bluuebunny/crossref_metadata_2025_split huggingface dataset:
5-Hydroxytryptamine Type 3 Receptor Modulates Opioid-induced Hyperalgesia and Tolerance in Mice
Authors: Liang De-Yong , Li XiangQi , Clark J. David 
Published: March 2011

Background
Opioid-induced hyperalgesia (OIH) and tolerance are challenging maladaptations associated with opioids in managing pain. Recent genetic studies and the existing literature suggest the 5-hydroxytryptamine type 3 (5-HT3) receptor participates in these phenomena. The location of the relevant receptor populations and the interactions between the 5-HT3 system and other systems controlling OIH and tolerance have not been explored, however. We hypothesized that 5-HT3 receptors modulate OIH and tolerance, and that this modulation involves the control of expression of multiple neurotransmitter and receptor systems.

Methods
C57BL/6 mice were exposed to a standardized 4-day morphine administration protocol. The 5-HT3 antagonist ondansetron was administered either during or after the conclusion of morphine administration. Mechanical testing was used to quantify OIH, and thermal tail-flick responses were used to measure morphine tolerance. In other experiments spinal cord and dorsal root ganglion tissues were harvested for analysis of messenger RNA concentrations by real-time polymerase chain reaction or immunochemistry analysis.

Results
The results showed that (1) systemic or intrathecal injection of ondansetron significantly prevented and reversed OIH, but not local intraplantar injection; (2) systemic or intrathecal injection of ondansetron prevented and reversed tolerance; and (3) ondansetron blocked morphine-induced increases of multiple genes relevant to OIH and tolerance in dorsal root ganglion and spinal cord.

Conclusions
Morphine acts via a 5-HT3-dependent mechanism to support multiple maladaptations to the chronic administration of morphine. Furthermore, the use of 5-HT3 receptor antagonists may provide a new avenue to prevent or reverse OIH and tolerance associated with chronic opioid use.
Recording Sound-Evoked Potentials from the Drosophila Antennal Nerve: Figure 1.
Authors: Eberl Daniel F. , Kernan Maurice J. 
Published: March 2011

INTRODUCTIONInformation about an animal's social and ecological environment takes many forms, several of which can be sensed as mechanical signals. The mechanosensory world of a fly such as Drosophila melanogaster is especially complex, and the transduction of signals involves a diverse set of specialized sense organs. Near-field sounds, such as the male courtship song, are transduced by sensory units in Johnston's organ (JO), which is the chordotonal organ in the Drosophila antennae. This article describes a protocol for recording extracellular potentials in the antennal nerve that occur in response to near-field acoustic signals generated electronically to mimic courtship songs. These recordings represent the combined action potentials of many or perhaps even all of the responding JO sensory neurons whose axons project along the antennal nerve en route to the antennal mechanosensory and motor center (AMMC).
Preparation of Meiotic Chromosomes from Larval and Pupal Drosophila Testes
Authors: Bonaccorsi Silvia , Giansanti Maria G. , Cenci Giovanni , Gatti Maurizio 
Published: March 2011

INTRODUCTIONSeveral techniques have been developed for the preparation of Drosophila male meiotic chromosomes. All of these techniques allow a clear visualization of chromosomes, but not of other cellular structures such as the cytoskeleton and the spindle. This protocol describes two methods for preparation of meiotic chromosomes from larval and pupal testes. The first is a very simple method for preparing aceto-orcein-stained chromosomes. The second allows preparation of unstained chromosomes that can subsequently be stained with various dyes such as Giemsa, Hoechst 33258, and DAPI, or processed for in situ hybridization.
Micropatterning Cell-Substrate Adhesions Using Linear Polyacrylamide as the Blocking Agent
Authors: Guo Wei-hui , Wang Yu-li 
Published: March 2011

