Effects of epicatechin on cardiovascular function in middle-aged diet-induced obese rat models of metabolic syndrome.

The current study aimed to investigate the cardiovascular effects of epicatechin, a flavonoid found in green tea and cocoa, in attenuating complications associated with metabolic syndrome in diet-induced obese rats. Male Wistar-Kyoto (WKY) rats aged 16 weeks were fed either standard rat chow or given a high-fat-high-carbohydrate (HFHC) diet for 20 weeks. Epicatechin treatment (5 mg/kg/d) was administered to a subset of WKY rats commencing at week 8 of the 20 week HFHC feeding period. Body weights, food, water and energy intakes, blood pressure, heart rate and glucose tolerance were measured throughout the treatment period. Oxidative stress and inflammatory markers, lipid levels, cardiac collagen deposition, cardiac electrical function, aortic and mesenteric vessel reactivity were examined after the treatment. Twenty weeks of HFHC feeding in WKY rats resulted in the development of metabolic syndrome indicated by the presence of abdominal obesity, dyslipidaemia, glucose intolerance and increased blood pressure. Epicatechin treatment was found to enhance the oxidative stress status in HFHC groups through an increase in serum nitric oxide levels and a decrease in 8-isoprostane concentrations. Furthermore, WKY-HFHC rats displayed a decrease in IL-6 levels. The lipid profiles in HFHC groups showed improvement, with a decrease in LDL-cholesterol and TAG and an increase in HDL-cholesterol levels observed in WKY-HFHC rats. However, epicatechin was not effective in preventing weight gain, glucose intolerance or hypertension in HFHC fed rats. Overall, the results of this study suggest that epicatechin has the potential to improve the underlying mechanisms associated with metabolic syndrome in obese rats.

The impact of the Community Resiliency Model (CRM) on the mental well-being of youth at risk for violence: A study protocol.

PURPOSE: Assess the effect of a brief, somatic awareness resiliency training, the Community Resiliency Model (CRM)®, on the mental well-being of incarcerated youth. SPECIFIC AIM 1: Explore the relationship between youth demographics and justice history to baseline well-being scores and response to treatment. Q1: How do personal variables, including length of time in juvenile justice, relate to baseline mental health scores and response to the wellness skills intervention? SPECIFIC AIM 2: Examine the mental health scores over time to determine effectiveness of participating in CRM training while incarcerated. Hypothesis 1: Well-being scores will increase after CRM training. Hypothesis 2: Emotional distress will decrease after participating in a CRM training. Hypothesis 3: Pro-social feelings and behaviors will increase after participating in a CRM training.

Perindopril prevents development of obesity and hypertension in middle aged diet-induced obese rat models of metabolic syndrome.

AIMS: The therapeutic properties of anti-hypertensive medications that extend beyond blood pressure lowering have started to become important clinical targets in recent years. This study aimed to assess the cardioprotective effects of perindopril in attenuating complications associated with metabolic syndrome in diet induced obese rats. MAIN METHODS: Male Wistar-Kyoto (WKY) rats aged 16 weeks were fed either standard rat chow (SC) or given a high-fat-high-carbohydrate (HFHC) diet for 20 weeks. Perindopril treatment (1 mg/kg/day) was administered to a subset of WKY rats commencing at week 8 of the 20 week HFHC feeding period. Body weights, food, water and energy intakes, blood pressure, heart rate and glucose tolerance were measured throughout the treatment period. Oxidative stress and inflammatory markers, lipid levels, cardiac collagen deposition, vascular function, aortic and cardiac electrical function were examined after the treatment. KEY FINDINGS: WKY rats developed metabolic syndrome after 20 weeks of HFHC feeding, evidenced by the presence of abdominal obesity, dyslipidaemia, glucose intolerance and hypertension. Perindopril treatment prevented the development of obesity and hypertension in WKY-HFHC. Perindopril improved blood lipid profiles in HFHC rats with decreases in LDL cholesterol, triglycerides and total cholesterol. Type I collagen levels were decreased in WKY-HFHC rats along with decreases in left ventricle mass. Perindopril treated rats also showed improved cardiac electrical function indicated by decreases in action potential at 90 % of repolarisation in WKY-HFHC rats. SIGNIFICANCE: These results show that perindopril has a profound effect on preventing the development of metabolic syndrome in animals fed a HFHC diet.

