bluuebunny/crossref_metadata_2025_split huggingface dataset:
The role of protein oxidation in the onset of the transthyretin amyloidoses
Authors: Zhao Lei , Reixach Natalia , Buxbaum Joel 
Published: June 2021

The transthyretin (TTR) amyloidoses are a subset of protein misfolding diseases characterized by the age‐dependent deposition of amyloid fibrils, derived from either wild type or mutant forms of the plasma protein TTR. Although it is clear that age is a major risk factor for the development of the TTR and other systemic amyloidoses, there are few studies addressing the molecular mechanisms responsible for the apparent effect of aging.There is a general increase in the concentrations of oxidation products of proteins, lipids and DNA associated with aging. We hypothesize that an increase in TTR oxidation with age predisposes to TTR aggregation in vivo. We have produced oxidized forms of wild‐type TTR similar to those reported to be present in human plasma. Chaotrope‐mediated denaturation/renaturation experiments showed that modified forms of TTR, including those with oxidized methionines and cysteines and those with carbonylated amino acids displayed reduced in vitro stability relative to native non‐oxidized TTR. The modified TTRs also had differential propensity for amyloid fibril formation with respect to native TTR. Sera from mice transgenic for a wild‐type human TTR gene show increased degrees of TTR carbonylation with aging. The greater proportion of circulating oxidized TTR seen in the older mice with its lower stability, may play a role in the development and onset of the senile wild type TTR associated amyloidosis. (This work is supported by 1R01AG032285 NIH/NIA)
Structural insights into the mechanism of acetolactate decarboxylase
Authors: Marlow Victoria , Rea Dean , Wills Martin , Fülöp Vilmos 
Published: June 2021

Acetolactate decarboxylase (ALDC) is a bacterial enzyme of the butanediol fermentation pathway that decarboxylates S‐acetolactate into R‐acetoin. The remarkable feature of this enzyme is that it also catalyses the decarboxylation of the opposite enantiomer R‐acetolactate to give the same product R‐acetoin. The enzyme is widely used commercially by the brewing industry. It shortens the maturation step by preventing the formation of diacetyl, which gives an off taste to beer. The structure of Bacillus brevis ALDC has been solved using X‐ray crystallography, in the presence of a series of chiral substrate analogues. In the resting state the active site zinc ion is coordinated to three conserved histidines and a glutamate. Upon binding of the substrate analogues the C‐terminal loop moves away losing the glutamate coordination bond. The structural information combined with experimental results from active site mutants allows us to present a mechanism for the stereospecific decarboxylation of acetolactate. The structural and biochemical data we have obtained will be used to develop further modifications and applications of the enzyme in future work.The authors thank EPSRC for funding through the Molecular Organisation and Assembly in Cells (MOAC) Doctoral Training Centre.
FK962 induces neurite elongation in cultured monkey trigeminal ganglion cells
Authors: Nakajima Emi , Walkup Ryan D , Shearer Thomas R , Azuma Mitsuyoshi 
Published: June 2021

LASIK refractive surgery is widely accepted for correcting myopia. Creation of the corneal flap in LASIK surgery can lead to dry eye due to amputation of corneal sensory nerves. Previous studies in the hippocampus showed that a substituted fluorobenzamide drug, FK962, caused release of neurotrophic peptides that stimulate and control nerve activity. Our studies in rabbits also showed that FK962 accelerated extension of neuronal processes and recovery of corneal sensitivity. The purposes of the present study were to 1) determine if FK962 induces spouting and elongation of neurites in cultured monkey trigeminal ganglion (TG) cells, and 2) investigate involvement of the neurotrophic peptide GDNF in FK‐induced neurite elongation. Dissociated, cultured TG cells, containing neuronal cells and Schwann cells, were cultured for 48 hours with or without FK962. Neuronal length and elongation were evaluated by staining with antibody for neurofilament specific to neurons. Culture with GDNF, or with antibody against GDNF, was used to determine the role of GDNF in neurite elongation. Our results showed that FK962, or GDNF alone, significantly induced neurite elongation. GDNF antibody significantly inhibited neurite elongation induced by FK962 or GDNF. Our data suggested that GDNF was a mediator of FK962‐induced neurite elongation. FK962 might be a candidate drug for treatment of complications after LASIK.Dr. Shearer receives consulting fees from, and Drs. Azuma and Nakajima are employees of, Senju Pharmaceutical Co. Ltd.
Optimizing peptide yields from human hair
Authors: Steenstra Joshua Adam , Clifford Patricia , Parker Glendon 
Published: June 2021

