Counteract of bone marrow of blotchy mice against the increases of plasma copper levels induced by high-fat diets in LDLR−/− mice

BackgroundBone marrow of blotchy mouse (blotchy marrow) reflects the function of transmembrane domain and relevant intramembrane sites of ATP7A in myeloid cells. By chronic infusion of angiotensin II, we previously found that blotchy marrow plays a minor role in regulating plasma copper. Moreover, the recipients of blotchy marrow presented a moderate reduction of plasma lipids and inflammatory mediator production. Little is known about whether these changes are a specific response to angiotensin II or reveal a more general role of ATP7A.Objective and designWe investigated if blotchy marrow reduces plasma lipids and inflammatory mediators induced by high-fat diets. To test this hypothesis, blotchy and control marrows were reconstituted to the recipient mice (irradiated male LDLR−/− mice), followed by high-fat-diet feeding for 4 months. At the end points, plasma metals (copper, zinc and iron), lipid profiling (cholesterol, triglyceride, phospholipids and lipoprotein) and six inflammatory mediators (lymphotacin, MCP3, MCP5, TIMP1, VEGF-A and IP-10) were measured. Parallel experiments were performed using male LDLR−/− mice fed either high-fat diets or chow diets for 4 months.ResultsIn addition to hyperlipidemia and low-grade inflammation, high-fat diets selectively increased plasma copper concentration compared to chow diets in LDLR−/− mice. After high-fat-diet feeding, the recipients with blotchy marrow showed a decrease in plasma copper (p < 0.01) and an increase in plasma iron (p < 0.05). The recipients with blotchy marrow also presented decreases in cholesterol (p < 0.01) and phospholipids (p < 0.05) in plasma. Surprisingly, plasma levels of MCP3 (p < 0.05), MCP5 (p < 0.05), TIMP1 (p < 0.01), VEGF-A (p < 0.01) and IP-10 (p < 0.01) were significantly increased in the recipients with blotchy marrow compared to controls; the increased levels of MCP3, MCP5 and TIMP1 were more than 50%.ConclusionOur studies showed that blotchy marrow counteracts the increased copper levels induced by high-fat diets, indicating that circulating myeloid cells can regulate blood copper levels via ATP7A. Moreover, transplantation of blotchy marrow followed by high-fat diets leads to a decrease in lipid profile and an increase in inflammatory mediator production. Overall, blotchy marrow mediates divergent responses to angiotensin II and high-fat diets in vivo.     

Viral safety of C1-inhibitor NF

We studied the efficacy of virus reduction by three process steps (polyethylene glycol 4000 (PEG) precipitation, pasteurization, and 15 nm virus filtration) in the manufacturing of C1-inhibitor NF. The potential prion removing capacity in this process was estimated based on data from the literature. Virus studies were performed using hepatitis A virus (HAV) and human immunodeficiency virus (HIV) as relevant viruses and bovine viral diarrhea virus (BVDV), canine parvovirus (CPV) and pseudorabies virus (PRV) as model viruses, respectively. In the PEG precipitation step, an average reduction in infectious titer of 4.5 log₁₀ was obtained for all five viruses tested. Pasteurization resulted in reduction of infectious virus of >6 log₁₀ for BVDV, HIV, and PRV; for HAV the reduction factor was limited to 2.8 log₁₀ and for CPV it was zero. Virus filtration (15 nm) reduced the infectious titer of all viruses by more than 4.5 log₁₀. The overall virus reducing capacity was >16 log₁₀ for the LE viruses. For the NLE viruses CPV and HAV, the overall virus reducing capacities were >8.7 and >10.5 log₁₀, respectively. Based on literature and theoretical assumptions, the prion reducing capacity of the C1-inhibitor NF process was estimated to be >9 log₁₀.     