Bactericidal effects of high-energy visible light on common otitis media pathogens.

AIMS: This study assessed the use of high-energy, visible light on the survival rates of three bacteria commonly found in middle ear infections (i.e. otitis media; Streptococcus pneumoniae, Moraxella catarrhalis and Haemophilus influenzae). METHOD AND RESULTS: Bacteria were cultured and then subjected to a single, 4-h treatment of 405 nm wavelength light at two different intensities. All three bacteria species were susceptible to the light at clinically significant rates (>99.9% reduction). Bacteria were susceptible to the high-energy visible (HEV) light in a dose-dependent manner (lower survival rates with increased intensity and duration of exposure). CONCLUSIONS: The results suggest that HEV light may provide a non-surgical, non-pharmaceutical approach to the therapeutic treatment of otitis media. SIGNIFICANCE AN IMPACT OF THE STUDY: Given the growing concerns surrounding antibiotic resistance, this study demonstrates a rapid, alternative method for effective inactivation of bacterial pathogens partly responsible for instances of otitis media.

A Pilot Study to Adapt a Trauma-Informed, Mindfulness-Based Yoga Intervention for Justice-Involved Youth.

The purpose of the present study was to adapt and pilot a trauma-informed, mindfulness-based yoga (TIMBY) intervention focused on enhancing self-regulation among youth in the Georgia Department of Juvenile Justice system. In this article we (1) describe the process by which we systematically adapted an evidence-based protocol specifically for this population, (2) describe the nature of and rationale for those adaptations, (3) present some preliminary qualitative findings based on interviews with youth participants, and (4) briefly summarize how the adapted protocol will be evaluated in the subsequent feasibility trial. The iterative drafting and revision process involved modifications to a well-established, protocolized Trauma-Informed Yoga program and was identified by the project advisory board and t h rough formal interviews with intervention staff. Qualitative interviews were conducted with youth participants concerning intervention impact, credibility, and satisfaction. Several needed modifications were identified so that the intervention would be contextually appropriate for justice-involved youth. Thirty youth were enrolled in the pilot study: 77% we re Non - Hispanic Black/African-American, 18% were Non-Hispanic White, and 5% were Hispanic White. The average age was 16.45 years (range 14-20). The youth consistently reported satisfaction with the sessions and positive beliefs about how the sessions were helping them with a range of physical and psychological/ emotional challenges. Adaptations to the protocol in the present study highlight how mindfulness-based interventions for justice-involved youth need to consider what is both developmentally suitable for youth and appropriate in a justice setting. A feasibility study using this revised TIMBY protocol is underway at four Georgia Department of Juvenile Justice facilities to formally identify the barriers and facilitators to implementation for the present study and a future, larger-scale trial.

Late Pleistocene to early Holocene high-quality quartz crystal procurement from the Valiente quarry workshop site (32°S, Chile, South America).

The procurement of high-quality lithic resources is amongst the most indicative processes of decision-making in the archaeology of early human groups peopling the Americas. Directly dated deposits from quarry workshops have been absent of the late Pleistocene record of South America. We present the results of the excavations of a high-quality translucent quartz crystal workshop that yielded radiocarbon-dated coherently layered stratigraphic deposits that shed light into the behavior of the initial stages of lithic procurement. Based on a detailed analysis of the context of the Valiente site (32° S, Chile, South America), we discuss the stages of bifacial production of point technology. The deposit produced evidence of cumulative occupations over the period between 12,630 and 11,320 calibrated years before present. This ~1,300-year span is coincidental with a major environmental step-wise drying trend as indicated by the local and regional pollen records. Furthermore, it is synchronous to the process in which natural landscapes became the earliest taskscapes in the region, thereby encompassing major cultural changes related to the organization of the land use. These results are discussed in the frame of contemporaneous archaeological data to discuss specific aspects of technology and decision-making of the earliest settlers of South America.

(-)-Epicatechin Reduces Blood Pressure and Improves Left Ventricular Function and Compliance in Deoxycorticosterone Acetate-Salt Hypertensive Rats.

