Surface-Induced Hydrophobin Assemblies with Versatile Properties and Distinct Underlying Structures.

Hydrophobins are remarkable proteins due to their ability to self-assemble into amphipathic coatings that reverse surface wettability. Here, the versatility of the Class I hydrophobins EASΔ15 and DewY in diverse nanosuspension and coating applications is demonstrated. The hydrophobins are shown to coat or emulsify a range of substrates including oil, hydrophobic drugs, and nanodiamonds and alter their solution and surface behavior. Surprisingly, while the coatings confer new properties, only a subset is found to be resistant to hot detergent treatment, a feature previously thought to be characteristic of the functional amyloid form of Class I hydrophobins. These results demonstrate that substrate surface properties can influence the molecular structures and physiochemical properties of hydrophobin and possibly other functional amyloids. Functional amyloid assembly with different substrates and conditions may be analogous to the propagation of different polymorphs of disease-associated amyloid fibrils with distinct structures, stability, and clinical phenotypes. Given that amyloid formation is not required for Class I hydrophobins to serve diverse applications, our findings open up new opportunities for their use in applications requiring a range of chemical and physical properties. In hydrophobin nanotechnological applications where high stability of assemblies is required, simultaneous structural and functional characterization should be carried out. Finally, while results in this study pertain to synthetic substrates, they raise the possibility that at least some members of the pseudo-Class I and Class III hydrophobins, reported to form assemblies with noncanonical properties, may be Class I hydrophobins adopting alternative structures in response to environmental cues.

Perphenazine-Macrocycle Conjugates Rapidly Sequester the Aß42 Monomer and Prevent Formation of Toxic Oligomers and Amyloid.

Alzheimer's disease is imposing a growing social and economic burden worldwide, and effective therapies are urgently required. One possible approach to modulation of the disease outcome is to use small molecules to limit the conversion of monomeric amyloid (Aß42) to cytotoxic amyloid oligomers and fibrils. We have synthesized modulators of amyloid assembly that are unlike others studied to date: these compounds act primarily by sequestering the Aß42 monomer. We provide kinetic and nuclear magnetic resonance data showing that these perphenazine conjugates divert the Aß42 monomer into amorphous aggregates that are not cytotoxic. Rapid monomer sequestration by the compounds reduces fibril assembly, even in the presence of pre-formed fibrillar seeds. The compounds are therefore also able to disrupt monomer-dependent secondary nucleation, the autocatalytic process that generates the majority of toxic oligomers. The inhibitors have a modular design that is easily varied, aiding future exploration and use of these tools to probe the impact of distinct Aß42 species populated during amyloid assembly.

Carbon/iron by-product from catalytic methane decomposition as recyclable Fenton catalyst for pollutant degradation.

Direct catalytic decomposition of methane (CDM) has been studied as a possible emission-free hydrogen production route for over 100 years. However, the high cost of catalyst regeneration limits its practical applications. Here, we demonstrate that the solid by-product from CDM using Fe ore catalysts comprising carbon nano onions encapsulated with magnetic Fe cores (Fe@C) can serve as efficient and recyclable Fenton catalysts for pollutant degradation. Fe@C/H2O2 has better performance than FeSO4/H2O2 at similar Fe concentrations and can be used to decompose various pollutants. Mechanistic studies reveal that graphitic carbon layers and encapsulated Fe0 contribute to their high catalytic activity. Further, Fe@C can be easily recovered from an aqueous solution and reused due to the encapsulated magnetic Fe particles. Over three reused cycles, Fe@C/H2O2 only yields 1/8 of Fe sludges compared to FeSO4/H2O2, significantly reducing Fe sludge treatment costs. Overall, Fe@C demonstrates excellent application potentials in water and wastewater treatment, making H2 production via CDM economically more viable.

The impact of Timothy's Law on hospitalization among patients with mental health conditions in New York State.

