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Full-Text Articles in Social and Behavioral Sciences

Integrated Analysis Of Content And Construct Validity, Byron Gajewski, Larry Price, Valorie Coffland, Diane Boyle, Marjorie Bott Apr 2015

Integrated Analysis Of Content And Construct Validity, Byron Gajewski, Larry Price, Valorie Coffland, Diane Boyle, Marjorie Bott

Diane Kay Boyle PhD, RN, FAAN

Establishing adequacy of psychometric properties of an instrument involves acquisition and evaluation of evidence based on item content and internal structure. Content validity evidence consists of subject matter experts providing quantitative ratings of the extent to which items are a representative sample of targeted domain. Evidence of internal structure includes factor analytic studies and examination of item interrelationships based on item responses from participants. Although subject matter expert ratings and participant response data are traditionally analyzed separately, each serves to inform the other in important ways. We propose integrating subject matter experts’ and participants’ data seamlessly to establish a unified …


A Review Of Baobab (Adansonia Digitata) Products: Effect Of Processing Techniques, Medicinal Properties And Uses, Donatien Kabore, Hagrétou Sawadogo-Lingani, Bréhima Diawara, Clarise Compaoré, Mamoudou H. Dicko Prof., Mogens Jacobsen Dec 2011

A Review Of Baobab (Adansonia Digitata) Products: Effect Of Processing Techniques, Medicinal Properties And Uses, Donatien Kabore, Hagrétou Sawadogo-Lingani, Bréhima Diawara, Clarise Compaoré, Mamoudou H. Dicko Prof., Mogens Jacobsen

Pr. Mamoudou H. DICKO, PhD

A general literature review including the effect of processing techniques, medicinal value and uses of baobab tree is reported in this manuscript. Baobab tree has multi-purpose uses, as it produces food and non-food products such as medicines, fuel, timber, fodder. Every part of the baobab tree is reported to be useful. The seeds, leaves, roots, flowers, fruit pulp and bark of baobab are edible. Baobab leaves are used in the preparation of soup. Seeds are used as a thickening agent in soups, but they can be fermented and used as a flavouring agent or roasted and eaten as snacks. The …


The Massachusetts Environmental Industry: Facing The Challenges Of Maturity, Betty J. Diener, David Terkla, Erick Cooke Dec 2011

The Massachusetts Environmental Industry: Facing The Challenges Of Maturity, Betty J. Diener, David Terkla, Erick Cooke

David G. Terkla

For most of the past 20 years, the environmental industry has been a very significant one, both in Massachusetts and across the country. Some have placed it alongside the electronics, computer hardware, software, biotechnology, fiber optics, and composite materials industries as part of the high-technology sector that has diversified and strengthened the state’s economy. Nationally, environmental industry employment exceeded that of several major manufacturing industries, including chemicals, paper, and aerospace. In the late 1990s, however, the momentum of the environmental movement began to wane. A decline in both employment and sales suggests that many of the most pressing environmental concerns …


2010 Massachusetts Recreational Boater Survey: Final Report Submitted To The Massachusetts Ocean Partnership, Dan Hellin, Jack Wiggin, Kristin Uiterwyk, Kim Starbuck, Nicholas Napoli, David Terkla, Chris Watson, Anthony Roman, Leona Roach, Tim Welch Dec 2011

2010 Massachusetts Recreational Boater Survey: Final Report Submitted To The Massachusetts Ocean Partnership, Dan Hellin, Jack Wiggin, Kristin Uiterwyk, Kim Starbuck, Nicholas Napoli, David Terkla, Chris Watson, Anthony Roman, Leona Roach, Tim Welch

David G. Terkla

The Massachusetts Ocean Management Plan (Plan) completed in 2009 recognized recreational boating as an activity with “significant actual and prospective conflicts among multiple waterway uses in Massachusetts” and included the economic value of recreational boating as a key socio-economic indicator that will be used to inform coastal management. At the time of Plan completion, statistically robust recreational boating data were identified as an important need for comprehensive ocean planning. To fill this data gap, the 2010 Massachusetts Recreational Boater Survey gathered information on boating activity in Massachusetts’ coastal and ocean waters directly from recreational boaters. Researchers sent 10,000 surveys to …


