High end-of-treatment hepatitis B core-related antigen levels predict hepatitis flare after stopping nucleot(s)ide analogue therapy.

BACKGROUND AND AIMS: Accurate biomarkers to predict outcomes following discontinuation of nucleos(t)ide analogue (NA) therapy are needed. We evaluated serum hepatitis B core-related antigen (HBcrAg) level as a biomarker for predicting outcomes after NA discontinuation. METHODS: Patients with HBeAg-negative chronic hepatitis B (CHB) without cirrhosis were enrolled in a prospective trial evaluating clinical outcomes until 96 weeks after NA discontinuation. End of treatment (EOT) and off-treatment levels of serum HBcrAg, HBsAg, HBV RNA and HBV DNA were used to predict key clinical outcomes including hepatitis flare (ALT ≥5 × ULN and HBV DNA > 2000 IU/mL). The SCALE-B score was calculated for the purposes of model validation. RESULTS: HBcrAg was tested amongst 65 participants. The median age was 54 years, 54% were male and 83% were Asian. HBcrAg was detectable in 86% patients. HBcrAg level ≥4 log U/mL at EOT was predictive of hepatitis flare [8/10 (80%) vs. 17/55 (31%), p = .001]. The presence of either HBcrAg ≥4 log U/mL or detectable HBV RNA at EOT predicted for both biochemical relapse and hepatitis flare. The SCALE-B model at EOT predicted for virological relapse, biochemical relapse, hepatitis flare and HBsAg loss in this cohort. An increase in the serum HBcrAg level off-treatment was also associated with hepatitis flare. No participant with EOT HBcrAg level ≥4 log U/mL achieved HBsAg loss. CONCLUSIONS: High levels of serum HBcrAg predict for hepatitis flare after stopping NA therapy and low likelihood of HBsAg loss at week 96. People with high levels of serum HBcrAg are not suitable candidates for NA discontinuation.

Hepatitis B Virus Flares After Nucleot(s)ide Analogue Cessation Are Associated With Activation of Toll-Like Receptor Signaling Pathways.

BACKGROUND: We evaluated the patterns of peripheral Toll-like receptor (TLR) signaling activity and the expression of TLRs and natural killer (NK) cell activation in a cohort of patients experiencing severe hepatitis flares after stopping nucleot(s)ide analogues (NAs) therapy. METHODS: Samples were collected longitudinally from patients with chronic hepatitis B who were enrolled in a prospective study of NA discontinuation. Patients experiencing hepatitis flares were compared with patients with normal alanine aminotransferase. Peripheral blood mononuclear cells (PBMCs) were stimulated with TLR ligands and cytokine secretion in the cell culture supernatant measured. Expression of TLR2/4, NKG2D, NKp46, and triggering receptor expressed on myeloid cells 1 (TREM-1) on monocytes, NK, and NK-T cells was measured. RESULTS: Seventeen patients with severe reactivation hepatitis flares were compared to 12 nonflare patients. Hepatitis flares were associated with increased activity of TLR2-8 and TLR9 signaling in PBMCs at the time of peak flare compared to baseline. Hepatitis flares were also associated with (1) upregulation of TLR2 and (2) TREM-1 receptor expression on NK. There were no differences at baseline between flare patients and nonflare patients. CONCLUSIONS: Hepatitis flares off NA therapy have a significant innate inflammatory response with upregulation of TLR signaling on peripheral monocytes and TLR2 and TREM-1 expression on NK cells. This implicates the innate immune system in the immunopathogenesis of hepatitis B flares.

Proof of Concept for Shoot Blight and Fire Blight Canker Management with Postinfection Spray Applications of Prohexadione-Calcium and Acibenzolar-S-Methyl in Apple.

