1,3-Dipolar Cycloaddition between Dehydroalanines and C,N-Cyclic Azomethine Imines: Application to Late-Stage Peptide Modification.

A non-catalytic, mild, and easy-to-handle protecting group switched 1,3-dipolar cycloaddition (1,3-DC) between bi- or mono-N-protected Dha and C,N-cyclic azomethine imines, which afford various quaternary amino acids with diverse scaffolds, is disclosed. Specifically, normal-electron-demand 1,3-DC reaction occurs between bi-N-protected Dha and C,N-cyclic azomethine imines, while inverse-electron-demand 1,3-DC reaction occurs between mono-N-protected Dha and C,N-cyclic azomethine imines. Above all, the reactions can be carried out between peptides with Dha residues at the position of interest and C,N-cyclic azomethine imines, both in homogeneous phase and on resins in SPPS. It provides a new toolkit for late-stage peptide modification, labeling, and peptide-drug conjugation. To shed light on the high regioselectivity of the reaction, DFT calculations were carried out, which were qualitatively consistent with the experimental observations.

Asymmetric N-aminoalkylation of 3-substituted indoles by N-protected N,O-acetals: an access to chiral propargyl aminals.

A direct enantioselective N1 aminoalkylation of 3-substituted indoles is efficiently catalyzed by a phosphoric acid catalyst under mild conditions to afford diverse enantioenriched propargyl aminals. The strategy could be applied to the modification of tryptophan containing oligopeptides. Additionally, structurally diverse and multifunctional transformations of the propargyl aminal products reveal the potential synthetic utility of this protocol.

Organocatalytic Enantioselective Synthesis of Tetrasubstituted α-Amino Allenoates by Dearomative γ-Addition of 2,3-Disubstituted Indoles to ß,γ-Alkynyl-α-imino Esters.

The first asymmetric synthesis of tetrasubstituted α-amino allenoates by a chiral phosphoric acid catalyzed dearomative γ-addition reaction of 2,3-disubstituted indoles to ß,γ-alkynyl-α-imino esters is reported. This method provides access to a series of highly functionalized tetrasubstituted allenes featuring quaternary stereocenters in high yields, and with excellent regio-, diastereo-, and enantioselectivities under mild conditions without by-product formation. Representative large-scale reactions and diverse transformations of the products into various scaffolds with potential biological activities render are also disclosed. The mechanism of the reaction was elucidated by control reactions and DFT calculations.

Predictive value of hyponatremia for short-term mortality in supratentorial spontaneous intracerebral hemorrhage: a single center study.

Background: Hyponatremia is a common electrolyte disturbance in patients with neurological disease; however, its predictive role for outcome in patients with supratentorial spontaneous intracerebral hemorrhage (sICH) is controversial. This study aims to explore the association between hyponatremia within 7 days after bleeding and 90-day mortality in patients with supratentorial sICH. Methods: A retrospective analysis was conducted at our institution. Patients with sICH meeting the inclusion criteria were enrolled in this study. Multivariate regression analyses were performed to determine the predictive value of hyponatremia (serum sodium <135 mmol/L) for 90-day mortality and functional outcome. Subgroup analysis was performed based on the degree and duration of hyponatremia and therapeutic strategies. The Spearman correlation test was performed to explore the relationship between hyponatremia severity and duration with variables in a multivariate regression model. Kaplan-Meier curve was depicted to reveal the relationship between hyponatremia and mortality. The receiver operating characteristic (ROC) curve was plotted to show the diagnostic effect of the minimum concentration of serum sodium (sodiummin) on 90-day mortality. Results: A total of 960 patients were enrolled, 19.6% (188) of whom were patients with hyponatremia and 26.0% (250) had 90-day mortality. The incidence of hyponatremia was roughly 2.5 times in non-survivors compared with survivors (34.8% vs. 14.2%). Multivariate regression analysis revealed that hyponatremia was the independent predictor of 90-day mortality (OR 2.763, 95%CI 1.836-4.157) and adverse outcome (OR 3.579, 95%CI 2.332-6.780). Subgroup analysis indicated an increased trend in mortality risk with both duration (more or less than 48 h) and severity of hyponatremia (mild, moderate, and severe) and confirmed the predictive value of hyponatremia for mortality in patients undergoing surgical intervention (external ventricular drainage, craniotomy, and decompressive craniectomy; all p < 0.05). The Spearman correlation test indicated no moderate or strong relationship between hyponatremia severity and duration with other variables in the multivariate model (all |rs| < 0.4). The ROC curve suggested the moderate diagnostic performance of sodiummin for mortality in both general patients and subgroups of therapeutic method patients (AUC from 0.6475 to 0.7384). Conclusion: Hyponatremia occurring in the first 7 days after bleeding is an independent predictor of 90-day morality and adverse outcome. Rigorous electrolyte scrutiny in patients treated surgically is required.