Liquid chromatography–tandem mass spectrometry assay for the quantification of free and total sialic acid in human cerebrospinal fluid

BackgroundAnalysis of sialic acid (SA) metabolites in cerebrospinal fluid (CSF) is important for clinical diagnosis. In the present study, a high-performance liquid chromatography–tandem mass spectrometry (HPLC/MS/MS) method for free sialic acid (FSA) and total sialic acid (TSA) in human CSF was validated.MethodsThe method utilized a simple sample-preparation procedure of protein precipitation for FSA and acid hydrolysis for TSA. Negative electrospray ionisation was used to monitor the transitions m/z 308.2 → 87.0 (SA) and m/z 311.2 → 90.0 (¹³C₃-SA). Conjugated sialic acid (CSA) was calculated by subtracting FSA from TSA. We established reference intervals for FSA, TSA and CSA in CSF in 217 control subjects. The method has been applied to patients’ samples with known differences in SA metabolites like meningitis (n = 6), brain tumour (n = 2), leukaemia (n = 5), and Salla disease (n = 1).ResultsLimit of detection (LOD) was 0.54 μM for FSA and 0.45 μM for TSA. Intra- and inter-assay variation for FSA (21.8 μM) were 4.8% (n = 10) and 10.4% (n = 40) respectively. Intra- and inter-assay variation for TSA (35.6 μM) were 9.7% (n = 10) and 12.8% (n = 40) respectively. Tested patients showed values of TSA above established reference value.ConclusionThe validated method allows sensitive and specific measurement of SA metabolites in CSF and can be applied for clinical diagnoses.     

Influences of short-term pre-slaughter dietary manipulation in sheep and goats on pH and microbial loads of gastrointestinal tract

Sheep (BW = 39.9 kg, n = 16) and goats (BW = 32.8 kg, n = 16) were used in a completely randomized design to determine the effect of short-term pre-slaughter diet and feed deprivation (FD) time on pH and microbial loads in the gastrointestinal tract (GIT) contents. In a 2 × 2 × 2 factorial treatment arrangement, the main effects of species, diet, and FD time prior to slaughter and their interactions were studied. Animals were fed either a hay or concentrate diet for 4 d and then feed deprived for either 12 or 24-h prior to slaughter. The pH of rumen and colon contents as well as weight of GIT was measured. The contents of rumen and rectum were also sampled for microbial analysis. The GIT of sheep (1.82 kg) was heavier (P < 0.05) than that of goats (1.46 kg). The 12-h FD group (1.74 kg) had a higher (P < 0.05) GIT weight than the 24-h FD group (1.53 kg). Hay-fed animals had higher (P < 0.05) rumen (7.08 vs. 6.43) and colon pH values (7.02 vs. 6.56) than those of the concentrate-fed animals. The 24-h FD group (3.39 ± 0.272 log₁₀CFU/g) contained more (P < 0.05) Escherichia coli in the rumen than did the 12-h FD (2.17 ± 0.272 log₁₀CFU/g) group. The concentrate-fed animals (3.49 ± 0.289 log₁₀CFU/g) had higher (P < 0.05) coliform counts in the rumen than the hay-fed animals (2.43 ± 0.289 log₁₀CFU/g). The 24-h FD group (3.42 ± 0.289 log₁₀CFU/g) had a higher (P < 0.05) concentration of coliform than did the 12-h FD group (2.50 ± 0.289 log₁₀CFU/g). The 24-h FD group (3.31 ± 0.259 log₁₀CFU/g) also had higher (P < 0.05) Enterobacteriaceae counts in the rumen than did in the 12-h FD group (2.47 ± 0.259 log₁₀CFU/g). Goats (5.71 ± 0.158 log₁₀CFU/g) had lower (P < 0.05) total plate counts in the rumen compared to sheep (6.27 ± 0.158 log₁₀CFU/g). The concentrate-fed animals had higher (P < 0.05) E. coli (6.44 vs. 4.01 ± 0.468 log₁₀CFU/g), total coliform (6.74 vs. 4.16 ± 0.469 log₁₀CFU/g), Enterobacteriaceae (6.93 vs. 3.83 ± 0.651 log₁₀CFU/g), and total plate counts (7.79 vs. 7.28 ± 0.170 log₁₀CFU/g) in the rectum than the hay-fed animals. The results indicate that microbial loads in the GIT of small ruminants may be reduced by either feeding hay for 4 d or depriving feed for 12-h prior to slaughter.     

