DEGAP: Dynamic elongation of a genome assembly path.

Genome assembly remains to be a major task in genomic research. Despite the development over the past decades of different assembly software programs and algorithms, it is still a great challenge to assemble a complete genome without any gaps. With the latest DNA circular consensus sequencing (CCS) technology, several assembly programs can now build a genome from raw sequencing data to contigs; however, some complex sequence regions remain as unresolved gaps. Here, we present a novel gap-filling software, DEGAP (Dynamic Elongation of a Genome Assembly Path), that resolves gap regions by utilizing the dual advantages of accuracy and length of high-fidelity (HiFi) reads. DEGAP identifies differences between reads and provides 'GapFiller' or 'CtgLinker' modes to eliminate or shorten gaps in genomes. DEGAP adopts an iterative elongation strategy that automatically and dynamically adjusts parameters according to three complexity factors affecting the genome to determine the optimal extension path. DEGAP has already been successfully applied to decipher complex genomic regions in several projects and may be widely employed to generate more gap-free genomes.

Heart failure management at home: a non-randomised prospective case-controlled trial (HeMan at Home).

BACKGROUND/OBJECTIVES: Heart failure (HF) is a growing clinical and economic burden for patients and health systems. The COVID-19 pandemic has led to avoidance and delay in care, resulting in increased morbidity and mortality among many patients with HF. The increasing burden of HF during the COVID-19 pandemic led us to evaluate the quality and safety of the Hospital at Home (HAH) for patients presenting to their community providers or emergency department (ED) with symptoms of acute on chronic HF (CHF) requiring admission. DESIGN/OUTCOMES: A non-randomised prospective case-controlled of patients enrolled in the HAH versus admission to the hospital (usual care, UC). Primary outcomes included length of stay (LOS), adverse events, discharge disposition and patient satisfaction. Secondary outcomes included 30-day readmission rates, 30-day ED usage and ED dwell time. RESULTS: Sixty patients met inclusion/exclusion criteria and were included in the study. Of the 60 patients, 40 were in the HAH and 20 were in the UC group. Primary outcomes demonstrated that HAH patients had slightly longer LOS (6.3 days vs 4.7 days); however, fewer adverse events (12.5% vs 35%) compared with the UC group. Those enrolled in the HAH programme were less likely to be discharged with postacute services (skilled nursing facility or home health). HAH was associated with increased patient satisfaction compared with Hospital Consumer Assessment of Healthcare Providers and Systems (HCAHPS) score in North Carolina. Secondary outcomes of 30-day readmission and ED usage were similar between HAH and UC. CONCLUSIONS: The HAH pilot programme was shown to be a safe and effective alternative to hospitalisation for the appropriately selected patient presenting with acute on CHF.

RNAi and CRISPR-Cas silencing E3-RING ubiquitin ligase AIP2 enhances soybean seed protein content.

The majority of plant protein in the world's food supply is derived from soybean (Glycine max). Soybean is a key protein source for global animal feed and is incorporated into plant-based foods for people, including meat alternatives. Soybean protein content is genetically variable and is usually inversely related to seed oil content. ABI3-interacting protein 2 (AIP2) is an E3-RING ubiquitin ligase that targets the seed-specific transcription factor ABI3. Silencing both soybean AIP2 genes (AIP2a and AIP2b) by RNAi enhanced seed protein content by up to seven percentage points, with no significant decrease in seed oil content. The protein content enhancement did not alter the composition of the seed storage proteins. Inactivation of either AIP2a or AIP2b by a CRISPR-Cas9-mediated mutation increased seed protein content, and this effect was greater when both genes were inactivated. Transactivation assays in transfected soybean hypocotyl protoplasts indicated that ABI3 changes the expression of glycinin, conglycinin, 2S albumin, and oleosin genes, indicating that AIP2 depletion increased seed protein content by regulating activity of the ABI3 transcription factor protein. These results provide an example of a gene-editing prototype directed to improve global food security and protein availability in soybean that may also be applicable to other protein-source crops.

Enzymatic degradation is an effective means to reduce aflatoxin contamination in maize.

