Poly-2-methyl-2-oxazoline-modified bioprosthetic heart valve leaflets have enhanced biocompatibility and resist structural degeneration.

Bioprosthetic heart valves (BHV) fabricated from glutaraldehyde-fixed heterograft tissue, such as bovine pericardium (BP), are widely used for treating heart valve disease, a group of disorders that affects millions. Structural valve degeneration (SVD) of BHV due to both calcification and the accumulation of advanced glycation end products (AGE) with associated serum proteins limits durability. We hypothesized that BP modified with poly-2-methyl-2-oxazoline (POZ) to inhibit protein entry would demonstrate reduced accumulation of AGE and serum proteins, mitigating SVD. In vitro studies of POZ-modified BP demonstrated reduced accumulation of serum albumin and AGE. BP-POZ in vitro maintained collagen microarchitecture per two-photon microscopy despite AGE incubation, and in cell culture studies was associated with no change in tumor necrosis factor-α after exposure to AGE and activated macrophages. Comparing POZ and polyethylene glycol (PEG)-modified BP in vitro, BP-POZ was minimally affected by oxidative conditions, whereas BP-PEG was susceptible to oxidative deterioration. In juvenile rat subdermal implants, BP-POZ demonstrated reduced AGE formation and serum albumin infiltration, while calcification was not inhibited. However, BP-POZ rat subdermal implants with ethanol pretreatment demonstrated inhibition of both AGE accumulation and calcification. Ex vivo laminar flow studies with human blood demonstrated BP-POZ enhanced thromboresistance with reduced white blood cell accumulation. We conclude that SVD associated with AGE and serum protein accumulation can be mitigated through POZ functionalization that both enhances biocompatibility and facilitates ethanol pretreatment inhibition of BP calcification.

Damaging de novo mutations diminish motor skills in children on the autism spectrum.

In individuals with autism spectrum disorder (ASD), de novo mutations have previously been shown to be significantly correlated with lower IQ but not with the core characteristics of ASD: deficits in social communication and interaction and restricted interests and repetitive patterns of behavior. We extend these findings by demonstrating in the Simons Simplex Collection that damaging de novo mutations in ASD individuals are also significantly and convincingly correlated with measures of impaired motor skills. This correlation is not explained by a correlation between IQ and motor skills. We find that IQ and motor skills are distinctly associated with damaging mutations and, in particular, that motor skills are a more sensitive indicator of mutational severity than is IQ, as judged by mutational type and target gene. We use this finding to propose a combined classification of phenotypic severity: mild (little impairment of either), moderate (impairment mainly to motor skills), and severe (impairment of both IQ and motor skills).

Identification of an AP1-ZFP36 Regulatory Network Associated with Breast Cancer Prognosis.

It has been established that ZFP36 (also known as Tristetraprolin or TTP) promotes mRNA degradation of proteins involved in inflammation, proliferation and tumor invasiveness. In mammary epithelial cells ZFP36 expression is induced by STAT5 activation during lactogenesis, while in breast cancer ZFP36 expression is associated with lower grade and better prognosis. Here, we show that the AP-1 transcription factor components, i.e. JUN, JUNB, FOS, FOSB, in addition to DUSP1, EGR1, NR4A1, IER2 and BTG2, behave as a conserved co-regulated group of genes whose expression is associated to ZFP36 in cancer cells. In fact, a significant down-modulation of this gene network is observed in breast, liver, lung, kidney, and thyroid carcinomas compared to their normal counterparts. In breast cancer, the normal-like and Luminal A, show the highest expression of the ZFP36 gene network among the other intrinsic subtypes and patients with low expression of these genes display poor prognosis. It is also proposed that AP-1 regulates ZFP36 expression through responsive elements detected in the promoter region of this gene. Culture assays show that AP-1 activity induces ZFP36 expression in mammary cells in response to prolactin (PRL) treatment thorough ERK1/2 activation. These results suggest that JUN, JUNB, FOS and FOSB are not only co-expressed, but would also play a relevant role in regulating ZFP36 expression in mammary epithelial cells.

Measuring shared variants in cohorts of discordant siblings with applications to autism.