Model For Amorphous Aggregation Processes, Samual Stranks, Heath Ecroyd, Steve Van Sluyter, Elizabeth J. Waters, John A. Carver, Lorenz Von Smekal Dec 2011

Model For Amorphous Aggregation Processes, Samual Stranks, Heath Ecroyd, Steve Van Sluyter, Elizabeth J. Waters, John A. Carver, Lorenz Von Smekal

Heath Ecroyd

The amorphous aggregation of proteins is associated with many phenomena, ranging from the formation of protein wine haze to the development of cataract in the eye lens and the precipitation of recombinant proteins during their expression and purification. While much literature exists describing models for linear protein aggregation, such as amyloid fibril formation, there are few reports of models which address amorphous aggregation. Here, we propose a model to describe the amorphous aggregation of proteins which is also more widely applicable to other situations where a similar process occurs, such as in the formation of colloids and nanoclusters. As first …


Endogenous Redox Activity In Mouse Spermatozoa And Its Role In Regulating The Tyrosine Phosphorylation Events Associated With Sperm Capacitation, Heath W. Ecroyd, Russell C. Jones, Robert J. Aitken Dec 2011

Endogenous Redox Activity In Mouse Spermatozoa And Its Role In Regulating The Tyrosine Phosphorylation Events Associated With Sperm Capacitation, Heath W. Ecroyd, Russell C. Jones, Robert J. Aitken

Heath Ecroyd

We investigated the role of endogenous redox activity in regulating the signal transduction pathway leading to tyrosine phosphorylation in mouse spermatozoa. Endogenous redox activity was monitored using a luminol-peroxidase chemiluminescent probe. Chemiluminescence increased in spermatozoa that were actively undergoing cAMP-mediated tyrosine phosphorylation events associated with capacitation and was inhibited in a dose-dependent manner by addition of catalase or diphenylene iodonium, both of which also inhibited tyrosine phosphorylation within the cell at points downstream of cAMP. Excluding bicarbonate from the incubation medium reduced the redox activity of sperm by 80-90% and dramatically reduced tyrosine phosphorylation. This study provides the first evidence …


Role Of The Epididymis In Sperm Competition, Russell Jones, Jl Dacheux, Brett Nixon, Heath Ecroyd Dec 2011

Role Of The Epididymis In Sperm Competition, Russell Jones, Jl Dacheux, Brett Nixon, Heath Ecroyd

Heath Ecroyd

Although it is generally understood that the testes recruited kidney ducts for reproductive function during the evolution of vertebrates, little is understood of the biological significance of the adaptation. In the context of the evolution of the mammalian epididymis, this report provides evidence that a major role of the epididymis is to enhance a male's chance of achieving paternity in a competitive mating system. A unique example of sperm cooperation, in monotremes is used as evidence that the epididymis produces sperm competition proteins to form groups of 100 sperm into bundles that have a forward motility nearly thrice that of …


Carboxymethylated-K-Casein: A Convenient Tool For The Identification Of Polyphenolic Inhibitors Of Amyloid Fibril Formation, John A. Carver, Peter J. Duggan, Heath Ecroyd, Yanqin Liu, Adam G. Meyer, C E. Tranberg Dec 2011

Carboxymethylated-K-Casein: A Convenient Tool For The Identification Of Polyphenolic Inhibitors Of Amyloid Fibril Formation, John A. Carver, Peter J. Duggan, Heath Ecroyd, Yanqin Liu, Adam G. Meyer, C E. Tranberg