(−)-Epicatechin (E) is a flavanol found in green tea and cocoa and has been shown to attenuate tumour necrosis factor alpha (TNF-α)-mediated inflammation, improve nitric oxide levels, promote endothelial nitric oxide synthase (eNOS) activation and inhibit NADPH oxidase. This study investigated the effect of 28 days of low epicatechin dosing (1 mg/kg/day) on the cardiovascular function of deoxycorticosterone acetate (DOCA)-salt hypertensive rats. Wistar rats (n = 120, 8 weeks of age) underwent uninephrectomy and were randomised into four groups (uninephrectomy (UNX), UNX + E, DOCA, DOCA + E). DOCA and DOCA + E rats received 1% NaCl drinking water along with subcutaneous injections of 25 mg deoxycorticosterone-acetate (in 0.4 mL of dimethylformamide) every fourth day. UNX + E and DOCA + E rats received 1 mg/kg/day of epicatechin by oral gavage. Single-cell micro-electrode electrophysiology, Langendorff isolated-heart assessment and isolated aorta and mesenteric organ baths were used to assess cardiovascular parameters. Serum malondialdehyde concentration was used as a marker of oxidative stress. Myocardial stiffness was increased and left ventricular compliance significantly diminished in the DOCA control group, and these changes were attenuated by epicatechin treatment (p < 0.05). Additionally, the DOCA + E rats showed significantly reduced blood pressure and malondialdehyde concentrations; however, there was no improvement in left ventricular hypertrophy, electrophysiology or vascular function. This study demonstrates the ability of epicatechin to reduce blood pressure, prevent myocardial stiffening and preserve cardiac compliance in hypertrophied DOCA-salt rat hearts.

Articular Cartilage: Injury Pathways and Treatment Options.

Articular cartilage injury and degeneration is a frequent occurrence in synovial joints. Treatment of these articular cartilage lesions are a challenge because this tissue is incapable of quality repair and/or regeneration to its native state. Nonoperative treatments endeavor to control symptoms, and include anti-inflammatory medication, viscosupplementation, bracing, orthotics, and activity modification. Techniques to stimulate the intrinsic repair (fibrocartilage) process include drilling, abrasion, and microfracture of the subchondral bone. Currently, the clinical biologic approaches to treat cartilage defects include autologous chondrocyte implantation, periosteal transfer, and osteochondral autograft or allograft transplantation. Newer strategies employing tissue engineering being studied involve the use of combinations of progenitor cells, bioactive factors, and matrices, and the use of focal synthetic devices. Many new and innovative treatments are being explored in this exciting field. However, there is a paucity of prospective, randomized controlled clinical trials that have compared the various techniques, treatment options, indications and efficacy.

Δ9-Tetrahydrocannabinol Prevents Cardiovascular Dysfunction in STZ-Diabetic Wistar-Kyoto Rats.

The aim of this study was to determine if chronic, low-dose administration of a nonspecific cannabinoid receptor agonist could provide cardioprotective effects in a model of type I diabetes mellitus. Diabetes was induced in eight-week-old male Wistar-Kyoto rats via a single intravenous dose of streptozotocin (65 mg kg-1). Following the induction of diabetes, Δ9-tetrahydrocannabinol was administered via intraperitoneal injection (0.15 mg kg-1 day-1) for an eight-week period until the animals reached sixteen weeks of age. Upon completion of the treatment regime, assessments of vascular reactivity and left ventricular function and electrophysiology were made, as were serum markers of oxidative stress and lipid peroxidation. Δ9-Tetrahydrocannabinol administration to diabetic animals significantly reduced blood glucose concentrations and attenuated pathological changes in serum markers of oxidative stress and lipid peroxidation. Positive changes to biochemical indices in diabetic animals conferred improvements in myocardial and vascular function. This study demonstrates that chronic, low-dose administration of Δ9-tetrahydrocannabinol can elicit antihyperglycaemic and antioxidant effects in diabetic animals, leading to improvements in end organ function of the cardiovascular system. Implications from this study suggest that cannabinoid receptors may be a potential new target for the treatment of diabetes-induced cardiovascular disease.

Alpha-adrenoceptor antagonism by Crassostrea gigas oyster extract inhibits noradrenaline-induced vascular contraction in Wistar rats.

