Comprehensive safety assessment of serendipity berry sweet protein produced from Komagataella phaffii.

Serendipity berry plant (Dioscoreophyllum cumminsii (Stapf) Diels) is the source of a naturally sweet protein referred to as monellin. The safety of serendipity berry sweet protein (SBSP) containing single polypeptide monellin (MON) expressed in Komagataella phaffii (formerly Pichia pastoris) and produced via precision fermentation was examined comprehensively through assessments of in vitro and in silico protein digestion, in silico allergenicity, in vitro genotoxicity (reverse mutation and mammalian micronucleus assays), and 14-day and 90-day oral (dietary) toxicity studies in rats. There was no indication of allergenicity for SBSP in the in silico analyses. Results from both in vitro and in silico protein digestibility assessments indicated that SBSP is digested upon ingestion and would therefore be unlikely to pose a toxigenic or allergenic risk to consumers. SBSP was non-genotoxic in in vitro assays and showed no adverse effects in the 14-day or 90-day toxicity studies up to the highest dose tested. The 90-day toxicity study supports a NOAEL for SBSP of 1954 mg/kg bw/day, which corresponds to a NOAEL for MON of 408 mg/kg bw/day.

A toxicological assessment of spermidine trihydrochloride produced using an engineered strain of Saccharomyces cerevisiae.

Spermidine is a polyamine consumed in the diet, endogenously biosynthesized in most cells, and produced by the intestinal microbiome. A variety of foods contribute to intake of spermidine along with other polyamines. Spermidine trihydrochloride (spermidine-3HCl) of high purity can be produced using an engineered strain of Saccharomyces cerevisiae. Spermidine has a demonstrated history of safe use in the diet; however, limited information is available in the public literature to assess the potential toxicity of spermidine-3HCl. To support a safety assessment for this spermidine-3HCl as a dietary source of spermidine, authoritative guideline and good laboratory practice (GLP) compliant in vitro genotoxicity assays (bacterial reverse mutation and mammalian micronucleus assays) and a 90-day oral (dietary) toxicity study in rats were conducted with spermidine-3HCl. Spermidine-3HCl was non-genotoxic in the in vitro assays, and no adverse effects were reported in the 90-day oral toxicity study up to the highest dose tested, 12500 ppm, equivalent to 728 mg/kg bw/day for males and 829 mg/kg bw/day for females. The subchronic no observed adverse effect level (NOAEL) is 728 mg/kg bw/day.

Lack of genotoxicity potential and safety assessment of 4-(2-hydroxyethyl) morpholine present as an impurity in pharmaceuticals and nutritional supplements.

4-(2-Hydroxyethyl) morpholine (HEM) is widely used as a building block of macromolecules in the manufacture of pharmaceuticals and dietary supplements and could remain as an impurity in the finished products. An evaluation of HEM was conducted to identify endpoints that could be used to determine the point-of-departure (POD) for use in assessing the potential risk from exposure to HEM. No oral repeated dose toxicological studies of appropriate duration were found for HEM. Therefore, suitable analogue(s) were identified. Although oral repeated dose studies were available for the analogues, the studies were not of sufficient duration for use in the assignment of a POD for risk evaluation. Accordingly, the Threshold of Toxicological Concern (TTC) approach, which proposes that a de minimis value can be derived to qualitatively assess risk, was considered for HEM. To determine the appropriate TTC approach (genotoxic or non-genotoxic), the genotoxicity of HEM and its analogues were evaluated. The weight of the evidence indicated that HEM, and the appropriate analogues, are not genotoxic. Considering the chemical structure of HEM, the non-genotoxic Cramer class III TTC value of 1.5 µg/kg bw/day was determined to be appropriate for use in safety assessment of HEM as an impurity in products intended for human consumption.

A new approach methodology using kinetically-derived maximum dose levels in risk assessment - A case study with afidopyropen.

We present a case study for afidopyropen (AF; insecticide) to characterize chronic dietary human health risk using a Risk 21-based approach. Our objective is to use a well-tested pesticidal active ingredient (AF) to show how a new approach methodology (NAM), using the kinetically-derived maximum dose (KMD) and with far less animal testing, can reliably identify a health-protective point of departure (PoD) for chronic dietary human health risk assessments (HHRA). Chronic dietary HHRA involves evaluation of both hazard and exposure information to characterize risk. Although both are important, emphasis has been placed on a checklist of required toxicological studies for hazard characterization, with human exposure information only considered after evaluation of hazard data. Most required studies are not used to define the human endpoint for HHRA. The information presented demonstrates a NAM that uses the KMD determined by saturation of a metabolic pathway, which can be used as an alternative POD. In these cases, the full toxicological database may not need to be generated. Demonstration that the compound is not genotoxic and that the KMD is protective of adverse effects in 90-day oral rat and reproductive/developmental studies is sufficient to support the use of the KMD as an alternative POD.

