Search

The recombinant, live, attenuated, tetravalent dengue vaccine CYD-TDV has shown efficacy against all four dengue serotypes. In this exploratory study (CYD59, NCT02827162), we evaluated potential associations of host human leukocyte antigen (HLA) alleles with dengue antibody responses, CYD-TDV vaccine efficacy, and virologically-confirmed dengue (VCD) cases. Children 4-11 years old, who previously completed a phase 2b efficacy study of CYD-TDV in a single center in Thailand, were included in the study. Genotyping of HLA class I and II loci was performed by next-generation sequencing from DNA obtained from 335 saliva samples. Dengue neutralizing antibody titers (NAb) were assessed as a correlate of risk and protection. Regression analyses were used to assess associations between HLA alleles and NAb responses, vaccine efficacy, and dengue outcomes. Month 13 NAb log geometric mean titers (GMTs) were associated with decreased risk of VCD. In the vaccine group, HLA-DRB1*11 was significantly associated with higher NAb log GMT levels (beta: 0.76; p = 0.002, q = 0.13). Additionally, in the absence of vaccination, HLA associations were observed between the presence of DPB1*03:01 and increased NAb log GMT levels (beta: 1.24; p = 0.005, q = 0.17), and between DPB1*05:01 and reduced NAb log GMT levels (beta: -1.1; p = 0.001, q = 0.07). This study suggests associations of HLA alleles with NAb titers in the context of dengue outcomes. This study was registered with clinicaltrials.gov: NCT02827162.

Objectives: Patients with ovarian cancer have been shown to have high rates of venous thromboembolism (VTE), with previous studies reporting an up to 16% overall incidence of symptomatic VTE, and a 7.5% cumulative incidence of VTE within the first 30 days after surgery. The goal of this study is to determine the updated incidence and risk factors for VTE for patients with advanced epithelial ovarian cancer undergoing first-line therapy, including cytoreductive surgery, on an Enhanced Recovery After Surgery (ERAS) protocol. Methods: Medical records were reviewed for all patients with FIGO stage IIIA-IVB epithelial ovarian, fallopian tube, or primary peritoneal cancer undergoing primary or interval cytoreductive surgery from March 2017 through September 2019 at a single institution. All patients were enrolled on an ERAS protocol, including 28-day postoperative VTE prophylaxis. Demographic information, past medical history, perioperative characteristics, and ERAS compliance were collected and evaluated using univariate and multivariate Cox proportional hazards models. Download : Download high-res image (86KB) Download : Download full-size image Results: Of 230 patients undergoing cytoreductive surgery via laparotomy, 155 received neoadjuvant chemotherapy and 75 received primary cytoreduction. 38 patients had a VTE during the study period. 13 events (5.7%) were identified at time of diagnosis, 6 (3.9%) during neoadjuvant chemotherapy, 5 (2.2%) within 30 days after surgery, 5 (2.2%) between 30 days and 6 months after surgery, and 9 (3.9%) after the 6-month window. The cumulative incidence of VTE was 6.1% (95% CI, 4.3-8.8%) within 6 months after diagnosis and 8.5% (6.2-11.4%) within 1 year after diagnosis. Estimated blood loss (adjusted HR 1.22 [95% CI, 1.09-1.36], p=0.001) and history of VTE (7.06 [2.34-21.29], p=0.001) were independently associated with VTE. Conclusions: With implementation of an ERAS protocol and 28-day postoperative VTE prophylaxis, only 1 in 46 patients experienced a VTE within 30 days after surgery, a rate markedly lower than historically reported for cytoreductive procedures. However, overall VTE occurred in 1 in 16 patients during first-line therapy. Half of the venous thromboembolic events occurred prior to surgery and 36.8% occurred more than 30 days after surgery. Strategies to further reduce VTE risk, especially during neoadjuvant chemotherapy and surveillance, should be investigated.

A long-acting injectable formulation of rilpivirine (RPV), a non-nucleoside reverse transcriptase inhibitor, is currently under investigation for use in human immunodeficiency virus (HIV) maintenance therapy. We previously characterized RPV metabolism after oral dosing and identified seven metabolites: four metabolites resulting from mono- or dioxygenation of the 2,6-dimethylphenyl ring itself or either of the two methyl groups located on that ring, one N-linked RPV glucuronide conjugate, and two O-linked RPV glucuronides produced via glucuronidation of mono- and dihydroxymethyl metabolites. However, as is true for most drugs, the metabolism of RPV after injection has yet to be reported. The phase II clinical trial HPTN 076 enrolled 136 HIV-uninfected women and investigated the safety and acceptability of long-acting injectable RPV for use in HIV pre-exposure prophylaxis. Through the analysis of plasma samples from 80 of these participants in the active product arm of the study, we were able to detect 2 metabolites after intramuscular injection of long-acting RPV, 2-hydroxymethyl-RPV, and RPV N-glucuronide. Of the total of 80 individuals, 72 participants exhibited detectable levels of 2-hydroxymethyl-RPV in plasma samples whereas RPV N-glucuronide was detectable in plasma samples of 78 participants. In addition, RPV N-glucuronide was detectable in rectal fluid, cervicovaginal fluid, and vaginal tissue. To investigate potential genetic variation in genes encoding enzymes relevant to RPV metabolism, we isolated genomic DNA and performed next-generation sequencing of CYP3A4, CYP3A5, UGT1A1 and UGT1A4. From these analyses, four missense variants were detected for CYP3A4 whereas one missense variant and one frameshift variant were detected for CYP3A5. A total of eight missense variants of UGT1A4 were detected, whereas two variants were detected for UGT1A1; however, these variants did not appear to account for the observed interindividual variability in metabolite levels. These findings provide insight into the metabolism of long-acting RPV and contribute to an overall understanding of metabolism after oral dosing versus injection. ClinicalTrials.gov Identifier: NCT02165202

