A density functional theory for ecology across scales.

Ecology lacks a holistic approach that can model phenomena across temporal and spatial scales, largely because of the challenges in modelling systems with a large number of interacting constituents. This hampers our understanding of complex ecosystems and the impact that human interventions (e.g., deforestation, wildlife harvesting and climate change) have on them. Here we use density functional theory, a computational method for many-body problems in physics, to develop a computational framework for ecosystem modelling. Our methods accurately fit experimental and synthetic data of interacting multi-species communities across spatial scales and can project to unseen data. As the key concept we establish and validate a cost function that encodes the trade-offs between the various ecosystem components. We show how this single general modelling framework delivers predictions on par with established, but specialised, approaches for systems from predatory microbes to territorial flies to tropical tree communities. Our density functional framework thus provides a promising avenue for advancing our understanding of ecological systems.

Alluvial fans at Cala Gonone (Sardinia), a fast developing touristic village: origins, hazards and potential risks.

The study area of Cala Gonone in NE Sardinia (Italy) consists of a wide terraced re-entrance/valley crowned inland by carbonate hills and, near the coast bounded laterally and partly floored by thin basaltic lava lying over carbonate bedrock. In this re-entrance, several inland alluvial fans (500 m length by 700 m wide) have developed, and a local ~ 30 m high, about 10 m wide (thick), 400 m long scarp body-remnant of semi-consolidated alluvial fan deposits is exposed along the coast. The fans experience depositional events mostly developed during the late Pleistocene. They although nowadays dormant may be reactivated by major rainstorms during strong climate changes. In these last few decades, the touristic village of Cala Gonone has been rapidly expanding over the mid to lower parts of two coalescing alluvial fans (Stadium and Gustui) and along the coastal marine scarp edge (Palmasera alluvial fan system). The village thus may become exposed to natural hazards if extreme climatic conditions may re-occur. Moreover, rock falls and the instability of the costal scarp due to wave erosion may add addition hazards for habitations built near the scarp crest and visitors to the frontal replenished beach. As commonly occurring elsewhere since antiquity, the risk perception of such events is low because of the centennial, millennial of longer recurrence. Such perception does not negate the hazards but a long event recurrence may be accepted as a reasonable risk for the human's activity. Nevertheless, serious consideration should be given to potential problems and plan and build for amelioration and defense. The evidence of what environmentally did and could still happen in the Cala Gonone and similar other area is in part clearly imprinted on the landscape: geology, geomorphology, and relative details in the stratigraphy and sedimentology of the deposits.

Heart urchins from the depths: Corparva lyrida gen. et sp. nov. (Palaeotropidae), and new records for the southwestern Atlantic Ocean / Erizos corazón de las profundidades: Corparva lyrida gen. et sp. nov. (Palaeotropidae) y nuevos registros para el Océano Atlántico sudoccidental

Introduction: Sea urchins in the order Spatangoida are the most diverse group of extant echinoids. Objective: Describe a new genus and species of Spatangoida from abyssal depths, and add new records for known species. Methods: Specimens were collected during several cruises at different areas of the southwestern Atlantic Ocean (SWAO), among 37-55° S latitude at depths ranging from 55 to 3 000 m. We present morphological and ultrastructure analyses. Results: Corparva lyrida gen. et sp. nov. (Palaeotropidae) is described from the Mar del Plata Canyon on the Argentine continental slope (2 950 m depth), the first record of this family from Argentina. Corparva gen. nov. differs in having an apical system semi-ethmolytic, and labrum reaching to rear part of second adjacent ambulacral plate. We also report the northernmost distribution and deepest record for Brisaster moseleyi (38° S latitude, 2 212 m depth), the northward extension of the distribution range of Tripylus excavatus (39° S latitude, 74 m depth), and the first record of Abatus philippii and Abatus agassizii at the Burdwood Bank/MPA Namuncurá. Conclusions: The present work brings novel and updated data about the diversity and distribution of spatangoids from the SWAO, including the description of C. lyrida gen. et sp. nov., and new records of species. This shows how much remains to be known about the diversity and distribution of heart urchins in the SWAO, especially from the deep-sea.

