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A major unmet need in the cystic fibrosis (CF) therapeutic landscape is the lack of effective treatments for nonsense CFTR mutations, which affect approximately 10% of CF patients. Correction of nonsense CFTR mutations via genomic editing represents a promising therapeutic approach. In this study, we tested whether prime editing, a novel CRISPR-based genomic editing method, can be a potential therapeutic modality to correct nonsense CFTR mutations. We generated iPSCs from a CF patient homozygous for the CFTR W1282X mutation. We demonstrated that prime editing corrected one mutant allele in iPSCs, which effectively restored CFTR function in iPSC-derived airway epithelial cells and organoids. We further demonstrated that prime editing may directly repair mutations in iPSC-derived airway epithelial cells when the prime editing machinery is efficiently delivered by helper-dependent adenovirus (HDAd). Together, our data demonstrated that prime editing may potentially be applied to correct CFTR mutations such as W1282X. [ABSTRACT FROM AUTHOR]

Water resources scarcity has threatened the coordinative development of demographics, society and economy. As a typical rapidly urbanizing area and an emerging megacity in China, Chengdu is confronting the pressure of inadequate water supply. The present study divides the macroeconomic factors that affect the water resource supply and demand balance into six major subsystems: water resources supply, water demand, water drainage, population, ecological environment and economy. The combining variable interaction description and predictive simulation models are applied to simulate the water supply and demand ratio (S:D) from 2005 to 2035. Further, this study designs different development scenarios to simulate the change of S:D ratios by altering the parameter values of driving factors. The results show that: (1) the S:D ratio will decline if the current development scenario continues, implying the serious water resources shortage and the severe water supply-demand conflict in Chengdu; (2) socio-economic water demand and wastewater/rainwater reuse are the key driving parameters of S:D ratio, especially the water consumption per ten thousand yuan of industrial value-added; (3) the S:D ratio will increase from 0.92 in the current baseline scenario to 1.06 in the integrated optimization scenario in 2025, and the long-term planning brings 2035 from 0.71 to 1.03, with the proportion of unconventional water supply rise to 38% and 61%, respectively. This study can provide a decision-making tool for policy-makers to explore plausible policy scenarios necessary for bridging the gap between the water supply and demand in megacities. [ABSTRACT FROM AUTHOR]

Based on the engineering practice of large cross-section highway tunnel, this paper reveals the space-time coordinated evolution law of the construction mechanical characteristics and deformation distribution of the support structure in the construction by half bench CD method through field test. At the same time, the mechanical response calculation model of the supporting structure in the partial excavation is constructed, and the mechanical characteristics of the support structure in the partial excavation process are analyzed by above mechanical calculation model. Then, the mechanical and deformation distribution of the feet-reinforcement bolt in the steel frame—foot-reinforcement bolt combined support system is analyzed under different levels of surrounding rock load and different structural parameters of the feet-reinforcement bolt. The research results show that: (1) The internal force of the supporting structure changes most obviously during the excavation of Part Ⅰ, Part Ⅱ and Part Ⅲ, and the internal force of the support structure gradually tends to be stable after a slight increase in the excavation of Part Ⅳ and Part Ⅴ; (2) The horizontal deformation and vertical deformation of the support structure mainly occur in the excavation process of Part Ⅰ, Part Ⅱ and Part Ⅲ, and the excavation of Part Ⅳ and Ⅴ has little effect on the deformation response of the structure. The vertical displacement of the supporting structure is larger than the horizontal displacement, and the dynamic response of the temporary diaphragm structure during tunnel excavation is shrinkage-expansion-shrinkage-expansion; (3) The bending strain of each measuring point decreases with the increase of the distance from the loading point, and the bending strain of section 1 and section 2 is much larger than that of the other three sections; (4) With the increase of the angle, the section position with strain close to 0 gradually moves to the deeper position of the bolt, and the axial strain of each section on the bolt gradually changes from positive strain to negative strain. [ABSTRACT FROM AUTHOR]