INTRODUCTIONRecent developments in micropatterning techniques have facilitated systematic investigations of how cells respond to geometric cues, which likely have an important role in many physiological processes, such as growth and differentiation. Micropatterned substrates also offer technical advantages for cell biological studies by normalizing cell shapes and reducing the variability among cultured cells. In this article, we present a simple, economical micropatterning method that may be easily implemented in most laboratories. We also describe how to treat the substrate with linear polyacrylamide (PAA) to inhibit unwanted cell adhesion. First, glass coverslip surfaces are activated with 3-methacryloxypropyltrimethoxysilane (bind-silane), an approach that was originally used for covalently linking PAA gels to glass plates for SDS-PAGE to prevent gels from shrinking and swelling during staining/destaining. The activated surfaces are then coated with the photoresist SPR-220, exposed to ultraviolet (UV) light through a photomask, and developed to selectively remove UV-exposed photoresist. Finally, linear PAA is grafted onto the exposed areas of the bind-silane substrate (to prevent cell adhesion), and the remaining photoresist is removed to uncover the glass surface, making the unexposed areas available as adhesive areas for cell culture.
Dendrimer-Mediated Transfection
Authors: Dean David A. , Gasiorowski Joshua Z. 
Published: March 2011

INTRODUCTIONThe efficient delivery of exogenous DNA to cells for expression and function studies is an essential technique of modern cell biology. Polymer-based systems can be used to transfect cells, some of which rely on one type of molecule, whereas others contain a mixture of polymers and lipids. Reagents such as SuperFect or PolyFect use activated dendrimers, which are positively charged and resemble snowflakes in structure. One advantage of using dendrimer-based reagents is that they appear to be less sensitive than liposomal reagents to the presence of serum during transfection. Thus, they could be a better choice for cells that show greater dependence on the continual presence of serum (to avoid initiation of stress responses or induction of differentiation). This protocol describes a procedure for dendrimer-mediated transfection.
Preparing Injection Pipettes on a Flaming/Brown Pipette Puller
Authors: Dean David A. , Gasiorowski Joshua Z. 
Published: March 2011

INTRODUCTIONThe efficient delivery of exogenous DNA to cells for expression and function studies is an essential technique of modern cell biology, and direct delivery of genetic material by microinjection remains a reliable means of transfection. Needles for microinjection can either be pulled from glass capillaries on a pipette puller or can be purchased premade. When pulling needles, variables such as filament design, heat, pull strength (tension), and delay time between heating and pulling must be addressed. The heat setting affects the length and the tip size of the needle; high heat will typically produce longer needles and finer tips. The pull strength will also affect length and tip size, with greater pull strength producing longer tapered needles with finer tips. Finally, shorter delay times between heating and pulling can result in longer tapers and finer needles. However, if the delay is too short, the glass forms fibers resembling glass wool instead of usable needles. The advantage of pulling needles in the laboratory is that a variety of different needle types can be pulled, depending on the samples and the cells being injected. For laboratories performing even a moderate number of microinjection experiments, a Flaming/Brown-style pipette puller such as the Sutter Instruments P-97 is a cost-effective choice for manufacturing quality micropipettes quickly and reproducibly.
Preparing Injection Pipettes on a PUL-1 Micropipette Puller
Authors: Dean David A. , Gasiorowski Joshua Z. 
Published: March 2011

INTRODUCTIONThe efficient delivery of exogenous DNA to cells for expression and function studies is an essential technique of modern cell biology, and direct delivery of genetic material by microinjection remains a reliable means of transfection. Needles for microinjection can either be pulled from glass capillaries on a pipette puller or be purchased premade. When pulling needles, variables such as filament design, heat, pull strength (tension), and delay time between heating and pulling must be addressed. The heat setting affects the length and the tip size of the needle; high heat will typically produce longer needles and finer tips. The pull strength will also affect length and tip size, with greater pull strength producing longer tapered needles with finer tips. Finally, shorter delay times between heating and pulling can result in longer tapers and finer needles; if the delay is too short, the glass forms fibers resembling glass wool rather than usable needles. The advantage of pulling needles in the laboratory is that a variety of different needle types can be pulled, depending on the samples and the cells being injected. The PUL-1 micropipette puller is a robust and inexpensive machine that can be found in many laboratories around the world. New machines are no longer being manufactured, although used ones can still be purchased.
Differential Detergent Fractionation of Eukaryotic Cells
Authors: Ramsby Melinda , Makowski Gregory 
Published: March 2011