Hair is physically robust and persists in the environment for long periods. It is a common component of crime scenes, yet it is not often analyzed because the DNA information is absent or difficult to obtain. Hair, a complex biological connection between the crime scene and the suspect, is now only rarely used in crime scene investigation. This project seeks to examine the use of protein in developing a measure of identity, we currently have a power of discrimination of 1 in 15,500. The degree of identity between an individual and a hair sample is a function of proteomic information obtained.The purpose and question of our research is to determine if there is a more efficient way to optimize protein yields from hair. To maximize the surface area to volume ratio, a hair is ground into powder. The sample is then digested and a fraction of the sample is then loaded onto a HPLC and a profile is measured, both total absorbance and peak complexity. This corresponds to how many different polypeptides and how much polypeptide is present in the sample. A range of conditions will be tested to optimize the protocol. We have already determined that mass spectrophotometry compatible detergents increase peptide yield. We anticipate developing a robust protocol that optimizes peptide yields from hair samples. This will maximize the amount of information from each sample to achieve the greatest attainable degree of identity that is consistently reliable. This methodology will prove useful in optimizing the degree of information extracted from forensically relevant material.
Targeting Angeogenesis for Bone Repair
Authors: Clemens Thomas L. 
Published: June 2021

Impaired bone healing following trauma, surgery, and/or infections continues to be challenging clinical problems. A major goal of our work is to identify the cellular and molecular mechanisms that couple angiogenesis and osteogenesis during bone regeneration, and to use this information to develop strategies to enhance bone repair. Neoangiogenesis is critical for bone regeneration and is dependent of hypoxic stimuli, which upregulate pro‐angiogenic factors including VEGF and BMP‐2. Using both genetic and pharmacologic strategies we have manipulated the VEGF and BMP‐2 pathways in mouse osteoblasts to drive angiogenesis in an attempt to enhance osteogenesis. Mice with constitutive VEGF or BMP expression in osteoblasts had markedly increased vascularity and bone regeneration, which were inhibited by concomitant administration of VEGF receptor antibodies. Administration of synthetic inhibitors of HIF prolylhydroxylation stabilized VEGF production and increased angiogenesis and bone regeneration. Additional strategies developed to increase angiogenesis involve delivering mesenchymal stromal cells transduced with a recombinant adeno‐associated viruses encoding VEGF or BMP‐2. Finally, high throughput screening approaches identified several small molecules capable activating Smad‐1 signaling downstream of the BMP receptor. These stable BMP‐activating molecules manufactured into nanoparticles can be readily delivered to sites of bone injury and are proving useful in animal models for accelerating bone repair.Supported by NIAMS R01AR49410
Molecular Fingerprinting of Hsp27 Anti‐Apoptotic Activity
Authors: Tossey Justin , Voss Oliver , Doseff Andrea 
Published: June 2021

Apoptosis is an evolutionarily conserved mechanism crucial to the homeostasis of cell numbers, tissue development, and immune response. Several checkpoints regulate the activation of apoptosis, including heat shock protein 27 (Hsp27), an anti‐apoptotic regulator that blocks several steps in the apoptotic cascade. However, the structural basis for this protective effect remains unknown.Increased cardiomyocyte (CM) apoptosis contributes to cardiovascular disease (CVD) in doxorubicin (dox)‐treated cancer patients. CM apoptosis requires the activation of caspase‐3, a key evolutionarily conserved protease. We showed that Hsp27 associates with and inhibits caspase‐3, but the exact mechanisms remain unknown. The goal of this study was to determine the structural‐functional relationships responsible for the anti‐apoptotic activity of Hsp27. Serial deletion clones of Hsp27 were used to fingerprint the Hsp27 domains responsible for the association with caspase‐3 and inhibition of apoptosis. We identified different molecular signatures responsible for caspase‐3 association and the regulation of cytochrome c. These findings contribute to the understanding of Hsp27 anti‐apoptotic function and will aid in the development of anti‐apoptotic peptide therapies for the prevention of CVD in cancer patients.This research was funded by AHA SURF and ASC URS to JCT and RO1 HL075040 and NSF‐MCB 0542244 to AID
Genetic characterization of the mechanism of action of sulforaphane in the yeast, S. cerevisiae
Authors: Tucker Douglass , Murphy Michael , Wilcox Alexander , Austriaco Nicanor 
Published: June 2021