OBJECTIVE: Crassostrea gigas oyster extract has been reported to have antioxidant, antihypertensive and lipid-lowering properties that may be useful for treating cardiovascular diseases. This study aimed to evaluate the effect of C. gigas oyster extract on cardiovascular function in tissues from healthy rats. METHODS: Single-cell microelectrode and isolated thoracic aortic organ bath studies were performed on tissues from 8-week-old healthy Wistar rats, using varying concentrations of C. gigas oyster extract. To elucidate a mechanism of action for the oyster's vasoactive properties, concentration response curves were carried out in the presence of a calcium channel inhibitior (verapamil), a nitric oxide synthase inhibitor (N(G)-nitro-L-arginine methyl ester), a potassium channel inhibitor (4-aminopyridine), in addition to the α-adrenoceptor inhibitor prazosin. RESULTS: Oyster solution at 7 500 mg/mL inhibited noradrenaline-induced contraction in isolated aortic rings. Cardiac electrophysiology results showed that neither concentration of oyster solution was able to significantly reduce action potential duration at all phases of repolarisation in left ventricular papillary muscles from healthy animals. CONCLUSION: When administered to healthy vascular tissue, C. gigas oyster extract inhibits contraction induced by noradrenaline. This effect is likely to be mediated through α-adrenoceptor inhibition, and to a lesser extent, calcium modulating activity.

Temperature dependence of Congo red binding to amyloid ß12-28.

Because Congo red (CR) can bind to critical intermediate structural forms of amyloid beta (Aß), it has been suggested as a potential therapeutic agent against neurodegenerative disorders such as Alzheimer's disease. In this study, the interaction of CR with Aß(12-28) was investigated by use of isothermal titration calorimetry (ITC). Studies conducted between 15 and 35 °C show that binding of CR to Aß(12-28) was strongly dependent on temperature, with a decrease in CR-Aß(12-28) complexation as temperature increases, presumably because of conformational changes within Aß(12-28) at the highest temperatures, that conceal the CR binding sites. In fact, no CR binding was observed at 35 °C. The binding of CR to Aß(12-28) was associated with favorable changes in both enthalpy and entropy that resulted in binding constants (K) of between 10(5) and 10(6) M (-1). An early (and more intense) entropy-driven CR disaggregation phase (K ~10(7)-10(8) M (-1)) was observed before the onset of CR-Aß(12-28) complexation. Only CR disaggregation was observed at 35 °C. These results may provide further insights into the ability of CR to inhibit Aß toxicity in neurodegenerative diseases.

Simulations of a lattice model of two-headed linear amphiphiles: influence of amphiphile asymmetry.

Using a 2D lattice model, we conduct Monte Carlo simulations of micellar aggregation of linear-chain amphiphiles having two solvophilic head groups. In the context of this simple model, we quantify how the amphiphile architecture influences the critical micelle concentration (CMC), with a particular focus on the role of the asymmetry of the amphiphile structure. Accordingly, we study all possible arrangements of the head groups along amphiphile chains of fixed length N = 12 and 16 molecular units. This set of idealized amphiphile architectures approximates many cases of symmetric and asymmetric gemini surfactants, double-headed surfactants, and boloform surfactants. Consistent with earlier results, we find that the number of spacer units s separating the heads has a significant influence on the CMC, with the CMC increasing with s for s < N/2. In comparison, the influence of the asymmetry of the chain architecture on the CMC is much weaker, as is also found experimentally.

Fully integrated three-dimensional electrodes for electrochemical detection in microchips: fabrication, characterization, and applications.

A scalable and rather inexpensive solution to producing microanalytical systems with "on-chip" three-dimensional (3D) microelectrodes is presented in this study, along with applicability to practical electrochemical (EC) detection scenarios such as preconcentration and interferant removal. This technique to create high-aspect-ratio (as much as 4:1) gold microstructures in constrained areas involved the modification of stud bump geometry with microfabricated silicon molds via an optimized combination of temperature, pressure, and time. The microelectrodes that resulted consisted of an array of square pillars approximately 18 microm tall and 20 microm wide on each side, placed at the end of a microfabricated electrophoresis channel. This technique increased the active surface area of the microelectrodes by as much as a factor of 50, while mass transfer and, consequently, preconcentration collection efficiencies were increased to approximately 100%, compared to approximately 30% efficiency for planar nonmodified microelectrodes (samples that were used included the neurotransmitters dopamine and catechol). The 3D microelectrodes were used both in a stand-alone configuration, for direct EC detection of model catecholamine analytes, and, more interestingly, in dual electrode configurations for EC sample processing prior to detection downstream at a second planar electrode. In particular, the 3D electrodes were shown to be capable of performing coulometry or complete (100%) redox conversion of analyte species over a wide range of concentrations, from 4.3 microM to 4.4 mM, in either plug-flow or continuous-flow formats.