Cabotegravir (CAB) is an integrase strand-transfer inhibitor of HIV that has proven effective for HIV treatment and prevention in a long-acting injectable formulation, typically preceded by an oral formulation lead-in phase. Previous in vitro studies have demonstrated that CAB is primarily metabolized via glucuronidation by uridine diphosphate glucuronosyltransferase (UGT) 1A1 and 1A9. In this study, we performed next-generation sequencing of genomic DNA isolated from the HPTN 077 participants to explore the variants within UGT1A1 and UGT1A9. Additionally, to enable correlation of UGT1A1 and UGT1A9 genotypes with plasma CAB-glucuronide levels, we quantified glucuronidated CAB following both oral administration of CAB and intramuscular injection of long-acting CAB. From these studies, 48 previously unreported variants of UGT1A1 and UGT1A9 were detected. Notably, 5/68 individuals carried a UGT1A1 454C>A variant that resulted in amino acid substitution P152T, and the use of in silico tools predicted a deleterious effect of the P152T substitution. Thus, the impact of this mutant on a range of UGT1A1 substrates was tested using a COS-7 cell-based assay. The glucuronide conjugates of CAB, dolutegravir, and raltegravir, were not formed in the COS-7 cells expressing the UGT1A1 P152T mutant. Further, formation of glucuronides of raloxifene and 7-ethyl-10-hydroxycamptothecin were reduced in the cells expressing the UGT1A1 P152T mutant. Using the same approach, we tested the activities of two UGT1A9 mutants, UGT1A9 H217Y and UGT1A9 R464G, and found that these mutations were tolerated and decreased function, respectively. These data provide insight into previously unreported genetic variants of UGT1A1 and UGT1A9.

Summary Background Long-acting injectable cabotegravir is a novel integrase inhibitor currently in advanced clinical development for HIV prevention and treatment. We aimed to assess the terminal phase pharmacokinetics and safety of long-acting injectable cabotegravir in participants included in the HPTN 077 trial. Methods HPTN 077 was a multicentre, double-blind, randomised, placebo-controlled phase 2a trial done at eight sites in Brazil, Malawi, South Africa, and the USA. Participants (aged 18–65 years), who were HIV-uninfected and at low-risk for HIV, were randomly assigned (3:1) to long-acting injectable cabotegravir (800 mg given three times at 12 week intervals or 600 mg given five times, administered at one 4 week interval, and every 8 weeks thereafter) or placebo. Participants were followed up to 76 weeks after final injection. In a prespecified analysis of secondary and exploratory outcomes, we assessed the safety, measured by the proportion of participants with grade 2 or worse adverse events, and pharmacokinetics, measured by apparent terminal phase half-life (t1/2app) and estimated time to lower limit of quantification (LLOQ) of long-acting injectable cabotegravir during the injection phase (defined as the time between first injection and 12 weeks or 8 weeks after the last injection in cohort 1 or cohort 2 respectively) and tail phase (defined as the time between final injection and 52–76 weeks post-final injection). Safety was analysed in all participants who received at least one injection. Pharmacokinetic analyses included all participants who had received at least one injection and had at least three cabotegravir measurements higher than the LLOQ after the final injection. Pharmacokinetic outcomes were estimated using non-compartmental methods. The trial is completed, and was registered with ClinicalTrials.gov , NCT02178800 . Findings Between Feb 9, 2015, and May 27, 2016, 177 participants (134 participants in the cabotegravir group [74 participants in cohort 1; 60 participants in cohort 2] and 43 participants in the placebo group [25 participants in cohort 1; 18 participants in cohort 2) were enrolled and received at least one injection and thus were included in the safety analysis. The incidence of grade 2 or worse adverse events was significantly lower during the tail phase than the injection phase (p Interpretation The clinical significance of the long pharmacokinetic tail of cabotegravir observed in female participants compared with male participants, and those with higher BMI compared with a lower BMI, need to be addressed in future trials. Funding National Institute of Allergy and Infectious Diseases.

