Copper trafficking in eukaryotic systems: current knowledge from experimental and computational efforts

Copper plays a vital role in fundamental cellular functions, and its concentration in the cell must be tightly regulated, as dysfunction of copper homeostasis is linked to severe neurological diseases and cancer. This review provides a compendium of current knowledge regarding the mechanism of copper transfer from the blood system to the Golgi apparatus; this mechanism involves the copper transporter hCtr1, the metallochaperone Atox1, and the ATPases ATP7A/B. We discuss key insights regarding the structural and functional properties of the hCtr1-Atox1-ATP7B cycle, obtained from diverse studies relying on distinct yet complementary biophysical, biochemical, and computational methods. We further address the mechanistic aspects of the cycle that continue to remain elusive. These knowledge gaps must be filled in order to be able to harness our understanding of copper transfer to develop therapeutic approaches with the capacity to modulate copper metabolism

Unraveling the impact of cysteine-to-serine mutations on the structural and functional properties of Cu(I)-binding proteins

Appropriate maintenance of Cu(I) homeostasis is an essential requirement for proper cell function because its misregulation induces the onset of major human diseases and mortality. For this reason, several research efforts have been devoted to dissecting the inner working mechanism of Cu(I)-binding proteins and transporters. A commonly adopted strategy relies on mutations of cysteine residues, for which Cu(I) has an exquisite complementarity, to serines. Nevertheless, in spite of the similarity between these two amino acids, the structural and functional impact of serine mutations on Cu(I)-binding biomolecules remains unclear. Here, we applied various biochemical and biophysical methods, together with all-atom simulations, to investigate the effect of these mutations on the stability, structure, and aggregation propensity of Cu(I)-binding proteins, as well as their interaction with specific partner proteins. Among Cu(I)-binding biomolecules, we focused on the eukaryotic Atox1-ATP7B system, and the prokaryotic CueR metalloregulator. Our results reveal that proteins containing cysteine-to-serine mutations can still bind Cu(I) ions; however, this alters their stability and aggregation propensity. These results contribute to deciphering the critical biological principles underlying the regulatory mechanism of the in-cell Cu(I) concentration, and provide a basis for interpreting future studies that will take advantage of cysteine-to-serine mutations in Cu(I)-binding systems

Triangular Ring Resonator: Direct measurement of the parity-odd parameters of the photon sector of SME

We introduce the the Triangular Ring (TR) resonator. We show that the difference between the clockwise and anti-clockwise resonant frequencies of a vacuum TR resonator is sensitive to the birefringence parity-odd parameters of the photon's sector of the minimal Standard Model Extension (mSME): the Standard Model plus all the perturbative parameters encoding the break the Lorentz symmetry. We report that utilizing the current technology allows for direct measurement of these parameters with a sensitivity of the parity even ones and improves the best current resonator bounds by couple of orders of magnitudes. We note that designing an optical table that rotates perpendicular to the gravitational equipotential surface (geoid) allows for direct measurement of the constancy of the light speed at the vicinity of the earth in all directions in particular perpendicular to the geoid. If this table could achieve the precision of the ordinary tables, then it would improve the GPS bounds on the constancy of the light speed perpendicular to geoid by about eight orders of magnitude.Comment: ref. added, minor corrections, matches the published versio

Phenomenological covariant approach to gravity

We covariantly modify the Einstein-Hilbert action such that the modified action perturbatively resolves the flat rotational velocity curve of the spiral galaxies and gives rise to the Tully-Fisher relation, and dynamically generates the cosmological constant. This modification requires introducing just a single new universal parameter.Comment: v6: a mistake in deriving the equation of the cosmological constant corrected, refs adde

Constraints on f(RijklRijkl)f(R_{ijkl}R^{ijkl}) gravity: An evidence against the covariant resolution of the Pioneer anomaly

