A bispecific anti-PD-1 and PD-L1 antibody induces PD-1 cleavage and provides enhanced anti-tumor activity.

Combinatorial strategies, such as targeting different immune checkpoint receptors, hold promise to increase the breadth and duration of the response to cancer therapy. Here we describe the preclinical evaluation of CTX-8371, a protein construct which combines PD-1 and PD-L1 targeting in one bispecific, tetravalent antibody. CTX-8371 matched or surpassed the activity of anti-PD-1 and PD-L1 benchmark antibodies in several in vitro T cell activation assays and outperformed clinically approved benchmarks in the subcutaneous MC38 colon and the B16F10 lung metastasis mouse tumor models. Investigation into the mechanism of action revealed that CTX-8371 co-engagement of PD-1 and PD-L1 induced the proteolytic cleavage and loss of cell surface PD-1, which is a novel and non-redundant mechanism that adds to the PD-1/PD-L1 signaling axis blockade. The combination of CTX-8371 and an agonistic anti-CD137 antibody further increased the anti-tumor efficacy with long-lasting curative therapeutic effect. In summary, CTX-8371 is a novel checkpoint inhibitor that might provide greater clinical benefit compared to current anti-PD-1 and PD-L1 antibodies, especially when combined with agents with orthogonal mechanisms of action, such as agonistic anti-CD137 antibodies.

Optimizing Type 2 Diabetes Management in a Medically Complex Patient: A Case Report of a Patient with Type 2 Diabetes and HIV.

Background: The prevalence of diabetes is rapidly escalating, with projections indicating that 783 million individuals aged 20-79 years worldwide will be affected by diabetes. This rise is concurrent with a persistent prevalence of HIV in developing nations, while conventional risk factors such as sedentary lifestyle and unhealthy diet may account for this trend, HIV and its treatment have emerged as potential contributing factors. Achieving optimal diabetes control in patients with HIV necessitates a profound understanding of the intricate interplay between the two diseases and their respective treatments. Case Report: We present a case involving a patient with long standing type 2 diabetes, coexisting HIV infection and hypertension. Despite receiving high doses of insulin, as advised by most diabetes guidelines, the patient's diabetes remained poorly controlled. In lieu of strictly adhering to guidelines, our primary focus was to conduct a comprehensive reevaluation of the patient's medications, prioritizing the development of streamlined and safe treatment regimens for all three of her medical conditions. Employing this strategy, we observed swift improvement in blood glucose levels, leading to successful diabetes control within one year. Conclusion: This case underscores the importance of individualizing diabetes management in patients with multiple comorbidities. It highlights the significance of reassessing treatment approaches beyond standard guidelines, with a focus on tailoring therapy to suit the unique needs and complexities of each patient's medical profile. Such personalized interventions hold promise for achieving optimal diabetes control in individuals with diverse comorbidities.

From clinic to community: A randomized controlled trial of a peer support model for adults with type 2 diabetes from specialty care settings in British Columbia.

AIMS: To examine the impact of a 12-month peer-led diabetes self-management support intervention delivered via telephone amongst adults with type 2 diabetes (T2D) from specialty care settings in British Columbia (BC). METHODS: One-hundred ninety-six adults with T2D were randomly assigned to either a 12-month Peer-Led, Empowerment-based, Approach, to Self-management Efforts in Diabetes (PLEASED) intervention or a usual care condition. PLEASED involved weekly telephone contacts from a peer leader (PL) in the first 3 months followed by bi-weekly telephone contacts in the last 9 months. Assessments were conducted at baseline, 3 and 12 months. The primary outcome was HbA1c ; secondary outcomes included diabetes distress (DD), ApoB, systolic and diastolic blood pressure (BP), body mass index, waist circumference and depressive symptoms. RESULTS: No within or between group changes were observed for HbA1c at 3 or 12 months. However, amongst participants with HbA1c  ≥ 69 mmol/mol (8.5%), the PLEASED group significantly lowered their HbA1c at 12 months [-11.7 mmol/mol (-1.07%); 95% CI: -20.7, -2.5 (-1.89, -0.23); p = 0.016] compared to usual care. Amongst secondary outcomes, within-group improvements in overall DD were found at 3 months (-0.21; 95% CI: -0.35, -0.08; p = 0.002) for the PLEASED group and at 12 months for both groups (PLEASED: -0.35; 95% CI: -0.49, -0.21; p < 0.001 and control: -0.33; 95% CI: -0.47, -0.19; p < 0.001), however, no between-group differences were observed. The PLEASED group improved systolic BP at 12 months (-5.4 mm Hg; 95% CI: -10.0, -0.8; p = 0.023) compared to usual care. CONCLUSIONS: Participation in a peer support intervention in diabetes delivered via telephone leads to long-term improvements in HbA1c amongst high-risk adults with T2D living in BC. TRIAL REGISTRATION: The study was registered on clinicaltrials.gov (NT02804620).