Current testing programs for pesticides adequately capture endocrine activity and adversity for protection of vertebrate wildlife.

The toxicity and ecotoxicity of pesticide active ingredients are evaluated by a number of standardized test methods using vertebrate animals. These standard test methods are required under various regulatory programs for the registration of pesticides. Over the past two decades, additional test methods have been developed with endpoints that are responsive to endocrine activity and subsequent adverse effects. This article examines the available test methods and their endpoints that are relevant to an assessment of endocrine-disrupting properties of pesticides. Furthermore, the article highlights how weight-of-evidence approaches should be applied to determine whether an adverse response in (eco)toxicity tests is caused by an endocrine mechanism of action. The large number of endpoints in the current testing paradigms for pesticides make it unlikely that endocrine activity and adversity is being overlooked. Integr Environ Assess Manag 2023;19:1089-1109. © 2023 Bayer CropScience and The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Canine and feline P-glycoprotein deficiency: What we know and where we need to go.

In 2001 the molecular genetic basis of so-called "ivermectin sensitivity" in herding breed dogs was determined to be a P-glycoprotein deficiency caused by a genetic variant of the MDR1 (ABCB1) gene often called "the MDR1 mutation." We have learned a great deal about P-glycoprotein's role in drug disposition since that discovery, namely that P-glycoprotein transports many more drugs than just macrocyclic lactones that P-glycoprotein mediated drug transport is present in more places than just the blood brain barrier, that some cats have a genetic variant of MDR1 that results in P-glycoprotein deficiency, that P-glycoprotein dysfunction can occur as a result of drug-drug interactions in any dog or cat, and that the concept of P-glycoprotein "inhibitors" versus P-glycoprotein substrates is somewhat arbitrary and artificial. This paper will review these discoveries and discuss how they impact drug selection and dosing in dogs and cats with genetically mediated P-glycoprotein deficiency or P-glycoprotein dysfunction resulting from drug-drug interactions.

Using existing knowledge for the risk evaluation of crop protection products in order to guide exposure driven data generation strategies and minimise unnecessary animal testing.

Traditionally, human health risk assessment focuses on defining the hazard through mammalian toxicity studies followed by exposure estimation. We have explored ways of predicting exposure based primarily on the use scenario and comparing the exposure to reference dose values derived by various regulatory agencies (US EPA, JMPR, and EU Commission) in order to identify mammalian toxicity studies that are relevant to human health risk assessment. Human dietary exposure was based on existing residue data for substances with comparable use on the same or similar crops. Human occupational exposures were based on the use scenarios and application methods. To provide a point of comparison for the exposure predictions, data were collated for acute, chronic and occupational reference dose values derived by various regulatory agencies (US EPA, JMPR, and EU Commission). The exposure predictions and range of hazard endpoints were compared using the ILSI HESI Risk21 risk matrix plots in order to visualise and contextualise the level of potential concern for the exposure prediction. In addition, an approach is proposed to categorise the likelihood of acceptability of risk based on where the exposure sits relative to the distribution of reference dose values. The approaches proposed in this study allow for exposure prediction based on the Good Agricultural Practice (GAP) in conjunction with the use of existing hazard data for crop protection products in order to make an initial determination on acceptability of risk and to identify key studies that are required for human health risk assessment and also opportunities for study waivers.

Afidopyropen: Challenges and impact of a toxicokinetic study designed to identify a point of inflection from dose-proportionality.

Afidopyropen is an insecticide that acts as a transient receptor potential vanilloid subtype (TRPV) channel modulator in chordotonal organs of target insects and has been assessed for a wide range of toxicity endpoints including chronic toxicity and carcinogenicity in rats and mice. The current study evaluates the toxicokinetic properties of afidopyropen and its plasma metabolites in rats at dose levels where the pharmacokinetics (PK) are linear and nonlinear in an attempt to identify a point of inflection. Based on the results of this study and depending on the analysis method used, the kinetically derived maximum dose (KMD) is estimated to be between 2.5 and 12.5 mg/kg bw/d with linearity observed at doses below 2.5 mg/kg bw/d. A defined point of inflection could not be determined. These data demonstrate that consideration of PK is critical for improving the dose-selection in toxicity studies as well as to enhance human relevance of the interpretation of animal toxicity studies. The study also demonstrates the technical difficulty in obtaining a defined point of inflection from in vivo PK data.