Heath Ecroyd

Reduced and carboxymethylated-κ-casein (RCM-κ-CN) is a milk-derived amyloidogenic protein that readily undergoes nucleation-dependent aggregation and amyloid fibril formation via a similar pathway to disease-specific amyloidogenic peptides like amyloid beta (Aβ), which is associated with Alzheimer’s disease. In this study, a series of flavonoids, many known to be inhibitors of Aβ fibril formation, were screened for their ability to inhibit RCM-κ-CN fibrilisation, and the results were compared with literature data on Aβ inhibition. Flavonoids that had a high degree of hydroxylation and molecular planarity gave good inhibition of RCM-κ-CN fibril formation. IC50 values were between 10- and 200-fold higher with RCM-κ-CN …


Tyrosine Phosphorylation Of Hsp-90 During Mammalian Sperm Capacitation, Heath Ecroyd, Russell Jones, Robert Aitken Dec 2011

Tyrosine Phosphorylation Of Hsp-90 During Mammalian Sperm Capacitation, Heath Ecroyd, Russell Jones, Robert Aitken

Heath Ecroyd

The process of sperm capacitation is correlated with activation of a signal transduction pathway leading to protein tyrosine phosphorylation. Whereas phosphotyrosine expression is an essential prerequisite for fertilization, the proteins that are phosphorylated during capacitation have not yet been identified. In the present study, we observed that a major target of this signaling pathway is the molecular chaperone protein, heat shock protein (HSP)-86, a member of the HSP-90 family of HSPs. We used cross-immunoprecipitation experiments to confirm the tyrosine phosphorylation of HSP-86, a process that is not inhibited by the ansamycin antibiotic, geldanamycin. The general significance of these findings was …


Dissociation From The Oligomeric State Is The Rate-Limiting Step In Fibril Formation By Kappa-Casein, Heath Ecroyd, Tomas Koudelka, David Thorn, Danielle Williams, Glyn Devlin, Peter Hoffmann, John Carver Dec 2011

Dissociation From The Oligomeric State Is The Rate-Limiting Step In Fibril Formation By Kappa-Casein, Heath Ecroyd, Tomas Koudelka, David Thorn, Danielle Williams, Glyn Devlin, Peter Hoffmann, John Carver

Heath Ecroyd

Amyloid fibrils are aggregated and precipitated forms of protein in which the protein exists in highly ordered, long, unbranching threadlike formations that are stable and resistant to degradation by proteases. Fibril formation is an ordered process that typically involves the unfolding of a protein to partially folded states that subsequently interact and aggregate through a nucleation-dependent mechanism. Here we report on studies investigating the molecular basis of the inherent propensity of the milk protein, kappa-casein, to form amyloid fibrils. Using reduced and carboxymethylated kappa-casein ( RCM kappa-CN), we show that fibril formation is accompanied by a characteristic increase in thioflavin …


An Epididymal Form Of Cauxin, A Carboxylesterase-Like Enzyme, Is Present And Active In Mammalian Male Reproductive Fluids, Heath W. Ecroyd, Maya Belghazi, Jl Dacheux, M Miyazaki, T Yamashita, Jl Gatti Dec 2011

An Epididymal Form Of Cauxin, A Carboxylesterase-Like Enzyme, Is Present And Active In Mammalian Male Reproductive Fluids, Heath W. Ecroyd, Maya Belghazi, Jl Dacheux, M Miyazaki, T Yamashita, Jl Gatti

Heath Ecroyd

Mass spectrometric analysis of a prion protein (PrP)-containing complex isolated from ram cauda epididymal fluid revealed a protein that showed homology to a carboxylesterase-like protein previously identified in cat urine (cauxin). Using anticauxin antibodies, immunoreactive bands were detected in corpus and cauda epididymal fluid from all mammals tested (ram, boar, mouse, and cat). In the ram, the protein was also present in seminal fluid but not found to be associated with sperm. The bands reacting with the anti-cauxin antibody coincided with those having esterase activity in a zymographic assay and its levels paralleled the esterase activity of native epididymal fluids. …


The Dissociated Form Of Kappa-Casein Is The Precursor To Its Amyloid Fibril Formation, Heath Ecroyd, David Thorn, Yanqin Liu, John Carver Dec 2011

The Dissociated Form Of Kappa-Casein Is The Precursor To Its Amyloid Fibril Formation, Heath Ecroyd, David Thorn, Yanqin Liu, John Carver