Case Series of U-500 Insulin Use in Adults With Type 2 Diabetes and Severe Insulin Resistance.

OBJECTIVES: Severe insulin resistance results in large volumes of insulin to achieve glycemic control. These large volumes can result in patient discomfort and decreased satisfaction. Using the more concentrated U-500 insulin provides a solution to this problem. This case series demonstrates real-world use of U-500 insulin in a Canadian population. METHODS: Seventeen patients were identified to have been started on U-500 insulin at an endocrinology clinic in Vancouver, British Columbia, Canada. The retrospective chart review looked at patients' characteristics before starting U-500 insulin and at their 1-year follow-up appointment. RESULTS: At follow up, patients demonstrated improved glycated hemoglobin with a mean improvement of 1.6% at 1 year (p<0.05). There was a statistically significant increase in hypoglycemia (p<0.05), and, on average, patients gained 5.6 kg over the course of the year (p<0.05). There was no statistically significant change in number of units of insulin, injections, lipids, renal function or blood pressure. CONCLUSIONS: The initiation of U-500 insulin results in improved glycemic control at the cost of increased hypoglycemia and weight gain.

Study protocol and baseline sample characteristics: From clinic to community: Using peer support as a transition model for improving long-term diabetes-related health outcomes.

BACKGROUND: The objective of this randomized controlled trial is to examine the effects of a 12-month telephone-based peer-led diabetes self-management support (DSMS) intervention on long-term diabetes-related health outcomes. METHODS: In total, 197 participants with type 2 diabetes were recruited from specialty care settings (diabetes and endocrinology clinics). They were randomly assigned to 1) a 12-month Peer-Led, Empowerment-based Approach to Self-management Efforts in Diabetes (PLEASED) program where they received 12 weekly contacts from their peer supporter (PS) in the first 3 months, followed by 18 biweekly telephone support contacts over the last 9 months, or 2) usual care. The primary clinical and psychosocial outcomes were HbA1c and diabetes distress (DD), respectively. Secondary outcomes were cardiovascular risk factors. Assessments were conducted at baseline, 3 months, and 12 months. RESULT: Of 197 recruited participants, 49.7% were female. The majority of participants were married/partnered, well-educated, employed, and Caucasian, with a mean HbA1c of 8.09 ±â€¯1.7. Forty-two percent of participants reported little or no distress. There was no significant difference between the two groups. DISCUSSION: Despite evidence showing that individuals with poor glycemic control benefit the most from peer support interventions, the majority of such interventions have been designed for and implemented in community and primary care-based settings. The present study investigates a 12-month peer support model to help patients initiate and sustain effective self-management behaviors while transitioning from specialty care to a community setting. The study was completed in November 2018. The outcome data analyses are currently underway. TRIAL REGISTRATION: The study was registered on clinicaltrials.gov (NT02804620). PROTOCOL VERSION: The protocol version is 3.5.

Impact of a Cross-Kingdom Signaling Molecule of Candida albicans on Acinetobacter baumannii Physiology.