Late-stage peptide modification and macrocyclization enabled by tertiary amine catalyzed tryptophan allylation.

Late-stage modification of peptides could potentially endow peptides with significant bioactivity and physicochemical properties, and thereby provide novel opportunities for peptide pharmaceutical studies. Since tryptophan (Trp) bears a unique indole ring residue and plays various critical functional roles in peptides, the modification methods for tryptophan were preliminarily developed with considerable progress via transition-metal mediated C-H activation. Herein, we report an unprecedented tertiary amine catalyzed peptide allylation via the SN2'-SN2' pathway between the N1 position of the indole ring of Trp and Morita-Baylis-Hillman (MBH) carbonates. Using this method that proceeds under mild conditions, we demonstrated an extremely broad scope of Trp-containing peptides and MBH carbonates to prepare a series of peptide conjugates and cyclic peptides. The reaction is amenable to either solid-phase (on resin) or solution-phase conditions. In addition, the modified peptides can be further conjugated with other biomolecules at Trp, providing a new handle for bioconjugation.

Orthogonal bioconjugation targeting cysteine-containing peptides and proteins using alkyl thianthrenium salts.

Late-stage specific and selective diversifications of peptides and proteins performed at target residues under ambient conditions are recognized to be the most facile route to various and abundant conjugates. Herein, we report an orthogonal modification of cysteine residues using alkyl thianthreium salts, which proceeds with excellent chemoselectivity and compatibility under mild conditions, introducing a diverse array of functional structures. Crucially, multifaceted bioconjugation is achieved through clickable handles to incorporate structurally diverse functional molecules. This "two steps, one pot" bioconjugation method is successfully applied to label bovine serum albumin. Therefore, our technique is a versatile and powerful tool for late-stage orthogonal bioconjugation.

Cysteine-Specific Multifaceted Bioconjugation of Peptides and Proteins Using 5-Substituted 1,2,3-Triazines.

Peptide and protein postmodification have gained significant attention due to their extensive impact on biomolecule engineering and drug discovery, of which cysteine-specific modification strategies are prominent due to their inherent nucleophilicity and low abundance. Herein, the study introduces a novel approach utilizing multifunctional 5-substituted 1,2,3-triazine derivatives to achieve multifaceted bioconjugation targeting cysteine-containing peptides and proteins. On the one hand, this represents an inaugural instance of employing 1,2,3-triazine in biomolecular-specific modification within a physiological solution. On the other hand, as a powerful combination of precision modification and biorthogonality, this strategy allows for the one-pot dual-orthogonal functionalization of biomolecules utilizing the aldehyde group generated simultaneously. 1,2,3-Triazine derivatives with diverse functional groups allow conjugation to peptides or proteins, while bi-triazines enable peptide cyclization and dimerization. The examination of the stability of bi-triazines revealed their potential for reversible peptide modification. This work establishes a comprehensive platform for identifying cysteine-selective modifications, providing new avenues for peptide-based drug development, protein bioconjugation, and chemical biology research.

Current understanding of macrophages in intracranial aneurysm: relevant etiological manifestations, signaling modulation and therapeutic strategies.

