An Unmanned Surface Vehicle (USV): Development of an Autonomous Boat with a Sensor Integration System for Bathymetric Surveys.

A reliable yet economical unmanned surface vehicle (USV) has been developed for the bathymetric surveying of lakes. The system combines an autonomous navigation framework, environmental sensors, and a multibeam echosounder to collect submerged topography, temperature, and wind speed and monitor the vehicle's status during prescribed path-planning missions. The main objective of this research is to provide a methodological framework to build an autonomous boat with independent decision-making, efficient control, and long-range navigation capabilities. Integration of sensors with navigation control enabled the automatization of position, orientation, and velocity. A solar power integration was also tested to control the duration of the autonomous missions. The results of the solar power compared favorably with those of the standard LiPO battery system. Extended and autonomous missions were achieved with the developed platform, which can also evaluate the danger level, weather circumstances, and energy consumption through real-time data analysis. With all the incorporated sensors and controls, this USV can make self-governing decisions and improve its safety. A technical evaluation of the proposed vehicle was conducted as a measurable metric of the reliability and robustness of the prototype. Overall, a reliable, economic, and self-powered autonomous system has been designed and built to retrieve bathymetric surveys as a first step to developing intelligent reconnaissance systems that combine field robotics with machine learning to make decisions and adapt to unknown environments.

Exome sequencing identifies rare LDLR and APOA5 alleles conferring risk for myocardial infarction.

Myocardial infarction (MI), a leading cause of death around the world, displays a complex pattern of inheritance. When MI occurs early in life, genetic inheritance is a major component to risk. Previously, rare mutations in low-density lipoprotein (LDL) genes have been shown to contribute to MI risk in individual families, whereas common variants at more than 45 loci have been associated with MI risk in the population. Here we evaluate how rare mutations contribute to early-onset MI risk in the population. We sequenced the protein-coding regions of 9,793 genomes from patients with MI at an early age (≤50 years in males and ≤60 years in females) along with MI-free controls. We identified two genes in which rare coding-sequence mutations were more frequent in MI cases versus controls at exome-wide significance. At low-density lipoprotein receptor (LDLR), carriers of rare non-synonymous mutations were at 4.2-fold increased risk for MI; carriers of null alleles at LDLR were at even higher risk (13-fold difference). Approximately 2% of early MI cases harbour a rare, damaging mutation in LDLR; this estimate is similar to one made more than 40 years ago using an analysis of total cholesterol. Among controls, about 1 in 217 carried an LDLR coding-sequence mutation and had plasma LDL cholesterol > 190 mg dl(-1). At apolipoprotein A-V (APOA5), carriers of rare non-synonymous mutations were at 2.2-fold increased risk for MI. When compared with non-carriers, LDLR mutation carriers had higher plasma LDL cholesterol, whereas APOA5 mutation carriers had higher plasma triglycerides. Recent evidence has connected MI risk with coding-sequence mutations at two genes functionally related to APOA5, namely lipoprotein lipase and apolipoprotein C-III (refs 18, 19). Combined, these observations suggest that, as well as LDL cholesterol, disordered metabolism of triglyceride-rich lipoproteins contributes to MI risk.

A Journey through Genetic Architecture and Predisposition of Coronary Artery Disease.

INTRODUCTION: To halt the spread of coronary artery disease (CAD), the number one killer in the world, requires primary prevention. Fifty percent of all Americans are expected to experience a cardiac event; the challenge is identifying those at risk. 40 to 60% of predisposition to CAD is genetic. The first genetic risk variant, 9p21, was discovered in 2007. Genome-Wide Association Studies has since discovered hundreds of genetic risk variants. The genetic burden for CAD can be expressed as a single number, Genetic Risk Score (GRS). Assessment of GRS to risk stratify for CAD was superior to conventional risk factors in several large clinical trials assessing statin therapy, and more recently in a population of nearly 500,000 (UK Biobank). Studies were performed based on prospective genetic risk stratification for CAD. These studies showed that a favorable lifestyle was associated with a 46% reduction in cardiac events and programmed exercise, a 50% reduction in cardiac events. Genetic risk score is superior to conventional risk factors, and is markedly attenuated by lifestyle changes and drug therapy. Genetic risk can be determined at birth or any time thereafter. CONCLUSION: Utilizing the GRS to risk stratify young, asymptomatic individuals could provide a paradigm shift in the primary prevention of CAD and significantly halt its spread.