Involvement of the nitric oxide/CGMP/KATP pathway in antinociception induced by exercise in rats

AimsPhysical exercise is responsible for increasing the nociceptive threshold. The present study aimed to investigate the involvement of the nitric oxide/_CGMP/K_ATP pathway in antinociception induced by acute aerobic exercise (AAc) in rats.Main methodsWistar rats performed exercise in a rodent treadmill, according to an AAc protocol. The nociceptive threshold was measured by mechanical and thermal nociceptive tests (paw-withdrawal, tail-flick and face-flick). To investigate the involvement of the NO/_CGMP/K_ATP pathway the following nitric oxide synthase (NOS) unspecific and specific inhibitors were used: N-nitro-l-arginine (NOArg), Aminoguanidine, N⁵-(1-Iminoethyl)-l-ornithine dihydrocloride (L-NIO), N^ω-Propyl-l-arginine (L-NPA); guanylyl cyclase inhibitor, 1H-[1,2,4]oxidiazolo[4,3-a]quinoxalin-1-one (ODQ); and K_ATP channel blocker, Glybenclamide; all administered subcutaneously at a dose of 2 mg/kg 10 min before exercise started. Plasma and cerebrospinal fluid (CSF) nitrite levels were determined by spectrophotometry.Key findingsIn the paw-withdrawal, tail-flick and face-flick tests, the AAc protocol produced antinociception, which lasted for more than 15 min. This effect was significantly reversed (P < 0.05) by NOS specific and unspecific inhibitors, guanylyl cyclase inhibitor (ODQ) and K_ATP channel blocker (Glybenclamide). Acute exercise was also responsible for increasing nitrite levels in both plasma and cerebrospinal fluid.SignificanceTaken together, these results suggest that the NO/_CGMP/K_ATP pathway participates in antinociception induced by exercise.     

The plasma levels of CST and BCKDK in patients with sepsis

ObjectivesCST has been recently identified as a mediator of various beneficial effects in animal models of sepsis. At present, no data are available concerning the levels of CST in sepsis patients. In sepsis the plasma amino acid pattern is characterized by decreased branced chain amino acids (BCAAs). We investigated the levels of plasma CST or branched-chain α-ketoacid dehydrogenase kinase (BCKDK) and their relationship to component traits in patients with sepsis.Design and methodsWe studied 228 patients and divided them into two groups based on severity of infection. Blood samples were taken at study entry, and CST, BCKDK were measured.ResultsCST and BCKDK levels were significantly higher in patients with sepsis than in controls: the median plasma CST concentration was 103.1 ng/ml (range, <83.13–189.7 ng/ml) in patients with sepsis and 49.69 ng/ml (range, <19.38–100.8 ng/ml) in controls (p = 0.0022); the median plasma BCKDK concentration was 801.7 ng/ml in sepsis group and 745 ng/ml in controls (p = 0.0292). Additionally, there was correlation between the plasma concentrations of CST and BCKDK in sepsis patients (r² = 0.6357, p < 0.01).ConclusionsWe conclude that the plasma levels of CST in sepsis patients were higher than in controls, and there is a relationship between CST and BCKDK in sepsis patients. Future experimental and clinical studies are needed to evaluate CST as a novel prognostic tool in sepsis patients and its potential therapeutic use in sepsis.     

Angiostrongylus cantonensis eosinophilic meningitis: A clinical study of 42 consecutive cases in French Polynesia

IntroductionIn endemic areas, eosinophilic meningitis is mainly caused by Angiostrongylus cantonensis. We describe a series of this poorly-known condition.MethodsRetrospective cohort study (2000–2012) including all patients diagnosed with eosinophilic meningitis in French Polynesia.ResultsForty-two patients (males: 61.9%, age: 22 (IQR 17–32)) were diagnosed with a serologically proven (n = 13) or probable A. cantonensis meningitis, mostly during the dry season (66.6%) and following the consumption of or prolonged contact with an intermediate/paratenic host (64.3%). No differential diagnosis was found in probable cases, in whom serological tests were performed earlier (7.5 days (6.5–10)) compared to positive patients (7.5 (6.5–10) versus 11 (7–30) days, p = 0.02). The most commonly reported symptom was headache (92.8%). Fever (7.1%) and biological inflammatory syndrome (14.3%) were rare. Blood eosinophil count was 1200/mm³ (900–2548). Cerebrospinal fluid (CSF) analysis disclosed a protein level of 0.9 g/L (0.7–1.1), a CSF/plasma glucose ratio of 0.50 (0.40–0.55), and 500 leucocytes/mm³ (292–725; eosinophils: 42.0% (29.5–60); lymphocytes: 46.5% (32.5–59.0)). Thirteen cases (31.0%) were severe, with 11 focal neurological deficits. A delayed hospital referral (OR 1.13, p = 0.05) was associated with severity.ConclusionsA. cantonensis meningitis must be evocated in young patients with meningitic syndrome, severe headache, and CSF inflammation with predominance of eosinophils.     

