Comparison of a commercial real-time PCR panel to routine laboratory methods for the diagnosis of meningitis-encephalitis.

Meningitis-encephalitis can range from a mild, self limiting illness to a life threatening disease. Rapid microbial diagnosis allows for early targeted management. This study aimed to compare the BioFire FilmArray Meningitis/Encephalitis multiplex PCR panel (ME panel) to traditional testing algorithms for accuracy and turnaround time in the diagnosis of meningitis-encephalitis. From April to November 2018, cerebrospinal fluid (CSF) samples meeting existing laboratory testing criteria for suspected community acquired meningitis-encephalitis were tested on the ME panel and by routine laboratory methods. The methods were compared for accuracy of diagnosis and turnaround time. Where an organism was not identified, the study investigators came to a consensus on whether an infective aetiology was likely based on CSF parameters, clinical features, management and final discharge diagnosis. A total of 147 CSF samples met criteria for testing. Results were concordant in 143 (97%) of cases, including 27 samples where the same organism was identified by both methods. Of the four discordant samples, three organisms identified by the ME panel alone were considered clinically insignificant. One sample, which was culture and antigen positive for Cryptococcus neoformans, was not detected on the ME panel. The ME panel and routine methods identified an organism in 55% and 58% of clinically compatible cases of infection, respectively. The median turnaround time for the ME panel was 2.9 hours, compared to 21.1 hours for routine testing. The ME panel showed high concordance with traditional testing, simplified laboratory workflow, and significantly reduced turnaround time. The failure of the ME panel to detect Cryptococcus spp. is concerning. When cryptococcal meningitis is suspected, we would recommend using culture and cryptococcal antigen testing as the investigations of choice. Despite the availability of molecular assays targeting the common causes of CNS infection, the diagnostic yield remains suboptimal.

mTORC1 Controls Phase Separation and the Biophysical Properties of the Cytoplasm by Tuning Crowding.

Macromolecular crowding has a profound impact on reaction rates and the physical properties of the cell interior, but the mechanisms that regulate crowding are poorly understood. We developed genetically encoded multimeric nanoparticles (GEMs) to dissect these mechanisms. GEMs are homomultimeric scaffolds fused to a fluorescent protein that self-assemble into bright, stable particles of defined size and shape. By combining tracking of GEMs with genetic and pharmacological approaches, we discovered that the mTORC1 pathway can modulate the effective diffusion coefficient of particles ≥20 nm in diameter more than 2-fold by tuning ribosome concentration, without any discernable effect on the motion of molecules ≤5 nm. This change in ribosome concentration affected phase separation both in vitro and in vivo. Together, these results establish a role for mTORC1 in controlling both the mesoscale biophysical properties of the cytoplasm and biomolecular condensation.

Mucormycosis in Australia: contemporary epidemiology and outcomes.

Mucormycosis is the second most common cause of invasive mould infection and causes disease in diverse hosts, including those who are immuno-competent. We conducted a multicentre retrospective study of proven and probable cases of mucormycosis diagnosed between 2004-2012 to determine the epidemiology and outcome determinants in Australia. Seventy-four cases were identified (63 proven, 11 probable). The majority (54.1%) were caused by Rhizopus spp. Patients who sustained trauma were more likely to have non-Rhizopus infections relative to patients without trauma (OR 9.0, p 0.001, 95% CI 2.1-42.8). Haematological malignancy (48.6%), chemotherapy (42.9%), corticosteroids (52.7%), diabetes mellitus (27%) and trauma (22.9%) were the most common co-morbidities or risk factors. Rheumatological/autoimmune disorders occurred in nine (12.1%) instances. Eight (10.8%) cases had no underlying co-morbidity and were more likely to have associated trauma (7/8; 87.5% versus 10/66; 15.2%; p <0.001). Disseminated infection was common (39.2%). Apophysomyces spp. and Saksenaea spp. caused infection in immuno-competent hosts, most frequently associated with trauma and affected sites other than lung and sinuses. The 180-day mortality was 56.7%. The strongest predictors of mortality were rheumatological/autoimmune disorder (OR = 24.0, p 0.038 95% CI 1.2-481.4), haematological malignancy (OR = 7.7, p 0.001, 95% CI 2.3-25.2) and admission to intensive care unit (OR = 4.2, p 0.02, 95% CI 1.3-13.8). Most deaths occurred within one month. Thereafter we observed divergence in survival between the haematological and non-haematological populations (p 0.006). The mortality of mucormycosis remains particularly high in the immuno-compromised host. Underlying rheumatological/autoimmune disorders are a previously under-appreciated risk for infection and poor outcome.

