Aminocarboxylic acids related to aspergillomarasmine A (AMA) and ethylenediamine-N,N'-disuccinic acid (EDDS) are strong zinc-binders and inhibitors of the metallo-beta-lactamase NDM-1

Microbial Biotechnolog

Hubble Space Telescope Grism Spectroscopy of Extreme Starbursts Across Cosmic Time: The Role of Dwarf Galaxies in the Star Formation History of the Universe

Near infrared slitless spectroscopy with the Wide Field Camera 3, onboard the Hubble Space Telescope, offers a unique opportunity to study low-mass galaxy populations at high-redshift ($z\sim$1-2). While most high$-z$ surveys are biased towards massive galaxies, we are able to select sources via their emission lines that have very-faint continua. We investigate the star formation rate (SFR)-stellar mass ($M_{\star}$) relation for about 1000 emission-line galaxies identified over a wide redshift range of $0.3 \lesssim z \lesssim 2.3$. We use the H$_{\alpha}$ emission as an accurate SFR indicator and correct the broadband photometry for the strong nebular contribution to derive accurate stellar masses down to $M_{\star} \sim 10^{7} M_{\odot}$. We focus here on a subsample of galaxies that show extremely strong emission lines (EELGs) with rest-frame equivalent widths ranging from 200 to 1500 \AA. This population consists of outliers to the normal SFR-$M_{\star}$ sequence with much higher specific SFRs ($> 10$ Gyr$^{-1}$). While on-sequence galaxies follow a continuous star formation process, EELGs are thought to be caught during an extreme burst of star formation that can double their stellar mass in less than $100$ Myr. The contribution of starbursts to the total star formation density appears to be larger than what has been reported for more massive galaxies in previous studies. In the complete mass range $8.2 <$ log($M_{\star}/M_{\odot}$) $< 10$ and a SFR lower completeness limit of about 2 $M_{\odot}$ yr$^{-1}$ (10 $M_{\odot}$ yr$^{-1}$) at $z\sim1$ ($z \sim 2$), we find that starbursts having EW$_{rest}$(H$_{\alpha}$)$>$ 300, 200, and 100 A contribute up to $\sim13$, 18, and 34 %, respectively, to the total SFR of emission-line selected sample at $z\sim1-2$. The comparison with samples of massive galaxies shows an increase in the contribution of starbursts towards lower masses.Comment: 11 pages, 6 figures. The Astrophysical Journal, in pres

Nicotinamide N-methyltransferase catalyses the N-methylation of the endogenous ß-carboline norharman: evidence for a novel detoxification pathway

Nicotinamide N-methyltransferase (NNMT) is responsible for the N-methylation of nicotinamide to 1-methylnicotinamide. Our recent studies have demonstrated that NNMT regulates cellular processes fundamental to the correct functioning and survival of the cell. It has been proposed that NNMT may possess β-carboline (BC) N-methyltransferase activity, endogenously and exogenously produced pyridine-containing compounds which, when N-methylated, are potent inhibitors of Complex I and have been proposed to have a role in the pathogenesis of Parkinson's disease. We have investigated the ability of recombinant NNMT to N-methylate norharman (NH) to 2-N-methylnorharman (MeNH). In addition, we have investigated the toxicity of the BC NH, its precursor 1,2,3,4-tetrahydronorharman (THNH) and its N-methylated metabolite MeNH, using our in vitro SH-SY5Y NNMT expression model. Recombinant NNMT demonstrated NH 2N-methyltransferase activity, with a Km of 90 ± 20 µM, a kcat of 3 × 10(-4) ± 2 × 10(-5) s(-1) and a specificity constant (kcat/Km) of 3 ± 1 s(-1) M(-1) THNH was the least toxic of all three compounds investigated, whereas NH demonstrated the greatest, with no difference observed in terms of cell viability and cell death between NNMT-expressing and non-expressing cells. In NNMT-expressing cells, MeNH increased cell viability and cellular ATP concentration in a dose-dependent manner after 72 and 120 h incubation, an effect that was not observed after 24 h incubation or in non-NNNT-expressing cells at any time point. Taken together, these results suggest that NNMT may be a detoxification pathway for BCs such as NH

Staphylococcus aureus sortase a-mediated incorporation of peptides: Effect of peptide modification on incorporation

The endogenous Staphylococcus aureus sortase A (SrtA) transpeptidase covalently anchors cell wall-anchored (CWA) proteins equipped with a specific recognition motif (LPXTG) into the peptidoglycan layer of the staphylococcal cell wall. Previous in situ experiments have shown that SrtA is also able to incorporate exogenous, fluorescently labelled, synthetic substrates equipped with the LPXTG motif (K(FITC)LPETG-amide) into the bacterial cell wall, albeit at high concentrations of 500 μM to 1 mM. In the present study, we have evaluated the effect of substrate modification on the incorporation efficiency. This revealed that (i) by elongation of LPETG-amide with a sequence of positively charged amino acids, derived from the C-terminal domain of physiological SrtA substrates, the incorporation efficiency was increased by 20-fold at 10 μM, 100 μM and 250 μM; (ii) Substituting aspartic acid (E) for methionine increased the incorporation of the resulting K(FITC)LPMTG-amide approximately three times at all concentrations tested; (iii) conjugation of the lipid II binding antibiotic vancomycin to K(FITC)LPMTG-amide resulted in the same incorporation levels as K(FITC)LPETG-amide, but much more efficient at an impressive 500-fold lower substrate concentration. These newly developed synthetic substrates can potentially find broad applications in for example the in situ imaging of bacteria; the incorporation of antibody recruiting moieties; the targeted delivery and covalent incorporation of antimicrobial compounds into the bacterial cell wall

