The Communicative-Relational Operating Model of the Italian National Institute of Health for an Effective Telephone Intervention in Public Health, Structured on Basic Counselling Skills.

Introduction: Communication has a crucial role in public health, because it becomes an essential component of prevention; it is also a proactive tool in health promotion. From a planning perspective, it is appropriate to use communication means that can help the bidirectional communication process, such as face-to-face communication and telephone communication. Materials and methods: In relation to this, the Italian National Institute of Health has developed the "Modello Operativo Comunicativo-Relazionale" (the "Communicative-Relational Operating Model"). It is based on the fundamental skills of the counselling, this gives a protocol to the health professionals that is replicable and organized and it allows health professionals to carry out a telephone communication that is efficient with the user through technical-scientific and communication-relational skills. The goal is to answer in a customized way to the various users' health needs. The Operating Model was created by experts of the National AIDS and Sexually Transmitted Infections Helpline of the Operational Unit of Psycho-Socio-Behavioural Research, Communication, Training, of the Infectious Diseases Department. Later, the Operating Model was proposed to the experts of the Helplines in the National Centre on Addictions and Doping and the National Helpline of the National Centre for Rare Diseases in the National Institute of Health that integrated this method into their telephone approach. Results: The Operating Model illustrated above was applied to several helplines of the National Institute of Health as an example of correct scientific information, updated and customized on sexual transmitted infections, addictions and rare diseases. Conclusions: This article aims to illustrate the Operating Model, the theoretical prerequisites that subtend it and its possible application in the different public health structures that use the telephone for a profes-sional relationship with their users.

Identification of Vibrio alginolyticus virulent strain-specific DNA regions by suppression subtractive hybridization and PCR.

AIMS: Vibrio alginolyticus was frequently isolated from diseased farmed fish in the coaster waters of Hainan Island over the past two decades. In this study, we attempted to identify candidates of virulent strain-specific DNA regions for this pathogen. METHODS AND RESULTS: Suppression subtractive hybridization (SSH) and PCR were successively performed between the typical virulent strain and avirulent strain of V. alginolyticus, in which they shared 99·54% homology of 16S rDNAs. Out of 2873 subtracted clones, nine clones were finally indicated to harbour virulent strain-specific DNA fragments. The receivable functions of the major fragments in the nine clones were believed to encode methyl-accepting chemotaxis protein (n = 1), type VI secretion system-associated FHA domain protein TagH (n = 1), diguanylate cyclase (n = 1), AraC family transcriptional regulator (n = 1), ABC-type uncharacterized transport system permease component (n = 1) and hypothetical proteins (n = 4). Two hypothetical proteins contain several disordered regions. CONCLUSIONS: Some specific DNA regions existed in the virulent strain of V. alginolyticus, and the SSH assay could be a highly sensitive method for identifying virulent regions in pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: This report is the first to describe the identification of virulent strain-specific DNA regions in the V. alginolyticus genome, which is helpful in developing virulent strain-specific rapid detection methods and is a pivotal precondition for clarifying the molecular virulence mechanism of V. alginolyticus.

Super-factors associated with transmission of occupational COVID-19 infection among healthcare staff in Wuhan, China.

BACKGROUND: Globally, there have been many cases of coronavirus disease 2019 (COVID-19) among medical staff; however, the main factors associated with the infection are not well understood. AIM: To identify the super-factors causing COVID-19 infection in medical staff in China. METHODS: A cross-sectional study was conducted between January 1st and February 30th, 2020, in which front-line members of medical staff who took part in the care and treatment of patients with COVID-19 were enrolled. Epidemiological and demographic data between infected and uninfected groups were collected and compared. Social network analysis (SNA) was used to establish socio-metric social links between influencing factors. FINDINGS: A total of 92 medical staff were enrolled. In all participant groups, the super-factor identified by the network was wearing a medical protective mask or surgical mask correctly (degree: 572; closeness: 25; betweenness centrality: 3.23). Touching the cheek, nose, and mouth while working was the super-factor in the infected group. This was the biggest node in the network and had the strongest influence (degree: 370; closeness: 29; betweenness centrality: 0.37). Self-protection score was the super-factor in the uninfected group but was the isolated factor in the infected group (degree: 201; closeness: 28; betweenness centrality: 5.64). For family members, the exposure history to Huanan Seafood Wholesale Market and the contact history to wild animals were two isolated nodes. CONCLUSION: High self-protection score was the main factor that prevented medical staff from contracting COVID-19 infection. The main factor contributing to COVID-19 infections among medical staff was touching the cheek, nose, and mouth while working.

