Changes in volatile fatty acid production and microbiome during fermentation of food waste from hospitality sector.

Due to the increasing demand for low carbon-footprint bioproducts in the markets, innovative processes technologies and products are needed. The objective of this study was to assess the quality and potential of food waste (FW) from the hospitality sector to produce volatile fatty acids (VFAs). A batch type acid fermentation system was used to study VFA production in different process conditions (a decreased pH and increased organic loading rate). The evolution of VFAs and long-chain fatty acids was followed. Amplicon sequencing of the 16S rRNA gene was used to investigate the bacterial and archaeal community, and elucidate microbial communities in different FW and process conditions. The results show that high VFA concentrations (of up to 18 g/L) were achieved in overloaded conditions, which were also affected by the activity and composition of the inoculum. FW played an important role in modulating microbial composition, especially the bacterial communities belonging to the lactic acid bacteria group.

Ultrafast transient liquid assisted growth of high current density superconducting films.

The achievement of high growth rates in YBa2Cu3O7 epitaxial high-temperature superconducting films has become strategic to enable high-throughput manufacturing of long length coated conductors for energy and large magnet applications. We report on a transient liquid assisted growth process capable of achieving ultrafast growth rates (100 nm s-1) and high critical current densities (5 MA cm-2 at 77 K). This is based on the kinetic preference of Ba-Cu-O to form transient liquids prior to crystalline thermodynamic equilibrium phases, and as such is a non-equilibrium approach. The transient liquid-assisted growth process is combined with chemical solution deposition, proposing a scalable method for superconducting tapes manufacturing. Additionally, using colloidal solutions, the growth process is extended towards fabrication of nanocomposite films for enhanced superconducting properties at high magnetic fields. Fast acquisition in situ synchrotron X-ray diffraction and high resolution scanning transmission electron microscopy (STEM) become crucial measurements in disentangling key aspects of the growth process.

Preimplantation genetic diagnosis for chromosome rearrangements - one blastomere biopsy versus two blastomere biopsy.

PURPOSE: Preimplantation Genetic Diagnosis (PGD) has proven to be a useful reproductive option for carriers of some chromosome rearrangements. The data presented in this study compares the impact of one versus two blastomere biopsy on the likelihood of achieving a PGD result, as well as the effect on subsequent embryo development and clinical outcomes. METHODS: IVF-PGD couples had either one or two blastomeres biopsied from all embryos with ≥7 blastomeres on day 3 post oocyte collection. These blastomeres were assessed for the specific chromosome rearrangement using Fluorescent In-situ Hybridisation (FISH). Further embryo development was monitored on days 4 and 5. Clinical outcomes were assessed retrospectively. RESULTS: The data shows that statistically more embryos achieved a PGD result following two blastomere biopsy, compared with one blastomere biopsy (92 % versus 88 %, respectively). Furthermore it was found that embryo development and clinical outcomes were similar between the two biopsy groups. CONCLUSIONS: Based on this analysis it appears that the biopsy of two blastomeres from embryos with ≥7 blastomeres on day 3 is a valid and successful approach for couples presenting for IVF-PGD for a chromosome rearrangement.

The genome of chemorefractory chronic lymphocytic leukemia reveals frequent mutations of NOTCH1 and SF3B1.

Next-generation whole-exome sequencing has revealed two novel genes, namely NOTCH1 and SF3B1, whose mutations predict poor outcome and preferentially associate with chemorefractory chronic lymphocytic leukemia (CLL). Analysis of 539 CLL cases documents that NOTCH1 mutations i) represent one of the most frequent cancer gene mutations involved at presentation; ii) cluster with cases harboring trisomy 12 and tend to be mutually exclusive with TP53 disruption among genetic subgroups; iii) identify high-risk patients showing poor survival similar to that associated with TP53 abnormalities; and iv) exert a prognostic role independent of widely accepted clinical and genetic risk factors. Mutations of SF3B1, a splicing factor that is a critical component of the spliceosome, recurrently associate with fludarabine-refractory CLL, occur at a low rate at CLL presentation and have a minor role in Richter transformation, corroborating the notion that CLL histological shift is molecularly distinct from chemorefractory progression without the Richter transformation.