Wide-spectrum profile of inflammatory mediators in the plasma and scales of patients with psoriatic disease

Psoriasis is a chronic recurrent inflammatory disorder of the skin. Clinical subtypes include psoriasis vulgaris (PV), psoriatic arthropathy, and erythrodermic psoriasis. Aim of this study was to analyse relevant inflammatory mediators in the plasma of patients with distinct subtypes of active psoriasis, and in the scales of mild-to-moderate PV patients, and correlation to disease severity. Compared to healthy controls (n = 10), patients affected by very severe forms of psoriasis (n = 30) were characterized by increased plasma levels of IL-4, IL-6, MCP-1, VEGF and in particular PDGFbb. Each group with severe psoriasis had distinct characteristic features of plasma cytokine profile. Mild-to-moderate PV patients (n = 35) showed higher levels of IL-4, IL-6, IL-10, and IL-13 when compared to healthy controls. No correlation was found between PV severity assessed by PASI (Psoriasis Area and Severity Index) and levels of these mediators. By contrast, disease severity correlated to scale levels of IP-10. For the first time, we found exaggerated circulating levels of the pro-angiogenic PDGFbb and VEGF in severe psoriasis. Evidence that the severity of skin symptoms correlated exclusively with scale levels of IP-10, but not with any up-regulated inflammatory mediator in plasma, suggests that distinct skin-independent processes contribute to the circulating cytokine profile in psoriasis.     

Plasma zinc in adults with cystic fibrosis: Correlations with clinical outcomes

BackgroundZinc status has been previously documented in cystic fibrosis (CF) infants, children and adolescents. However, despite the increasing life expectancy observed in CF populations, data regarding zinc status of CF adults are surprisingly lacking. The objectives of this study were to (1) characterize zinc status and (2) explore associations between zinc status and clinical outcomes of CF adult patients.MethodsA retrospective chart review was performed for patients who had their plasma zinc measured between 2009 and 2012. Data included demographics, clinical characteristics, biochemical parameters and co-morbid conditions.ResultsA total of 304 CF patients were included in the study. These patients displayed a good nutritional status (mean BMI ± SD: 22.7 ± 3.5) and moderate lung disease (mean FEV₁ ± SD: 66.3 ± 22.2). Low plasma zinc concentration (<9.2 μmol/L) was found in 68 out of 304 CF patients (22.4%). Compared to patients with normal zinc, those with low zinc had significantly lower forced vital capacity and forced expiratory volume in one second. 72% of CF adults with low zinc suffered from bone disease (vs 49% with normal zinc, p = 0.037) and 79% had impaired glycemic status (vs 58%, p = 0.016). Accordingly, negative correlations were found between plasma zinc and glucose (r = −0.139, p = 0.0001), HbA1c (r = −0.237, p = 0.0001) and fructosamine (r = −0.134, p = 0.034). In multiple linear regression, albumin and glycemic status were significant predictors of plasma zinc.ConclusionOur data indicated that nearly one quarter of CF adults with good nutritional status and moderate lung disease had low plasma zinc concentration and that low zinc status was associated with worse clinical outcomes.     

High levels of plasma selenium are associated with metabolic syndrome and elevated fasting plasma glucose in a Chinese population: A case-control study