History of computer-assisted orthopedic surgery (CAOS) in sports medicine.

Computer-assisted orthopedic surgery and navigation applications have a history rooted in the desire to link imaging technology with real-time anatomic landmarks. Although applications are still evolving in the clinical and research setting, computer-assisted orthopedic surgery has already demonstrated in certain procedures its potential for improving the surgeon's accuracy, reproducibility (once past the learning curve), and in reducing outlier outcomes. It is also being used as an educational tool to assist less experienced surgeons in interpreting measurements and precision placements related to well defined anatomic landmarks. It also can assist experienced surgeons, in real-time, plan their bony cuts, tunnel placement, and with ligament balancing. Presently, the additional time, the expense to acquire the needed software and hardware, and restricted reimbursement have slowed the widespread use of navigation. Its current applications have been primarily in joint replacement surgery, spine surgery, and trauma. It has not been widely used in the clinical setting for sports medicine procedures. Sports medicine applications such as individualizing tunnel placement in ligament surgery, opening wedge osteotomy with and without accompanying ligament reconstruction, and balancing and tensioning of the ligaments during the procedure (allowing real-time corrections if necessary) are currently being evaluated and being used on a limited clinical basis.

Ten-year magnetic resonance imaging follow-up of bioabsorbable poly-L-lactic acid interference screws after anterior cruciate ligament reconstruction.

In a previous study bioabsorbable screws composed of poly-L-lactic acid were used for graft fixation and examined prospectively with serial magnetic resonance imaging (MRI) scans 8, 16, and 24 months after anterior cruciate ligament reconstruction. Autogenous patellar tendon graft was used in all 10 patients. Although the manufacturer's estimated time to absorption was 2 years, only 1 of 20 screws had been absorbed at 24 months' follow-up. After 10 years, 6 patients were available for follow-up. None of the patients had instability, persistent effusions, or clinically detectable adverse reactions to the screws. MRI showed that all of the screws had been absorbed. However, all of the patients who underwent MRI had evidence of intraosseous fluid collections at the tibial BioScrew site (Linvatec, Largo, FL), and 4 of 6 had fluid collections at the femoral screw site. The persistence of intraosseous fluid collections 10 years after implantation underscores the importance of long-term follow-up of bioabsorbable implants, particularly given the rapid evolution of bioabsorbable materials used for interference screws.

Sevoflurane provides earlier tracheal extubation and assessment of cognitive recovery than isoflurane in patients undergoing off-pump coronary artery bypass surgery.

OBJECTIVE: To determine whether sevoflurane, because of its lower blood/gas partition coefficient, compared with isoflurane as the primary anesthetic agent, allows earlier tracheal extubation and assessment of cognitive function after off-pump coronary artery bypass (OPCAB) surgery. DESIGN: Prospectively, patients were randomly assigned to receive sevoflurane or isoflurane as their primary anesthetic. Intraoperative opioids were limited to 5 microg/kg of fentanyl. SETTING: Two university hospitals with active cardiac surgery programs. PARTICIPANTS: One hundred one OPCAB surgery patients who met inclusionary and exclusionary criteria participated with institutional review board approval. INTERVENTIONS: Mini-Mental Status Examination, Memory Recall Test, and Observer Assessment of Anxiety and Sedation scales were administered preoperatively, postextubation, at 90 minutes, and between 12 to 24 hours. Pain scores were obtained every 15 minutes after extubation for 90 minutes. MEASUREMENTS AND MAIN RESULTS: Sevoflurane patients were extubated earlier than isoflurane patients (Sevo, 176 +/- 217 minutes and Iso, 257 +/- 279 min, p = 0.02). Although both agents produced similar postanesthetic cognitive profiles, cognitive testing occurred approximately 90 minutes earlier in the sevoflurane group. Verbal rating scale for pain scores >5 were more frequent for sevoflurane than isoflurane patients (p = 0.03). CONCLUSIONS: Both sevoflurane and isoflurane may be safely used as maintenance agents in OPCAB. Sevoflurane has the advantage of allowing earlier extubation and evaluation of cognitive and neurologic function after OPCAB.

Technical aspects of computer-assisted opening wedge high tibial osteotomy.