Multidrug-resistant (MDR) Acinetobacter baumannii is an opportunistic human pathogen that has become highly problematic in the clinical environment. Novel therapies are desperately required. To assist in identifying new therapeutic targets, the antagonistic interactions between A. baumannii and the most common human fungal pathogen, Candida albicans, were studied. We have observed that the C. albicans quorum-sensing molecule, farnesol, has cross-kingdom interactions, affecting the viability of A. baumannii. To gain an understanding of its mechanism, the transcriptional profile of A. baumannii exposed to farnesol was examined. Farnesol caused dysregulation of a large number of genes involved in cell membrane biogenesis, multidrug efflux pumps (AcrAB-like and AdeIJK-like), and A. baumannii virulence traits such as biofilm formation (csuA, csuB, and ompA) and motility (pilZ and pilH). We also observed a strong induction in genes involved in cell division (minD, minE, ftsK, ftsB, and ftsL). These transcriptional data were supported by functional assays showing that farnesol disrupts A. baumannii cell membrane integrity, alters cell morphology, and impairs virulence characteristics such as biofilm formation and twitching motility. Moreover, we showed that A. baumannii uses efflux pumps as a defense mechanism against this eukaryotic signaling molecule. Owing to its effects on membrane integrity, farnesol was tested to see if it potentiated the activity of the membrane-acting polymyxin antibiotic colistin. When coadministered, farnesol increased sensitivity to colistin for otherwise resistant strains. These data provide mechanistic understanding of the antagonistic interactions between diverse pathogens and may provide important insights into novel therapeutic strategies.

Evaluation of the International Association of the Diabetes In Pregnancy Study Group new criteria: gestational diabetes project.

OBJECTIVE: To examine the diagnostic rates of gestational diabetes (GDM) and maternal/fetal outcomes before and after replacement of the Carpenter and Coustan (C&C) criteria with the International Association Diabetes Pregnancy Study Group (IADPSG) criteria. METHODS: A retrospective analysis of all pregnancies in 2 separate 6-month cohorts in the province of British Columbia. The first C&C cohort was defined as a 6-month period prior to the introduction of the IADPSG 75 g glucose tolerance test on October 1, 2010. The IADPSG cohort was studied during a 6-month period after the change. RESULTS: There was a significant increase in rates of GDM when using the IADPSG 75 g criteria, from 7.9% (1838 of 23 211) to 9.4% (2104 of 22 397). There were no significant changes in maternal outcomes when using the IADPSG criteria (caesarean section, induction of labour, perineal laceration, pregnancy-induced hypertension, antepartum hemorrhage >20 weeks or postpartum hospital length of stay). The caesarean section rate was not increased according to multivariate analysis (30.9% vs. 29.7%; p=0.073). There were no significant changes in most fetal outcomes when using the IADPSG criteria (mean gestational age at birth, premature birth, meconium, birth trauma, mean birth weight, large for gestational age, small for gestational age, intrauterine growth restriction), but neonatal hypoglycemia was significantly higher (1.6 % vs. 1.3 %; p=0.007). CONCLUSIONS: The rates of GDM were higher when using the new IADPSG criteria. Overall, all of the maternal and most of the fetal outcomes were similar.

Insulin pump use in pregnancy is associated with lower HbA1c without increasing the rate of severe hypoglycaemia or diabetic ketoacidosis in women with type 1 diabetes.