BACKGROUND: Aflatoxins are carcinogenic compounds produced by certain species of Aspergillus fungi. The consumption of crops contaminated with this toxin cause serious detrimental health effects, including death, in both livestock and humans. As a consequence, both the detection and quantification of this toxin in food/feed items is tightly regulated with crops exceeding the allowed limits eliminated from food chains. Globally, this toxin causes massive agricultural and economic losses each year. RESULTS: In this paper we investigate the feasibility of using an aflatoxin-degrading enzyme strategy to reduce/eliminate aflatoxin loads in developing maize kernels. We used an endoplasmic reticulum (ER) targeted sub-cellular compartmentalization stabilizing strategy to accumulate an aflatoxin-degrading enzyme isolated from the edible Honey mushroom Armillariella tabescens and expressed it in embryo tissue in developing maize kernels. Three transgenic maize lines that were determined to be expressing the aflatoxin-degrading enzyme both at the RNA and protein level, were challenged with the aflatoxin-producing strain Aspergillus flavus AF13 and shown to accumulate non-detectable levels of aflatoxin at 14-days post-infection and significantly reduced levels of aflatoxin at 30-days post-infection compared to nontransgenic control Aspergillus-challenged samples. CONCLUSIONS: The expression of an aflatoxin-degrading enzyme in developing maize kernels was shown to be an effective means to control aflatoxin in maize in pre-harvest conditions. This aflatoxin-degradation strategy could play a significant role in the enhancement of both US and global food security and sustainability.

Metabolic engineering of soybean seeds for enhanced vitamin E tocochromanol content and effects on oil antioxidant properties in polyunsaturated fatty acid-rich germplasm.

Soybean seeds produce oil enriched in oxidatively unstable polyunsaturated fatty acids (PUFAs) and are also a potential biotechnological platform for synthesis of oils with nutritional omega-3 PUFAs. In this study, we engineered soybeans for seed-specific expression of a barley homogentisate geranylgeranyl transferase (HGGT) transgene alone and with a soybean γ-tocopherol methyltransferase (γ-TMT) transgene. Seeds for HGGT-expressing lines had 8- to 10-fold increases in total vitamin E tocochromanols, principally as tocotrienols, with little effect on seed oil or protein concentrations. Tocochromanols were primarily in δ- and γ-forms, which were shifted largely to α- and ß-tocochromanols with γ-TMT co-expression. We tested whether oxidative stability of conventional or PUFA-enhanced soybean oil could be improved by metabolic engineering for increased vitamin E antioxidants. Selected lines were crossed with a stearidonic acid (SDA, 18:4Δ6,9,12,15)-producing line, resulting in progeny with oil enriched in SDA and α- or γ-linoleic acid (ALA, 18:3Δ9,12,15 or GLA, 18:3Δ6,9,12), from transgene segregation. Oil extracted from HGGT-expressing lines had ≥6-fold increase in free radical scavenging activity compared to controls. However, the oxidative stability index of oil from vitamin E-enhanced lines was ~15% lower than that of oil from non-engineered seeds and nearly the same or modestly increased in oil from the GLA, ALA and SDA backgrounds relative to controls. These findings show that soybean is an effective platform for producing high levels of free-radical scavenging vitamin E antioxidants, but this trait may have negative effects on oxidative stability of conventional oil or only modest improvement of the oxidative stability of PUFA-enhanced oil.

Charges for Alcoholic Cirrhosis Exceed All Other Etiologies of Cirrhosis Combined: A National and State Inpatient Survey Analysis.