Case fatality ratio and mortality rate trends of community-onset Staphylococcus aureus bacteraemia.

Lethal outcomes can be expressed as a case fatality ratio (CFR) or as a mortality rate per 100 000 population per year (MR). Population surveillance for community-onset methicillin-sensitive (MSSA) and methicillin-resistant (MRSA) Staphylococcus aureus bacteraemia was conducted in Canada, Australia, Sweden and Denmark to evaluate 30-day CFR and MR trends between 2000 and 2008. The CFR was 20.3% (MSSA 20.2%, MRSA 22.3%) and MR was 3.4 (MSSA 3.1, MRSA 0.3) per 100 000 per year. Although MSSA CFR was stable the MSSA MR increased; MRSA CFR decreased while its MR remained low during the study. Community-onset S. aureus bacteraemia, particularly MSSA, is associated with major disease burden. This study highlights complementary information provided by evaluating both CFR and MR.

The changing epidemiology of Staphylococcus aureus bloodstream infection: a multinational population-based surveillance study.

Although the epidemiology of Staphylococcus aureus bloodstream infection (BSI) has been changing, international comparisons are lacking. We sought to determine the incidence of S. aureus BSI and assess trends over time and by region. Population-based surveillance was conducted nationally in Finland and regionally in Canberra, Australia, western Sweden, and three areas in each of Canada and Denmark during 2000-2008. Incidence rates were age-standardized and gender-standardized to the EU 27-country 2007 population. During 83 million person-years of surveillance, 18,430 episodes of S. aureus BSI were identified. The overall annual incidence rate for S. aureus BSI was 26.1 per 100,000 population, and those for methicillin-sensitive S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA) were 24.2 and 1.9 per 100,000, respectively. Although the overall incidence of community-onset MSSA BSI (15.0 per 100,000) was relatively similar across regions, the incidence rates of hospital-onset MSSA (9.2 per 100,000), community-onset MRSA (1.0 per 100,000) and hospital-onset MRSA (0.8 per 100,000) BSI varied substantially. Whereas the overall incidence of S. aureus BSI did not increase over the study period, there was an increase in the incidence of MRSA BSI. Major changes in the occurrence of community-onset and hospital-onset MSSA and MRSA BSI occurred, but these varied significantly among regions, even within the same country. Although major changes in the epidemiology of community-onset and hospital-onset MSSA and MRSA BSIs are occurring, this multinational population-based study did not find that the overall incidence of S. aureus BSI is increasing.

Anaerobic digestion of corn ethanol thin stillage in batch and by high-rate down-flow fixed film reactors.

Thin stillage (CTS) from a dry-grind corn ethanol plant was evaluated as a carbon source for anaerobic digestion (AD) by batch and high rate semi-continuous down-flow stationary fixed film (DSFF) reactors. Biochemical methane potential (BMP) assays were carried out with CTS concentrations ranging from approximately 2,460-27,172 mg total chemical oxygen demand (TCOD) per litre, achieved by diluting CTS with clean water or a combination of clean water and treated effluent. High TCOD, SCOD and volatile solids (VS) removal efficiencies of 85 ± 2, 94 ± 0 and 82 ± 1% were achieved for CTS diluted with only clean water at an organic concentration of 21,177 mg TCOD per litre, with a methane yield of 0.30 L methane per gram TCOD(removed) at standard temperature and pressure (STP, 0 °C and 1 atmosphere). Batch studies investigating the use of treated effluent for dilution showed promising results. Continuous studies employed two mesophilic DSFF anaerobic digesters treating thin stillage, operated at hydraulic retention times (HRT) of 20, 14.3, 8.7, 6.3, 5 and 4.2 d. Successful digestion was achieved up to an organic loading rate (OLR) of approximately 7.4 g TCOD L(-1)d(-1) at a 5 d HRT with a yield of 2.05 LCH(4) L(-1)d(-1) (at STP) and TCOD and VS removal efficiencies of 89 ± 3 and 85 ± 3%, respectively.

Prolonged carriage of resistant E. coli by returned travellers: clonality, risk factors and bacterial characteristics.