We develop a method of analysis [affected to discordant sibling pairs (A2DS)] that tests if shared variants contribute to a disorder. Using a standard measure of genetic relation, test individuals are compared with a cohort of discordant sibling pairs (CDS) to derive a comparative similarity score. We ask if a test individual is more similar to an unrelated affected than to the unrelated unaffected sibling from the CDS and then, sum over such individuals and pairs. Statistical significance is judged by randomly permuting the affected status in the CDS. In the analysis of published genotype data from the Simons Simplex Collection (SSC) and the Autism Genetic Resource Exchange (AGRE) cohorts of children with autism spectrum disorder (ASD), we find strong statistical significance that the affected are more similar to the affected than to the unaffected of the CDS (P value ∼ 0.00001). Fathers in multiplex families have marginally greater similarity (P value = 0.02) to unrelated affected individuals. These results do not depend on ethnic matching or gender.

[Technologies for resection and reconstruction of mandibular lesions].

Restoring the bony structure of the face is a significant challenge. Due to the complex three dimensional anatomy, the presence of critical structures and the variety and uniqueness of each deficiency it is essential to form the optimal surgical plan and to execute it precisely. Development of computer aided manufacturing (CAD/CAM - computer assisted design / computer assisted manufacturing) and advances in manufacturing technologies and material science brought a whole array of options and tools for the purpose of planning and performing computer assisted surgery. CAD / CAM technology allow for various application: Manufacturing of custom made implants. The surgeon may choose which of the application to use in any specific case or combine all of these abilities according to the complexity of the case. Two cases presenting the applications of these technologies will be reviewed.

Facial reconstruction after pre maxillary resection in a bilateral cleft lip and palate patient - a case report Technologies for resection and reconstruction of mandibular lesions.

Loss of the premaxilla is a rare event while treating patients with bilateral cleft lip and palate. Its resection is no longer an acceptable method of treatment in modern practice. Nowadays, most cases of premaxillary loss are secondary to treatment and manipulation of the premaxilla, and not due to intentional resection. In the following case presentation we present the treatment of a seven years old boy, presented to our institution after being treated for bilateral cleft lip and palate by resection of the premaxilla. Through a series of operations including bone grafting, distraction of the maxilla, and cartilaginous graft we managed to reconstruct a proper facial habitus, a convex facial profile and nasal projection. Restoring the bony structure of the face is a significant challenge. Due to the complex three dimensional anatomy, the presence of critical structures and the variety and uniqueness of each deficiency it is essential to form the optimal surgical plan and to execute it precisely. Development of computer aided manufacturing (CAD/ CAM - computer assisted design / computer assisted manufacturing) and advances in manufacturing technologies and material science brought a whole array of options and tools for the purpose of planning and performing computer assisted surgery. CAD/CAM technology allow for various application: Manufacturing of a physical anatomical model, Using the 3D model for hardware pre-bending, Manufacturing of cutting jigs and intraoperative fixation templates, Manufacturing of custom made implants. The surgeon may choose which of the application to use in any specific case or combine all of these abilities according to the complexity of the case. Two cases presenting the applications of these technologies will be reviewed.

Rhomboid family gene expression profiling in breast normal tissue and tumor samples.

Rhomboid is an evolutionary conserved and functionally diversified group of proteins composed of proteolytically active and inactive members that are involved in the modulation of multiple biological processes such as epidermal growth factor receptor signaling pathway, endoplasmic reticulum-associated degradation, cell death, and proliferation. Recently, several human rhomboid genes have been associated with the development of chronic myeloid leukemia and pituitary, colorectal, ovarian, and breast cancers. In this study, we evaluated the mRNA and protein expression profiles of rhomboid genes in cancer cell lines and breast tissue/tumor samples. In silico analysis of publicly available gene expression datasets showed that different rhomboid genes are specifically expressed according to the breast cancer intrinsic subtypes. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis showed a significant RHBDD2 mRNA overexpression in advanced breast cancer compared with normal tissue samples (p = 0.012). In addition, we found that RHBDL2 and PARL mRNA expression was associated with a low/intermediate histologic tumor grade (p = 0.024 and p = 0.015, respectively). Immunohistochemistry analysis showed a significant increase of RHBDD2 protein expression in association with breast cancer samples negative for progesterone receptor (p = 0.015). Moreover, protein expression analysis corroborated the quantitative RT-PCR results, indicating that breast primary tumors belonging to patients with a more disseminated disease expressed significantly increased levels of RHBDD2 protein compared with less disseminated tumors (p = 0.01).

Dosage-dependent phenotypes in models of 16p11.2 lesions found in autism.

