The influence of antibodies to TNF-α and IL-1β on haemodynamic responses to the cytokines,and to lipopolysaccharide,in conscious rats

1. Male, Long Evans rats (350–450 g) were anaesthetized and had pulsed Doppler probes and intravascular catheters implanted to allow monitoring of regional (renal, mesenteric and hindquarters) haemodynamics in the conscious state. Our main objectives were to:- assess the effects of administering human recombinant tumour necrosis factor (TNF)-α and human recombinant interleukin-1 (IL-1)β, alone and together; determine the influence of pretreatment with a mixture of antibodies to TNF-α and IL-1β on responses to co-administration of the cytokines; ascertain if pretreatment with a mixture of the antibodies to TNF-α and IL-1β had any influence on the responses to lipopolysaccharide (LPS).
2. TNF-α (10, 100 and 250 μg kg⁻¹, in separate groups, n=3, 9 and 8, respectively) caused tachycardia (maximum Δ, +101±9 beats min⁻¹) and modest hypotension (maximum Δ, −10±2 mmHg), accompanied by variable changes in renal and mesenteric vascular conductance, but clear increases in hindquarters vascular conductance; only the latter were dose-related (maximum Δ, +6±6, +27±9, and +61±12% at 10, 100 and 250 μg kg⁻¹, respectively).
3. IL-1β (1, 10, and 100 μg kg⁻¹ in separate groups, n=8, 8 and 9, respectively) evoked changes similar to those of TNF-α (maximum Δ heart rate, +69±15 beats min⁻¹; maximum Δ mean blood pressure, −14±2 mmHg; maximum Δ hindquarters vascular conductance, +49±17%), but with no clear dose-dependency.
4. TNF-α (250 μg kg⁻¹) and IL-1β (10 μg kg⁻¹) together caused tachycardia (maximum Δ, +76±15 beats min⁻¹) and hypotension (maximum Δ, −24±2 mmHg) accompanied by increases in renal, mesenteric and hindquarters vascular conductances (+52±6%, +23±8%, and +52±11%, respectively). Thereafter, blood pressure recovered, in association with marked reductions in mesenteric and hindquarters vascular conductances (maximum Δ, −50±3% and −58±3%, respectively). Although bolus injection of LPS (3.5 mg kg⁻¹) caused an initial hypotension (maximum Δ, −27±11 mmHg) similar to that seen with co-administration of the cytokines, it did not cause mesenteric or hindquarters vasodilatation, and there was only a slow onset renal vasodilatation. The recovery in blood pressure following LPS was less than after the cytokines, and in the former condition there was no mesenteric vasoconstriction. By 24 h after co-administration of TNF-α and IL-1β or after bolus injection of LPS, the secondary reduction in blood pressure was similar (−16±2 and −13±3 mmHg, respectively), but in the former group the tachycardia (+117±14 beats min⁻¹) and increase in hindquarters vascular conductance (+99±21%) were greater than after bolus injection of LPS (+54±16 beats min⁻¹ and +43±9%, respectively).
5. Pretreatment with antibodies to TNF-α and IL-1β (300 mg kg⁻¹) blocked the initial hypotensive and mesenteric and hindquarters vasodilator responses to co-administration of the cytokines subsequently. However, tachycardia and renal vasodilatation were still apparent. Premixing antibodies and cytokines before administration prevented most of the effects of the latter, but tachycardia was still present at 24 h.
6. Pretreatment with antibodies to TNF-α and IL-1β before infusion of LPS (150 μg kg⁻¹ h⁻¹ for 24 h) did not affect the initial fall in blood pressure, but suppressed the hindquarters vasodilatation and caused a slight improvement in the recovery of blood pressure. However, pretreatment with the antibodies had no effect on the subsequent cardiovascular sequelae of LPS infusion.
7. The results indicate that although co-administration of TNF-α and IL-1β can evoke cardiovascular responses which, in some respects, mimic those of LPS, and although antibodies to the cytokines can suppress most of the cardiovascular effects of the cytokines, the antibodies have little influence on the haemodynamic responses to LPS, possibly because, during infusion of LPS, the sites of production and local action of endogenous cytokines, are not accessible to exogenous antibodies.