Mechanistic insights into the C₅₅-P targeting lipopeptide antibiotics revealed by structure-activity studies and high-resolution crystal structures

The continued rise of antibiotic resistance is a global concern that threatens to undermine many aspects of modern medical practice. Key to addressing this threat is the discovery and development of new antibiotics that operate by unexploited modes of action. The so-called calcium-dependent lipopeptide antibiotics (CDAs) are an important emerging class of natural products that provides a source of new antibiotic agents rich in structural and mechanistic diversity. Notable in this regard is the subset of CDAs comprising the laspartomycins and amphomycins/friulimicins that specifically target the bacterial cell wall precursor undecaprenyl phosphate (C(55)-P). In this study we describe the design and synthesis of new C(55)-P-targeting CDAs with structural features drawn from both the laspartomycin and amphomycin/friulimicin classes. Assessment of these lipopeptides revealed previously unknown and surprisingly subtle structural features that are required for antibacterial activity. High-resolution crystal structures further indicate that the amphomycin/friulimicin-like lipopeptides adopt a unique crystal packing that governs their interaction with C(55)-P and provides an explanation for their antibacterial effect. In addition, live-cell microscopy studies provide further insights into the biological activity of the C(55)-P targeting CDAs highlighting their unique mechanism of action relative to the clinically used CDA daptomycin

Probing the Inhibitor versus Chaperone Properties of sp2-Iminosugars towards Human β-Glucocerebrosidase: A Picomolar Chaperone for Gaucher Disease

A series of sp2-iminosugar glycomimetics differing in the reducing or nonreducing character, the configurational pattern (d-gluco or l-ido), the architecture of the glycone skeleton, and the nature of the nonglycone substituent has been synthesized and assayed for their inhibition properties towards commercial glycosidases. On the basis of their affinity and selectivity towards GH1 β-glucosidases, reducing and nonreducing bicyclic derivatives having a hydroxylation profile of structural complementarity with d-glucose and incorporating an N′-octyl-isourea or -isothiourea segment were selected for further evaluation of their inhibitory/chaperoning potential against human glucocerebrosidase (GCase). The 1-deoxynojirimycin (DNJ)-related nonreducing conjugates behaved as stronger GCase inhibitors than the reducing counterparts and exhibited potent chaperoning capabilities in Gaucher fibroblasts hosting the neuronopathic G188S/G183W mutation, the isothiourea derivative being indeed one of the most efficient chaperone candidates reported up to date (70% activity enhancement at 20 pM). At their optimal concentration, the four selected compounds promoted mutant GCase activity enhancements over 3-fold; yet, the inhibitor/chaperoning balance became unfavorable at much lower concentration for nonreducing as compared to reducing derivatives.España Ministerio de Economía y Competitividad (contract numbers CTQ2015-64425-C2-1-R and SAF2016-76083-R)Junta de Andalucía contract number FQM2012-146

Probing the Inhibitor versus Chaperone Properties of sp2-Iminosugars towards Human -Glucocerebrosidase: A Picomolar Chaperone for Gaucher Disease

A series of sp2-iminosugar glycomimetics differing in the reducing or nonreducing character, the configurational pattern (d-gluco or l-ido), the architecture of the glycone skeleton, and the nature of the nonglycone substituent has been synthesized and assayed for their inhibition properties towards commercial glycosidases. On the basis of their affinity and selectivity towards GH1 β-glucosidases, reducing and nonreducing bicyclic derivatives having a hydroxylation profile of structural complementarity with d-glucose and incorporating an N′-octyl-isourea or -isothiourea segment were selected for further evaluation of their inhibitory/chaperoning potential against human glucocerebrosidase (GCase). The 1-deoxynojirimycin (DNJ)-related nonreducing conjugates behaved as stronger GCase inhibitors than the reducing counterparts and exhibited potent chaperoning capabilities in Gaucher fibroblasts hosting the neuronopathic G188S/G183W mutation, the isothiourea derivative being indeed one of the most efficient chaperone candidates reported up to date (70% activity enhancement at 20 pM). At their optimal concentration, the four selected compounds promoted mutant GCase activity enhancements over 3-fold; yet, the inhibitor/chaperoning balance became unfavorable at much lower concentration for nonreducing as compared to reducing derivatives.Ministerio de Economía y Competitividad CTQ2015-64425-C2-1-R , SAF2016-76083-RJunta de Andalucía FQM2012-1467Japan Society for the Promotion of Science JSPS KAKENHI 17K1005