Recurrent copy number variations (CNVs) of human 16p11.2 have been associated with a variety of developmental/neurocognitive syndromes. In particular, deletion of 16p11.2 is found in patients with autism, developmental delay, and obesity. Patients with deletions or duplications have a wide range of clinical features, and siblings carrying the same deletion often have diverse symptoms. To study the consequence of 16p11.2 CNVs in a systematic manner, we used chromosome engineering to generate mice harboring deletion of the chromosomal region corresponding to 16p11.2, as well as mice harboring the reciprocal duplication. These 16p11.2 CNV models have dosage-dependent changes in gene expression, viability, brain architecture, and behavior. For each phenotype, the consequence of the deletion is more severe than that of the duplication. Of particular note is that half of the 16p11.2 deletion mice die postnatally; those that survive to adulthood are healthy and fertile, but have alterations in the hypothalamus and exhibit a "behavior trap" phenotype-a specific behavior characteristic of rodents with lateral hypothalamic and nigrostriatal lesions. These findings indicate that 16p11.2 CNVs cause brain and behavioral anomalies, providing insight into human neurodevelopmental disorders.

A systematic review of task-sharing interventions for substance use and substance use disorder in low- and middle-income countries.

INTRODUCTION: Substance use (SU) and substance use disorders (SUDs) are associated with adverse health and socio-economic consequences. Due to the shortage of specialist healthcare providers, people with SUDs in low- and middle-income countries (LMICs) have limited access to adequate treatment. Task-sharing with non-specialist health workers (NSHWs) has the potential to improve treatment accessibility for these individuals. This review synthesizes the evidence on the effectiveness of task-sharing interventions for SU and SUDs outcomes in LMICs. METHODS: PsycINFO, MEDLINE, EMBASE, Global Health and CENTRAL databases were searched to identify eligible studies. Quality assessment was conducted using the Cochrane risk of bias (RoB2) and Risk of Bias in Non-randomized Studies of Interventions (ROBINS-I) tool. A narrative synthesis was undertaken to analyze the data. RESULTS: Nineteen RCTs and two quasi-experimental studies met the eligibility criteria, and the majority had a low risk of bias rating. NSHW-delivered interventions significantly impact SU and SUDs outcomes, particularly in reducing alcohol and other substance use, cessation of smoking, and use of opioids. Multiple sessions delivered via face-to-face interactions was the most utilized method for intervention delivery. There were variations in terms of components of the intervention across studies; however, the most common intervention strategies used were a) personalized feedback, b) psychoeducation, c) motivational enhancement, d) problem-solving, and e) coping skills. CONCLUSION: Our review highlights the growing interests in leveraging NSHWs to provide interventions to people with SU and SUDs in LMICs where access to treatment is limited. However, additional research is necessary to explore the effectiveness of these interventions and identify the specific active components linked to enhancing treatment outcomes on a broader scale.

Evaluation of image quality and radiation dose in computed tomography urography following tube voltage optimisation.

INTRODUCTION: Computed tomography urography (CTU) comprehensively evaluates the urinary tract. However, the procedure is associated with a high radiation dose due to multiple scan series and therefore requires optimisation. The study performed CTU protocol optimisation based on a reduction in tube voltage (kV) using quality assurance (QA) phantom and clinical images and evaluated image quality and radiation dose. METHODS: The study was prospectively conducted on patients referred for CTU. The patients were grouped into A and B and were scanned with the standard protocol, a protocol used for the routine CTU at the CT centre before optimisation, and optimised protocol, a protocol with reduced kV respectively. The protocols were first tried on a quality assurance (QA) phantom before being applied to patients, and image quality was assessed based on signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR). In addition, the clinical images were assessed based on the visibility of the anatomical criteria for CT images by five observers with >5 years of experience. The data were analysed using both visual grading characteristic (VGC) curves and statistical package for social sciences (SPSS) version 22.0. RESULTS: The dose was significantly lower in the optimised protocol with a 10 % reduction in both volume computed tomography dose index and (CTDIvol) and dose length product (DLP) for the phantom images, and a 26 % reduction in CTDIvol and 28 % in DLP for the clinical images. However, there was no significant difference in image quality noted between the standard and optimised protocols based on the quantitative and qualitative image quality evaluation using both the QA phantom and clinical images. CONCLUSION: The findings revealed a significant dose reduction in the optimised protocol. Further, image quality in standard and optimised protocols did not differ significantly based on quantitative and qualitative methods. IMPLICATION FOR PRACTICE: kV optimisation in contrast-enhanced procedures provides dose reduction and should be encouraged in the medical imaging departments.

