Acceleration of infectious disease drug discovery and development using a humanized model of drug metabolism.

A key step in drug discovery, common to many disease areas, is preclinical demonstration of efficacy in a mouse model of disease. However, this demonstration and its translation to the clinic can be impeded by mouse-specific pathways of drug metabolism. Here, we show that a mouse line extensively humanized for the cytochrome P450 gene superfamily ("8HUM") can circumvent these problems. The pharmacokinetics, metabolite profiles, and magnitude of drug-drug interactions of a test set of approved medicines were in much closer alignment with clinical observations than in wild-type mice. Infection with Mycobacterium tuberculosis, Leishmania donovani, and Trypanosoma cruzi was well tolerated in 8HUM, permitting efficacy assessment. During such assessments, mouse-specific metabolic liabilities were bypassed while the impact of clinically relevant active metabolites and DDI on efficacy were well captured. Removal of species differences in metabolism by replacement of wild-type mice with 8HUM therefore reduces compound attrition while improving clinical translation, accelerating drug discovery.

Structure-Guided Design and Synthesis of a Pyridazinone Series of Trypanosoma cruzi Proteasome Inhibitors.

There is an urgent need for new treatments for Chagas disease, a parasitic infection which mostly impacts South and Central America. We previously reported on the discovery of GSK3494245/DDD01305143, a preclinical candidate for visceral leishmaniasis which acted through inhibition of the Leishmania proteasome. A related analogue, active against Trypanosoma cruzi, showed suboptimal efficacy in an animal model of Chagas disease, so alternative proteasome inhibitors were investigated. Screening a library of phenotypically active analogues against the T. cruzi proteasome identified an active, selective pyridazinone, the development of which is described herein. We obtained a cryo-EM co-structure of proteasome and a key inhibitor and used this to drive optimization of the compounds. Alongside this, optimization of the absorption, distribution, metabolism, and excretion (ADME) properties afforded a suitable compound for mouse efficacy studies. The outcome of these studies is discussed, alongside future plans to further understand the series and its potential to deliver a new treatment for Chagas disease.

Integrating Treatment for Maternal Depression and Young Children's Behavior Problems.

It is important to consider reciprocal associations between maternal and offspring mental health problems during early childhood. Existing interventions often focus narrowly on either adult or child mental health, missing the opportunity for holistic care. We describe the rationale and development of a pilot randomized clinical trial that explores their integration, combining an evidence-based parenting intervention with depression treatment to improve both maternal and child outcomes. Our approach is part of a growing field of two-generation interventions that offer a promising approach to enhance mental health support for caregivers and their young children.

A Caregiver-Child Intervention for Mitigating Toxic Stress ("The Resiliency Clinic"): A Pilot Study.

INTRODUCTION: Primary care-based interventions that promote nurturing caregiving relationships and early relational health may help mitigate toxic stress and promote resilience in children. This pilot study aims to: (1) describe a novel group-based, psychoeducational primary care intervention for children experiencing adverse childhood experiences (ACEs) ("The Resiliency Clinic"), (2) assess program feasibility and acceptability, and (3) explore effects on child/caregiver behavioral health. METHODS: Intervention design centered on promoting supportive caregiving, caregiver/child self-regulation and co-regulation and teaching evidence-based stress management tools. Program feasibility and acceptability were assessed through attendance data and caregiver focus groups. Behavioral health measures were obtained at baseline and 8-month follow-up. RESULTS: Of 101 eligible families, 38 (37.6%) enrolled and attended a median of 3.00 (mean = 2.95, sd = 1.75) out of 6 sessions. Caregivers reported high satisfaction and benefits including stress management tools and connection with staff and other parents. There were modest, statistically non-significant improvements in caregiver stress (d = 0.23) and child executive functioning (d = 0.27). DISCUSSION: In conclusion, a group intervention teaching supportive caregiving and stress mitigation is feasible and acceptable for many families in an urban federally qualified health center (FQHC) with a signal for modest improvements in behavioral health. Future program iterations will seek to address participation barriers and expand the intervention's capacity to promote early relational health.

Development of a label-free Raman imaging technique for differentiation of malaria parasite infected from non-infected tissue.

