Phenotypic CD8 T cell profiling in chronic hepatitis B to predict HBV-specific CD8 T cell susceptibility to functional restoration in vitro.

OBJECTIVE: Exhausted hepatitis B virus (HBV)-specific CD8 T cells in chronic HBV infection are broadly heterogeneous. Characterisation of their functional impairment may allow to distinguish patients with different capacity to control infection and reconstitute antiviral function. DESIGN: HBV dextramer+CD8 T cells were analysed ex vivo for coexpression of checkpoint/differentiation markers, transcription factors and cytokines in 35 patients with HLA-A2+chronic hepatitis B (CHB) and in 29 control HBsAg negative CHB patients who seroconverted after NUC treatment or spontaneously. Cytokine production was also evaluated in HBV peptide-stimulated T cell cultures, in the presence or absence of antioxidant, polyphenolic, PD-1/PD-L1 inhibitor and TLR-8 agonist compounds and the effect on HBV-specific responses was further validated on additional 24 HLA-A2 negative CHB patients. RESULTS: Severely exhausted HBV-specific CD8 T cell subsets with high expression of inhibitory receptors, such as PD-1, TOX and CD39, were detected only in a subgroup of chronic viraemic patients. Conversely, a large predominance of functionally more efficient HBV-specific CD8 T cell subsets with lower expression of coinhibitory molecules and better response to in vitro immune modulation, typically detected after resolution of infection, was also observed in a proportion of chronic viraemic HBV patients. Importantly, the same subset of patients who responded more efficiently to in vitro immune modulation identified by HBV-specific CD8 T cell analysis were also identified by staining total CD8 T cells with PD-1, TOX, CD127 and Bcl-2. CONCLUSIONS: The possibility to distinguish patient cohorts with different capacity to respond to immune modulatory compounds in vitro by a simple analysis of the phenotypic CD8 T cell exhaustion profile deserves evaluation of its clinical applicability.

Characterization of a Novel Capsid Assembly Modulator for the Treatment of Chronic Hepatitis B Virus Infection.

The standard of care for the treatment of chronic hepatitis B (CHB) is typically lifelong treatment with nucleos(t)ide analogs (NAs), which suppress viral replication and provide long-term clinical benefits. However, infectious virus can still be detected in patients who are virally suppressed on NA therapy, which may contribute to the failure of these agents to cure most CHB patients. Accordingly, new antiviral treatment options are being developed to enhance the suppression of hepatitis B virus (HBV) replication in combination with NAs ("antiviral intensification"). Here, we describe GS-SBA-1, a capsid assembly modulator (CAM) belonging to class CAM-E, that demonstrates potent inhibition of extracellular HBV DNA in vitro (EC50 [50% effective concentration] = 19 nM) in HBV-infected primary human hepatocytes (PHHs) as well as in vivo in an HBV-infected immunodeficient mouse model. GS-SBA-1 has comparable activities across HBV genotypes and nucleos(t)ide-resistant mutants in HBV-infected PHHs. In addition, GS-SBA-1 demonstrated in vitro additivity in combination with tenofovir alafenamide (TAF). The administration of GS-SBA-1 to PHHs at the time of infection prevents covalently closed circular DNA (cccDNA) formation and, hence, decreases HBV RNA and antigen levels (EC50 = 80 to 200 nM). Furthermore, GS-SBA-1 prevents the production of extracellular HBV RNA-containing viral particles in vitro. Collectively, these data demonstrate that GS-SBA-1 is a potent CAM that has the potential to enhance viral suppression in combination with an NA.

Catalytic, Diastereoselective 1,2-Difluorination of Alkenes.

We describe a direct, catalytic approach to the 1,2-difluorination of alkenes. The method utilizes a nucleophilic fluoride source and an oxidant in conjunction with an aryl iodide catalyst and is applicable to alkenes with all types of substitution patterns. In general, the vicinal difluoride products are produced with high diastereoselectivities. The observed sense of stereoinduction implicates anchimeric assistance pathways in reactions of alkenes bearing neighboring Lewis basic functionality.

Concise and enantioselective total synthesis of (-)-mehranine, (-)-methylenebismehranine, and related Aspidosperma alkaloids.

We report an efficient and highly stereoselective strategy for the synthesis of Aspidosperma alkaloids based on the transannular cyclization of a chiral lactam precursor. Three new stereocenters are formed in this key step with excellent diastereoselectivity due to the conformational bias of the cyclization precursor, leading to a versatile pentacyclic intermediate. A subsequent stereoselective epoxidation followed by a mild formamide reduction enabled the first total synthesis of the Aspidosperma alkaloids (-)-mehranine and (+)-(6S,7S)-dihydroxy-N-methylaspidospermidine. A late-stage dimerization of (-)-mehranine mediated by scandium trifluoromethanesulfonate completed the first total synthesis of (-)-methylenebismehranine.

Direct dehydrative N-pyridinylation of amides.

Electrophilic activation of secondary amides with trifluoromethanesulfonic anhydride in the presence of 2-fluoropyridine followed by introduction of a pyridine N-oxide derivative and warming affords the corresponding N-pyridinyl tertiary amide derivatives. A mechanism supported by in situ monitoring and deuterium labeling experiments is discussed.

Biomechanical analysis of stresses to the fifth metatarsal bone during sports maneuvers: implications for fifth metatarsal fractures.

