Disease-related amyloidogenic variants of human lysozyme trigger the unfolded protein response and disturb eye development in Drosophila melanogaster

We have created a Drosophila model of lysozyme amyloidosis to investigate the in vivo behavior of disease-associated variants. To achieve this objective, wild-type (WT) protein and the amyloidogenic variants F57I and D67H were expressed in Drosophila melanogaster using the UAS-gal4 system and both the ubiquitous and retinal expression drivers Act5C-gal4 and gmr-gal4. The nontransgenic w(1118) Drosophila line was used as a control throughout. We utilized ELISA experiments to probe lysozyme protein levels, scanning electron microscopy for eye phenotype classification, and immunohistochemistry to detect the unfolded protein response (UPR) activation. We observed that expressing the destabilized F57I and D67H lysozymes triggers UPR activation, resulting in degradation of these variants, whereas the WT lysozyme is secreted into the fly hemolymph. Indeed, the level of WT was up to 17 times more abundant than the variant proteins. In addition, the F57I variant gave rise to a significant disruption of the eye development, and this correlated to pronounced UPR activation. These results support the concept that the onset of familial amyloid disease is linked to an inability of the UPR to degrade completely the amyloidogenic lysozymes prior to secretion, resulting in secretion of these destabilized variants, thereby leading to deposition and associated organ damage.     

Simultaneous Non-Negative Matrix Factorization for Multiple Large Scale Gene Expression Datasets in Toxicology

Non-negative matrix factorization is a useful tool for reducing the dimension of large datasets. This work considers simultaneous non-negative matrix factorization of multiple sources of data. In particular, we perform the first study that involves more than two datasets. We discuss the algorithmic issues required to convert the approach into a practical computational tool and apply the technique to new gene expression data quantifying the molecular changes in four tissue types due to different dosages of an experimental panPPAR agonist in mouse. This study is of interest in toxicology because, whilst PPARs form potential therapeutic targets for diabetes, it is known that they can induce serious side-effects. Our results show that the practical simultaneous non-negative matrix factorization developed here can add value to the data analysis. In particular, we find that factorizing the data as a single object allows us to distinguish between the four tissue types, but does not correctly reproduce the known dosage level groups. Applying our new approach, which treats the four tissue types as providing distinct, but related, datasets, we find that the dosage level groups are respected. The new algorithm then provides separate gene list orderings that can be studied for each tissue type, and compared with the ordering arising from the single factorization. We find that many of our conclusions can be corroborated with known biological behaviour, and others offer new insights into the toxicological effects. Overall, the algorithm shows promise for early detection of toxicity in the drug discovery process.     

Olive oil history,production and by-product management

In this review paper, the history of the olive tree and the development of the main olive oil extraction systems employed in the past are presented. Furthermore, the management techniques employed from antiquity until today for the treatment of olive mill wastewater (OMW), which constitutes the most important by-product of olive oil production, are addressed and compared. Reference is also made to global olive oil production, the environmental impacts of the uncontrolled disposal of OMW, and to the legal framework concerning its management in the main olive oil producing countries. Other by-products of olive processing are also discussed.     

Foraging mechanics and their outcomes for cattle grazing reproductive tropical swards

For grazing animals an important determinant of animal performance is the rate of nutrient intake (RNI) which depends on diet quality and instantaneous intake rate (IIR). In turn, diet quality and IIR are the outcome of the interaction between the morphology and behaviour of the animal and the structure of the sward. Using artificial microswards of Panicum maximum we evaluated the effect of four levels of the tensile resistance of stems in tropical swards on the grazing behaviour of cattle of two age classes (1- and 3-year-old steers) faced with a stem barrier either in a vertical (experiment 1, E1) or horizontal plane (experiment 2, E2). The animals did not select against low tensile resistance stems (LTRS) but did avoid high tensile resistance stems (HTRS) which resulted in a significant difference (P < 0.001) between diet DM (dry matter) digestibility and forage DM digestibility in swards with HTRS. IIR decreased (P < 0.001) on average 62% in E1 and 67% in E2 as stem tensile resistance increased, 144% and 177% in E1 and E2, respectively. This led to a reduction (P < 0.001) in digestible DM IIR in both young and mature cattle respectively of 56% and 68% in E1, and 45% and 79% in E2, as stem tensile resistance increased. The decline in IIR was due to an increase in time per bite and a reduction in bite dimensions that were the result of different mechanisms in the two experiments: when leaves were taller than the stems (E1) the decrease in bite area was associated with an asymptotic increase (P < 0.001) in bite force in mature animals, and a maintenance (P = 0.456) of bite force in young animals, suggesting that maximum bite force regulated bite area in E1. This was not the case in E2 because both bite area and bite force decreased (P < 0.05) with the increase in stem tensile resistance suggesting that the animals were not able to gather enough plant material with the tongue in order for bite force to regulate bite area. We conclude that bite force and tongue force regulate selectivity and bite dimensions, which are the ultimate determinants of the RNI; IIR is the primary determinant of the RNI with diet quality being of lesser importance; and HTRS act as deterrents to achieving maximum IIR in tropical swards, particularly in mature cattle.     