The aim of this study was to delineate the potential risks and dynamics of the prolonged carriage of resistant E. coli in returned travellers. A sample of 274 previously collected E. coli resistant to ceftriaxone (CRO), ciprofloxacin, gentamicin and/or nalidixic acid recovered from 102 travellers was studied. Travellers were assessed pre-travel then longitudinally (maximum 6 months) with peri-rectal/rectal swabs. Clonality was determined by REP-PCR and the presence of O25b-ST131 was assessed. Comparison was made longitudinally for individuals and between identified co-travellers. The risk of prolonged carriage was lower for CRO than for ciprofloxacin or gentamicin resistance. Repeated isolation of the same phenotype at different time points occurred in 19% of initial CRO-resistant carriers compared with 50% of ciprofloxacin- or gentamicin-resistant carriers. The duration of carriage was also longer for the latter resistance phenotypes (75th quartile 8 vs 62 and 63 days respectively). In multivariate analysis, risks of prolonged carriage included antimicrobial use whilst travelling (3.3, 1.3-8.4) and phylogenetic group B2 (9.3, 3.4-25.6) and D (3.8, 1.6-8.8). Clonality amongst longitudinal isolates from the same participant was demonstrated in 92% of participants who were assessable and most marked amongst CRO-resistant isolates. ST-131 was surprisingly infrequent (3% of participants). Prolonged carriage of ciprofloxacin- and gentamicin-resistant isolates is more frequent and prolonged than CRO resistance after travel. Risks of prolonged carriage indicate a contribution of host and bacterial factors to this carriage. These require further elucidation. The strong clonality identified suggests that carriage of a "phenotype" was mediated by persistence of bacteria/plasmid combinations rather than persistence of the plasmid after horizontal transfer to other bacteria.

Serratia sp. bacteremia in Canberra, Australia: a population-based study over 10 years.

The purpose of this paper was to determine the population incidence and clinical features of Serratia sp. bacteremia in Canberra, Australia. Demographic and clinical data were collected prospectively for episodes of Serratia sp. bacteremia over a 10-year period, and was confined to Canberra residents using residential postal codes. Thirty-eight episodes of Serratia sp. bacteremia occurred, with a yearly incidence of 1.03 per 100,000 population. The majority of episodes occurred in males (68%). The respiratory tract was the most common focus of infection (21%). Twenty-nine percent of episodes were community-associated. A further 18% of episodes had their onset in the community but were healthcare-associated. The 7-day and 6-month mortality rates were 5 and 37%, respectively. Antibiotic resistance to gentamicin (3%) and ciprofloxacin (0%) was low. Serratia sp. bacteremia is more common than generally appreciated, with a large proportion (47%) of episodes having their onset in the community.

Initial examination of microwave pretreatment on primary, secondary and mixed sludges before and after anaerobic digestion.

The effects of microwave pretreatment on disintegration and mesophilic digestion of waste activated sludge (WAS), primary sludge (PS), combined (PS + WAS) sequencing batch reactor (SBR) sludge and anaerobically digested biocake were investigated by both household and bench scale industrial types microwaves at temperatures below and above boiling point. Pretreatment variables, temperature, intensity (cooking rate) and sludge concentration had statistically significant effects on solubilization. The microwave pretreatment also increased the bioavailability of sludge components under batch anaerobic digestion and enhanced the dewaterability of pretreated sludges after digestion. However, the level of improvements in solubilization and biodegradation from different waste sludges were different. While the largest improvement in ultimate biodegradation was observed in WAS, microwave irradiation only affected the rate of biodegradation of pretreated PS samples. Similarly, relatively lower solubilization ratios achieved for combined - SBR sludge was attributed to high sludge age of extended aeration SBR unit. It is possible that initial sludge characteristics may influence final pretreatment outcomes so that general statements of performance cannot always be made.

Anaerobic biodegradation of aircraft deicing fluid in UASB reactors.

A central composite design was employed to methodically investigate anaerobic treatment of aircraft deicing fluid (ADF) in bench-scale Upflow Anaerobic Sludge Blanket (UASB) reactors. A total of 23 runs at 17 different operating conditions (0.8% 1.6% ADF (6000-12,000mg/L COD), 12-56h HRT, and 18-36gVSS/L) were conducted in continuous mode. The development of four empirical models describing process responses (i.e. COD removal efficiency, biomass-specific acetoclastic activity, methane production rate, and methane production potential) as functions of ADF concentration, hydraulic retention time, and biomass concentration is presented. Model verification indicated that predicted responses (COD removal efficiencies, biomass-specific acetoclastic activity, and methane production rates and potential) were in good agreement with experimental results. Biomass-specific acetoclastic activity was improved two-fold from 0.23gCOD/gVSS/d for inoculum to a maximum of 0.55gCOD/gVSS/d during ADF treatment in UASB reactors. For the design window, COD removal efficiencies were higher than 90%. The predicted methane production potentials were close to theoretical values, and methane production rates increased as the organic loading rate is increased. ADF toxicity effects were evident for 1.6% ADF at medium organic loadings (SOLR above 0.5gCOD/gVSS/d). In contrast, good reactor stability and excellent COD removal efficiencies were achieved at 1.2% ADF for reactor loadings approaching that of highly loaded systems (0.73gCOD/gVSS/d).