BackgroundSelenium is important for human health and involved in various metabolic processes. Deficiency of selenium associates with increased risk for cancer and cardiovascular diseases. There has been an increase use of selenium supplements for the treatment of autoimmune thyroid conditions. However, the potential biological effects of selenium overload arouse the public concern. The aim of this study was to investigate the associations of plasma selenium concentrations of adults with metabolic syndrome (MS) in Chinese population.MethodsA matched case-control study including 204 metabolic syndrome patients and 204 healthy controls was conducted in 2012. The MS cases were defined according to the criteria of Chinese Diabetes Society (CDS). Healthy controls without abnormality of metabolic components were matched with cases in age, gender and region. Plasma concentrations of selenium were determined by graphite furnace atomic absorption spectrometry (GFAAS). Fasting plasma glucose (FPG), total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL), and low density lipoprotein cholesterol (LDL) were detected by automatic biochemical analyzer.ResultsThe median levels of plasma selenium in MS group were 146.3 (107.3–199.4) μg/L, which were significantly higher than that in the control group (127.4: 95.7–176.0) μg/L; Plasma levels of selenium were related to the risk of MS in dose-response manner. Risk of MS was significantly higher in subjects with plasma selenium in the highest tertile (T3: ≥176.0 μg/L) compared to those in the lowest tertile (T1: <95.7 μg/L) [odds ratio (OR) = 2.416 (95% CI: 1.289–4.526)]. The plasma levels of selenium were positively correlated with fasting plasma glucose (FPG) (r_s = 0.268, P < 0.001). Plasma selenium at the median (T2: 95.7–176.0 μg/L) or upper tertile (T3: ≥176.0 μg/L) was associated with increased risk of elevated FPG (defined by FPG ≥ 6.1 mmol/L) as compared with the lowest tertile (T1: ≤95.7 μg/L) [T2 vs. T1, OR = 3.487 (1.738–6.996); T3 vs. T1, OR = 6.245 (3.005–12.981)].ConclusionsHigher levels of plasma selenium might increase the risk of metabolic syndrome and elevated fasting plasma glucose. Selenium supplements should be used with prudence for CVD and cancer prevention.     

Multiple sclerosis: Identification and clinical evaluation of novel CSF biomarkers

Multiple sclerosis (MS) is a neuro-inflammatory and neurodegenerative disease that results in damage to myelin sheaths and axons in the central nervous system and which preferentially affects young adults. We performed a proteomics-based biomarker discovery study in which cerebrospinal fluid (CSF) from MS and control individuals was analyzed (n = 112). Ten candidate biomarkers were selected for evaluation by quantitative immunoassay using an independent cohort of MS and control subjects (n = 209). In relapsing–remitting MS (RRMS) patients there were significant increases in the CSF levels of alpha-1 antichymotrypsin (A1AC), alpha-1 macroglobulin (A2MG) and fibulin 1 as compared to control subjects. In secondary progressive MS (SPMS) four additional proteins (contactin 1, fetuin A, vitamin D binding protein and angiotensinogen (ANGT)) were increased as compared to control subjects. In particular, ANGT was increased 3-fold in SPMS, indicating a potential as biomarker of disease progression in MS. In PPMS, A1AC and A2MG exhibit significantly higher CSF levels than controls, with a trend of increase for ANGT. Classification models based on the biomarker panel could identify 70% of the RRMS and 80% of the SPMS patients correctly. Further evaluation was conducted in a pilot study of CSF from RRMS patients (n = 36), before and after treatment with natalizumab.     

Comparison of the peripheral mediator background of heat injury- and plantar incision-induced drop of the noxious heat threshold in the rat

AimsPreviously we described the drop of the noxious heat threshold in response to mild heat injury or plantar incision. While mild heat injury elicits an immediate and short-lasting thermal hyperalgesia, surgical incision leads to a delayed and sustained heat hyperalgesia. Only very few peripheral mediators of these phenomena have been identified. Therefore the present study aimed at comparing the peripheral mediator background of heat hyperalgesia evoked by mild heat injury or surgical incision.Main methodsHeat hyperalgesia was assessed by measuring the behavioural noxious heat threshold in conscious rats employing an increasing-temperature water bath.Key findingsThe heat threshold drop evoked by a mild heat injury and measured 10 min afterwards was reduced by intraplantarly applied HOE 140, a bradykinin B₂ receptor antagonist, NDGA, a non-selective lipoxygenase inhibitor, L-NOARG, a non-selective nitric oxide synthase inhibitor, TNP-ATP, a P2X purinoceptor antagonist and AMG9810, an antagonist of the transient receptor potential vanilloid type 1 (TRPV1) receptor. The heat threshold drop evoked by plantar incision and measured 18 h later was reduced by intraplantarly applied HOE 140, [des-Arg¹⁰]-HOE 140, a bradykinin B₁ receptor antagonist, L-NOARG, TNP-ATP and the TRPV1 receptor antagonist SB-366791.SignificanceOnly small differences have been revealed between the examined peripheral mediators of the acute heat hyperalgesia evoked by mild heat injury and the sustained increase in heat responsiveness induced by surgical incision. The B₂ and B₁ bradykinin receptor, P2X purinoceptors, TRPV1 receptor, nitric oxide synthase and lipoxygenase(s) are involved in at least one of these hyperalgesia models.     