ObjectivesOvarian cancer patients with miliary disease have the lowest rates of complete surgical resection and poorest survival. Adjunct surgical techniques may potentially increase rates of complete surgical resection. No studies have evaluated the use of these techniques in primary debulking surgery for ovarian cancer patients with miliary disease. The aim of this study was to examine the use of adjunct surgical techniques during primary debulking surgery for patients with advanced epithelial ovarian, fallopian tube, and primary peritoneal cancer with miliary disease.MethodsMedical records of patients with International Federation of Gynecology and Obstetrics (FIGO) stages IIIC–IVB epithelial ovarian, fallopian tube, or primary peritoneal cancer with miliary disease undergoing primary debulking surgery from January 2010 to December 2014 were reviewed. Adjunct surgical techniques were defined as ultrasonic surgical aspiration, argon enhanced electrocautery, thermal plasma energy, and traditional electrocautery ablation. Patients undergoing surgery with and without these devices were compared with respect to demographics, operative characteristics, postoperative complications, residual disease, progression free survival and overall survival.ResultsA total of 135 patients with miliary disease underwent primary debulking surgery, of which 30 (22.2%) patients used adjunct surgical techniques. The most common devices were ultrasonic surgical aspiration (40%) and argon enhanced electrocautery (36.7%). The most common sites of use were diaphragm (63.3%), pelvic peritoneum (30%), bowel mesentery (20%), and large bowel serosa (20%). There were no differences in age, stage, primary site, histology, operative time, surgical complexity, or postoperative complications for patients operated on with or without these devices. Volume of residual disease was similar (0.1–1 cm: 60% with adjunct techniques versus 68.6% without; complete surgical resection: 16.7% with adjunct techniques versus 13.3% without; p=0.67). For patients with ≤1 cm residual disease, median progression free survival (15 versus 15 months, p=0.65) and median overall survival (40 versus 55 months, p=0.38) were also similar.ConclusionAdjunct surgical techniques may be incorporated during primary debulking surgery for patients with advanced epithelial ovarian, fallopian tube, or primary peritoneal cancer with miliary disease; however, these do not improve the rate of optimal cytoreduction.

OBJECTIVE: To compare gynecologic oncology surgical treatment modifications and delays during the first wave of the COVID-19 pandemic between a publicly funded Canadian versus a privately funded American cancer center. METHODS: This is a retrospective cohort study of all planned gynecologic oncology surgeries at University Health Network (UHN) in Toronto, Canada and Brigham and Women's Hospital (BWH) in Boston, USA, between March 22,020 and July 302,020. Surgical treatment delays and modifications at both centers were compared to standard recommendations. Multivariable logistic regression was performed to adjust for confounders. RESULTS: A total of 450 surgical gynecologic oncology patients were included; 215 at UHN and 235 at BWH. There was a significant difference in median time from decision-to-treat to treatment (23 vs 15 days, p < 0.01) between UHN and BWH and a significant difference in treatment delays (32.56% vs 18.29%; p < 0.01) and modifications (8.37% vs 0.85%; p < 0.01), respectively. On multivariable analysis adjusting for age, race, treatment site and surgical priority status, treatment at UHN was an independent predictor of treatment modification (OR = 9.43,95% CI 1.81-49.05, p < 0.01). Treatment delays were higher at UHN (OR = 1.96,95% CI 1.14-3.36 p = 0.03) and for uterine disease (OR = 2.43, 95% CI 1.11-5.33, p = 0.03). CONCLUSION: During the first wave of COVID-19 pandemic, gynecologic oncology patients treated at a publicly funded Canadian center were 9.43 times more likely to have a surgical treatment modification and 1.96 times more likely to have a surgical delay compared to an equal volume privately funded center in the United States.

Oncogene-induced senescence (OIS) is a well-described autonomous tumour suppressor mechanism which removes cells harbouring oncogenic mutations from the proliferation pool. However, senescent cells remain metabolically active and express factors of the senescence-associated secretory phenotype (SASP), which can conversely have a pro-tumorigenic impact on the cellular microenvironment. Yet, aberrant oncogene activation alone is insufficient for triggering OIS, as studies both in vitro and in vivo have demonstrated that low-dose oncogene cells continue proliferating, in some cases proceeding to full malignancy. The molecular mechanisms governing this dose-dependent switch remain unclear. To address this question, we developed a cell system where a constitutively active fluorescence marker (mVenus) stoichiometrically reflects the level of inducible oncogenic HRASG12V. This allows pre-induction sorting into subpopulations expressing differing levels of mVenus, which is reflected in differences in HRAS dose following induction. Using this system, we first validated that subpopulations expressing different levels of RAS are distinct in their proliferative / senescence phenotypes, implying a theoretical, cell-autonomous ‘oncogenic tipping point’ above which cells enter into OIS. We then demonstrate the potential utility of our proposed ‘predictive reporter’ system in addressing gene dose-related biological questions. RNA sequencing of sorted subpopulations demonstrates the effect of RAS dose on gene expression, where between-subpopulation differences suggests that this sorting strategy may serve as a viable intermediate between bulk and single-cell sequencing technologies. These global dose-dependent differences were validated using single-cell RNA sequencing, which also demonstrated transcriptomic heterogeneity that exists independently of RAS dose. Finally, we demonstrate that the reconstruction of subpopulations into a mixed population with cells of varying RAS doses can have implications on phenotype, and that the current system is a flexible tool that facilitates these sorts of ‘cellular ecosystem’ type experiments in vitro.

The effect of non-injection substance use on HIV viral load (VL) is understudied in international settings. Data are from HPTN063, a longitudinal observational study of HIV-infected individuals in Brazil, Thailand, and Zambia, with focus on men with VL data (Brazil = 146; Thailand = 159). Generalized linear mixed models (GLMM) assessed whether non-injection substance use (stimulants, cannabis, alcohol, polysubstance) was associated with VL undetectability. ART adherence and depressive symptoms were examined as mediators of the association. In Thailand, substance use was not significantly associated with VL undetectability or ART adherence, but alcohol misuse among MSM was associated with increased odds of depression (AOR = 2.75; 95% CI 1.20, 6.32, p = 0.02). In Brazil, alcohol misuse by MSM was associated with decreased odds of undetectable VL (AOR = 0.34; 95% CI 0.13, 0.92, p = 0.03). Polysubstance use by heterosexual men in Brazil was associated with decreased odds of ART adherence (AOR = 0.25; 95% CI 0.08, 0.78, p = 0.02). VL suppression appears attainable among non-injection substance users. Substance use interventions among HIV-positive men should address depression, adherence, and VL undetectability.