Introducción: Los erizos de mar del orden Spatangoida son el grupo más diverso de equinoideos recientes. Objetivo: Describir un nuevo género y una nueva especie de Spatangoida de profundidades abisales, y reportar nuevos registros para especies conocidas. Métodos: Los ejemplares fueron recolectados durante varias expediciones a diferentes áreas del Océano Atlántico sudoccidental (OAS), entre las latitudes 37-55° S y abarcando profundidades desde 55 a 3 000 metros. Presentamos análisis morfológicos y de ultraestructura. Resultados: Corparva lyrida gen. et sp. nov. (Palaeotropidae) fue descripta para el cañón submarino Mar del Plata en el talud continental de Argentina (2 950 m de profundidad), el primer registro de esta familia para Argentina. Corparva gen. nov. difiere en tener un sistema apical semi-etmolítico y labrum que llega a la parte posterior de la segunda placa ambulacral adyacente. También informamos la distribución más septentrional y el registro más profundo para Brisaster moseleyi (latitud 38° S, 2 212 m de profundidad), la extensión hacia el norte del rango de distribución de Tripylus excavatus (latitud 39° S, 74 m de profundidad) y el primer registro de Abatus philippii y Abatus agassizii en el Banco Burdwood/AMP Namuncurá. Conclusiones: El presente trabajo aporta datos novedosos y actualizados sobre la diversidad y distribución de erizos de mar espatangoideos del OAS, incluyendo la descripción de C. lyrida gen. et sp. nov., y nuevos registros para especies conocidas. Esto muestra cuánto queda por conocer sobre la diversidad y distribución de los erizos corazón en el OAS, especialmente de las profundidades marinas.

Impact of Vessel Suppressed-CT on Diagnostic Accuracy in Detection of Pulmonary Metastasis and Reading Time.

RATIONALE AND OBJECTIVES: To assess if vessel suppression (VS) improves nodule detection rate, interreader agreement, and reduces reading time in oncologic chest computed tomography (CT). MATERIAL AND METHODS: One-hundred consecutive oncologic patients (65 male; median age 60y) who underwent contrast-enhanced chest CT were retrospectively included. For all exams, additional VS series (ClearRead CT, Riverrain Technologies, Miamisburg) were reconstructed. Two groups of three radiologists each with matched experience were defined. Each group evaluated the SD-CT as well as VS-CT. Each reader marked the presence, size, and position of pulmonary nodules and documented reading time. In addition, for the VS-CT the presence of false positive nodules had to be stated. Cohen's Kappa (k) was used to calculate the interreader-agreement between groups. Reading time was compared using paired t test. RESULTS: Nodule detection rate was significantly higher in VS-CT compared to the SD-CT (+21%; p <0.001). Interreader-agreement was higher in the VS-CT (k = 0.431, moderate agreement) compared to SD-CT (k = 0.209, fair agreement). Almost all VS-CT series had false positive findings (97-99 out of 100). Average reading time was significantly shorter in the VS-CT compared to the SD-CT (154 ± 134vs. 194 ± 126; 21%, p<0.001). CONCLUSIONS: Vessel suppression increases nodule detection rate, improves interreader agreement, and reduces reading time in chest CT of oncologic patients. Due to false positive results a consensus reading with the SD-CT is essential.

Detection of hepatitis C virus in an exhumed body identified the origin of a nosocomial transmission that caused multiple fatal diseases.

BACKGROUND: Medico-legal conflicts arise when it is difficult to prove the cause of nosocomial infections. AIM: To report an outbreak of patient-to-patient transmission of hepatitis C virus (HCV) through the repeated use of a multi-dose saline flask during the rinsing of central venous catheters. METHODS: Blood samples were taken from each patient for the comparative analysis of their HCV RNA strains. No samples were available for one patient who died before the investigation started. Despite the known lability of HCV RNA, the body was exhumed four months after burial and postmortem samples were collected. HCV RNA was extracted successfully from liver and spleen samples. Genotyping of all the HCV strains was performed by sequence analysis of the 5'NC untranslated region, the E1 core conserved region and the E1/E2 hypervariable region. FINDINGS: Forensic investigators retraced the route used by two ward nurses, when saline catheter flushes were given to 14 patients with each nurse administering to seven patients. The comparative phylogenetic analysis of all case strains identified the deceased patient as the source of contamination to five patients. CONCLUSIONS: This study highlights the value of sequence analysis as a tool for solving medico-legal conflicts. The High Court of Justice found that a health worker's re-use of a contaminated needle resulted in the nosocomial transmission of HCV.