The isothiocyanate, sulforaphane (SFN), isolated from broccoli and other cruciferous vegetables is a potential chemotherapeutic agent. Recent studies that investigated the mechanism of action of SFN in mammalian cells have suggested that SFN works by causing cell cycle arrest and/or initiating programmed cell death (PCD). In order to identify genes that may be involved in the cell's response to SFN, we have initiated a genetic screen using the yeast knockout (YKO) library of the budding yeast, Saccharomyces cerevisiae, to identify loss of function mutants that are sensitive to SFN. At the time of this writing, we have screened the entire YKO collection twice, and we have isolated 54 mutants that were SFNS in both screenings of the library. We also have 174 single hits that need to be tested again. Significantly, we have discovered that mutations in several genes required for autophagy including VAM3 and VAM7 result in SFN sensitivity. This appears to confirm an earlier finding from our lab that yeast cells lacking ATG1, a kinase essential for autophagy, are also sensitive to sulforaphane, suggesting a possible role for autophagy in the mechanism of SFN‐induced programmed cell death in yeast.This research has been funded by a faculty research (CAFR) award from Providence College, a faculty development award from the Rhode Island‐INBRE program, and by grant R15 GM094712 from the NIGMS.
Ultrastructural analysis of experimental tenoplasty using bovine peritoneum
Authors: Masuko Telma Sumie 
Published: June 2021

Frequent lesions to tendons and ligaments bring lots of concern to orthopedists and veterinaries. Rupture of the calcaneal tendon is one of the most common tendon injury observed in the dog. The calcaneal tendon, also known as Achilles tendon or occasionally Achilles’ tendon is a tendon of the posterior leg and it is the strongest tendon in the structure of the musculoskeletal system in the dog. In dogs the calcaneal tendon originates from different muscle‐tendon units, the gastrocnemius tendon, the superficial digital flexor tendon, and the common tendon of the biceps femoris, gracilis and semitendinosus muscles. The aim of this study is to examine by scanning electron microscopy 98%‐glycerin‐fixed bovine peritoneum (GFP) used to fix lesions in dog's calcaneal tendon.Twenty one mixed bread healthy adult dogs, weighting between 10 and 15 kg were submitted to the implantation of a 1 cm‐GFP graft and sacrified 2, 7, 15, 30, 60, 90 and 120 days after the surgery. Fragments from 3 different segments, proximal, middle and distal, of the implanted graft were processed for the scanning electron microscopy analysis. The samples were fixed in 2% glutaraldehyde, post‐fixed in 1% Osmium Tetroxide, dehydrated in a series of alcohol, sputtered and observed in the scanning electron microscope.In all studied dogs, the peritoneum graft has been easily found without significant macroscopic derangements. Fifty five days after surgery, dogs were able to walk normally. The analysis under electron microscopy showed a presence of collagen tissue formation after 30 days post‐operative In this period we could observed the formation of neotendon involving peritoneum and nylon thread implanted in the segment. With 120 days post‐operative surgery can observe the presence of abundant collagen fibers arranged longitudinally, with morphology similar to the calcaneous tendon.Putting all into a nutshell, it is possible to conclude that glycerin‐fixed bovine peritoneum would be an interesting alternative for fixing tendon lesions as it seems to stimulate connective tissue deposition in order to form a good matrix for new tissue development.
Thrombospondin‐1 binding protein on the surface of Trypanosoma cruzi enhances cellular infection
Authors: Johnson Candice A. , Cardenas Tatiana C. , Pratap Siddharth , Duquette Mark A. , Lima Maria F. , Lawler Jack , Villalta Fernando , Nde Pius N. 
Published: June 2021