BACKGROUND: Inpatient charges for patients with cirrhosis are substantial. We aimed to examine trends in inpatient charges among patients with cirrhosis to determine the drivers of healthcare expenditures. We hypothesized that alcoholic cirrhosis (AC) was a significant contributor to overall expense. METHODS: We performed a retrospective analysis of the Health Care Utilization Project Nationwide Inpatient Sample Database 2002-2014 (annual cross-sectional data) and New York and Florida State Inpatient Databases 2010-2012 (longitudinal data). Adult patients with cirrhosis of the liver were categorized as AC versus all other etiologies of cirrhosis combined. Patient characteristics were analyzed using ordinary least squares regression modeling. A random effects model was used to evaluate 30-day readmissions. RESULTS: In total, 1,240,152 patients with cirrhosis were admitted between 2002 and 2014. Of these, 567,510 (45.8%) had a diagnosis of AC. Total charges for AC increased by 95.7% over the time period, accounting for 59.9% of all inpatient cirrhosis-related charges in 2014. Total aggregate charges for AC admissions were $28 billion and increased from $1.4B in 2002 to $2.8B by 2014. In the NIS and SID, patients with AC were younger, white and male. Readmission rates at 30, 60, and 90 days were all higher among AC patients. CONCLUSIONS: Inpatient charges for cirrhosis care are high and increasing. Alcohol-related liver disease accounts for more than half of these charges and is driven by sheer volume of admissions and readmissions of the same patients. Effective alcohol addictions therapy may be the most cost-effective way to substantially reduce inpatient cirrhosis care expenditures.

Impact of tetracycline-clay interactions on bacterial growth.

Antibiotics are extremely effective against bacterial infections due to their selective toxicity for bacteria rather than the host. Extensive use and misuse of antibiotics resulted in significant increases in antibiotic levels in aquatic and soil environments. Bacteria exposed to antibiotics with low concentrations may develop antibiotic resistance. In this study a swelling 2:1 clay mineral montmorillonite (MMT) and a non-swelling 1:1 clay mineral kaolinite were premixed with tetracycline (TC) of varying concentrations. The gram-negative bacteria Escherichia coli (E. coli) and Salmonella enterica (S. enterica) of both TC sensitive and TC resistant strains were tested for their growth in the presence TC-loaded clay minerals of different amounts and under different physico-chemical conditions. The antimicrobial activity of TC was significantly decreased in the presence of MMT. In the absence of MMT, no bacteria growth was found at a TC concentration 0.25mg/mL and above. On the contrast, in the presence of MMT, 50% growth was still found for a TC resistant E. coli at a TC concentration of 5mg/g. The influence of kaolinite was to a lesser degree. These results suggest that antimicrobial agents present in clayey soils could be responsible for possible mutation of bacteria of high antibiotic resistance.

Characterization and functional biology of the soybean aleurone layer.

BACKGROUND: Soybean is a globally important oil seed crop. Both the high protein and oil content of soybean seeds make this crop a lucrative commodity. As in higher eukaryotic species with available genomes, the functional annotation of most of soybean's genes still remains to be investigated. A major hurdle in the functional genomics of soybean is a rapid method to test gene constructs before embarking on stable transformation experiments. RESULTS: In this paper we describe the morphology and composition of the persistent single-cell aleurone layer that derives from the endosperm of developing soybean seeds. Its composition compared to cotyledonary tissue indicates the aleurone layer plays a role in both abiotic and biotic stress. The potential utility as the aleurone layer as a transient expression system in soybean was shown. As a near transparent single-cell layer it can be used as a transient expression system to study transgene expression and inter- and intra-cellular targeting as it is amenable to microscopic techniques. CONCLUSION: The transparent single cell aleurone layer was shown to be compositionally comparable to cotyledonary tissue in soybean with an enrichment in oxidative response proteins and shown to be a potential transient expression platform.

Marketing para comunicar servicios profesionales farmacéuticos: experiencia en una farmacia comunitaria de l'Alfàs del Pi / Marketing to comunicate pharmaceutical professional services: Experience of a community pharmacy in l'Alfàs del Pi