BACKGROUND: Timothy's law to reduce mental health care disparities was enacted in January 2007 in New York state (NY). According to Timothy's law, "if a patient is suffering from a Biologically Based Mental Illness, or is a Child with Serious Emotional Disturbances, the Inpatient mental health benefit will be the same as for any other illness". An assessment of its impact on inpatient mental health care is lacking. We provide a rigorous study of this policy intervention's effect over the first year of its implementation. METHODS: We used a quasi-experimental design to combine the difference-in-difference method and propensity score weighting. Data are from inpatient records in NY and California (CA) (as a control) between January 2006 to December 2006 (the pre-enactment year in NY) and January to December 2007 (the enactment year) for non-Medicare/Medicaid patients hospitalized in both years with specific illnesses covered by Timothy's Law. Change in length of stay from 2006 to 2007 was measured for each patient, and the differences observed in NY and California were compared to each other (Difference-in-Difference), with differences in the characteristics of patients in NY and California addressed through Propensity Score Weighting (PSW). RESULTS: Before Timothy's Law was enacted (2006), length of stay (LOS) in NY was 16.3 days on average, and length of stay per hospitalization (LOSPH) was 11.72 days on average for the 1237 patients under study in 2006. In 2007, LOS increased by 4.91 days in NY (95% CI (2.89, 7.01)) compared with similar patients in California, and LOSPH by 3.25 days (95% CI (1.96, 4.57)). Among patients with serious mental illness diagnoses, LOS in NY increased by 7.07 days (95% CI (4.15, 10.17)), and LOSPH by 4.04 days (95% CI (1.93, 6.03)) compared to California. CONCLUSIONS: Our study strongly suggests that, within the time frame of just a single year, Timothy's Law significantly increased inpatient mental healthcare utilization in NY. Our study raises the possibility that similar laws in other locations could have similar effects.

Can We Use Grip Strength to Predict Other Types of Hand Exertions? An Example of Manufacturing Industry Workers.

BACKGROUND: There are different types of hand motions in people's daily lives and working environments. However, testing duration increases as the types of hand motions increase to build a normative database. Long testing duration decreases the motivation of study participants. The purpose of this study is to propose models to predict pinch and press strength using grip strength. METHODS: One hundred ninety-eight healthy volunteers were recruited from the manufacturing industries in Central Taiwan. The five types of hand motions were grip, lateral pinch, palmar pinch, thumb press, and ball of thumb press. Stepwise multiple linear regression was used to explore the relationship between force type, gender, height, weight, age, and muscle strength. RESULTS: The prediction models developed according to the variable of the strength of the opposite hand are good for explaining variance (76.9-93.1%). Gender is the key demographic variable in the predicting models. Grip strength is not a good predictor of palmar pinch (adjusted-R2: 0.572-0.609), nor of thumb press and ball of thumb (adjusted-R2: 0.279-0.443). CONCLUSIONS: We recommend measuring the palmar pinch and ball of thumb strength and using them to predict the other two hand motions for convenience and time saving.

Body volume estimation equation for male laborers in Taiwan.

The purpose of this study was to develop a body volume (BV) estimation equation for male laborers in Taiwan with body weight (W) and stature height (H) as initial estimators. A three-dimensional (3D) body scanner and a 3D foot scanner were used to measure the 3D range data of 100 male laborers in this study. Subjects' BV was extracted from the 3D body model, and H and W were used as independent variables in regression analysis. The results show that the final BV estimation equation is BV = 1122.927 × W0.972, with R2 = 0.949. Thirty extra male subjects were scanned to compare this BV estimation equation with those in previous studies. The results show that this BV estimation equation had the smallest absolute mean difference at 1.1458 L and the smallest standard error of the estimate at 2.48% in comparison.

Investigation into the Effects of Backrest Angle and Stick Location on Female Strength.

