Amyloid diseases at old age : a pathological, epidemiological, and genetic study

Along with the increased life span of individuals, the burden of old age-associated diseases has inevitably increased. Alzheimer s disease (AD), probably the most well known geriatric disease, belongs to the old age-associated amyloid diseases. The purpose of this study was to investigate the frequency, genetic and health-associated risk factors, mutual association, and amyloid proteins in two old age-associated amyloid disorders senile systemic amyloidosis (SSA) and cerebral amyloid angiopathy (CAA) as part of the prospective population-based Vantaa 85+ autopsy study on a Finnish population aged 85 years or more (Studies I-III), completed with a case report on a patient with advanced AGel amyloidosis (Study IV). The numbers of patients investigated in the studies (I-III) were 256, 74, and 63, respectively. The diagnosis and grading of amyloid were based upon histological examination of tissue samples obtained post mortem and stained with Congo red. The amyloid fibril and associated proteins were characterized by immunohistochemical staining methods. The genotype frequencies of 20 polymorphisms in 9 genes and information on health-associated risk factors in subjects with and without SSA and CAA were compared. In a Finnish population ≥ 95 years of age, SSA and CAA occurred in 36% and 49% of the subjects, respectively. In total, two-thirds of these very elderly individuals had SSA, CAA, or both. However, in only 14% of the population these two conditions co-occurred. In subjects 85 years or older, the prevalence of SSA was 25%. In this population, SSA was associated with age at the time of death (p=0.002), myocardial infarctions (MIs; p=0.004), the G/G (Val/Val) genotype of the exon 24 polymorphism in the alpha2-macroglobulin (α2M) gene (p=0.042) and with the H2 haplotype of the tau gene (p=0.016). In contrast, the presence of CAA was strongly associated with APOE e4 (p=0.0003), with histopathological AD (p=0.0005), and with clinical dementia (p=0.01) in both e4+ (p=0.02) and e4- (p=0.06) individuals. Apart from demonstrating the amyloid fibril proteins, complement proteins 3d (C3d) and 9 (C9) were detected in the amyloid deposits of CAA and AGel amyloidosis, and α2M protein was found in fibrous scar tissue close to SSA. In conclusion, this first population based study on SSA shows that both SSA and CAA are common in very elderly individuals. Old age, MIs, the exon 24 polymorphism of the α2M gene, and H1/H2 polymorphism of the tau gene associate with SSA while clinical dementia and APOE ε4 genotype associate with CAA. The high prevalence of CAA, combined with its association with clinical dementia independent of APOE genotype, neuropathological AD, or SSA, also highlights its clinical significance in the very aged, among which the serious end stage complications of CAA, namely multiple infarctions and hemorrhages, are rare. The report on a patient having advanced AGel amyloidosis added knowledge on the disease and showed that this generally benign condition occasionally may lead to death. Further studies are warranted to confirm the findings in other populations. Also, the role of α2M and tau in the pathogenesis of SSA and the involvement of complement in the process of amyloid beta (Aβ) protein elimination from the brain remain to be clarified. Finally, the high prevalence of SSA in the elderly raises the need for prospective clinical studies to define its clinical significance.

Complex carbohydrates: 1. Conformational studies on some oligosaccharides related to N-glycosyl proteins which interact with concanavalin A

Empirical potential energy calculations have been carried out to determine the preferred conformations of some oligosaccharides having the trimannosidic core structure (Man3GlcNAc2) and which interact with concanavalin A. In the minimum energy conformations for the trimannosidic core the mannose residue on the Man α(1–6) arm comes close to one of the N-acetylglucosamine residues of the core. The addition of N-acetylglucosamine residues to the terminal mannose residues does not alter the preferred conformation of the trimannosidic core although it alters the relative preference of some of the higher energy conformations. The minimum energy conformation broadly agrees with available X-ray data. The presence of a bisecting N-acetylglucosamine residue on the middle mannose does not push the trimannosidic core to any new conformation but it does alter the relative preference for a particular conformation.

Role of conformational dynamics in kinetics of an enzymatic cycle in a nonequilibrium steady state

Enzyme is a dynamic entity with diverse time scales, ranging from picoseconds to seconds or even longer. Here we develop a rate theory for enzyme catalysis that includes conformational dynamics as cycling on a two-dimensional (2D) reaction free energy surface involving an intrinsic reaction coordinate (X) and an enzyme conformational coordinate (Q). The validity of Michaelis-Menten (MM) equation, i.e., substrate concentration dependence of enzymatic velocity, is examined under a nonequilibrium steady state. Under certain conditions, the classic MM equation holds but with generalized microscopic interpretations of kinetic parameters. However, under other conditions, our rate theory predicts either positive (sigmoidal-like) or negative (biphasic-like) kinetic cooperativity due to the modified effective 2D reaction pathway on X-Q surface, which can explain non-MM dependence previously observed on many monomeric enzymes that involve slow or hysteretic conformational transitions. Furthermore, we find that a slow conformational relaxation during product release could retain the enzyme in a favorable configuration, such that enzymatic turnover is dynamically accelerated at high substrate concentrations. The effect of such conformation retainment in a nonequilibrium steady state is evaluated.