Pediatric Device Clinical Trials Activity: 1999-2022.

OBJECTIVES: This study assessed the state of pediatric medical device (PMD) development by comparing PMD clinical trials to pediatric trials evaluating drugs and biologics, from 1999 to 2022. METHODS: The site https://www.clinicaltrials.gov was used to identify and quantify both PMD clinical trials and pediatric trials for drugs and biologics. Clinical specialty was also assessed. The institutions included were the 7 children's hospitals primarily affiliated with the Food and Drug Administration (FDA) Pediatric Device Consortia (PDC) grant program between 2018 and 2023. For a national comparison, an additional search assessed PMD trials across all US medical institutions. RESULTS: A total of 243 PMD clinical trials were identified at the FDA-PDC institutions on the basis of the year of initiation; the average number of PMD trials initiated per year per institution was 1.5 from 1999 to 2022. However, PMD trials significantly increased during the period 2014 to 2022 compared with 1999 to 2013 (P < .001); the rate of initiation of drug and biologic pediatric trials demonstrated no significant differences between these time periods. A national survey of all institutions initiating PMD trials, and drugs and biologics trials, identified 1885 PMD trials out of a total 12 943. A comparable trend was noted in the national survey with initiation of PMD trials increasing significantly from 2014 to 2022 (P < .001), compared with 1999 to 2013, whereas the rate of initiation of drug and biologic trials during these periods did not demonstrate a significant change. CONCLUSIONS: Although pediatric clinical trial initiation for drugs and biologics remained stable from 1999 to 2022, the rate of new PMD trials significantly increased during the period 2014 to 2022 at FDA-PDC institutions and nationally.

Neutral mitochondrial heteroplasmy and the influence of aging.

The development and maintenance of mitochondrial heteroplasmy has important consequences for both health and heredity. Previous studies using pathogenic mutations have shown considerable variability between maternally related individuals and studies of several D-loop polymorphisms have suggested a relationship between heteroplasmy and somatic aging. To broadly explore the variation of human heteroplasmy and to clarify the dynamics of somatic heteroplasmy over the course of lifespan, we analyzed mitochondrial sequence variation across a range of ages. We utilized array-generated single-nucleotide polymorphism data that were well correlated with independent measures of heteroplasmy. Significant levels of heteroplasmy were identified at 0.24% of sites evaluated. By examining mother-child pairs, we found that heteroplasmy was inherited (30%) but could occur de novo in offspring or, conversely, be present in mothers but eliminated in their children (70%). Cumulatively, mitochondrial heteroplasmy across the genome increased significantly with advanced age (r = 0.224, P =8 × 10(-30)). Surprisingly, changes in heteroplasmy were not uniform with some sites demonstrating a loss of variation (increased homoplasmy) with aging. These data suggest that both mutation and selective pressure affect blood mitochondrial DNA sequence over the course of the human lifespan and reveal the unexpectedly dynamic nature of human heteroplasmy.

Sharing parental genomes by siblings concordant or discordant for autism.

Studying thousands of families, we find siblings concordant for autism share more of their parental genomes than expected by chance, and discordant siblings share less, consistent with a role of transmission in autism incidence. The excess sharing of the father is highly significant (p value of 0.0014), with less significance for the mother (p value of 0.31). To compare parental sharing, we adjust for differences in meiotic recombination to obtain a p value of 0.15 that they are shared equally. These observations are contrary to certain models in which the mother carries a greater load than the father. Nevertheless, we present models in which greater sharing of the father is observed even though the mother carries a greater load. More generally, our observations of sharing establish quantitative constraints that any complete genetic model of autism must satisfy, and our methods may be applicable to other complex disorders.

Decreased serotonin transporter activity in the mitral valve contributes to progression of degenerative mitral regurgitation.