OBJECTIVES: The purpose of this study was to investigate the effects of backrest angle and hand maneuver direction on maximum hand strength and to recommend a strength value for the hand-controlled stick of an aircraft. METHODS: Forty-eight female subjects were recruited to perform simulated forward-backward and adduction-abduction maneuvers using control sticks. Each subject was free from musculoskeletal disorders and pain. The independent variables included four control maneuvers (forward, backward, adduction, abduction), two right-hand control stick locations (central, side), and three backrest angles (90°, 103°, 108°). The dependent variable was maximum hand strength. RESULTS: The maximum strength for forward maneuvers with both central and side sticks was strongest at a 90° backrest angle (p < 0.001). The maximum strength for adduction maneuvers with both central and side sticks was also strongest at a 90° backrest angle (p < 0.001). On the other hand, the highest strength was observed at a 108° backrest angle when pulling the stick backward (p < 0.001). The abduction strength was significantly stronger than the adduction strength with a central stick (p < 0.001), but the adduction strength was significantly stronger than the abduction strength with a side stick (p < 0.001-p = 0.017). The forward and abduction strength were significantly different in different locations (p < 0.001). The recommended strength in the Code of Federal Regulations (CFR) by the US FAA is higher than the strength values observed in this study. CONCLUSIONS: The backrest angle, directions, and location affected the muscular strength. The recommended values should be reevaluated and adjusted for Taiwanese pilots.

Management of Patients With Sepsis in Canadian Community Emergency Departments: A Retrospective Multicenter Observational Study.

BACKGROUND: Sepsis is a life-threatening syndrome and a leading cause of morbidity and mortality representing significant financial burden on the health-care system. Early identification and intervention is crucial to maximizing positive outcomes. We studied a quality improvement initiative with the aim of reviewing the initial management of patients with sepsis in Canadian community emergency departments, to identify areas for improving the delivery of sepsis care. We present a retrospective, multicenter, observational study during 2011 to 2015 in the community setting. METHODS: We collected data on baseline characteristics, clinical management metrics (triage-to-physician-assessment time, triage-to-lactate-drawn time, triage-to-antibiotic time, and volume of fluids administered within the first 6 hours of triage), and outcomes (intensive care unit [ICU] admission, in-hospital mortality) from a regional database. RESULTS: A total of 2056 patients were analyzed. The median triage-to-physician-assessment time was 50 minutes (interquartile range [IQR]: 25-104), triage-to-lactate-drawn time was 50 minutes (IQR: 63-94), and triage-to-antibiotics time was 129 minutes (IQR: 70-221). The median total amount of fluid administered within 6 hours of triage was 2.0 L (IQR: 1.5-3.0). The ICU admission rate was 36% and in-hospital mortality was 25%. We also observed a higher ICU admission rate (51% vs 24%) and in-hospital mortality (44% vs 14%) in those with higher lactate concentration (≥4 vs ≤2 mmol/L), independent of other sepsis-related parameters. CONCLUSION: Time-to-physician-assessment, time-to-lactate-drawn, time-to-antibiotics, and fluid resuscitation in community emergency departments could be improved. Future quality improvement interventions are required to optimize management of patients with sepsis. Elevated lactate concentration was also independently associated with ICU admission rate and in-hospital mortality rate.

Normative Hand Strength of Healthcare Industry Workers in Central Taiwan.

Objectives: The purpose of this study is to establish the norms of hand grip strength in the healthcare industry in Taiwan and propose models to predict the strength of hand movement by regression with demographic and anthropometric factors. Methods: This is a cross-sectional study with a stratified convenience sample of workers in healthcare service industries in central Taiwan. Three hundred twenty-nine healthy subjects were recruited. Strength of different hand movement were tested three times in both hands and rests were given between tests. Results: Female strength of these hand movement was 59.1% to 73.0% that in males (p < 0.001). In general, the hand strength of male workers in the healthcare industry was less than that of male workers in the manufacturing industry (p < 0.001). In the prediction model, sex and weight played important roles in predicting hand strength. Conclusions: The norms of different types of hand strength was investigated the first time in workers in the healthcare industry in Taiwan. The tasks performed by healthcare personnel vary widely, and this variable should be considered in a future prediction model.

Formation of Amphipathic Amyloid Monolayers from Fungal Hydrophobin Proteins.

The fungal hydrophobins are small proteins that are able to self-assemble spontaneously into amphipathic monolayers at hydrophobic:hydrophilic interfaces. These protein monolayers can reverse the wettability of a surface, making them suitable for increasing the biocompatibility of many hydrophobic nanomaterials. One subgroup of this family, the class I hydrophobins, forms monolayers that are composed of extremely robust amyloid-like fibrils, called rodlets. Here, we describe the protocols for the production and purification of recombinant hydrophobins and oxidative refolding to a biologically active, soluble, monomeric form. We describe methods to trigger the self-assembly into the fibrillar rodlet state and techniques to characterize the physicochemical properties of the polymeric forms.

