Implementing a National Alcohol Consumption Prevention Program at the Local Level: What Does Early Evaluation Tell Us?

The current Swedish national plan for preventing alcohol consumption-related problems puts greater emphasis on community-level prevention measures. This report describes early results from the implementation of this plan in the county of Västra Götaland, in southwestern Sweden. During 2002–2004 interviews were conducted with the main project representative in each of the county's 49 municipalities. In addition, more extensive data gathering, involving interviews with a broad range of stakeholders and systematic examination of project-related documentation, was undertaken in four representative municipalities. Findings indicated that community alcohol consumption prevention has been strengthened and that the activity level is high in all municipalities, especially in relation to youth. However, problematic alcoholic beverage consumption by adults was rarely targeted; the limited resources were dispersed over too many projects; there was too little collaboration between stakeholding authorities and adherence to evidence-based practice still is lacking.     

How Does Bacillus thuringiensis Crystallize Such a Large Diversity of Toxins?

Bacillus thuringiensis (Bt) is a natural crystal-making bacterium. Bt diversified into many subspecies that have evolved to produce crystals of hundreds of pesticidal proteins with radically different structures. Their crystalline form ensures stability and controlled release of these major virulence factors. They are responsible for the toxicity and host specificity of Bt, explaining its worldwide use as a biological insecticide. Most research has been devoted to understanding the mechanisms of toxicity of these toxins while the features driving their crystallization have long remained elusive, essentially due to technical limitations. The evolution of methods in structural biology, pushing back the limits in size of amenable protein crystals now allows access to be gained to structural information hidden within natural crystals of such toxins. In this review, we present the main parameters that have been identified as key drivers of toxin crystallization in Bt, notably in the light of recent discoveries driven by structural biology studies. Then, we develop how the future evolution of structural biology will hopefully unveil new mechanisms of Bt toxin crystallization, opening the door to their hijacking with the aim of developing a versatile in vivo crystallization platform of high academic and industrial interest.     

Intravascular Delivery of Particulate Systems: Does Geometry Really Matter?

In cancer therapy and imaging, the systemic passive delivery of particulate systems has relied on the enhanced permeability and retention (EPR) effect: sufficiently small particles can cross the endothelial fenestrations and accumulate in the tumor parenchyma. The vast majority of man-made particulates exhibit a spherical shape as a result of surface energy minimization during their synthesis. The advent of phage display libraries, which are revealing the extraordinary molecular diversity of endothelial cells, and the development of processes for fabricating particles with shapes other than spherical are opening the path to new design solutions for systemically administered targeted particulates. In this paper, the role of particle geometry (i.e., size and shape) is discussed at the tissue and cellular scales. Emphasis is placed on how the synergistic effect of particle geometry and molecular targeting can enhance the specificity of delivery. The intravascular delivery process has been broken into three events: margination, firm adhesion and control of internalization. Predictions from mathematical models and observations from in-vitro experiments were used to show the relevance of particle geometry in systemic delivery. Rational design of particulate systems should consider, beside the physico-chemical properties of the surface coatings, geometrical features as size and shape. The integration of mathematical modeling with in-vitro and in-vivo testing provides the tools for establishing a rational design of nanoparticles.     

Roughness of Production Conditions: Does It Really Affect Stability of IgG-Based Antivenoms?

