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Heavy metals are ubiquitous in nature and their bioaccumulation usually remains undetected until it manifests as a chronic pathological condition. Accumulating evidence suggests that all age groups (independent of socioeconomic status, race, gender, and geographical location) and organs are susceptible to metal induced toxicity, although the impact on the central nervous system is more severe and long-lasting in both developing and adult human brain. There are no clinically proven treatments to those affected by heavy metal toxicity. We here review the most recent studies (2018–2020) on the differential impact of heavy metals on the pathogenesis of various neurodevelopmental disorders in children and neurodegenerative diseases in adults, highlighting the need for urgent intervention to this global health crisis.

Tissue microarchitecture and mechanics are important in development and pathologies of the Central Nervous System (CNS); however, their coordinating mechanisms are unclear. Here, we report that during colonization of the retina, microglia contacts the deep layer of high stiffness, which coincides with microglial bipolarization, reduction in TGFβ1 signaling and termination of vascular growth. Likewise, stiff substrates induce microglial bipolarization and diminish TGFβ1 expression in hydrogels. Both microglial bipolarization in vivo and the responses to stiff substrates in vitro require intracellular adaptor Kindlin3 but not microglial integrins. Lack of Kindlin3 causes high microglial contractility, dysregulation of ERK signaling, excessive TGFβ1 expression and abnormally-patterned vasculature with severe malformations in the area of photoreceptors. Both excessive TGFβ1 signaling and vascular defects caused by Kindlin3-deficient microglia are rescued by either microglial depletion or microglial knockout of TGFβ1 in vivo. This mechanism underlies an interplay between microglia, vascular patterning and tissue mechanics within the CNS., During development and pathologies of the central nervous system, blood vessels are closely connected with the innate immune cells, microglia. Here, the authors report that microglia adjust their angiogenic program to shape blood vessels and may depend on tissue stiffness.

Cell surface receptors are the key contributors of macrophage function. Most macrophage cell surface receptors are glycoproteins with sialic acids at the terminal of their glycans. It is well recognized that lipopolysaccharide (LPS) induces cell surface sialylation changes that may in turn contribute to macrophage functions. In addition, cellular mechanics such as elasticity is also a major determinant of macrophage function, which in turn is modulated by LPS. In this report, we characterized the sialylation status of macrophages upon LPS stimulation and assessed the changes in its mechanical properties and function. Specifically, we confirmed that sialylation status is closely related to macrophage biomechanical characteristics (elastic modulus, tether force, tether radius, adhesion force, and membrane tension) and thus directly involved in macrophage function. Further, we modulated macrophage sialylation status by feeding the cell with exogenous free sialic acid (Neu5Ac, Neu5Gc) and sialidase inhibitors, and examined the resulting effects on cellular mechanics and function. A systematic recognition of sialylation status related to cellular mechanics of macrophages will contribute to defining their phenotypes and elucidate macrophage functional diversity.

Controlling the free chlorine (FC) availability in wash water during sanitization of fresh produce enhances our ability to reduce microbial levels and prevent cross-contamination. However, maintaining an ideal concentration of FC that could prevent the risk of contamination within the wash system is still a technical challenge in the industry, indicating the need to better understand wash water chemistry dynamics. Using bench-scale experiments and modeling approaches, we developed a comprehensive mathematical model to predict the FC concentration during fresh-cut produce wash processes for different lettuce types (romaine, iceberg, green leaf, and red leaf), carrots, and green cabbage as well as Escherichia coli O157:H7 cross-contamination during fresh-cut iceberg lettuce washing. Fresh-cut produce exudates, as measured by chemical oxygen demand (COD) levels, appear to be the primary source of consumption of FC in wash water, with an apparent reaction rate ranging from 4.74×10-4to7.42×10-4 L/mg·min for all produce types tested, at stable pH levels (6.5 to 7.0) in the wash water. COD levels increased over time as more produce was washed and the lettuce type impacted the rate of increase in organic load. The model parameters from our experimental data were compared to those obtained from a pilot-plant scale study for lettuce, and similar reaction rate constant (5.38 × 10-4 L/mg·min) was noted, supporting our hypothesis that rise in COD is the main cause of consumption of FC levels in the wash water. We also identified that the bacterial transfer mechanism described by our model is robust relative to experimental scale and pathogen levels in the wash water. Finally, we proposed functions that quantify an upper bound on pathogen levels in the water and on cross-contaminated lettuce, indicating the maximum potential of water-mediated cross-contamination. Our model results could help indicate the limits of FC control to prevent cross-contamination during lettuce washing.

This study presents a modified disinfection kinetics model to evaluate the potential effect of organic content on the chlorine inactivation coefficient of Escherichia coli O157:H7 in fresh produce wash processes. Results show a significant decrease in the bactericidal efficacy of free chlorine (FC) in the presence of organic load compared to its absence. While the chlorine inactivation coefficient of Escherichia coli O157:H7 is 70.39 ± 3.19 L.mg−1.min−1 in the absence of organic content, it drops by 73% in chemical oxygen demand (COD) level of 600 - 800 mg.L−1. Results also indicate that the initial chlorine concentration and bacterial load have no effect on the chlorine inactivation coefficient. A second-order chemical reaction model for FC decay, which utilizes a percentage of COD as an indicator of organic content in fresh produce wash was employed, yielding an apparent reaction rate of (9.45 ± 0.22) × 10−4 μM−1.min−1. This model was validated by predicting FC concentration (R2 = 0.96) in multi-run continuous wash cycles with periodic replenishment of chlorine.

Elastogenesis is a complex process beginning with transcription, translation, and extracellular release of precursor proteins leading to crosslinking, deposition, and assembly of ubiquitous elastic fibers. While the biochemical pathways by which elastic fibers are assembled are known, the biophysical forces mediating the interactions between the constituent proteins are unknown. Using atomic force microscopy, we quantified the adhesive forces among the elastic fiber components, primarily between tropoelastin, elastin binding protein (EBP), fibrillin-1, fibulin-5, and lysyl oxidase-like 2 (LOXL2). The adhesive forces between tropoelastin and other tissue-derived proteins such as insoluble elastin, laminin, and type I collagens were also assessed. The adhesive forces between tropoelastin and laminin were strong (1767 ± 126 pN; p < 10−5vs. all others), followed by forces (≥200 pN) between tropoelastin and human collagen, mature elastin, or tropoelastin. The adhesive forces between tropoelastin and rat collagen, EBP, fibrillin-1, fibulin-5, and LOXL2 coated on fibrillin-1 were in the range of 100–200 pN. The forces between tropoelastin and LOXL2, LOXL2 and fibrillin-1, LOXL2 and fibulin-5, and fibrillin-1 and fibulin-5 were less than 100 pN. Introducing LOXL2 decreased the adhesive forces between the tropoelastin monomers by ∼100 pN. The retraction phase of force–deflection curves was fitted to the worm-like chain model to calculate the rigidity and flexibility of these proteins as they unfolded. The results provided insights into how each constituent's stretching under deformation contributes to structural and mechanical characteristics of these fibers and to elastic fiber assembly.