     

Kava,the anxiolytic herb: back to basics to prevent liver injury?

The use of the anxiolytic herb kava has caused toxic liver injury in Western countries and economic problems in South Pacific Islands due to tthe regulatory ban on kava. This analysis shows poor quality of kava raw material as a cause for its toxicity and suggests preventative measures by going back to the traditional use of kava for the sake of the patients and the South Pacific economy.     

Binding ofβ-adrenoceptor blocking drugs to human serum albumin,to α1-acid glycoprotein and to human serum

Summary The binding of 8 -adrenergic blocking drugs to human serum albumin, to ₁-acid glycoprotein and to serum from normal volunteers and from patients with rheumatoid arthritis was studied. Protein binding was determined in vitro using equilibrium dialysis of labelled drug at 25° C. Oxprenolol and propranolol were highly bound to serum, alprenolol, pindolol and timolol to a lesser degree, and atenolol, metoprolol and sotalol were negligibly bound. For the five compounds which were appreciably bound, the mean binding was significantly higher in serum from patients with rheumatoid arthritis than in serum from normal volunteers. For those drugs, binding to ₁-acid glycoprotein was higher than to human serum albumin, and binding to a mixture of both proteins approached that to serum from healthy volunteers. For each of these drugs there was a strong correlation between the serum ₁-glycoprotein concentration and the percentage binding.     

Development, Validation, and Clinical Implementation of an Assay to Measure Total Antibody Response to Naglazyme® (Galsulfase)

Naglazyme® (galsulfase, rhASB) was developed as enzyme replacement therapy for mucopolysaccharidosis type VI. Naglazyme generated an IgG antibody response in most patients. To better characterize Naglazyme immunogenicity, a solution phase bridged immunoassay was developed to measure total antibody response regardless of isotype. Overnight incubation of serum dilutions with rhASB labeled with biotin and ruthenium-based tags allowed antibody–antigen complexes to form prior to capture on a streptavidin plate. Neat serum was tolerated in the assay, with a 1:10 screening dilution implemented for testing. At this dilution, the assay was sensitive to 75 ng/ml anti-rhASB. Titers were reported as the highest dilution factor with signal above a 95% confidence interval from naïve individual sera. Precise measurement of titers, within two consecutive dilution factors, was observed across analysts and days. Clinical samples showed similar positive/negative results between the IgG ELISA and the total antibody ECLA, although with an imperfect correlation. Improvements in assay performance and implementation strategy altered some positive clinical samples to negative and vice versa. Comparison of the titer readout for clinical samples with the screening signal illustrates a range of relationships for signal versus sample dilution factor, confirming that signal from a screening dilution cannot directly predict the reported titer.     

Deposition, imaging, and clearance: what remains to be done?

Deposition and clearance studies are used during product development and in fundamental research. These studies mostly involve radionuclide imaging, but pharmacokinetic methods are also used to assess the amount of drug absorbed through the lungs, which is closely related to lung deposition. Radionuclide imaging may be two-dimensional (gamma scintigraphy or planar imaging), or three-dimensional (single photon emission computed tomography and positron emission tomography). In October 2009, a group of scientists met at the "Thousand Years of Pharmaceutical Aerosols" conference in Reykjavik, Iceland, to discuss future research in key areas of pulmonary drug delivery. This article reports the session on "Deposition, imaging and clearance." The objective was partly to review our current understanding, but more importantly to assess "what remains to be done?" A need to standardize methodology and provide a regulatory framework by which data from radionuclide imaging methods could be compared between centers and used in the drug approval process was recognized. There is also a requirement for novel radiolabeling methods that are more representative of production processes for dry powder inhalers and pressurized metered dose inhalers. A need was identified for studies to aid our understanding of the relationship between clinical effects and regional deposition patterns of inhaled drugs. A robust methodology to assess clearance from small conducting airways should be developed, as a potential biomarker for therapies in cystic fibrosis and other diseases. The mechanisms by which inhaled nanoparticles are removed from the lungs, and the factors on which their removal depends, require further investigation. Last, and by no means least, we need a better understanding of patient-related factors, including how to reduce the variability in pulmonary drug delivery, in order to improve the precision of deposition and clearance measurements.     