Background: The 2019 coronavirus disease (COVID-19) pandemic is a public health emergency of international concern and poses a challenge to the mental health and sleep quality of front-line medical staff (FMS). The aim of this study was to investigate the sleep quality of FMS during the COVID-19 outbreak in China and analyze the relationship between mental health and sleep quality of FMS. Methods: From February 24, 2020 to March 22, 2020, a cross-sectional study was performed with 543 FMS from a medical center in Western China. A self-reported questionnaire was used to collect data anonymously. The following tests were used: The Self-Rating Anxiety Scale (SAS) for symptoms of anxiety, the Beck Depression Inventory (BDI) for depressive symptoms, and the Pittsburgh Sleep Quality Index (PSQI) for sleep quality assessment. Results: Of the 543 FMS, 216 (39.8%) were classified as subjects with poor sleep quality. Anxiety (P<0.001), depression (P<0.001), and the prevalence of those divorced or widowed (P<0.05) were more common in FMS with poor sleep quality than in participants with good sleep quality. The FMS exhibiting co-occurrence of anxiety and depression were associated with worse scores on sleep quality than those medical staff in the other three groups/categories. The difference in sleep quality between the FMS with only depression and the FMS experiencing co-occurrence of anxiety and depression was statistically significant (P<0.05). However, there was no significant difference in sleep quality between the FMS experiencing only anxiety and the FMS with co-occurrence of anxiety and depression (P > 0.05). Conclusions: During the COVID-19 pandemic, there was a noteworthy increase in the prevalence of negative emotions and sentiments among the medical staff, along with poor overall sleep quality. We anticipate that this study can stimulate more research into the mental state of FMS during outbreaks and other public health emergencies. In addition, particular attention must be paid to enhance the sleep quality of FMS, along with better planning and support for FMS who are continuously exposed to the existing viral epidemic by virtue of the nature of their profession. [ABSTRACT FROM AUTHOR]

(E)-N,N-dimethyl-4-oxo-4-(4-(pyridin-4-yl)phenyl)but-2-enamide hydrochloride (IMB-YH-4py5-2H) is a novel Protein Kinase B (PknB) inhibitor with potent activity against Mycobacterium tuberculosis strains. In the present study, a sensitive and specific liquid chromatography/tandem mass spectrometry (LC-MS/MS) method was developed and validated to determine IMB-YH-4py5-2H in rat plasma. Sample pretreatment was achieved by liquid-liquid extraction with ethyl acetate, and separation was performed on an XTerra MS C18 column (2.1×50 mm, 3.5 μm) with gradient elution (methanol and 0.1% formic acid) at a flow rate of 0.3 mL/min. Detection was performed in multiple reaction monitoring (MRM) mode. Linear calibration curves were obtained over a concentration range of 1−100 ng/mL. The intra-day and inter-day precisions were lower than 8.46%, and the accuracies ranged from -8.71% to 12.36% at all quality control levels. The extraction recoveries were approximately 70%, and the matrix effects were negligible. All quality control samples were stable under different storage conditions. The validated method was successfully applied to a preclinical pharmacokinetic study in Sprague-Dawley rats. IMB-YH-4py5-2H demonstrated improved pharmacokinetic properties (higher exposure level) compared with its leading compound. IMB-YH-4py5-2H was also distributed throughout the lung pronouncedly, especially inside alveolar macrophages, indicating its effectiveness against lower respiratory infections. [ABSTRACT FROM AUTHOR]

CircRNAs are involved in a variety of human diseases, however, the expression profiles and the potential diagnostic value of circRNAs in human acute Stanford type A aortic dissection (AAAD) remains largely unknown. In this study, high-throughput RNA sequencing (RNA-Seq) was used to investigate the differentially expressed circRNAs, microRNAs (miRs) and mRNAs in human AAAD tissues (n = 10) compared with normal aortic tissues (n = 10). The results of RNA-Seq revealed that 506 circRNAs were significantly dysregulated (P<0.05, false discovery rate, FDR<0.05, fold change>2). The subsequent weighted gene correlation network analysis and the following co-expression network analysis revealed that tyrosine-protein kinase Fgr might play important roles in the occurrence and development of AAAD. According to the circRNA-miRNA-mRNA network, we found that the upstream regulatory molecule of Fgr is circMARK3. Finally, a receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of the serum circMARK3 as biomarkers for AAAD (cutoff value = 1.497, area under the curve = 0.9344, P < 0.0001, sensitivity = 90.0%, specificity = 86.7%). These results provided a preliminary landscape of circRNAs expression profiles and indicated that circMARK3 was a potential biomarker for AAAD diagnosis. [ABSTRACT FROM AUTHOR]