Heath Ecroyd

Bovine milk kappa-casein forms a self-associating oligomeric micelle-like species, in equilibrium with dissociated forms. In its native form, intra- and inter-molecular disulfide bonds lead to the formation of multimeric species ranging from monomers to decamers. When incubated under conditions of physiological pH and temperature, both reduced and non-reduced kappa-casein form highly structured beta-sheet amyloid fibrils. We investigated whether the precursor to kappa-casein fibril formation is a dissociated state of the protein or its oligomeric micelle-like form. We show that reduced kappa-casein is capable of forming fibrils well below its critical micelle concentration, i.e. at concentrations where only dissociated forms of …


The Effect Of Small Molecules In Modulating The Chaperone Activity Of Alpha B-Crystallin Against Ordered And Disordered Protein Aggregation, Heath Ecroyd, John Carver Dec 2011

The Effect Of Small Molecules In Modulating The Chaperone Activity Of Alpha B-Crystallin Against Ordered And Disordered Protein Aggregation, Heath Ecroyd, John Carver

Heath Ecroyd

Protein aggregation can proceed via disordered or ordered mechanisms, with the latter being associated with amyloid fibril formation, which has been linked to a number of debilitating conditions including Alzheimer's, Parkinson's and Creutzfeldt-Jakob diseases. Small heat-shock proteins (sHsps), such as alpha B-crystallin, act as chaperones to prevent protein aggregation and are thought to play a key role in the prevention of protein-misfolding diseases. In this study, we have explored the potential for small molecules such as arginine and guanidine to affect the chaperone activity of alpha B-crystallin against disordered (amorphous) and ordered (amyloid fibril) forms of protein aggregation. The effect …


The Interaction Of Unfolding Α-Lactalbumin And Malate Dehydrogenase With The Molecular Chaperone Αb-Crystallin: A Light And X-Ray Scattering Investigation, J W. Regini, Heath Ecroyd, Sarah Meehan, Kristen Bremmell, Matthew J. Clarke, Donna Lammie, Tim Wess, John A. Carver Dec 2011

The Interaction Of Unfolding Α-Lactalbumin And Malate Dehydrogenase With The Molecular Chaperone Αb-Crystallin: A Light And X-Ray Scattering Investigation, J W. Regini, Heath Ecroyd, Sarah Meehan, Kristen Bremmell, Matthew J. Clarke, Donna Lammie, Tim Wess, John A. Carver

Heath Ecroyd

Purpose: The molecular chaperone αB-crystallin is found in high concentrations in the lens and is present in all major body tissues. Its structure and the mechanism by which it protects its target protein from aggregating and precipitating are not known. Methods: Dynamic light scattering and X-ray solution scattering techniques were used to investigate structural features of the αB-crystallin oligomer when complexed with target proteins under mild stress conditions, i.e., reduction of α- lactalbumin at 37 °C and malate dehydrogenase when heated at 42 °C. In this investigation, the size, shape and particle distribution of the complexes were determined in real-time …


Analysis Of The Mechanism By Which Calcium Negatively Regulates The Tyrosine Phosphorylation Cascade Associated With Sperm Capacitation, Mark Baker, Louise Hethrington, Heath Ecroyd, Shaun Roman, Robert Aitken Dec 2011

Analysis Of The Mechanism By Which Calcium Negatively Regulates The Tyrosine Phosphorylation Cascade Associated With Sperm Capacitation, Mark Baker, Louise Hethrington, Heath Ecroyd, Shaun Roman, Robert Aitken

Heath Ecroyd

The capacitation of mammalian spermatozoa involves the activation of a cAMP-mediated signal transduction pathway that drives tyrosine phosphorylation via mechanisms that are unique to this cell type. Controversy surrounds the impact of extracellular calcium on this process, with positive and negative effects being recorded in independent publications. We clearly demonstrate that the presence of calcium in the external medium decreases tyrosine phosphorylation in both human and mouse spermatozoa. Under these conditions, a rise in intracellular pH was recorded, however, this event was not responsible for the observed changes in phosphotyrosine expression. Rather, the impact of calcium on tyrosine phosphorylation in …