The mechanism by which Trypanosoma cruzi enhances cellular infection remains to be well elucidated. We have reported that T. cruzi up‐regulates the expression of host thrombospondin‐1 (TSP‐ 1) to enhance cellular invasion. Here we characterize a novel TSP‐ 1 interaction with T. cruzi that enhances cellular infection. Labeled TSP‐1 and the N‐terminal domain of TSP‐1 (NTSP) interact specifically with the surface of T. cruzi trypomastigotes. TSP‐1 and NTSP pulled down a novel TSP‐1‐interacting parasite surface protein in a protein‐protein interaction assay. We also observed that pre‐exposure of rNTSP to T. cruzi significantly enhances parasite infection of wild type mouse embryo fibroblast (MEF) in contrast to the C‐terminal domain of TSP‐1 (E3T3C1). Our findings indicate that TSP‐1 interacts with a novel protein on the surface of T. cruzi through the NTSP domain and that this amplifies the infection process.(Supported by NIH grants SC2AI083925 to NPN, 1 U54 RR026140‐03, AI007281‐23)
Zinc and copper status are associated with bone mineral measures in lactating adolescent mothers
Authors: Bezerra Flávia F. , Maia Patricia A. , Anastácio Alexandra , Mendonça Laura M.C. , Donangelo Carmen M. 
Published: June 2021

We investigated the relationship between bone mass and copper and zinc status in lactating (LA, n=33, 8.7±3.4wk postpartum) and non‐lactating (NL, n=14) adolescents (14–19y). Total bone mineral content (TBMC), total (TBMD) and lumbar spine (L2‐L4, LSBMD) bone mineral density, dietary intake, copper (CuP), zinc (ZnP) and ceruloplasmin (Cp) in plasma, and zinc (ZnE) and metallothionein (MT‐E) in erythrocytes were assessed. Over 70% of adolescents in each group had zinc and copper intakes lower than RDA. Biochemical indices were similar between groups except for ZnE and MT‐E that were 26% and 123% higher, respectively, in LA than in NL (P<0.05). LSBMD z score was lower in LA than in NL (−1.09 ± 1.5 vs. 0.04 ± 1.23, P<0.05). In LA, zinc intake was correlated with TBMC (r=0.38), TBMD z score (r=0.37) and LSBMD z score (r=0.37); ZnP was correlated with TBMC (r=0.44); and Cp with TBMD (r=0.50), LSBMD (r=0.42) and LSBMD z score (r=0.39) (P<0.05). In NL, no significant correlations between bone mass indices and zinc or copper status were observed. Our results suggest that a poor copper and zinc nutritional status may exacerbate the magnitude of bone loss during lactation in adolescent mothers.Financial support: CNPq, FAPERJ (Brazil).
Introduction of complementary feeding for infants: a dilemma for parents and caregivers
Authors: Friel James Kenneth , Qasem Wafaa , Fenton Tanis 
Published: June 2021

Data are scarce with respect to infant iron status, but at least in developing country settings where newborn iron stores may be suboptimal, exclusive breastfeeding without iron supplementation through 6 months may compromise hematologic status (WHO 2002). Our objective was to systematically review the scientific literature regarding iron and hematologic status among infants breastfed exclusively to 6 months, stratified by developed versus developing countries.MethodsAn electronic search of the MEDLINE database was conducted for studies of iron deficiency (ID) and (IDA) in infants exclusively breastfed in developed and developing countries. Prevalence estimates were calculated as weighted averages, and compared using a 2‐sided test of proportions.ResultsTwenty nine observational studies and three of eight RCTs met the inclusion criteria. All RCTs were conducted in developing countries. The prevalence estimates for healthy exclusively breastfed infants living in developed (n=769) and developing (n=642) countries, respectively, was 21.9% and 14.1% for ID (p = 0.0001), and 12.2% and 18.2% (p= 0.0019).ConclusionA substantial proportion of exclusively breastfed infants develop ID, IDA, and compromise hematologic status in both developed and developing countries. Data to support the current recommendation of solid commencement at six months is limited regarding irons status. Larger randomized controlled trials, perhaps in multi centre design, are needed to investigate the advantages and disadvantages of introduction of solids before and after six months of age.
The relationship between the use of blood pressure medication, a low‐sodium diet, and health and nutrition literacy among Community‐dwelling hypertensive Older Adults
Authors: Jones Samira , Young Heather , Miller Lisa 
Published: June 2021