Germline investigation in male breast cancer of DNA repair genes by next-generation sequencing.

PURPOSE: In order to better define the breast cancer (BC) genetic risk factors in men, a germline investigation was carried out on 81 Male BC cases by screening the 24 genes involved in BC predisposition, genome stability maintenance and DNA repair mechanisms by next-generation sequencing. METHODS: Germline DNAs were tested in a custom multi-gene panel focused on all coding exons and exon-intron boundaries of 24 selected genes using two amplicon-based assays on PGM-Ion Torrent (ThermoFisher Scientific) and MiSeq (Illumina) platforms. All variants were recorded and classified by using a custom pipeline. RESULTS: Clinical pathological data and the family history of 81 Male BC cases were gathered and analysed, revealing the average age of onset to be 61.3 years old and that in 35 cases there was a family history of BC. Our genetic screening allowed us to identify a germline mutation in 22 patients (23%) in 4 genes: BRCA2, BRIP1, MUTYH and PMS2. Moreover, 12 variants of unknown clinical significance (VUS) in 9 genes (BARD1, BRCA1, BRIP1, CHEK2, ERCC1, NBN, PALB2, PMS1, RAD50) were predicted as potentially pathogenic by in silico analysis bringing the mutation detection rate up to 40%. CONCLUSION: As expected, a positive family history is a strong predictor of germline BRCA2 mutations in male BC. Understanding the potential pathogenicity of VUS represents an extremely urgent need for the management of BC risk in Male BC cases and their own families.

A multicentre randomised controlled trial of the effect of intra-operative dexmedetomidine on cognitive decline after surgery.

Peri-operative dexmedetomidine can reduce rates of delirium immediately after surgery. We aimed to assess the effect of dexmedetomidine on cognition up to six postoperative months and its association with changes in serum concentrations of brain-derived neurotrophic factor on the third and seventh postoperative days. We randomly allocated 535 patients aged 65 years or more undergoing scheduled gastro-intestinal laparotomy to: intra-operative dexmedetomidine, 0.5 µg.kg-1 bolus followed by 0.4 µg.kg-1 .hr-1 infusion (n = 269), or placebo (n = 266). Dexmedetomidine reduced the rate of cognitive impairment: on the third postoperative day, 40/269 vs. 65/266, p = 0.006; on the seventh postoperative day, 31/269 vs. 49/266, p = 0.03 and at one postoperative month, 42/250 vs. 61/248, p = 0.04. Cognitive impairment at seven postoperative days was associated with changes in brain-derived neurotrophic factor concentrations on the third and seventh postoperative days; area under the receiver operating characteristic curve 0.63, p < 0.001 and 0.58, p = 0.016, respectively. Intra-operative dexmedetomidine reduced cognitive decline up to one postoperative month in elderly patients undergoing scheduled laparotomy, which was associated with changes in serum brain-derived neurotrophic factor.

Long non-coding RNA expression profile in permanent atrial fibrillation patients with rheumatic heart disease.

