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In this paper, we address the problem of antimicrobial resistance in the case of Helicobacter pylori with a crystal engineering approach. Two antibiotics of the fluoroquinolone class, namely, levofloxacin (LEV) and ciprofloxacin (CIP), have been co-crystallized with the flavonoids quercetin (QUE), myricetin (MYR), and hesperetin (HES), resulting in the formation of four co-crystals, namely, LEV∙QUE, LEV∙MYR, LEV2∙HES, and CIP∙QUE. The co-crystals were obtained from solution, slurry, or mechanochemical mixing of the reactants. LEV∙QUE and LEV∙MYR were initially obtained as the ethanol solvates LEV∙QUE∙xEtOH and LEV∙MYR∙xEtOH, respectively, which upon thermal treatment yielded the unsolvated forms. All co-crystals were characterized by powder X-ray diffraction and thermal gravimetric analysis. The antibacterial performance of the four co-crystals LEV∙QUE, LEV∙MYR, LEV2∙HES, and CIP∙QUE in comparison with that of the physical mixtures of the separate components was tested via evaluation of the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). The results obtained indicate that the association with the co-formers, whether co-crystallized or forming a physical mixture with the active pharmaceutical ingredients (API), enhances the antimicrobial activity of the fluoroquinolones, allowing them to significantly reduce the amount of API otherwise required to display the same activity against H. pylori.

Background: Cytomegalovirus (CMV) is the most common agent of congenital infection in humans. It is a main cause of neurodevelopmental delay and sensorineural hearing loss in infancy. Since the 2000s, a number of studies have used Valganciclovir as a therapy for children with congenital CMV infection. Methods: In order to evaluate the efficacy of Valganciclovir in preventing clinical sequelae and its possible side effects, we performed a review of the published literature. This search was completed via PubMed for manuscripts published from January 2007 to December 2021, combining the MeSH words “Valganciclovir”, “Congenital”, and “Cytomegalovirus”. Results: A total of 27 articles were included (12 retrospective studies, 4 prospective studies, 1 randomized controlled trial, and 10 case reports). The clinical features were similar to those already described in the literature. The therapeutic protocols used were very different between the various studies included and neonatal antiviral treatments were only moderately effective. The therapy proved to be well-tolerated. Conclusions: The quality of the included studies and the sample size were limited due to the rarity of the disease. The use of different therapeutic protocols in terms of starting dates, doses, and durations made it impossible to compare and correctly evaluate the efficacy of the treatments. Randomized controlled trials are needed to establish the correct effective dose with the fewest side effects and the most efficient duration of therapy.

Bacterial pathogens employ a general strategy to overcome host defenses by coordinating the virulence gene expression using dedicated regulatory systems that could raise intricate networks. During the last twenty years, many studies of Helicobacter pylori, a human pathogen responsible for various stomach diseases, have mainly focused on elucidating the mechanisms and functions of virulence factors. In parallel, numerous studies have focused on the molecular mechanisms that regulate gene transcription to attempt to understand the physiological changes of the bacterium during infection and adaptation to the environmental conditions it encounters. The number of regulatory proteins deduced from the genome sequence analyses responsible for the correct orchestration of gene transcription appears limited to 14 regulators and three sigma factors. Furthermore, evidence is accumulating for new and complex circuits regulating gene transcription and H. pylori virulence. Here, we focus on the molecular mechanisms used by H. pylori to control gene transcription as a function of the principal environmental changes.

A Messinian and lithified horizon enriched in volcanic particles with thicknesses of 170–180 cm crops in the Camporotondo (CR) section (Marche, Italy). This volcanic-rich layer (VRL) was investigated by field plus mesoscopic observations, X-ray powder diffraction (XRPD), scanning electron microscopy (SEM), bulk composition methods and electron-microprobe analysis (EMPA). The quantitative textural features of volcanic and sedimentary components were determined by 2D image analysis. The lowermost massive 70–80 cm portion is free of sedimentary structure or characterised only by plane-parallel ones, whereas the uppermost one is undulated and cross-laminated. The XRPD and SEM outcomes unveil that the VRL of CR is mainly composed of glassy shards (≥80 area%), a variable amount of sedimentary minerals (2O present in the glass fraction suggest a provenance from a convergent geodynamic setting. The 2D image analysis on SEM observations show that the VRL-5.5 of CR is composed of very fine and sorted (averages of MZ of 5,72 and σi of 0,70), scarcely vesicular, glass shards, with similar long and short size dimensions, shape and roundness. The VRL-5.5 of CR is free of large minerals and fossils. The coupling of mesoscopic and microscopic determinations indicates that the lowermost interval was deposited such as a primary tephra, i.e., fallout pyroclasts sinking in seawater. Instead, the uppermost interval derives from local, low-energy and sin-depositional remobilisation of the same VRL-5.5. The textural attributes of the volcanic fractions, the sedimentological features and the thickness of the VRL at CR correspond to the westward deposit of a still unknown eruption likely occurred at 5.5 Ma.