Urinary JCV-DNA testing during natalizumab treatment may increase accuracy of PML risk stratification.

The risk of progressive multifocal leukoencephalopathy (PML) in patients treated with natalizumab for multiple sclerosis (MS) is a serious concern. The presence of anti-JC virus antibodies is a risk factor for PML development, but 2.5 % of the patients result falsely-negative, while the prognostic relevance of testing JCV-DNA in biological fluids of treated patients is debated. Aim of this work was to evaluate the utility of testing JCV-DNA, together with anti-JCV antibodies, in biological samples of treated patients as a tool for PML risk stratification. 126 subjects from 5 MS Centers in Italy were included in the study. We performed a cross-sectional study in 63 patients testing JCV-DNA in blood, peripheral blood cells and urine. We longitudinally assessed the presence of JCV-DNA in a cohort of 33 subjects, one of which developed PML. We could test retrospectively serum samples from another PML case occurred during natalizumab therapy. Anti-JCV antibodies and urinary JCV-DNA were both tested in 73 patients. No changes in JCV-DNA status occurred during natalizumab treatment. The subject who developed PML in the longitudinal cohort had detectable JCV-DNA in urine at all time-points while serum or blood from both PML patients were always negative before the onset of disease and, in one case, after. Four subjects with JCV-DNA in urine and undetectable anti-JCV antibodies were retested for anti-JCV antibodies and three out of four resulted positive. In conclusion, testing JCV-DNA in urine is complementary to testing anti-JCV antibodies in identifying patients at risk of PML.

Fatty acids in formulae for term infants: compliance of present recommendations with the actual human milk fatty acid composition of geographically different populations.

AIM: Recommendations for formula fatty acids (FA) are largely based on the mature human milk FA composition. This study aimed to investigate whether current recommendations for formula FA for term infants comply with the actual breast-milk FA composition of geographically distinct populations and to provide more realistic grounds for future recommendations. METHODS: 455 mature breast-milk samples were collected in different countries over 25 y. Recommendations of different organizations were projected on their FA data. FA interrelationships were calculated with Spearman's rank tests. FA compositions of 30 formulae were compared with those of breast milk. RESULTS: Many samples from non-Western communities did not meet the recommendations for formula 12:0, 14:0 and 18:2omega6, since these are mainly based on breast milk of mothers living in Western countries. Recommendations for 18:3omega3, 18:2omega6/18:3omega3, 20:4omega6 and 22:6omega3 were not met by many milk samples, which may point to the poorly developed recommendations for long-chain polyunsaturated FA. Most of the investigated breast-milk FA (12:0, 14:0, 16:0, 18:0, 18:3omega3, 22:6omega3, 18:2omega6, 20:4omega6, 18:1omega9) were either positively or negatively interrelated. Many formulae had FA compositions that were not consistent with the physiological interrelationships of FA in breast milk. CONCLUSION: Future recommendations, if based on human milk, should derive from its FA balance, as indicated by the FA interrelationships. A "humanized" formula FA composition would in this sense be any composition that cannot be distinguished from that of breast milk by techniques such as principal component analysis.

Assessment of essential fatty acid and omega3-fatty acid status by measurement of erythrocyte 20:3omega9 (Mead acid), 22:5omega6/20:4omega6 and 22:5omega6/22:6omega3.

