Structure-activity relationship of amyloid fibrils

Protein aggregation is a process in which proteins self-associate into imperfectly ordered macroscopic entities. Such aggregates are generally classified as either amorphous or highly ordered, the most common form of the latter being amyloid fibrils. Amyloid fibrils composed of cross-β-sheet structure are the pathological hallmarks of several diseases including Alzheimer’s disease, but are also associated with functional states such as the fungal HET-s prion. This review aims to summarize the recent high-resolution structural studies of amyloid fibrils in light of their (potential) activities. We propose that the repetitive nature of the cross-β-sheet structure of amyloids is key for their multiple properties: the repeating motifs can translate a rather non-specific interaction into a specific one through cooperativity.     

Mistic: Cellular localization,solution behavior,polymerization, and fibril formation

Mistic represents a family of unique membrane‐associating proteins originally found in Bacillus subtilis (M110). As a fusion partner, it has been shown to assist overexpression of foreign integral membrane proteins in E. coli. We have expressed shorter Mistic homologs from other Bacillus species and surprisingly, unlike M110, found them abundant in the cytoplasm. These Mistic homologs including the corresponding shorter sequence (amino acids 27 through 110 of M110) exist as multimeric assemblies in solution in the absence of detergent. Crystals of Mistic from B. leicheniformis (M2) diffracted to 3.2 Å resolution, indicating that it exists as a multimer in the crystalline state as well. Moreover, we show that although M2 is mostly α‐helical, it tends to polymerize and form fibrils. Such oligomerization could potentially mask the charged surface of the monomeric Mistic to assist membrane integration.     

Home ranges, movements, and activity of wolves (Canis lupus) in the Dalmatian part of Dinarids, Croatia

Home-range sizes, movements, and daily activity of wolves (Canis lupus L. 1758) were studied in Dalmatia, Croatia in 1998–2001. The total home ranges (100% MCP) of two packs were 160 km² and 141 km², mean=150.5 km². Core areas (50% kernel) were 26.2 km² and 3.3 km², respectively. Differences in core area sizes were influenced by human activity—hunting and sheep grazing. Compared with random locations, wolf locations were closer to the nearest water source (mean=937 m) and farther from houses (mean=653 m). Wolves were significantly more active during the night than during the day (activity indexes were 0.53 vs. 0.35), and night activity was higher during summer (0.58), and lower during winter (0.48). A correlation was found between distances traveled and activity index (r=0.58, p=0.003). Home range, seasonal variations in home-range size, habitat use, and activity of wolves in Dalmatia were oriented to make the compromise from danger of proximity to humans and also to benefit from human-related food sources.     

Amyloid as a depot for the formulation of long-acting drugs

Amyloids are highly organized protein aggregates that are associated with both neurodegenerative diseases such as Alzheimer disease and benign functions like skin pigmentation. Amyloids self-polymerize in a nucleation-dependent manner by recruiting their soluble protein/peptide counterpart and are stable against harsh physical, chemical, and biochemical conditions. These extraordinary properties make amyloids attractive for applications in nanotechnology. Here, we suggest the use of amyloids in the formulation of long-acting drugs. It is our rationale that amyloids have the properties required of a long-acting drug because they are stable depots that guarantee a controlled release of the active peptide drug from the amyloid termini. This concept is tested with a family of short- and long-acting analogs of gonadotropin-releasing hormone (GnRH), and it is shown that amyloids thereof can act as a source for the sustained release of biologically active peptides.     

Naturally Occurring Variants of the Dysglycemic Peptide Pancreastatin: DIFFERENTIAL POTENCIES FOR MULTIPLE CELLULAR FUNCTIONS AND STRUCTURE-FUNCTION CORRELATION*