INTRODUCTIONDifferential detergent fractionation (DDF) involves the sequential extraction of cells with PIPES buffers containing first digitonin, then Triton, and finally Tween/deoxycholate (DOC). The procedure yields four biochemically and electrophoretically distinct fractions composed of the following: (1) cytosolic proteins and extractable cytoskeletal elements; (2) membrane and organelle proteins; (3) nuclear membrane proteins and extractable nuclear proteins; (4) detergent-resistant cytoskeletal filaments and nuclear matrix proteins. Most of these fractions can be enriched or purified further by subsequent manipulations. The DDF protocol described here represents a modification of a method used for the fractionation of Madin-Darby canine kidney cells, including the addition of a digitonin extraction step, the inclusion of EDTA in the digitonin and Triton buffers, and the elimination of a nuclease digestion step; DNA is instead denatured by shear force in the presence of sodium dodecyl sulfate (SDS).
Microbial Opsins: A Family of Single-Component Tools for Optical Control of Neural Activity
Authors: Yizhar Ofer , Fenno Lief , Zhang Feng , Hegemann Peter , Diesseroth Karl 
Published: March 2011

INTRODUCTIONMembers of the microbial opsin gene family have emerged recently as crucial tools for “optogenetics,” a new neuroscience technology. “Optogenetics” can be defined as the integration of optics and genetics to control well-defined events (such as action potentials) within specified cells (such as a targeted class of projection neurons) in living tissues (such as the brains of freely behaving mammals). In this article, we focus on the diversity of the microbial opsin genes and the structure–function properties of their corresponding proteins.
A systematic and methodological review of interventions for young people experiencing alcohol‐related harm
Authors: Calabria Bianca , Shakeshaft Anthony P. , Havard Alys 
Published: March 2011

ABSTRACTAims  This review identified published studies evaluating interventions delivered outside educational settings, designed for young people with existing alcohol use problems, or who participate in behaviour that places them at high risk of alcohol‐related harm, critiqued their methodology and identified opportunities for new interventions.Methods  A systematic search of the peer‐reviewed literature interrogated 10 electronic databases using specific search strings, limited to 2005–09. No additional studies were found by a librarian searching other collections and clearing‐houses, or by hand‐searching review paper reference lists. The 1697 articles identified were reviewed against criteria from the Dictionary for the Effective Public Health Practice Project Quality Assessment Tool for Quantitative Studies.Results  The methodological quality of existing studies is variable, and needs to be both more rigorous and more consistent. Particular problems include the lack of blinding outcome assessors, a reliance solely on self‐report measures, highly variable consent and follow‐up rates, infrequent use of intention‐to‐treat analyses and the absence of any economic or cost analyses. The range of interventions evaluated is currently limited to individually focused approaches, almost exclusively implemented in the United States.Conclusions  There is a great need for more intervention trials for young people at high risk of experiencing alcohol‐related harm that are both methodologically rigorous and have a broader community focus, to complement the psychological interventions that currently dominate the relevant literature. Such trials would improve outcomes for high‐risk young people themselves and would improve the evidence base, both in their own right and by facilitating future meta‐analyses.
Does improved access and greater choice of nicotine replacement therapy affect smoking cessation success? Findings from a randomized controlled trial
Authors: Walker Natalie , Howe Colin , Bullen Chris , Grigg Michele , Glover Marewa , McRobbie Hayden , Laugesen Murray , Jiang Joy , Chen Mei‐Hua , Whittaker Robyn , Rodgers Anthony 
Published: March 2011