Macrophages activation and inflammatory response play crucial roles in intracranial aneurysm (IA) formation and progression. The outcome of ruptured IA is considerably poor, and the mechanisms that trigger IA progression and rupture remain to be clarified, thereby developing effective therapy to prevent subarachnoid hemorrhage (SAH) become difficult. Recently, climbing evidences have been expanding our understanding of the macrophages relevant IA pathogenesis, such as immune cells population, inflammatory activation, intra-/inter-cellular signaling transductions and drug administration responses. Crosstalk between macrophages disorder, inflammation and cellular signaling transduction aggravates the devastating consequences of IA. Illustrating the pros and cons mechanisms of macrophages in IA progression are expected to achieve more efficient treatment interventions. In this review, we summarized the current advanced knowledge of macrophages activation, infiltration, polarization and inflammatory responses in IA occurrence and development, as well as the most relevant NF-κB, signal transducer and activator of transcription 1 (STAT1) and Toll-Like Receptor 4 (TLR4) regulatory signaling modulation. The understanding of macrophages regulatory mechanisms is important for IA patients' clinical outcomes. Gaining insight into the macrophages regulation potentially contributes to more precise IA interventions and will also greatly facilitate the development of novel medical therapy.

Visible-Light Mediated Deoxygenation of Carboxylic Acid for Late-Stage Peptide Modification Targeting Dehydroalanine.

A visible-light mediated deoxygenative radical addition of carboxylic acids to dehydroalanines has been disclosed. The method can be used in ß-acyl alanine derivative synthesis, including those chiral and deuterated variants, and late-stage peptide modification with various functional groups, both in the homogeneous phase and on the resin in SPPS. It provides a new tool kit for rapid construction of bioactive peptide analogues, which has been demonstrated by modification of the antimicrobial peptide Feleucin-K3.

NaClO-Mediated Cross Installation of Indoles and Azoles Benefits Anticancer Hit Discovery.

Innovations in synthetic chemistry have a profound impact on the drug discovery process, and will always be a necessary driver of drug development. As a result, it is of significance to develop novel simple and effective synthetic installation of medicinal modules to promote drug discovery. Herein, we have developed a NaClO-mediated cross installation of indoles and azoles, both of which are frequently encountered in drugs and natural products. This effective toolbox provides a convenient synthetic route to access a library of N-linked 2-(azol-1-yl) indole derivatives, and can be used for late-stage modification of drugs, natural products and peptides. Moreover, biological screening of the library has revealed that several adducts showed promising anticancer activities against A549 and NCI-H1975 cells, which give us a hit for anticancer drug discovery.

Single-cell analysis of peripheral blood from high-altitude pulmonary hypertension patients identifies a distinct monocyte phenotype.

Immune and inflammatory responses have an important function in the pathophysiology of pulmonary hypertension (PH). However, little is known about the immune landscape in peripheral circulation in patients with high-altitude pulmonary hypertension (HAPH). We apply single-cell transcriptomics to characterize the monocytes that are significantly enriched in the peripheral blood mononuclear cells (PBMC) of HAPH patients. We discover an increase in C1 (non-classical) and C2 (intermediate) monocytes in PBMCs and a decrease in hypoxia-inducible transcription factor-1α (HIF-1α) in all monocyte subsets associated with HAPH. In addition, we demonstrate that similar immune adaptations may exist in HAPH and PH. Overall, we characterize an immune cell atlas of the peripheral blood in HAPH patients. Our data provide evidence that specific monocyte subsets and HIF-1α downregulation might be implicated in the pathogenesis of HAPH.

On rigid origami III: local rigidity analysis.

In this article, rigid origami is examined from the perspective of rigidity theory. First- and second-order rigidity are defined from local differential analysis of the consistency constraint; while the static rigidity and prestress stability are defined after finding the form of internal force and load. We will show the hierarchical relation among these local rigidities with examples representing different levels. The development of theory here follows the same path as the conventional rigidity theory for bar-joint frameworks, but starts with different high-order rotational constraints. We also bring new interpretation to the internal force and geometric error of constraints associated with energy. Examining the different aspects of the rigidity of origami might give a novel perspective for the development of new folding patterns, or for the design of origami structures where some rigidity is required.