Author(s): Blair, Cherie S; Li, Sue; Chau, Gordon; Cottle, Leslie; Richardson, Paul; Marzinke, Mark A; Eshleman, Susan H; Adeyeye, Adeola; Rinehart, Alex R; Margolis, David; McCauley, Marybeth; Hendrix, Craig W; Landovitz, Raphael J; HPTN 077 Study Team | Abstract: ObjectivesTo evaluate whether hormonal contraceptive use among cisgender women is associated with differences in pharmacokinetic (PK) parameters of a long-acting injectable formulation of the integrase strand transfer inhibitor, cabotegravir (CAB-LA).SettingThis is a secondary analysis of 85 cisgender women enrolled in HPTN 077, a phase 2a multicenter study that enrolled HIV-uninfected, low-risk individuals in Malawi, Brazil, South Africa, and the United States.MethodsParticipants received 4-week daily oral cabotegravir lead-in, followed by CAB-LA 800 mg injection every 12 weeks (cohort 1) or 600 mg every 8 weeks (after 4-week initial interval between injections, cohort 2), over 41 weeks. Participants were followed 52-76 weeks subsequent to final injection. Generalized estimating equations and linear regression were used to evaluate differences in CAB-LA PK parameters (peak concentration, trough concentration, area under the curve, apparent terminal half-life, and time to lower limit of quantification) and self-reported hormonal contraceptive stratified by type (oral, injectable, implants, and other), controlling for body mass index and cohort.ResultsCompared to women reporting no hormonal contraception (n = 6), oral contraceptive use (n = 18) was associated with lower CAB-LA peak concentration but was not associated with differences in other PK parameters. No other hormonal contraceptive type (injectable, implants, and other) was associated with significant differences in CAB-LA PK parameters.ConclusionAlthough oral contraceptive use was associated with differences in CAB-LA peak concentration, no differences were observed in other PK parameters, suggesting that this association is not likely to be clinically significant. However, these data highlight the need for further research exploring potential drug-drug interactions between CAB-LA and hormonal contraceptives.

This population epidemiology study characterizes trends in polymerase chain reaction (PCR) test positivity for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in Washington State and the Seattle area between March 1 and April 16, 2020, before and after statewide physical distancing guidelines and stay-at-home orders.

Background: Daily oral TDF/FTC is protective against HIV infection when used for pre-exposure prophylaxis (PrEP). However, daily adherence to oral PrEP is difficult for many; therefore, finding alternative PrEP strategies remains a priority. HPTN 076 evaluated the long-acting injectable form of rilpivirine (RPV), known as RPV LA for safety, pharmacokinetics and acceptability. Methods: HPTN 076 (NTC 02165202) was a phase 2, double-blind, 2:1 randomized trial comparing the safety of 1200mg RPV LA (LA) to placebo (P). The study included a 28-day oral run-in phase of daily, self- administered oral RPV (25 mg), with directly observed oral dosing about six times. Of 136 enrolled sexually active, HIV-uninfected, low HIV-risk African (100) and US (36) adult women, injectable product was administered in two gluteal, intramuscular (IM) injections once every eight weeks to 122 participants following the oral run-in phase. A maximum of six injection time points occurred over a 48-week period. Acceptability, safety, tolerability and pharmacokinetic (PK) data were collected throughout the study. This paper includes primary endpoint data collected up to the week 52 post enrollment. Findings: The median age of the enrolled population was 31 years (IQR: 25,38), median weight 75 kg (IQR: 64, 89), median body mass index (BMI) 30 (IQR: 27, 35), 46% married, 94% Black and 60% unemployed. A total of 122 (80 LA, 42 P) women received at least one injection and 98 (64 LA, 34 P) received all six injections. During the injection phase, three women withdrew from the study (2 LA, 1 P) and 16 women discontinued study product (10 LA, 6 P). Fourteen women (11 LA and 3 P) discontinued oral study product and did not enter the injection phase. Study product discontinuations were not significantly different between the two arms throughout. Of the product discontinuations in the injection phase, 8% in LA and 5% in P arm were due to adverse events (AEs), including one randomized to the P arm with prolonged QTc interval on EKG. The proportion of women who experienced Grade 2 or higher AEs during the injection phase as the primary outcome was not significantly different between the two arms [73.8%, 95% CI: (63.2%, 82.1%) for LA and 73.8%, 95% CI: (58.9%, 84.7%), p>0.99]. Transient Grade ≥2 liver abnormalities occurred in 14% of women in the LA arm compared with 12% in P arm. Three LA women (4%) developed Grade 3 injection site reactions compared with none in P arm. In participants who received at least 1 injection, the geometric mean of overall RPV trough concentrations (Ctrough) was 62.2 ng/mL. In participants who received all six injections, the geometric mean of CTrough through the injection phase and after the last injection were 72.8 ng/mL and 100.9 ng/mL, respectively. At week 52 (eight weeks after last injection), the geometric mean of RPV Ctrough was 75.0 ng/mL. At the last injection visit (Week 44), 80 % of women who answered acceptability questions strongly agreed that they would think about using- and 68% that they would definitely use a PrEP injectable in the future. Interpretation: RPV LA IM injections every eight weeks in African and US women were safe and acceptable. Overall, despite more injection site reactions and pain in the participants receiving RPV LA the injections were well tolerated. Data from this study support the further development of injectable PrEP agents. Keyword: RPV LA as Pre exposure prophylaxis