ABSTRACTAims  To determine the effect of offering smokers who want to quit easy access to nicotine replacement therapy (NRT), a period of familiarization and choice of product on smoking abstinence at 6 months.Design  Single‐blind, randomized controlled trial.Setting  New Zealand.Participants  A total of 1410 adult smokers who called the national Quitline for quitting support were randomized to usual Quitline care or a box containing different NRT products (patch, gum, inhaler, sublingual tablet, oral pouch) to try for a week prior to quitting, and then to choose one or two of these products for 8 weeks' use.Measurements  The primary outcome was 7‐day point prevalence smoking abstinence 6 months after quit day. Secondary outcomes included continuous abstinence, cigarette consumption, withdrawal, NRT choice and serious adverse events at 1 and 3 weeks and 3 and 6 months.Findings  No differences in 6‐month quit rates (7‐day point prevalence or continuous abstinence) were observed between the groups. However, smokers allocated to the intervention group were more likely to have quit smoking at 3 months [self‐reported point prevalence, relative risk (RR) = 1.17, 95% confidence interval (CI): 1.02, 1.35, P = 0.03], had a longer time to relapse (median 70 days versus 28 days, P < 0.01) and used significantly more NRT. The selection box concept was highly acceptable to users, with the patch and inhaler combination the most popular choice (34%).Conclusions  In terms of smoking abstinence at 6 months, offering smokers who want to quit free access to a wide range of nicotine replacement therapy, including a 1‐week period of familiarization and choice of up to two products, appears no different to offering reduced cost and choice of nicotine replacement therapy, with no familiarization period.
The risk of coronary thrombosis with cyclo‐oxygenase‐2 inhibitors does not vary with polymorphisms in two regions of the cyclo‐oxygenase‐2 gene
Authors: McGettigan Patricia , Lincz Lisa F. , Attia John , McElduff Patrick , Bissett Linda , Peel Roseanne , Stokes Barrie , Hancock Stephen , Henderson Kim , Seldon Michael , Henry David 
Published: March 2011

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT• The risk of cardiovascular events with non‐steroidal anti‐inflammatory drugs (NSAIDs) varies with individual drugs and with dosage. Little is known about other factors that can modify risk.• Polymorphisms of the COX‐2 gene conceivably could alter the risk of myocardial infarction and also the risk from NSAIDs.WHAT THIS STUDY ADDS• Two common polymorphisms of the COX‐2 gene, rs 20417 (G > C) and rs 5275 (T > C), were not associated with variation in the risk of acute coronary syndrome, and individually did not alter the risk associated with use of NSAIDs.• Haplotype analysis identified a trend to protection against NSAID‐related cardiovascular risk with the ‘low risk’ haplotype, but the effect was weak and if confirmed, the clinical utility would be limited.AIMS To investigate whether polymorphisms of the cyclo‐oxygenase‐2 (COX‐2) gene modify the adverse cardiovascular effects of COX‐2 inhibitors.METHODS A case control study was conducted in the Hunter Region of New South Wales, Australia. Cases (n= 460) were hospitalized with acute coronary syndrome (ACS). Controls (n= 640) were recruited from the electoral rolls. Structured interviews gathered information on variables including recent ingestion of non‐steroidal anti‐inflammatory drugs (NSAIDs). Targeted genotyping of rs 20417(G > C) and rs5275 (T > C) polymorphisms was performed by real‐time polymerase chain reaction using allele‐specific probesRESULTS Ingestion of any NSAID in the week prior to interview was associated with an elevated risk for ACS: adjusted odds ratio 1.8 (1.2, 2.5). The rs 20417 and rs 5275 polymorphisms were not singly associated with risk for ACS: adjusted odds ratios 1.1 (0.80, 1.5) and 1.2 (0.88, 1.5), respectively. Individually, the polymorphisms did not modify the risk of ACS with the drugs. When analyses were conducted by haplotype, the adjusted odds ratio with celecoxib or rofecoxib in individuals who had one or two copies of the ‘low risk’ haplotype (no GT) was 1.2 (0.29, 5.0), compared with 2.1 (1.1, 4.0) with the ‘high risk’ haplotype (one or two copies of GT).CONCLUSIONS We found little evidence of a gene/drug interaction. We found a statistically non‐significant trend toward a lower risk of coronary events with NSAIDs in the presence of the ‘low risk’ haplotype. Even if confirmed, the clinical utility of the finding would be limited as this haplotype is carried by a minority of the population.
Irritability: an underappreciated side effect of interferon treatment for chronic hepatitis C?
Authors: Blacklaws Helen , Gardner Anne , Usher Kim 
Published: March 2011