OBJECTIVE: Atrial fibrillation (AF) is the most common type of arrhythmia, especially in rheumatic heart disease (RHD) patients. The differences in structural remodeling and electrical remodeling between the left and right atrium associated with AF in RHD patients are well known, and alterations in the expression profiles of long noncoding RNAs (lncRNAs) in the left atrium have also been investigated. However, the role of lncRNAs in the right atrium (RA) remains largely unknown. PATIENTS AND METHODS: We identified differentially expressed lncRNAs in RA tissues of RHD patients with AF or a normal sinus rhythm (NSR) using microarray analysis. Then, we performed gene ontology (GO) and KEGG pathway analyses for functional annotation of the deregulated lncRNAs. Finally, we constructed a lncRNA-mRNA co-expression network. RESULTS: Of the 22,829 human non-coding RNAs analyzed, a total of 1,909 long non-coding RNAs were detected. A total of 182 lncRNAs (117 downregulated and 65 upregulated) were shown to be differentially expressed (fold-change > 1.5) in AF patients compared with NSR patients. Many lncRNAs might be partially involved in an AF-related pathway. CONCLUSIONS: AF dysregulates the expression of lncRNAs in the RA of RHD patients. These findings may be useful for exploring potential therapeutic treatments for AF in RHD patients.

Changes in health-related quality of life with treatment of longer-acting clotting factors: results in the A-LONG and B-LONG clinical studies.

INTRODUCTION: In haemophilia, prophylactic infusion of replacement factor can result in improvements in health-related quality of life (HRQoL) when compared with episodic treatment. The Haemophilia-specific Quality of Life (Haem-A-QoL) questionnaire assessed HRQoL in adults with severe haemophilia A or B who received prophylactic or episodic treatment with recombinant factor VIII or IX Fc fusion protein (rFVIIIFc or rFIXFc) in the A-LONG or B-LONG clinical studies. AIMS: Understand changes in HRQoL during the A-LONG and B-LONG trials. METHODS: Group-level and individual-level changes over time for the Haem-A-QoL key domains of 'Physical Health' and 'Sports & Leisure,' and 'Total Score' were evaluated in adults through baseline and 6-month HRQoL assessments. Previously determined responder definitions (RDs) were used for evaluating meaningful subject-level HRQoL improvements. RESULTS: The analysis included 67 A-LONG and 51 B-LONG subjects who completed the Haem-A-QoL (baseline and 6 months). While HRQoL improvements were observed among all treatment groups, greater improvements in HRQoL were observed among subjects who received episodic treatment pre-study (and prophylaxis on-study) compared to those who received hyphenate prophylaxis. Applying the RDs for interpreting 6-month changes, 47.4%/33.3% ('Physical Health'), 35.9%/50.0% ('Sports & Leisure') and 23.9%/33.3% ('Total Score') of A-LONG subjects who received individualized or weekly prophylaxis were classified as HRQoL responders. In B-LONG, 69.2%/57.9% ('Physical Health'), 44.4%/56.7% ('Sports & Leisure') and 41.7%/44.1% ('Total Score') of subjects who received individualized or weekly prophylaxis were classified as HRQoL responders. CONCLUSION: Changes in Haem-A-QoL key domains and 'Total Score' suggest that prophylaxis with long-acting rFVIIIFc or rFIXFc resulted in meaningful HRQoL improvements.

Validation of the manufacturing process used to produce long-acting recombinant factor IX Fc fusion protein.

Recombinant factor IX Fc (rFIXFc) fusion protein is the first of a new class of bioengineered long-acting factors approved for the treatment and prevention of bleeding episodes in haemophilia B. The aim of this work was to describe the manufacturing process for rFIXFc, to assess product quality and to evaluate the capacity of the process to remove impurities and viruses. This manufacturing process utilized a transferable and scalable platform approach established for therapeutic antibody manufacturing and adapted for production of the rFIXFc molecule. rFIXFc was produced using a process free of human- and animal-derived raw materials and a host cell line derived from human embryonic kidney (HEK) 293H cells. The process employed multi-step purification and viral clearance processing, including use of a protein A affinity capture chromatography step, which binds to the Fc portion of the rFIXFc molecule with high affinity and specificity, and a 15 nm pore size virus removal nanofilter. Process validation studies were performed to evaluate identity, purity, activity and safety. The manufacturing process produced rFIXFc with consistent product quality and high purity. Impurity clearance validation studies demonstrated robust and reproducible removal of process-related impurities and adventitious viruses. The rFIXFc manufacturing process produces a highly pure product, free of non-human glycan structures. Validation studies demonstrate that this product is produced with consistent quality and purity. In addition, the scalability and transferability of this process are key attributes to ensure consistent and continuous supply of rFIXFc.