Pancreastatin (PST), a chromogranin A-derived peptide, is a potent physiological inhibitor of glucose-induced insulin secretion. PST also triggers glycogenolysis in liver and reduces glucose uptake in adipocytes and hepatocytes. Here, we probed for genetic variations in PST sequence and identified two variants within its functionally important carboxyl terminus domain: E287K and G297S. To understand functional implications of these amino acid substitutions, we tested the effects of wild-type (PST-WT), PST-287K, and PST-297S peptides on various cellular processes/events. The rank order of efficacy to inhibit insulin-stimulated glucose uptake was: PST-297S > PST-287K > PST-WT. The PST peptides also displayed the same order of efficacy for enhancing intracellular nitric oxide and Ca²⁺ levels in various cell types. In addition, PST peptides activated gluconeogenic genes in the following order: PST-297S ≈ PST-287K > PST-WT. Consistent with these in vitro results, the common PST variant allele Ser-297 was associated with significantly higher (by ∼17 mg/dl, as compared with the wild-type Gly-297 allele) plasma glucose level in our study population (n = 410). Molecular modeling and molecular dynamics simulations predicted the following rank order of α-helical content: PST-297S > PST-287K > PST-WT. Corroboratively, circular dichroism analysis of PST peptides revealed significant differences in global structures (e.g. the order of propensity to form α-helix was: PST-297S ≈ PST-287K > PST-WT). This study provides a molecular basis for enhanced potencies/efficacies of human PST variants (likely to occur in ∼300 million people worldwide) and has quantitative implications for inter-individual variations in glucose/insulin homeostasis.     

Genetic variability in mulberry against Myrothecium leaf spot (Myrothecium roridum) and identification of resistance genotypes

Disease response of 30 mulberry genotypes to Myrothecium leaf spot (Myrothecium roridum) was studied under inoculated condition. It was observed that 10 genotypes were resistant, 16 genotypes moderately resistant and 4 genotypes moderately susceptible to the disease. Area under disease progress curve (AUDPC) and apparent infection rate was found significantly lower in the resistant genotypes. Correlation study revealed that percent disease index (PDI) has significant positive correlation with AUDPC and apparent infection rate. Genetic analysis of disease-resistant traits (PDI and mean AUDPC) revealed that phenotypic coefficient of variation (PCV) was higher than genotypic coefficient of variation (GCV) for both PDI and AUDPC, and GCV/PCV ratio was also found high which indicated that disease-resistant traits to Myrothecium leaf spot were not much influenced by environment. High heritability coupled with high genetic advance indicated that the disease-resistant traits are due to additive gene effect thereby indicating the amenability of disease-resistant in the selection process. Hence disease-resistant mulberry genotypes viz. C-763, S-34, Jodhpur, Cyprus, Australian and Hungarian may be used as source of resistance to Myrothecium leaf spot for future breeding programme. Besides, high yielding genotypes viz. Tr-10, C-763 and S-34 may be recommended for commercial exploitation     

Aging exacerbates damage and delays repair of alveolar epithelia following influenza viral pneumonia

Background

Influenza virus infection causes significantly higher levels of morbidity and mortality in the elderly. Studies have shown that impaired immunity in the elderly contributes to the increased susceptibility to influenza virus infection, however, how aging affects the lung tissue damage and repair has not been completely elucidated.

Methods

Aged (16–18 months old) and young (2–3 months old) mice were infected with influenza virus intratracheally. Body weight and mortality were monitored. Different days after infection, lung sections were stained to estimate the overall lung tissue damage and for club cells, pro-SPC⁺ bronchiolar epithelial cells, alveolar type I and II cells to quantify their frequencies using automated image analysis algorithms.

Results

Following influenza infection, aged mice lose more weight and die from otherwise sub-lethal influenza infection in young mice. Although there is no difference in damage and regeneration of club cells between the young and the aged mice, damage to alveolar type I and II cells (AT1s and AT2s) is exacerbated, and regeneration of AT2s and their precursors (pro-SPC-positive bronchiolar epithelial cells) is significantly delayed in the aged mice. We further show that oseltamivir treatment reduces virus load and lung damage, and promotes pulmonary recovery from infection in the aged mice.

Conclusions

These findings show that aging increases susceptibility of the distal lung epithelium to influenza infection and delays the emergence of pro-SPC positive progenitor cells during the repair process. Our findings also shed light on possible approaches to enhance the clinical management of severe influenza pneumonia in the elderly.