Fed-batch production of citric acid by Candida lipolytica grown on n-paraffins.

This study reports on the effects of fermentor agitation and fed-batch mode of operation on citric acid production from Candida lipolytica using n-paraffin as the carbon source. An optimum range of agitation speeds in the 800-1000 rpm range corresponding to Reynolds numbers of 50000-63000 (based on initial batch conditions) seemed to give the best balance between substrate utilization for biomass growth and citric acid production. Application of multiple fed-batch feedings can be used to extend the batch fermentation and increase final citric acid concentrations and product yield. The three-cycle fed-batch system increased overall citric acid yields to 0.8-1.0 g citricacid/g n-paraffin, approximately a 100% improvement in product yield from those observed in the single cycle fed-batch system and a 200% improvement over normal batch operation. The three-cycle fed-batch mode of operation also increased the final citric acid concentration to 42 g/l from about 12 and 6g/l for single fed-batch cycle and normal batch modes of operation, respectively. Increased citric acid concentrations in three-cycle fed-batch mode was achieved at longer fermentation times.

Super blue box recycling (SUBBOR) enhanced two-stage anaerobic digestion process for recycling municipal solid waste: laboratory pilot studies.

The super blue box recycling (SUBBOR) process is an enhanced, multi-stage anaerobic digestion process for mixed municipal solid waste (MSW) and other biomass feedstock materials. The technology centers on enhanced high solids, thermophilic digestion after steam-pressure disruption of the ligno-cellulosic fiber components that are recalcitrant to conventional anaerobic digestion. Mixed MSW, rich in organic components but also containing inorganic materials, such as glass, aluminum and steel, as well as non-digestible plastic materials, has been laboratory pilot tested with a fully integrated process train designed to treat and recycle all of the MSW components. Methane yields from the MSW were 0.36 m3 CH4/kg volatile solids (VS) representing a 40% increase over the yield obtained from conventional single stage digestion. The secondary digestion step after steam pressure disruption also provided a 40% improvement in total solids and VS reduction. The residual organic fraction following two-stage digestion was fine in texture and was recovered as a clean peat fraction with reduced contents of heavy metal and other fugitive non-digested contaminants. Mass and energy balance determinations indicated a high degree of MSW diversion from landfill disposal (>80%) was achievable by the SUBBOR process as well as substantial net electrical and thermal energy production. Continuous long-term trials of the SUBBOR process at 25,000 tonnes/year are underway.

Synthesis and analysis of potent, more lipophilic derivatives of the bradykinin B2 receptor antagonist peptide Hoe 140.

Bradykinin (BK) is an endogenous peptide that has been implicated in several pathological conditions, hence antagonists of its activity have therapeutic potential. The decapeptide Hoe 140 is currently one of the best BK antagonists, but interest remains in finding even more potent compounds. A library of Hoe 140 derivatives was synthesized that incorporated non-natural analogs of the cationic, naturally occurring amino acids arginine (Arg) and lysine (Lys). The modified amino acids were designed to form enhanced ionic interactions due to an increase in local hydrophobicity, which promotes desolvation of the cation in water. The potencies of the resulting peptides were determined by competitive binding assays in human A431 cells expressing the BK B2 receptor. Two of the peptides synthesized were equipotent to Hoe 140 (IC(50s) 2.99 and 3.36 nM) and the most potent was demonstrated as a functional antagonist in vitro by blocking BK-mediated phosphorylation of mitogen-activated protein (MAP) kinases. The new derivatives are more hydrophobic than Hoe 140 and thus may exhibit changes in pharmacokinetic properties when evaluated in vivo.

Neutral fat hydrolysis and long-chain fatty acid oxidation during anaerobic digestion of slaughterhouse wastewater.