Antivenoms contain either pure animal IgGs or their fragments as an active substance, and are the only specific therapeutics against envenomation arising from snakebites. Although they are highly needed, the low sustainability of such preparations’ manufacture causes constant global shortages. One reason for this is the stability of the product, which contributes not only to the manufacture sustainability, but the product safety as well. It has been hypothesized that the roughness of conditions to which IgGs are exposed during downstream purification disturbs their conformation, making them prone to aggregation, particularly after exposure to secondary stress. The aim of this research was to investigate how the roughness of the downstream purification conditions influences the stability properties of purified IgGs. For this purpose, equine IgGs were extracted from unique hyperimmune plasma by two mild condition-based operational procedures (anion-exchange chromatography and caprylic acid precipitation) and three rougher ones (ammonium sulphate precipitation, cation-exchange chromatography and protein A affinity chromatography). The stability of the refined preparations was studied under non-optimal storage conditions (37 °C, 42 °C, and a transiently lower pH) by monitoring changes in the aggregate content and thermal stability of the pure IgGs. Mild purification protocols generated IgG samples with a lower aggregate share in comparison to the rougher ones. Their tendency for further aggregation was significantly associated with the initial aggregate share. The thermal stability of IgG molecules and the aggregate content in refined samples were inversely correlated. Since the initial proportion of aggregates in the samples was influenced by the operating conditions, we have shown a strong indication that each of them also indirectly affected the stability of the final preparations. This suggests that mild condition-based refinement protocols indeed generate more stable IgGs.     

Treatment response by primary drug of abuse: Does methamphetamine make a difference?

The purposes of this study were to examine the outcomes of a sample of patients receiving publicly funded substance abuse treatment in Washington State and to compare the outcomes of those using methamphetamine (MA) with patients using other drugs of abuse. All data for this study came from administrative systems in Washington State, and the outcomes included completion of and readmission to treatment, employment, and various forms of criminal justice involvement. Treatment records were linked to outcome data using both deterministic and probabilistic matching techniques. Patients were tracked for 1 year following their discharge, and analyses were performed separately on a study population of adults and a study population of youth. For both adults and youth, the results showed that across outcomes, there were few differences between MA users and users of other hard drugs, whereas there were consistent differences between MA users and users of alcohol and marijuana. Alcohol and marijuana users tended to have more positive outcomes than the other groups. Future research should focus on more detailed analyses of the type of treatment received by patients, particularly for MA users.     

Does a Hot Drink Provide Faster Absorption of Paracetamol Than a Tablet? A Pharmacoscintigraphic Study in Healthy Male Volunteers

Purpose

To investigate the hypothesis that paracetamol is absorbed faster from a hot drink than from a standard tablet using simultaneous scintigraphic imaging and pharmacokinetic sampling.

Methods

Twenty-five healthy male volunteers received both paracetamol formulations in a randomised manner. The formulation administered in the first treatment arm was radiolabelled to allow scintigraphic monitoring. In both treatment arms, blood samples were taken for assessing paracetamol absorption.

Results

Following the hot drink, paracetamol absorption was both significantly faster and greater over the first 60 min post-dose compared with the tablet, as evidenced by the median time to reach t_0.25?μg/mL of 4.6 and 23.1 min, respectively, and AUC_0-60 of 4668.00 and 1331.17 h*ng/mL, respectively. In addition, t_max was significantly shorter for the hot drink (median time = 1.50 h) compared with the tablet (1.99 h). However, C_max was significantly greater following the tablet (9,077 ng/mL) compared with the hot drink (8,062 ng/mL). Onset of gastric emptying after the hot drink was significantly faster than after the standard tablet (7.9 versus 54.2 min), as confirmed scintigraphically.

Conclusions

Compared with a standard tablet, a hot drink provides faster absorption of paracetamol potentially due to more rapid gastric emptying.     

Evaluation of patients’ experiences with antidepressants reported by means of a medicine reporting system