Effect of high-pressure-jet processing on the physiochemical properties of low-fat ice cream mix.

The objective of this study was to use high-pressure-jet (HPJ) processing to produce functional properties in a low-fat (4.5% fat) ice cream mix similar to those seen when emulsifiers are used. Ice cream mix or serum (nonfat portion of the ice cream mix) were subjected to 200 or 400 MPa HPJ processing and compared with a non-HPJ-treated control. A similar non-HPJ-treated formulation but containing polysorbate 80 (0.075% wt/wt) was also used as a control. The mix samples were characterized in terms of their particle size, density, flow properties, stability, crystallization kinetics, and fat-protein interactions. The sample from the mix subjected to 400 MPa HPJ processing (HPJ-M-400) had increased consistency coefficient (5°C; 228 ± 102.7 mPa·s) and particle size (D[4,3]; 16.0 ± 2.5 µm) compared with the non-HPJ-treated control sample, with viscosity and particle size (volume-moment mean diameter, D[4,3]) values of 7.5 ± 0.4 mPa·s and 0.50 ± 0.1 µm, respectively. These differences were attributed to an increase in casein-fat interactions and casein-casein interactions caused by the 400 MPa HPJ treatment, which were observed using confocal scanning laser microscopy and inferred from an increase in protein and fat concentrations in the sediment after ultracentrifugation. Interestingly, the density of HPJ-M-400 was also lower (0.79 ± 0.17 g/mL) than that of the control (1.04 ± 0.00 g/mL) because bubbles were trapped within these complexes. The large casein-fat complexes formed in the HPJ-M-400 sample also appeared to act as steric barriers that slowed ice crystal growth during quiescent freezing. The alterations in physiochemical properties and apparent ice crystal growth induced by the 400 MPa treatment of low-fat ice cream mix have many potential applications, including clean-label confections.

Enhanced Controllability of Fries Rearrangements Using High-Resolution 3D-Printed Metal Microreactor with Circular Channel.

High-resolution 3D-printed stainless steel metal microreactors (3D-PMRs) with different cross-sectional geometry are fabricated to control ultrafast intramolecular rearrangement reactions in a comparative manner. The 3D-PMR with circular channel demonstrates the improved controllability in rapid Fries-type rearrangement reactions, because of the superior mixing efficiency to rectangular cross-section channels (250 µm × 125 µm) which is confirmed based on the computational flow dynamics simulation. Even in case of very rapid intramolecular rearrangement of sterically small acetyl group occurring in 333 µs of reaction time, the desired intermolecular reaction can outpace to the undesired intramolecular rearrangement using 3D-PMR to result in high conversion and yield.

Runs of Homozygosity: Association with Coronary Artery Disease and Gene Expression in Monocytes and Macrophages.

Runs of homozygosity (ROHs) are recognized signature of recessive inheritance. Contributions of ROHs to the genetic architecture of coronary artery disease and regulation of gene expression in cells relevant to atherosclerosis are not known. Our combined analysis of 24,320 individuals from 11 populations of white European ethnicity showed an association between coronary artery disease and both the count and the size of ROHs. Individuals with coronary artery disease had approximately 0.63 (95% CI: 0.4-0.8) excess of ROHs when compared to coronary-artery-disease-free control subjects (p = 1.49 × 10(-9)). The average total length of ROHs was approximately 1,046.92 (95% CI: 634.4-1,459.5) kb greater in individuals with coronary artery disease than control subjects (p = 6.61 × 10(-7)). None of the identified individual ROHs was associated with coronary artery disease after correction for multiple testing. However, in aggregate burden analysis, ROHs favoring increased risk of coronary artery disease were much more common than those showing the opposite direction of association with coronary artery disease (p = 2.69 × 10(-33)). Individual ROHs showed significant associations with monocyte and macrophage expression of genes in their close proximity-subjects with several individual ROHs showed significant differences in the expression of 44 mRNAs in monocytes and 17 mRNAs in macrophages when compared to subjects without those ROHs. This study provides evidence for an excess of homozygosity in coronary artery disease in outbred populations and suggest the potential biological relevance of ROHs in cells of importance to the pathogenesis of atherosclerosis.

Genetically determined height and coronary artery disease.