Polymorphism and conformational flexibility of DNA: right and left handed duplexes

Left handed duplexes are shown to be in agreement with the X-ray intensity data of A-, B- and D-forms of DNA. The structures are stereochemically satisfactory because they were obtained following a stereochemical guideline derived from theory and single crystal structure data of nucleic acid components. The same stereochemical guideline also led to right handed duplexes for B- and D-forms of DNA which have stereochemically preferred conformation and hence are superior to those given by Arnott and coworkers.

Conformational Analysis and Electronic Structure of Monothiodiacetamide: Normal Coordinate Analysis and Molecular Orbital Study

The infrared spectra of monothiodiacetamide (MTDA, CHaCONHCSCH3) and its N-deuterated compound in solution, solid state and at low temperature are measured. Normal coordinate analysis for the planar vibrations of MTDAd o and -dl have been performed for the two most probable cis-trans-CONHCSor -CSNHCO-conformers using a simple Urey-Bradley force function. The conformation of MTDA derived from the vibrational spectra is supported by the all valence CNDO/2 molecular orbital method. The vibrational assignments and the electronic structure of MTDA are also given.

Theoretical studies on Beta-lactam antibiotics VI: Conformational analysis and structure-activity relationships of penicillin sulfoxides and cephalosporins

Conformational energy calculations were carried out on penicillin α-and Β-sulfoxides and δ2- and δ3-cephalosporins, in order to identify the structural features governing their biological activity. Results on penicillin Β-sulfoxide indicated that in its favoured conformation, the orientation of the aminoacyl group was different from the one required for biological activity. Penicillin α sulfoxide, like penicillin sulfide, favoured two conformations of nearly equal energies, but separated by a much higher energy barrier. The reduced activity of the sulfoxides despite the nonplanarity of their lactam peptide indicated that the orientations of the aminoacyl and carboxyl groups might also govern biological activity. δ3-cephalosporins favoured two conformations of nearly equal energies, whereas δ2-cephalosporins favoured only one conformation. The lactam peptide was moderately nonplanÄr in the former, but nearly planar in the latter. The differences in the.preferred orientations of the carboxyl group between penicillins and cephalosporins were correlated with the resistance of cephalosporins to penicillinases.

Evaluation of the use of prochloraz in the control of postharvest diseases of papaya in Australia

Prochloraz as Sportak at 450 g a.i./L is registered for the control of postharvest diseases in papaya in Australia. A project in far north Queensland in 2011, examined the use patterns of postharvest treatments, evaluated treatment dips and sprays for prochloraz concentrations and evaluated the efficacy of prochloraz at 0, 20, 40, 55 and 70 ml/100 L, fludioxonil as Scholar at 260 ml/100 L and azoxystrobin as Amistar at 50 ml/100 L. Results showed that packing shed use of Sportak varied with recycled and stored solutions showing a depletion of the active ingredient. Measured prochloraz in solution was highly pH dependent with nominal solution values only being measured when the pH was less than 3.0. In the fungicide efficacy trial Sportak at the label rate of 55 ml/100 L provided more effective disease control than fludioxonil and azoxystrobin. The trial also suggested that fruit from older trees showed a high degree of disease incidence relative to fruit from young trees.

Timing of Infection and Development of Alternaria Diseases in the Canopy of Apple Trees

Alternaria leaf blotch and fruit spot of apple caused by Alternaria spp. cause annual losses to the Australian apple industry. Erratic control using protectant fungicides is often experienced and may be due to the lack of understanding of the timing of infection and epidemiology of the diseases. We found that Alternaria leaf blotch infection began about 20 days after bloom (DAB) and the highest disease incidence occurred from 70 to 110 DAB. Alternaria fruit spot infection occurred about 100 DAB in the orchard. Fruit inoculations in planta showed that there was no specific susceptible stage of fruit. Leaves and fruit in the lower canopy of trees showed higher levels of leaf blotch and fruit spot incidence than those in the upper canopy and the incidence of leaf blotch in shoot leaves was higher than in spur leaves. Temperature, relative humidity, and rainfall affected leaf blotch and fruit spot incidence. The gained knowledge on the timing of infection and development of disease may aid in the development of more effective disease management strategies.