Objective To assess experiences related to antidepressant use reported to an internet-based medicine reporting system and to compare the nature of the side effects reported by patients with those reported by health care professionals (HCPs). Methods All reports submitted from May 2004 to May 2005 to an internet-based medicine reporting system in The Netherlands related to the use of antidepressants were analysed. Spontaneous reports of adverse drug reactions on antidepressants from HCPs received by The Netherlands Pharmacovigilance Centre Lareb from May 2004 to May 2005 were included for comparison. Results Of the 2232 individuals who submitted a report to the internet-based medicine reporting system, 258 submitted a report on antidepressants. Of these, 92 individuals (36%) reported on effectiveness, 40 (16%) of whom reported on ineffectiveness, and 217 (84%) submitted a report on side effects, with 202 (78%) reporting a total of 630 side effects that were experienced as negative. Fourteen individuals (5%) reported a practical issue and four (2%) reported a reimbursement issue. Of all 630 side effects reported, 48% resulted in the patient discontinuing the antidepressant therapy; of these 29% did not inform their HCP. Of all the side effects reported, 52% were perceived as “very negative”. In comparison to the side effects reported by HCPs, patients more often reported apathy, excessive sweating, ineffectiveness, somnolence, insomnia, sexual problems and weight increase. Conclusion Patients report the ineffectiveness and side effects of antidepressant therapy as negative and leading to discontinuation of the therapy. Patients and HCPs differ in the nature of the reported side effects. Patient experiences should be included in the evaluation of antidepressant treatment in clinical practice.     

How Much Binding Affinity Can be Gained by Filling a Cavity?

Binding affinity optimization is critical during drug development. Here, we evaluate the thermodynamic consequences of filling a binding cavity with functionalities of increasing van der Waals radii (–H, –F, –Cl, and CH₃) that improve the geometric fit without participating in hydrogen bonding or other specific interactions. We observe a binding affinity increase of two orders of magnitude. There appears to be three phases in the process. The first phase is associated with the formation of stable van der Waals interactions. This phase is characterized by a gain in binding enthalpy and a loss in binding entropy, attributed to a loss of conformational degrees of freedom. For the specific case presented in this article, the enthalpy gain amounts to − 1.5 kcal/mol while the entropic losses amount to +0.9 kcal/mol resulting in a net 3.5-fold affinity gain. The second phase is characterized by simultaneous enthalpic and entropic gains. This phase improves the binding affinity 25-fold. The third phase represents the collapse of the trend and is triggered by the introduction of chemical functionalities larger than the binding cavity itself [CH(CH₃)₂]. It is characterized by large enthalpy and affinity losses. The thermodynamic signatures associated with each phase provide guidelines for lead optimization.     

Does unintentional ingestion of ethylene glycol represent a serious risk?

The objective of this study was to evaluate data retrospectively on accidental ingestion of ethylene glycol (EG), based on calls to the Czech Toxicological Information Centre and from toxicological laboratories, in the years 2000-2004. All patients who ingested a known amount of EG and/or subjects with measured serum EG levels were included. A variety of clinical and laboratory parameters was collected. The medical records of 86 subjects, who had ingested from one to three swallows of EG, were analysed. The following findings emerged-metabolic acidosis (41%), vomiting (36%), nephrotoxicity (10%), and CNS depression (9%). In 15 children, the time interval between ingestion and hospitalisation was 1 hour or less. Ethanol was given to 12 children (four as first aid), and none developed hypoglycaemia. Of the 71 adults, 93% were treated with ethanol (19 as first aid). No side effects were documented. Seventeen patients received haemodialysis (HD). Two patients recovered without HD; their EG levels were higher than in the HD-treated patients. Unintentional EG ingestion usually involves ingestion of a small amount of EG, and was connected with mild signs of intoxication. Early therapy with ethanol alone appears sufficient in such cases, and represents no risk of adverse effects.     

How Well Can a T-Cell Epitope Replace Its Parent Carrier Protein? A Dose-Response Study