Regulatory developments and their impacts to the nano-industry: A case study for nano-additives in 3D printing.

Nanotechnology has increasing applications in numerous markets, particularly in additive processing (3D printing) and manufacturing, which is important for consumer products, medical devices, construction, and general research and development across many other industries. Nanomaterials are desirable in many products due to their unique properties, but those same properties have made evaluating the risk and regulation of these materials challenging. For risk-based regulations, new applications and nanomaterials should be assessed for both human and environmental hazards and exposure to ensure protection. In general, many risk assessments to date have focused on the non-nano versions of chemicals. The lack of guidance on assessing the hazard and exposure of nanomaterials in 3D printing is apparent, and these areas of assessment are actively being evaluated. Industry in most cases will now need to provide specific additional information for assessing the risk of nanomaterials in 3D printing. This review paper focuses on the use of nanomaterials in 3D printing for industrial and manufacturing applications, summarizes the current literature on human health and safety related to 3D printing and inhalation exposure, and the regulations relating to 3D printing in the U.S., Canada, and Europe for this industry.

Critical Review of Read-Across Potential in Testing for Endocrine-Related Effects in Vertebrate Ecological Receptors.

Recent regulatory testing programs have been designed to evaluate whether a chemical has the potential to interact with the endocrine system and could cause adverse effects. Some endocrine pathways are highly conserved among vertebrates, providing a potential to extrapolate data generated for one vertebrate taxonomic group to others (i.e., biological read-across). To assess the potential for biological read-across, we reviewed tools and approaches that support species extrapolation for fish, amphibians, birds, and reptiles. For each of the estrogen, androgen, thyroid, and steroidogenesis (EATS) pathways, we considered the pathway conservation across species and the responses of endocrine-sensitive endpoints. The available data show a high degree of confidence in the conservation of the hypothalamus-pituitary-gonadal axis between fish and mammals and the hypothalamus-pituitary-thyroid axis between amphibians and mammals. Comparatively, there is less empirical evidence for the conservation of other EATS pathways between other taxonomic groups, but this may be due to limited data. Although more information on sensitive pathways and endpoints would be useful, current developments in the use of molecular target sequencing similarity tools and thoughtful application of the adverse outcome pathway concept show promise for further advancement of read-across approaches for testing EATS pathways in vertebrate ecological receptors. Environ Toxicol Chem 2020;39:739-753. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

'A frightening experience': detainees' and carers' experiences of being detained under Section 136 of the Mental Health Act.

AIMS: This research reports on the views of detainees and their carers of their experiences of being detained under Section 136 (S136) of the Mental Health Act 1983. Individual interviews were conducted with 18 detainees and six carers. METHOD: A semi-structured questionnaire was administered face-to-face to gather qualitative data, which was analysed using a grounded theory approach. RESULTS: The results indicated a general dissatisfaction with the quality of care and treatment from both police and professionals. Though several detainees recognized the need for police to be involved, most felt they lacked the skills needed to meet the needs of mentally ill people. Nearly all participants felt that the police station was an inappropriate setting for further assessment, and found their experiences in police cells distressing, making them feel like criminals. CONCLUSIONS: Detainees and carers would like to see the provision of a place of safety other than emergency departments or police stations, and this study reinforces the Mental Health Code of Practice 2008 which states that police stations should only be used on an exceptional basis.

Overcoming the barriers to chlamydia screening in general practice--a qualitative study.

BACKGROUND: There is low uptake of chlamydia screening in general practices registered with the English National Chlamydia Screening Programme (NCSP). Aims. To explore staff's attitudes and behaviour around chlamydia screening and how screening could be optimized in general practice. METHODS: A qualitative study with focus groups and interviews, in general practices in seven NCSP areas. Twenty-five focus groups and 12 interviews undertaken with a purposively selected diverse group of high and low chlamydia-screening practices in 2006-08. Data were collected and analysed using a framework analytical approach. RESULTS: Higher screening practices had more staff with greater belief in patient and population benefits of screening and, as screening was a subjective norm, it was part of every day practice. Many staff in the majority of other practices were uncomfortable raising chlamydia opportunistically and time pressures meant that any extra public health issues covered within a consultation were determined by Quality Outcomes Framework (QOF) targets. All practices would value more training and feedback about their screening rates and results. Practices suggested that use of computer prompts, simplified request forms and more accessible kits could increase screening. CONCLUSION: Practice staff need more evidence of the value of opportunistic chlamydia screening in men and women; staff development to reduce the barriers to broaching sexual health; simpler request forms and easily accessible kits to increase their ability to offer it within the time pressures of general practice. Increased awareness of chlamydia could be attained through practice meetings, computer templates and reminders, targets and incentives or QOF points with feedback.