BACKGROUND: Early suspicion of essential fatty acid deficiency (EFAD) or omega3-deficiency may rather focus on polyunsaturated fatty acid (PUFA) or long-chain PUFA (LCP) analyses than clinical symptoms. We determined cut-off values for biochemical EFAD, omega3-and omega3/22:6omega3 [docosahexaenoic acid (DHA)]-deficiency by measurement of erythrocyte 20:3omega9 (Mead acid), 22:5omega6/20:4omega6 and 22:5omega6/22:6omega3, respectively. METHODS: Cut-off values, based on 97.5 percentiles, derived from an apparently healthy omnivorous group (six Dominica breast-fed newborns, 32 breast-fed and 27 formula+LCP-fed Dutch low-birth-weight infants, 31 Jerusalem infants, 33 Dutch 3.5-year-old infants, 69 omnivorous Dutch adults and seven Dominica mothers) and an apparently healthy group with low dietary LCP intake (81 formula-fed Dutch low-birth-weight infants, 12 Dutch vegans). Cut-off values were evaluated by their application in an EFAD suspected group of 108, mostly malnourished, Pakistani children, three pediatric patients with chronic fat-malabsorption (abetal-ipoproteinemia, congenital jejunal and biliary atresia) and one patient with a peroxisomal beta-oxidation disorder. RESULTS: Erythrocyte 20:3omega9, 22:5omega6/20:4omega6 and 22:5omega6/22:6omega3 proved age-dependent up to 0.2 years. Cut-off values for ages above 0.2 years were: 0.46mol% 20:3omega9 for EFAD, 0.068mol/mol 22:5omega6/20:4omega6 for omega3-deficiency, 0.22mol/mol 22:5omega6/22:6omega3 for omega3/DHA-marginality and 0.48mol/mol 22:5omega6/22:6omega3 for omega3/DHA-deficiency. Use of RBC 20:3omega9 and 22:5omega6/20:4omega6 cut-off values identified 20.4% of the Pakistani subjects as EFAD+omega3-deficient, 12.9% as EFAD+omega3-sufficient, 38.9% as EFA-sufficient+omega3-deficient and 27.8% as EFA-sufficient+omega3-sufficient. The patient with the peroxisomal disorder was classified as EFA-sufficient, omega3-sufficient (based on RBC 22:5omega6/20:4omega6) and omega3/DHA-deficient (based on RBC 22:5omega6/22:6omega3). The three other pediatric patients were classified as EFAD, omega3-deficient and omega3/DHA-deficient. CONCLUSION: Use of the combination of the present cut-off values for EFA, omega3 and omega3/DHA status assessment, as based on 97.5 percentiles, may serve for PUFA supplement intervention until better concepts have emerged.

Estimated biological variation of the mature human milk fatty acid composition.

We estimated the biological variation (CV(biol)) of 28 fatty acids (FA) in 465 mature human milk samples from The Netherlands, Caribbean, Jerusalem, Tanzania and Pakistan, by using data from the observed variation (CV(obs)) and analytical variation (CV(anal)). CV(biol) of the various regions was remarkably similar. The average CV(biol) of 455 samples, Pakistan excluded, ranged from 12.7% for 16:0 and 18.9% for 18:1 omega 9 to 68% for 22:6 omega 3 and about 100% for 20:5 omega 3. Those of 20:4 omega 6, 18:2 omega 6 and 18:3 omega 3 were 28.0, 33.0 and 37.3%, respectively. Because of the large CV(biol) and the many dietary changes in recent history, it seems impossible to consider the present human milk FA composition as the 'gold standard' for infant formula. Optimal human milk FA composition should rather derive from populations that consume traditional diets or from the scientific data that show the function of the individual FAs in neonatal development.

Optical control of electrons during electron transfer.

The dynamics of electron transfer reactions in solution can be controlled with the use of a sequence of femtosecond laser pulses. In the charge transfer to solvent (CTTS) reaction of sodide (Na-) in tetrahydrofuran, an initial light pulse launched the CTTS reaction, ejecting an electron into either an immediate or a solvent-separated Na0:solvated electron contact pair. A second pulse was used to excite the electrons in the contact pairs, and a third pulse monitored the amount of Na- produced through the back electron transfer. Excitation of the electrons in immediate contact pairs shut off the back electron transfer, whereas excitation of the electrons in solvent-separated pairs both enhanced and hindered the back electron transfer.

Parameters underlying successful protection with live attenuated mutants in experimental shigellosis.