Purpose. This work examines the effectiveness of synthetic peptide immunogens derived from immunodominant T-cell epitopes as replacements for their intact parent protein in vaccines. Methods. Fluorescein was conjugated to hen egg lysozyme (FL-HEL, positive control) and three synthetic peptide immunogens: (a) murine B10.A (H-2^a) immunodominant T-cell epitope of HEL [FL-(T-cell epitope)]; (b) multiple antigenic peptide (MAP) multimer of this epitope {[FL-(T epitope)]_n-MAP, n = 2-4}; and (c) negative control MAP with T-cell epitope residues replaced with glycine [(FL-Gly₁₈)₄-MAP]. The dose response of each immunogen was examined over a 300-fold range in B10.A mice. The immune response was monitored using antifluorescein ELISA assays. Results. FL-(T epitope)'s immune response correlated positively with dose, with maximum response comparable to that of [FL-(T epitope)]_n-MAP, or FL-HEL. This trend was consistent across 1°, 2°, and 3° responses, although interanimal variability was higher in the latter two because of an all-or-none response in mice immunized with this peptide. [FL-(T epitope)]_n-MAP's immune response was consistently high and nearly dose independent, a trend observed across 1°, 2°, and 3° responses. FL-HEL's immune response correlated negatively to dose in the 1° response but was nearly dose independent in the 2° and 3° responses. The magnitude of these latter responses was comparable to that observed for [FL-(T epitope)]_n-MAP. (FL-Gly₁₈)₄-MAP did not elicit an immune response except at the highest dose. This trend was consistent across 1°, 2°, and 3° responses. Conclusions. The monomeric epitope was 300-fold less potent than its parent carrier protein, but increasing immunogen valency using MAP technology compensated totally for reduced potency. (FL-Gly₁₈)₄-MAP's lack of response at all but the highest dose strongly suggests that a specific immunodominant T-cell epitope sequence for HEL is necessary for successful peptide mimicry of HEL. This work also demonstrates the importance of quality assessment of commercial MAP core resins.     

Does post-translational modification influence chaperone-like activity of alpha-crystallin? I. Study on phosphorylation

It is difficult to isolate derivatives of alpha-crystallin with only one type of post-translational modification, because this protein is subjected to several different types of modification. In the present study using bovine lens proteins, we isolated mono-phosphorylated alphaB-crystallin with no other post-translational modifications. Using this material, we demonstrated that mono-phosphorylation reduced the activity of alphaB-crystallin by approximately 30%. Our results confirmed that investigation of the correlation between chaperone-like activities of alpha-crystallin and post-translational modification is important to understand the mechanism of cataract formation.     

Mechanisms of Change in Adolescent Substance Use Treatment: How Does Treatment Work?

ABSTRACT. Background: Adolescent substance use treatment outcome research generally shows small to moderate effects in reducing substance use, with no specific “brand” of treatment emerging as clearly superior to any other, and treatment gains that fade over time. The relatively weak and temporary effects of treatment call for improving the potency and durability of intervention effects. In response to this call, this critical narrative review summarizes research on mechanisms of change for both adults and adolescents in substance use treatment, with a particular focus on reviewing what is known regarding “how” adolescent substance use treatment works. Methods: A comprehensive review of the adolescent (aged 11–18) substance use treatment literature was conducted to identify empirical studies that examined mediators of intervention effects. Relevant databases (e.g., PsychINFO, MEDLINE) were searched using key words (e.g., “mediator”), and relevant articles from reference sections of identified studies and review papers were considered. Results: Studies of mechanisms of psychotherapy change are rare in the adult, and particularly adolescent, substance use treatment outcome literature. The 4 adolescent studies that examined substance use treatment mechanisms found that positive social support, motivation to abstain, and positive parenting behaviors mediated treatment effects. To date, research has not supported therapy-specific mechanisms of change, finding instead that “common” processes of change largely account for improvements in outcome across distinct “brands” of treatment. Conclusions: The lack of empirical support for treatment-specific mechanisms of change may be due to the need for greater precision in defining and measuring treatment-specific causal chains. Future directions include neuroscience approaches to examining changes in brain functioning that are associated with treatment response and recovery and examining mechanisms in adaptive treatment designs, which can accommodate individual differences in targets for intervention and response to treatment.     

Does a childhood history of externalizing problems predict smoking during pregnancy?