To reduce the severity of shoot blight and prevent the resulting development of cankers on perennial apple wood, we evaluated eight fire blight postinfection spray programs of prohexadione-calcium (PCA) alone or with acibenzolar-S-methyl (ASM) over 2 years. On mature trees of cultivar Royal Court, a single application of the high PCA rate (247 mg/liter) at 2 to 3 days after inoculation resulted in 89.5 and 69.5% reduction of shoot blight severity after inoculation. Two applications of PCA 247 mg/liter 12 or 14 days apart, with the first one applied 2 to 3 days after inoculation, resulted in 78.8 and 74.5% reduction of shoot blight severity in both years. A 100% control of canker incidence on perennial wood from infected shoots in both years was achieved with a single application of PCA (247 mg/liter) applied at 2 or 3 days after the inoculation, and three applications of PCA (125 mg/liter) + ASM (25 mg/liter) 12 to 16 days apart reduced canker incidence by 83.5 and 69% in the 2 years. The other programs with lower PCA rates and frequencies of application reduced shoot blight severity 50.8 and 51.8% (PCA) and 62.6 to 72% and 59.3% (PCA + ASM) over 2 years, respectively. Reduction of canker incidence on wood by the other programs was 66.5% and 69 to 90.4% in the two years, respectively. As fire blight cankers lead to death of dwarf apple trees and serve as primary sources of inoculum, our effective PCA and PCA + ASM programs could serve as viable postinfection management options. These treatments can reduce or prevent canker development and thus significantly abate tree losses in high-density apple orchards after fire blight epidemics occur.

Baseline serum HBV RNA is associated with the risk of hepatitis flare after stopping nucleoside analog therapy in HBeAg-negative participants.

BACKGROUND AND AIMS: HBV RNA in peripheral blood reflects HBV cccDNA transcriptional activity and may predict clinical outcomes. The prospective Melbourne HBV-STOP trial studied nucleot(s)ide analog discontinuation in HBeAg-negative non-cirrhotic participants with long-term virological suppression. Ninety-six weeks after stopping treatment, the proportion of participants with virological relapse (HBV DNA > 2000 IU/mL), biochemical relapse (ALT > 2 × ULN and HBV DNA > 2000 IU/mL), or hepatitis flare (ALT > 5 × ULN and HBV DNA > 2000 IU/mL) was 89%, 58%, and 38%, respectively. We evaluated the ability of serum HBV RNA levels to predict these outcomes. APPROACH RESULTS: HBV RNA levels were measured using the Roche cobas 6800/8800 HBV RNA Investigational Assay. Sixty-five participants had baseline and longitudinal off-treatment specimens available for RNA testing. HBV RNA was detectable at baseline in 25% of participants and was associated with a higher risk of biochemical relapse (81% vs. 51%, p value 0.04) and hepatitis flare (63% vs. 31%, p value 0.04). Participants who had undetectable serum HBV RNA as well as HBsAg ≤ 100 IU/mL at baseline were less likely to experience virological relapse (4 of 9, 44%) than participants with detectable HBV RNA and HBsAg level > 100 IU/mL (15/15, 100%; p value 0.0009). Off-treatment levels of HBV RNA were correlated with HBV DNA and were associated with the risk of hepatitis flare. CONCLUSIONS: Serum HBV RNA may be a useful biomarker for guiding clinical decision-making before stopping nucleot(s)ide analog therapy. Baseline HBV RNA and HBsAg levels are associated with the risk of clinical relapse, hepatitis flare, and disease remission off-treatment.

Fire blight resistance, irrigation and conducive wet weather improve Erwinia amylovora winter survival in cankers.

Erwinia amylovora causes fire blight, a disease responsible for enormous economic losses in the pome fruit-producing areas where it is present. Despite the abundant research on fire blight, information about E. amylovora population dynamics and survival in fire blight cankers and the plant defense responses to this pathogen in the infected bark are limited. In our study, we obtained fire blight cankers in apple, pear, and Asian pear cultivars showing differing resistance to the disease by shoot inoculation with E. amylovora. We collected cankers from irrigated and non-irrigated trees every 3 months in two independent field experiments and analyzed samples by viability digital PCR. We also assessed the expression of pathogenicity-related (PR) genes in the bark of selected apple and Asian pear cultivars. A logistic regression analysis revealed the impact of environmental and host factors on E. amylovora detection rates in cankers. The chances of detecting live E. amylovora cells in cankers increased significantly in those collected from irrigated trees, in July, and/or during an experiment performed in a year with an expected average rainfall when compared to samples from non-irrigated trees, collected in January, and/or during an experiment performed under environmental conditions dominated by drought. We found a positive correlation between the pathogen detection rates in cankers and the host resistance to fire blight that might be explained by lower E. amylovora survival rates in more damaged tissues of susceptible hosts. The genes PR-1, PR-2, PR-5, and PR-8 were induced in the bark surrounding apple and Asian pear fire blight cankers. Our study, involving the analysis of more than 800 canker samples, provides new knowledge about the fire blight disease cycle and lays the foundation for improved fire blight management and eradication strategies in pome fruit orchards.

Stopping nucleot(s)ide analogues in non-cirrhotic HBeAg-negative chronic hepatitis B patients: HBsAg loss at 96 weeks is associated with low baseline HBsAg levels.