Promoting chlamydia screening with posters and leaflets in general practice--a qualitative study.

BACKGROUND: General practice staff are reluctant to discuss sexual health opportunistically in all consultations. Health promotion materials may help alleviate this barrier. Chlamydia screening promotion posters and leaflets, produced by the English National Chlamydia Screening Programme (NCSP), have been available to general practices, through local chlamydia screening offices, since its launch. In this study we explored the attitudes of general practice staff to these screening promotional materials, how they used them, and explored other promotional strategies to encourage chlamydia screening. METHODS: Twenty-five general practices with a range of screening rates, were purposively selected from six NCSP areas in England. In focus groups doctors, nurses, administrative staff and receptionists were encouraged to discuss candidly their experiences about their use and opinions of posters, leaflets and advertising to promote chlamydia screening. Researchers observed whether posters and leaflets were on display in reception and/or waiting areas. Data were collected and analysed concurrently using a stepwise framework analytical approach. RESULTS: Although two-thirds of screening practices reported that they displayed posters and leaflets, they were not prominently displayed in most practices. Only a minority of practices reported actively using screening promotional materials on an ongoing basis. Most staff in all practices were not following up the advertising in posters and leaflets by routinely offering opportunistic screening to their target population. Some staff in many practices thought posters and leaflets would cause offence or embarrassment to their patients. Distribution of chlamydia leaflets by receptionists was thought to be inappropriate by some practices, as they thought patients would be offended when being offered a leaflet in a public area. Practice staff suggested the development of pocket-sized leaflets. CONCLUSION: The NCSP should consider developing a range of more discrete but eye catching posters and small leaflets specifically to promote chlamydia screening in different scenarios within general practice; coordinators should audit their use. Practice staff need to discuss, with their screening co-ordinator, how different practice staff can promote chlamydia screening most effectively using the NCSP promotional materials, and change them regularly so that they do not loose their impact. Education to change all practice staff's attitudes towards sexual health is needed to reduce their worries about displaying the chlamydia materials, and how they may follow up the advertising up with a verbal offer of screening opportunistically to 15-24 year olds whenever they visit the practice.

Poetry in general practice education: perceptions of learners.

BACKGROUND: There has been little research into poetry-based medical education. Few studies consider learners' perceptions in depth. OBJECTIVE: To explore general practice registrars' (GPRs) perceptions of two poetry-based sessions. METHODS: GPRs in one general practice vocational training scheme experienced two poetry sessions. In one, the facilitator selected poems; in the other, poems were chosen by registrars. Poems were read and discussed, with emphasis on personal response. Data were obtained through in-depth semi-structured interviews with six registrars. Interviews were audiotaped, transcribed and analysed using interpretative phenomenological analysis. Identification of individual ideas and shared themes enabled exploration of the registrars' experiences. RESULTS: Registrars described how poetry helped them explore emotional territory. They recognized a broadening of education, describing how poems helped them consider different points of view, increasing their understanding of others. Vicarious experience, development of empathy and self-discovery were also reported. Participants speculated on how this might impact on patient care and professional practice. Facilitator-selected poems provided variety and ambiguity, provoking discussions with clinical relevance. Learner-selected poems enabled involvement, self-revelation and understanding of peers and developed emotional expression. CONCLUSIONS: These registrars reported difficulties expressing feelings in the culture of science-based medical training. Poetry sessions may provide an environment for emotional exploration, which could broaden understanding of self and others. Poetry-based education may develop emotional competence. The participants recognized development of key skills including close reading, attentive listening and interpretation of meaning. These skills may help doctors to understand individual patient's unique experience of illness, encouraging personalized care that respects patients' perspectives.

'Look after your legs': patients' experience of an assessment clinic.