Analysis of the host pharmacogenetic background for prediction of outcome and toxicity in diffuse large B-cell lymphoma treated with R-CHOP21.

Knowledge on the impact of pharmacogenetics in predicting outcome and toxicity in diffuse large B-cell lymphoma (DLBCL) is scant. We tested 106 consecutive DLBCL treated with R-CHOP21 for 19 single nucleotide polymorphisms (SNPs) from 15 genes potentially relevant to rituximab-CHOP (R-CHOP) pharmacogenetics. Associations of SNPs with event-free survival (EFS) and toxicity were controlled for multiple testing. Genotypic variants of nicotinamide adenine dinucleotide phosphate (NAD(P)H) oxidase p22phox (CYBA rs4673) and alpha1 class glutathione S-transferase (GSTA1 rs3957357) were independent predictors of EFS (CYBA rs4673 TT genotype: HR 2.06, P=0.038; GSTA1 rs3957357 CT/TT genotypes: HR 0.38, P=0.003), after adjusting for International Prognostic Index (IPI). CYBA rs4673 and GSTA1 rs3957357 also predicted outcome in DLBCL subgroups by IPI. Impact of SNPs on toxicity was evaluated in 658 R-CHOP21 courses utilizing generalized estimating equations. NCF4 rs1883112 was an independent predictor against hematologic (odds ratios (OR): 0.45; P=0.018), infectious (OR: 0.46; P=0.003) and cardiac toxicity (OR: 0.37; P=0.023). Overall, host SNPs affecting doxorubicin pharmacodynamics (CYBA rs4673) and alkylator detoxification (GSTA1 rs3957357) may predict outcome in R-CHOP21-treated DLBCL. Also, NCF4 rs1883112, a SNP of NAD(P)H oxidase p40phox, may have a function in protecting against hematologic and nonhematologic toxicity. These results highlight the need to improve characterization of the host genetic background for a better prognostication of DLBCL.

Landfill gas upgrading with countercurrent water wash.

A pilot-scale countercurrent absorption process for upgrading landfill gas to produce vehicle fuel was studied using absorption and desorption units and water as absorbent. The height-to-diameter ratio of the absorption column used was 3:1 instead of the more conventionally used 20:1 ratio, and a higher pressure was used along with a lower water flow rate. The effects of pressure (10-30 bar) and water and gas flow-ratios on the upgrading process were studied. Methane content in the product gas increased to near or above 90% with both gas flows (50 and 100 l/min) used at over 20 bar pressure with 10 l/min water flow and at 30 bar pressure with 5l/min water flow. Carbon dioxide content with these upgrading parameters ranged from 3.2% to 4.8%. The remaining fraction of the product gas was nitrogen (from 6% to 7%), while hydrogen sulphide was removed to below the detection limit with all of the upgrading parameters used. The methane content of exhaust gas increased with increasing pressure. In conclusion, the pilot-scale gas upgrading process studied here appears to be able to produce gas with high energy content (>90% methane), apparently suitable as vehicle fuel, from landfill gas.

Protein metabolism and strength performance after bovine colostrum supplementation.

This study was designed to determine the responses of muscle protein, serum amino acids, and strength performance to bovine colostrum supplementation in physically active men. The rest (R) group (n = 6) and the exercise (E) group (n = 6) carried out twice a 2-week experiment randomly assigned in a double-blind fashion with either placebo (PLA; consuming daily 20 g maltodextrin) or bovine colostrum (COL; consuming daily 20 g colostrum supplement) treatment with one month between. On the test day after the treatment period the measurements were carried out in fasting conditions and E carried out a strength training session (STS). The methods involved the infusion of ring-(2)H(5)-phenylalanine, femoral arterial and venous blood sampling, and biopsies from the vastus lateralis muscle. Serum concentration of essential amino acids during recovery was greater (p < 0.05) in the COL groups compared with the PLA groups. Both muscle protein synthesis and breakdown increased (p < 0.05) with COL. There were no differences in phenylalanine net balance or strength performance between the PLA and COL groups. It was concluded that a 2-week supplementation with bovine colostrum in physically active men increases serum concentration of essential amino acids but has no effect either on strength performance or protein net balance in fasting conditions during recovery after STS.

