Enzymatical and microbial degradation of cyclic dipeptides (diketopiperazines)

Diketopiperazines (DKPs) are cyclic dipeptides, representing an abundant class of biologically active natural compounds. Despite their widespread occurrence in nature, little is known about their degradation. In this study, the enzymatical and microbial cleavage of DKPs was investigated. Peptidase catalyzed hydrolysis of certain DKPs was formerly reported, but could not be confirmed in this study. While testing additional peptidases and DKPs no degradation was detected, indicating peptidase stability of the peptide bond in cyclic dipeptides. Besides confirmation of the reported degradation of cyclo(l-Asp-l-Phe) by Paenibacillus chibensis (DSM 329) and Streptomyces flavovirens (DSM 40062), cleavage of cyclo(l-Asp-l-Asp) by DSM 329 was detected. Other DKPs were not hydrolyzed by both strains, demonstrating high substrate specificity. The degradation of cyclo(l-Asp-l-Phe) by DSM 40062 was shown to be inducible. Three strains, which are able to hydrolyze hydantoins and dihydropyrimidines, were identified for the degradation of DKPs: Leifsonia sp. K3 (DSM 27212) and Bacillus sp. A16 (DSM 25052) cleaved cyclo(dl-Ala-dl-Ala) and cyclo(l-Gly-l-Phe), and Rhizobium sp. NA04-01 (DSM 24917) degraded cyclo(l-Asp-l-Phe), cyclo(l-Gly-l-Phe) and cyclo(l-Asp-l-Asp). The first enantioselective cleavage of cyclo(dl-Ala-dl-Ala) was detected with the newly isolated strains Paenibacillus sp. 32A (DSM 27214) and Microbacterium sp. 40A (DSM 27211). Cyclo(l-Ala-d-Ala) and cyclo(l-Ala-l-Ala) were completely degraded, whereas the enantiomer cyclo(d-Ala-d-Ala) was not attacked. Altogether, five bacterial strains were newly identified for the cleavage of DKPs. These bacteria may be of value for industrial purposes, such as degradation of undesirable DKPs in food and drugs and production of (enantiopure) dipeptides and amino acids

Novel amidases of two Aminobacter sp. strains: Biotransformation experiments and elucidation of gene sequences

The amidase activities of two Aminobacter sp. strains (DSM24754 and DSM24755) towards the aryl-substituted substrates phenylhydantoin, indolylmethyl hydantoin, D,L-6-phenyl-5,6-dihydrouracil (PheDU) and para-chloro-D,L-6-phenyl-5,6-dihydrouracil were compared. Both strains showed hydantoinase and dihydropyrimidinase activity by hydrolyzing all substrates to the corresponding N-carbamoyl-α- or N-carbamoyl-β-amino acids. However, carbamoylase activity and thus a further degradation of these products to α- and β-amino acids was not detected. Additionally, the genes coding for a dihydropyrimidinase and a carbamoylase of Aminobacter sp. DSM24754 were elucidated. For Aminobacter sp. DSM24755 a dihydropyrimidinase gene flanked by two genes coding for putative ABC transporter proteins was detected. The deduced amino acid sequences of both dihydropyrimidinases are highly similar to the well-studied dihydropyrimidinase of Sinorhizobium meliloti CECT4114. The latter enzyme is reported to accept substituted hydantoins and dihydropyrimidines as substrates. The deduced amino acid sequence of the carbamoylase gene shows a high similarity to the very thermostable enzyme of Pseudomonas sp. KNK003A

‘They’re more like ordinary stroppy British women’: Attitudes and expectations of maternity care professionals to UK-born ethnic minority women

Objective To explore the attitudes and expectations of maternity care professionals to UK-born ethnic minority mothers. Methods Qualitative in-depth interviews with 30 professionals from eight NHS maternity units in England that provide services for large proportions of women of black Caribbean, black African, Indian, Pakistani and Irish descent. Results All the professionals reported providing care to both UK-born and migrant mothers from ethnic minorities. Most of them felt that they could differentiate between UK-born and migrant mothers based mainly on language fluency and accent. ‘Westernized dress’ and ‘freedom’ were also cited as indicators. Overall, professionals found it easier to provide services to UK-born mothers and felt that their needs were more like those of white English mothers than those of migrant mothers. UK-born mothers were generally thought to be assertive and expressive, and in control of care-related decision-making whereas some South Asian Muslim women were thought to be constrained by family influences. Preconceived ideas about ethnic minority mothers' tolerance of pain in labour, use of pharmacological pain relief measures and mode of delivery were recurring themes. Women's education and social class were felt to be major influences on the uptake of maternity care, regardless of ethnicity. Conclusions Professionals appeared to equate the needs of UK-born ethnic minority women with those of white English women. Overall, this has positive implications for care provision. Despite this, specific behavioural expectations and unconscious stereotypical views were evident and have the potential to affect clinical practice