Degenerative mitral valve (MV) regurgitation (MR) is a highly prevalent heart disease that requires surgery in severe cases. Here, we show that a decrease in the activity of the serotonin transporter (SERT) accelerates MV remodeling and progression to MR. Through studies of a population of patients with MR, we show that selective serotonin reuptake inhibitor (SSRI) use and SERT promoter polymorphism 5-HTTLPR LL genotype were associated with MV surgery at younger age. Functional characterization of 122 human MV samples, in conjunction with in vivo studies in SERT-/- mice and wild-type mice treated with the SSRI fluoxetine, showed that diminished SERT activity in MV interstitial cells (MVICs) contributed to the pathophysiology of MR through enhanced serotonin receptor (HTR) signaling. SERT activity was decreased in LL MVICs partially because of diminished membrane localization of SERT. In mice, fluoxetine treatment or SERT knockdown resulted in thickened MV leaflets. Similarly, silencing of SERT in normal human MVICs led to up-regulation of transforming growth factor ß1 (TGFß1) and collagen (COL1A1) in the presence of serotonin. In addition, treatment of MVICs with fluoxetine not only directly inhibited SERT activity but also decreased SERT expression and increased HTR2B expression. Fluoxetine treatment and LL genotype were also associated with increased COL1A1 expression in the presence of serotonin in MVICs, and these effects were attenuated by HTR2B inhibition. These results suggest that assessment of both 5-HTTLPR genotype and SERT-inhibiting treatments may be useful tools to risk-stratify patients with MV disease to estimate the likelihood of rapid disease progression.

RHBDD2: a 5-fluorouracil responsive gene overexpressed in the advanced stages of colorectal cancer.

In previous studies, we identified rhomboid domain containing 2 (RHBDD2) gene to be markedly overexpressed in breast cancer patients that developed recurrence of the disease. In this study, we evaluated for the first time RHBDD2 gene expression in colorectal cancer (CRC). Five public available DNA microarray studies were compiled in a homogeneous dataset of 906 colorectal samples. The statistical analysis of these data showed a significant increase of RHBDD2 expression in the advanced stages of CRC (p < 0.01). We validated these findings by immunohistochemistry on 130 colorectal tissue samples; RHBDD2 protein overexpression was also observed in the advanced stages of the disease (p < 0.001). In addition, we investigated RHBDD2 expression in response to the chemotherapy agent 5-fluorouracile (5FU). We detected a significant increase of RHBDD2 mRNA and protein after 5FU treatment (20-40 µM; p < 0.001). Overall, these results showed that RHBDD2 overexpression might play a role in colorectal cancer progression.

Analysis of image quality and radiation dose in routine adult brain helical and wide-volume computed tomography procedures.

OBJECTIVES: To evaluate and compare image quality and radiation dose between the helical and wide-volume scans to determine the protocol that provides a lower radiation dose without loss in image quality. METHODS: The study was prospectively conducted on consented adult patients that presented for routine brain CT. Image quality and radiation dose were compared between the helical and wide-volume scans on the Toshiba 160-slice Aquilion Prime CT scanner. The volume computed tomography dose index (CTDIvol) and dose length product (DLP) for each scan mode were collected and compared. Image quality was quantitatively and qualitatively evaluated using the unenhanced brain CT images. The data were analysed using a statistical package for social sciences (SPSS) software version 20 for both the descriptive and inferential statistics. A significant difference in image quality and radiation dose between the helical and wide-volume scans was determined based on a p-value of <0.05. RESULTS: A total of 54 participants were included, with two groups of 27 participants. The CTDIvol and DLP values were significantly p < 0.05 higher in the helical scan (CTDIvol: 65 mGy; DLP: 1597 mGy.cm) compared to the wide-volume scan (CTDIvol: 54 mGy; DLP: 1133 mGy.cm). The grey and white matters show a better signal-to-noise ratio (SNR) for the helical scan. Meanwhile, the contrast-to-noise ratio (CNR) was significantly p < 0.05 higher in the wide-volume scan. The results from the visual grading methods were compared and showed superior image quality in helical over the wide-volume scan. CONCLUSION: Wide-volume provides a lower dose compared to helical and therefore, could be adopted as the routine protocol for brain CT for in house dose optimisation. Where clinical conditions warrant the need for a helical scan, the protocol should be optimised in line with the as low as reasonable achievable (ALARA) principle.

Inhibition of advanced glycation end product formation and serum protein infiltration in bioprosthetic heart valve leaflets: Investigations of anti-glycation agents and anticalcification interactions with ethanol pretreatment.