Fifth metatarsal stress fractures are an increasing problem in elite and recreational athletic populations. One possible mechanism of injury is the many bending moments applied to the fifth metatarsal during dynamic sports maneuvers involving rapid changes in direction and speed. A potentially important bending moment is loading of the base versus the head of the fifth metatarsal, which tends to cause a bending moment along the bone. To determine which maneuver applies the greatest pressure differential between the base and head of the fifth metatarsal, 10 college-aged male athletes performed running straight, jump take-off, jump landing, cutting right, cutting left, and accelerating while plantar pressures were recorded using a Pedar insole system (Novel Electronics, Inc., St. Paul, MN). Peak pressure at the fifth metatarsal base was subtracted from the peak pressure at the fifth metatarsal head to obtain the fifth metatarsal pressure differential-a corollary to the bending moment. The greatest fifth metatarsal pressure differential was observed during acceleration maneuvers (20 + or - 13.1 N/cm(2); P < 0.0001) followed by running straight (11.6 + or - 8 N/cm(2); P < 0.0008). The other maneuvers had low pressure differentials: jump take-off (4.2 + or - 10.6 N/cm(2)), jump landing (3.7 + or - 9.2 N/cm(2)), cutting left (2.3 + or - 4.2 N/cm(2)), and cutting right (-2.1 + or - 10 N/cm(2)). It appears that acceleration maneuvers may apply the largest bending moments to the fifth metatarsal and could lead to stress fractures. Because fifth metatarsal stress fractures are associated with rapid increases in training volume, reducing the number of acceleration events may be effective in altering the balance between bone resorption and bone formation and reducing stress fracture risk. Careful planning of training programs allowing for adequate rest between intense bouts of exercise involving many acceleration maneuvers may be the best preventative measure.

(S)-(+)-1-(2-Bromo-phen-yl)ethanol.

The title compound, C(8)H(9)BrO, crystallizes with two mol-ecules in the asymmetric unit. The structure displays O-H⋯O hydrogen bonding, generating zigzag chains evolving around a screw axis along [100].

Catalytic, asymmetric difluorination of alkenes to generate difluoromethylated stereocenters.

Difluoromethyl groups possess specific steric and electronic properties that invite their use as chemically inert surrogates of alcohols, thiols, and other polar functional groups important in a wide assortment of molecular recognition processes. We report here a method for the catalytic, asymmetric, migratory geminal difluorination of ß-substituted styrenes to access a variety of products bearing difluoromethylated tertiary or quaternary stereocenters. The reaction uses commercially available reagents (m-chloroperbenzoic acid and hydrogen fluoride pyridine) and a simple chiral aryl iodide catalyst and is carried out readily on a gram scale. Substituent effects and temperature-dependent variations in enantioselectivity suggest that cation-π interactions play an important role in stereodifferentiation by the catalyst.

Synthesis of spirocyclic indolines by interruption of the Bischler-Napieralski reaction.

The development of a versatile method for the synthesis of spirocyclic pyrrolidinoindolines is discussed. Treatment of N-acyltryptamines with trifluoromethanesulfonic anhydride-2-chloropyridine reagent combination affords highly persistent spiroindoleninium ions that are subject to intra- and intermolecular addition at C2 by nucleophiles.

Robinson's landmark synthesis of tropinone.

The 1917 total synthesis of tropinone by Sir Robert Robinson represents a landmark achievement in organic synthesis. Decades ahead of its time in terms of its retrosynthetic logic and biomimetic approach, the elegant combination of these two elements in this synthesis continues to serve as an inspiration for the development of new and efficient strategies for complex molecule synthesis.

Regional foot pressure during running, cutting, jumping, and landing.

BACKGROUND: Evaluating shoes during sport-related movements may provide a better assessment of plantar loads associated with repetitive injury and provide more specific data for comparing shoe cushioning characteristics. HYPOTHESIS: Accelerating, cutting, and jumping pressures will be higher than in straight running, differentiating regional shoe cushioning performance in sport-specific movements. STUDY DESIGN: Controlled laboratory study. MATERIALS AND METHODS: Peak pressures on seven anatomic regions of the foot were assessed in 10 male college athletes during running straight ahead, accelerating, cutting left, cutting right, jump take-off, and jump landing wearing Speed TD and Air Pro Turf Low shoes (Nike, Beaverton, Ore). Pedar insoles (Novel, Munich, Germany) were sampled at 99 Hz during the 6 movements. RESULTS: Cutting and jumping movements demonstrated more than double the pressure at the heel compared with running straight, regardless of shoe type. The Air Pro Turf showed overall lower pressure for all movement types (P<.0377). Cutting to the left, the Air Pro Turf shoe had lower heel pressures (36.6 +/- 12.5 N/cm(2)) than the Speed TD (50.3 +/- 11.2 N/cm(2)) (P<.0001), and the Air Pro Turf had lower great toe pressures than the Speed TD (44.8 +/- 8.1 N/cm(2) vs 54.4 +/- 8.4 N/cm(2); P= .0002). The Air Pro Turf also had significantly lower pressures than the Speed TD at the central forefoot during acceleration (38.2 +/- 8.3 N/cm(2) vs 50.8 +/- 7.4 N/cm(2); P<.0001). CONCLUSION: Sport-related movements load the plantar surface of the foot more than running straight. Shoe cushioning characteristics were more robustly assessed during sport-related movements (4 significant results detected) compared with running straight (1 significant result detected). CLINICAL RELEVANCE: There is an interaction between shoe cushioning characteristics and sport-related movements that may influence plantar pressure and repetitive stress injuries.