Ibuprofen partially attenuates neurodegenerative symptoms in presenilin conditional double-knockout mice.

Ibuprofen is a widely used nonsteroidal anti-inflammatory drug that reportedly reduces the risk of Alzheimer's disease (AD) development. The anti-inflammatory effect of ibuprofen occurred via inhibition of cyclooxygenases and anti-amyloidogenesis through modulation of γ-secretase. Presenilin 1 and 2 conditional double-knockout (cDKO) mice exhibited age-dependent memory impairment and forebrain degeneration without elevation of amyloid ß deposition. Therefore, cDKO mice can be an ideal animal model on which to independently test the effects of ibuprofen anti-inflammatory properties on the prevention of AD. Three- and six-month-old cDKO mice were fed diet containing 375 ppm ibuprofen for six months. After multiple, well-validated behavioral tests, treatment with ibuprofen improved cognition-related behavioral performance, and drug efficacy was correlated with the timing of administration. Ibuprofen was more effective on six-month-old than on three-month-old cDKO mice. Biochemical analysis demonstrated that the effects of ibuprofen on glial fibrillary acidic protein and CD68 expression levels were uneven in different brain regions of cDKO mice and that age also influenced such effects. Tau hyperphosphorylation and the cleavage of caspase-3 decreased after ibuprofen treatment, and this effect was more significant in the older than the younger group of mice, which was consistent with the results of behavioral tests.

Comparative gene expression profiling reveals key pathways in septic skeletal muscle.

AIM: Skeletal muscle transcriptome of patients with sepsis was compared with that of controls to elucidate the molecular mechanisms underlying sepsis-induced skeletal muscle dysfunction. MATERIALS AND METHODS: Gene expression data set GSE13205 was downloaded from Gene Expression Omnibus (GEO), including 13 septic samples and 8 controls. Differentially expressed genes (DEGs) were screened out with t-test. Transcriptional regulatory network was constructed for the DEGs with information from UCSU. In order to identify altered biological functions in sepsis, pathway enrichment analysis was conducted for all the genes in the network with DAVID. Besides, relevant small molecules were retrieved using the Connectivity Map (camp). RESULTS: A total of 287 DEGs were obtained in sepsis, 149 up-regulated and 138 down-regulated. A transcriptional regulatory network containing 83 nodes and 98 edges was then constructed. Five transcription factors (TFs) and their target genes were acquired. Significantly altered biological pathways included insulin signaling pathway, neurotrophin signaling pathway, fructose and mannose metabolism, circadian rhythm and apoptosis. Besides, a number of relevant molecules were obtained, such as trazodone and thapsigargin. CONCLUSIONS: Our study provided an insight into the molecular changes sepsis and related skeletal muscle dysfunction. The information could be beneficial in disclosing the pathogenesis and developing effective therapies.

Mass measurements of the neutron-deficient 41Ti, 45Cr, 49Fe, and 53Ni nuclides: first test of the isobaric multiplet mass equation in f p-shell nuclei.

Isochronous mass spectrometry has been applied to neutron-deficient 58Ni projectile fragments at the HIRFL-CSR facility in Lanzhou, China. Masses of a series of short-lived T(z)=-3/2 nuclides including 41Ti, 45Cr, 49Fe, and 53Ni have been measured with a precision of 20-40 keV. The new data enable us to test for the first time the isobaric multiplet mass equation (IMME) in fp-shell nuclei. We observe that the IMME is inconsistent with the generally accepted quadratic form for the A=53, T=3/2 quartet. We perform full space shell model calculations and compare them with the new experimental results.