AIMS/HYPOTHESIS: The aim of this study was to compare glycaemic control and maternal-fetal outcomes in women with type 1 diabetes managed on insulin pumps compared with multiple daily injections of insulin (MDI). METHODS: In a retrospective study, glycaemic control and outcomes of 387 consecutive pregnancies in women with type 1 diabetes who attended specialised clinics at three centres 2006-2010 were assessed. RESULTS: Women using insulin pumps (129/387) were older and had a longer duration of diabetes, more retinopathy, smoked less in pregnancy, and had more preconception care (p < 0.01 for each). Among 113 pregnancies >20 weeks' gestation in women on insulin pumps and 218 in women on MDI, there was a significant difference in HbA1c in the first trimester (mean HbA1c 6.90 ± 0.71% (52 ± 7.8 mmol/mol) vs 7.60 ± 1.38% (60 ± 15.1 mmol/mol), p < 0.001), which persisted until the third trimester (mean HbA1c 6.49 ± 0.52% (47 ± 5.7 mmol/mol) vs 6.81 ± 0.85% (51 ± 9.3 mmol/mol), p = 0.002). Rates of diabetic ketoacidosis were similar in women on insulin pumps vs MDI (1.8% vs 3.0%, p = 0.72). Despite lower HbA1c, women on insulin pumps did not have an increased incidence of severe hypoglycaemia (8.0% vs 7.6%, p = 0.90) or more weight gain (16.3 ± 8.7 vs 15.2 ± 6.2 kg, p = 0.18). More large-for-gestational-age infants in the pump group (55.0% vs 39.2%, p = 0.007) may have resulted from confounding by parity. CONCLUSIONS/INTERPRETATION: In this large multicentre study, women using insulin pumps in pregnancy had lower HbA1c without increased risk of severe hypoglycaemia or diabetic ketoacidosis but no improvement in other pregnancy outcomes. This information can help inform care providers and patients about the glycaemic effectiveness and safety of insulin pumps in pregnancy.

The prevalence of gestational diabetes mellitus and glucose abnormalities in pregnant women with hepatitis C virus infection in British Columbia.

OBJECTIVE: There is evidence to support an association between diabetes mellitus (DM) and hepatitis C virus (HCV) infection. The insulin resistant state of pregnancy suggests a predisposition to developing gestational diabetes mellitus (GDM) in women infected with HCV. The aim of this study was to compare the prevalence of GDM and impaired glucose tolerance (IGT) of pregnancy between women infected with HCV and the general population of British Columbia screened for GDM. METHODS: The HCV cohort was drawn from a population-based prospective cohort of 148 pregnant women infected with HCV in British Columbia. GDM screening tests were completed in 84 women. The prevalence of GDM and IGT of pregnancy in the general population of British Columbia was estimated by acquiring 24 321 GDM screening tests performed by the two major laboratories in the province. RESULTS: Non-compliance was the primary reason for incomplete screening. The prevalence of GDM was 9.5% in the HCV cohort and 6.8% in the screened general population (chi-square test P = 0.33). Similarly, there was no difference in IGT of pregnancy between the two cohorts (2.4% vs. 3.5%; chi-square test P = 0.57). CONCLUSION: A difference in the prevalence of either GDM or IGT of pregnancy was not detected between HCV-infected patients who were screened for GDM and those screened in the general population. Further studies are required to assess whether HCV infection is an independent risk factor for GDM.

Probing the catalytic activity and heterogeneity of Au-nanoparticles at the single-molecule level.

Nanoparticles can catalyze many important chemical transformations in organic synthesis, pollutant removal, and energy production. Characterizing their catalytic properties is essential for understanding the fundamental principles governing their activities, but is challenging in ensemble measurements due to their intrinsic heterogeneity from their structural dispersions, heterogeneous surface sites, and surface restructuring dynamics. To remove ensemble averaging, we recently developed a single-particle approach to study the redox catalysis of individual Au-nanoparticles in solution. By detecting the fluorescence of the catalytic product at the single-molecule level, we followed the catalytic turnovers of single Au-nanoparticles in real time at single-turnover resolution. Here we extend our single-nanoparticle studies to examine in detail the activity and heterogeneity of 6 nm spherical Au-nanoparticles. By analyzing the statistical properties of single-particle reaction waiting times across a range of substrate concentrations, we directly determine the distributions of kinetic parameters of individual Au-nanoparticles, including the rate constants and the equilibrium constants of substrate adsorption, and quantify their heterogeneity. Large activity heterogeneity is observed among the Au-nanoparticles in both the catalytic conversion reaction and the product dissociation reaction, which are typically hidden in ensemble-averaged measurements. Analyzing the temporal fluctuation of catalytic activity of individual Au-nanoparticles further reveals that these nanoparticles have two types of surface sites with different catalytic properties-one type-a with lower activity but higher substrate binding affinity, and the other type-b with higher activity but lower substrate binding affinity. Each Au-nanoparticle exhibits type-a behavior at low substrate concentrations and switches to type-b behavior at a higher substrate concentration, and the switching concentration varies largely from one nanoparticle to another. The heterogeneous and dynamic behavior of Au-nanoparticle catalysts highlight the intricate interplay between catalysis, structural dispersion, variable surface sites, and surface restructuring dynamics in nanocatalysis.