(-)-Epigallocatechin-3-Gallate (Egcg) Maintains K-Casein In Its Pre-Fibrillar State Without Redirecting Its Aggregation Pathway, Sean A. Hudson, Heath Ecroyd, Francis C. Dehle, Ian F. Musgrave, John Carver Dec 2011

(-)-Epigallocatechin-3-Gallate (Egcg) Maintains K-Casein In Its Pre-Fibrillar State Without Redirecting Its Aggregation Pathway, Sean A. Hudson, Heath Ecroyd, Francis C. Dehle, Ian F. Musgrave, John Carver

Heath Ecroyd

The polyphenol (-)-epigallocatechin-3-gallate (EGCG) has recently attracted much research interest in the field of protein-misfolding diseases because of its potent anti-amyloid activity against amyloid-beta, alpha-synuclein and huntingtin, the amyloid-fibril-forming proteins involved in Alzheimer's, Parkinson's and Huntington's diseases, respectively. EGCG redirects the aggregation of these polypeptides to a disordered off-folding pathway that results in the formation of non-toxic amorphous aggregates. whether this anti-fibril activity is specific to these disease-related target proteins or ismore generic remains to be established. In addition, the mechanism by which EGCG exerts its effects, as with all anti-amyloidogenic polyphenols, remains unclear. To address these aspects, we have …


The Two Faced Nature Of Milk Casein Proteins: Amyloid Fibril Formation And Chaperone-Like Activity, David Thorn, Heath Ecroyd, John Carver Dec 2011

The Two Faced Nature Of Milk Casein Proteins: Amyloid Fibril Formation And Chaperone-Like Activity, David Thorn, Heath Ecroyd, John Carver

Heath Ecroyd

Molecular chaperones are a diverse group of proteins that stabilise partially folded target proteins to prevent their misfolding, aggregation and potential precipitation under conditions of cellular stress, e.g. elevated temperature. Protein aggregation, particularly the formation of highly ordered protein aggregates termed amyloid fibrils, is of considerable research interest because of its intimate association with a wide range of debilitating diseases, including Alzheimer's, Parkinson's and Huntington's diseases and type II diabetes. In this review, we discuss the ability of the milk casein proteins to act in a chaperone-like manner. This property is of biological importance since at least two of the …


New Proteins Identified In Epididymal Fluid From The Platypus (Ornithorhynchus Anatinus), Jean-Louis Dacheux, Francoise Dacheux, Valerie Labas, Heath Ecroyd, Brett Nixon, Russell C. Jones Dec 2011

New Proteins Identified In Epididymal Fluid From The Platypus (Ornithorhynchus Anatinus), Jean-Louis Dacheux, Francoise Dacheux, Valerie Labas, Heath Ecroyd, Brett Nixon, Russell C. Jones

Heath Ecroyd

The platypus epididymal proteome is being studied because epididymal proteins are essential for male fertility in mammals and it is considered that knowledge of the epididymal proteome in an early mammal would be informative in assessing the convergence and divergence of proteins that are important in the function of the mammalian epididymis. Few of the epididymal proteins that have been identified in eutherian mammals were found in platypus caudal epididymal fluid, and the major epididymal proteins in the platypus (PXN-FBPL, SPARC and E-OR20) have never been identified in the epididymis of any other mammal.


Mimicking Phosphorylation Of Alphab-Crystallin Affects Its Chaperone Activity, Heath W. Ecroyd, Sarah Meehan, J Horwitz, Andrew Aquilina, J L Benesch, C V Robinson, Cait Macphee, John Carver Dec 2011

Mimicking Phosphorylation Of Alphab-Crystallin Affects Its Chaperone Activity, Heath W. Ecroyd, Sarah Meehan, J Horwitz, Andrew Aquilina, J L Benesch, C V Robinson, Cait Macphee, John Carver

Heath Ecroyd

No abstract provided.