BACKGROUND: The nature and underlying mechanisms of an inverse association between adult height and the risk of coronary artery disease (CAD) are unclear. METHODS: We used a genetic approach to investigate the association between height and CAD, using 180 height-associated genetic variants. We tested the association between a change in genetically determined height of 1 SD (6.5 cm) with the risk of CAD in 65,066 cases and 128,383 controls. Using individual-level genotype data from 18,249 persons, we also examined the risk of CAD associated with the presence of various numbers of height-associated alleles. To identify putative mechanisms, we analyzed whether genetically determined height was associated with known cardiovascular risk factors and performed a pathway analysis of the height-associated genes. RESULTS: We observed a relative increase of 13.5% (95% confidence interval [CI], 5.4 to 22.1; P<0.001) in the risk of CAD per 1-SD decrease in genetically determined height. There was a graded relationship between the presence of an increased number of height-raising variants and a reduced risk of CAD (odds ratio for height quartile 4 versus quartile 1, 0.74; 95% CI, 0.68 to 0.84; P<0.001). Of the 12 risk factors that we studied, we observed significant associations only with levels of low-density lipoprotein cholesterol and triglycerides (accounting for approximately 30% of the association). We identified several overlapping pathways involving genes associated with both development and atherosclerosis. CONCLUSIONS: There is a primary association between a genetically determined shorter height and an increased risk of CAD, a link that is partly explained by the association between shorter height and an adverse lipid profile. Shared biologic processes that determine achieved height and the development of atherosclerosis may explain some of the association. (Funded by the British Heart Foundation and others.).

Genetics in the prevention and management of coronary artery disease.

PURPOSE OF REVIEW: The current review is to describe the genetic risk variants that have been discovered predisposing to coronary artery disease (CAD) and how they are utilized to stratify for risk of CAD. RECENT FINDINGS: Over 90 genetic risk variants have been discovered that predispose to risk for CAD. SUMMARY: The total genetic risk burden for CAD is proportional to the number of risk variants inherited and can be combined into a single number referred to as the genetic risk score (GRS). GRS has been utilized in multiple studies and shown to be more effective in risk stratification for CAD than conventional risk factors. There is a major advantage to risk stratification based on the GRS since the risk can be determined at birth or anytime throughout one's lifetime since the individual's DNA does not change. Widespread application of the GRS is likely to enable a paradigm shift in the primary prevention of CAD.

Genetic stratification for primary prevention of coronary artery disease.

PURPOSE OF REVIEW: To review the literature showing genetic risk variants is a reliable means of stratifying for risk of CAD for primary prevention. RECENT FINDINGS: Over 90 genetic risk variants have been discovered that predispose to CAD. Results of several studies show that these risk variants effectively stratify for risk of CAD in asymptomatic individuals. SUMMARY: The total individual genetic risk can be summarized into a single number referred to as the genetic risk score (GRS). The GRS unlike the Framingham Risk Score is not dependent on age and independent of conventional risk factors. As DNA does not change during one's lifetime the GRS can be calculated at birth or any time thereafter. Furthermore, the GRS has been shown to provide superior discriminatory power in selecting individuals who will benefit most from lifestyle changes or statin therapy. A prospective study showed individuals with high GRS and a favorable lifestyle was associated with significant reduction of cardiac events compared with an unfavorable lifestyle. Furthermore, the study shows inherited risk can be reduced analogous to reduction of risk form acquired and environmental factors. The use of GRS to stratify for risk of CAD in asymptomatic individuals could transform primary prevention worldwide.

The Identification of a Threshold of Long Work Hours for Predicting Elevated Risks of Adverse Health Outcomes.

Working long hours has been associated with adverse health outcomes. However, a definition of long work hours relative to adverse health risk has not been established. Repeated measures of work hours among approximately 2,000 participants from the Panel Study of Income Dynamics (1986-2011), conducted in the United States, were retrospectively analyzed to derive statistically optimized cutpoints of long work hours that best predicted three health outcomes. Work-hours cutpoints were assessed for model fit, calibration, and discrimination separately for the outcomes of poor self-reported general health, incident cardiovascular disease, and incident cancer. For each outcome, the work-hours threshold that best predicted increased risk was 52 hours per week or more for a minimum of 10 years. Workers exposed at this level had a higher risk of poor self-reported general health (relative risk (RR) = 1.28; 95% confidence interval (CI): 1.06, 1.53), cardiovascular disease (RR = 1.42; 95% CI: 1.24, 1.63), and cancer (RR = 1.62; 95% CI: 1.22, 2.17) compared with those working 35-51 hours per week for the same duration. This study provides the first health risk-based definition of long work hours. Further examination of the predictive power of this cutpoint on other health outcomes and in other study populations is needed.