Pharmacological efficacy is based on the drug concentration in target tissues, which usually cannot be represented by the plasma concentration. The purpose of this study was to compare the pharmacokinetic characteristics of gemifloxacin in plasma and skeletal muscle and evaluate its tissue penetration in both healthy and MRSA (methicillin-resistant Staphylococcus aureus)-infected rats. A microdialysis (MD) combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated to determine free gemifloxacin concentrations in rat plasma and skeletal muscle simultaneously. The in vivo recoveries of MD were 23.21% ± 3.42% for skeletal muscle and 20.62% ± 3.19% for plasma, and were concentration independent. We provided evidence that the method developed here meets FDA requirements. Additionally, this method was successfully applied to the determination of free gemifloxacin in rats. Muscle and blood dialysates were collected after an 18 mg/kg intravenous bolus dose. The mean areas under the concentration-time curves (AUCs) from 0 to 9 h for skeletal muscle and plasma were 3641.50 ± 915.65 h*ng/mL and 7068.32 ± 1964.19 h*ng/mL in MRSA-infected rats and 3774.72 ± 700.36 h*ng/mL and 6927.49 ± 1714.86 h*ng/mL in healthy rats, respectively. There was no significant difference (P>0.05) in gemifloxacin exposure between healthy rats and MRSA-infected rats for plasma or muscle. The low ratio of AUC0-9 muscle to AUC0-9 plasma suggested lower drug exposure in skeletal muscle than in plasma for both healthy and MRSA-infected rats. Our study suggested that the administration of gemifloxacin according to drug levels in plasma to treat local infection is unreasonable and might result in an inadequate dose regimen. [ABSTRACT FROM AUTHOR]

Hypovascular pancreatic neuroendocrine tumors (hypo-PNETs) are often misdiagnosed as pancreatic ductal adenocarcinoma (PDAC). However, the treatment options and prognosis of PNETs and PDAC are substantially different. This retrospective study differentiated hypo-PNETs from PDAC using contrast-enhanced CT (CE-CT). Clinical data and CE-CT findings, including tumor location, size, boundary, pancreatic duct dilatation, local invasion or metastases, tumor contrast enhancement, and tumor-to-pancreas enhancement ratio, were compared between 39 PDACs and 18 hypo-PNETs. At CT imaging, hypo-PNETs showed a higher frequency of a well-defined margin and lower frequencies of pancreatic duct dilatation and local invasion or metastasis when compared with PDAC (p < 0.05 for all). The mean attenuation of hypo-PNETs at the arterial and portal venous phase was significantly higher than that of PDAC (p < 0.001, p = 0.003, respectively). Similar results were observed in tumor-to-pancreas enhancement ratio. Tumor attenuation and tumor-to-pancreas enhancement ratio at the arterial phase showed the largest area under the curve (AUC) of 0.888 and 0.812 with 83.3–88.9% of sensitivity and 61.6–77.0% of specificity. Pancreatic duct dilatation, local invasion or metastasis, and tumor attenuation at the portal venous phase also showed acceptable AUC (0.703–0.748). Thus CE-CT features, especially the enhancement degree at the arterial phases, may be useful for differentiating hypo-PNETs from PDAC using CE-CT. [ABSTRACT FROM AUTHOR]

A terrestrial laser scanner is a fast, high-precision data acquisition device, which has been applied more and more to the research area of forest inventory. In this study, a type of automated low-cost terrestrial laser scanner was designed and implemented based on a SICK LMS-511 two-dimensional laser scanning sensor and a stepper motor. The new scanner was named BEE(developed by the department of Electronic Engineering, Beijing Forestry University), which can scan the forest trees in three dimensions. The BEE scanner and its supporting software are specifically designed for forest inventory. The specific software was developed to smoothly control the BEE scanner and to acquire the data, including the angular data, range data, and intensity data, and the data acquired by the BEE scanner could be processed into point cloud data, a range map, and an intensity map. Based on the point cloud data, the trees were detected by a single slice of the single scan in a plot, and the local ground plane was fitted for each detected tree. Then the diameter at breast height (DBH), tree height, and tree position could be estimated automatically by using the specific software. The experiments have been performed by using the BEE scanner in an artificial ginkgo forest which was located in the Haidian District of Beijing. Four 10 m × 10 m square plots were selected for the experiments. The BEE scanner scanned in the four plots and acquired the single-scan data, respectively. The DBH, tree height, and tree position of the trees in the four plots were estimated and analyzed. For comparison, manually-measured data was also collected in the four plots. The trunk detection rate for all four plots was 92.75%; the root mean square error of the DBH estimation was 1.27 cm; the root mean square error of the tree height estimation was 0.24 m; and the tree position estimation was in line with the actual position. The scanner also was tested in more natural forest in the JiuFeng Forest Park. Two plots with a radius of 5 meters were scanned. Eleven trees in the plot with a flat ground were detected and DBH were estimated. But tree detection was failed in the other plot because of the undulating ground. Experimental results show that the BEE scanner can efficiently estimate the structure parameters of plantation trees and has good potential in practical applications of forest inventory. [ABSTRACT FROM AUTHOR]

