Clinical features and subdural lesions in childhood onset Haemophilus influenzae meningitis

It becomes difficult to treat the onset of childhood Haemophilus influenza type b( Hib) meningitis due tothe appearance of b-lactamase negative ampicilin resistance( BLNAR). We investigated the clinical featuresof Hib meningitis with subdural lesions. From January 2000 to December 2006, we experienced 8 patientswith Hib meningitis. All patients were not inoculated with Hib vaccine. Five of them were onset under 1year old. Rapid latex diagnoses were made in 5 patients, among whom 4 patients showed Hib positive. Thegenotypes were determined in 4 patients with BLNAR. Combined with cefotaxime sodium (CTX) and ampicillin(ABPC) were used in 3 patients for the initial antibiotics, panipenem/betamipron( PAPM/BP) in 2,meropenem hydrate( MEPM) in 1, MEPM + ceftriaxone sodium( CTRX) in 1, and concomitant use of dexamethasonein 7 paitents. MRI showed subdural hygroma in 4 patients and subdural abscess in 4 patients.Subdural lesions appeared on Day 1 to Day 18 from the onset. Three patients with subdural abscess haveperformed subdural taps, and 2of them with difficulties after subdural taps were necessary to do oral administrationof chloramphenicol( CP).In our report, all of the patients developed subdural lesions. The development of subdural lesions cannotbe avoided with only the conventional antibiotics and dexamethasone therapy. Our cases suggested earliersubdural taps with oral administration of CP might be to improve both the general condition and control thesubdural lesions with Hib meningitis. Moreover, we should pay attention to the preventive vaccination ofHib

乳幼児期の体重増加と学童期のインスリン抵抗性との関係に関する系統的・探索的解析

第3回日本DOHaD研究会学術集会　抄録集　【ポスター発表

Antibiotic Discovery with Synthetic Fermentation: Library Assembly, Phenotypic Screening, and Mechanism of Action of Beta-Peptides Targeting Penicillin-Binding Proteins

In analogy to biosynthetic pathways leading to bioactive natural products, synthetic fermentation generates mixtures of molecules from simple building blocks under aqueous, biocompatible conditions, allowing for the resulting cultures to be directly screened for biological activity. In this work, a novel beta-peptide antibiotic was successfully identified using the synthetic fermentation platform. Phenotypic screening was carried out in an initially random fashion, allowing for simple identification of active cultures. Subsequent deconvolution, focused screening and structure-activity relationship studies led to the identification of a potent antimicrobial peptide, showing strong selectivity for our model system Bacillus subtilis over human Hek293 cells. To determine the antibacterial mechanism of action, a peptide probe bearing a photoaffinity tag was readily synthesized through the use of appropriate synthetic fermentation building blocks and utilized for target identification using a quantitative mass spectrometry-based strategy. The chemoproteomic approach led to the identification of a number of bacterial membrane proteins as prospective targets. These findings were validated through binding affinity studies with penicillin-binding protein 4 using microscale thermophoresis, with the bioactive peptide showing a dissociation constant (Kd) in the nanomolar range. Through these efforts, we provide a proof of concept for the synthetic fermentation approach presented here as a new strategy for the phenotypic discovery of novel bioactive compounds

Chemical Synthesis of Interleukin‐2 and Disulfide Stabilizing Analogues

Chemical protein synthesis allows the construction of well-defined structural variations and facilitates the development of deeper understanding of protein structure-function relationships and new protein engineering strategies. Herein, we report the chemical synthesis of interleukin-2 (IL-2) variants on a multimilligram scale and the formation of non-natural disulfide mimetics that improve stability against reduction. The synthesis was accomplished by convergent KAHA ligations; the acidic conditions of KAHA ligation proved to be valuable for the solubilization of the hydrophobic segments of IL-2. The bioactivity of the synthetic IL-2 and its analogues were shown to be equipotent to recombinant IL-2 and exhibit improved stability against reducing agents

Chemical Synthesis of Interleukin-2 and Disulfide Stabilizing Analogues

ISSN:1433-7851ISSN:1521-3773ISSN:0570-083

Antibiotic Discovery with Synthetic Fermentation: Library Assembly,Phenotypic Screening, and Mechanism of Action of β‑PeptidesTargeting Penicillin-Binding Proteins

In analogy to biosynthetic pathways leading to bioactive natural products, synthetic fermentation generates mixtures of molecules from simple building blocks under aqueous, biocompatible conditions, allowing the resulting cultures to be directly screened for biological activity. In this work, a novel β-peptide antibiotic was successfully identified using the synthetic fermentation platform. Phenotypic screening was carried out in an initially random fashion, allowing simple identification of active cultures. Subsequent deconvolution, focused screening, and structure–activity relationship studies led to the identification of a potent antimicrobial peptide, showing strong selectivity for our model system Bacillus subtilis over human HEK293 cells. To determine the antibacterial mechanism of action, a peptide probe bearing a photoaffinity tag was readily synthesized through the use of appropriate synthetic fermentation building blocks and utilized for target identification using a quantitative mass spectrometry-based strategy. The chemoproteomic approach led to the identification of a number of bacterial membrane proteins as prospective targets. These findings were validated through binding affinity studies with penicillin-binding protein 4 using microscale thermophoresis, with the bioactive peptide showing a dissociation constant (Kd) in the nanomolar range. Through these efforts, we provide a proof of concept for the synthetic fermentation approach presented here as a new strategy for the phenotypic discovery of novel bioactive compounds.ISSN:1554-8929ISSN:1554-893

Facile folding of insulin variants bearing a prosthetic C-peptide prepared by -ketoacid-hydroxylamine (KAHA) ligation

The chemical synthesis of insulin is an enduring challenge due to the hydrophobic peptide chains and construction of the correct intermolecular disulfide pattern. We report a new approach to the chemical synthesis of insulin using a short, traceless, prosthetic C-peptide that facilitates the formation of the correct disulfide pattern during folding and its removal by basic treatment. The linear precursor is assembled by an ester forming α-ketoacid-hydroxylamine (KAHA) ligation that provides access to the linear insulin precursors in good yield from two readily prepared segments. This convergent and flexible route provides access to various human, mouse, and guinea pig insulins containing a single homoserine mutation that shows no detrimental effect on the biological activities.ISSN:2041-6520ISSN:2041-653

De Novo Macrocyclic Peptide Inhibitors of Hepatitis B Virus Cellular Entry

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