Copper(I)-Catalyzed Late-Stage Introduction of Oxime Ethers into Peptides at the Carboxylic Acid Site.

We have developed a method of introducing biological oxime ether fragments into peptides by CuI-catalyzed late-stage modification and functionalization of peptides, utilizing their acid moiety and varied 2H-azirines. As a result of its mild conditions, high atom economy, moderate yield, and excellent functional-group tolerance, the method can provide access to late-stage peptide modification and functionalization at their acid sites both in the homogeneous phase and on resins in SPPS, providing a new tool kit for peptide functionalization, diversification, and fluorescent labeling.

NDTP Mediated Direct Rapid Amide and Peptide Synthesis without Epimerization.

Herein, we explored an unprecedented mild, nonirritating, conveniently available, and recyclable coupling reagent NDTP, which could activate the carboxylic acids via acyl thiocyanide and enable the rapid amide and peptide synthesis at very mild conditions. In addition, the methodology was compatible with Fmoc-SPPS, which may provide an alternative to peptide manufacturing.

Access to α,γ-Diamino Diacid Derivatives via Organocatalytic Asymmetric 1,4-Addition of Azlactones and Dehydroalanines.

A convenient and functional-group-tolerant organocatalytic asymmetric 1,4-addition of azlactones and dehydroalanine is disclosed. The reaction is used for the first synthesis of chiral α,γ-diamino diacid derivatives with nonadjacent stereogenic centers in moderate to high yields, with excellent diastereo- and enantioselectivities, under the catalysis of a chiral thiourea catalyst. In addition, the reaction could be conducted in gram-scale, and the products of the reaction could be readily converted to various α,γ-diamino diacid derivatives, α,γ-diamino dialcohols, and modified peptides with nonproteinogenic amino acid residues.

[Infection of Helicobacter pylori strains with multiple vacA m-region subtypes and/or iceA mixed genotype among the Bai, Naxi, and Han populations in Yunnan province].