To develop feasible probes for heavy metal ions has been considered as one of the most important research topics, due to the significance in monitoring and evaluating the toxicity of environmental water system, especially for the drinking water. This work provided a probe for Pb(II) ion detection in purified drinking water. Nickel hexacyanoferrate (NiHCF) films were prepared by vacuum evaporating nickel fine wire and transforming in a potassium ferricyanide solution on the front surface of platinum coated n-silicon. The transformed NiHCF film shows a reversible insertion electrochemistry and possesses electrocatalytic activity under illumination. The NiHCF-modified electrode was used as photoelectrode and dihydroxybenzene as photocurrent mediators to make up photoelectrochemical (PEC) sensor for Pb(II) ion detection in purified drinking water based on a two-electrode cell in absence of reference electrode and operated at zero working voltage. We have investigated the effects of Pb(II) ions on the photocurrent of hydroquinone or catechol in the purified water. The assay demonstrated good photocurrent responses by adding different concentrations of Pb(II) ion into purified drinking water with a linear range of 20–1560 nM and the limit of determination (based on S/N = 3) is 4.8 nM and 5.3 nM for the presence of hydroquinone and catechol, respectively.

Fast and accurate detection of aqueous mercury(II) ions is of significant importance as it raises serious risks for human health and the environment. Here, we report a photoelectrochemical determination of mercury(II) ions with nickel oxyhydroxide-functionalized n-silicon photoanodes at zero working voltage. Based on the catalysis effect of Hg(II) ions on the photocurrent of the catechol probe, a simple and sensitive photoelectrochemical sensor was constructed for Hg(II) ions detection in purified drinking waters (PW1, PW2 and PW3). The photocurrents show good response after the solution containing Hg(II) ions was added to the photoelectrochemically active catechol solution. Under the optimized conditions, the increase of photocurrent is proportional to the Hg(II) ions concentration, the linear dynamic range is observed from 10 nM to 450 nM with the detection limits of 5.4, 7.2 and 5.2 nM in PW1, PW2 and PW3, respectively. Our work suggests that the two electrodes structures are promising for low-cost, portable, real-time, heavy metal ion detectors.

Breast cancer is the most common cancer among women in the United States, with 266,120 estimated new cases in 2018 (Siegel et al., 2018). Ectopic breast tissue may be found anywhere along the milk line from the axilla to the vulva, although the exact histogenesis of vulvar mammary glands remains unclear. Cancers of ectopic mammary tissue are extremely rare and present unique challenges in diagnosis and management, especially in the context of concurrent breast cancer. We present a case of mammary-like adenocarcinoma of the vulva in a patient with concurrent breast and endometrial cancers. The diagnostic approach to concurrent breast, endometrial, and vulvar cancers is discussed.

Long-acting injectable PrEP could offer an alternative to daily oral PrEP, improve adherence and protection, if found acceptable, safe and effective. HPTN 077 evaluated injectable cabotegravir safety, tolerability and pharmacokinetics among HIV-uninfected males and females in sequentially-enrolled cohorts of two dosing strategies. We compared acceptability of product attributes, prevention preferences and future interest in injectable PrEP (FIIP) by region, sex-at-birth, arm and cohort and used multivariable analysis to identify FIIP determinants. Baseline injectable PrEP preferences were higher in non-U.S. sites and increased in both regions over time. In multivariable models, FIIP was most strongly associated with acceptability of product attributes, was higher in non-U.S. sites and more altruistic participants. Treatment arm and report of pain were not associated with FIIP. Injectable acceptability was highest in non-U.S. sites. Preferences for injectable versus other PrEP methods were higher among U.S. males than females, but higher among males and females in non-U.S. settings. Electronic supplementary material The online version of this article (10.1007/s10461-020-02808-2) contains supplementary material, which is available to authorized users.

Highly soluble and stable proteins are desirable for many different applications, from basic science to reaching a cancer patient in the form of a biological drug. For X-ray crystallography-where production of a protein crystal might take weeks and even months-a stable protein sample of high purity and concentration can greatly increase the chances of producing a well-diffracting crystal. For a patient receiving a specific protein drug, its safety, efficacy, and even cost are factors affected by its solubility and stability. Increased protein expression and protein stability can be achieved by randomly altering the coding sequence. As the number of mutants generated might be overwhelming, a powerful protein expression and stability screen is required. In this chapter, we describe a colony filtration technology, which allows us to screen random mutagenesis libraries for increased thermal stability-the Hot CoFi blot. We share how to create the random mutagenesis library, how to perform the Hot CoFi blot, and how to identify more thermally stable clones. We use the Tobacco Etch Virus protease as a target to exemplify the procedure.