Because the use of live attenuated mutants of Shigella spp. represents a promising approach to protection against bacillary dysentery (M. E. Etherridge, A. T. M. Shamsul Hoque, and D. A. Sack, Lab. Anim. Sci. 46:61-66, 1996), it becomes essential to rationalize this approach in animal models in order to optimize attenuation of virulence in the vaccine candidates, as well as their route and mode of administration, and to define the correlates of protection. In this study, we have compared three strains of Shigella flexneri 5--the wild-type M90T, an aroC mutant, and a double purE aroC mutant--for their pathogenicity, immunogenicity, and protective capacity. Protection against keratoconjunctivitis, induced by wild-type M90T, was used as the protection read out in guinea pigs that were inoculated either intranasally or intragastrically. Following intranasal immunization, the aroC mutant elicited weak nasal tissue destruction compared to M90T and achieved protection correlated with high levels of local anti-lipopolysaccharide immunoglobulin A (IgA), whereas the purE aroC double mutant, which also elicited weak tissue destruction, was not protective and elicited a low IgA response. Conversely, following intragastric immunization, only the M90T purE aroC double mutant elicited protection compared to both the aroC mutant and the wild-type strain. This mutant caused mild inflammatory destruction, particularly at the level of Peyer's patches, but it persisted much longer within the tissues. This could represent an essential parameter of the protective response that, in this case, did not clearly correlate with high anti-lipopolysaccharide IgA titers.

Short-term supplementation of low-dose gamma-linolenic acid (GLA), alpha-linolenic acid (ALA), or GLA plus ALA does not augment LCP omega 3 status of Dutch vegans to an appreciable extent.

Vegans do not consume meat and fish and have therefore low intakes of long chain polyunsaturated fatty acids (LCP). They may consequently have little negative feedback inhibition from dietary LCP on conversion of alpha -linolenic acid (ALA) to the LCP omega 3 eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids. We investigated whether supplementation of nine apparently healthy vegans with 2.01 g ALA (4 ml linseed oil), 1.17 g gamma-linolenic acid (GLA) (6 ml borage oil) or their combination increases the LCP omega 3 contents of erythrocytes (RBC) and platelets (PLT), and of plasma phospholipids (PL), cholesterol esters (CE) and triglycerides (TG). The supplements changed the dietary LA/ALA ratio (in g/g) from about 13.7 (baseline) to 6.8 (linseed oil), 14.3 (borage oil) and 6.4 (linseed + borage oil), respectively. ALA or GLA given as single supplements did not increase LCP omega 3 status, but their combination augmented LCP omega 3 (in CE) and EPA (in fasting TG) to a statistically significant, but nevertheless negligible, extent. We conclude that negative feedback inhibition by dietary LCP, if any, does not play an important role in the inability to augment notably DHA status by dietary ALA. The reach of a DHA plateau already at low dietary ALA intakes suggests that dietary DHA causes a non-functional DHA surplus, or is, alternatively, important for maintaining DHA status at a functionally relevant level.

The Lotus japonicus LjSym4 gene is required for the successful symbiotic infection of root epidermal cells.

The role of the Lotus japonicus LjSym4 gene during the symbiotic interaction with Mesorhizobium loti and arbuscular mycorrhizal (AM) fungi was analyzed with two mutant alleles conferring phenotypes of different strength. Ljsym4-1 and Ljsym4-2 mutants do not form nodules with M. loti. Normal root hair curling and infection threads are not observed, while a nodC-dependent deformation of root hair tips indicates that nodulation factors are still perceived by Ljsym4 mutants. Fungal infection attempts on the mutants generally abort within the epidermis, but Ljsym4-1 mutants allow rare, successful, infection events, leading to delayed arbuscule formation. On roots of mutants homozygous for the Ljsym4-2 allele, arbuscule formation was never observed upon inoculation with either of the two AM fungi, Glomus intraradices or Gigaspora margarita. The strategy of epidermal penetration by G. margarita was identical for Ljsym4-2 mutants and the parental line, with appressoria, hyphae growing between two epidermal cells, penetration of epidermal cells through their anticlinal wall. These observations define a novel, genetically controlled step in AM colonization. Although rhizobia penetrate the tip of root hairs and AM fungi access an entry site near the base of epidermal cells, the LjSym4 gene is necessary for the appropriate response of this cell type to both microsymbionts. We propose that LjSym4 is required for the initiation or coordinated expression of the host plant cell's accommodation program, allowing the passage of both microsymbionts through the epidermis layer.