Consumption of Bifidobacterium animalis subsp. lactis BB-12 in yogurt reduced expression of TLR-2 on peripheral blood-derived monocytes and pro-inflammatory cytokine secretion in young adults.

PURPOSE: Probiotic bacteria modulate immune parameters and inflammatory outcomes. Emerging evidence demonstrates that the matrix used to deliver probiotics may influence the efficacy of probiotic interventions in vivo. The aims of the current study were to evaluate (1) the effect of one species, Bifidobacterium animalis subsp. lactis BB-12 at a dose of log10 ± 0.5 CFUs/day on immune responses in a randomized, partially blinded, 4-period crossover, free-living study, and (2) whether the immune response to BB-12 differed depending on the delivery matrix. METHODS: Healthy adults (n = 30) aged 18-40 years were recruited and received four treatments in a random order: (A) yogurt smoothie alone; smoothie with BB-12 added (B) before or (C) after yogurt fermentation, or (D) BB-12 given in capsule form. At baseline and after each 4-week treatment, peripheral blood mononuclear cells (PBMCs) were isolated, and functional and phenotypic marker expression was assessed. RESULTS: BB-12 interacted with peripheral myeloid cells via Toll-like receptor 2 (TLR-2). The percentage of CD14+HLA-DR+ cells in peripheral blood was increased in male participants by all yogurt-containing treatments compared to baseline (p = 0.0356). Participants who consumed yogurt smoothie with BB-12 added post-fermentation had significantly lower expression of TLR-2 on CD14+HLA-DR+ cells (p = 0.0186) and reduction in TNF-α secretion from BB-12- (p = 0.0490) or LPS-stimulated (p = 0.0387) PBMCs compared to baseline. CONCLUSIONS: These findings not only demonstrate a potential anti-inflammatory effect of BB-12 in healthy adults, but also indicate that the delivery matrix influences the immunomodulatory properties of BB-12.

Safety of Bifidobacterium animalis Subsp. Lactis (B. lactis) Strain BB-12-Supplemented Yogurt in Healthy Children.

OBJECTIVES: Probiotics are live microorganisms that may provide health benefits to the individual when consumed in sufficient quantities. For studies conducted on health or disease endpoints on probiotics in the United States, the Food and Administration has required those studies to be conducted as investigational new drugs. This phase I, double-blinded, randomized, controlled safety study represents the first requirement of this pathway. The purpose of the study was to determine the safety of Bifidobacterium animalis subsp. lactis (B lactis) strain BB-12 (BB-12)-supplemented yogurt when consumed by a generally healthy group of children. The secondary aim was to assess the effect of BB-12-supplemented yogurt on the gut microbiota of the children. METHODS: Sixty children ages 1 to 5 years were randomly assigned to consume 4 ounces of either BB-12-supplemented yogurt or nonsupplemented control yogurt daily for 10 days. The primary outcome was to assess safety and tolerability, as determined by the number of reported adverse events. RESULTS: A total of 186 nonserious adverse events were reported, with no significant differences between the control and BB-12 groups. No significant changes due to probiotic treatment were observed in the gut microbiota of the study cohort. CONCLUSIONS: BB-12-supplemented yogurt is safe and well-tolerated when consumed by healthy children. The present study will form the basis for future randomized clinical trials investigating the potential effects of BB-12-supplemented yogurt in different disease states.

Effects of Bifidobacterium animalis subsp. lactis BB-12® on the lipid/lipoprotein profile and short chain fatty acids in healthy young adults: a randomized controlled trial.