We consider corrections in the form of $\Delta L(R_{ijkl}R^{ijkl})$ to the Einstein-Hilbert Lagrangian. Then we compute the corrections to the Schwarszchild geometry due to the inclusion of this general term to the Lagrangian. We show that $\Delta L_3=\alpha_{{1/3}}(R_{ijkl}R^{ijkl})^{{1/3}}$ gives rise to a constant anomalous acceleration for objects orbiting the Sun onward the Sun. This leads to the conclusion that $\alpha_{{1/3}}=(13.91\pm 2.11) \times 10^{-26}(\frac{1}{\text{meters}})^{{2/3}}$ would have covariantly resolved the Pioneer anomaly if this value of $\alpha_{{1/3}}$ had not contradicted other observations. We notice that the experimental bounds on $\Delta L_3$ grows stronger in case we examine the deformation of the space-time geometry around objects lighter than the Sun. We therefore use the high precision measurements around the Earth (LAGEOS and LLR) and obtain a very strong constraint on the corrections in the form of $\Delta L(R_{ijkl}R^{ijkl})$ and in particular $\Delta L=\alpha_n(R_{ijkl}R^{ijkl})^n$. This bound requires $\alpha_{{1/3}}\leq6.12\times 10^{-29}(\frac{1}{\text{meters}})^{{2/3}}$. Therefore it refutes the covariant resolution of the Pioneer anomaly.Comment: ...v5: references added, new discussions adde

Amidolysis of Oxirane: Effect of Protein Type, Oils, and ZnCl 2

Amidolysis of oxirane group of epoxidized sesame, sunflower, and cottonseed oils was achieved by reaction with primary amide of millet and gluten proteins. Gluten is a coproduct of wheat starch industry and available commercially. Millet is a major part of the staple food of the semiarid region of the tropics. Gluten is a mixture of glutenins and gliadins rich in glutamine residues; however, millet is rich in glutamine and leucine. We have taken advantage of the available primary amide of glutamine for cross-linking with the oxirane of sunflower, sesame, and cottonseed oils under controlled conditions to give a resin of amidohydroxy of gluten and millet proteins. Cross-linking gave a resin with a wide range of textural properties. The texture of the resin was dependent on the source of the oxirane, the amide group, and the amount of the catalyst (ZnCl2). The thermal properties, textural, solubility, and rheological properties were determined as well as the reaction time. The data showed direct relationships between the ZnCl2, nature of oil, and protein type and the properties of the final resin. Consistently, the results pointed to similarity among the outcome of the reactions between sesame and sunflower oils. Depending on the amount of ZnCl2, the texture of the resin can range from viscose to rubbery. The reaction time was influenced by oxirane source, protein type, and catalyst and ranged from 30 min to 4 hr

Cu(I) Controls Conformational States in Human Atox1 Metallochaperone: An EPR and Multiscale Simulation Study

Atox1 is a human copper metallochaperone that is responsible for transferring copper ions from the main human copper transporter, hCtr1, to ATP7A/B in the Golgi apparatus. Atox1 interacts with the Ctr1 C-terminal domain as a dimer, although it transfers the copper ions to ATP7A/B in a monomeric form. The copper binding site in the Atox1 dimer involves Cys12 and Cys15, while Lys60 was also suggested to play a role in the copper binding. We recently showed that Atox1 can adopt various conformational states, depending on the interacting protein. In the current study, we apply EPR experiments together with hybrid quantum mechanics-molecular mechanics molecular dynamics simulations using a recently developed semiempirical density functional theory approach, to better understand the effect of Atox1's conformational states on copper coordination. We propose that the flexibility of Atox1 occurs owing to protonation of one or more of the cysteine residues, and that Cys15 is an important residue for Atox1 dimerization, while Cys12 is a critical residue for Cu(I) binding. We also show that Lys60 electrostatically stabilizes the Cu(I)-Atox1 dimer

Treatment of psychological morbidity secondary to benign prostatic hyperplasia: a comparative study