OBJECTIVE: To study the epidemiology and pathogenicity of mixed infection of Helicobacter pylori (H. pylori) strains with multiple vacA m-region subtypes and/or iceA mixed genotype among the Bai, Naxi, and Han populations in Yunnan province. METHODS: Gastric mucous membrane were obtained by gastroscopy from 444 patients with digestive ulcer or chronic gastritis, 165 of Han nationality, 117 of Bai nationality, and 162 of Naxi nationality. H. pylori was isolated from 109 patients and cultured. RT-PCR was used to detect the Helicobacter pylori strains vacA gene subtype and iceA genes. RESULTS: The overall rates of vacA gene s1 type and vacA gene s2 type were 98.2% (107/109) and 1.8% (2/109) respectively. The overall rates of vacA gene m2 type, vacA gene m1a type, vacA gene m1b type, and mixed types were 45.9% (50/109), 0.9% (1.109), 23.9% (26/109), and 11% (12/109) respectively; and m region was not identified in 18.3% (20/109) of the strains. The overall rates of iceA1 and iceA2 genes were 67.0% (73/109) and 41.3% (45/109) respectively. In the specimens from the patients of Bai nationality, as regards the s region, only s1 type was identified in all specimens; the rates of vacA gene m2 type, vacA gene m1a type, vacA gene m1b type, and mixed type were 45.2% (14/31), 3.2% (1/31), 12.9% (4/31), and 16.2% (5/31) respectively, and m region was not identified in 22.6% of the strains; and the rates of iceA1 and iceA2 were 87.1% (27/31) and 61.3% (19/31) respectively. In the specimens from the patients of Naxi nationally, The rates of vacA gene s1 type and s2 type were 95.6% (43/45) and 4.4% (2/45), the rates of vacA gene m2 type, vacA gene m1b type, and mixed m-type were 33.3% (15/45), 37.8% (17/45), and 6.7% (3/45) respectively, no m1a type was found, and m region was not identified in 22.2% of the strains; the rates of iceA1 and iceA2 were 48.9% (22/45) and 31.1% (14/45) respectively. In the specimens of the patients of Han nationality, as regards the s region, all specimens were s1 type; the rates of vacA gene m2 type, vacA gene m1b type, and mixed type were 63.6% (21/33), 15.2% (5/33), and 12.1% (4/33) respectively, and m region was not identified in 9.1% (3/33) of the strains; the rates of iceA1 and iceA2 were 72.7% (24/33) and 36.4% (12/33) respectively. 34 of the 109 patients (31.2%) suffered from mixed infection of vacA gene m mixed subtype and/or iceA mixed genotype, 64.7% of which (22/34) suffering iceA1 + iceA2 infection, 23.5% of which (8/34) being iceA + vacA mixed infection, and 11.8% of which (4/34) being vacA subtype mixed infection, with the former incidence of the former group significantly higher than those of the 2 latter groups (both P < 0.001). The mixed infection rate of the Bai nationality (67.7%, 21/31) was significantly higher than those of the Naxi nationality (20.0%, 9/45, P < 0.001) and Han nationality (12.1%, 4/33, P > 0.001) without a significant difference between the Naxi and Han nationalities (P > 0.05). The mixed infection rate of iceA1 + iceA2 of the Bai nationality (61.3%, 19/31) was significantly higher than those of the Naxi nationality (17.8%, 8/45, P < 0.001) and Han nationality (9.1%, 3/33, P < 0.001) without a significant difference between the Naxi and Han nationalities (P > 0.05). The mixed rate of the patients with digestive ulcer was 30.2% (16/53), not significantly different from that of the patients with chronic gastritis (32.1%, 18/56, P > 0.05). 16 of the 34 cases of mixed infection were patients of digestive ulcer; and 8 of the 34 cases were patients with chronic gastritis. 8 of the 34 cases (23.5%) of mixed infection were cases of infection of vacA gene m mixed subtypes and iceA mixed genotype; among which 7 were patients with digestive ulcer and 1 case was patient of chronic gastritis. Thus the rate of vacA gene m mixed subtypes and iceA mixed genotype was 43.8% (7/16) in the patients with digestive ulcer, significantly higher than in the patient with chronic gastritis (5.6%, 1/18, P = 0.014). 22 of the 34 cases of mixed infection (64.7%) suffered from infection of iceA mixed gene subtypes. Among these 22 cases 15 were patients with chronic gastritis and 7 were patients with digestive ulcer. Thus the rate of iceA mixed subtypes infection was 43.8% (7/16) in the patients with digestive ulcer, significantly lower than in the patient with chronic gastritis (83.3%, 15/18, P = 0.016). In the 21 mixed infection patients of Bai nationality, the rate of vacA gene mixed sybtype + iceA gene mixed subtypes infection was 100% in the patients with digestive ulcer (100%, 5/5), significantly higher than that of single vacA gene subtype + iceA gene mixed subtypes (37.5%, 6/16, P = 0.0258). The mixed infection rate of H. pylori strains was 32.1%. CONCLUSION: The mixed infection rate of the Bai nationality is higher than those of the Han and Naxi nationalities. Detection of mixed infection by vacA genotypes is more sensitive in Han populations. Detection of multiple infections by iceA gene mixed subtypes is more sensitive in Bai and Naxi populations.

A cluster of primary pneumonic plague transmitted in a truck cab in a new enzootic focus in China.

We investigated a cluster of five cases of severe pneumonia from one village in Yunnan Province, China. We searched for severe pneumonia in the village and hospitals. We interviewed patients and family members about exposures. We tested acute and convalescent sera for antigen and antibody of severe acute respiratory syndrome, avian influenza, and plague. The only common exposure of the five patients was riding together in the enclosed cab of a truck for 1.5 hours while taking the first patient to the hospital. Seroconversion to plague F1 antigen confirmed plague in three survivors. Unfamiliarity of clinicians with plague and lack of sputum examination, blood culture, or postmortem examination delayed the diagnosis. No plague cases occurred among family and village contacts and health care workers. High infectivity in this cluster was limited to a crowded, poorly ventilated truck.