BACKGROUND AND AIMS: Current guidelines recommend long-term nucleot(s)ide analogue (NA) therapy for patients with HBeAg-negative chronic hepatitis B (CHB). However, disease remission has been described after stopping NA therapy, as well as HBsAg loss. METHODS: We performed a prospective multi-centre cohort study of stopping NA therapy. Inclusion criteria were HBeAg-negative CHB, the absence of cirrhosis and HBVDNA5× ULN occurred in 35 (32%); ALT flares were not associated with HBsAg loss. There were no unexpected safety issues. CONCLUSION: Virological reactivation was very common after stopping NA therapy and occurred earlier after stopping TDF versus ETV. The majority of patients had ALT <2× ULN at week 96, but only one-third achieved disease remission and HBsAg loss was rare. Very low HBsAg levels at baseline were uncommon but predicted for HBsAg loss and disease remission.

Quasi-static and dynamic investigation of an advanced high strength and corrosion resistant 10 % Cr nanocomposite martensitic steel.

The mechanical response of an advanced high strength and corrosion resistant 10 % Cr nanocomposite steel (ASTM A1035CS Grade 120) is measured under uniaxial tension and compression at the strain rates of 10-4 s-1, 10-2 s-1, 100 s-1, 700 s-1, and 3000 s-1. The experiments are performed at 22 °C as well as 80 °C to investigate the material behavior at the expected temperature rise due to adiabatic deformation at 15 % strain. Additionally, different compression-shear hat-shaped specimens are tested at quasi-static and dynamic strain rates to investigate the localization behavior of this material. The material exhibits small strain rate sensitivity (SRS) during quasi-static loading, but a pronounced SRS between quasi-static and dynamic strain rates. Tension-compression asymmetry is also observed at both temperatures. Experiments at 80 °C reveal a decrease in flow stress in both tension and compression indicating the material is sensitive to thermal softening due to adiabatic heating. Load-Unload-Reload (LUR) and strain rate jump experiments are performed to investigate the reasoning behind the approximate rate insensitivity of ASTM A1035CS steel during quasi-static strain rates. A new constitutive model is also developed using a novel rate dependent material model with a modified Hockett-Sherby (MHS) hardening model and incorporating Lode angle dependence to capture the tension-compression asymmetry. The model is also used to predict the LUR and strain rate jump experiments. Finally, reasoning behind the unique rate dependent thermo-mechanical behavior of ASTM A1035CS steel is discussed in regards to adiabatic heating, strain-partitioning, and phase transformation.

In vivo confocal endomicroscopy in the diagnosis and evaluation of celiac disease.

BACKGROUND & AIMS: Accurate histopathology of endoscopic duodenal biopsy specimens is critical in the diagnosis of celiac disease (CD) but sampling error and poor quality specimens may generate a false-negative result. Confocal endomicroscopy (CEM) is a novel technology allowing real-time in vivo microscopy of the mucosa that may diagnose CD and evaluate its severity and response to treatment more accurately than histopathology. METHODS: Subjects with CD and controls prospectively underwent CEM. Features of villous atrophy and crypt hypertrophy were defined. A CEM score measuring CD severity was devised and validated against the diagnosis of CD and blinded histopathology. Receiver operator characteristics, sensitivity to change after treatment, and reliability of findings were assessed. RESULTS: From 31 patients (6 untreated CD, 11 treated CD, and 14 controls), 7019 CEM images paired with 326 biopsy specimens were obtained. The accuracy of CEM in diagnosing CD was excellent (receiver operator characteristics area under the curve, 0.946; sensitivity, 94%, specificity, 92%) and correlated well with the Marsh grading (R-squared, 0.756). CEM differentiated CD from controls (P < .0001) and was sensitive to change after treatment with gluten-free diet (1787 optical biopsies; P = .012). The intraclass correlation of reliability was high (0.759-0.916). Of the 17 cases with diagnosed CD, 16 (94%) were diagnosed correctly using CEM but only 13 (76%) had detectable histopathology changes. The procedure was safe and well-tolerated. CONCLUSIONS: CEM effectively diagnoses and evaluates CD severity in vivo. This promising technique has the potential to improve endoscopy efficiency.

Publisher Correction: Development of a viability digital PCR protocol for the selective detection and quantification of live Erwinia amylovora cells in cankers.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Development of a viability digital PCR protocol for the selective detection and quantification of live Erwinia amylovora cells in cankers.