Objectives: The major shift in health-care resource utilization during the COVID-19 pandemic to support patients requiring mechanical ventilation and intensive care monitoring has led to unprecedented cancellations of elective surgeries and reductions of ambulatory clinic visits worldwide. The primary objective of this study is to determine whether the response to the pandemic resulted in modifications, cancellations or delays to the standard therapeutic algorithms for patients with gynecologic malignancies at tertiary, large-volume publicly funded Canadian cancer centers as compared to a privately funded American cancer center. Methods: This is a retrospective cohort study of all surgical oncology and gynecologic oncology cases performed in the province of Ontario and at the University Health Network/Princess Margaret Cancer Center (UHN/PMH) as a surrogate for treatment delays, compared to all gynecologic oncology patients treated at the Dana Farber Cancer Institute, Boston, MA, USA, between March 3, 2020-June 30, 2020. Descriptive statistics and treatment times were analysed using SPSS 25.0 Results: In the province of Ontario, between March 15, 2020-October 25, 2020, there was a 19% overall decrease in surgical oncology volumes compared to the same time period the previous year (March 17, 2019-Oct 17, 2019). There was an increase of 96% in high priority surgical oncology cases and a 43% decrease in low priority cancer cases. Surgeries for gynecologic malignancies decreased by 8% in the province of Ontario as compared to the previous year. At UHN/PMH, a publicly funded tertiary cancer center in Ontario, there was a 59.8% reduction in surgical oncology volumes between March 09, 2020-May 04 2020, as compared to the previous year (March 04, 2019-April 29 2019). In comparison, at the Dana Farber Cancer Institute, a privately funded tertiary cancer center, there were 202 new gynecologic oncology patient referrals and 66 returning patients between March 3, 2020-June 30, 2020. The median time from referral to first consultation for new patients was 11 days (range 1-21) and the time to primary treatment was 31 days (range 5-157). New patients with ovarian malignancies had the shortest time to treatment of 22 days (range 5-157). There were no modifications made to the standard of care treatment plans for any patients and 5.7% of the patients had treatment delays. Conclusions: During the COVID-19 pandemic, the public Canadian healthcare system in the province of Ontario was subjected to a 19% reduction in surgical oncology volumes and 8% specifically for gynecologic oncology surgeries, while there was a 5.7% treatment delay for gynecologic oncology surgeries for a similar patient cohort treated at a tertiary cancer center in a privately funded health care system in the United States.

[This corrects the article DOI: 10.1021/acsptsci.0c00181.].

In this paper, dopamine (DA) ionic states in different pH solutions and corresponding electrochemical reaction characteristics on indium tin oxide (ITO) electrodes were discussed in detail. Polydopamine (PDA) and gold nanoparticles (AuNPs) have been used to modify ITO electrodes for selective sensing of DA in the present of ascorbic acid (AA). The impedance of PDA membranes and the catalysis of AuNPs on the interfacial properties of the modified electrodes were studied tentatively. Experiments show that a three-layer AuNP-PDA hybrid membrane-modified ITO electrode has an excellent property in DA detection without being interfered by the existence of AA. The sensitivity of DA detection was obtained to be 0.300 μA/μM with a detection limit of 0.06 μM (S/N = 3) in chronoamperometry measurement. The application of the electrode was verified in real sample analysis with both differential pulse voltammetry and chronoamperometry measurements.

Herein, we report a simple method of simultaneous detection of hydroquinone (HQ) and catechol (CC) by cyclic voltammetry (CV) using activated glassy carbon electrodes (GCE). It was found that the two isomers can be completely separated by CV method on cathode polarized GCE (PGCE) after its anodic oxidation. This facile processing method solves the difficulty of electrode surface regenerating which disturb most modified electrodes for the complex composition ease of being contaminated by the analytes in measurement. Morphology and composition of the PGCE were characterized by scanning electron microscope (SEM), Fourier transform infrared (FTIR) spectra, and X-ray photoelectron spectroscopy. In addition, the reaction kinetic of HQ and CC reaction on the PGCE was investigated. It was found that the reaction kinetics of HQ and CC is a surface adsorption-controlled process at low concentration and a diffusion-controlled process at high concentration.

Highly soluble and stable proteins are desirable for many different applications, from basic science to reaching a cancer patient in the form of a biological drug. For X-ray crystallography-where production of a protein crystal might take weeks and even months-a stable protein sample of high purity and concentration can greatly increase the chances of producing a well-diffracting crystal. For a patient receiving a specific protein drug, its safety, efficacy, and even cost are factors affected by its solubility and stability. Increased protein expression and protein stability can be achieved by randomly altering the coding sequence. As the number of mutants generated might be overwhelming, a powerful protein expression and stability screen is required. In this chapter, we describe a colony filtration technology, which allows us to screen random mutagenesis libraries for increased thermal stability-the Hot CoFi blot. We share how to create the random mutagenesis library, how to perform the Hot CoFi blot, and how to identify more thermally stable clones. We use the Tobacco Etch Virus protease as a target to exemplify the procedure.