Aim and objectives.  The research literature was reviewed with the aim of answering the question ‘is irritability an underappreciated side effect of interferon and ribavirin treatment for hepatitis C’.Background.  The majority of information regarding interferon treatment identifies depression as the main psychological side effect. However, clinical observation and patient reports suggest that irritability, not depression, is the predominant side effect.Design.  The literature review included research and discussion papers. Data bases were searched using the keywords interferon and hepatitis C in combination with one of the following: side effects, depression, mood alteration/change, irritability, anger, impulse control, psychiatric side effects or neuropsychiatric side effects.Results.  The review revealed a gap in the literature regarding interferon‐related irritability. Whereas depression was well researched and described, irritability was afforded little research time. However, where irritability was assessed, it was found to occur to a significant degree. Issues identified were difficulty defining and categorising irritability; lack of irritability‐specific assessment tools and failure of depression rating scales to adequately discern irritable mood; and the confounding effect of physiological side effects on mood alteration.Relevance to clinical practice.  Underappreciation and underrecognition of irritability have implications for clinical practice. Good research is the foundation for evidence‐based practice; therefore, the possibility exists that, based on current research evidence, patients may not be receiving a standard care that adequately addresses the entirety of the side effect spectrum.Conclusion.  Irritability is an underappreciated psychological side effect of interferon therapy. Although irritability is recognised as a side effect of interferon, there is considerable discordance between clinical observation, patient reports and research evidence as reported in the literature.
Adult sibling experience, roles, relationships and future concerns – a review of the literature in learning disabilities
Authors: Davys Deborah , Mitchell Duncan , Haigh Carol 
Published: March 2011

Aim.  This paper provides a review of the literature related to adult siblings of learning‐disabled people.Background.  Siblings of learning‐disabled people are often looked upon as next of kin when older parents die; however, there is little research regarding sibling views and wishes.Design.  A literature review of published peer‐reviewed empirical research was undertaken.Method.  Electronic databases and citation tracking were used to collate data using key terms such as adult siblings and learning disability. Relevant articles were analysed, compared and contrasted.Results.  Six key themes emerged suggesting a varied impact of learning disability upon sibling lives in areas that include life choices, relationships, identity and future plans. Some siblings report a positive impact upon life, others state their lives are comparable with other adults who do not have a learning‐disabled sibling and others still report a negative impact. Sibling roles and relationships are varied. Evidence suggests that sibling roles, relationships and experience are affected by life stage. Parents often have a primary care role for the disabled person, whilst siblings perform a more distant role; however, sibling involvement often rises when parents are no longer able to provide previous levels of support.Conclusion.  Many factors appear to affect the sibling experience and uptake of roles including gender, life stage and circumstances, level of disability, health status and relationships between family members. Siblings are concerned about the future, particularly when parents are no longer able to provide support, and many appear to have expectations of future responsibilities regarding their disabled sibling.Relevance to practice.  As siblings of people who have a learning disability are often expected by society to provide support, it is important that health and social care practitioners are aware of issues that may impact on this relationship.
Pharmacokinetics of tulathromycin after single and multiple subcutaneous injections in domestic goats (Capra aegagrus hircus)
Authors: CLOTHIER K. A. , LEAVENS T. , GRIFFITH R. W. , WETZLICH S. E. , BAYNES R. E. , RIVIERE J. E. , TELL L. A. 
Published: March 2011

Clothier, K. A., Leavens, T., Griffith, R. W., Wetzlich, S. E., Baynes, R. E., Riviere, J. E., Tell, L. A. Pharmacokinetics of tulathromycin after single and multiple subcutaneous injections in domestic goats (Capra aegagrus hircus). J. vet. Pharmacol. Therap.34, 448–454.Tulathromycin, a novel triamilide in the macrolide class, is labeled for treatment of bacterial pneumonia in cattle and swine. This manuscript evaluates pharmacokinetics of tulathromycin in goats. In two different studies, six juvenile and ten market‐age goats received a single injection of 2.5 mg/kg of tulathromycin subcutaneously; in a third study, 18 juvenile goats were treated with 2.5, 7.5, or 12.5 mg/kg tulathromycin weekly with three subcutaneous injections. Pharmacokinetic parameters estimated from the plasma concentrations from single injections were similar between the two groups of goats and to previously reported parameters in cattle and swine. Mean terminal half‐lives were 59.1 ± 7.6 and 61.2 ± 8.7 h for juvenile and market‐age goats, respectively. In the multi‐dose study, pharmacokinetic parameters estimated from plasma concentrations demonstrated significant differences at P < 0.05 among repeated injections but not among doses. Overall, pharmacokinetic parameters in goats are similar to those reported in cattle and swine, and tulathromycin may prove a useful drug for treating respiratory disease in goats.