Fire blight is a devastating disease of apple and pear caused by the bacterium Erwinia amylovora. One of its main symptoms is canker formation on perennial tissues which may lead to the death of limbs and/or the entire tree. E. amylovora overwinters in cankers which play an important role in initiating fire blight epidemics. However, knowledge of pathogen biology in cankers is scarce, in part due to limitations of classical microbiology methods and the inability of most molecular techniques to distinguish live from dead cells. In this work, a viability digital PCR (v-dPCR) protocol using propidium monoazide (PMA) was developed, allowing for the first time the selective detection and absolute quantification of E. amylovora live cells in apple and pear cankers collected in two time periods. Some key factors affecting the v-dPCR performance were the maceration buffer composition, the target DNA amplicon length, the thermal cycle number and the use of sodium dodecyl sulfate or PMA enhancer for Gram-negative bacteria to improve the effect of PMA. In the future, this methodology could shed light on E. amylovora population dynamics in cankers and provide clues on the effect of management practices, host cultivar, host water/nutritional status, etc., on bacterial survival.

Clinically significant small-bowel pathology identified by double-balloon enteroscopy but missed by capsule endoscopy.

BACKGROUND: Capsule endoscopy (CE) is increasingly being used to investigate the small bowel for various indications, including obscure GI bleeding (OGB). However, false negatives have been described. Double-balloon enteroscopy (DBE) is a new endoscopic technique developed to potentially view the entire small intestine while allowing therapeutic options to be carried out when appropriate. OBJECTIVE: We described 4 patients with small-bowel pathology missed on CE but detected by DBE. DESIGN: Descriptive retrospective study. All patients underwent CE followed by DBE. SETTING: Single-center tertiary referral hospital. PATIENTS: Four patients were included. Three patients had OGB that required blood transfusions. One patient with celiac disease, compliant on a strict gluten-free diet for 5 months, presented with persistent weight loss and abdominal pain. INTERVENTIONS: DBE followed by surgical exploration and resection of small-bowel pathology. MAIN OUTCOME MEASUREMENTS: Successful identification of pathology missed by CE. Definitive treatment of small-bowel pathology by surgical resection. RESULTS: CE did not identify the small-bowel pathology in all 4 patients. The 3 patients with OGB had small-bowel masses found by DBE. Two of these were GI stromal tumors and one was an adenocarcinoma. The patient with celiac disease had a malignant ulcer, confirmed to be a lymphoma after surgical resection. LIMITATIONS: Retrospective study and small sample size. CONCLUSIONS: CE and DBE are complementary investigations. If there is a high index of suspicion of small-bowel pathology despite a negative CE, DBE should be performed.

Chondromyxoid fibroma of the upper thoracic spine in a 7-year-old patient. A case report and review of the literature.

BACKGROUND: We report a case of a 7-year-old white female who presented with acute, progressive bilateral lower extremity weakness over 48 h. METHODS: Case report and presentation of clinical, radiological and pathological data on a single case of chondromyxoid fibroma (CMF) of the T2 vertebral body. RESULTS: Magnetic resonance imaging of the thoracic spine revealed an extensive mass invading the lamina of the second thoracic vertebra, causing extensive cord compression and progressive neurological deterioration. Surgical resection and pathologic study of the mass revealed a CMF. CONCLUSIONS: A thorough Medline search has revealed that only 25 cases of spinal CMF have been reported, making this lesion an extremely rare bone tumor.

Mutations in the slow skeletal muscle fiber myosin heavy chain gene (MYH7) cause laing early-onset distal myopathy (MPD1).

We previously linked Laing-type early-onset autosomal dominant distal myopathy (MPD1) to a 22-cM region of chromosome 14. One candidate gene in the region, MYH7, which is mutated in cardiomyopathy and myosin storage myopathy, codes for the myosin heavy chain of type I skeletal muscle fibers and cardiac ventricles. We have identified five novel heterozygous mutations--Arg1500Pro, Lys1617del, Ala1663Pro, Leu1706Pro, and Lys1729del in exons 32, 34, 35, and 36 of MYH7--in six families with early-onset distal myopathy. All five mutations are predicted, by in silico analysis, to locally disrupt the ability of the myosin tail to form the coiled coil, which is its normal structure. These findings demonstrate that heterozygous mutations toward the 3' end of MYH7 cause Laing-type early-onset distal myopathy. MYH7 is the fourth distal-myopathy gene to have been identified.