Objetivos: Iniciar un nuevo servicio remunerado para el control y cribado de factores de riesgo cardiovascular, promocionándolo mediante una novedosa campaña de marketing. Conocer el nivel de riesgo cardiovascular de nuestros usuarios, así como de algunos de sus principales factores modificables asociados. Material y métodos: Estudio observacional transversal en usuarios adultos que solicitaron participar en la campaña. Esta se denominó "First Date: una cita con tu corazón" (haciendo alusión todos sus materiales al conocido programa televisivo) y se ofreció a los pacientes un Menú Especial a 12€ compuesto por un Entrante (Perímetro abdominal, IMC, PA y Toma de pulso), Plato Principal (Colesterol total y Edad Vascular) y Postre (nivel RCV). Resultados: Realizamos 34 menús: edad media 57,8 años y 53% de mujeres. Encontramos valores por encima de los límites normales, en (% de la población total): perímetro abdominal (50%), IMC (18% sobrepeso y 5% obesidad), colesterol total (70,6%), VOP (23,5%) y PAS (14,7% PAS). Respecto a la PAD se encontraron diferencias significativas entre ambos sexos: 70.0 ± 2.0 mmHg en mujeres y 83.1 ± 3.2 mmHg en hombres (**p=0.0012 t test). También se encontraron diferencias entre sexos en el hábito tabáquico (el 75% de los fumadores eran hombres) y de consumo de alcohol (el 72,7% de consumidores habituales eran hombres). El 29,4% de la población tenían RCV medio y el 20,6% elevado. Encontramos pulso irregular sin diagnóstico previo en el 11,8% de pacientes (2 hombres y 2 mujeres). Se realizaron 4 derivaciones al médico, 7 derivaciones para control de PA en la farmacia, 26 recomendaciones sobre dieta y ejercicio, 6 recomendaciones sobre tabaco y 3 intervenciones en adherencia. Conclusiones: Ayudados por la campaña de marketing, se consiguió comunicar e iniciar adecuadamente nuestro servicio profesional (remunerado) de cribado y control de los factores de riesgo cardiovascular. Los valores encontrados en nuestra población tienen un margen de mejora elevado, sobre todo en cuanto a los niveles de PAD, colesterol total, perímetro abdominal y nivel de RCV

Objectives: Start a new paid service for the control and screening of cardiovascular risk factors, promoting it through a innovative marketing campaign. And know the level of cardiovascular risk of our users, as well as some of its main modifiable factors associated. Material and methods: Cross-sectional observational study, in adult users who requested to participate in the campaign. It was called "First Date: an appointment with your heart" (referring all its materials to the well-known television program) and patients were offered a 12€ Special Menu composed by a Starter (Abdominal perimeter, BMI, BP and pulse collection), Main Course (Total colesterol and Vascular Age) and Dessert (CV risk). Results: We made 34 menus: average age 57.8 years and 53% of them women. We found values above the normal limits, without significant differences between sexes in (% of the total population): abdominal perimeter (50%), BMI (18% overweight and 5% obesity), total cholesterol (70.6%), PWV (23.5%) and PAS (14.7% SBP). Regarding the DBP, significant differences were found between both sexes: 70.0 ± 2.0 mmHg in women and 83.1 ± 3.2 mmHg in men ( p = 0.0012 t test). Sex differences in smoking habits (75% of smokers were men) and alcohol consumption (72.7% of habitual users were men) were found. 29.4% of the population had mean CV risk and 20.6% elevated. We found an irregular pulse without previous diagnosis in 11.8% of patients (2 men and 2 women). There were 4 referrals to the doctor, 7 referrals for control of BP in the pharmacy, 26 recommendations about diet and exercise, 6 recommendations about tobacco and 3 interventions on adherence. Conclusions: Aided by the marketing campaign, we were able to communicate and properly initiate our Professional Service (paid) for Screening and Control of Cardiovascular Risk Factors. The values found in our population have a high margin of improvement, especially in terms of the levels of DBP, total cholesterol, abdominal perimeter and CV risk levels

Soybean-derived recombinant human epidermal growth factor protects against experimental necrotizing enterocolitis.

BACKGROUND: Epidermal Growth Factor (EGF) reduces necrotizing enterocolitis (NEC). However, its high cost virtually prohibits clinical use. To reduce cost, soybean expressing human EGF was developed. Here we report effectiveness of soybean-derived EGF in experimental NEC. METHODS: Newborn rats were subjected to the NEC-inducing regimen of formula feeding and hypoxia. Formula was supplemented with extract from EGF-expressing or empty soybeans. NEC pathology was determined microscopically. Localization of tight junction proteins JAM-A and ZO-1 was examined by immunofluorescence and levels of mucosal COX-2 and iNOS mRNAs by real time PCR. RESULTS: Soybean extract amounts corresponding to 150µg/kg/day EGF caused considerable mortality, whereas those corresponding to 75µg/kg/day EGF were well tolerated. There was no significant difference in NEC scores between animals fed plain formula and formula supplemented with empty soybean extract. Soybean-EGF-supplemented formula at 75µg/kg/day EGF significantly decreased NEC, attenuated dissociation of JAM-A and ZO-1 proteins from tight junctions, and reduced intestinal expression of COX-2 and iNOS mRNAs. CONCLUSION: Supplementation with soybean-expressed EGF significantly decreased NEC in the rat model. Soybean-expressed EGF may provide an economical solution for EGF administration and prophylaxis of clinical NEC.