Background: Long-acting injectable cabotegravir (CAB LA) is a novel integrase inhibitor in advanced clinical development for HIV treatment and HIV prevention. We investigated the terminal pharmacokinetics and safety of CAB LA after final injection as part of a Phase 2 study. Methods: We performed a randomized, double-blind placebo-controlled trial at 8 sites in the United States (US), sub-Saharan Africa, and Brazil. Participants, age 18-65, HIV-uninfected and at low-risk for HIV, were randomized 3:1 to CAB LA or placebo. Participants received CAB LA at one of two doses/intervals, which were aggregated for this analysis. The current analysis includes participants who received at least one injection and examines the tail phase (time from final injection to 52-76 weeks post-final injection). The outcomes were safety and pharmacokinetics. Safety was reported as incidence rates of grade 2 or higher adverse events. The apparent terminal phase half-life (t1/2app) and time to decay of CAB drug concentration to below the lower limit of quantification (LLOQ) were estimated using non-compartmental methods. Findings: Overall, 177 participants received at least one injection. Median age was 31 years, 66.1% were female, 69.5% were non-white, 52.5% were from the US. Rates of grade 2 or higher adverse events on CAB were higher in the injection phase compared to the tail phase. At one time point from 52-60 weeks and at 76 weeks after final injection, 22% and 13% of males, respectively, had detectable CAB concentrations; 63% and 48% of females had detectable CAB concentrations at those time points. The estimated median time from last injection to CAB concentration below the LLOQ was 43.7 (range 20-4-152.5) weeks for males and 67.3 (range 17.7-225.5) weeks for females. Terminal phase half-life was 33% longer for female sex at birth participants compared to male participants, and 31% longer for participants with body mass index (BMI) above vs. below the study median of 26. Interpretation: No new adverse events were observed with long-term follow-up. The long pharmacokinetic tail will need to be addressed in pivotal trials of treatment and prevention. Trial Registration: The trial was registered with ClinicalTrials.gov, number NCT02178800. This trial is completed. Funding Statement: National Institute of Allergy and Infectious Diseases (UM1AI068619, UM1AI068613, and, UM1AI068617) Declaration of Interests: RJL has received study medication, consulting fees and travel support from Gilead Sciences, and consulting fees and travel support from Merck, Inc. JJE is a consultant to Merck, Gilead Sciences, Janssen, and ViiV Healthcare; and he is an investigator on research contracts from ViiV Healthcare, Janssen, and Gilead Sciences. HD has received honoraria from Pfizer-South Africa, Novartis-South Africa, MSD-South Africa and Adcock Ingram for speaking engagements and has received travel support from Mylan-South Africa, Novartis-South Africa and Aspen-South Africa. AYL received research grants from NIH and has led studies in which study drug was donated by Gilead Sciences. MAM received grant support through the NIH and received grant support through ViiV/GSK on work external to this study. ARR and DM are paid employees of ViiV Healthcare. CWH has research contracts with NIH, ViiV/GlaxoSmithKline, Gilead Sciences, and Merck through Johns Hopkins. Ethics Approval Statement: Local institutional review boards or ethics committees at each participating site granted ethics and regulatory approvals of the protocol and all participants provided written informed consent.

Background Cabotegravir (CAB) is a novel strand-transfer integrase inhibitor being developed for HIV treatment and prevention. CAB is formulated both as an immediate-release oral tablet for daily administration and as a long-acting injectable suspension (long-acting CAB [CAB LA]) for intramuscular (IM) administration, which delivers prolonged plasma exposure to the drug after IM injection. HIV Prevention Trials Network study 077 (HPTN 077) evaluated the safety, tolerability, and pharmacokinetics of CAB LA in HIV-uninfected males and females at 8 sites in Brazil, Malawi, South Africa, and the United States. Methods and findings HPTN 077 was a double-blind, placebo-controlled phase 2a trial. Healthy individuals age 18–65 years at low HIV risk were randomized (3:1) to receive CAB or placebo (PBO). In the initial oral phase, participants received 1 daily oral tablet (CAB or PBO) for 4 weeks. Those without safety concerns in the oral phase continued and received injections in the injection phase (Cohort 1: 3 injections of CAB LA 800 mg or 0.9% saline as PBO IM every 12 weeks for 3 injection cycles; Cohort 2: CAB LA 600 mg or PBO IM for 5 injection cycles; the first 2 injections in Cohort 2 were separated by 4 weeks, the rest by 8 weeks). The primary analysis included weeks 5 to 41 of study participation, encompassing the injection phase. The cohorts were enrolled sequentially. Primary outcomes were safety and tolerability. Secondary outcomes included pharmacokinetics and events occurring during the oral and injection phases. Between February 9, 2015, and May 27, 2016, the study screened 443 individuals and enrolled 110 participants in Cohort 1 and 89 eligible participants in Cohort 2. Participant population characteristics were as follows: 66% female at birth; median age 31 years; 27% non-Hispanic white, 41% non-Hispanic black, 24% Hispanic/Latino, 3% Asian, and 6% mixed/other; and 6 transgender men and 1 transgender woman. Twenty-two (11%) participants discontinued the oral study product; 6 of these were for clinical or laboratory adverse events (AEs). Of those who received at least 1 CAB LA injection, 80% of Cohort 1 and 92% of Cohort 2 participants completed all injections; injection course completion rates were not different from those in the PBO arm. Injection site reactions (ISRs) were common (92% of Cohort 1 and 88% of Cohort 2 participants who received CAB LA reported any ISR). ISRs were mostly Grade 1 (mild) to Grade 2 (moderate), and 1 ISR event (Cohort 1) led to product discontinuation. Grade 2 or higher ISRs were the only AEs reported more commonly among CAB LA recipients than PBO recipients. Two Grade 3 (severe) ISRs occurred in CAB recipients, 1 in each cohort, but did not lead to product discontinuation in either case. Seven incident sexually transmitted infections were diagnosed in 6 participants. One HIV infection occurred in a participant 48 weeks after last injection of CAB LA: CAB was not detectable in plasma both at the time of first reactive HIV test and at the study visit 12 weeks prior to the first reactive test. Participants in Cohort 2 (unlike Cohort 1) consistently met prespecified pharmacokinetic targets of at least 95% of participants maintaining CAB trough concentrations above PA-IC90, and 80% maintaining trough concentrations above 4× PA-IC90. Study limitations include a modest sample size, a short course of injections, and a low-risk study population. Conclusions In this study, CAB LA was well tolerated at the doses and dosing intervals used. ISRs were common, but infrequently led to product discontinuation. CAB LA 600 mg every 8 weeks met pharmacokinetic targets for both male and female study participants. The safety and pharmacokinetic results observed support the further development of CAB LA, and efficacy studies of CAB LA for HIV treatment and prevention are in progress. Trial registration ClinicalTrials.gov Registry: ClinicalTrials.gov Trial number: NCT02178800., In a phase 2a trial, Raphael Landovitz and colleagues investigate the safety of injectable cabotegravir in HIV-uninfected people, Author summary Why was this study done? Daily oral pre-exposure prophylaxis (PrEP) with tenofovir disoproxil fumarate/emtricitabine is highly effective in preventing HIV infection when taken as prescribed, but adherence challenges have compromised full effectiveness in some populations. Cabotegravir (CAB) is a novel strand-transfer integrase inhibitor in development as a long-acting injectable preparation for HIV treatment and HIV prevention. HIV Prevention Trials Network study 077 (HPTN 077) aimed to establish the safety, tolerability, and pharmacokinetics of 2 dose/interval regimens of long-acting CAB in HIV-uninfected, low-risk individuals globally. What did the researchers do and find? Participants were randomized to active CAB or placebo. All participants received a 4-week daily oral lead-in of CAB or placebo, followed by a series of injections of CAB or placebo. Participants received either 800 mg as a split 2-ml injection every 12 weeks, or 600 mg as a single 3-ml injection every 8 weeks, after receiving 2 injections 4 weeks apart. We observed frequent injection site reactions (mostly injection site pain), and they were more common with CAB than placebo, but only led to discontinuation of injections in rare cases. No other safety concerns were noted. Pharmacokinetics, using target values from nonhuman primate simian/human immunodeficiency virus challenge prevention studies, support the 600 mg every 8 weeks dose, meeting prespecified targets. What do these findings mean? We found that CAB was generally well tolerated in HIV-uninfected males and females in diverse geographic locations. Injection site reactions, although frequent, did not deter continued dosing, albeit with short courses. Studies are ongoing to evaluate efficacy in at-risk populations and to further evaluate the safety of the 600 mg every 8 weeks dose.