Venous leg ulcers are a chronic, recurring condition and a major cause of morbidity, especially in older people. Two inner city district nursing teams initiated a weekly, nurse-led, leg ulcer assessment clinic, and a monthly support group for patients with healed or almost healed leg ulcers. The group supports and empowers patients with self-care 'look after your legs' health promotion messages. The support group uses a similar model to the expert patient programme (Department for Health, 2001). Patients in the group coined the unique name 'patient ambassador'. Patient ambassadors, with nursing support, are used to promote self care and lifestyle changes to members of the group to prevent recurrence of leg ulceration. The aim of the research reported here was to evaluate teh 'Look after Your Legs'(LAYL) service and the benefit to others of the patient ambassador role.

Helicobacter pylori test & treat strategy for dyspepsia: a qualitative study exploring the barriers and how to overcome them.

BACKGROUND: The National Institute of Clinical Excellence (NICE) has issued guidance that recommends that all dyspeptic patients without alarm symptoms, irrespective of age, that relapse after one month's proton pump inhibitor, should be offered test and treat for Helicobacter pylori. OBJECTIVE: To explore the views of primary care about introducing the helicobacter test and treat NICE dyspepsia guidance. METHODS: In 15 urban and rural general practices in Central England, primary health care staff involved in the management of dyspepsia took part in qualitative focus groups to discuss the draft NICE guidance and how it might be implemented. RESULTS: Practices expressed concern that test and treat may not be cost-effective in younger patients and that they may miss malignancy, in older patients and in patients who relapse after triple treatment, without a further helicobacter test or endoscopy. The greatest practical barriers to test and treat were the considerable impact on nurses' and doctors' time to explain, undertake tests and report results to patients, and practice budgets from urea breath test and triple treatment costs. Staff preferred stool tests to breath tests, as they impacted less on practice budget and time. GPs did not favour prescribing the three components of the triple treatment separately. CONCLUSIONS: GPs will need reassurance that test and treat will not lead to missed malignancies. The financial costs and staffing implications of NICE dyspepsia guidance will need to be discussed locally by Primary Care Trusts, microbiology laboratories, gastroenterologists and pharmacy advisors and implemented with local guidance, increased communication and education.

Is the Palliative Care Outcome Scale useful to staff in a day hospice unit?

AIM: this study was undertaken to elicit a day hospice team's experience of using the Palliative Care Outcome Scale (POS), with the intention of determining its usefulness to staff. An adapted version of POS was used for the purpose of the study. SAMPLE: a multiprofessional team of eight day hospice staff took part in the study. All had used POS on a weekly basis for a minimum period of 3 months. DESIGN: a focus group was conducted at the hospice. It was tape-recorded and transcribed verbatim. The data were analysed using an interpretive phenomenological approach. RESULTS: staff felt POS had limited value as an outcome measure for use in this day hospice. Other simultaneous organizational changes at the hospice may also have influenced staff negatively. Further research is needed to explore similar implementation issues in more depth, as practice developments may be abandoned for potentially the wrong reasons.

The prevalence of dysfunctional breathing in adults in the community with and without asthma.

Functional breathing problems, including symptomatic hyperventilation, may impair quality of life. Symptoms associated with functional breathing disorders have been reported as being common in secondary care settings, and can affect 29% of adults with current asthma in the community. The prevalence of dysfunctional breathing in the general adult population is unknown. The Nijmegen Questionnaire has been reported to have useful sensitivity and specificity for diagnosing dysfunctional breathing. A cross-sectional postal survey of adults without current asthma was undertaken in a single UK general practice. The results were analysed in conjunction with a previously described survey of adults with current asthma from the same population. The questionnaire was posted to a random sample of 300 people aged 16-65 without current asthma, and 69% were returned. 8% (95% confidence intervals 4-12%) had positive screening scores. Positive screening scores were more common in women (14%, 7-20%) than men (2%, 0-5%, p=0.003). Comparison with the previous survey showed that the prevalence of positive screening scores was higher in those with current asthma than those without (29% vs. 8%, p<0.001). Dysfunctional breathing may affect up to one in 10 people, and is more common in women and in people with asthma.

Patients' views of using an outcome measure in palliative day care: a focus group study.

Outcome measures are being increasingly used in palliative care. Palliative day care is significantly different to other specialist palliative care settings, as patients may attend for much longer periods of time. The aim of this study was to elicit patients' views on using an adapted version of the Palliative Care Outcome Scale (POS) and to determine its value in identifying their individual needs. This qualitative study involved a focus group of nine patients who had used POS weekly for a minimum of 3 months. The focus group discussion was tape recorded and transcribed verbatim. Data were analysed using an interpretive phenomenological approach. Participants found POS to be a useful communication tool, which helped them to identify their individual needs.