BACKGROUND: Some probiotics have hypocholesterolemic effects in animal studies, which are mediated, in part, by increases in fecal short chain fatty acids (SCFAs). Clinical trials of probiotics on lipids/lipoproteins are inconsistent. OBJECTIVE: We examined the effects of Bifidobacterium animalis subsp. lactis BB-12® (BB-12®) (3.16 × 109 CFUs/day) on lipids and lipoproteins and fecal excretion of SCFAs in healthy adults. METHODS: In a randomized, partially blinded, 4-period, crossover study, 30 adults (11 men, 19 women) aged 18-40 years were randomly assigned to: 1) yogurt smoothie with no BB-12® (YS), 2) yogurt smoothie with BB-12® added pre-fermentation (PRE), 3) yogurt smoothie with BB-12® added post-fermentation (POST), 4) BB-12® containing capsule (CAP). We measured serum lipids/lipoproteins, glucose, insulin, C-reactive protein (CRP), and fecal SCFAs at baseline and after each treatment period. RESULTS: Total cholesterol (TC), LDL cholesterol (LDL-C), HDL cholesterol (HDL-C), and triglycerides (TGs) did not differ after the PRE, POST, and CAP periods versus the YS or between treatments. Compared to baseline, fecal acetate was significantly increased after the YS (Δ = 211.89 ± 75.87 µg/g, P = 0.007) and PRE (Δ = 204.98 ± 75.70 µg/g, P = 0.009) periods. The percent increase in fecal acetate was significantly greater after the YS versus the POST period (52.2 ± 13.2% vs. 24.5 ± 13.2%, P = 0.023). Fecal total SCFAs, propionate and butyrate did not differ between treatment periods. Fecal total SCFAs were negatively associated with TC (r = -0.22, P = 0.01), LDL-C (r = -0.24, P = 0.004), age (r = -0.33, P < 0.001), and waist circumference (r = -0.25, P = 0.003). CONCLUSIONS: BB-12® supplementation did not improve lipids, lipoproteins and total and individual fecal SCFAs. Fecal SCFAs were negatively associated with TC, LDL-C, age, and waist circumference. TRIAL REGISTRATION: This trial was registered at clinicaltrials.gov as NCT01399996 .

Effect of fat content on the physical properties and consumer acceptability of vanilla ice cream.

Ice cream is a complex food matrix that contains multiple physical phases. Removal of 1 ingredient may affect not only its physical properties but also multiple sensory characteristics that may or may not be important to consumers. Fat not only contributes to texture, mouth feel, and flavor, but also serves as a structural element. We evaluated the effect of replacing fat with maltodextrin (MD) on select physical properties of ice cream and on consumer acceptability. Vanilla ice creams were formulated to contain 6, 8, 10, 12, and 14% fat, and the difference was made up with 8, 6, 4, 2, and 0% maltodextrin, respectively, to balance the mix. Physical characterization included measurements of overrun, apparent viscosity, fat particle size, fat destabilization, hardness, and melting rate. A series of sensory tests were conducted to measure liking and the intensity of various attributes. Tests were also conducted after 19 weeks of storage at -18°C to assess changes in acceptance due to prolonged storage at unfavorable temperatures. Then, discrimination tests were performed to determine which differences in fat content were detectable by consumers. Mix viscosity decreased with increasing fat content and decreasing maltodextrin content. Fat particle size and fat destabilization significantly increased with increasing fat content. However, acceptability did not differ significantly across the samples for fresh or stored ice cream. Following storage, ice creams with 6, 12, and 14% fat did not differ in acceptability compared with fresh ice cream. However, the 8% fat, 6% MD and 10% fat, 4% MD ice creams showed a significant drop in acceptance after storage relative to fresh ice cream at the same fat content. Consumers were unable to detect a difference of 2 percentage points in fat level between 6 and 12% fat. They were able to detect a difference of 4 percentage points for ice creams with 6% versus 10%, but not for those with 8% versus 12% fat. Removing fat and replacing it with maltodextrin caused minimal changes in physical properties in ice cream and mix and did not change consumer acceptability for either fresh or stored ice cream.

The Stability of Retrospective Child Sexual Abuse Reports and its Association With Problem Avoidance.

