow-up of all patients with dysplasia is required.

Disruption of target DNA binding in Mu DNA transposition by alteration of position 99 in the Mu B protein.

Target DNA binding by the Mu B protein is an important step in phage Mu transposition; however, the region of Mu B involved in target binding and the mechanism of the interaction are unknown. Previous studies have demonstrated that modification of Mu B with the sulfhydryl-specific reagent N-ethylmaleimide can selectively inhibit target DNA binding. We now show that individual mutation of the three cysteines in Mu B to serine results in proteins which are active in intermolecular strand transfer, but demonstrate variable levels of N-ethylmaleimide resistance. The data indicate that cysteine 99 is the primary site of modification affecting target DNA binding, with a minor contribution resulting from the derivatization of cysteine 129. These findings are confirmed by the construction of Mu B mutants containing a bulky side-chain at the individual cysteine to mimic the N-ethylmaleimide modified protein. The C99Y protein shows a complete loss in target-dependent strand transfer activity under standard reaction conditions and C129Y displays partial activity. The effect of the tyrosine substitutions is specific for target interaction as both mutants show wild-type activity in their ability to stimulate the Mu transposase to perform donor cleavage and intramolecular strand transfer. Finally, a target dissociation assay has shown that the C99Y-DNA complex generated in the presence of ATP-gamma-S has a drastically reduced half-life as previously found for N-ethylmaleimide treated wild-type Mu B. Modification of cysteine 99 is proposed to block target DNA binding by causing steric interference near the DNA binding pocket.

The utility of toluidine blue application as a diagnostic aid in patients previously treated for upper oropharyngeal carcinoma.

OBJECTIVES: This study evaluated the utility (usefulness) of toluidine blue application as an aid to the recognition and diagnosis of clinically evident lesions in a series of patients previously treated for oral cancer and monitored in a cancer center. In addition to increased risk of recurrence of cancer or new second primary lesions, patients who have had previous treatment for oropharyngeal cancer may be more difficult to assess because of tissue changes that occur as a result of previous radiation therapy. STUDY DESIGN: Patients with a history of oral malignancy were assessed by clinical examination followed by application of toluidine blue. Biopsy sites were determined on the basis of unaided visual examination and by the findings on toluidine blue application. Biopsy specimens were reviewed by a pathologist blinded to the clinical findings. RESULTS: Unaided clinical examination identified 78% of carcinoma in situ or invasive malignant lesions compared with toluidine blue application, which identified all (100%) carcinoma in situ or invasive malignant lesions (p = 0.02) and produced no false-negative findings. No differences were found between clinical examination and toluidine application in the detection of dysplastic lesions. CONCLUSION: Toluidine blue retention was seen in all cases of carcinoma in situ and invasive carcinoma, and no false-negative findings were seen with toluidine blue. When used by a trained and experienced clinician in a cancer center, toluidine blue was a valuable visual aid to clinical examination of oral mucosal lesions.

Anatomy of a flexer-DNA complex inside a higher-order transposition intermediate.

SUMMARY: Escherichia coli HU, a nonsequence-specific histone- and HMG-like DNA-binding protein, was chemically converted into a series of HU-nucleases with an iron-EDTA-based cleavage moiety positioned at 16 rationally selected sites. Specific DNA cleavage patterns from each of these HU-nucleases allowed us to determine the precise localization, stoichiometry, and orientation of HU binding in the Mu transpososome, a multiprotein structure that mediates the chemical reactions in DNA transposition. Correlation of the DNA cleavage data with the position of the cleavage moiety in the HU three-dimensional structure indicates the presence of a dramatic DNA bend, for which the bend center, direction, and magnitude were assessed. The data, which directly localize selected HU amino acids with respect to DNA in the transpososome, were used as constraints for computer-based molecular modeling to derive the first snapshot of an HU-DNA interaction.

Topical bleomycin treatment of oral leukoplakia: a randomized double-blind clinical trial.