Microbial community in conventional and extended aeration activated sludge plants in India

This study was conducted to determine the relationship between the ciliated protozoan population density and the effluent quality at two different modes of activated sludge plants (ASP) operating in India. A wide variety of ciliated protozoa (26 sp.) in higher density were identified at the conventional ASP, Haridwar, that delivered high quality effluent in terms of low biochemical oxygen demand (BOD = 15 mg/L), suspended solids (SS = 17 mg/L), turbidity (2.7 NTU), ammoniacal nitrogen (NH₃–N = 3 mg/L), chemical oxygen demand (COD = 37 mg/L), total coliforms (TC = log 5.2), fecal coliforms (FC = log 4.7) and fecal Streptococci (FS = log 3.7). Whereas, a few protozoan species (15 sp.) in lower density were reported in extended aeration plant (EAP) Delhi, that delivered turbid and lower quality effluent in terms of high BOD (23 mg/L), SS (80 mg/L), turbidity (12 NTU), NH₃–N (55 mg/L) and COD (68 mg/L). However, in spite of relatively poor effluent quality, lower concentration of TC (log 4.2), FC (log 3.9) and FS (log 3.2) was observed in EAP, Delhi. The constant presence of two filamentous bacterial species (Beggiatoa and Spirillum) in extended aeration process can be considered as the probable reason of high coliforms removal, since filamentous bacteria are capable of removing organic as well as microbial pollutants from wastewater.     

Toxicokinetics and toxicological effects of single oral dose of fumonisin B1 containing Fusarium verticillioides culture material in weaned piglets

Toxicokinetics and the toxicological effects of culture material containing fumonisin B₁ (FB₁) were studied in male weaned piglets by clinical, pathological, biochemical and sphingolipid analyses. The animals received a single oral dose of 5 mg FB₁/kg of body weight, obtained from Fusarium verticillioides culture material. FB₁ was detected by HPLC in plasma collected at 1-h intervals up to 6 h and at 12-h intervals up to 96 h. FB₁ eliminated in feces and urine was quantified over a 96-h period and in liver samples collected 96 h post-intoxication. Blood samples were obtained at the beginning and end of the experiment to determine serum enzyme activity, total bilirubin, cholesterol, sphinganine (Sa), sphingosine (So) and the Sa/So ratio. FB₁ was detected in plasma between 30 min and 36 h after administration. The highest concentration of FB₁ was observed after 2 h, with a mean concentration of 282 μg/ml. Only 0.93% of the total FB₁ was detected in urine between 75 min and 41 h after administration, the highest mean concentration (561 μg/ml) was observed during the interval after 8 at 24 h. Approximately 76.5% of FB₁ was detected in feces eliminated between 8 and 84 h after administration, with the highest levels observed between 8 and 24 h. Considering the biochemical parameters, a significant increase only occurred in cholesterol, alkaline phosphatase and aspartate aminotransferase activities. In plasma and urine, the highest Sa and Sa/So ratios were obtained at 12 and 48 h, respectively.     

Sewage treatment in a combined up-flow anaerobic sludge blanket (UASB)–down-flow hanging sponge (DHS) system