Patient, Provider, and Practice Characteristics Associated with Inappropriate Antimicrobial Prescribing in Ambulatory Practices.

OBJECTIVE To reduce inappropriate antimicrobial prescribing across ambulatory care, understanding the patient-, provider-, and practice-level characteristics associated with antibiotic prescribing is essential. In this study, we aimed to elucidate factors associated with inappropriate antimicrobial prescribing across urgent care, family medicine, and pediatric and internal medicine ambulatory practices. DESIGN, SETTING, AND PARTICIPANTS Data for this retrospective cohort study were collected from outpatient visits for common upper respiratory conditions that should not require antibiotics. The cohort included 448,990 visits between January 2014 and May 2016. Carolinas HealthCare System urgent care, family medicine, internal medicine and pediatric practices were included across 898 providers and 246 practices. METHODS Prescribing rates were reported per 1,000 visits. Indications were defined using the International Classification of Disease, Ninth and Tenth Revisions, Clinical Modification (ICD-9/10-CM) criteria. In multivariable models, the risk of receiving an antibiotic prescription was reported with adjustment for practice, provider, and patient characteristics. RESULTS The overall prescribing rate in the study cohort was 407 per 1,000 visits (95% confidence interval [CI], 405-408). After adjustment, adult patients seen by an advanced practice practitioner were 15% more likely to receive an antimicrobial than those seen by a physician provider (incident risk ratio [IRR], 1.15; 95% CI, 1.03-1.29). In the pediatric sample, older providers were 4 times more likely to prescribe an antimicrobial than providers aged ≤30 years (IRR, 4.21; 95% CI, 2.96-5.97). CONCLUSIONS Our results suggest that patient, practice, and provider characteristics are associated with inappropriate antimicrobial prescribing. Future research should target antibiotic stewardship programs to specific patient and provider populations to reduce inappropriate prescribing compared to a "one size fits all" approach. Infect Control Hosp Epidemiol 2018;39:307-315.

Increasing Prevalence of Hepatitis C among Hospitalized Children Is Associated with an Increase in Substance Abuse.

OBJECTIVE: To evaluate the impact of substance abuse on pediatric hepatitis C virus (HCV) prevalence, we examined geographic and demographic data on inpatient hospitalizations in children with HCV. STUDY DESIGN: We examined hospitalizations in children using the Kids' Inpatient Database, a part of the Healthcare Cost and Utilization Project. We identified cases using the International Classification of Diseases, 9th edition, codes for HCV infection during 2006, 2009, and 2012. Nonparametric tests for trend were used to calculate trend statistics. RESULTS: From 2006 to 2012 nationally, the number of hospitalizations of children with HCV increased 37% (2.69 to 3.69 per 10 000 admissions; P < .001). The mean age of children hospitalized was 17.6 years (95% CI, 17.4-17.8). HCV cases among those 19-20 years of age represented 68% of the total HCV diagnoses, with a 54% increase over the years sampled (P < .001 for trend). The burden of HCV in children was highest in whites, those in the lowest income quartile, and in the Northeast and Southern regions of the US (all P < .0001). The prevalence of substance use among children with HCV increased from 25% in 2006 to 41% in 2012 (P < .001). CONCLUSION: The increases of HCV in hospitalized children are largely in teenagers, highly associated with substance abuse, and concentrated in Northeast and Southern states. These results strongly suggest that public health efforts to prevent and treat HCV will also need to include adolescents.

Predictors of Unplanned Hospitalization in Patients Receiving Outpatient Parenteral Antimicrobial Therapy Across a Large Integrated Healthcare Network.