Positive association of major histocompatibility complex class I chain-related gene A polymorphism with leukemia susceptibility in the people of Han nationality of Southern China.

To assess the potential contribution of major histocompatibility complex class I chain-related gene A (MICA) polymorphisms toward the pathogenesis of leukemia, 107 leukemia patients and 162 ethnically matched controls from Hunan province, Southern China, were genotyped for the MICA polymorphism using polymerase chain reaction-sequence-specific priming (PCR-SSP) and sequence-based typing (PCR-SBT). The relevance between these genotypes and risk of leukemia was assessed by means of odds ratio (OR) with 95% confidence intervals (95% CIs). Allele frequencies of MICA-sequence and MICA-STR were different in leukemia patients in comparison with normal controls (both P < 0.05). MICA A5 was directly associated with leukemia (OR = 2.3257, Pc < 0.0005), whereas MICA A5.1 and MICA*008 were inversely associated with leukemia (OR = 0.5874, Pc = 0.0235 and OR = 0.5874, Pc = 0.0329, respectively). In addition, we found that homozygotes for MICA A5 (OR = 14.0659, 95% CI: 3.1627-62.5574, Pc < 0.0001) and MICA*010 (OR = 10.1053, 95% CI: 2.2139-46.1260, Pc < 0.0004) were at an increased risk for leukemia, whereas heterozygotes for MICA*008 and MICA A5.1 were linked to a decreased risk for leukemia (OR = 0.4609, 95% CI: 0.2799-0.7590, Pc = 0.0027). MICA allelic variation is associated with leukemia in Hunan Han population; the data also suggest that MICA gene polymorphism affects susceptibility to different clinical subtypes of leukemia.

Direct mass measurements of short-lived A=2Z-1 nuclides (63)Ge, (65)As, (67)Se, and (71)Kr and their impact on nucleosynthesis in the rp process.

Mass excesses of short-lived A=2Z-1 nuclei (63)Ge, (65)As, (67)Se, and (71)Kr have been directly measured to be -46,921(37), -46,937(85), -46,580(67), and -46,320(141) keV, respectively. The deduced proton separation energy of -90(85) keV for (65)As shows that this nucleus is only slightly proton unbound. X-ray burst model calculations with the new mass excess of (65)As suggest that the majority of the reaction flow passes through (64)Ge via proton capture, indicating that (64)Ge is not a significant rp-process waiting point.

Artemin is oncogenic for human mammary carcinoma cells.

We report that artemin, a member of the glial cell line-derived neurotrophic factor family of ligands, is oncogenic for human mammary carcinoma. Artemin is expressed in numerous human mammary carcinoma cell lines. Forced expression of artemin in mammary carcinoma cells results in increased anchorage-independent growth, increased colony formation in soft agar and in three-dimensional Matrigel, and also promotes a scattered cell phenotype with enhanced migration and invasion. Moreover, forced expression of artemin increases tumor size in xenograft models and leads to highly proliferative, poorly differentiated and invasive tumors. Expression data in Oncomine indicate that high artemin expression is significantly associated with residual disease after chemotherapy, metastasis, relapse and death. Artemin protein is detectable in 65% of mammary carcinoma and its expression correlates to decreased overall survival in the cohort of patients. Depletion of endogenous artemin with small interfering RNA, or antibody inhibition of artemin, decreases the oncogenicity and invasiveness of mammary carcinoma cells. Artemin is therefore oncogenic for human mammary carcinoma, and targeted therapeutic approaches to inhibit artemin function in mammary carcinoma warrant consideration.

[Microbiological and production characteristics of the high-temperature Kongdian bed revealed during field trial of biotechnology for the enhancement of oil recovery].