A multivariate linear regression model was proposed to achieve short period prediction of PM2.5 (fine particles with an aerodynamic diameter of 2.5 μm or less). The main parameters for the proposed model included data on aerosol optical depth (AOD) obtained through remote sensing, meteorological factors from ground monitoring (wind velocity, temperature, and relative humidity), and other gaseous pollutants (SO2, NO2, CO, and O3). Beijing City was selected as a typical region for the case study. Data on the aforementioned variables for the city throughout 2015 were used to construct two regression models, which were discriminated by annual and seasonal data, respectively. The results indicated that the regression model based on annual data had (R2 = 0.766) goodness-of-fit and (R2 = 0.875) cross-validity. However, the regression models based on seasonal data for spring and winter were more effective, achieving 0.852 and 0.874 goodness-of-fit, respectively. Model uncertainties were also given, with the view of laying the foundation for further study. [ABSTRACT FROM AUTHOR]

A novel α-galactosidase of glycoside hydrolase family 36 was cloned from Bacillus coagulans, overexpressed in Escherichia coli, and characterized. The purified enzyme Aga-BC7050 was 85 kDa according to SDS-PAGE and 168 kDa according to gel filtration, indicating that its native structure is a dimer. With p-nitrophenyl-α-- galactopyranoside (pNPGal) as the substrate, optimal temperature and pH were 55 °C and 6.0, respectively. At 60 °C for 30 min, it retained > 50% of its activity. It was stable at pH 5.0–10.0, and showed remarkable resistance to proteinase K, subtilisin A, α-chymotrypsin, and trypsin. Its activity was not inhibited by glucose, sucrose, xylose, or fructose, but was slightly inhibited at galactose concentrations up to 100 mM. Aga-BC7050 was highly active toward pNPGal, melibiose, raffinose, and stachyose. It completely hydrolyzed melibiose, raffinose, and stachyose in < 30 min. These characteristics suggest that Aga-BC7050 could be used in feed and food industries and sugar processing. [ABSTRACT FROM AUTHOR]

The dynamics of tumor burden, secreted proteins or other biomarkers over time, is often used to evaluate the effectiveness of therapy and to predict outcomes for patients. Many methods have been proposed to investigate longitudinal trends to better characterize patients and to understand disease progression. However, most approaches assume a homogeneous patient population and a uniform response trajectory over time and across patients. Here, we present a mixture piecewise linear Bayesian hierarchical model, which takes into account both population heterogeneity and nonlinear relationships between biomarkers and time. Simulation results show that our method was able to classify subjects according to their patterns of treatment response with greater than 80% accuracy in the three scenarios tested. We then applied our model to a large randomized controlled phase III clinical trial of multiple myeloma patients. Analysis results suggest that the longitudinal tumor burden trajectories in multiple myeloma patients are heterogeneous and nonlinear, even among patients assigned to the same treatment cohort. In addition, between cohorts, there are distinct differences in terms of the regression parameters and the distributions among categories in the mixture. Those results imply that longitudinal data from clinical trials may harbor unobserved subgroups and nonlinear relationships; accounting for both may be important for analyzing longitudinal data. [ABSTRACT FROM AUTHOR]