During malarial infection, the host uses the spleen to clear the malaria parasites, however, the parasites have evolved the ability to bind to endothelial receptors in blood vessels of tissues to avoid removal, known as sequestration, and this is largely responsible for the symptoms and severity of infection. So a technique which could non-invasively diagnose tissue burden could be utilised as an aid for localised malaria diagnosis within tissue. Raman spectroscopy is a label-free imaging technique and can provide unique and chemically specific Raman 'fingerprint' spectrum of biological samples such as tissue. Within this study, Raman imaging was used to observe the changes to the molecular composition of mice spleen tissue under malarial infection, compared with non-infected samples. From analysis of the Raman imaging data, both tissue types showed very similar spectral profiles, which highlighted that their biochemical compositions were closely linked. Principal component analysis showed very clear separation of the two sample groups, with an associated increase in concentration of heme-based Raman vibrations within the infected dataset. This was indicative of the presence of hemozoin, the malaria pigment, being detected within the infected spleen. Separation also showed that as the hemozoin content within the tissue increased, there was a corresponding change to hemoglobin and some lipid/nucleic acid vibrations. These results demonstrate that Raman spectroscopy can be used to easily discriminate the subtle changes in tissue burden upon malarial infection.

Marriage education for clinicians.

The field of marriage education has come to be dominated by nonprofessionals with no clinical training because clinicians interested in relationships typically provide marital therapy to couples in distress rather than marriage education to healthy couples. In this paper, we encourage clinicians to participate in the development of marriage education programs, such as that described by our Supporting Healthy Marriage program, which serves a large number of low-income couples, and propose a psychological conceptual framework for delivering marriage education services. It makes sense for clinicians to consider using this novel approach given the opportunity to impact such a large segment of society that might not receive psychological services.

Reasonable efforts? Implementation of the reunification bypass provision of ASFA.

The Adoption and Safe Families Act (ASFA) of 1997 includes provisions to deny reunification services under specified conditions and gives states latitude to develop any number of additional "aggravated circumstances" in which parents need not be offered services. California legislators have developed a relatively large number of conditions enabling agencies to bypass reunification services. Based upon a case record review involving 1,055 parents, this study attempts to identify the proportion of parents eligible for a reunification bypass, the proportion recommended to the courts, and the proportion of parents who were denied reunification services, and examines the characteristics of parents associated with reunification bypass recommendations. Based upon focus groups and interviews with child welfare and judicial personnel in six counties, the study also examines the implementation of reunification bypass provisions. Implications for public policy and practice are provided.

Larval feeding duration affects ecdysteroid levels and nutritional reserves regulating pupal commitment in the yellow fever mosquito Aedes aegypti (Diptera: Culicidae).

What little is known about the endocrine regulation of mosquito development suggests that models based on Lepidoptera and Drosophila may not apply. We report on basic parameters of larval development and the commitment to metamorphosis in the yellow fever mosquito Aedes aegypti that are affected by varying the length of feeding time for last instar larvae. A critical mass for pupal commitment was achieved after 24 h of feeding by last instars, also the age at which tissue production and hemolymph titers of ecdysteroids are increasing. A greater proportion of last instars successfully pupated and eclosed as adults as the length of their feeding time increased. Less than 24 h of feeding time resulted in last instars that were developmentally arrested; these larvae tolerated starvation conditions for up to 2 weeks and retained the capacity to pupate if re-fed. Starvation tolerance may be a common trait among container-inhabiting species, and this period is an important factor to be considered for vectorial capacity and control measures. To distinguish cues for metamorphosis related to a larva's nutritional status versus its age, newly molted last instars were fed for different periods of time but sampled at the same age; ecdysteroid levels, body mass and nutrient reserves were then measured for each group. Our data suggest that metamorphic capacity is dependent on a larva's nutritional condition and not just the age at which ecdysteroid titers increase. Last instars that have fed for a particular length of time may initiate their metamorphic molt when both threshold levels of nutrient reserves and ecdysteroid titer have been met. Future studies will lead to a conceptual model specific for the nutritional and hormonal regulation of mosquito post-embryonic development. This model should facilitate the exploitation of current and novel insect growth regulators that are among favored strategies for vector population suppression.