Neuronal glycolytic pathway impairment induced by HIV envelope glycoprotein gp120.

Neurological impairment is a common feature of Acquired Immunodeficiency Syndrome (AIDS); functional alterations have been reported both in central and peripheral nervous system and the Human Immunodeficiency Virus (HIV) envelope glycoprotein gp120 has been proposed as a neurotoxin acting through a calcium-dependent mechanism. On the other hand it has been reported that gp120 treatment also induce about a 20% decrease in the cerebral glucose utilization and in the cellular ATP levels. The reported observations were performed on experimental system where also non-neuronal cells where present; in order to evaluate whether a direct interaction between HIV proteins and neuronal cells takes place, we used a neuroblastoma cultures where only neuronal cells are present. We analysed the effects of gp120 on the N18TG2 neuroblastoma clone. Treatments were performed both on growing and confluent cultures. Short time treatment with gp120 of confluent cultures causes a 25% reduction in the level of neuron-specific enolase, resulting in a similar decrease of oxygen consumption. Long time exposure of growing cells also causes a reduction in cell survival. Furthermore, using a membrane-specific fluorescent probe we observed that gp120 produces an increase of membrane trafficking. These observations suggest a direct interaction between the viral envelope protein and neuronal cells, which results in an alteration of glycolytic metabolism. This alteration may be related to the neurologic impairments observed in AIDS patients.

Level of human immunodeficiency virus DNA in peripheral blood mononuclear cells correlates with efficacy of antiretroviral therapy.

A novel colorimetric assay was developed and validated for accurate quantitation of human immunodeficiency virus (HIV) DNA in peripheral blood mononuclear cells (PBMCs). We tested 318 sequential samples from 56 subjects, 53 of whom were undergoing dual or triple therapy. Patients were considered responders when viremia levels were below 5, 000 HIV RNA copies/ml. The mean DNA copy numbers for untreated and responder subjects were similar (72 and 75, respectively), while it was 4.54-fold higher for nonresponders (339). This report provides strong evidence that HIV DNA levels in PBMCs correlate with therapeutic efficacy and suggests that DNA quantitation is a useful tool to monitor the decay of the HIV reservoir toward disease remission, especially when viremia is undetectable.

ArchAIDS connects a clinical department and a virological laboratory to automatize the follow up of AIDS patients.

A communication system for the automation of the follow up of AIDS patients set up by DIST at the Molecular Virology Unit in the Advanced Biotechnology Centre of Genova and at the Department of Internal Medicine of the Medical School of Genova is presented. This system includes a distributed database to store both clinical and virological data and a set of procedures to transfer patient data with a complete respect of requirements about completeness and privacy.

Human Immunodeficiency Virus Localization in Human Papillomavirus-Related, High-Grade Squamous Intraepithelial Lesions of the Cervix in Women with HIV Infection: Microdissection and Molecular Analysis on Formalin-Fixed and Paraffin-Embedded Specimens.

OBJECTIVES: To evaluate a possible mechanism of human immunodeficiency virus (HIV) and human papillomavirus (HPV) interaction, we have identified the cervical tissue compartments that harbor HIV. MATERIALS AND METHODS: We studied 39 paraffin-embedded, cervical conization specimens with high-grade cervical intraepithelial neoplasia (CIN3) occurring in HIV-infected women. From selected intraepithelial HPV-positive nonulcerated specimens (confirmed by in situ hybridization), we obtained serial 4- to 5-µm-thick sections that were stained with hematoxylin and eosin, anti-S100 protein, and anti-CD4. The presence of intramucosal Langerhans' cells or dendritic cells or CD4-positive cells was recorded. Three consecutive, nonmicrodissected, full-thickness sections of the same specimens were used for polymerase chain reaction (PCR) analysis (group A). Three other uncovered, consecutive sections from the same blocks were examined with an inverted microscope, and full-thickness specimens of mucosa were dissected from the underlying cervical stroma, were gently removed, and were used for PCR (group B). The quality of DNA was checked by HLA-DQa amplification; then a nested PCR for HIV proviral DNA was performed. RESULTS: Of group A, 5 of 39 cases (12.8%) were positive, whereas HIV was not detected in the microdissected sections of group B, with or without intraepithelial Langerhans' or CD4 cells. CONCLUSIONS: HIV does not affect cervical epithelium. The absence of infected Langerhans' or dendritic cells (or both) indicates a migration to the proximal lymph nodes of the in ….