The Thioflavin T Fluorescence Assay For Amyloid Fibril Detection Can Be Biased By The Presence Of Exogenous Compounds, Sean A. Hudson, Heath Ecroyd, Tak W. Kee, John A. Carver Dec 2011

The Thioflavin T Fluorescence Assay For Amyloid Fibril Detection Can Be Biased By The Presence Of Exogenous Compounds, Sean A. Hudson, Heath Ecroyd, Tak W. Kee, John A. Carver

Heath Ecroyd

Thioflavin T (ThT) dye fluorescence is used regularly to quantify the formation and inhibition of amyloid fibrils in the presence of anti-amyloidogenic compounds such as polyphenols. However, in this study, it was shown, using three polyphenolics (curcumin, quercetin and resveratrol), that ThT fluorescence should be used with caution in the presence of such exogenous compounds. The strong absorptive and fluorescent properties of quercetin and curcumin were found to significantly bias the ThT fluorescence readings in both in situ real-time ThT assays and single time-point dilution ThT-type assays. The presence of curcumin at concentrations as low as 0.01 and 1 uM …


Unraveling The Mysteries Of Protein Folding And Misfolding, Heath Ecroyd, John A. Carver Dec 2011

Unraveling The Mysteries Of Protein Folding And Misfolding, Heath Ecroyd, John A. Carver

Heath Ecroyd

This mini-review focuses on the processes and consequences of protein folding and misfolding. The latter process often leads to protein aggregation and precipitation with the aggregates adopting either highly ordered (amyloid fibril) or disordered (amorphous) forms. In particular, the amyloid fibril is discussed because this form has gained considerable notoriety due to its close links to a variety of debilitating diseases including Alzheimer's, Parkinson's, Huntington's, and Creutzfeldt-Jakob diseases, and type-II diabetes. In each of these diseases a different protein forms fibrils, yet the fibrils formed have a very similar structure. The mechanism by which fibrils form, fibril structure, and the …


Crystallin Proteins And Amyloid Fibrils, Heath Ecroyd, John A. Carver Dec 2011

Crystallin Proteins And Amyloid Fibrils, Heath Ecroyd, John A. Carver

Heath Ecroyd

Improper protein folding (misfolding) can lead to the formation of disordered (amorphous) or ordered (amyloid fibril) aggregates. The major lens protein, alpha-crystallin, is a member of the small heat-shock protein (sHsp) family of intracellular molecular chaperone proteins that prevent protein aggregation. Whilst the chaperone activity of sHsps against amorphously aggregating proteins has been well studied, its action against fibril-forming proteins has received less attention despite the presence of sHsps in deposits found in fibril-associated diseases (e.g. Alzheimer's and Parkinson's). In this review, the literature on the interaction of alpha B-crystallin and other sHsps with fibril-forming proteins is summarized. In particular, …


Amyloid Fibril Formation By Bovine Milk Alpha(S2)-Casein Occurs Under Physiological Conditions Yet Is Prevented By Its Natural Counterpart, Alpha(S1)-Casein, David Thorn, Heath Ecroyd, M Sunde, Stephen Poon, John Carver Dec 2011

Amyloid Fibril Formation By Bovine Milk Alpha(S2)-Casein Occurs Under Physiological Conditions Yet Is Prevented By Its Natural Counterpart, Alpha(S1)-Casein, David Thorn, Heath Ecroyd, M Sunde, Stephen Poon, John Carver

Heath Ecroyd

The calcified proteinaceous deposits, or corpora amylacea, of bovine mammary tissue often comprise a network of amyloid fibrils, the origins of which have not been fully elucidated. Here, we demonstrate by transmission electron microscopy, dye binding assays, and X-ray fiber diffraction that bovine milk alpha(s2)-casein, a protein synthesized and secreted by mammary epithelial cells, readily forms fibrils in vitro. As a component of whole alpha(s)-casein, alpha(s2)-casein was separated from alpha(s1)-casein under nonreducing conditions via cation-exchange chromatography. Upon incubation at neutral pH and 37 degrees C, the spherical particles typical of alpha(s2)-casein rapidly converted to twisted, ribbon-like fibrils similar to 12 …