Conformational studies of phynyl and isoxazolyl penicillins

Conformational energy calculations have been carried out on a few representative b-lactamase resistant and susceptible, phenyl and isoxazolyl penicillins. These studies, in agreement with those of earlier workers, show that the 6b-side chains of resistants penicillins are highly rigid as compared to those of susceptible penicillins. The present studies also suggest that the degree of resistant to b-lactamases depends not only on the rigidity of the side chain but also on the nature and orientation of the substituent, beyond the amide carbonyl group in the side chain. The overall shapes of these penicillins correlate well with their antibacterial properties.

The use of plant activators in mango postharvest diseases management

Anthracnose and stem end rots are the main postharvest diseases affecting mangoes in Australia and limiting the shelf life of fruits whenever they are not controlled. The management of these diseases has often relied on the use of fungicide applications either as field spray treatments, postharvest dips or both. Because of concerns with continuous fungicide use, other options for the sustainable management of these diseases are needed. Field trials were conducted to assess the efficacy of three plant activators for the control of these diseases over a 2-year period on 20-year old ‘R2E2’ mango trees in north Queensland. The activators evaluated were: Bion, Kasil and Mangocote. The efficacy of these activators was compared with that of a standard industry field spray program using a combination of fungicides, as well as to un¬treated controls. Conditions favoured good development of the target diseases in both years to be able to differentiate treatment effects. Kasil as a drench was as effective as the standard fungicide program on the management of anthracnose and stem end rots. Bion as foliar sprays showed similar efficacy with its effectiveness comparable with the standard spray program. Both activators had significantly less disease incidences when compared with the untreated control. The third activator, Mangocote was not very effective in controlling the target diseases. Its effect was not significantly better than the untreated controls. The results from this 2-year study suggest that plant activators can play an effective role in mango postharvest disease management. Proper timing could reduce the number of fungicide sprays in an integrated disease management program enabling sustainable yields of quality fruits without the continuous concerns of health and environmental risks from continuous reliance on fungicide use.

Comparison of fungicide dips efficacies for the control of mango postharvest diseases

The shelf-life of mangoes is limited by two main postharvest diseases when not consistently managed. These are anthracnose ( Colletotrichum gloeosporioides) and stem end rots ( Neofusicoccum parvum). The management of these diseases has often relied mainly on the use of fungicide applications either as field spray treatments and/or postharvest dips. Current postharvest dips are under continuous threats because of health concerns and the maximum residue levels allowed on treated fruit continuous to be reviewed and re-assessed. Research needs to keep up with the rate at which changes are occurring following some of these reviews. The recent withdrawal of carbendazin (Spinflo), as a postharvest dip being used to manage stem end rots necessitated the urgent search for a replacement fungicide to manage this disease. A study was therefore undertaken to compare the efficacy of current and potential products that could be used to fill the gap. The following products were evaluated: Carbendazin (Spinflo), Prochloraz (Sportak), Thiobendazole (TBZ) and Fludioxonil (Scholar). These products were tested both under ambient temperatures and as hot dips to identify one that was most effective. Scholar as a hot dip was the most effective product among the ones compared. It effectively controlled both anthracnose and stem end rots at highly significant levels when compared to the untreated control and even Spinflo which is being replaced. As a cold dip, it had some limited effect on anthracnose but had virtually no effect on stem end rots. Based on its performance in these experiments, the product has been recommended for rates and residue studies so that it can be registered as a hot dip for use in controlling postharvest diseases of mangoes.

Networks of stored grain diseases and pests: Strategies for sampling and mitigation

Pathogens and pests of stored grains move through complex dynamic networks linking fields, farms, and bulk storage facilities. Human transport and other forms of dispersal link the components of this network. A network model for pathogen and pest movement through stored grain systems is a first step toward new sampling and mitigation strategies that utilize information about the network structure. An understanding of network structure can be applied to identifying the key network components for pathogen or pest movement through the system. For example, it may be useful to identify a network node, such as a local grain storage facility, through which grain from a large number of fields will be accumulated and move through the network. This node may be particularly important for sampling and mitigation. In some cases more detailed information about network structure can identify key nodes that link two large sections of the network, such that management at the key nodes will greatly reduce the risk of spread between the two sections. In addition to the spread of particular species of pathogens and pests, we also evaluate the spread of problematic subpopulations, such as subpopulations with pesticide resistance. We present an analysis of stored grain pathogen and pest networks for Australia and the United States.

Foliar, fruit and soilborne diseases.

This chapter provides an in-depth review of important diseases affecting avocado production throughout the world. The importance of understanding the interaction of plant pathogens with their avocado host in order for the development of disease management options is also discussed.