A Pilot Study of Five Types of Maximum Hand Strength among Manufacturing Industry Workers in Taiwan.

Background: The purpose of this study is to collect five types of maximum hand strength among workers in the manufacturing industry in Taiwan. Methods: This study is a cross sectional study with a stratified and convenient sample of workers on the production line in manufacturing industries in Central Taiwan. In total, we recruited 198 healthy subjects to participate in this study. Five types of hand strength were measured in both hands three times with 3 min rests between trials. Results: The strength of females for these five types of hand exertions were 52.0% to 67.6% of the strength of males (p < 0.001). For both genders, there was a main effect for the types of hand strength for the right hand (p < 0.001) and the left hand (p < 0.001). In general, the hand strength in U.S. and EU countries was 1.2 to 1.7 times greater than the strength among the three types of hand exertions in this study. Conclusion: These results can be used to evaluate the musculoskeletal burdens on the upper extremities in the manufacturing industry and could also be used for tool and job design and job modifications.

Fungal Hydrophobins and Their Self-Assembly into Functional Nanomaterials.

In recent years, much attention has focused on incorporating biological and bio-inspired nanomaterials into various applications that range from functionalising surfaces and enhancing biomolecule binding properties, to coating drugs for improved bioavailability and delivery. Hydrophobin proteins, which can spontaneously assemble into amphipathic layers at hydrophobic:hydrophilic interfaces, are exciting candidates for use as nanomaterials. These unique proteins, which are only expressed by filamentous fungi, have been the focus of increasing interest from the biotechnology industry, as evidenced by the sharply growing number of hydrophobin-associated publications and patents. Here, we explore the contribution of different hydrophobins to supporting fungal growth and development. We describe the key structural elements of hydrophobins and the molecular characteristics that underlie self-assembly of these proteins at interfaces. We outline the multiple roles that hydrophobins can play in supporting aerial growth of filamentous structures, facilitating spore dispersal and preventing an immune response in the infected host. The growing understanding of the hydrophobin protein structure and self-assembly process highlights the potential for hydrophobin proteins to be engineered for use in a variety of novel applications that require biocompatible coatings.

An overview of technologies to recover phosphorus as struvite from wastewater: advantages and shortcomings.

Phosphorus (P) is a significant limiting nutrient which is essential for all forms of lives. However, phosphate rock reserves are depleting rapidly due to population growth. At the same time, several countries have imposed legislative regulations on P-release into surface waters due to eutrophication. Nutrient recovery from wastewater can facilitate a sustainable, cost-effective and environment-friendly source of phosphorus. Although P-recovery as struvite from wastewater has been widely studied for a long time, there still exists a lot of challenges for widespread full-scale implementation. This paper presents a comprehensive analysis of the current state of the technologies for phosphorus recovery in the form of struvite. Fluidized bed reactors (FBRs) are widely used compared to continuously stirred reactors for P-recovery as struvite because of different solid and liquid retention time. Commercially available technologies were reported to accomplish about 80% P-removal efficiencies with a reasonable P-recovery for the most of the cases. The struvite production rate of various technologies varies from 0.89 to 13.7 kg/kg influent P. Nevertheless, these technologies are associated with several shortcomings such as high operational costs, high energy consumption, and large footprint. Increasing efforts focusing on the development of sustainable and commercially feasible technologies are expected in this sector as P-recovery is considered to be the future of wastewater engineering.

Impact of supersaturation ratio on phosphorus recovery from synthetic anaerobic digester supernatant through a struvite crystallization fluidized bed reactor.