Alphab-Crystallin Inhibits The Cell Toxicity Associated With Amyloid Fibril Formation By Kappa-Casein And The Amyloid-Beta Peptide, Francis C. Dehle, Heath Ecroyd, Ian F. Musgrave, John A. Carver Dec 2011

Alphab-Crystallin Inhibits The Cell Toxicity Associated With Amyloid Fibril Formation By Kappa-Casein And The Amyloid-Beta Peptide, Francis C. Dehle, Heath Ecroyd, Ian F. Musgrave, John A. Carver

Heath Ecroyd

Amyloid fibril formation is associated with diseases such as Alzheimer’s, Parkinson’s, and prion diseases. Inhibition of amyloid fibril formation by molecular chaperone proteins, such as the small heat-shock protein αB-crystallin, may play a protective role in preventing the toxicity associated with this form of protein misfolding. Reduced and carboxymethylated κ-casein (RCMκ-CN), a protein derived from milk, readily and reproducibly forms fibrils at physiological temperature and pH. We investigated the toxicity of fibril formation by RCMκ-CN using neuronal model PC12 cells and determined whether the inhibition of fibril formation altered its cell toxicity. To resolve ambiguities in the literature, we also …


The Epididymal Soluble Prion Protein Forms A High-Molecular-Mass Complex In Association With Hydrophobic Proteins, Heath W. Ecroyd, Maya Belghazi, Jean-Louis Dacheux, Jean-Luc Gatti Dec 2011

The Epididymal Soluble Prion Protein Forms A High-Molecular-Mass Complex In Association With Hydrophobic Proteins, Heath W. Ecroyd, Maya Belghazi, Jean-Louis Dacheux, Jean-Luc Gatti

Heath Ecroyd

We have shown previously that a 'soluble' form of PrP (prion protein), not associated with membranous vesicles, exists in the male reproductive fluid [Ecroyd, Sarradin, Dacheux and Gatti (2004) Biol. Reprod. 71, 993-1001]. Attempts to purify this 'soluble' PrP indicated that it behaves like a high-molecular-mass complex of more than 350 kDa and always co-purified with the same set of proteins. The main associated proteins were sequenced by MS and were found to match to clusterin (apolipoprotein J), BPI (bacterial permeability-increasing protein), carboxylesterase-like urinary excreted protein (cauxin), beta-mannosidase and beta-galactosidase. Immunoblotting and enzymatic assay confirmed the presence of clusterin and …


Compartmentalization Of Prion Isoforms Within The Reproductive Tract Of The Ram, Heath W. Ecroyd, Pierre Sarradin, Jean-Louis Dacheux, Jean-Luc Gatti Dec 2011

Compartmentalization Of Prion Isoforms Within The Reproductive Tract Of The Ram, Heath W. Ecroyd, Pierre Sarradin, Jean-Louis Dacheux, Jean-Luc Gatti

Heath Ecroyd

Cellular prion protein (Prp(C)) is a glycoprotein usually associated with membranes via its glycosylphosphatidylinositol (GPI) anchor. The trans-conformational form of this protein (Prp(SC)) is the suggested agent responsible for transmissible neurodegenerative spongiform encephalopathies. This protein has been shown on sperm and in the reproductive fluids of males. Antibodies directed against the C-terminal sequence near the GPI-anchor site, an N-terminal sequence, and against the whole protein showed that the Prp isoforms were compartmentalized within the reproductive tract of the ram. Immunoblotting with the three antibodies showed that the complete protein and both N- and C-terminally truncated and glycosylated isoforms are present …


Post-Testicular Sperm Environment And Fertility, Jean-Luc Gatti, Sandrine Castella, Francoise Dacheux, Heath Ecroyd, S Metayer, Veronique Thimon, Jean-Louis Dacheux Dec 2011

Post-Testicular Sperm Environment And Fertility, Jean-Luc Gatti, Sandrine Castella, Francoise Dacheux, Heath Ecroyd, S Metayer, Veronique Thimon, Jean-Louis Dacheux