Background Single-chamber leadless pacemakers (LPs) have been shown to be an effective alternative to conventional transvenous pacemakers (CTPs), but their benefit in the context of cardioinhibitory vasovagal syncope (CI-VVS) is unknown. Objective The purpose of this study was to evaluate the safety and efficacy of LP compared with dual-chamber CTP for CI-VVS. Methods We conducted a multicenter, retrospective study comparing patients who received LP or dual-chamber CTP for drug-refractory CI-VVS. CI-VVS was diagnosed clinically and supported by cardiac monitoring and head-up tilt table testing. The primary efficacy endpoint was freedom from syncope during follow-up. Secondary endpoints included device efficacy and safety estimated by device-related major and minor adverse events (AEs). Results Seventy-two patients (24 LP, 48 CTP; age 32 ± 5.5 years; 90% female; syncope frequency 7.6 ± 3.4 per year) were included. At 1 year, 91% of patients (22/24) in the LP group and 94% of patients (43/48) in the CTP group met the primary efficacy endpoint (P = .7). Device efficacy endpoint was met in 92% of the LP group and 98% of the CTP group (P = .2). Early major AEs occurred in 2 of 24 in the LP group and 3 of 48 in the CTP group (P = .4). Late major AEs occurred in 0 of 24 in the LP group and 2 of 48 in the CTP group (P = 1). Conclusion In patients with CI-VVS, single-chamber LP demonstrated equivalent efficacy in reducing syncopal events compared to dual-chamber CTP, with a similar safety profile.

This study revealed that gut-derived n-butyrate protects against endometriosis by activating the expression of RAP1GAP GTPase to inhibit the RAP1 oncogenic pathway, indicating n-butyrate is linked with intestinal microbiota and endometriosis disease., Worldwide, ∼196 million are afflicted with endometriosis, a painful disease in which endometrial tissue implants and proliferates on abdominal peritoneal surfaces. Theories on the origin of endometriosis remained inconclusive. Whereas up to 90% of women experience retrograde menstruation, only 10% develop endometriosis, suggesting that factors that alter peritoneal environment might contribute to endometriosis. Herein, we report that whereas some gut bacteria promote endometriosis, others protect against endometriosis by fermenting fiber to produce short-chain fatty acids. Specifically, we found that altered gut microbiota drives endometriotic lesion growth and feces from mice with endometriosis contained less of short-chain fatty acid and n-butyrate than feces from mice without endometriosis. Treatment with n-butyrate reduced growth of both mouse endometriotic lesions and human endometriotic lesions in a pre-clinical mouse model. Mechanistic studies revealed that n-butyrate inhibited human endometriotic cell survival and lesion growth through G-protein–coupled receptors, histone deacetylases, and a GTPase activating protein, RAP1GAP. Our findings will enable future studies aimed at developing diagnostic tests, gut bacteria metabolites and treatment strategies, dietary supplements, n-butyrate analogs, or probiotics for endometriosis.

International audience; Abdominal aortic aneurysms (AAA) are characterized by matrix remodeling, elastin degradation, absence of nitric oxide (NO) signaling, and inflammation, influencing smooth muscle cell (SMC) phenotype and gene expression. Little is known about the biomolecular release and intrinsic biomechanics of human AAA-SMCs. NO delivery could be an attractive therapeutic strategy to restore lost functionality of AAA-SMCs by inhibiting inflammation and cell stiffening. We aim to establish the differences in phenotype and gene expression of adult human AAA-SMCs from healthy SMCs. Based on our previous study which showed benefits of optimal NO dosage delivered via S-Nitrosoglutathione (GSNO) to healthy aortic SMCs, we tested whether such benefits would occur in AAA-SMCs. The mRNA expression of three genes involved in matrix degradation (ACE, ADAMTS5 and ADAMTS8) was significantly downregulated in AAA-SMCs. Total protein and glycosaminoglycans synthesis were higher in AAA-SMCs than healthy-SMCs (p < 0.05 for AAA-vs. healthy-SMC cultures) and was enhanced by GSNO and 3D cultures (p < 0.05 for 3D vs. 2D cultures; p < 0.05 for GSNO vs. non-GSNO cases). Elastin gene expression, synthesis and deposition, desmosine crosslinker levels, and lysyl oxidase (LOX) functional activity were lower, while cell proliferation, iNOS, LOX and fibrillin-1 gene expressions were higher in AAA-SMCs (p < 0.05 between respective cases), with differential benefits from GSNO exposure. GSNO and 3D cultures reduced MMPs −2, −9, and increased TIMP-1 release in AAA-SMC cultures (p < 0.05 for GSNO vs. non-GSNO cultures). AAA-SMCs were inherently stiffer and had smoother surface than healthy SMCs (p < 0.01 in both cases), but GSNO reduced stiffness (~25%; p < 0.01) and increased roughness (p < 0.05) of both cell types. In conclusion, exogenously-delivered NO offers an attractive strategy by providing therapeutic benefits to AAA-SMCs.

RationaleAtrial fibrillation (AF) genome-wide association studies (GWAS) identified significant associations for rs1152591 and linked variants in the SYNE2 gene encoding the nesprin-2 protein that connects the nuclear membrane with the cytoskeletonObjectiveDetermine the effects of the AF-associated rs1152591 and rs1152595, two linked intronic single nucleotide polymorphisms (SNPs), on SYNE2 expression and investigate the mechanisms for their association with AF.Methods and ResultsRNA sequencing of human left atrial appendage (LAA) tissues indicated that rs1152591 and rs1152595 were significantly associated with the expressions of SYNE2α1, a short mRNA isoform, without an effect on the expression of the full-length SYNE2 mRNA. SYNE2α1 mRNA uses an alternative transcription start site and encodes an N-terminal deleted 62 kDa nesprin-2α1 isoform, which can act as a dominant-negative on nuclear-cytoskeleton connectivity. Western blot and qPCR assays confirmed that AF risk alleles of both SNPs were associated with lower expression of nesprin-2α1 in human LAA tissues. Reporter gene transfections demonstrated that the risk vs. reference alleles of rs1152591 and rs1152595 had decreased enhancer activity. SYNE2 siRNA knockdown (KD) or nesprin-2α1 overexpression studies in human stem cell-derived induced cardiomyocytes (iCMs) resulted in ~12.5 % increases in the nuclear area compared to controls (pConclusionsAF-associated SNPs rs1152591 and rs1152595 downregulate the expression of SYNE2α1, increasing nuclear-cytoskeletal connectivity and nuclear stiffness. The resulting increase in mechanical stress may play a role in the development of AF.

Transseptal puncture is a routinely performed interventional cardiology procedure for an array of cardiac diseases. We aimed to review the current status of available devices and techniques of transseptal puncture with consideration to specific interventions. Except for a few modifications, devices for transseptal puncture technique has not changed much compared to when it was first described almost 60 years ago. For difficult transseptal puncture, a few newer techniques such as radio frequency needle puncture system have been used but there is lack of robust clinical study. Advanced imaging, such as intracardiac echocardiography and transesophageal echocardiography, has been found to make transseptal puncture safer. A new transseptal approach that incorporates 3D non-fluoroscopic catheter tracking systems has shown promising results in two human studies. While various modifications in the transseptal technique tailored to the specific interventions have improved procedural safety, further improvement in existing devices focusing on distinct procedure might be needed in the future.