Over the past few decades, several technologies have been developed to recover phosphorus (P) as struvite from wastewater. Although these technologies have achieved reasonable P-removal efficiencies, these technologies are associated with several shortcomings such as high capital and operating costs, longer crystallization time and production of low-quality product. This study focussed on the development of an efficient technology by designing a new fluidized bed reactor (FBR) and determining its optimum operating conditions. The supersaturation ratio is the most important process parameter for struvite recovery. This study exerted effort to establish a range of supersaturation ratios in order to achieve optimum P-removal and recovery with a lesser amount of fine crystals produced. Bench-scale FBR used in this study was able to accomplish 90% P-removal with 18% P-recovery. P-removal efficiency was observed to be increasing with an increase in the initial supersaturation ratio up to a value of 6.5. On the other hand, an increase in the supersaturation ratio resulted in a lower P-recovery efficiency with an increase in fines production. The supersaturation ratio from 5.5 to 6.0 was found to be optimum for efficient operation of the reactor.

Assembly and disassembly of Aspergillus fumigatus conidial rodlets.

The rodlet structure present on the Aspergillus fumigatus conidial surface hides conidia from immune recognition. In spite of the essential biological role of the rodlets, the molecular basis for their self-assembly and disaggregation is not known. Analysis of the soluble forms of conidia-extracted and recombinant RodA by NMR spectroscopy has indicated the importance of disulfide bonds and identified two dynamic regions as likely candidates for conformational change and intermolecular interactions during conversion of RodA into the amyloid rodlet structure. Point mutations introduced into the RODA sequence confirmed that (1) mutation of a single cysteine was sufficient to block rodlet formation on the conidial surface and (2) both presumed amyloidogenic regions were needed for proper rodlet assembly. Mutations in the two putative amyloidogenic regions retarded and disturbed, but did not completely inhibit, the formation of the rodlets in vitro and on the conidial surface. Even in a disturbed form, the presence of rodlets on the surface of the conidia was sufficient to immunosilence the conidium. However, in contrast to the parental conidia, long exposure of mutant conidia lacking disulfide bridges within RodA or expressing RodA carrying the double (I115S/I146G) mutation activated dendritic cells with the subsequent secretion of proinflammatory cytokines. The immune reactivity of the RodA mutant conidia was not due to a modification in the RodA structure, but to the exposure of different pathogen-associated molecular patterns on the surface as a result of the modification of the rodlet surface layer. The full degradation of the rodlet layer, which occurs during early germination, is due to a complex array of cell wall bound proteases. As reported earlier, this loss of the rodlet layer lead to a strong anti-fumigatus host immune response in mouse lungs.

Probing Structural Changes during Self-assembly of Surface-Active Hydrophobin Proteins that Form Functional Amyloids in Fungi.

Hydrophobins are amphiphilic proteins secreted by filamentous fungi in a soluble form, which can self-assemble at hydrophilic/hydrophobic or water/air interfaces to form amphiphilic layers that have multiple biological roles. We have investigated the conformational changes that occur upon self-assembly of six hydrophobins that form functional amyloid fibrils with a rodlet morphology. These hydrophobins are present in the cell wall of spores from different fungal species. From available structures and NMR chemical shifts, we established the secondary structures of the monomeric forms of these proteins and monitored their conformational changes upon amyloid rodlet formation or thermal transitions using synchrotron radiation circular dichroism and Fourier-transform infrared spectroscopy (FT-IR). Thermal transitions were followed by synchrotron radiation circular dichroism in quartz cells that allowed for microbubbles and hence water/air interfaces to form and showed irreversible conformations that differed from the rodlet state for most of the proteins. In contrast, thermal transitions on hermetic calcium fluoride cells showed reversible conformational changes. Heating hydrophobin solutions with a water/air interface on a silicon crystal surface in FT-IR experiments resulted in a gain in ß-sheet content typical of amyloid fibrils for all except one protein. Rodlet formation was further confirmed by electron microscopy. FT-IR spectra of pre-formed hydrophobin rodlet preparations also showed a gain in ß-sheet characteristic of the amyloid cross-ß structure. Our results indicate that hydrophobins are capable of significant conformational plasticity and the nature of the assemblies formed by these surface-active proteins is highly dependent on the interface at which self-assembly takes place.

Critical Care Management of Cerebral Edema in Brain Tumors.