Evaluation of a trapping ELISA for the differentiation of foot-and-mouth disease virus strains using monoclonal antibodies.

A trapping enzyme-linked immunosorbent assay (ELISA) has been evaluated for the differentiation of foot-and-mouth disease virus (FMDV) strains using a panel of seven anti-serotype O monoclonal antibodies (MAbs). The variation of results within and between tests performed on the same day and on different days was examined using three strains of FMDV. Criteria for establishing antigenic differences between the strains as defined by the individual MAbs are proposed based on the variability measured, which can be used as standards by workers performing this test with other MAbs and FMDV strains.     

A different kind of day hospital--for patients with preterminal cancer and chronic disease.

A new day hospital has been set up in Sheffield for patients with preterminal cancer and chronic disease. During the first 26 months 197 patients with cancer and 66 chronically sick patients attended. Of the 2701 attendances by patients with cancer, breast cancer accounted for 38%. One hundred and forty of the patients with cancer died, 83% in the terminal-care unit and 12% at home. Ninety per cent of respondents to a questionnaire thought that the support provided was of great importance to both patient and family; and over two-thirds of the patients were said to have benefited from improved control of symptoms. A similar facility could be developed within existing day hospitals without major revenue consequences, and should produce a genuine improvement in care. This study suggests, however, that it would not shorten the terminal stay in hospital.     

Combination Chemotherapy in Generalized Hodgkin's Disease

Fifty-two patients with generalized Hodgkin''s disease were treated with a combination of mustine hydrochloride, vinblastine, procarbazine, and prednisolone. Complete remissions were obtained initially in six out of seven patients (86%) who had previously received no treatment, in 15 out of 19 (79%) who had had only radiotherapy in the past, and in 9 out of 26 (35%) who had previously been given chemotherapy with or without radiotherapy. Of these 30 patients in whom a complete remission was obtained 22 have been free of any symptoms or signs of disease for periods ranging from 4 to 22 months. The response to treatment was rapid, and toxicity was not a major problem, except in those who had previously been treated with cytotoxic drugs used continuously and not in courses. A comparative trial of radiotherapy and combination therapy in the treatment of Stage III Hodgkin''s disease is strongly recommended.     

Human β-Synuclein Rendered Fibrillogenic by Designed Mutations

Filamentous inclusions made of α-synuclein are found in nerve cells and glial cells in a number of human neurodegenerative diseases, including Parkinson disease, dementia with Lewy bodies, and multiple system atrophy. The assembly and spreading of these inclusions are likely to play an important role in the etiology of common dementias and movement disorders. Both α-synuclein and the homologous β-synuclein are abundantly expressed in the central nervous system; however, β-synuclein is not present in the pathological inclusions. Previously, we observed a poor correlation between filament formation and the presence of residues 73–83 of α-synuclein, which are absent in β-synuclein. Instead, filament formation correlated with the mean β-sheet propensity, charge, and hydrophilicity of the protein (global physicochemical properties) and β-strand contiguity calculated by a simple algorithm of sliding averages (local physicochemical property). In the present study, we rendered β-synuclein fibrillogenic via one set of point mutations engineered to enhance global properties and a second set engineered to enhance predominantly β-strand contiguity. Our findings show that the intrinsic physicochemical properties of synucleins influence their fibrillogenic propensity via two distinct but overlapping modalities. The implications for filament formation and the pathogenesis of neurodegenerative diseases are discussed.     

Prediction of the performance of inbred lines derived from a population cross in autumn-sown onions (Allium cepa L.)

Summary A design and model are presented to allow the prediction, in early generations, of the mean and distribution of recombinant inbred lines derived from a cross between two parental populations or partially inbred lines. The procedure has been tested in autumn-sown onions (in the UK) using a wide cross between the openpollinated Japanese cultivar, Senshyu, and a partially inbred line derived from the European cultivar, Rawska. The early generations used for prediction included the first self-pollinated generation of the two parental populations and the F₃ generation produced from the hybrid population. The predictions were tested by reference to the field performance of a random array of inbred lines, which were produced by single-seed descent (SSD) and had been selfed for three generations. The early generations, used for prediction, and a sample of SSD lines were raised alongside each other in each of two seasons. Within each season, good agreement was found between the predicted and observed performance of the recombinant inbred lines for three characters — yield, quality and maturity. This is used as evidence of the validity of the genetical model and the assumptions made. The effects of genotype x environment interactions prevented predictions made in one season being reliably applied to those made in the other and, therefore, reduce the attraction of this type of prediction study to the plant breeder.     

Three-day and ten-day chemotherapy for urinary tract infections in general practice.

The length of a course of antibiotic treatment for urinary tract infection varies with the habits of the prescriber. Many patients do not complete a course of treatment once their symptoms have subsided. In uncomplicated urinary tract infection among women seen in general practice a three-day course of amoxycillin was as effective as a 10-day course of the same drug in the same dose. Relief of symptoms was equal in both groups and bacteriuria was eliminated equally successfully in both regimens. There was no significant difference between the two groups in the incidence of side effects from the drugs. The financial saving which could accrue from adopting this therapeutic regimen would be significant.