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.

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.

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.

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.

Influence of docosahexaenoic acid on cisplatin resistance in a human small cell lung carcinoma cell line.

In a sensitive, human, small cell lung carcinoma cell line (GLC4) and a cisplatin (CP)-resistant subline (GLC4-CP), the effect of co-culturing with docosahexaenoic acid (DCHA) on CP cytotoxicity was studied. Cells were cultured for 4 days, with 32 microM of DCHA added on days 1 and 3. Incorporation of DCHA into the cellular phospholipids was demonstrated by fatty acid analysis. Supplementation with DCHA led to almost a threefold decrease of resistance in GLC4-CP and had no influence on CP cytotoxicity in GLC4. After culturing with DCHA, cellular platinum (Pt); total Pt bound to DNA; and Pt-GG, Pt-AG, G-Pt-G, and Pt-GMP adduct contents increased in both lines, whereas interstrand cross-link formation was elevated only in GLC4-CP. These experiments demonstrate that DCHA reduces CP resistance. Although an effect on cellular membranes resulting in an increased CP uptake apparently was present, this mechanism does not seem to be responsible for resistance modulation. Rather, an effect on nuclear, probably DNA-related, structures is likely and leads to an increased formation of interstrand cross-links in GLC4-CP.

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.

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.

Synthesis of the biologically active beta-subunit of human nerve growth factor in Escherichia coli.

The gene (NGFB) encoding the beta subunit of mature human nerve growth factor (hNGFB) was subcloned into the pJLA503 expression vector under the control of bacteriophage promoters PR and PL, and expressed in Escherichia coli. The recombinant protein represented approximately 3% of the total cellular protein. Biologically active hNGFB was solubilized (0.2% total NGFB) and purified by cation-exchange chromatography and it yielded two bands on polyacrylamide-gel electrophoresis under nonreducing conditions, corresponding to the monomeric (14 kDa) and homodimeric (26.5 kDa) forms of the molecule. Both hNGFB forms were immunopositive on Western blots with rabbit anti-NGFB antibodies; however, following additional purification, only the species corresponding to the hNGFB homodimer was biologically active on cultured chicken dorsal root ganglion neurons. These results demonstrate the feasibility of synthesizing the biologically active form of hNGFB in E. coli.

A carbohydrate-rich diet not only leads to incorporation of medium-chain fatty acids (6:0-14:0) in milk triglycerides but also in each milk-phospholipid subclass.

We isolated phospholipid (PL) subclasses from milk of women in Dominica and Belize. Fatty acid (FA) compositions of PLs and total lipids were determined. In the total-lipid fraction Dominican milk showed higher relative amounts of medium-chain saturated fatty acids (MC-SAFAs; 6:0-14:0) and 22:6n-3 and lower amounts of long-chain saturated fatty acids (LC-SAFAs) and monounsaturated fatty acids (MUFAs). There was a positive relationship between the MC-SAFA content in total lipids and total PLs. Incorporation of MC-SAFAs in PLs occurred at the expense of LC-SAFAs, MUFAs, polyunsaturated fatty acids (PUFAs), and long-chain PUFAs with greater than or equal to 20 carbon atoms (LC-PUFAs greater than or equal to C20). Previous studies from Western countries revealed low amounts of MCSAFAs and high amounts of PUFAs and LC-PUFAs greater than or equal to C20 in milk PLs. Our data show that carbohydrate-rich diets give rise to incorporation of MC-SAFAs in PLs at the expense of PUFAs and LC-PUFAs greater than or equal to C20. The data are discussed in relation to the presumed origin of fat-globule membrane phospholipids.

Fat absorption in neonates: comparison of long-chain-fatty-acid and triglyceride compositions of formula, feces, and blood.

We studied malabsorption of fat in neonates who were fed either a lard-modified formula (n = 22, gestational age, 33.6 +/- 3.9 wk) or an unmodified formula (n = 14, gestational age, 34.1 +/- 3.7 wk). In both groups fecal lipid consisted almost completely (greater than 90%) of free fatty acids, whose composition was highly correlated with the corresponding formula's fatty acid composition [r = 0.96 (lard modified) and r = 0.99 (standard)]. Both groups had similar relative amounts and compositions of fecal cholesterol esters and triglycerides. Plasma and, to a lesser extent, erythrocyte membrane long-chain-fatty-acid compositions correlated with those of the corresponding formulas. We suggest that the extensive intestinal hydrolysis and limited absorption of dietary lipids is, at least partly, due to lipolysis in the colon. Appearance of triglycerides in the colon may be due to a rapid small-intestinal passage in relation to small-intestinal lipolysis.