The performance of up-flow anaerobic sludge blanket (UASB) in combination with down-flow hanging sponge (DHS) system for sewage treatment at an average wastewater temperature of 15 °C has been investigated for 6 months. The results showed that a combined system operated at a total HRT of 10.7 h and total SRT of 88 days represents a cost effective sewage treatment process. The average COD_total and BOD_5 total concentrations measured in the final effluent of the total system (UASB + DHS) amounted to 43 and 3.0 mg/l, respectively, corresponding to the overall removal efficiency of 90% for COD_total and 98% for BOD_5 total. The total process provided a final effluent containing a low concentration of 12 mg/l for TSS.Eighty-six percent of ammonia was eliminated at space loading rate of 1.6 kg COD/m³ d and HRT of 2.7 h. The calculated nitrification rate of the DHS system according to the nitrate and nitrite production amounted to 0.18 kg/m³ d. The removal of F. coliform in the UASB reactor only amounted to 0.86 log₁₀. On the other hand, the F. coliform concentration dropped substantially, i.e. by 2.6 log₁₀ in the DHS system resulting only 2.7 × 10³/100 ml in the final effluent.The calculated average sludge production for UASB operated at an HRT of 8.0 h amounted to 30 g TSS/d, corresponding to sludge yield coefficient of 0.2 g TSS/g total COD removed, while it was indeed very low only 6.0 g TSS/d corresponding to sludge yield coefficient of 0.09 g TSS/g total COD removed, for DHS system.The DHS profile results revealed that in the first and second segment of DHS system, the COD_total, BOD_5 total and TSS was eliminated, followed by the oxidation of ammonia in the next segments.     

Modélisation des flux cérébraux par une analogie électrique : validation par vélocimétrie IRM

The cine Phase-Contrast Magnetic Resonance (PCMR) sequence is the only noninvasive technique for the study of cerebrospinal fluid (CSF) oscillations. It can provide CSF and blood flow measurements throughout the cardiac cycle. To study cerebral hydro-hemodynamic, models have been developed; nevertheless the majority of these models did not take into account the CSF oscillations. The objective of this study was to establish reference values for cerebral hydro-hemodynamic and propose a new electrical model of the brain dynamics.Material and methodsCSF and blood flows were measured in 19 control subjects by PCMR imaging. Dynamic flow images were analyzed on dedicated software to reconstruct the flow curves during the cardiac cycle. An electrical analogue was realized. The inputs of the model were fed by PCMR arterial and venous flows to simulate CSF oscillations. The simulated CSF oscillations were compared to the measured CSF oscillations to validate the model.ResultsThe key parameters of the CSF and blood flow curves were obtained, e.g. total cerebral blood flow was 688 ± 115 mL/min, ventricular CSF oscillatory volume was 0.05 ± 0.02 mL/cardiac cycle, and the subarachnoid CSF oscillatory volume was 0.55 ± 0.15 mL/cardiac cycle. A close agreement was found between measured and simulated cerebral CSF oscillations.ConclusionThis study established the main values characterizing cerebral hydrodynamics in a control population. It provided a better understanding of the mechanisms of intracranial volumes regulation during the cardiac cycle. Our results are now used in clinical practice and the model proposed is effective to study cerebral hydro-hemodynamic.     

Stability constants and multinuclear NMR measurements of phosphonic acid derivatives with aluminum in aqueous solutions

We are reporting the stability constants of the different complexes between phosphonoacetic acid (PAA), phosphonoformic acid (PFA), aminomethylphosphonic acid (AMPA), aminoethylphosphonic acid (AEPA) and methylenediphosphonic acid (MDP) with the aluminum metal ion in aqueous solutions. (In this study the term aluminum is reserved for the 3+ ion.) The affinity of the phosphonic acid derivatives to chelate aluminum has been tested by potentiometric titrations with I = 0.10 M KNO₃ at 25 ± 0.1 °C. The proposed aluminum–ligand complex structures have been confirmed by ³¹P NMR, ¹³C NMR, and ²⁷Al NMR experiments. Both PAA and PFA formed simple one to one complexes. The respective values for PAA are log β₁₁₁ = 13.57, log β₁₁₀ = 10.58, and log β₁₁₋₁ = 5.84. The respective values for PFA are log β₁₁₂ = 15.24, log β₁₁₁ = 13.11, and log β₁₁₀ = 6.88. In contrast to PAA and PFA, the major species formed with AMPA and AEPA consist of a series of dimeric complexes. The ³¹P NMR spectra of these complexes indicate that the amine groups do not co-ordinate to aluminum and remain protonated. In addition to these dimeric complexes, a monoprotonated monomer of Al–AMPA also has been detected. The ²⁷Al NMR experiments indicated that the aluminum is hexacoordinated in all the complexes in this study and the hydroxide ion did not remove aluminium from its complexes. Among the ligands studied, PAA and PFA were able to solubilize aluminum at physiological pH. A comparison between the acidities and the chelating properties of the ligands used is presented.     

