Molecular insights into RmcA-mediated c-di-GMP consumption: Linking redox potential to biofilm morphogenesis in Pseudomonas aeruginosa

The ability of many bacteria to form biofilms contributes to their resilience and makes infections more difficult to treat. Biofilm growth leads to the formation of internal oxygen gradients, creating hypoxic subzones where cellular reducing power accumulates, and metabolic activities can be limited. The pathogen Pseudomonas aeruginosa counteracts the redox imbalance in the hypoxic biofilm subzones by producing redox-active electron shuttles (phenazines) and by secreting extracellular matrix, leading to an increased surface area-to-volume ratio, which favors gas exchange. Matrix production is regulated by the second messenger bis-(3′,5′)-cyclic-dimeric-guanosine monophosphate (c-di-GMP) in response to different environmental cues. RmcA (Redox modulator of c-di-GMP) from P. aeruginosa is a multidomain phosphodiesterase (PDE) that modulates c-di-GMP levels in response to phenazine availability. RmcA can also sense the fermentable carbon source arginine via a periplasmic domain, which is linked via a transmembrane domain to four cytoplasmic Per-Arnt-Sim (PAS) domains followed by a diguanylate cyclase (DGC) and a PDE domain. The biochemical characterization of the cytoplasmic portion of RmcA reported in this work shows that the PAS domain adjacent to the catalytic domain tunes RmcA PDE activity in a redox-dependent manner, by differentially controlling protein conformation in response to FAD or FADH2. This redox-dependent mechanism likely links the redox state of phenazines (via FAD/FADH2 ratio) to matrix production as indicated by a hyperwrinkling phenotype in a macrocolony biofilm assay. This study provides insights into the role of RmcA in transducing cellular redox information into a structural response of the biofilm at the population level. Conditions of resource (i.e. oxygen and nutrient) limitation arise during chronic infection, affecting the cellular redox state and promoting antibiotic tolerance. An understanding of the molecular linkages between condition sensing and biofilm structure is therefore of crucial importance from both biological and engineering standpoints.The authors would like to acknowledge Sapienza University of Rome [RM120172A7AD98EB to SR, RM1221815D52AB32 to APaiardini and AR12117A63EE6037; AR2221816C44C7B3 to CSR] for financial support. AUC experiments have received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 101004806. We thank Patrick England of the Plateforme de Biophysique Moléculaire of the C2RT (Institut Pasteur) for fruitful discussion

Subvert KEM to Break DEM: Practical Algorithm-Substitution Attacks on Public-Key Encryption

Motivated by the currently widespread concern about mass surveillance of encrypted communications, Bellare \emph{et al.} introduced at CRYPTO 2014 the notion of Algorithm-Substitution Attack (ASA) where the legitimate encryption algorithm is replaced by a subverted one that aims to undetectably exfiltrate the secret key via ciphertexts. Practically implementable ASAs on various cryptographic primitives (Bellare \emph{et al.}, CRYPTO\u2714 \& ACM CCS\u2715; Ateniese \emph{et al.}, ACM CCS\u2715; Berndt and Liśkiewicz, ACM CCS\u2717) have been constructed and analyzed, leaking the secret key successfully. Nevertheless, in spite of much progress, the practical impact of ASAs (formulated originally for symmetric key cryptography) on public-key (PKE) encryption operations remains unclear, primarily since the encryption operation of PKE does not involve the secret key, and also previously known ASAs become relatively inefficient for leaking the plaintext due to the logarithmic upper bound of exfiltration rate (Berndt and Liśkiewicz, ACM CCS\u2717). In this work, we formulate a practical ASA on PKE encryption algorithm which, perhaps surprisingly, turns out to be much more efficient and robust than existing ones, showing that ASAs on PKE schemes are far more effective and dangerous than previously believed. We mainly target PKE of hybrid encryption which is the most prevalent way to employ PKE in the literature and in practice. The main strategy of our ASA is to subvert the underlying key encapsulation mechanism (KEM) so that the session key encapsulated could be efficiently extracted, which, in turn, breaks the data encapsulation mechanism (DEM) enabling us to learn the plaintext itself. Concretely, our non-black-box yet quite general attack enables recovering the plaintext from only two successive ciphertexts and minimally depends on a short state of previous internal randomness. A widely used class of KEMs is shown to be subvertible by our powerful attack. Our attack relies on a novel identification and formalization of certain properties that yield practical ASAs on KEMs. More broadly, it points at and may shed some light on exploring structural weaknesses of other ``composed cryptographic primitives,\u27\u27 which may make them susceptible to more dangerous ASAs with effectiveness that surpasses the known logarithmic upper bound (i.e., reviewing composition as an attack enabler)

Identity-based Encryption Tightly Secure under Chosen-ciphertext Attacks

We propose the first identity-based encryption (IBE) scheme that is (almost) tightly secure against chosen-ciphertext attacks. Our scheme is efficient, in the sense that its ciphertext overhead is only seven group elements, three group elements more than that of the state-of-the-art passively (almost) tightly secure IBE scheme. Our scheme is secure in a multi-challenge setting, i.e., in face of an arbitrary number of challenge ciphertexts. The security of our scheme is based upon the standard symmetric external Diffie-Hellman assumption in pairing-friendly groups, but we also consider (less efficient) generalizations under weaker assumptions

Association of kidney disease measures with risk of renal function worsening in patients with type 1 diabetes

Background: Albuminuria has been classically considered a marker of kidney damage progression in diabetic patients and it is routinely assessed to monitor kidney function. However, the role of a mild GFR reduction on the development of stage 653 CKD has been less explored in type 1 diabetes mellitus (T1DM) patients. Aim of the present study was to evaluate the prognostic role of kidney disease measures, namely albuminuria and reduced GFR, on the development of stage 653 CKD in a large cohort of patients affected by T1DM. Methods: A total of 4284 patients affected by T1DM followed-up at 76 diabetes centers participating to the Italian Association of Clinical Diabetologists (Associazione Medici Diabetologi, AMD) initiative constitutes the study population. Urinary albumin excretion (ACR) and estimated GFR (eGFR) were retrieved and analyzed. The incidence of stage 653 CKD (eGFR < 60 mL/min/1.73 m2) or eGFR reduction > 30% from baseline was evaluated. Results: The mean estimated GFR was 98 \ub1 17 mL/min/1.73m2 and the proportion of patients with albuminuria was 15.3% (n = 654) at baseline. About 8% (n = 337) of patients developed one of the two renal endpoints during the 4-year follow-up period. Age, albuminuria (micro or macro) and baseline eGFR < 90 ml/min/m2 were independent risk factors for stage 653 CKD and renal function worsening. When compared to patients with eGFR > 90 ml/min/1.73m2 and normoalbuminuria, those with albuminuria at baseline had a 1.69 greater risk of reaching stage 3 CKD, while patients with mild eGFR reduction (i.e. eGFR between 90 and 60 mL/min/1.73 m2) show a 3.81 greater risk that rose to 8.24 for those patients with albuminuria and mild eGFR reduction at baseline. Conclusions: Albuminuria and eGFR reduction represent independent risk factors for incident stage 653 CKD in T1DM patients. The simultaneous occurrence of reduced eGFR and albuminuria have a synergistic effect on renal function worsening

The Seduction of Thanatos : Gabriele D’Annunzio and the Decadent Death

Peer reviewe