Upk3b is dispensable for development and integrity of urothelium and mesothelium

The mesothelium, the lining of the coelomic cavities, and the urothelium, the inner lining of the urinary drainage system, are highly specialized epithelia that protect the underlying tissues from mechanical stress and seal them from the overlying fluid space. The development of these epithelia from simple precursors and the molecular characteristics of the mature tissues are poorly analyzed. Here, we show that uroplakin 3B (Upk3b), which encodes an integral membrane protein of the tetraspanin superfamily, is specifically expressed both in development as well as under homeostatic conditions in adult mice in the mesothelia of the body cavities, i.e., the epicardium and pericardium, the pleura and the peritoneum, and in the urothelium of the urinary tract. To analyze Upk3b function, we generated a creERT2 knock-in allele by homologous recombination in embryonic stem cells. We show that Upk3bcreERT2 represents a null allele despite the lack of creERT2 expression from the mutated locus. Morphological, histological and molecular analyses of Upk3b-deficient mice did not detect changes in differentiation or integrity of the urothelium and the mesothelia that cover internal organs. Upk3b is coexpressed with the closely related Upk3a gene in the urothelium but not in the mesothelium, leaving the possibility of a functional redundancy between the two genes in the urothelium only

Mass and density of the transiting hot and rocky super-Earth LHS 1478 b (TOI-1640 b)

One of the main objectives of the Transiting Exoplanet Survey Satellite ({TESS}) mission is the discovery of small rocky planets around relatively bright nearby stars. Here, we report the discovery and characterization of the transiting super-Earth planet orbiting LHS~1478 (TOI-1640). The star is an inactive red dwarf ($J \sim 9.6$\,mag and spectral type m3\,V) with mass and radius estimates of $0.20\pm0.01$\,$M_{\odot}$ and $0.25\pm0.01$\,$R_{\odot}$, respectively, and an effective temperature of $3381\pm54$\,K.It was observed by \tess in four sectors. These data revealed a transit-like feature with a period of 1.949 days. We combined the TESS data with three ground-based transit measurements, 57 radial velocity (RV) measurements from CARMENES, and 13 RV measurements from IRD, determining that the signal is produced by a planet with a mass of $2.33^{+0.20}_{-0.20}$\,$M_{\oplus}$ and a radius of $1.24^{+0.05}_{-0.05}$\,$R_{\oplus}$. The resulting bulk density of this planet is 6.67\,g\,cm$^{-3}$, which is consistent with a rocky planet with an Fe- and MgSiO$_3$-dominated composition. Although the planet would be too hot to sustain liquid water on its surface (its equilibrium temperature is about $\sim$595\,K, suggesting a Venus-like atmosphere), spectroscopic metrics based on the capabilities of the forthcoming James Webb Space Telescope and the fact that the host star is rather inactive indicate that this is one of the most favorable known rocky exoplanets for atmospheric characterization.Comment: 14 pages, 10 figures, 6 tables, accepted for publication in A&

Three Warm Jupiters around Solar-analog Stars Detected with TESS*

We report the discovery and characterization of three giant exoplanets orbiting solar-analog stars, detected by the TESS space mission and confirmed through ground-based photometry and radial velocity measurements taken at La Silla observatory with FEROS. TOI-2373 b is a warm Jupiter orbiting its host star every ∼13.3 days, and is one of the most massive known exoplanet with a precisely determined mass and radius around a star similar to the Sun, with an estimated mass of m _p = ${9.3}_{-0.2}^{+0.2}\,{M}_{\mathrm{jup}}$ and a radius of r _p = ${0.93}_{-0.2}^{+0.2}\,{R}_{\mathrm{jup}}$ . With a mean density of $\rho ={14.4}_{-1.0}^{+0.9}\,{\rm{g}}\,{\mathrm{cm}}^{-3}$ , TOI-2373 b is among the densest planets discovered so far. TOI-2416 b orbits its host star on a moderately eccentric orbit with a period of ∼8.3 days and an eccentricity of e = ${0.32}_{-0.02}^{+0.02}$ . TOI-2416 b is more massive than Jupiter with m _p = ${3.0}_{-0.09}^{+0.10}\,{M}_{\mathrm{jup}}$ , however is significantly smaller with a radius of r _p = ${0.88}_{-0.02}^{+0.02},{R}_{\mathrm{jup}}$ , leading to a high mean density of $\rho ={5.4}_{-0.3}^{+0.3}\,{\rm{g}}\,{\mathrm{cm}}^{-3}$ . TOI-2524 b is a warm Jupiter near the hot Jupiter transition region, orbiting its star every ∼7.2 days on a circular orbit. It is less massive than Jupiter with a mass of m _p = ${0.64}_{-0.04}^{+0.04}\,{M}_{\mathrm{jup}}$ , and is consistent with an inflated radius of r _p = ${1.00}_{-0.03}^{+0.02}\,{R}_{\mathrm{jup}}$ , leading to a low mean density of $\rho ={0.79}_{-0.08}^{+0.08}\,{\rm{g}}\,{\mathrm{cm}}^{-3}$ . The newly discovered exoplanets TOI-2373 b, TOI-2416 b, and TOI-2524 b have estimated equilibrium temperatures of ${860}_{-10}^{+10}$ K, ${1080}_{-10}^{+10}$ K, and ${1100}_{-20}^{+20}$ K, respectively, placing them in the sparsely populated transition zone between hot and warm Jupiters