Accurate and efficient detection of sleep spindles is a methodological challenge. The present study describes a method of using non-experts for manual detection of sleep spindles. We recruited five experts and 168 non-experts to manually identify spindles in stage N2 and stage N3 sleep data using a MATLAB interface. Scorers classified each spindle into definite and indefinite spindle (with weights of 1 and 0.5, respectively). First, a method of optimizing the thresholds of the expert/non-expert group consensus according to the results of experts and non-experts themselves is described. Using this method, we established expert and non-expert group standards from expert and non-expert scorers, respectively, and evaluated the performance of the non-expert group standards by compared with the expert group standard (termed EGS). The results indicated that the highest performance was the non-expert group standard when definite spindles were only considered (termed nEGS-1; F1 score = 0.78 for N2; 0.68 for N3). Second, four automatic spindle detection methods were compared with the EGS. We found that the performance of nEGS-1 versus EGS was higher than that of the four automated methods. Our results also showed positive correlation between the mean F1 score of individual expert in EGS and the F1 score of nEGS-1 versus EGS across 30 segments of stage N2 data (r = 0.61, P < 0.001). Further, we found that six and nine non-experts were needed to manually identify spindles in stages N2 and N3, respectively, while maintaining acceptable performance of nEGS-1 versus EGS (F1 score = 0.79 for N2; 0.64 for N3). In conclusion, this study establishes a detailed process for detection of sleep spindles by non-experts in a crowdsourcing scheme. [ABSTRACT FROM AUTHOR]

The literature on team cooperation has neglected the effects of relative kindness intention on cooperation, which we measure by comparing the kindness intentions of an agent to her group members to the kindness shown by other members to this same agent. We argue that the agent’s emotional reaction to material payoff inequity is not constant, but rather affected by her relative kindness intention. Then, we apply the model to team projects with multiple partners and investigate how inequity-aversion and relative kindness intention jointly influence team cooperation. We first consider the case of homogeneous agents, where their marginal productivity levels and technical capacities are the same, and then consider the case of heterogeneous agents, where their marginal productivity levels and technical capacities are not the same. Our results show that inequity-aversion has no effect on effort expenditure in the former case, but does affect it in the latter case. The consideration of relative kindness intention may impact the agents’ optimal cooperative effort expenditure when their technical capacities are different. In addition, it is beneficial for team cooperation, and might not only reduce the negative impact but also enhance the positive impact of inequity-aversion on the agents’ effort expenditures. [ABSTRACT FROM AUTHOR]

Phishing is the spoofing of Internet websites or emails aimed at tricking users into entering sensitive information, with such goals as financial or identity theft. The current study sought to determine whether age is associated with increased susceptibility to phishing and whether tests of executive functioning can predict phishing susceptibility. A total of 193 cognitively intact participants, 91 younger adults and 102 older adults, were primarily recruited through a Psychology department undergraduate subject pool and a gerontology research registry, respectively. The Executive Functions Module from the Neuropsychological Assessment Battery and the Iowa Gambling Task were the primary cognitive predictors of reported phishing suspiciousness. Other predictors included age group (older vs. younger), sex, education, race, ethnicity, prior knowledge of phishing, prior susceptibility to phishing, and whether or not browsing behaviors were reportedly different in the laboratory setting versus at home. A logistic regression, which accounted for a 22.7% reduction in error variance compared to the null model and predicted phishing suspiciousness with 73.1% (95% CI [66.0, 80.3]) accuracy, revealed three statistically significant predictors: the main effect of education (b = 0.58, SE = 0.27) and the interactions of age group with prior awareness of phishing (b = 2.31, SE = 1.12) and performance on the Neuropsychological Assessment Battery Mazes test (b = 0.16, SE = 0.07). Whether or not older adults reported being suspicious of the phishing attacks used in this study was partially explained by educational history and prior phishing knowledge. This suggests that simple educational interventions may be effective in reducing phishing vulnerability. Although one test of executive functioning was found useful for identifying those at risk of phishing susceptibility, four tests were not found to be useful; these results speak to the need for more ecologically valid tools in clinical neuropsychology. [ABSTRACT FROM AUTHOR]