Regeneration following spinal cord injury (SCI) is challenging in part due to the modified tissue composition and organization of the resulting glial and fibrotic scar regions. Inhibitory cell types and biochemical cues present in the scar have received attention as therapeutic targets to promote regeneration. However, altered Young’s modulus of the scar as a readout for potential impeding factors for regeneration are not as well-defined, especially in vivo. Although the decreased Young’s modulus of surrounding tissue at acute stages post-injury is known, the causation and outcomes at chronic time points remain largely understudied and controversial, which motivates this work. This study assessed the glial and fibrotic scar tissue’s Young’s modulus and composition (scar morphometry, cell identity, extracellular matrix (ECM) makeup) that contribute to the tissue’s stiffness. The spatial Young’s modulus of a chronic (~18-wks, post-injury) hemi-section, including the glial and fibrotic regions, were significantly less than naïve tissue (~ 200 Pa; p < 0.0001). The chronic scar contained cystic cavities dispersed in areas of dense nuclei packing. Abundant CNS cell types such as astrocytes, oligodendrocytes, and neurons were dysregulated in the scar, while epithelial markers such as vimentin were upregulated. The key ECM components in the CNS, namely sulfated proteoglycans (sPGs), were significantly downregulated following injury with concomitant upregulation of unsulfated glycosaminoglycans (GAGs) and hyaluronic acid (HA), likely altering the foundational ECM network that contributes to tissue stiffness. Our results reveal the Young’s modulus of the chronic SCI scar as well as quantification of contributing elastic components that can provide a foundation for future study into their role in tissue repair and regeneration.

The assessment of neurotoxicity has been performed traditionally with animals. However, in vivo studies are highly expensive and time-consuming, and often do not correlate to human outcomes. Thus, there is a need for cost-effective, high-throughput, highly predictive alternative in vitro test methods based on early markers of mechanisms of toxicity. High-content imaging (HCI) assays performed on three-dimensionally (3D) cultured cells could provide better understanding of the mechanism of toxicity needed to predict neurotoxicity in humans. However, current 3D cell culture systems lack the throughput required for screening neurotoxicity against a large number of chemicals. Therefore, we have developed miniature 3D neural stem cell (NSC) culture on a unique 384-pillar plate, which is complementary to conventional 384-well plates. Mitochondrial membrane impairment, intracellular glutathione level, cell membrane integrity, DNA damage, and apoptosis have been tested against 3D-cultured ReNcell VM on the 384-pillar plate with four model compounds rotenone, 4-aminopyridine, digoxin, and topotecan. The HCI assays performed in 3D-cultured ReNcell VM on the 384-pillar plates were highly robust and reproducible as indicated by the average Z′ factor of 0.6 and CV values around 12%. From concentration-response curves and IC(50) values, mitochondrial membrane impairment appears to be the early stage marker of cell death by the compounds.

Human heart has endogenous regenerative capability; however, the intrinsic repair mechanism is not sufficient to overcome the impact placed by adverse pathological conditions, such as myocardial infarction (MI). In such circumstances, the damaged tissue initiates a series of remodeling process which results in the deterioration of structural, functional, and mechanical properties of the myocardium. To address such adverse conditions, clinical approaches ranging from surgical interventions, pharmaceutical drugs, and device implantation are administered which have played significant role in reducing the mortality rate. However, these approaches do not replace the lost cardiomyocytes, or restore the degraded structure-function relationship of the myocardium. In this aspect, cell-based therapy has gained substantial interest as a potential clinical approach for myocardial regeneration; however this method is impeded by lower graft retention and poor cell viability. To overcome these limitations, biomaterials are being developed as "trojan horses", i.e., vehicles for homing and deploying cells, and as matrices for delivering specific biological, mechanical, and chemical cues intended for tissue regeneration. Similarly, several candidate drugs, potent synthetic and biological molecules, and advanced drug delivery systems are being examined to provide exogenous cues in a controlled fashion to the diseased myocardium. In this article, we review biomaterials-based drug delivery systems for myocardial regeneration, specifically on the applications of hydrogels, microgels, nanoparticles, and nanofibers in the field. The prime focus of the article is on nanofibers-based drug delivery systems that is gaining considerable attention as a biomimetic pharmacological approach. We highlight literature on fabrication methods of self-assembling and electrospun nanofibers, drug incorporation methods and release kinetics, and in vitro and in vivo outcomes from nanofiber-based drug delivery systems in cardiac regeneration.

Endogenous adult cardiac regenerative machinery is not capable of replacing the lost cells following myocardial infarction, often leading to permanent alterations in structure-function-mechanical properties. Regenerative therapies based on delivering autologous stem cells within an appropriate 3D milieu could meet such demand, by enabling homing and directed differentiation of the transplanted cells into lost specialized cell populations. Since type I collagen is the predominant cardiac tissue matrix protein, we here optimized the 3D niche which could promote time-dependent evolution of cardiomyogenesis from human bone marrow-derived mesenchymal stem cells (BM-MSC). 3D collagen gel physical and mechanical characteristics were assessed using SEM and AFM, respectively, while the standalone and combined effects of collagen concentration, culture duration, and 5-azacytidine (aza) dose on the phenotype and genotype of MSC spheroids were quantified using immunofluorescence labeling and RT-PCR analysis. Increasing collagen concentration led to a significant increase in Young's modulus (p < 0.01) but simultaneous decrease in the mean pore size, resulting in stiffer gels. Spheroid formation significantly modulated MSC differentiation and genotype, mostly due to better cell-cell interactions. Among the aza dosages tested, 10 μM appears to be optimal, while 3 mg/ml gels resulted in significantly lower cell viability compared to 1 or 2 mg/ml gels. Stiffer gels (2 and 3 mg/ml) and exposure to 10 μM aza upregulated early and late cardiac marker expressions in a time-dependent fashion. On the other hand, cell-cell signaling within the MSC spheroids seem to have a strong role in influencing mature cardiac markers expression, since neither aza nor gel stiffness seem to significantly improve their expression. Western blot analysis suggested that canonical Wnt/β-catenin signaling pathway might be primarily mediating the observed benefits of aza on cardiac differentiation of MSC spheroids. In conclusion, 2 mg/ml collagen and 10 μM aza appears to offer optimal 3D microenvironment in terms of cell viability and time-dependent evolution of cardiomyogenesis from human BM-MSCs, with significant applications in cardiac tissue engineering and stem cell transplantation for regenerating lost cardiac tissue.

A full-term female neonate with ectrodactyly, polycystic kidney disease, mandibular hypoplasia, uterus unicornis, bronchogenic cyst and spina bifida is reported. The features are compared with other cases of limb and renal abnormalities reported in the literature. This case along with similar cases reported by Halal et al. (1980), Evans et al. (2000); Tobias et al. (2001); Phadke and Manisha (2006); John (2007); and Girisha et al. (2012) delineates acrorenal mandibular syndrome as a distinct entity.