Quantification of Protein Glycosylation Using Nanopores

Although nanopores can be used for singlemolecule sequencing of nucleic acids using low-cost portable devices, the characterization of proteins and their modifications has yet to be established. Here, we show that hydrophilic or glycosylated peptides translocate too quickly across FraC nanopores to be recognized. However, high ionic strengths (i.e., 3 M LiCl) and low pH (i.e., pH 3) together with using a nanopore with a phenylalanine at its constriction allows the recognition of hydrophilic peptides, and to distinguish between mono- and diglycosylated peptides. Using these conditions, we devise a nanopore method to detect, characterize, and quantify posttranslational modifications in generic proteins, which is one of the pressing challenges in proteomic analysis

Численный анализ распространения и усиления волн цунами на северо-западном шельфе Черного моря

Выполнен численный анализ распространения длинных волн в северо-западной части Черного моря. Рассмотрено 10 возможных зон сейсмической генерации цунами. Расчеты выполнены на сетке с шагом 500 м. Показано, что положение очага цунами существенно влияет на распределение высот волн вдоль побережья. Как правило, наиболее интенсивные волны формируются у ближайшего участка берега. Землетрясения в Южнобережной сейсмической зоне не могут привести к цунамиопасности в западной части моря. Только сильные землетрясения в северо-западной части способны вызывать заметные колебания уровня Черного моря. Период цунами в районе Одессы составляет около 1 ч и зависит от магнитуды землетрясения, в районе Севастополя он в 2 – 3 раза меньше. В большинстве пунктов побережья экстремальные подъемы и понижения уровня моря не превышают по абсолютной величине начального смещения поверхности моря в очаге цунами. Для отдельных участков побережья Румынии и западного побережья Крыма наблюдается некоторое усиление волн, излученных из зон генерации, расположенных в более глубоководной части исследуемого района. С ростом магнитуды землетрясения усиление волн у берега становится более значительным.Виконаний чисельний аналіз розповсюдження довгих хвиль у північно-західній частині Чорного моря. Розглянуто 10 можливих зон сейсмічної генерації цунамі. Розрахунки виконані на сітці з кроком 500 м. Показано, що положення осередку цунамі суттєво впливає на розподіл висот хвиль уздовж побережжя. Як правило, найінтенсивніші хвилі формуються близько найближчої ділянки берега. Землетруси в південнобережній сейсмічній зоні не можуть призвести до цунамонебезпеки в західній частині моря. Лише сильні землетруси в північнозахідній частині здатні викликати помітні коливання рівня Чорного моря. Період цунамі в районі Одеси складає близько 1 години і залежить від магнітуди землетрусу, в районі Севастополя він в 2 – 3 рази менший. У більшості пунктів побережжя екстремальні підйоми і пониження рівня моря не перевищують за абсолютною величиною початкового зсуву поверхні моря в осередку цунамі. Для окремих ділянок побережжя Румунії і західного побережжя Криму спостерігається деяке посилення хвиль, які випромінюють із зон генерації, розташованих в більш глибоководній частині досліджуваного району. Із зростанням магнітуди землетрусу посилення хвиль біля берега стає значнішим.Numerical analysis of long wave propagation in the Black Sea northwestern part is carried out. Ten possible zones of tsunami seismic generation are considered. The calculation are performed on the grid with a step 500 m. It is shown that location of tsunami source effects essentially the distribution of waves’ heights along the coast. As a rule, the most intensive waves are formed in the part closest to the coast. Earthquakes in the South coast seismic zone can not result in tsunami threat in the western part of the sea. Only strong earthquakes in the Black Sea northwestern part can generate noticeable sea level oscillations. Tsunami period near Odessa is about one hour and it depends on the earthquake magnitude. In the Sevastopol region it is 2 – 3 times lower. In the majority of coastal points extreme rises and falls of the sea level do not exceed the absolute value of the initial sea surface elevation in the tsunami source. Some intensification of the waves generated in deeper regions of the area under study is possible in certain parts of the Romanian coast and the Crimean western coast. The wave intensification near the coast grows with the increase of the earthquake magnitude

Synthesis and Evaluation of Polymyxins Bearing Reductively Labile Disulfide-Linked Lipids

Polymyxins are a class of lipopeptide anti-infective agents with potent and specific activity against Gram-negative bacteria. While toxicity concerns associated with polymyxin B and E (colistin) have historically limited their clinical application, today they are increasingly used as last-resort antibiotics given the rise of multidrug-resistant Gram-negative pathogens. The adverse side effects of polymyxins are well known, particularly as related to their nephrotoxicity. Here, we describe the synthesis and evaluation of a novel series of polymyxin analogues, aimed at reducing their nephrotoxic effects. Using a semisynthetic approach, we explored modifications of the exocyclic part of the polymyxin scaffold, namely, the terminal amino acid and lipophilic tail. By incorporating a reductively labile disulfide linkage in the lipid tail, we obtained novel polymyxins that exhibit potent antibacterial activity on par with polymyxin B but with reduced toxicity toward human renal proximal tubular epithelial cells