AIM The local delivery of growth factors via gene therapy has gained tremendous awareness in recent years due to their sustained growth factor delivery to target tissues. The aim of this study was to fabricate and investigate a scaffold able to release growth factors via gene therapy for the repair of periodontal tissues. MATERIALS AND METHODS Novel mesoporous bioglass (MBG)/silk fibrin scaffold combined with BMP7 and/or PDGF-B adenovirus was fabricated and tested in vitro for cell migration, proliferation and differentiation. Furthermore, acute-type buccal dehiscence periodontal defects (mesiodistal width × depth: 5 × 5 mm) were created on the buccal portion of the maxillary premolars in five normal male beagle dogs (12 months old, 15.0 ± 2.0 kg) and histologically examined for periodontal regeneration following implantation of the following five groups: (1) no scaffold, (2) MBG/silk scaffold alone, (3) scaffold + adPDGF-B, (4) scaffold + adBMP7, (5) scaffold + adPDGF-b + adBMP7. RESULTS In vitro findings demonstrated that adPDGF-B was able to rapidly recruit periodontal ligament (PDL) cells over sixfold more effectively than adBMP7, whereas adBMP7 was more able to induce osteoblast differentiation of PDL cells. In vivo findings demonstrate that scaffolds loaded with adPDGF-B were able to partially regenerate the periodontal ligament while adBMP7 scaffolds primarily improved new bone formation. The combination of both adPDGF-B and adBMP7 synergistically promoted periodontal regeneration by allowing up to two times greater regeneration of the periodontal ligament, alveolar bone and cementum when compared to each adenovirus used alone. CONCLUSIONS Although both PDGF-B and BMP7 are individually capable of promoting periodontal regeneration to some degree, their combination synergistically promotes wound healing in acute-type buccal dehiscence periodontal defects when delivered simultaneously. This study demonstrates the promise for successful delivery of low-cost, effective growth factor delivery via gene therapy for the treatment of periodontal defects.

The viral tegument is a layer of proteins between the herpesvirus capsid and its outer envelope. According to phylogenetic studies, only a third of these proteins are conserved amongst the three subfamilies (Alpha-, Beta- and Gammaherpesvirinae) of the family Herpesviridae. Although some of these tegument proteins have been studied in more detail, the structure and function of the majority of them are still poorly characterized. VP22 from Herpes simplex virus 1 (subfamily Alphaherpesvirinae) is a highly interacting tegument protein that has been associated with tegument assembly. We have determined the crystal structure of the conserved core domain of VP22, which reveals an elongated dimer with several potential protein–protein interaction regions and a peptide-binding site. The structure provides us with the structural basics to understand the numerous functional mutagenesis studies of VP22 found in the literature. It also establishes an unexpected structural homology to the tegument protein ORF52 from Murid herpesvirus 68 (subfamily Gammaherpesvirinae). Homologues for both VP22 and ORF52 have been identified in their respective subfamilies. Although there is no obvious sequence overlap in the two subfamilies, this structural conservation provides compelling structural evidence for shared ancestry and functional conservation.