While the demographic characteristics of women who smoke during pregnancy are well established, less is known about psychiatric characteristics that may differentiate among persistent smokers, spontaneous quitters, and nonsmokers. The purpose of the present study was to test the hypothesis that a history of externalizing problems is related to persistent smoking during pregnancy. Participants included 93 pregnant women (mean age=28 years; 89% non-Hispanic White; 46% persistent smokers; and 16% spontaneous quitters). Externalizing problems, as evidenced by conduct disorder (CD) and attention deficit hyperactivity disorder (ADHD), were assessed using diagnostic interviews. History of CD and ADHD varied by smoking status, with persistent smokers most likely to have a history of both disorders and exhibiting the highest levels of symptomatology. In multivariate analyses, a history of CD, but not ADHD symptoms, distinguished women who persisted in smoking during pregnancy from spontaneous quitters. Results suggest that a childhood history of conduct problems is a risk factor for maternal smoking during pregnancy and that psychiatric history is important to consider in developing more targeted cessation interventions.     

Does risperidone have a place in the treatment of nonschizophrenic patients?

There is a now a substantial body of evidence that suggests the new antipsychotic agent, risperidone, may be safe and effective for treating psychotic, affective or behavioural symptoms associated with various disorders other than schizophrenia, schizophreniform disorder or schizo-affective disorder. These conditions include bipolar disorder, obsessive-compulsive disorder, Tourette's syndrome, dementia, Lewy body disease, mental retardation, Parkinson's disease, idiopathic segmental dystonia and organic catatonia. Although much of the data is anecdotal or in the form of open studies, there is now emerging a small number of well controlled investigations supporting efficacy for mania, dementia, behavioural disturbance in mental retardation and conduct disorder. Conventional antipsychotics have long been used, either in a primary capacity or as an adjunct to treat these disorders; however, they have limited benefit, pose significant risks of extrapyramidal side-effects, and may cause the potentially life-threatening neuroleptic malignant syndrome. In contrast, risperidone at the recommended low doses may be efficacious and pose reduced risk of motor side-effects. This article reviews the evidence that risperidone may be an effective new treatment for disorders other than schizophrenia.     

Does transbilayer diffusion have a role in membrane transport of drugs?

The existing consensus on coexistence of transbilayer diffusion and carrier-mediated transport as two main mechanisms for drugs crossing biological membranes was recently challenged by a systems biology group. Their transporters-only hypothesis is examined in this article using published experimental evidence. The main focus is on the key claim of their hypothesis, stating that 'the drug molecules cross pure phospholipid bilayers through transient pores that cannot form in the bilayers of cell membranes, and thus transbilayer drug transport does not exist in cells'. The analysis shows that the prior consensus remains a valid scientific view of the membrane transport of drugs.     

Can the Cellular Internalization of Cargo Proteins Be Enhanced by Fusing a Tat Peptide in the Center of Proteins? A Fluorescence Study

The aim of this study was to investigate whether the cellular uptake of cargo proteins can be enhanced by fusing a Tat peptide in the center of proteins; glutathione-S-transferase (GST)-Tat-green fluorescent protein (GFP) and GST-GFP-Tat proteins were first constructed and expressed. The cellular internalization of both proteins was then evaluated and compared in HeLa cells using fluorescent microscopy and flow cytometry, as well as the transdermal delivery in human skin using confocal microscopy. Results from in vitro cell experiments showed that GST-Tat-GFP protein efficiently internalized into HeLa cells when a Tat peptide was fused in the center of proteins, whereas its efficiency is lower than that of GST-GFP-Tat protein with a Tat peptide terminal fused. Ex vivo transdermal delivery data also demonstrated that the lower efficiency of GST-Tat-GFP penetrating through human stratum corneum layer when compared with GST-GFP-Tat. Furthermore, both GST-GFP-Tat and GST-Tat-GFP presented a various degree of a mixture of cytoplasmic diffuse staining and punctate surface staining, and the pattern of distribution varied considerably in HeLa cell experiments depending on the concentration of protein used. Therefore, an improved mechanism for Tat-conjugated proteins was proposed, in which Tat-conjugated proteins were first associated with cell membrane, then accumulated on the cell surface, and finally internalized into cells by pore formation mechanism.