BACKGROUND: Oral leukoplakia and oral erythroplakia may be associated with benign and dysplastic cellular changes, and are at risk of malignant transformation. Additional means of management of these lesions is needed. The results of nonblinded trials using topical bleomycin in oral leukoplakia indicated the need for phase III study. METHODS: A prospective, double-blind, randomized trial of topical bleomycin versus placebo was conducted. Bleomycin 1% in dimethylsulphoxide (DMSO) or the carrier was applied for 5 minutes for 14 consecutive days. Clinical assessment and pre-application and post-treatment biopsies were conducted. RESULTS: Twenty-two patients were randomized. Of the patients who received bleomycin, decrease in clinical size of the lesion was achieved (p = 0.001), and histological reduction in dysplasia was seen (p = 0.094). CONCLUSIONS: The topical application of bleomycin in DMSO may represent an additional approach to management of oral leukoplakia. The treatment is well-tolerated, and may be considered when the location or extent of the lesion may make surgical excision difficult.

Development of an efficient electrohydraulic total artificial heart.

The efficient use of space and energy is achieved in a new implantable total artificial heart (TAH). To fit the orthotopic thoracic space, toroidal blood pumps encircle an energy converter, consisting of a centrifugal hydraulic fluid pump and a rotary reversing valve. The new centrifugal pump produces cardiac outputs of 6 L/min, with an average hydraulic efficiency of 47%. Toroidal blood pumps encircling the pump deliver a 60 cc stroke with an 85% ejection fraction and have a predictable blood flow pattern with no stagnant regions. Components have been characterized in vitro, and blood contacting elements have been tested in vivo.

Treatment of oral leukoplakia with topical bleomycin. A pilot study.

Leukoplakia is a clinical diagnosis with the potential for malignant transformation. This pilot study tested the use of bleomycin a as topical agent for the treatment of leukoplakia. Twelve patients were enrolled in this study, each for 2 weeks of daily topical application of bleomycin in dimethylsulphoxide (DMSO) painted over the lesion. Two patients were excluded from analysis due to excessive dilution of the medication by local saliva production. Five patients were treated with a 0.5% solution that caused a decrease in the thickness of all of the lesions clinically. One lesion also had a sharp decrease in size. Five patients were treated with a 1.0% solution that caused a complete resolution of the lesions in three patients. Two lesions demonstrated maturation of parakeratosis to keratosis only. All patients tolerated the treatments well. Topical bleomycin may play a role in the treatment of leukoplakia and warrants further investigation.

Organisation and results of the cervical cytology screening programme in British Columbia, 1955-85.

A screening programme to detect preinvasive carcinoma of the cervix was started in British Columbia in 1949. Since 1970 the number of women who have been screened at least once has been maintained at about 85% of the population at risk. More than 500,000 cervical smears are being examined each year in the central laboratory. There has been an appreciable increase in the number of cases and rates of carcinoma in situ seen since 1970, particularly in women between 20 and 30 years of age. Since the programme started over 26,000 cases of squamous carcinoma in situ have been detected and treated. The incidence of clinically invasive squamous carcinoma of the cervix has fallen by 78% during the period under review, and mortality from squamous carcinoma of the cervix has fallen by 72%. A colposcopy programme, introduced throughout British Columbia over the past 12 years, has been important in reducing the problems of managing preinvasive lesions, particularly in younger women. It is concluded that the reduction in morbidity and mortality from invasive squamous cancer of the cervix in British Columbia over the past 30 years is directly attributable to the province wide screening programme and that a large potential increase in invasive cervical cancer rates among younger women is being prevented.

Colposcopic evaluation of pregnant patients with abnormal cervical smears.

Colposcopic examination and biopsy were used to assess 123 pregnant patients presenting with abnormal cervical smears. Eighty-seven per cent were 30 years of age or less and 95 (77 per cent) had had one or no previous children. Two patients were found to have microinvasive carcinoma and, in an additional 95 patients, either severe dysplasia or carcinoma in situ was present. Fifty-five patients (45 per cent) had subsequent conization or hysterectomy and in no instance was the histological diagnosis more serious than that anticipated from the colposcopic evaluation. Only three patients (1-6 per cent) had a cone biopsy during pregnancy; only one minor complication occurred. Colposcopic examination is the choice method of evaluating patients with abnormal cervical smears in pregnancy.