It has become increasingly clear that proper cellular control of pluripotency and differentiation is related to the regulation of rRNA synthesis. To further our understanding of the role that the regulation of rRNA synthesis has in pluripotency we monitored rRNA synthesis during the directed differentiation of human embryonic stem cells (hESCs). We discovered that the rRNA synthesis rate is reduced ~50% within 6 hours of ACTIVIN A treatment. This precedes reductions in expression of specific stem cell markers and increases in expression of specific germ layer markers. The reduction in rRNA synthesis is concomitant with dissociation of the Pol I transcription factor, UBTF, from the rRNA gene promoter and precedes any increase to heterochromatin throughout the rRNA gene. To directly investigate the role of rRNA synthesis in pluripotency, hESCs were treated with the Pol I inhibitor, CX-5461. The direct reduction of rRNA synthesis by CX-5461 induces the expression of markers for all three germ layers, reduces the expression of pluripotency markers, and is overall similar to the ACTIVIN A induced changes. This work indicates that the dissociation of UBTF from the rRNA gene, and corresponding reduction in transcription, represent early regulatory events during the directed differentiation of pluripotent stem cells. [ABSTRACT FROM AUTHOR]

The α-hemolysin, encoded by the hla gene, is a major virulence factor in S. aureus infections. Changes in key amino acid residues of α-hemolysin can result in reduction, or even loss, of toxicity. The aim of this study was to investigate the diversity of the hla gene sequence and the relationship of hla variants to the clonal background of S. aureus isolates. A total of 47 clinical isolates from China were used in this study, supplemented with in silico analysis of 318 well-characterized whole genome sequences from globally distributed isolates. A total of 28 hla genotypes were found, including three unique to isolates from China, 20 found only in the global genomes and five found in both. The hla genotype generally correlated with the clonal background, particularly the multilocus sequence type, but was not related to geographic origin, host source or methicillin-resistance phenotype. In addition, the hla gene showed greater diversity than the seven loci utilized in the MLST scheme for S. aureus. Our investigation has provided genetic data which may be useful for future studies of toxicity, immunogenicity and vaccine development. [ABSTRACT FROM AUTHOR]

Upland cotton (Gossypium hirsutum L., 2n = 52, AADD) is an allotetraploid, therefore the discovery of single nucleotide polymorphism (SNP) markers is difficult. The recent emergence of genome complexity reduction technologies based on the next-generation sequencing (NGS) platform has greatly expedited SNP discovery in crops with highly repetitive and complex genomes. Here we applied restriction-site associated DNA (RAD) sequencing technology for de novo SNP discovery in allotetraploid cotton. We identified 21,109 SNPs between the two parents and used these for genotyping of 161 recombinant inbred lines (RILs). Finally, a high dense linkage map comprising 4,153 loci over 3500-cM was developed based on the previous result. Using this map quantitative trait locus (QTLs) conferring fiber strength and Verticillium Wilt (VW) resistance were mapped to a more accurate region in comparison to the 1576-cM interval determined using the simple sequence repeat (SSR) genetic map. This suggests that the newly constructed map has more power and resolution than the previous SSR map. It will pave the way for the rapid identification of the marker-assisted selection in cotton breeding and cloning of QTL of interest traits. [ABSTRACT FROM AUTHOR]

Background: Commercial sex is one of the major modes of HIV transmission in China. Understanding HIV risk behaviours in female sex workers (FSW) is of great importance for prevention. This study aims to assess the magnitude and temporal changes of risk behaviours in Chinese FSW. Method: Five electronic databases were searched to identify peer-reviewed English and Chinese language articles published between January 2000 and December 2012 that reported risk behaviours among FSW in China, including condom use, HIV testing, and drug use. Linear regression and Spearman's rank correlation were used to examine temporal trends in these risk factors. The study followed PRISMA guidelines for meta-analyses and was registered in the PROSPERO database for systematic reviews. Results: A total of 583 articles (44 English, 539 Chinese) investigating 594,583 Chinese FSW were included in this review. At last sex, condom use was highest with commercial partners (clients), increasing from 53.7% in 2000 to 84.9% in 2011. During this same time period, condom use increased with regular partners from 15.2% to 40.4% and with unspecified partners from 38.6% to 82.5%. Increasing trends were also found in the proportion of sampled FSW who reported testing for HIV in the past 12 months (from 3.2% in 2000 to 48.0% in 2011), while drug use behaviours decreased significantly from 10.9% to 2.6%. Conclusion: During the first decade of 2000, Chinese FSWs’ self-reported risk behaviours have decreased significantly while HIV testing has increased. Further outreach and intervention efforts are needed to encourage condom use with regular partners, continue promotion of HIV testing, and provide resources for the most vulnerable FSW, particularly low tier FSW, who may have limited access to sexual health and prevention programs. [ABSTRACT FROM AUTHOR]