Neutral fat hydrolysis and long-chain fatty acid (LCFA) oxidation rates were determined during the digestion of slaughterhouse wastewater in anaerobic sequencing batch reactors operated at 25 degrees C. The experimental substrate consisted of filtered slaughterhouse wastewater supplemented with pork fat particles at various average initial sizes (D(in)) ranging from 60 to 450 microm. At the D(in) tested, there was no significant particle size effect on the first-order hydrolysis rate. The neutral fat hydrolysis rate averaged 0.63 +/- 0.07 d(-1). LCFA oxidation rate was modelled using a Monod-type equation. The maximum substrate utilization rate (kmax) and the half-saturation concentration (Ks) averaged 164 +/- 37 mg LCFA/L/d and 35 +/- 31 mg LCFA/L, respectively. Pork fat particle degradation was mainly controlled by LCFA oxidation rate and, to a lesser extent, by neutral fat hydrolysis rate. Hydrolysis pretreatment of fat-containing wastewaters and sludges should not substantially accelerate their anaerobic treatment. At a D(in) of 450 microm, fat particles were found to inhibit methane production during the initial 20 h of digestion. Inhibition of methane production in the early phase of digestion was the only significant effect of fat particle size on anaerobic digestion of pork slaughterhouse wastewater. Soluble COD could not be used to determine the rate of lipid hydrolysis due to LCFA adsorption on the biomass.

Differential synthesis of peritoxins and precursors by pathogenic strains of the fungus Periconia circinata.

Pathogenic strains of the soilborne fungus Periconia circinata produce peritoxins with host-selective toxicity against susceptible genotypes of sorghum. The peritoxins are low-molecular-weight, hybrid molecules consisting of a peptide and a chlorinated polyketide. Culture fluids from pathogenic, toxin-producing (Tox(+)) and nonpathogenic, non-toxin-producing (Tox(-)) strains were analyzed directly by gradient high-performance liquid chromatography (HPLC) with photodiode array detection and HPLC-mass spectrometry to detect intermediates and final products of the biosynthetic pathway. This approach allowed us to compare the metabolite profiles of Tox(+) and Tox(-) strains. Peritoxins A and B and the biologically inactive intermediates, N-3-(E-pentenyl)-glutaroyl-aspartate, circinatin, and 7-chlorocircinatin, were detected only in culture fluids of the Tox(+) strains. The latter two compounds were produced consistently by Tox(+) strains regardless of the amount of peritoxins produced under various culture conditions. In summary, none of the known peritoxin-related metabolites were detected in Tox(-) strains, which suggests that these strains may lack one or more functional genes required for peritoxin biosynthesis.

Optimization of citric acid production from Candida lipolytica Y-1095 using n-paraffin.

Currently, the majority of worldwide microbial production of citric acid utilizes Aspergillus niger in a carbohydrate based submerged fermentation. Due to their high carbon content, hydrocarbons also have the potential of producing high concentrations of citric acid. Initial lab experiments conducted using 1875 ml batch fermentations with n-paraffin found that Candida lipolytica NRRL-Y-1095 assimilated the feedstock and had a citric acid productivity of 47 mg l(-1) h(-1). To determine the optimum level of initial biomass concentration, n-paraffin concentration, iron concentration and temperature for the production of citric acid, a central composite design was developed using 200 ml batch fermentations. The design involved conducting 31 batch fermentations under various combinations of high and low values of these four parameters. From this investigation empirical models were developed describing the interactions between the experimental parameters and citric acid production. It was found that the maximum concentration of citric acid produced was 9.8 g l(-1) and the optimum levels of each parameter for citric acid production were, 10--12% volume for initial biomass concentration, 10--15% volume for n-paraffin concentration, 10 mg l(-1) for ferric nitrate concentration, and 26--30 degrees C for temperature.

CXCL10 (IFN-gamma-inducible protein-10) control of encephalitogenic CD4+ T cell accumulation in the central nervous system during experimental autoimmune encephalomyelitis.

Experimental autoimmune encephalomyelitis (EAE) is a CD4(+) Th1-mediated demyelinating disease of the CNS that serves as a model for multiple sclerosis. A critical event in the pathogenesis of EAE is the entry of both Ag-specific and Ag-nonspecific T lymphocytes into the CNS. In the present report, we investigated the role of the CXC chemokine CXCL10 (IFN-gamma-inducible protein-10) in the pathogenesis of EAE. Production of CXCL10 in the CNS correlated with the development of clinical disease. Administration of anti-CXCL10 decreased clinical and histological disease incidence, severity, as well as infiltration of mononuclear cells into the CNS. Anti-CXCL10 specifically decreased the accumulation of encephalitogenic PLP(139-151) Ag-specific CD4+ T cells in the CNS compared with control-treated animals. Anti-CXCL10 administration did not affect the activation of encephalitogenic T cells as measured by Ag-specific proliferation and the ability to adoptively transfer EAE. These results demonstrate an important role for the CXC chemokine CXCL10 in the recruitment and accumulation of inflammatory mononuclear cells during the pathogenesis of EAE.