Rotaviral antibodies in the treatment of acute rotaviral gastroenteritis.

The efficacy of hyperimmune bovine colostrum received from cows immunized with simian rotavirus SA11 in the treatment of rotavirus gastroenteritis was compared in a randomized double-blind trial to colostrum and ordinary milk preparations. One hundred and thirty-five children aged 6-30 months with rotaviral gastroenteritis received either hyperimmune bovine colostrum (n=42), ordinary colostrum (n=42) or milk (n=41) as a 100 ml solution four times/d for 4 d. Even though the differences were in favour of hyperimmune bovine colostrum in all the variables evaluated [greater weight gain (403 vs 343 g), shorter duration of diarrhoea (3.1 vs 3.6 d), fewer stools during 6 d (11.5 vs 13.6) and fewer stools during the first 3 d (9.3 vs 11.3)], all the differences were statistically insignificant. Differences of this size are clinically unimportant in well-nourished immunocompetent children, but we suggest that the hyperimmune bovine colostrum tested in our trial had some effects in the treatment of acute rotaviral gastroenteritis and should be evaluated further.

Quantification of carbonic anhydrase III and myoglobin in different fiber types of human psoas muscle.

Carbonic anhydrase (CA III) and myoglobin contents from isolated human muscle fibers were quantified using a sensitive time-resolved fluoroimmunoassay. Human psoas muscle specimens were freeze-dried, and single fibers were dissected out and classified into type I, IIA and IIB by myosin ATPase staining. Fiber typing was further confirmed by SDS-PAGE. CA III and myoglobin were found in all fiber types. Type I fibers contained higher concentrations of CA III and myoglobin than type IIA and IIB fibers. The relative concentrations of CA III in type IIA and IIB fibers were respectively 24% and 10% of that in type I fibers. The relative concentrations of myoglobin in type IIA and IIB fibers were 60% and 28% of that in type I fibers. Anti-CA III immunoblotting results from fiber-specific pooled samples agreed well with quantitative measurements. The results indicate that CA III is a more specific marker than myoglobin for type I fibers.

Dual-label time-resolved fluoroimmunoassay for simultaneous detection of myoglobin and carbonic anhydrase III in serum.

We developed a dual-labeled time-resolved fluoroimmunoassay for simultaneous quantification of myoglobin (Mb) and carbonic anhydrase III (CA III) in serum involving polyclonal antibodies and the fluorescent lanthanides europium (Eu3+) and samarium (Sm3+). This solid-phase immunoassay is based on competition between Eu(3+)- or Sm(3+)-labeled antigen and the sample antigen for polyclonal rabbit antibodies. Standards and patients' samples containing antigen inhibit binding of the lanthanide-labeled antigen to the antibody. A second antibody directed against rabbit IgG is coated on a solid phase and binds the IgG-antigen-lanthanide complex, giving rapid and complete separation of antibody-bound and free antigen. The assay requires only one incubation step. An enhancement solution dissociates Eu3+ and Sm3+ ions from the labeled CA III and Mb, respectively, into a solution where they form highly fluorescent chelates. Spectra of the fluorescent chelates in the microtitration-strip wells were run on a time-resolved fluorometer equipped with filters for Eu3+ (613 nm) and Sm3+ (643 nm), the fluorescence from each sample being inversely proportional to the concentration of antigens. The measurement range for both analytes is from 5 to 1500 micrograms/L. The mean within- and between-assay precisions (CV) were 4.6% and 6.2% for CA III and 5.9% and 7.3% for Mb, respectively. Good correlations were obtained with the results of CA III RIA and a commercial myoglobin RIA kit.

Time-resolved fluoroimmunoassay of 5-methyl-2'-deoxycytidine employing europium-labeled antigen as tracer.