Single-molecule nanocatalysis reveals heterogeneous reaction pathways and catalytic dynamics.

Nanoparticles are important catalysts for many chemical transformations. However, owing to their structural dispersions, heterogeneous distribution of surface sites and surface restructuring dynamics, nanoparticles are intrinsically heterogeneous and challenging to characterize in ensemble measurements. Using a single-nanoparticle single-turnover approach, we study the redox catalysis of individual colloidal Au nanoparticles in solution, using single-molecule detection of fluorogenic reactions. We find that for product generation, all Au nanoparticles follow a Langmuir-Hinshelwood mechanism but with heterogeneous reactivity; and for product dissociation, three nanoparticle subpopulations are present that show heterogeneous reactivity between multiple dissociation pathways with distinct kinetics. Correlation analyses of single-turnover waiting times further reveal activity fluctuations of individual Au nanoparticles, attributable to both catalysis-induced and spontaneous dynamic surface restructuring that occurs at different timescales at the surface catalytic and product docking sites. The results exemplify the power of the single-molecule approach in revealing the interplay of catalysis, heterogeneous reactivity and surface structural dynamics in nanocatalysis.

Engineered holliday junctions as single-molecule reporters for protein-DNA interactions with application to a MerR-family regulator.

Protein-DNA interactions are essential for gene maintenance, replication, and expression. Characterizing how proteins interact with and change the structure of DNA is crucial in elucidating the mechanisms of protein function. Here, we present a novel and generalizable method of using engineered DNA Holliday junctions (HJs) that contain specific protein-recognition sequences to report protein-DNA interactions in single-molecule FRET measurements, utilizing the intrinsic structural dynamics of HJs. Because the effects of protein binding are converted to the changes in the structure and dynamics of HJs, protein-DNA interactions that involve small structural changes of DNA can be studied. We apply this method to investigate how the MerR-family regulator PbrR691 interacts with DNA for transcriptional regulation. Both apo- and holo-PbrR691 bind the stacked conformers of the engineered HJ, change their structures, constrain their conformational distributions, alter the kinetics, and shift the equilibrium of their structural dynamics. The information obtained maps the potential energy surfaces of HJ before and after PbrR691 binding and reveals the protein actions that force DNA structural changes for transcriptional regulation. The ability of PbrR691 to bind both HJ conformers and still allow HJ structural dynamics also informs about its conformational flexibility that may have significance for its regulatory function. This method of using engineered HJs offers quantification of the changes both in structure and in dynamics of DNA upon protein binding and thus provides a new tool to elucidate the correlation of structure, dynamics, and function of DNA-binding proteins.

Normal perinatal mortality in type 1 diabetes mellitus in a series of 300 consecutive pregnancy outcomes.

Reports of outcomes of pregnancy in women with type 1 diabetes have consistently found increased perinatal mortality and morbidity. The primary objective of our study was to compare the perinatal mortality rate in type 1 diabetic pregnancies with that of the general population. The secondary objective was to compare the morbidities in these groups. A series of 247 women with type 1 diabetes had 300 consecutive pregnancy outcomes analyzed over a 10-year period. They were compared with the control population from the same hospital. Perinatal mortality was 6.6/1000 (95% CI, 0-17), which was significantly lower than the control population rate of 31/1000. There was an increased incidence of morbidity including maternal hypertension, cesarean section, preterm delivery, birth injury, large for gestational age infants, admissions to neonatal intensive care, neonatal hypoglycemia, and phototherapy. Pregnancies in type 1 diabetes can be associated with a normal perinatal mortality rate although morbidity remains elevated compared with controls.