Assay of Free Thyroxine and Free Triiodothyronine in Fine-Needle Aspiration of Thyroid Nodules: a Useful and Low-Cost Assessment

ObjectiveTo evaluate whether analysis of thyroid hormones in fine-needle aspiration (FNA) of thyroid nodules can provide information about the functional status and the nature of the nodules.MethodsWe studied 4 groups of patients: group 1, 17 patients with autonomous hyperfunctioning thyroid nodules; group 2, 52 patients with cold nonfunctioning thyroid nodules; group 3, 12 patients with malignant thyroid nodules; and group 4 (control group), 10 patients with nonthyroid nodular lesions (enlarged parathyroid glands or lymph nodes). The assay of thyroid hormones was performed in FNA after the washing of needles and, with patient consent, also in normal thyroid parenchyma.ResultsThe free thyroxine (FT₄) and free triiodothyronine (FT₃) values were remarkably high in group 1 (mean, 5.5 ± 0.53 ng/dL and 27.6 ± 3.1 pg/mL, respectively; P < 0.05 versus group 2 and group 4, the control group). The levels of FT₄ and FT₃ were very low in group 3 (< 0.2 ng/dL and < 1.0 pg/mL, respectively; P < 0.05 versus group 2). Thyroglobulin values in FNA specimens were much higher than the normal range in human serum, but no significant differences were found between the various groups. The control group had low levels of FT₄ and FT₃ (< 0.2 ng/dL and < 1.0 pg/mL, respectively) in conjunction with low levels of thyroglobulin, whereas parathyroid hormone levels were high in parathyroid nodules.ConclusionThese results show that assay of FT₄ and FT₃ in FNA can yield information about the functional status of thyroid nodules and, indirectly, about the nature of nodules. In this era of sophisticated new molecular markers in FNA cytology, this low-cost diagnostic method can be readily performed in every laboratory. (Endocr Pract. 2004;10:311-316)     

Simultaneous HPLC–MS–MS quantification of 8-iso-PGF2α and 8,12-iso-iPF2α in CSF and brain tissue samples with on-line cleanup

Quantitation of isoprostanes such as 8-iso-PGF_2α and 8,12-iso-iPF_2α-VI in biological fluids has been proposed as a reliable test of oxidant stress and inflammation in a variety of disorders. This paper presents a liquid chromatography method with tandem mass spectrometry detection for the simultaneous analysis of these two isoprostanes in human CSF and brain tissue samples. An API 5000 triple quadrupole instrument (AB Sciex, Foster City, CA, USA) with an APCI ion source was used in this study. Aliquots of CSF samples (0.25 mL) were treated with a methanol:zinc sulfate mixture followed by on-line cleanup on an extraction column (Validated-C₁₈) with 0.1% formic acid. The brain tissue samples were homogenized and lipids were extracted using Folch solution. Solid-phase extraction columns (C₁₈) were used for the purification of the brain isoprostane fraction. Chromatographic separation was achieved using an analytical column (Synergi C₁₈ HydroRP) with 0.1% formic acid in water and a mixture of methanol:acetonitrile under isocratic conditions. The mass spectrometer was operated in the MRM scan and negative ion mode. The quadrupoles were set to detect the molecular ions [M?H]^? and high mass fragments of isoprostanes: m/z 353 → 193 amu (8-iso-PGF_2α) and m/z 353 → 115 amu (8,12-iso-iPF_2α-VI) and their deuterated internal standards: m/z 357 → 197 amu (8-iso-PGF_2α-d₄) and m/z 364 → 115 amu (8,12-iso-iPF_2α-VI-d₁₁). The lower limit of quantification was 2.5 pg/mL for 8-iso-PGF_2α and 5.0 pg/mL for 8,12-iso-PF_2α-VI for the CSF method and 10.0 pg/0.1 g of tissue and 30.0 pg/0.1 g of tissue for 8-iso-PGF_2α and 8,12-iso-iPF_2α-VI, respectively, for the brain tissue method. No ion suppression or enhancement of the detection of 8-isoPGF_2α, 8,12-isoPF_2α-VI or both internal standards was found.