A major onset of heart failure is myocardial infarction, which causes the myocardium to lose cardiomyocytes and transform into a scar tissue. Since mammalian infarcted cardiac tissue has a limited ability to regenerate, alternative strategies including implantation of tissue-engineered scaffolds at the site of damaged myocardium have been explored. The goal is to enable in situ cardiac reconstruction at the injured myocardium site, replace the lost cardiomyocytes, deliver the required biomolecules, and remodel the extracellular matrix (ECM). ECM synthesis and deposition by cardiomyocytes within such scaffolds remains categorically unexplored. Here, we investigated the survival, ECM synthesis and deposition, and matrix metalloproteinases (MMPs) release by cardiomyocytes within three-dimensional (3D) substrates. Rat cardiomyocytes were cultured for three weeks within two structurally different substrates: 3D collagen hydrogels or polycaprolactone (PCL) nanofibrous scaffolds. The concentration and composition of the hydrogels was varied, while PCL nanofibers were surface-modified with various ECM proteins. Results showed that myocyte attachment and survival was higher within collagen hydrogels, while myocyte alignment and beating was noted only within PCL scaffolds. Total protein synthesis by myocytes within PCL scaffolds was significantly higher compared to that within collagen hydrogels, although more protein was deposited as matrix within hydrogels. Significant ECM synthesis and matrix deposition, TIMP-1, and MMP release were noted within modified collagen hydrogels and PCL nanofiber scaffolds. These results were qualitatively confirmed by imaging techniques. Results attest to the prominent role of scaffold composition and architecture in influencing cardiomyocyte phenotype, matrix synthesis and cytokines release, with significant applications in cardiac tissue remodeling strategies.

Under disease or injury conditions in the central nervous system (CNS), activated microglia release cytokines and chemokines to modulate the microenvironment and influence tissue remodeling. To exploit the full potential of neural stem cell (NSC) transplantation approaches, a permissive microenvironment needs to be created for their survival, homing and differentiation. To investigate the role of chronically activated microglia in the fate of NSCs, spontaneously immortalized murine microglial cells (SIM-A9) were cocultured with embryonic murine cortical NSCs on 2D substrates or within 3D gels. Standalone NSC cultures served as controls. Cytokines and chemokines released by NSCs and SIM-A9 cells in standalone and cocultures were quantified. Coculturing with SIM-A9 cells suppressed NSC viability, neurite outgrowth, neural differentiation and TUJ1 gene expression, and promoted glia formation in both 2D and 3D cultures, over a 10-day period. The seven most-abundantly released analytes by microglia (MCP-1, MIP2, G-CSF, MIP-1α, MIP-1β, TNF-α, IL-6) were tested for their individual effects on NSCs, to investigate if the outcomes in cocultures were due to the synergistic effects of analytes or the influence of any individual analyte. All the seven analytes significantly suppressed cell survival compared to controls, but exposure to MIP-1β, IL-6, or MCP-1 enhanced neurite outgrowth and neural lineage commitment. Results attest to (i) the strong role of activated microglia in regulating NSC fate, (ii) the utility of selective analytes released by activated microglia in promoting neurogenesis and neuritogenesis, and (iii) the need to protect transplanted NSCs from the host inflammatory microenvironment to ensure their survival and functionality in treating neurological disorders.

Exposure to environmental toxicants such as heavy metals, pesticides, and nanoparticles poses a severe threat to both the developing and the adult human brain, causing various neurodegenerative disorders. Detection and quantification of neurotoxicity induced by such toxicants represent a major challenge due to the complexity of neuronal pathways involved and a lack of understanding of mechanistic actions of toxicants in vivo. While the role of neural stem cells (NSCs) in this process is becoming increasingly evident, outcomes from conventional in vitro assays explored thus far are curtailed by the relative high-cost and low throughput (number of bioassays per time), physiologically-irrelevant 2D cell cultures, and unavailability of the requisite cell populations. This chapter aims to highlight the various mechanisms involved in neurotoxicity and neuronal differentiation of NSCs, and summarizes various in vitro assays currently being used for the detection of neurotoxicity along with their limitations.

Polygonum chinensis linn belongs to the family Polygonaceae. The plant used in this study was shade dried and extracted with methanol by a cold maceration process. Qualitiave phytochemicals estimation showed the presence of the majority of compounds including terpinoids, flavonoids, tannins, saponins and glycosides. All the extracts showed moderate inhibition ranged from 12- 14 mm against both Gram-positive and Gram-negative strains and 00-21 mm against fungal trains. In the two-fold serial dilution method, the minimum inhibitory concentration of the extract ranges between 250 – >1000 µg/ml. The leaf extract of polygonum chinensis showed the potent activity with an IC50 value of 18.59 ± 0.356 µg/ml. In DPPH, the lipid peroxidation and hydrogen peroxide method, the leaf extract showed potent antioxidant activity. Root and stem extract showed moderate antioxidant activity. All the extracts showed moderate cytotoxicity to all the five cell lines Vero, A-549, L6, BRL, 3A and L-929 with CTC50 value ranging from 400-125 µg/ml. Short-term cytotoxicity studies were conducted for all the extracts against (Dalton Lymphocytes ascites) cells. The CTC50 value was found to be 435 µg/ml. Thus, the extracts of P. Chinensis may possess cytotoxic, antioxidant and antimicrobial activity.

Elastin breakdown in vascular aneurysms is mediated by cytokines such as tumor necrosis factor alpha (TNF-alpha, which induces vascular smooth muscle cell (SMC) activation and regulates their deposition of matrix. We previously demonstrated that exogenous supplementation with TGF-beta1 (1 ng ml(-1)) and hyaluronan oligomers (0.786 kDa, 0.2 microg ml(-1)) cues the upregulation of elastin matrix synthesis by healthy cultured SMCs. Here, we determine whether these cues likewise enhance elastin matrix synthesis and assembly by TNF-alpha-stimulated SMCs, while restoring their healthy phenotype. Adult rat aortic SMCs were treated with TNF-alpha alone or together with TGF-beta1/hyaluronan oligomeric cues and the release of inflammatory markers were monitored during over a 21 day culture. Biochemical analysis was used to quantify cell proliferation, matrix protein synthesis and cross-linking efficiency, while immunofluorescence and electron microscopy were used to analyze the elastin matrix quality. It was observed that SMC activation with TNF-alpha (10 ng ml(-1)) induced matrix calcification and promoted production of elastolytic MMP-2 and MMP-9. However, these effects were attenuated by the addition of TGF-beta1 and HA oligomer cues to TNF-alpha-stimulated cultures, which also enhanced tropoelastin and collagen production, improved elastin matrix yield and cross-linking, promoted elastin fiber formation and suppressed elastase activity, although the release of MMP-2 and MMP-9 was not affected. Overall, the results suggest that TGF-beta1 and HA oligomers are potentially useful in suppressing SMC activation and inducing regenerative elastin repair within aneurysms.