Microbiological technology for the enhancement of oil recovery based on the activation of the stratal microflora was tested in the high-temperature horizons of the Kongdian bed (60 degrees C) of the Dagang oil field (China). This biotechnology consists in the pumping of a water-air mixture and nitrogen and phosphorus mineral salts into the oil stratum through injection wells in order to stimulate the activity of the stratal microflora which produce oil-releasing metabolites. Monitoring of the physicochemical, microbiological, and production characteristics of the test site has revealed large changes in the ecosystem as a result of the application of biotechnology. The cell numbers of thermophilic hydrocarbon-oxidizing, fermentative, sulfate-reducing, and methanogenic microorganisms increased 10-10 000-fold. The rates of methanogenesis and sulfate reduction increased in the near-bottom zone of the injection wells and of some production wells. The microbial oil transformation was accompanied by the accumulation of bicarbonate ions, volatile fatty acids, and biosurfactants in the formation waters, as well as of CH4 and CO2 both in the gas phase and in the oil. Microbial metabolites promoted the additional recovery of oil. As a result of the application of biotechnology, the water content in the production liquid from the test site decreased, and the oil content increased. This allowed the recovery of more than 14000 tons of additional oil over 3.5 years.

Versatile expression system for rapid and stable production of recombinant proteins.

Previously we reported the development of a novel expression system with Tat/TAR-oriP vectors and HKB11 cell line, which supports high level protein expression (Cho et al. Cytotechnology 2001, 37, 23-30). In the present study, we further demonstrated that HKB11 cells are suitable for high throughput expression (microgram scale) of genomic candidates in transient transfection system for in vitro evaluation of biological functions. HKB11 cells were also shown to support the production of milligram to gram quantities of protein drug candidates for in vivo evaluation of efficacy in various disease models. Stable HKB11 clones secreting high levels of a tissue factor (TF; 40-50 pg/c/d) and B-domain deleted recombinant factor VIII (BDDrFVIII; 5-10 microU/c/d) were derived under serum-free conditions. The specific productivity for these two proteins from the HKB11 cells was 10-fold greater than those from CHO cells derived under the similar conditions. In conclusion, we have demonstrated that the HKB11 cell line is well-suited for transient and long-term production of recombinant proteins.

[Cytotoxic effect of Acanthamoeba trophozoite on HeLa cells].

OBJECTIVE: To investigate the cytotoxic effect(CTE) on human cervix cancer HeLa cells induced by five strains of pathogenic free-living Acanthamoeba. METHODS: The cytotoxic effect of five isolates of Acanthamoeba on HeLa cells was investigated by light microscopy and MTT method. RESULTS: The photomicrographs of HeLa cells showed a sequence of cardinal morphological features of apoptosis when HeLa cells were exposed to Acanthamoeba in a time-dependent manner at a ratio of 1:1 for 12 h. MTT method showed more than 50% of tumor cells underwent cytolysis following exposure to A. lugdunensis trophozoites, and only 18% of cells treated with A. polyphaga underwent CTE. The CTE produced by A. lugdunensis and A. quina trophozoites was more rapid than the others, beginning as early as 6 h after coincubation and resulting in cytolysis by 72 h. CONCLUSION: These five strains of Acanthamoeba exhibit cytotoxic effects of varying degrees on HeLa cells, inducing apoptosis.

Function of pref-1 as an inhibitor of adipocyte differentiation.