A novel and innovative quantitative kinetic software for virological colorimetric assays.

This study addresses the limited range of quantification with colorimetric assays (ELISA) starting from the analysis of color production in a reference external curve. An automatic ELISA management software, designated Quanti-Kin Detection System (QKDS) is described, which retains the sensitivity of the end-point reading and extends the dynamic range up to five logarithms with mathematical interpretation of color production. The QKDS software is a generic system suitable for different types of ELISA with substrate incubation at room temperature, does not require dedicated instruments, performs accurate quantification (including assay quality control) and has a user friendly interface. Specific applications were developed for three types of analytes: antibodies, viral antigens and nucleic acids. Data are presented on three representative QKDS applications to HIV antibodies, p24 antigen and proviral DNA kits. The precision of quantification is strictly correlated with the precision of the kit; however, for almost all samples with known analyte amount, the error percentage was below 10%, only for two cases in quantification of HIV proviral DNA the error percentage was around 25%. The necessity for a wide quantification range has been demonstrated by measuring clinical samples, which showed a distribution in all possible quantification ranges for all kits.

Effects of erythropoietin administration on blood pressure and urinary albumin excretion in rats.

The effects of recombinant human erythropoietin (rHuEPO) administration on blood pressure and urinary albumin excretion were studied in normotensive Wistar-Kyoto rats (WKY), in spontaneously hypertensive rats (SHR), and in SHR rats treated with an angiotensin converting enzyme inhibitor (SHR-ACEi). Rats were housed in metabolic cages and treated with rHuEPO (150 U/kg body weight [bw] three times a week) for 6 weeks. Control animals received the vehicle only (0.25 mL of physiological saline). An angiotensin converting enzyme inhibitor was administered in the drinking water for 6 weeks (spirapril 5 mg/kg bw). Systolic blood pressure (SBP), and 24 h urinary albumin excretion (UAE) were measured once a week. No significant differences in SBP were observed between rHuEPO and vehicle-treated normotensive animals at the end of the treatment (171.9 +/- 4.9 v 172.1 +/- 5.6 mm Hg, respectively). After 6 weeks, SBP was significantly higher in SHR and SHR-ACEi groups treated with rHuEPO than in control groups (239.8 +/- 7.3 and 243.0 +/- 7.3 mm Hg v 218.1 +/- 6.0 and 187.9 +/- 4.6 mm Hg, respectively); UAE was significantly higher in groups treated with rHuEPO than in control groups (WKY: 265.9 +/- 19.5 v 127.0 +/- 12.3 microg/100 g bw, SHR: 1668.4 +/- 564.6 v 234.8 +/- 22.9 microg/100 g bw, and SHR-ACEi: 1522.7 +/- 448.3 v 143.0 +/- 18.9 microg/100 g bw, respectively). We concluded that erythropoietin treatment causes an increase in arterial pressure in SHR only, and an increase in UAE in both normotensive and hypertensive rats. The albuminuric effect was not entirely dependent on increased blood pressure. The treatment with an angiotensin converting enzyme inhibitor did not modify either the proteinuric or the pressor effects.

Overexpression, one-step purification and characterization of UDP-glucose dehydrogenase and UDP-N-acetylglucosamine pyrophosphorylase.

Two enzymes of the Leloir pathway, UDP-GlcNAc pyrophosphorylase and UDP-Glc dehydrogenase, which are involved in the synthesis of activated sugar nucleotides have been cloned, overexpressed in Escherichia coli, and purified to homogeneity in only one step by chelation-affinity chromatography. The gene KfaC of E. coli K5 was thus demonstrated to encode UDP-Glc DH. Some properties of the cloned enzymes, such as stability, pH dependence, and substrate kinetics, were studied in order to facilitate the use of these enzymes in carbohydrate synthesis, especially in the synthesis of hyaluronic acid.