The progression of aortic aneurysms (AAs) is typically associated with an activated smooth muscle cell (SMC) phenotype, diminished density of mature medial elastic fibers, and an elevated presence of matrix-degrading enzymes, which ultimately leads to vessel rupture. Currently, no surgical or nonsurgical methods are available to regress aneurysms via regeneration of new elastic matrices, particularly because of inherently poor elastin synthesis by adult vascular cells and absence of tools to stimulate the same. We seek to address this void in this study. We recently showed 0.2 microg/mL of hyaluronan oligomers and 1 ng/mL of transforming growth factor-beta1 (termed elastogenic factors) to dramatically enhance elastin synthesis and matrix formation by healthy aortic SMCs. In this study, the effect of these factors, alone or together, on suppressing procalcific and elastolytic activities of aneurysmal vascular cells, and improving their elastin matrix synthesis and assembly is examined. Periadventitial injury with calcium chloride was used to induce AAs in rats, and approximately 45% increase in aortic diameter was observed after 4 weeks. Aneurysmal SMCs isolated from these AA segments produced higher levels of inflammatory markers matrix metalloproteinases-2 and 9 elastase activity and calcific deposits, while synthesizing significantly less collagen, tropoelastin, and matrix elastin proteins over a 3-week culture period, relative to healthy SMCs. While hyaluronan oligomers alone significantly suppressed aneurysmal cell proliferation and promoted 20-50% increases in collagen and elastin synthesis (p0.01), transforming growth factor-beta1 alone had no effect on cellular proliferation and elastin synthesis. However, provision of factors together resulted in significantly higher amounts of collagen/elastin protein synthesis and crosslinking, by upregulating lysyl oxidase and desmosine. Compared to their individual contributions, the factors together were highly effective in minimizing the release of inflammatory enzymes, and encouraging elastic fiber formation. Since elastic matrix amounts were one order of magnitude lower than that observed with healthy cells, even upon elastogenic stimulation at doses optimized previously for healthy cells, increased doses are likely required and must be reoptimized for diseased cells. Despite this, the results point to the potential utility of these elastogenic factors in regenerating elastic matrices within AAs.

Introduction: The goals of this study were to (i) evaluate the feasibility and results of monitoring patients prior to and following catheter ablation of AF with the mobile cardiac outpatient telemetry (MCOT) system and to (ii) correlate symptoms and the presence or absence of atrial fibrillation (AF). Methods and Results: A total of 19 consecutive patients (12 men [63%]; age 60 ± 6 years) with highly symptomatic drug refractory AF underwent catheter ablation. Each was provided with a MCOT monitor (CardioNet, USA) and asked to wear it 5 days immediately before the ablation, and 5 days per month starting with the ablation for 6 consecutive months. When patients experienced any symptoms, they were asked to activate the system and to record associated symptoms. A total of 494 days (11,856 hours) were monitored. Out of the total 390 events triggered by patient's symptoms, 40% were confirmed as AF events (156) and 60% were confirmed as non-AF events (234). Only shortness of breath and chest discomfort were highly associated with AF (P < 0.05). At the end of 6 months of follow-up, out of 10 patients that completed the study, 7 (70%) patients were free of symptomatic AF recurrences whereas only 5 (50%) patients achieved success when asymptomatic AF recurrences were included in the outcome. Conclusions: The results of this study demonstrate the potential utility of wireless monitoring systems in the follow-up of AF ablation patients, while also identifying poor patient compliance with an intensive monitoring protocol as an important limitation.

Catheter ablation to achieve pulmonary vein (PV) isolation has become an increasingly used treatment strategy for patients with atrial fibrillation (AF). The purpose of this study was to evaluate the impact of segmental isolation of PVs on volume of left atrium and its relation to the decrease in the size of the pulmonary veins.Gadolinium enhanced Magnetic Resonance Angiography (MRA) was performed in 51 AF patients before and 6 approximately 8 weeks post PV isolation, using cooled radio-frequency (RF) energy. Three-dimensional reconstruction with maximum intensity projections and multiplanar reformations was performed. Oblique coronal projections were used to measure the ostial size of PVs. Three orthogonal dimensions of LA chamber were measured and computed to assess the volume of the left atrium.The mean LA volume decreased by 15.7% after ablation (p0.001). The mean PV ostial diameter decreased by 11%, from 18.3+/-0.8 mm to 16.7+/-1.0 mm (p=0.005). Moderate PV stenosis was noted in two veins out of the 192 veins analyzed. There was a significant correlation between changes in the size of PV ostium to that of the LA.Catheter ablation of AF using a segmental PV isolation approach results in a significant reverse remodeling in the left atrium. Significant stenosis of PVs appears to be rare after the segmental isolation procedure.

Objectives The purpose of this study was to report the safety, efficacy, and predictors of recurrence of circumferential pulmonary vein (PV) catheter ablation in patients with atrial fibrillation (AF). Background Circumferential PV ablation has been described as an alternate ablation strategy for AF. Methods Seventy consecutive patients (age 56 ± 10 years) with symptomatic drug refractory paroxysmal (n = 21), persistent (n = 22), and permanent (n = 27) AF underwent catheter ablation. The catheter ablation procedure was performed by creating circular lesions encircling right- and left-side PV ostia guided by an electroanatomic (CARTO) mapping system. Linear ablation lesions also were created in the cavotricuspid isthmus, the mitral isthmus, and in the posterior left atrium. In 42 patients (60%), additions linear lesions were created between superior and inferior PVs in a "figure-of-eight" fashion. Results At 6 ± 2.5 months of follow-up, 53 patients (76%) were AF free, including 39 patients (56%) not taking and 14 patients (20%) taking antiarrhythmic drugs. Among various variables, only early recurrence of AF was a predictor of long-term recurrence. Significant complications included one pericardial tamponade, one stroke, and two PV occlusions. Both patients with PV occlusion received radiofrequency delivery in a figure-of-eight fashion. Conclusions Circumferential PV catheter ablation of AF is associated with moderate efficacy and risk of complications. The absence of a difference in efficacy combined with the risk of PV stenosis associated with figure-of-eight lesion lead us to conclude that the figure-of-eight lesion should not be a routine component of circumferential PV AF ablation procedures.

Metabolic syndrome is a cluster of risk factors for cardiovascular disease that include obesity, atherogenic dyslipidemia, raised blood pressure, and insulin resistance. The growing trend of obesity is associated with increased prevalence of metabolic syndrome. Optimizing diet and exercise are still the leading therapy for controlling the metabolic syndrome. Based on the current evidence, further emphasis should be placed on aggressive management of other metabolic risk factors such as high blood pressure and dyslipidemia.

Favorable results of circumferential pulmonary vein (PV) ostial ablation guided by electroanatomic mapping (Carto) have fueled great enthusiasm for this technique. The lesion set for this ablation procedure as described by Pappone et al.(1,2) involves a figure of 8 lesions encompassing the 2 right and 2 left PV ostia with 2 additional linear lesions. Pulmonary vein stenosis/or occlusion has not previously been reported as a complication of this procedure. We describe PV occlusion after this procedure. Based on the occurrence of this previously unreported complication, we no longer create a figure '8' lesion around the right superior and inferior PVs. We anticipate that this complication can be avoided by creating a circular lesion encompassing the 2 PV ostia, while avoiding the linear lesion between the 2 PV ostia.