Smoking Cessation Leads to Reduced Stress,but Why?

Recent longitudinal studies have demonstrated that smoking cessation leads to reduced feelings of stress. This finding is not predicted by either of the two main models for smoking behavior. The nicotine resource model (Warburton) states that nicotine is used to cope with external stressors, and predicts that smokers will suffer from increased stress when they quit smoking. The deprivation reversal model (Schachter), suggests that smoking reverses the deleterious effects of deprivation; cessation will then lead to a period of increased stress, followed by a return to baseline. Although the stress/cessation data agree with neither model, they are consistent with a third explanation, namely that smoking causes stress. This model states that acute nicotine deprivation (i.e., between cigarettes) leads to increased stress. Smokers then use cigarettes to reverse these withdrawal effects and “normalize” their mood. This model explains some paradoxical aspects of the smoking/mood relationship. First, why smokers are calmed by smoking, yet report high average levels of stress. Second, why stress levels become reduced after smoking cessation; this is because the former smoker no longer suffers from the adverse mood effects of acute nicotine depletion.     

RAMPs: 5 years on, where to now?

It is now approximately 5 years since the identification of the family of receptor activity modifying proteins (RAMPs). This finding revolutionized concepts of the pharmacology of G-protein-coupled receptors (GPCRs) and revealed that GPCR accessory proteins not only assist trafficking and folding but also define receptor type. Since the identification of RAMPs as modulators of the trafficking and properties of the calcitonin-receptor-like receptor, much work has focused on improving our understanding of the nature of RAMP-GPCR dimers, the extent to which they occur, and the consequence of this association. In this article, we review recent developments, including the identification of new receptor partners and novel roles for RAMPs.     

Every child to thrive, belong and achieve? Time to reflect and act in New Zealand

New Zealand continues to grapple with poor and inequitable child health and wellbeing outcomes. The associated high economic costs, the long-term impact on adult health and New Zealand's international children's rights obligations provide further grounds for action. Although there have been many different reports offering solutions and some key areas of progress, gains have been limited and there has not been sufficient clarity and agreement on wider actions. The environment is complex and solutions cross agency and disciplinary boundaries. This paper reviews the current situation and proposes a set of actions to improve child health and equity. These include a group of recommendations on high-level leadership and coordination, actions to address social conditions, and a range of specific health and wellbeing actions. Progress will require the will, commitment and courage of many to acknowledge the issues and find a way forward. Preventing suffering and ensuring the wellbeing of our youngest citizens during their formative years is an ethical issue for our nation, an issue of what we value as a society, and the best investment for a highly productive, innovative and resilient nation for the future.     

Community pharmacists’ attitudes,barriers, knowledge and counseling practice with regard to preconception,pregnancy and lactation

BackgroundAlthough preferably avoided, pregnant and lactating women often use medicines, including potentially inappropriate products. Pregnant women also commonly search for online information on medication. This underscores the need for appropriate counseling by healthcare professionals, including community pharmacists. However, little is known about pharmacists’ perceptions and barriers towards this role, nor about their knowledge and counseling practice.ObjectivesTo explore licensed pharmacists’ attitudes, barriers, knowledge and counseling practice regarding pharmaceutical care during preconception, pregnancy and lactation.MethodsAll pharmacists employed in 40 randomly selected pharmacies of a Belgian pharmacy chain (‘Surplus Network’) were asked to complete an online survey between September 2018–February 2019. The survey assessed pharmacists' attitudes and current practice regarding their role, as well as barriers and knowledge. Additionally, the 40 pharmacies were visited twice in December 2017 by simulated patients. The scenarios included a request for a pregnancy test and for an over-the-counter (OTC) product to treat pregnancy-related nausea.ResultsAll 63 invited pharmacists completed the survey. For all attitude-related statements, at least 80% agreed that pharmacists should take up the presented roles. However, for only 1 in 5 statements did at least 80% report currently taking up these roles for all or most patients. Most commonly reported barriers related to difficulties identifying the woman's status (71%) and lack of education (67%). The median score for the knowledge test was 23/45 (range: 6–36). The mystery shopping showed inadequate questioning and insufficient counseling. Only in 10% of cases was folic acid intake spontaneously discussed; in 39% of cases, incorrect advice about the dispensed OTC-product was given.ConclusionPharmacists acknowledged having an important role during preconception, pregnancy and lactation, but currently do not provide pharmaceutical care in this area to most patients. Educational programs are urgently needed to improve licensed pharmacists’ knowledge and counseling regarding this topic.     