During conversion of preadipocytes to adipocytes, growth arrest and subsequent activation of adipocyte genes by the transcription factors, C/EBPalpha and PPARgamma, lead to adipogenesis. During differentiation, these cells not only start expressing those genes necessary for adipocyte function, but also undergo changes in morphology to become rounded lipid filled adipocytes. Various factors in cell-cell communication or cell-matrix interaction may govern whether preadipocytes are kept in an undifferentiated state or undergo differentiation. In an attempt to identify molecules that play critical roles in the conversion of preadipocytes to adipocytes, we cloned by differential screening several regulatory molecules, including pref-1. Pref-1 is an inhibitor of adipocyte differentiation and is synthesized as a plasma membrane protein containing 6 EGF-repeats in the extracellular domain. Pref-1 is highly expressed in 3T3-L1 preadipocytes, but is not detectable in mature fat cells. Dexamethasone, a component of standard differentiation agents, inhibits pref-1 transcription and thereby promotes adipogenesis. Downregulation of pref-1 is required for adipose conversion and constitutive expression of pref-1 inhibits adipogenesis. Conversely, decreasing pref-1 levels by antisense pref-1 transfection greatly enhances adipogenesis. The ectodomain of pref-1 is cleaved to generate a biologically active 50kDa soluble form. There are four major forms of membrane pref-1 resulting from alternate splicing. Two of these forms which have a deletion that includes the putative processing site proximal to the membrane do not produce a biologically active soluble form. This indicates that alternate splicing may determine the range of action, juxtacrine or paracrine, of pref-1.

The ABA-1 allergen of Ascaris lumbricoides: sequence polymorphism, stage and tissue-specific expression, lipid binding function, and protein biophysical properties.

The ABA-1 protein of Ascaris lumbricoides (of humans) and Ascaris suum (of pigs) is abundant in the pseudocoelomic fluid of the parasites and also appears to be released by the tissue-parasitic larvae and the adult stages. The genes encoding the polyprotein precursor of ABA-1 (aba-1) were found to be arranged similarly in the two taxa, comprising tandemly repeating units encoding a large polyprotein which is cleaved to yield polypeptides of approximately 15 kDa which fall into 2 distinct classes, types A and B. The polyprotein possibly comprises only 10 units. The aba-1 gene of A. lumbricoides is polymorphic, and the majority of substitutions observed occur in or near predicted loop regions in the encoded proteins. mRNA for ABA-1 is present in infective larvae within the egg, and in all parasitic stages, but was not detectable in unembryonated eggs. ABA-1 mRNA was confined to the gut of adult parasites, and not in body wall or reproductive tissues. Recombinant protein representing a single A-type unit for the A. lumbricoides aba-1 gene was produced and found to bind retinol (Vitamin A) and a range of fatty acids, including the pharmacologically active lipids lysophosphatidic acid, lysoplatelet activating factor, and there was also evidence of binding to leukotrienes. It failed to bind to any of the anthelmintics screened. Differential Scanning Calorimetry showed that the recombinant protein was highly stable, and unfolded in a single transition at 90.4 degrees C. Analysis of the transition indicated that the protein occurs as a dimer and that the dimer dissociates simultaneously with the unfolding of the monomer units.

Secretion of a novel class of iFABPs in nematodes: coordinate use of the Ascaris/Caenorhabditis model systems.

A novel fatty acid binding protein, As-p18, is secreted into both the perivitelline and perienteric fluids of the parasitic nematode, Ascaris suum, and at least eight potential homologues of As-p18 have been identified in the Caenorhabditis elegans genome. The products of the three most closely related homologues are fatty acid binding proteins (LBP-1, LBP-2 and LBP-3) which contain putative secretory signals. Phylogenetic analysis revealed that these secreted fatty acid binding proteins comprise a distinct gene class within the fatty acid binding protein family and are possibly unique to nematodes. To examine the potential sites of As-p18 secretion, the expression of the putative promoters of the C. elegans homologues was examined with GFP reporter constructs. The developmental expression of lbp-1 was identical to that of As-p18 and consistent with the secretion of LBP-1 from the hypodermis to the perivitelline fluid. The expression patterns of lbp-2 and lbp-3 were consistent with the secretion of LBP-2 and LBP-3 from muscle into the perienteric fluid later in development. These studies demonstrate that at least some perivitelline fluid proteins appear to be secreted from the hypodermis prior to the formation of the cuticle and, perhaps more importantly, that this coordinate C. elegans/A. suum approach may be potentially useful for examining a number of key physiological processes in parasitic nematodes.