Introduction: Delineation of pulmonary vein (PV) anatomy is an integral part of the PV isolation procedure. The aims of the present study were to (1) describe the technique of selective PV angiography, (2) show the typical fluoroscopic locations and appearance of the PVs, and (3) compare the ostial diameters of PVs measured by angiography and magnetic resonance imaging (MRI). Methods and Results: Twenty consecutive patients undergoing a PV isolation procedure underwent selective PV angiography using a deflectable 8-French lumened catheter (Naviport, Cardima). The left superior PV (LSPV) runs upward and away from the spine in the right anterior oblique (RAO) projection and upward and toward the spine in the left anterior oblique (LAO) projection. The opposite is true for the right superior PV (RSPV). The left inferior PV (LIPV) has a bull's-eye appearance in the RAO projection, and the right inferior PV (RIPV) has a bull's-eye appearance in the LAO projection due to their end-on trajectories. The LIPV in the LAO projection and the RIPV in the RAO projection run horizontally toward the spine. An excellent correlation was noted in PV ostial size as assessed by angiography and MRI (r2 < 0.90, P < 0.0001). Conclusion: This study describes the technique and results of PV angiography and fluoroscopy. The study also demonstrates good correlation of PV ostial diameters by contrast venography and MRI. PV angiography can be used as an alternate to MRI or computed tomographic imaging, particularly when these tests are unavailable or are contraindicated in the patient. (J Cardiovasc Electrophysiol, Vol. 15, pp. 21-26, January 2004)

Studying neurite guidance by diffusible or substrate bound gradients is challenging with current techniques. In this study, we present the design, fabrication and utility of a microfluidic device to study neurite guidance under chemogradients. Experimental and computational studies demonstrated the establishment of a steep gradient of guidance cue within 30 min and stable for up to 48 h. The gradient was found to be insensitive to external perturbations such as media change and movement of device. The effects of netrin-1 (0.1–10 µg mL−1) and brain pulp (0.1 µL mL−1) were evaluated for their chemoattractive potential on neurite turning, while slit-2 (62.5 or 250 ng mL−1) was studied for its chemorepellant properties. Hippocampal or dorsal root ganglion (DRG) neurons were seeded into a micro-channel and packed onto the surface of a 3D collagen gel. Neurites grew into the matrix in three dimensions, and a gradient of guidance cue was created orthogonal to the direction of neurite growth to impact guidance. The average turning angle of each neurite was measured and averaged across multiple devices cultured under similar conditions to quantify the effect of guidance cue gradient. Significant positive turning towards gradient was measured in the presence of brain pulp and netrin-1 (1 µg mL−1), relative to control cultures which received no external guidance cue (p < 0.001). Netrin-1 released from transfected fibroblasts had the most positive turning effect of all the chemoattractive cues tested (p < 0.001). Slit-2 exhibited strong chemorepellant characteristics on both hippocampal and DRG neurite guidance at 250 ng mL−1 concentration. Slit-2 also showed similar behavior on DRG neuron invasion into 3D collagen gel (p < 0.01 relative to control cultures). Taken together, the results suggest the utility of this microfluidic device to generate stable chemogradients for studying neurobiology, cell migration and proliferation, matrix remodeling and co-cultures with other cell lines, with potential applications in cancer biology, tissue engineering and regenerative medicine., Seoul R&BD Support Center (program PA090930)

Persistent left superior vena cava (PLSVC) is the most common central venous anomaly in the thorax, which may remain asymptomatic throughout life. Indwelling catheters that are increasingly used in oncology practice may result in complications in the presence of PLSVC. The authors report an unusual case of recurrent ischemic attacks each time a portacath was accessed in a patient with breast cancer and PLSVC. This case highlights the importance of suspecting this condition in the presence of clinical and radiographic clues to prevent complications.

Tissue Engineering Approaches for Motor Neuron Pathway Regeneration During fetal development, a tightly-regulated spatio-temporal pattern of guidance cues directs and maintains motor neuron axonal growth along specific pathways to reach the target cells and tissues. However, an inflammatory environment resulting from an injury (e.g., trauma, stroke) or disease [e.g., amyotrophic lateral sclerosis (ALS), primary lateral sclerosis (PLS), and hereditary spastic paraplegia (HSP)] leads to progressive degeneration of motor neurons and destruction of axonal tracts in the adult CNS.

Microfluidic devices allow for precise control of the cellular and noncellular microenvironment at physiologically relevant length- and time-scales. These devices have been shown to mimic the complex in vivo microenvironment better than conventional in vitro assays, and allow real-time monitoring of homotypic or heterotypic cellular interactions. Microfluidic culture platforms enable new assay designs for culturing multiple different cell populations and∕or tissue specimens under controlled user-defined conditions. Applications include fundamental studies of cell population behaviors, high-throughput drug screening, and tissue engineering. In this review, we summarize recent developments in this field along with studies of heterotypic cell-cell interactions and tissue specimen culture in microfluidic devices from our own laboratory.

Health-related quality of life (HRQOL) is an important measure of the effects of chronic liver disease in affected patients that helps guide interventions to improve well-being. However, the relationship between HRQOL and survival in liver transplant candidates remains unclear. We examined whether the Physical Component Summary (PCS) and Mental Component Summary (MCS) scores from the Short Form 36 (SF-36) Health Survey were associated with survival in liver transplant candidates. We administered the SF-36 questionnaire (version 2.0) to patients in the Pulmonary Vascular Complications of Liver Disease study, a multicenter prospective cohort of patients evaluated for liver transplantation in 7 academic centers in the United States between 2003 and 2006. Cox proportional hazards models were used with death as the primary outcome and adjustment for liver transplantation as a time-varying covariate. The mean age of the 252 participants was 54 +/- 10 years, 64% were male, and 94% were white. During the 422 person years of follow-up, 147 patients (58%) were listed, 75 patients (30%) underwent transplantation, 49 patients (19%) died, and 3 patients were lost to follow-up. Lower baseline PCS scores were associated with an increased mortality rate despite adjustments for age, gender, Model for End-Stage Liver Disease score, and liver transplantation (P for the trend = 0.0001). The MCS score was not associated with mortality (P for the trend = 0.53). In conclusion, PCS significantly predicts survival in liver transplant candidates, and interventions directed toward improving the physical status may be helpful in improving outcomes in liver transplant candidates.