Electronic supplementary material

The online version of this article (doi:10.1186/s12931-014-0116-z) contains supplementary material, which is available to authorized users.     

Tono Reservoir fishery contribution to poverty reduction among fishers in northern Ghana

Fishery characteristics and livelihood status of fishers at Tono Reservoir, Ghana, were investigated between January 2015 and June 2016. Data on fisher demography, fishing gears, fishing methods, perceptions of the state of fish stocks, management practices, income and consumption of fishers were obtained through structured interviews. Censuses of fishers and fishing gears were conducted through direct observation and counts. The population of fishers was 950 and the majority (74%) of the sampled respondents fell within the ages of 24–41 years. Gillnet, cast net, trap and hook and line were the four main gears utilised. Illegal methods of fishing observed included the use of mosquito nets (nets with mesh <1.0 cm) and the use of brewer’s waste (pito mash) as bait. Brycinus nurse, Synodontis spp., Parailia spiniserrata and Chrysichthys spp. were perceived to have disappeared from the reservoir. The fishers were unaware of the existence of any fisheries regulations, hence there was no adherence to management practices. Their daily income was derived mainly from fishing. The incidence of poverty among fishers was low (8%). The Tono Reservoir has a great potential for supporting livelihood if it is properly managed.     

臭虫击倒抗性基因突变检测及抗性测定

近20多年,臭虫（Cimex spp.）在世界范围内成为常见的卫生害虫,其防治主要采用化学防治,但很多种群发现击倒抗性（Knockdown resistance gene, kdr）基因突变的存在以及抗药性。监测kdr的发生频率以及不同种群对农药的抗性对臭虫有效防治很重要,但我国对臭虫种群的抗药性报道很少。本试验采用点滴法测定了1个温带臭虫Cimex lectularius野外种群对氯虫苯甲酰胺、呋虫胺、吡虫啉、噻虫嗪和高效氯氰菊酯等5种药剂的毒性及抗性水平,使用区分剂量快速鉴定抗性方法对2个温带臭虫和2个热带臭虫Cimex hemipterus种群对高效氯氰菊酯的抗性水平进行了检测,此外用PCR方法检测8个臭虫地理种群（1个温带臭虫实验室种群,1个温带臭虫野外种群和6个热带臭虫野外种群）174个个体的kdr突变频率。点滴法结果表明,5种杀虫剂对温带臭虫的毒性是吡虫啉和呋虫胺>噻虫嗪>氯虫苯甲酰胺和高效氯氰菊酯,测试的野外温带臭虫种群仅对噻虫嗪无明显抗性。热带臭虫2个野外种群对高效氯氰菊酯的抗性均远高于温带臭虫。在温带臭虫的实验室种群中未检测到突变,在野外种群中检测到了V419L和L925I突变,可分为2种基因型类型（A：无突变位点;B：同时有L925I和V419L）,而在热带臭虫的6个种群检测到M918I和L1014F突变,只有1种基因型类型,即M918I和L1014F双位点突变。温带臭虫1个野外种群及热带臭虫6个野外种群kdr突变的存在与臭虫对高效氯氰菊酯敏感密切关联。基于kdr基因突变检测结果推测我国臭虫种群广泛存在对拟除虫菊酯的抗性。     

Cell Adhesion on Amyloid Fibrils Lacking Integrin Recognition Motif

Amyloids are highly ordered, cross-β-sheet-rich protein/peptide aggregates associated with both human diseases and native functions. Given the well established ability of amyloids in interacting with cell membranes, we hypothesize that amyloids can serve as universal cell-adhesive substrates. Here, we show that, similar to the extracellular matrix protein collagen, amyloids of various proteins/peptides support attachment and spreading of cells via robust stimulation of integrin expression and formation of integrin-based focal adhesions. Additionally, amyloid fibrils are also capable of immobilizing non-adherent red blood cells through charge-based interactions. Together, our results indicate that both active and passive mechanisms contribute to adhesion on amyloid fibrils. The present data may delineate the functional aspect of cell adhesion on amyloids by various organisms and its involvement in human diseases. Our results also raise the exciting possibility that cell adhesivity might be a generic property of amyloids.