Background: Authors tend to compare the medical treatment of benign prostatic hyperplasia with the surgical option regarding lower urinary tract symptoms (LUTS) and related psychological morbidity.Methods: A retrospective study of (1614) patients who were managed by either transurethral resection of prostate (TURP) or medical treatment for lower urinary tract symptoms secondary to benign prostatic hyperplasia (BPH) over a period of 5 years between (Sep. 2013 and Sep. 2018) carried out in Prince Hussein Urology Center at Jordanian Royal Medical Services. Patients were classified into two groups, group1 (807 patients) who get a medical option and group 2 (807 patients who underwent TURPs. A comparison between both groups according to the effect of minimizing the psychological morbidities was done over a period of 1-year follow-up after reviewing the patient’s medical records.Results: Ages of the patients for group1 and 2 were (47-68 years), (49-73 years), respectively. There were significant differences at the level of depression, anxiety and psychiatric morbidity pre-treatment between both groups p-value 0.05, but significant differences in the level of improvement after treatment between both groups and in group 2 were found, p-value <0.05.Conclusions: The severity of LUTS and psychological morbidity have a positive relationship and were higher in the pretreatment surgical group, but the effect of TURP was superior to the medical group in the management of this morbidity and its causative (LUTS)

Assessment the knowledge of mothers ‘toward home accident among children less than five years in Polytechnic University of Sulaimani

Children accidents are an essential source of death and wounds among children during the first of life. Annually, millions of children are constantly becoming handicap or blemished because of mishaps.Objective: To assess knowledge of children mothers toward home accidents in polytechnic University of Sulaimani. Methodology: A simple random sample of (110) respondents were selected.. The data collected and composed during the utilization of a created questionnaire, with two parts; the first part assessed the knowledge of children mothers with (20) yes or no questions and the second part includes the mothers' demographic attributes. A pilot study was carried out for the period of December 18th to 28th 2015 in order to conclude the questionnaire accuracy during the use of (Test – Retest). A panel of (8) experts was include in the determination of the questionnaire content validity Data were analyzed through the application of descriptive statistical data analysis approach (frequency and percentage), and inferential data analysis approach (Mean of Scores).).The results of the study revealed, the mean age of the mothers was (1.86) years, (58.2%) of them had completed secondary school, (88.2%) of mothers have more than five children, (89.1%) of mother’s were in moderate socio-economic status, (47.3%) of mothers have personal reading information,(58.2) of the child were male,(13.6%) of mothers have knowledge about safe housing condition held in Maternal and Child Health Centers.The conclusion of this study revealed that, inadequate children mothers’ knowledge regarding home accidents among children less than six years held in Maternal and Child Health Centers during the antenatal period. So the recommendation are suggested, reinforces the need for effective education program via mass media, for mothers

Development of multiplex PCR and multi-color fluorescent in situ hybridization (m-FISH) coupled protocol for detection and imaging of multi-pathogens involved in inflammatory bowel disease

Abstract Background Several pathogens have been debated to play a role in inflammatory bowel disease (IBD) including Crohn’s disease (CD). None of these pathogens have been investigated together in same clinical samples. We developed a multiplex PCR and multi-color fluorescent in situ hybridization (m-FISH) protocols for simultaneous detection of CD-associated pathogens including Mycobacterium avium subspecies paratuberculosis (MAP), Klebsiella pneumoniae, and adherent-invasive Escherichia coli strain LF82. Methods The multiplex PCR is based on 1-h DNAzol® extraction protocol modified for rapid extraction of bacterial DNA from culture, blood, and intestinal biopsies. Oligonucleotide primers sequences unique to these pathogens were evaluated individually and in combinations using bioinformatics and experimental approaches. m-FISH was based on fluorescent-tagged oligonucleotides and confocal scanning laser microscopy (CSLM). Results Following several attempts, the concentration of the oligonucleotide primers and DNA templates and the PCR annealing temperatures were optimized. Multiplex PCR analyses revealed excellent amplification signal in trials where a single primer set and combinations of two and three primers sets were tested against a mixture of DNA from three different bacteria or a mixture of three bacterial cultures mixed in one tube before DNA extraction. Slides with individual and mixtures of bacterial cultures and intestinal tissue sections from IBD patients were tested by m-FISH and the CSLM images verified multiplex PCR results detected on 3% agarose gel. Conclusion We developed a 4-h multiplex PCR protocol, which was validated by m-FISH images, capable of detecting up to four genes from major pathogens associated with CD. The new protocol should serve as an excellent tool to support efforts to study multi-pathogens involved in CD and other autoimmune disease