Elastin, a major component of vascular matrices, critically determines vascular mechanics and maintains the quiescence of smooth muscle cells (SMCs). Attempts to regenerate elastin in elastin-compromised blood vessels using tissue-engineering approaches is limited by the unavailability of elastogenic cues to upregulate poor elastin output and matrix assembly by adult vascular cells. We previously showed that hyaluronan (HA) elastogenically stimulates aortic SMCs, although these effects are highly specific to HA fragment size. The elastogenic response of SMCs can also be modulated with growth factors such as insulin-like growth factor (IGF-1). Here, we evaluate the benefits of concurrent delivery of HA fragments (0.76–2000 kDa) and IGF-1 (500 ng/ml) to elastin synthesis, organization and crosslinking. The study outcomes show that, relative to supplement-free cultures, IGF-1 and long-chain HA/large HA fragments, but not HA oligomers, together induce multifold increases in the synthesis of elastin precursors, structural elastin matrix yields and crosslink densities within cell layers, and encourage elastic fibre formation. These outcomes are not all obtained when either of the cues is provided separately. IGF-1 and large HA fragments (>20 kDa) also together inhibit cell proliferation, a concern in elastin-compromised vessels, where SMC hyperproliferation is common. The results will benefit efforts to provide exogenous or scaffold-based elastogenic cues (IGF-1 + HMW HA/large HA fragments) to enable robust and faithful regeneration of elastin matrix structures in vivo or in vitro. The present outcomes may be used to restore elastin matrix homeostasis in de-elasticized vessels and tissue-engineered constructs that may be grafted as a substitute. Copyright © 2008 John Wiley & Sons, Ltd.

Purpose: To develop an ibuprofen ointment with a potential for both topical and systemic delivery of the drug. Method: A co-solvency technique with a trial and error approach was used to develop a 10% ibioprofen ointment in petrolatum base, with the entire drug dissolved in the base. An insertion cell was used to evaluate drug release from the formulations. Further, factorial design multiple regression (FDMRA) analysis, a statistical optimization technique, was used in the optimization of the final formulation. Result: The desired ibuprofen ointments were developed. Release depended on vehicle and proportion of co-solvents. Best fit equations for optimization purposes including various fluxes (initial, steady-state and total) and diffusion coefficient as dependent variables and the concentrations of co-solvents as independent variables were obtained using SAS programme. Dependent variables strongly depended (p

Two important limitations of the data regarding the outcomes of catheter ablation of atrial fibrillation (AF) are the short-term follow-up used in most published studies and the lack of single-procedure outcomes. The objective was to report the long-term single-procedure outcomes at our center. The patient population was comprised of 200 consecutive patients who underwent ablation (133 men; age 56 ± 11 years). Atrial fibrillation was paroxysmal in 92 (46%). Success was defined as absence of symptomatic AF, off antiarrhythmic drug (AAD) after a single procedure. After a follow-up of 26 ± 11 months, the single-procedure long-term success rate was 28% with an additional 7% of patients demonstrating improvement. After including repeat procedures in 64 patients, the overall long-term success rate was 41% with 11% demonstrating improvement. Further subgroup analysis of 48 paroxysmal AF patients considered to be optimal candidates for the procedure, revealed a long-term success rate of 69% with an additional 4% demonstrating improvement. A major complication occurred in 7.9% of patients. The results reveal that the long-term single-procedure success rate of catheter ablation of AF in a cohort of patients with predominantly non-paroxysmal AF is less than 40%. The inclusion of redo procedures resulted in an improvement in outcomes. A much higher success rate of 69% was achieved in patients with paroxysmal AF considered to be optimal candidates for this procedure. These results make it clear that further advances in the technique of catheter ablation of AF are needed to improve the safety and efficacy of this procedure. In order to be able to compare outcomes of various techniques in differing patient populations, we urge investigators to report long-term single procedure outcomes.

Pulmonary Vein Stenosis After Atrial Fibrillation Ablation. introduction: There are currently no studies systematically evaluating pulmonary vein (PV) stenosis following catheter ablation of atrial fibrillation (AF) usingthe anatomic PV ablation approach. Methods and Results: Forty-one patients with AF underwent anatomic PV ablation under the guidance of a three-dimensional electroanatomic mapping system. Gadolinium-enhanced magnetic resonance (MR) imaging was performed in all patients prior to and 8-10 weeks after ablation procedures for screening of PV stenosis. A PV stenosis was defined as a detectable (≥3 mm) narrowing in PV diameter. The severity of stenosis was categorized as mild ( 70%). A total 157 PVs were analyzed. A detectable PV narrowing was observed in 60 of 157 PVs (38%). The severity of stenosis was mild in 54 PVs (34%), moderate in five PVs (3.2%), and severe in one PV (0.6%). All mild PV stenoses displayed a concentric pattern. Moderate or severe PV stenosis was only observed in patients with an individual encircling lesion set. Multivariable analysis identified individual encircling lesion set and larger PV size as the independent predictors of detectable PV narrowing. All patients with PV stenosis were asymptomatic and none required treatment. Conclusions: The results of this study demonstrate that detectable PV narrowing occurs in 38% of PVs following anatomic PV ablation. Moderate or severe PV stenosis occurs in 3.8% of PVs. The high incidence of mild stenosis likely reflects reverse remodeling rather than pathological PV stenosis. The probability of moderate or severe PV stenosis appears to be related to creation of individual encircling rather than encircling in pairs lesion.

Recent comparative trials of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) suggest that lower is better and that reducing low-density lipoprotein cholesterol (LDL-C) levels to below 100 mg/dL can provide additional clinical benefit. Non–high-density lipoprotein cholesterol (non–HDL-C) contains more atherogenic cholesterol than LDL-C and is considered a more accurate measurement of the total amount of atherogenic particles in the circulation. Therefore, the principle that “lower is better” may also apply to lowering levels of non–HDL-C. In persons with high triglycerides (200–499 mg/dL), LDL-C remains the primary target of therapy, but non–HDL-C is an important secondary therapeutic target. Non–HDL-C is strongly correlated with small dense LDL as well as apolipoprotein B, an established predictor of cardiovascular disease risk. Current evidence indicates that statins not only rapidly and dramatically reduce LDL-C, but also have a similar effect on non–HDL-C, and that the greater the reduction in LDL-C, the greater will be the reduction in non–HDL-C. (Prev Cardiol. 2005;8:173–177) © 2005 Le Jacq Ltd.

Dynamic Pulmonary Vein Analysis. Introduction: Anatomically guided left atrial ablation is used increasingly for treatment of atrial fibrillation (AF). Three-dimensional mapping systems used for pulmonary veins (PV) encircling ablation procedures anticipate a stable size and position of the PV orifice. The aim of the current study was therefore to analyze changes of PV orifice size and location throughout the cardiac cycle using cine magnetic resonance imaging (MRI). Methods and Results: Twenty-five healthy volunteers were studied using a 1.5 Tesla MRI system. MR angiograms were acquired with a breath-hold three-dimensional fast-spoiled gradient-echo imaging (3D FSPGR) sequence in the coronal plane before and after gadolinium injection. Maximum intensity projections and multiplanar reformations were performed to reconstruct images of the PV. Bright blood cine imaging in the axial view was acquired by a steady-state free precession pulse sequence. Twenty bright blood images were obtained per cardiac cycle. The axial (anterior-posterior) PV orifice diameter was measured in all 20 images. For analysis of PV movement the location of the orifice posterior edge was plotted on scale paper. PV orifice size depends on the stage of the cardiac cycle with the largest diameter in late atrial diastole and a mean decrease of 32.5% during atrial systole. Location changes of the PV orifice are in the range of up to 7.2 mm and larger in the coronal (lateral-medial) than in the sagittal (anterior-posterior) direction. Conclusion: PV orifice size and location is not as stable as anticipated by three-dimensional mapping systems used for PV encircling left atrial ablation procedures. RF application close to the presumed orifice location should therefore be avoided to minimize the risk of PV stenosis.