Bioprosthetic heart valves (BHV) fabricated from heterograft tissue, such as glutaraldehyde pretreated bovine pericardium (BP), are the most frequently used heart valve replacements. BHV durability is limited by structural valve degeneration (SVD), mechanistically associated with calcification, advanced glycation end products (AGE), and serum protein infiltration. We investigated the hypothesis that anti-AGE agents, Aminoguanidine, Pyridoxamine [PYR], and N-Acetylcysteine could mitigate AGE-serum protein SVD mechanisms in vitro and in vivo, and that these agents could mitigate calcification or demonstrate anti-calcification interactions with BP pretreatment with ethanol. In vitro, each of these agents significantly inhibited AGE-serum protein infiltration in BP. However, in 28-day rat subdermal BP implants only orally administered PYR demonstrated significant inhibition of AGE and serum protein uptake. Furthermore, BP PYR preincubation of BP mitigated AGE-serum protein SVD mechanisms in vitro, and demonstrated mitigation of both AGE-serum protein uptake and reduced calcification in vivo in 28-day rat subdermal BP explants. Inhibition of BP calcification as well as inhibition of AGE-serum protein infiltration was observed in 28-day rat subdermal BP explants pretreated with ethanol followed by PYR preincubation. In conclusion, AGE-serum protein and calcification SVD pathophysiology are significantly mitigated by both PYR oral therapy and PYR and ethanol pretreatment of BP.

Early biometric lag in the prediction of small for gestational age neonates and preeclampsia.

OBJECTIVE: An early fetal growth lag may be a marker of future complications. We sought to determine the utility of early biometric variables in predicting adverse pregnancy outcomes. METHODS: In this retrospective cohort study, the crown-rump length at 11 to 14 weeks and the head circumference, biparietal diameter, abdominal circumference, femur length, humerus length, transverse cerebellar diameter, and estimated fetal weight at 18 to 24 weeks were converted to an estimated gestational age using published regression formulas. Sonographic fetal growth (difference between each biometric gestational age and the crown-rump length gestational age) minus expected fetal growth (number of days elapsed between the two scans) yielded the biometric growth lag. These lags were tested as predictors of small for gestational age (SGA) neonates (≤10th percentile) and preeclampsia. RESULTS: A total of 245 patients were included. Thirty-two (13.1%) delivered an SGA neonate, and 43 (17.6%) had the composite outcome. The head circumference, biparietal diameter, abdominal circumference, and estimated fetal weight lags were identified as significant predictors of SGA neonates after adjusted analyses (P < .05). The addition of either the estimated fetal weight or abdominal circumference lag to maternal characteristics alone significantly improved the performance of the predictive model, achieving areas under the curve of 0.72 and 0.74, respectively. No significant association was found between the biometric lag variables and the development of preeclampsia. CONCLUSIONS: Routinely available biometric data can be used to improve the prediction of adverse outcomes such as SGA. These biometric lags should be considered in efforts to develop screening algorithms for adverse outcomes.

Model studies of advanced glycation end product modification of heterograft biomaterials: The effects of in vitro glucose, glyoxal, and serum albumin on collagen structure and mechanical properties.

Glutaraldehyde cross-linked heterograft tissues, bovine pericardium (BP) or porcine aortic valves, are the leaflet materials in bioprosthetic heart valves (BHV) used in cardiac surgery for heart valve disease. BHV fail due to structural valve degeneration (SVD), often with calcification. Advanced glycation end products (AGE) are post-translational, non-enzymatic reaction products from sugars reducing proteins. AGE are present in SVD-BHV clinical explants and are not detectable in un-implanted BHV. Prior studies modeled BP-AGE formation in vitro with glyoxal, a glucose breakdown product, and serum albumin. However, glucose is the most abundant AGE precursor. Thus, the present studies investigated the hypothesis that BHV susceptibility to glucose related AGE, together with serum proteins, results in deterioration of collagen structure and mechanical properties. In vitro experiments studied AGE formation in BP and porcine collagen sponges (CS) comparing 14C-glucose and 14C-glyoxal with and without bovine serum albumin (BSA). Glucose incorporation occurred at a significantly lower level than glyoxal (p<0.02). BSA co-incubations demonstrated reduced glyoxal and glucose uptake by both BP and CS. BSA incubation caused a significant increase in BP mass, enhanced by glyoxal co-incubation. Two-photon microscopy of BP showed BSA induced disruption of collagen structure that was more severe with glucose or glyoxal co-incubation. Uniaxial testing of CS demonstrated that glucose or glyoxal together with BSA compared to controls, caused accelerated deterioration of viscoelastic relaxation, and increased stiffness over a 28-day time course. In conclusion, glucose, glyoxal and BSA uniquely contribute to AGE-mediated disruption of heterograft collagen structure and deterioration of mechanical properties.