More than 90 million U.S. adults have low health literacy, which is the ability to obtain and understand health information in order to make appropriate health decisions. Nutrition literacy is a component of health literacy pertaining to nutrition information. Nearly 68 million adults have hypertension, a preventable risk factor for cardiovascular disease and stroke; and persons with limited knowledge and understanding of diet and health concepts such as hypertension, generally have low health literacy and poor medication adherence. Community‐dwelling seniors in Sacramento County completed a modified health and nutrition knowledge survey, and two validated health literacy assessments: 1) the short Test for Functional Health Literacy (s‐ToFHLA) and 2) the Newest Vital Signs (NVS) to measure health or nutrition literacy. The goal was to determine the relationship between health and nutrition ˈknowledgeˈ and ˈliteracyˈ among hypertensive older adults (≥ 65 years). The sample included 69 older adults with an average age of 72 (SD= 7.3), and 26% African American, 9% Hispanic, and 58% White participants. Study findings included a modest correlation between the total literacy score and total knowledge score (r=0.58, p = 0.04), and significantly lower literacy scores for participants on blood pressure medication compared to those who were not (n=27) (95% CI: −83.04, −18.49%, p ≤ 0.01). No significant differences in scores were found for those on a low‐sodium diet compared to those who were not (p= 0.17). Future studies are needed to understand the relationship of health literacy to health outcomes to facilitate the development of tailored, effective interventions for older adults with chronic conditions.*Funding for this study was provided by the UC Davis Betty Irene Moore School of Nursing
Metabolic assessment of pediatric patients with chronic disease
Authors: Leung Randy B. , Schaub John R. , Klein Daniel S. , Borum Peggy R. 
Published: June 2021

BackgroundChronic disease can exacerbate obesity, type II diabetes, high blood pressure, and problems with children's growth. Changes in diet can impact these health issues.ObjectiveDevelop a program for a comprehensive metabolic assessment of patients seen in pediatric clinics for chronic disease.MethodsPatient interviews conducted at a weekly clinic for pediatric HIV+ patients included a 24‐hour diet recall; activity recall; height, weight, anthropometric skinfold and circumference measurements; and bioelectrical impedance analysis. Tools were created and refined to better define the above concerns, in order to better evaluate health issues.ResultsTools were developed for the following concerns and tested at a weekly clinic:

Assessment Concerns

Patient's MyPlate education

Patient's Solid Fats and Added Sugars intakes

Caloric requirements due to chronic disease

Quantitative analysis of medication load

Analysis of body growth and fat distribution

Risk for developing insulin resistance

Trends of abnormal blood lipids

Risk for hypertension

ConclusionThe program facilitates the assessment of potential comorbidities associated with nutrition in chronic disease and increases clinicians’ awareness of opportunities to improve patient care through education about lifestyle changes.
Bioavailability, glucose disposal, and anti‐inflammatory properties of grape products in high fat‐fed obese mice
Authors: Chuang Chia-Chi , Shen Wan , Xie Guoxiang , Jia Wei , McIntosh Michael 
Published: June 2021

The objective of this study was to determine which polyphenols in grape powder (GP) extract (GPE) were absorbed and the effect of GPE and GP on attenuating insulin resistance and inflammation in obese mice. We found that serum levels of quercetin 3‐glucoside (Q3G) and rutin were increased after 1 h post‐gavage of GPE (1 g/kg body weight) and decreased to baseline levels within 3–8 h. Next, C57BL/6J mice were fed a high fat diet supplemented for 18 wk with 0.02% GPE or 3% GP. Obese mice supplemented with GP, but not GPE, had better glucose tolerance acutely (e.g., at 5 wk), but not chronically (e.g., at 10 or 15 wk) compared to controls. Notably, obese mice supplemented with GP, but not GPE, had lower inflammatory cytokine (e.g., tumor necrosis factor alpha or TNFa) and chemokine (e.g., monocyte chemoattractant protein‐ 1 or MCP‐1) levels in circulation and mRNA levels of inflammatory‐related genes (e.g., TNFa, interleukin‐6, MCP‐1, interferon gamma‐induced protein‐10, F4/80, CD11c) in white adipose tissue compared to controls at 18 wk. Consistent with these data, Q3G and rutin decreased MCP‐1 gene expression in primary human adipocytes treated with TNFa. Overall, we demonstrate that several grape polyphenols (i.e., quercetin metabolites) are bioavailable and quercetin‐rich GP improves glucose tolerance acutely and attenuates markers of inflammation chronically in high fat‐fed obese mice.Grant Funding Source: NCARS02288 and UNCG127131