The Framingham risk estimation (FRE) serves as the basis for identifying which asymptomatic adults should be treated with aspirin and lipid-lowering therapy in primary prevention. However, the FRE generally yields low estimates of 10-year "hard" coronary heart disease (CHD) event risk with few women (70 years) qualifying for preventive pharmacologic therapy despite relatively high lifetime risk. We postulated that traditional risk factor assessment might fail to identify a sizeable portion of women with a sibling history for premature CHD as having advanced subclinical atherosclerosis.We studied 102 asymptomatic women (mean age 51 +/- 7 years) who were the sisters of a proband hospitalized with documented premature CHD. Participants underwent risk factor assessment and multidetector computed tomography for coronary artery calcium (CAC) scoring. Based on FRE prediction of 10-year risk for hard CHD events, participants were classified as low risk (10%) (n = 100), intermediate risk (10%-20%) (n = 2), or high risk (20%) (n = 0). Significant subclinical atherosclerosis was defined as age-sex adjusted75th percentile CAC scores.Ninety-eight percent were at low risk (mean FRE of only 2% +/- 2%). However, 40% had detectable CAC, 12% had CAC100, and 6% had CACor = 400. Based on CAC score percentiles, 32% had significant subclinical atherosclerosis and 17% ranked above the 90th percentile.Among women classified as low risk by FRE, a third had significant subclinical atherosclerosis. Sisters of probands with premature CHD appear to be a high-risk group and may warrant noninvasive screening for subclinical atherosclerosis to appropriately target individuals for more aggressive primary prevention therapy than what is currently recommended.

Cardiovascular disease (CVD) is the leading cause of death in the United States, resulting in increased awareness of the preventive importance of regular physical activity. Because athletes are considered physically fit, occurrence of sudden athlete death from CVD is perplexing. Regular intense physical activity can cause changes to the cardiovascular system that mimic known CVD processes. Therefore, screening of athletes for conditions that may increase risk for sudden cardiac death (SCD) is challenging. This article focuses on this problem, discussing the athlete's heart, SCD and associated CV conditions, and preparticipation screening. We also review recommendations of the 26th Bethesda Conference on determining eligibility for competition in athletes with known CV abnormalities, and how the recommendations relate to individual disease processes.

Introduction: The aims of this study were to identify predictors of recurrence after catheter ablation of atrial fibrillation (AF) and to report the safety and efficacy of catheter ablation of AF using an irrigated-tip ablation catheter. Methods and Results: Seventy-five consecutive patients (51 men [68%]; age 54 ± 13 years) with symptomatic drug-refractory paroxysmal (42 patients), persistent (21 patients), or permanent (12 patients) AF underwent catheter ablation of AF using an irrigated-tip ablation catheter and a standard ablation strategy, which involved electrical isolation of all pulmonary veins (PVs) and creation of a cavotricuspid linear lesion. At 10.5 ± 7.5 months of follow-up following a single (n = 75) or redo ablation procedure (n = 11), 39 (52%) of the 75 patients were free of AF, 10 were improved (13%), and 26 had experienced no benefit from the ablation procedure (35%). Seventy-six percent of patients with paroxysmal AF were free from recurrent AF. The most significant complications were two episodes of pericardial tamponade, mitral valve injury in one patient, two strokes, and complete but asymptomatic PV stenosis in one patient. Cox proportional hazards multivariate regression analysis identified the presence of persistent AF, permanent AF, and age >50 years prior to the ablation are the only independent predictors of AF recurrence after the first PV isolation procedure. Conclusion: Catheter ablation of AF using a strategy involving isolation of all PVs and creation of a linear lesion in the cavotricuspid isthmus using cooled radiofrequency energy is associated with moderate efficacy and an important risk for complications. The best results of this procedure are achieved in the subset of patients who are younger than 50 years and have only paroxysmal AF. (J Cardiovasc Electrophysiol, Vol. 15, pp. 692-697, June 2004)

Introduction: Catheter ablation of the pulmonary veins (PVs) for prevention of recurrent atrial fibrillation requires precise anatomic information. We describe the characteristics of a new anatomic variant of PV anatomy using magnetic resonance angiography. Methods and Results: A 1.5-T magnetic resonance imaging system with a body coil or a torso phased-array coil was used before and after gadolinium injection. Magnetic resonance angiograms were acquired with a breath-hold three-dimensional fast spoiled gradient-echo imaging sequence in the coronal plane. Three-dimensional reconstruction with maximum intensity projections and multiplanar reformations was performed. A newly described variant PV ascending from the roof of the left atrium was found in 3 of 91 subjects. The mean ostial diameter of the roof PV was 7 ± 2 mm, the mean distance from the ostium to the first branching point was 22 ± 8.5 mm, and the mean distance to the right superior PV was 3.3 ± 0.6 mm. Conclusion: We refer to the newly described variant of PV anatomy as the “right top pulmonary vein.” It is important to be aware of this anatomic pattern to avoid inadvertent catheter intubation, which can result in misleading mapping results and PV stenosis. (J Cardiovasc Electrophysiol, Vol. 15, pp. 538-543, May 2004)

Mucormycosis is rare, and its incidence is difficult to calculate accurately. The causative agents are saprophytic fungi of the class Zygomyces, order Mucorales. Rhinocerebral disease, the most common form, which accounts for more than half of the cases, is associated with diabetes and ketoacidosis. The syndromes of pulmonary and disseminated mucormycosis are usually seen in neutropenic patients. The development of primary cutaneous infection from mucormycosis depends on underlying systemic illness and local host factors. Although mucormycosis is infrequent, more cases of cutaneous infections are being noted, especially in the compromised host. We describe a case of Mucor as a predominant organism in a polymicrobial soft tissue infection in a diabetic patient with necrotizing fasciitis.

While much is known about general controls over axon guidance of broad classes of projection neurons (those with long-distance axonal connections), molecular controls over specific axon targeting by distinct neuron subtypes are poorly understood. Corticospinal motor neurons (CSMN) are prototypical and clinically important cerebral cortex projection neurons; they are the brain neurons that degenerate in amyotrophic lateral sclerosis (ALS) and related motor neuron diseases, and their injury is central to the loss of motor function in spinal cord injury. Primary culture of purified immature murine CSMN has been recently established, using either fluorescence-activated cell sorting (FACS) or immunopanning, enabling a previously unattainable level of subtype-specific investigation, but the resulting number of CSMN is quite limiting for standard approaches to study axon guidance. We developed a microfluidic system specifically designed to investigate axon targeting of limited numbers of purified CSMN and other projection neurons in culture. The system contains two chambers for culturing target tissue explants, allowing for biologically revealing axonal growth "choice" experiments. This device will be uniquely enabling for investigation of controls over axon growth and neuronal survival of many types of neurons, particularly those available only in limited numbers.