This study reports a technical approach used in the application of computer-assisted navigation for patients undergoing a medial opening wedge proximal tibial osteotomy. Alignment of the limb can be monitored and documented at the beginning, during, and end of surgery. Surgical incisions are 4-6 cm in length and operative time (including the intra-articular arthroscopic procedure and osteotomy) averages 68 minutes, with 12 seconds of total fluoroscopy using a mini C-arm. Follow-up radiographic assessment in the office setting confirms corrections within 2 degrees of the planned correction. Computer-assisted navigation is a valuable adjunct during opening wedge proximal tibial osteotomy as it provides real-time intraoperative information that contributes to and confirms decision-making.

Ion-molecule reactions of several ions with ethylene oxide and propenal in a selected ion flow tube.

The selected ion flow tube (SIFT) technique has been used to investigate the ion-molecule reactions of several ions with the neutral molecules ethylene oxide, CH(2)OCH(2)-c, and propenal, CH(2)CHCHO. Both molecules have been identified in hot-core star forming regions [] and have significance to astrochemical models of the interstellar (ISM) and circumstellar medium (CSM). Moreover, the molecules contain functional groups, such as the epoxide group (ethylene oxide) and an aldehyde group, which are part of a conjugated pi-electron system (propenal) whose reactivities have not been studied in detail in gas-phase ion-molecule reactions. The larger recombination energy ions, Ar(+) and N(2)(+), were reacted with the neutrals to give insight into general fragmentation tendencies. These reactions proceeded via dissociative charge-transfer yielding major fragmentation products of CH(3)(+) and HCO(+) for ethylene oxide and CH(2)CH(+) and HCO(+) for propenal. The amino acids glycine and alanine are of particular interest to astrobiology, especially if they can be synthesized in the gas phase. In an attempt to synthesize amino acid precursors, ethylene oxide and propenal were reacted with NH(n)(+) (n = 1-4) and HCNH(+). As might be expected from the proton detachment energies, NH(+), NH(2)(+), and HCNH(+) reacted via proton transfer. NH(3)(+) reacted with each molecule via H-atom abstraction to produce NH(4)(+), and NH(4)(+) reacted via a ternary association. All binary reactions proceeded near the gas kinetic rate. Several associated molecule switching reactions were performed and implications of these reactions to the structures of the association products are discussed Ikeda et al. and Hollis et al.

Articular cartilage: injury pathways and treatment options.

Articular cartilage injury and degeneration is a frequent occurrence in synovial joints. Treatment of these articular cartilage lesions are a challenge because this tissue is incapable of quality repair and/or regeneration to its native state. Nonoperative treatments endeavor to control symptoms, and include anti-inflammatory medication, viscosupplementation, bracing, orthotics, and activity modification. Techniques to stimulate the intrinsic repair (fibrocartilage) process include drilling, abrasion, and microfracture of the subchondral bone. Currently, the clinical biologic approaches to treat cartilage defects include autologous chondrocyte implantation, periosteal transfer, and osteochondral autograft or allograft transplantation. Newer strategies employing tissue engineering being studied involve the use of combinations of progenitor cells, bioactive factors, and matrices, and the use of focal synthetic devices. Many new and innovative treatments are being explored in this exciting field. However, there is a paucity of prospective, randomized controlled clinical trials that have compared the various techniques, treatment options, indications and efficacy.

Fabrication of a glass capillary electrophoresis microchip with integrated electrodes.

In this chapter, a detailed outline delineating the processing steps for microfabricating capillary electrophoresis (CE) with integrated electrochemical detection (ECD) platforms for performing analyte separation and detection is presented to enable persons familiar with microfabrication to enter a cleanroom and fabricate a fully functional Lab-on-a-Chip (LOC) microdevice. The processing steps outlined are appropriate for the production of LOC prototypes using easily obtained glass substrates and common microfabrication techniques. Microfabrication provides a major advantage over existing macro-scale systems by enabling precise control over electrode placement, and integration of all required CE and ECD electrodes directly onto a single substrate with a small footprint. In the processing sequences presented, top and bottom glass substrates are photolithographically patterned and etched using wet chemical processing techniques. The bottom substrate contains seven electrodes required for CE/ECD operation, whereas the top substrate contains the microchannel network. The flush planar electrodes are created using sputter deposition and lift-off processing techniques. Finally, the two glass substrates are thermally bonded to create the final LOC device.