Milk of patients with tightly controlled insulin-dependent diabetes mellitus has normal macronutrient and fatty acid composition.

The composition of macro- and micronutrients in milk from six patients with tightly controlled insulin-dependent diabetes mellitus [median glycosylated hemoglobin concentrations at parturition of 5.2% (range 4.9-5.3%, reference range 4.9-6.6%) and 6 wk thereafter of 6.1% (range 5.0-6.3%, reference range 5.0-6.4%) was compared with that from five control subjects. Milk samples were collected halfway through a single breast-feeding at days 3-5 (colostrum); 7, 9, and 10 (transitional milk); and 12, 15, 17, 21, 25, 29, and 35 (mature milk). We found no abnormalities in macronutrient (triglycerides, lactose, and protein), cholesterol, glucose, and myoinositol concentrations or fatty acid composition. Two of three longitudinally studied patients showed rather constant ratios between glucose concentrations in milk and capillary blood. The present data suggest that tight control corrects a multitude of milk abnormalities associated with moderate and poor control.

HLA-DR Schwann cell reactivity in peripheral neuropathies of different origins.

HLA-DR antigens have been found on Schwann cells in peripheral neuropathies of different origins but not in normal control cases. Class II antigen reactivity was more intense in chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) and hereditary motor and sensory neuropathy type 1 (HMSN), but was also observed in toxic or metabolic neuropathies. The expression of HLA-DR antigen on Schwann cells does not appear to be related to the inflammatory or autoimmune origin of the disease.

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.

Rapid changes in serum, plasma and erythrocyte lipid compositions, and serum transaminase levels during continuous enteral hyperalimentation by carbohydrates alone.

Twelve normal men received twice their estimated basal energy requirement by a carbohydrate solution via a nasogastric catheter during 48 hours, followed by a seven-hour fast. Subsequently, in nine of them 0.5 mg epinephrine was given subcutaneously under ongoing fasting. During hyperalimentation, serum triglycerides, phospholipids, total and free cholesterol, phospholipids/free cholesterol ratio, and plasma free fatty acids decreased, whereas the percentage of free cholesterol increased. During fasting and subsequent epinephrine administration triglycerides and free fatty acids rose without reaching basal levels. Plasma and red blood cell (RBC) fatty acid composition already changed from two hours after the start of the feeding. Most markedly, a steady decrease in RBC 18:2c, omega 6, amounting to more than 17% of the basal value at the end of the observation period was found. Neither in plasma, nor in RBC a concomitant appearance of 20:3c, omega 9 was seen. In RBC, the relative amounts of the saturated fatty acids increased, whereas those of monounsaturated and polyunsaturated fatty acids decreased. RBC content of total fatty acids decreased and that of cholesterol increased. The ratios 16:1c, omega 7/16:0 and 18:3c, omega 6/18:2c, omega 6 in plasma, and 20:3c, omega 6/18:2c, omega 6 in plasma and RBC increased, whereas those of 18:1c, omega 7/16:1c, omega 7 and 20:3c, omega 6/18:3c, omega 6 in plasma decreased. After 48 hours feeding serum glutamic pyruvic transaminase and glutamic oxaloacetic transaminase levels were moderately increased and rose further during fasting. Thus, continuous enteral hyperalimentation by carbohydrates alone rapidly induces profound changes in serum-, plasma-, and RBC lipid compositions and serum parameters of hepatic function.(ABSTRACT TRUNCATED AT 250 WORDS)

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.

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.

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.

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.

GFAP expression of human Schwann cells in tissue culture.

We have studied the expression of the intermediate filament (IF) proteins, vimentin and glial fibrillary acidic protein (GFAP), in cultured human Schwann cells (SC) from patients with different neuropathies and normal control cases. SC cultures from sural nerve biopsies of 8 subjects with axonal neuropathies, 8 with demyelinating neuropathies and 3 normal controls were included in this study and processed with double immunofluorescence technique, using anti-vimentin and anti-GFAP antibodies, during the 2nd, 4th and 6th week of culture. Five cultures incubated with anti-GFAP antibodies were also processed for immunoelectron microscopy. Specificity tests of the used antibodies were performed. We have found that: (1) cultured human SC constantly express vimentin; (2) SC from normal controls are GFAP-negative in the first period of culture; (3) SC from pathologic nerves can contain GFAP-immunoreactive IF and the percentage of GFAP-positive SC is higher in axonal than in demyelinating neuropathies; (4) during the permanence in culture human SC from both normal and pathologic cases acquire the ability to synthesize GFAP. The obtained data suggest that the removal from axonal contact and the resulting loss of myelinating function induce a cytoskeletal cellular response in human SC characterized by the cytoplasmic accumulation of GFAP-immunoreactive IF.