Acute stroke in patients on new direct oral anticoagulants: how to manage,how to treat?

Introduction: For a long time, vitamin K antagonists (VKA) were the only available oral anticoagulants for clinical use. It is conceivable that the number of patients treated with novel direct oral anticoagulants (NOAC) will increase, due to the easy handling and the favorable risk–benefit profile compared with VKA. It is, therefore, expected that clinicians will be increasingly confronted with the question on how to treat acute ischemic stroke (AIS) if there is an indication for thrombolysis or how to manage intracranial bleedings.

Areas covered: In this review, we discuss controversies on thrombolysis in patients anticoagulated with NOAC, the dilemma of when to restart anticoagulation after AIS, and whether (and when) to re-institute oral anticoagulation after a brain hemorrhage. We provide suggestions for the management of these situations.

Expert opinion: Thrombolysis for patients with ischemic stroke who were given warfarin at subtherapeutic International normalized ratio values (≤ 1.7) may be considered according to guideline. Thrombolysis is contraindicated if intake of NOAC is reported in a patient, but no other information is available on-time of last intake of NOAC. Prothrombin complex concentrate have been proposed as a plausible, but unproven therapy to reverse the anticoagulant effects of NOACs.     

Initial opportunity to use marijuana and the transition to first use: United States, 1979–1994

A renewed American interest in marijuana has coincided with our research group's focus on the earliest stages of drug involvement. Here, we have studied the transition from an initial opportunity to try marijuana to the subsequent use of this drug. We analyzed self-report interview data gathered from nationally representative samples of the United States National Household Surveys on Drug Abuse, 1979–1994. The evidence indicates that the estimated prevalence of an opportunity to try marijuana has been rather stable for 15 years. However, there are recent increases in the probability of rapidly progressing from first marijuana opportunity to first marijuana use, among persons given an opportunity to use. In addition, the transition from first marijuana opportunity to eventual marijuana use seems to depend upon age at first opportunity. This epidemiological evidence on the transition from marijuana opportunity to marijuana use, the first to be published based on a nationally representative US sample, highlights directions for future research and a focus for prevention efforts.     

Plasma IL-2, NK, IFN-γ, and C3 in male workers exposed to traffic pollutants

The aim of the study is to evaluate if the occupational exposure to urban stressors could cause alterations in interleukin-2 (IL-2), NK, interferon-γ (IFN-γ) and C3 plasma levels in male traffic police officers compared to controls. After excluding the principal confounding factors, 108 traffic police officers were matched with 108 controls by age, working life, habitual consumption of alcohol and spirits. IL-2 mean levels were significantly higher in traffic police officers compared to controls (p=0.04). The distribution of IL-2 values in traffic police officers and in controls was significant (p=0.01). The distribution of NK value percentage in traffic police officers and in controls was significant (p=0.000). IFN-γ and C3 mean levels were not significant in traffic police officers compared to controls. Our results suggest that the occupational chronic exposure to low doses of urban stressors could affect NK and IL-2 plasma concentrations in traffic police officers of male sex.     

A Ligand-Based Approach to Understanding Selectivity of Nuclear Hormone Receptors PXR,CAR, FXR,LXRα, and LXRβ

Pharmaceutical Research - In recent years discussion of nuclear hormone receptors, transporters, and drug-metabolizing enzymes has begun to take place as our knowledge of the overlapping ligand...