Cerebral edema associated with brain tumors is extremely common and can occur in both primary and metastatic tumors. The edema surrounding brain tumors results from leakage of plasma across the vessel wall into the parenchyma secondary to disruption of the blood-brain barrier. The clinical signs of brain tumor edema depend on the location of the tumor as well as the extent of the edema, which often exceeds the mass effect induced by the tumor itself. Uncontrolled cerebral edema may result in increased intracranial pressure and acute herniation syndromes that can result in permanent neurological dysfunction and potentially fatal herniation. Treatment strategies for elevated intracranial pressure consist of general measures, medical interventions, and surgery. Alhough the definitive treatment for the edema may ultimately be surgical resection of the tumor, the impact of the critical care management cannot be underestimated and thus patients must be vigilantly monitored in the intensive care unit. In this review, we discuss the pathology, pathophysiology, and clinical features of patients presenting with cerebral edema. Imaging findings and treatment modalities used in the intensive care unit are also discussed.

Iterative projection algorithms for ab initio phasing in virus crystallography.

Iterative projection algorithms are proposed as a tool for ab initio phasing in virus crystallography. The good global convergence properties of these algorithms, coupled with the spherical shape and high structural redundancy of icosahedral viruses, allows high resolution phases to be determined with no initial phase information. This approach is demonstrated by determining the electron density of a virus crystal with 5-fold non-crystallographic symmetry, starting with only a spherical shell envelope. The electron density obtained is sufficiently accurate for model building. The results indicate that iterative projection algorithms should be routinely applicable in virus crystallography, without the need for ancillary phase information.

Predicting malignant neck lymphadenopathy using color duplex sonography based on multivariate analysis.

PURPOSE: To select the best predictors of cervical lymph node malignancy based on gray-scale and power Doppler sonography using multivariate analysis. METHODS: We evaluated sonographically a total of 97 lymph nodes in the neck that were subjected to fine-needle aspiration biopsy. The gray-scale and power Doppler sonography parameters that we analyzed using multivariate logistic regression included size, shape, echogenicity, echotexture, margins, hilum, presence of microcalcifications or necrosis, vascularization, and resistance index (RI). RESULTS: The three variables with a diagnostic accuracy exceeding 80% were an altered vascularization, heterogeneous echotexture, and abnormal hilum. Malignant nodes exhibited higher RI and larger sizes than benign nodes, and the best cutoff values to distinguish malignant from benign lymph nodes were an RI of 0.77 and a short axis ≥ 0.9 cm. Altered vascularization, a short axis ≥ 0.9 cm, and abnormal hilum were the best predictors of malignancy. CONCLUSIONS: The best sonographic predictors of lymph node malignancy are, in descending order, an altered vascularization, a short axis ≥ 0.9 cm, an abnormal hilum, and a heterogeneous echotexture. © 2016 Wiley Periodicals, Inc. J Clin Ultrasound 44:587-594, 2016.

Self-assembly of MPG1, a hydrophobin protein from the rice blast fungus that forms functional amyloid coatings, occurs by a surface-driven mechanism.

Rice blast is a devastating disease of rice caused by the fungus Magnaporthe oryzae and can result in loss of a third of the annual global rice harvest. Two hydrophobin proteins, MPG1 and MHP1, are highly expressed during rice blast infections. These hydrophobins have been suggested to facilitate fungal spore adhesion and to direct the action of the enzyme cutinase 2, resulting in penetration of the plant host. Therefore a mechanistic understanding of the self-assembly properties of these hydrophobins and their interaction with cutinase 2 is crucial for the development of novel antifungals. Here we report details of a study of the structure, assembly and interactions of these proteins. We demonstrate that, in vitro, MPG1 assembles spontaneously into amyloid structures while MHP1 forms a non-fibrillar film. The assembly of MPG1 only occurs at a hydrophobic:hydrophilic interface and can be modulated by MHP1 and other factors. We further show that MPG1 assemblies can much more effectively retain cutinase 2 activity on a surface after co-incubation and extensive washing compared with other protein coatings. The assembly and interactions of MPG1 and MHP1 at hydrophobic surfaces thereby provide the basis for a possible mechanism by which the fungus can develop appropriately at the infection interface.