BACKGROUND: Outpatient parenteral antimicrobial therapy (OPAT) prescribing has increased along with the trend toward early discharge of hospitalized patients who have infections. There is limited literature that assesses unplanned hospitalizations during OPAT. This study aims to elucidate the predictors of unplanned hospitalization in OPAT patients after discharge from acute-care facilities within Carolinas HealthCare System (CHS). Understanding these predictors may inform future interventions to improve treatment efficacy and patient outcomes. METHODS: The study cohort included hospitalized patients aged >19 years who initiated OPAT in an acute-care facility within CHS in 2014-2015. Patients who had OPAT prescribed at an ambulatory-care facility were excluded. The primary outcome was unplanned hospitalization anytime during the at-risk time from discharge through 90 days. RESULTS: The unplanned hospitalization rate for the cohort was 18.5%. In adjusted analysis, having OPAT delivered at a skilled nursing facility was associated with a 46% (incident risk ratio = 1.46; 95% confidence interval = 1.04-2.06) increased risk of an unplanned hospitalization compared with patients receiving OPAT at home after adjustment for demographics, comorbidities, indication, treatment duration, and antimicrobial prescribed. Infusion, dialysis, and rehabilitation centers had the lowest rates of unplanned hospitalizations. CONCLUSIONS: These results suggest that the location of OPAT delivery is associated with unplanned hospitalizations and that older patients need additional support during OPAT.

Aflatoxin-free transgenic maize using host-induced gene silencing.

Aflatoxins, toxic secondary metabolites produced by some Aspergillus species, are a universal agricultural economic problem and a critical health issue. Despite decades of control efforts, aflatoxin contamination is responsible for a global loss of millions of tons of crops each year. We show that host-induced gene silencing is an effective method for eliminating this toxin in transgenic maize. We transformed maize plants with a kernel-specific RNA interference (RNAi) gene cassette targeting the aflC gene, which encodes an enzyme in the Aspergillus aflatoxin biosynthetic pathway. After pathogen infection, aflatoxin could not be detected in kernels from these RNAi transgenic maize plants, while toxin loads reached thousands of parts per billion in nontransgenic control kernels. A comparison of transcripts in developing aflatoxin-free transgenic kernels with those from nontransgenic kernels showed no significant differences between these two groups. These results demonstrate that small interfering RNA molecules can be used to silence aflatoxin biosynthesis in maize, providing an attractive and precise engineering strategy that could also be extended to other crops to improve food security.

Proteome rebalancing in transgenic Camelina occurs within the enlarged proteome induced by ß-carotene accumulation and storage protein suppression.

Oilseed crops are global commodities for their oil and protein seed content. We have engineered the oilseed Camelina sativa to exhibit increased protein content with a slight decrease in oil content. The introduction of a phytoene synthase gene with an RNAi cassette directed to suppress the storage protein 2S albumin resulted in seeds with an 11-24 % elevation in overall protein. The phytoene synthase cassette alone produced enhanced ß-carotene content of an average 275 ± 6.10 µg/g dry seed and an overall altered seed composition of 11 % less protein and comparable nontransgenic amounts of both oil and carbohydrates. Stacking an RNAi to suppress the major 2S storage protein resulted in seeds that contain elevated protein and slight decrease in oil and carbohydrate amounts showing that Camelina rebalances its proteome within an enlarged protein content genotype. In both ß-carotene enhanced seeds with/without RNAi2S suppression, the seed size was noticeably enlarged compared to nontransgenic counterpart seeds. Metabolic analysis of maturing seeds indicate that the enhanced ß-carotene trait had the larger effect than the RNAi2S suppression on the seed metabolome. The use of a GRAS (generally regarded as safe) ß-carotene as a visual marker in a floral dip transformation system, such as Camelina, might eliminate the need for costly regulatory and controversial antibiotic resistance markers. ß-carotene enhanced RNAi2S suppressed Camelina seeds could be further developed as a rapid heterologous protein production platform in a nonfood crop leveraging its enlarged protein content and visual marker.

Effectiveness and Safety of Sofosbuvir-Based Regimens for Chronic HCV Genotype 3 Infection: Results of the HCV-TARGET Study.

