Integrated Microfluidic System for Rapid DNA Fingerprint Analysis: A Miniaturized Integrated DNA Analysis System (MiDAS)-Swab Sample-In to DNA Profile-Out.

A fully automated rapid DNA analysis system requires integrating several operational elements performing multiple steps onto one single microfluidic platform. The functions to include on the microfluidic platform consist of substrate lysis, lysate DNA extraction, single or multiplexed PCR amplification, amplicon separation, and product readout. Here we describe a fully automated integrated system for forensic short tandem repeat (STR) analysis of reference samples, achieving buccal swab-in and DNA profile-out.

Optimization of multiplexed PCR on an integrated microfluidic forensic platform for rapid DNA analysis.

This study reports the design, prototyping, and assay development of multiplexed polymerase chain reaction (PCR) on a plastic microfluidic device. Amplification of 17 DNA loci is carried out directly on-chip as part of a system for continuous workflow processing from sample preparation (SP) to capillary electrophoresis (CE). For enhanced performance of on-chip PCR amplification, improved control systems have been developed making use of customized Peltier assemblies, valve actuators, software, and amplification chemistry protocols.

Gait and balance in treatment-naïve active alcoholics with and without a lifetime drug codependence.

Background: Disturbed gait and balance are among the most consistent sequelae of chronic alcoholism. However, although a majority of alcoholics have never sought treatment, most investigations showing ataxia in alcohol-dependent individuals have relied on samples drawn from treated populations. In addition, few studies have addressed the associations of codependence on other drugs with alcoholic gait and balance disturbance.

Persistent but less severe ataxia in long-term versus short-term abstinent alcoholic men and women: a cross-sectional analysis.

Background: Disturbed gait and balance are among the most consistent and salient sequelae of chronic alcoholism. Results of small sample longitudinal investigations have provided evidence that partial recovery of gait and balance functions in alcoholics may be achieved with abstinence. However, abstinence durations reported have been limited, and their power and generalizability have suffered from small sample sizes.

Subcortical volumes in long-term abstinent alcoholics: associations with psychiatric comorbidity.

Background: Research in chronic alcoholics on memory, decision-making, learning, stress, and reward circuitry has increasingly highlighted the importance of subcortical brain structures. In addition, epidemiological studies have established the pervasiveness of co-occurring psychiatric diagnoses in alcoholism. Subcortical structures have been implicated in externalizing pathology, including alcohol dependence, and in dysregulated stress and reward circuitry in anxiety and mood disorders and alcohol dependence.

Cognitive performance in treatment-naïve active alcoholics.

Background: Most studies reporting cognitive deficits in chronic alcoholics have relied on treatment samples (predominantly men) from inpatient or outpatient treatment facilities. However, the majority of chronic alcoholics have never been in treatment and there is increasing evidence that treated and non-treatment-seeking alcoholic samples come from different populations with regard to alcohol use and other factors related to the severity of disease. Accordingly, in the present study, we assessed a broad range of cognitive functions in 55 treatment-naïve alcohol-dependent (TNAD) individuals and 55 nonalcoholic controls (NAC) matched for age and education.

Integrated microfluidic system for rapid forensic DNA analysis: sample collection to DNA profile.

We demonstrate a conduit for the delivery of a step change in the DNA analysis process: A fully integrated instrument for the analysis of multiplex short tandem repeat DNA profiles from reference buccal samples is described and is suitable for the processing of such samples within a forensic environment such as a police custody suite or booking office. The instrument is loaded with a DNA processing cartridge which incorporates on-board pumps and valves which direct the delivery of sample and reagents to the various reaction chambers to allow DNA purification, amplification of the DNA by PCR, and collection of the amplified product for delivery to an integral CE chip. The fluorescently labeled product is separated using micro capillary electrophoresis with a resolution of 1.

Cyclic 3', 5' guanosine monophosphate and larval midgut ecdysone 20-monooxygenase activity of the tobacco hornworm, Manduca sexta.

During the last larval stadium of the tobacco hornworm, Manduca sexta, midgut ecdysone 20-monooxygenase (E20M) activity increases 50-fold coincident with wandering onset. In our study, the effects of the ecdysone agonist RH-5849 and second messengers, dibutyryl cyclic 3', 5' adenosine monophosphate (dibutyryl cAMP) and dibutyryl cyclic 3', 5' guanosine monophosphate (dibutyryl cGMP), either alone or combined with other additives, on midgut E20M were assessed in vitro and in vivo. Pre-incubation of midguts with dibutyryl cAMP was ineffective, but dibutyryl cGMP elevated E20M activity.

Feasibility of Internet training for care staff of residents with dementia: the CARES program.

This study evaluated an Internet-based training module, CARES(®) (Connect with the resident; Assess behavior; Respond appropriately; Evaluate what works; Share with the team), to determine its feasibility for certified nursing assistants (CNAs) in three nursing homes and one assisted living facility. Pre- and posttest questionnaires were administered to 40 CNAs to determine improvements in dementia care knowledge and perceptions of competence in dementia care. Dementia care knowledge improved significantly after CARES training.

Midgut and fatbody mitochondrial transhydrogenase activities during larval-pupal development of the tobacco hornworm, Manduca sexta.

Midgut and fatbody mitochondria from fifth larval instar Manduca sexta display a membrane-associated transhydrogenase that catalyzes a reversible hydride ion transfer between NADP(H) and NAD(H). The NADPH-forming activity occurs as a nonenergy- or energy-linked activity with energy for the latter derived from either electron transport-dependent NADH or succinate utilization, or ATP hydrolysis by Mg(++)-dependent ATPase. During the ten-day developmental period preceding the larval-pupal molt (fifth larval instar), significant peaks in the mitochondrial transhydrogenase activities of midgut and fatbody tissues were noted and these peaks were coincident with the onset of wandering behavior and with the fifty-fold increase in ecdysone 20-monooxygenase (E20-M) activity previously reported for M.

Midgut mitochondrial transhydrogenase in wandering stage larvae of the tobacco hornworm, Manduca sexta.

Midgut mitochondria from fifth larval instar Manduca sexta exhibited a transhydrogenase that catalyzes the following reversible reaction: NADPH + NAD(+) <--> NADP(+) + NADH. The NADPH-forming transhydrogenation occurred as a nonenergy- and energy-linked activity. Energy for the latter was derived from the electron transport-dependent utilization of NADH or succinate, or from Mg++-dependent ATP hydrolysis by ATPase.

Effects of the compounds 2-methoxynaphthoquinone, 2-propoxynaphthoquinone, and 2-isopropoxynaphthoquinone on ecdysone 20-monooxygenase activity.

The effects of the natural compound 2-methoxy-1,4-naphthoquinone, isolated from the leaves of Impatiens glandulifera and the synthetic compounds 2-propoxy-1,4-naphthoquinone and 2-isopropoxy-1,4-naphthoquinone on ecdysone 20-monooxygenase (E-20-M) activity were examined in three insect species. Homogenates of wandering stage third instar larvae of Drosophila melanogaster, or abdomens from adult female Aedes aegypti, or fat body or midgut from fifth instar larvae of Manduca sexta were incubated with radiolabelled ecdysone and increasing concentrations (from 1 x 10(-8) to 1 x 10(-3) M) of the three compounds. All three compounds were found to inhibit in a dose-dependent fashion the E-20-M activity in the three insect species.