Heath Ecroyd

When mammalian spermatozoa exit the testis, they show a highly specialized morphology; however, they are not yet able to carry out their task: to fertilize an oocyte. This property, that includes the acquisition of motility and the ability to recognize and to fuse with the oocyte investments, is gained only after a transit through the epididymis during which the spermatozoa from the testis travel to the vas deferens. The exact molecular mechanisms that turn these cells into fertile gametes still remain mysterious, but surface-modifying events occurring in response to the external media are key steps in this process. Our laboratory …


Protective Interactions Of Dairy Peptides With Fibril Structures And Relevance To Alzheimer's Disease, Louise Bennett, Williams Roderick, Heath Ecroyd, Yanqin Liu, Sunanda Sudharmarajan, John Carver Dec 2011

Protective Interactions Of Dairy Peptides With Fibril Structures And Relevance To Alzheimer's Disease, Louise Bennett, Williams Roderick, Heath Ecroyd, Yanqin Liu, Sunanda Sudharmarajan, John Carver

Heath Ecroyd

Selected dairy caseins have been shown to have capacity for chaperone-like regulation of folding pathways of other caseins, specifically in preventing development of fibrillar aggregates of beta sheet structure. An assay based on fibril formation by reduced and carboxymethylated-kappa casein (RCM-kCn) was thus used to screen for anti-fibril activity among a selection of dairy protein hydrolysates, in order to discover peptides with possible anti-fibril, chaperone activity. From the selection of eight dairy hydrolysates based on different dairy protein fractions, two of the hydrolysates of whey protein exhibited superior anti-fibril bioactivity against RCM-kCn and amyloid beta (Aβ), the peptide associated with …


Small Heat-Shock Proteins Interact With A Flanking Domain To Suppress Polyglutamine Aggregation, Amy L. Roberston, Stephen J. Headey, Helen M. Saunders, Heath Ecroyd, Martin J. Scanlon, John A. Carver, Stephen P. Bottomley Dec 2011

Small Heat-Shock Proteins Interact With A Flanking Domain To Suppress Polyglutamine Aggregation, Amy L. Roberston, Stephen J. Headey, Helen M. Saunders, Heath Ecroyd, Martin J. Scanlon, John A. Carver, Stephen P. Bottomley

Heath Ecroyd

Small heat-shock proteins (sHsps) are molecular chaperones that play an important protective role against cellular protein misfolding by interacting with partially unfolded proteins on their off-folding pathway, preventing their aggregation. Polyglutamine (polyQ) repeat expansion leads to the formation of fibrillar protein aggregates and neuronal cell death in nine diseases, including Huntington disease and the spinocerebellar ataxias (SCAs). There is evidence that sHsps have a role in suppression of polyQ-induced neurodegeneration; for example, the sHsp alphaB-crystallin (αB-c) has been identified as a suppressor of SCA3 toxicity in a Drosophila model. However, the molecular mechanism for this suppression is unknown. In this …


Testicular Descent, Sperm Maturation And Capacitation. Lessons From Our Most Distant Relatives, The Monotremes, Russell Jones, Heath Ecroyd, Jean-Louis Dacheux, Brett Nixon Dec 2011

Testicular Descent, Sperm Maturation And Capacitation. Lessons From Our Most Distant Relatives, The Monotremes, Russell Jones, Heath Ecroyd, Jean-Louis Dacheux, Brett Nixon

Heath Ecroyd

The present review examines whether monotremes may help to resolve three questions relating to sperm production in mammals: why the testes descend into a scrotum in most mammals, why spermatozoa are infertile when they leave the testes and require a period of maturation in the specific milieu provided by the epididymides, and why ejaculated spermatozoa cannot immediately fertilise an ovum until they undergo capacitation within the female reproductive tract. Comparisons of monotremes with other mammals indicate that there is a need for considerable work on monotremes. It is hypothesised that testicular descent should be related to epididymal differentiation. Spermatozoa and …