Due to reliability problems, use of retrospective measurement to assess child sexual abuse has long concerned researchers. Possible psychosocial causes of these reliability issues-including problem avoidance-have not been thoroughly studied. We tested the reliability of retrospective child sexual abuse measurement in a nationally representative sample of 12,438 adults over two periods (2001-2002, 2007-2008), assessed sex differences in reliability, and examined whether reliability depends on problem avoidance tendencies. Nearly three-fourths of child sexual abuse cases in the former wave were not again reported, and two-thirds of child sexual abuse cases in the latter wave were not previously reported. Females were more likely to report CSA later if reported previously (OR = 5.11). Participants who reported child sexual abuse in the former wave but not the latter were more avoidant than consistent reporters (3.13 versus 2.77). Our findings suggest that females may report child sexual abuse more consistently. Furthermore, inconsistent reporting may indicate problem avoidance. Suggestions for researchers and practitioners are discussed.

High Sensitivity, Wearable, Piezoresistive Pressure Sensors Based on Irregular Microhump Structures and Its Applications in Body Motion Sensing.

A pressure sensor based on irregular microhump patterns has been proposed and developed. The devices show high sensitivity and broad operating pressure regime while comparing with regular micropattern devices. Finite element analysis (FEA) is utilized to confirm the sensing mechanism and predict the performance of the pressure sensor based on the microhump structures. Silicon carbide sandpaper is employed as the mold to develop polydimethylsiloxane (PDMS) microhump patterns with various sizes. The active layer of the piezoresistive pressure sensor is developed by spin coating PEDOT: PSS on top of the patterned PDMS. The devices show an averaged sensitivity as high as 851 kPa(-1) , broad operating pressure range (20 kPa), low operating power (100 nW), and fast response speed (6.7 kHz). Owing to their flexible properties, the devices are applied to human body motion sensing and radial artery pulse. These flexible high sensitivity devices show great potential in the next generation of smart sensors for robotics, real-time health monitoring, and biomedical applications.

Genetics-Current and Future Role in the Prevention and Management of Coronary Artery Disease.

PURPOSE OF REVIEW: The purpose of this study is to review genetic risk variants for coronary artery disease (CAD) and how they will change the management and prevention of CAD currently and in the future. RECENT FINDINGS: Through the efforts of international consortia, 58 genetic risk variants for CAD of genome-wide significance have been replicated in appropriate independent populations. Only one third of these variants mediate their risk through known conventional risk factors for CAD. Thus, unknown mechanisms contribute to CAD. Secondly, the genetic risk is proportional to the total number of risk variants rather than the intensity of any risk factor. Thirdly, the availability of the genetic risk variants enables one to perform Mendelian randomization (MR) studies since they are randomized at conception, not confounded, fixed for life, and can be used to determine if a risk factor is causative or just a marker. MR can also be used to determine the safety and efficacy of a gene product targeted for drug therapy. Genetic risk variants have been shown to successfully risk stratify for CAD in both primary and secondary preventions. Contrary to dogma, MR documents that plasma HDL-C is not protective of CAD. The use of genetic risk score (GRS) for CAD is shown to be more effective in risk stratifying for CAD than the Framingham risk score and independent of the conventional risk factors including family history. Furthermore, the GRS predicts the response to statin therapy in primary and secondary preventions. The use of GRS could represent a paradigm shift in the prevention of CAD.

Genetics of coronary artery disease.

There is almost no data on the genetics of acute coronary syndromes, so this review discusses primarily the 50 genetic risk variants associated with coronary artery disease that are of genome-wide significance in the discovery population and replicated in an independent population. All of these risk variants are extremely common with more than half occurring in >50% of the general population. They increased only minimally the relative risk for coronary artery disease. The most striking finding is that 35 of the 50 risk variants act independently of known risk factors, indicating there are several pathways yet to be appreciated, contributing to the pathogenesis of coronary atherosclerosis and myocardial infarction. All of the genetic variants seem to act through atherosclerosis, except for the ABO blood groups, which show that A and B are associated with increased risk for myocardial infarction, mediated by a prolonged von Willebrand plasma half life leading to thrombosis. The potential molecular mechanisms of 9p21 are discussed, including cell cycle kinase inhibitors. Discovery of risk variants associated with PCSK9 has led to the development of novel treatment for plasma low-density lipoprotein cholesterol. A monoclonal antibody inhibiting PCSK9 has already undergone phase I and II clinical trials, showing it is a potent inhibitor of low-density lipoprotein cholesterol and is mediated through more rapid removal of low-density lipoprotein cholesterol from the plasma. This therapy complements that of statin therapy, which inhibits the synthesis of cholesterol. The benefits of Mendelian randomization to assess safety and efficacy and their limitations are discussed along with future directions.