We describe a solid-phase fluoroimmunoassay, based on competition between europium-labeled 5 MeCyd (5-methylcytidine) and sample 5MedCyd (5-methyl-2'-deoxycytidine) for polyclonal anti-5MedCyd antibodies (rabbit). Europium labeling of antigen was performed using a novel polylysine-5MeCyd conjugate. Standard and sample preparations containing 5MedCyd inhibited the binding of the europium-labeled 5MeCyd to the antibody molecules. A second antibody, directed against rabbit IgG, was coated on the solid phase, and bound the IgG-5MeCyd-polylysine-europium complex, giving rapid and complete separation of antibody-bound and free antigen. The measuring range was from 3.7 to 2500 pmol of 5MedCyd per assay. A good correlation between the results obtained with TR-FIA and HPLC was demonstrated when the methods were applied to the measurement of methylation in various DNA samples, enzymatically hydrolyzed to their constituent deoxyribonucleosides. This new TR-FIA possesses the same advantages (high sensitivity, wide assay range, rapidity, simplicity, and low cost) as the previous assay developed in our laboratories. The superiority of the new system is based on (i) its low inter- and intra-assay variation, (ii) low antiserum consumption, and (iii) a protocol, which permits the use of second-antibody-coated microtitration strips common to other assays.

Radioimmunoassay of 5-hydroxymethyl-2'-deoxyuridine.

5-Hydroxymethyl-2'-deoxyuridine is an antileukemic thymidine analogue. It is also a well known thymidine-derivative in DNA exposed to ionizing irradiation. We report the production and characterization of specific rabbit anti-5HmdUrd antisera. The antisera were used for the radioimmunological measurement of 5HmdUrd. The radioimmunoassay was capable of quantitating 2 pmol of 5 HmdUrd per tube corresponding to 0.2 mumol/l in a 10 microliter plasma sample. A good correlation between the results obtained with the radioimmunoassay and HPLC was demonstrated when the methods were applied to the measurement of plasma levels of 5HmdUrd in mice receiving experimental chemotherapy.

Time-resolved fluoroimmunoassay of 5-methyl-2'-deoxycytidine.

We describe time-resolved fluoroimmunoassay of 5-methyl-2'-deoxycytidine (5MedCyd). The assay is based on the use of a highly specific antiserum raised in rabbits against BSA-conjugated 5-methylcytidine (5MeCyd). The tracer in the solid-phase time-resolved fluoroimmunoassay (TR-FIA) was antigen-selected anti-5MedCyd labeled with Europium. Thyroglobulin-linked 5MeCyd served as the solid-phase antigen. The measuring range for the fluoroimmunoassay was from less than 1 to 5000 pmol per assay of 5MedCyd. A good correlation between the results obtained with the TR-FIA and HPLC was demonstrated when the methods were applied to the measurement of methylation in human leukemic cells and other DNA samples. TR-FIA has several advantages over the more laborious techniques available so far: (i) high sensitivity, (ii) large assay ranges, (iii) rapidity and large number of simultaneous assays, (iv) simplicity, and (v) low cost provided that the laboratory has equipment for time-resolved fluorometry.

An improved radioimmunoassay for the quantitation of DNA methylation.

A novel radioimmunoassay of 5MedCyd is described. The assay, employing a highly specific antiserum raised in rabbits against BSA-conjugated 5MeCyd, used 5-125iodo-2'-deoxycytidine as the tracer. The measuring range for the assay was found to be 1-1000 pmol per assay of 5MedCyd. When the methods were applied to the measurement of methylation in DNA samples a good correlation between the results obtained with the radioimmunoassay and HPLC was demonstrated. The method has several advantages over the more laborious and sophisticated techniques previously available: high sensitivity, large assay range, rapidity and potential for large number of simultaneous assays, simplicity, and low cost provided that the laboratory has equipment for gamma counting.

Radioimmunoassay of 5-methyl-2'-deoxycytidine. A method for the quantitation of DNA methylation.

5-Methyl-2'-deoxycytidine (5MedCyd) is a minor constituent of mammalian cell DNA. We report the production and characterization of highly specific rabbit anti-5MedCyd antiserum. The antiserum was suitable for the radioimmunological measurement of 5MedCyd. This simple radioimmunoassay was capable of quantitating calf thymus DNA methylation at nanomolar levels of total DNA.