Conformational changes in Leishmania mexicana glyceraldehyde-3-phosphate dehydrogenase induced by designed inhibitors.

The glycolytic enzymes of trypanosomes are attractive drug targets, since the blood-stream form of Trypanosoma brucei lacks a functional citric acid cycle and is dependent solely on glycolysis for its energy requirements. Glyceraldehyde-3-phosphate dehydrogenases (GAPDH) from the pathogenic trypanosomatids T. brucei, Trypanosoma cruzi and Leishmania mexicana are quite similar to each other, and yet have sufficient structural differences compared to the human enzyme to enable the structure-based design of compounds that selectively inhibit all three trypanosomatid enzymes but not the human homologue. Adenosine analogs with substitutions on N-6 of the adenine ring and on the 2' position of the ribose moiety were designed, synthesized and tested for inhibition. Two crystal structures of L. mexicana glyceraldehyde-3-phosphate dehydrogenase in complex with high-affinity inhibitors that also block parasite growth were solved at a resolution of 2.6 A and 3.0 A. The complexes crystallized in the same crystal form, with one and a half tetramers in the crystallographic asymmetric unit. There is clear electron density for the inhibitor in all six copies of the binding site in each of the two structures. The L. mexicana GAPDH subunit exhibits substantial structural plasticity upon binding the inhibitor. Movements of the protein backbone, in response to inhibitor binding, enlarge a cavity at the binding site to accommodate the inhibitor in a classic example of induced fit. The extensive hydrophobic interactions between the protein and the two substituents on the adenine scaffold of the inhibitor provide a plausible explanation for the high affinity of these inhibitors for trypanosomatid GAPDHs.

Testing of alkaline and enzymatic hydrolysis pretreatments for fat particles in slaughterhouse wastewater.

Four pretreatments to hydrolyse and/or reduce the size of fat particles in slaughterhouse wastewater (SHW) were tested: sodium hydroxide and three lipases of plant, bacterial and animal (pancreatic) origin. Hydrolysing agents and SHW containing between 2.5 and 3 g/l of fat particles were mixed at room temperature for 4 h. Additions of 5-400 meq NaOH/l did not increase soluble COD (SCOD) in SHW, but the average particle size was reduced to 73% +/- 7% of the initial average particle size (D(in)) at NaOH concentrations ranging from 150 to 300 meq/l. Pretreatment with pancreatic lipase PL-250 reduced the average particle size to a maximum of 60% +/- 3% of D(in). As D(in) was decreased from 359 to 68 microns, the enzyme concentration required to obtain the maximum particle size reduction increased from 200 to 1000 mg/l. A 4-h pretreatment with PL-250 also increased the free long-chain fatty acid (LCFA) concentration to a maximum of 15.5 mg/l, indicating some solubilization of the pork fat particles in SHW. SCOD was not significantly increased by the pretreatment, but SCOD was not found to be a good indicator of enzymatic lipolysis because of enzyme adsorption on the fat particle surface. Pancreatic lipase appeared more efficient with beef fat than pork fat, possibly because beef fat contains less polyunsaturated fatty acids than pork fat. The bacterial lipase LG-1000 was also efficient in reducing average fat particle size, but high doses (> 1000 mg/l) were required to obtain a significant reduction after 4 h of pretreatment. SCOD was not increased by pretreatment with LG-1000. No particle size reduction or changes in SCOD were noted after 4 h of pretreatment with the plant lipase EcoSystem Plus. It was concluded that PL-250 was the best pretreatment to hydrolyse fat particles in SHW. However, its impact on the efficiency of a downstream anaerobic digestion process remains to be tested.

A disubstituted NAD+ analogue is a nanomolar inhibitor of trypanosomal glyceraldehyde-3-phosphate dehydrogenase.

N6-Naphthalenemethyl-2'-methoxybenzamido-beta-NAD+, a derivative of a low micromolar first-generation inhibitor of trypanosomal glyceraldehyde phosphate dehydrogenase (GAPDH), was synthesized, taking advantage of methodology for the selective phosphitylation of nucleosides. The compound was found to be a poor alternate cosubstrate for GAPDH, but an extremely potent inhibitor. Although intended for use in crystallization trials, the analogue presents possibilities for further drug design.