Understanding the delivery of substance use treatment services to transgender and gender-diverse people: Findings from a mixed-methods study of healthcare professionals.

BACKGROUND: Transgender and gender diverse (TGD) people who use drugs report barriers to accessing substance use treatment, including provider mistreatment. Little research has explored the multilevel factors that shape the capacity of substance use treatment professionals to provide gender-affirmative care (i.e., care that respects and affirms one's gender) to TGD people. METHODS: From October 2021 to March 2022, substance use treatment and harm reduction professionals in Rhode Island were surveyed (N = 101) and qualitatively interviewed (N = 19) about the provision of substance use treatment-related services to TGD people. Quantitative data were analyzed descriptively; differences were examined using Fisher exact tests (p < 0.05). Qualitative interviews were coded and analyzed using thematic analysis. RESULTS: Participants reported limited exposure to TGD people and lacked training on TGD health, which resulted in limited cultural and clinical competency and low self-efficacy in their ability to care for TGD people. Participants also highlighted structural factors (e.g., non-inclusive intake forms, limited availability of gender-inclusive ancillary community services) that restricted their ability to provide effective and affirming care to TGD people. Some participants also reported a "gender blind" ethos at their institutions- described by some as ignoring the potential impact of TGD peoples' unique experiences on their substance use and ability to benefit from treatment. While some perceived gender blindness as problematic, others believed this approach enabled substance use treatment professionals to consider all the identities and needs that patients/clients may have. Despite differences in treatment approaches, most participants agreed that their workplaces could benefit from efforts to create a safe and affirming space for people who use drugs, particularly TGD patients/clients. CONCLUSION: Results underscore how structural, interpersonal, and individual factors contributed to barriers in the provision of gender-affirmative substance use-related care for TGD people. Findings can inform efforts to increase the capacity of providers to deliver gender-affirmative substance use-related services, which is essential to supporting the recovery goals of TGD people.

Alpaca Field Behaviour When Cohabitating with Lambing Ewes.

A common strategy to reduce predator attack on livestock is the deployment of guardian alpacas. However, little research has been conducted on the behaviour of this species while housed with other livestock. This study monitored two male alpacas cohabitating with 180 lambing ewes in order to quantify field behaviour in two phases. Phase one assessed diurnal patterns of alpacas and lambing ewes using Global Navigation Satellite System (GNSS) collars recording data over 41 days, in combination with observational recordings. Phase two developed an alpaca behavioural ethogram through continuous observations from 05:30 to 19:30 h over a 3-day period. The two alpacas shared similar behaviours with commonality of distance travelled, and both species exhibited an increase in activity level based on speed between the times of 05:00 and 17:00 h. The GNSS data indicated that the alpacas flocked with the ewes at night sharing the same resting location, however, would spend time during the day on the outskirts of the paddock. Alpacas were observed to spend the majority of the observation period in two behavioural states: grazing (57%) and resting (27%). As a result of this study we were able to catalogue a range and frequency of field behaviours which alpacas exhibit while cohabitating with lambing ewes. However, further research is needed to determine in more detail how these behaviours correspond with the effectiveness of this species as a livestock guardian.

Engineering of Bacillus subtilis strains to allow rapid characterization of heterologous diguanylate cyclases and phosphodiesterases.

Microbial processes, including biofilm formation, motility, and virulence, are often regulated by changes in the available concentration of cyclic dimeric guanosine monophosphate (c-di-GMP). Generally, high c-di-GMP concentrations are correlated with decreased motility and increased biofilm formation and low c-di-GMP concentrations are correlated with an increase in motility and activation of virulence pathways. The study of c-di-GMP is complicated, however, by the fact that organisms often encode dozens of redundant enzymes that synthesize and hydrolyze c-di-GMP, diguanylate cyclases (DGCs), and c-di-GMP phosphodiesterases (PDEs); thus, determining the contribution of any one particular enzyme is challenging. In an effort to develop a facile system to study c-di-GMP metabolic enzymes, we have engineered a suite of Bacillus subtilis strains to assess the effect of individual heterologously expressed proteins on c-di-GMP levels. As a proof of principle, we characterized all 37 known genes encoding predicted DGCs and PDEs in Clostridium difficile using parallel readouts of swarming motility and fluorescence from green fluorescent protein (GFP) expressed under the control of a c-di-GMP-controlled riboswitch. We found that 27 of the 37 putative C. difficile 630 c-di-GMP metabolic enzymes had either active cyclase or phosphodiesterase activity, with agreement between our motility phenotypes and fluorescence-based c-di-GMP reporter. Finally, we show that there appears to be a threshold level of c-di-GMP needed to inhibit motility in Bacillus subtilis.

Functional characterization of core components of the Bacillus subtilis cyclic-di-GMP signaling pathway.

Bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) is an intracellular second messenger that regulates adaptation processes, including biofilm formation, motility, and virulence in Gram-negative bacteria. In this study, we have characterized the core components of a c-di-GMP signaling pathway in the model Gram-positive bacterium Bacillus subtilis. Specifically, we have directly identified and characterized three active diguanylate cyclases, DgcP, DgcK, and DgcW (formerly YtrP, YhcK, and YkoW, respectively), one active c-di-GMP phosphodiesterase, PdeH (formerly YuxH), and a cyclic-diguanylate (c-di-GMP) receptor, DgrA (formerly YpfA). Furthermore, elevation of c-di-GMP levels in B. subtilis led to inhibition of swarming motility, whereas biofilm formation was unaffected. Our work establishes paradigms for Gram-positive c-di-GMP signaling, and we have shown that the concise signaling system identified in B. subtilis serves as a powerful heterologous host for the study of c-di-GMP enzymes from bacteria predicted to possess larger, more-complex signaling systems.