BACKGROUND: Sofosbuvir (SOF) is active against all hepatitis C virus (HCV) genotypes, and SOF-based therapies lead to high rates of sustained virologic response (SVR). However, genotype 3 (GT3) HCV remains a challenge with lower SVR rates reported, particularly in patients with cirrhosis. This study reports the effectiveness and safety of SOF-based therapy in patients with GT3 HCV treated in clinical practice. METHODS: Hepatitis C Virus Therapeutic Registry and Research Network is an international, prospective observational study evaluating patients treated in usual clinical practice. Patients with GT3 HCV were analyzed to assess predictors of treatment response and adverse events using descriptive statistics and multivariable logistic regression. RESULTS: Treatment outcomes were available for 197 patients treated with SOF and ribavirin (RBV), with or without peginterferon, including 54% with cirrhosis and 49% who failed prior therapy. Of 178 patients treated with SOF/RBV, 60% achieved SVR at 12 weeks (SVR12), compared with 84% of 19 patients treated with SOF/peginterferon/RBV. For patients treated with SOF/RBV, the SVR12 rate was 58% in treatment-naive patients with cirrhosis, and 42% in those with cirrhosis who failed prior therapy. In noncirrhotic patients, SVR12 rates were 89% in treatment-naive and 88% in treatment-experienced patients. After controlling for age and sex, absence of cirrhosis (odds ratio [OR], 6.4; 95% confidence interval [CI], 2.78-14.74), albumin levels ≥3.2 g/dL (OR, 12.48; 95% CI, 3.86-40.33), and platelet count >10(5) cells/µL (OR, 7.44; 95% CI, 3.51-15.78) were associated with greater odds of SVR12 CONCLUSIONS: SVR rates were acceptable in patients with GT3 HCV without cirrhosis; however, in those with cirrhosis, treatment with SOF/RBV was suboptimal, highlighting the need for new therapies for this population.

Transgenic Soybean Production of Bioactive Human Epidermal Growth Factor (EGF).

Necrotizing enterocolitis (NEC) is a devastating condition of premature infants that results from the gut microbiome invading immature intestinal tissues. This results in a life-threatening disease that is frequently treated with the surgical removal of diseased and dead tissues. Epidermal growth factor (EGF), typically found in bodily fluids, such as amniotic fluid, salvia and mother's breast milk, is an intestinotrophic growth factor and may reduce the onset of NEC in premature infants. We have produced human EGF in soybean seeds to levels biologically relevant and demonstrated its comparable activity to commercially available EGF. Transgenic soybean seeds expressing a seed-specific codon optimized gene encoding of the human EGF protein with an added ER signal tag at the N' terminal were produced. Seven independent lines were grown to homozygous and found to accumulate a range of 6.7 +/- 3.1 to 129.0 +/- 36.7 µg EGF/g of dry soybean seed. Proteomic and immunoblot analysis indicates that the inserted EGF is the same as the human EGF protein. Phosphorylation and immunohistochemical assays on the EGF receptor in HeLa cells indicate the EGF protein produced in soybean seed is bioactive and comparable to commercially available human EGF. This work demonstrates the feasibility of using soybean seeds as a biofactory to produce therapeutic agents in a soymilk delivery platform.

Histochemical analysis of collagen fibers in giant cell fibroma and inflammatory fibrous hyperplasia.

OBJECTIVE: The aim was to investigate collagen fibers in giant cell fibroma, inflammatory fibrous hyperplasia, and oral normal mucosa. MATERIALS AND METHODS: Sixty-six cases were stained with picrosirius red. The slides were observed under polarization, followed by the measurement of the area and the percentage of the type I and type III collagens. The age and gender were obtained from the clinical records. RESULTS: No differences could be observed in both the area and percentage of the type I and type III collagens within the categories of lesions and normal mucosa. In the giant cells fibroma, a greater area and percentage of type I collagen could be identified in individuals of less than 41.5 years (p<0.05). CONCLUSION: The distribution of type I and type III collagen fibers in the studied lesions followed a similar pattern to that observed in the normal mucosa, indicating a normal collagen maturation process of type III to I. The study supports that multinucleated and stellate cells of the giant cell fibroma appear to be functional within collagen types III and I turnover. The greater amount of type I collagen identified in giant cell fibroma in individuals of less than 41.5 years reinforce the neoplastic nature of lesion.