Neisseria meningitidis colonizes the nasopharynx of humans, and pathogenic strains can disseminate into the bloodstream, causing septicemia and meningitis. NHBA is a surface-exposed lipoprotein expressed by all N. meningitidis strains in different isoforms. Diverse roles have been reported for NHBA in heparin-mediated serum resistance, biofilm formation, and adherence to host tissues. We determined that temperature controls the expression of NHBA in all strains tested, with increased levels at 30–32 °C compared to 37 °C. Higher NHBA expression at lower temperatures was measurable both at mRNA and protein levels, resulting in higher surface exposure. Detailed molecular analysis indicated that multiple molecular mechanisms are responsible for the thermoregulated NHBA expression. The comparison of mRNA steady-state levels and half-lives at 30 °C and 37 °C demonstrated an increased mRNA stability/translatability at lower temperatures. Protein stability was also impacted, resulting in higher NHBA stability at lower temperatures. Ultimately, increased NHBA expression resulted in higher susceptibility to complement-mediated killing. We propose that NHBA regulation in response to temperature downshift might be physiologically relevant during transmission and the initial step(s) of interaction within the host nasopharynx. Together these data describe the importance of NHBA both as a virulence factor and as a vaccine antigen during neisserial colonization and invasion.

Since the discovery of penicillin in the first half of the last century, antibiotics have become the pillars of modern medicine for fighting bacterial infections. However, pathogens resistant to antibiotic treatment have increased in recent decades, and efforts to discover new antibiotics have decreased. As a result, it is becoming increasingly difficult to treat bacterial infections successfully, and we look forward to more significant efforts from both governments and the scientific community to research new antibacterial drugs. This perspective article highlights the high potential of bacterial transcriptional and posttranscriptional regulators as targets for developing new drugs. We highlight some recent advances in the search for new compounds that inhibit their biological activity and, as such, appear very promising for treating bacterial infections.

The heat-shock response, a universal protective mechanism consisting of a transcriptional reprogramming of the cellular transcriptome, results in the accumulation of proteins which counteract the deleterious effects of heat-stress on cellular polypeptides. To quickly respond to thermal stress and trigger the heat-shock response, bacteria rely on different mechanisms to detect temperature variations, which can involve nearly all classes of biological molecules. In Campylobacter jejuni the response to heat-shock is transcriptionally controlled by a regulatory circuit involving two repressors, HspR and HrcA. In the present work we show that the heat-shock repressor HrcA acts as an intrinsic protein thermometer. We report that a temperature upshift up to 42 °C negatively affects HrcA DNA-binding activity to a target promoter, a condition required for de-repression of regulated genes. Furthermore, we show that this impairment of HrcA binding at 42 °C is irreversible in vitro, as DNA-binding was still not restored by reversing the incubation temperature to 37 °C. On the other hand, we demonstrate that the DNA-binding activity of HspR, which controls, in combination with HrcA, the transcription of chaperones’ genes, is unaffected by heat-stress up to 45 °C, portraying this master repressor as a rather stable protein. Additionally, we show that HrcA binding activity is enhanced by the chaperonin GroE, upon direct protein–protein interaction. In conclusion, the results presented in this work establish HrcA as a novel example of intrinsic heat-sensing transcriptional regulator, whose DNA-binding activity is positively modulated by the GroE chaperonin.

HP1043 is an essential orphan response regulator of Helicobacter pylori orchestrating multiple crucial cellular processes. Classified as a member of the OmpR/PhoB family of two-component systems, HP1043 exhibits a highly degenerate receiver domain and evolved to function independently of phosphorylation. Here, we investigated the HP1043 binding mode to a target sequence in the hp1227 promoter (Php1227). Scanning mutagenesis of HP1043 DNA-binding domain and consensus sequence led to the identification of residues relevant for the interaction of the protein with a target DNA. These determinants were used as restraints to guide a data-driven protein-DNA docking. Results suggested that, differently from most other response regulators of the same family, HP1043 binds in a head-to-head conformation to the Php1227 target promoter. HP1043 interacts with DNA largely through charged residues and contacts with both major and minor grooves of the DNA are required for a stable binding. Computational alanine scanning on molecular dynamics trajectory was performed to corroborate our findings. Additionally, in vitro transcription assays confirmed that HP1043 positively stimulates the activity of RNA polymerase.

In July and August 2019, two paroxysmal eruptions dramatically changed the morphology of the crater terrace that hosts the active vents of Stromboli volcano (Italy). Here, we document these morphological changes, by using 2259 UAS-derived photographs from eight surveys and Structure-from-Motion (SfM) photogrammetric techniques, resulting in 3D point clouds, orthomosaics, and digital surface models (DSMs) with resolution ranging from 8.1 to 12.4 cm/pixel. We focus on the morphological evolution of volcanic features and volume changes in the crater terrace and the upper part of the underlying slope (Sciara del Fuoco). We identify both crater terrace and lava field variations, with vents shifting up to 47 m and the accumulation of tephra deposits. The maximum elevation changes related to the two paroxysmal eruptions (in between May and September 2019) range from +41.4 to −26.4 m at the lava field and N crater area, respectively. Throughout September 2018–June 2020, the total volume change in the surveyed area was +447,335 m3. Despite Stromboli being one of the best-studied volcanoes worldwide, the UAS-based photogrammetry products of this study provide unprecedented high spatiotemporal resolution observations of its entire summit area, in a period when volcanic activity made the classic field inspections and helicopter overflights too risky. Routinely applied UAS operations represent an effective and evolving tool for volcanic hazard assessment and to support decision-makers involved in volcanic surveillance and civil protection operations.

The heat-shock response is defined by the transient gene-expression program that leads to the rapid accumulation of heat-shock proteins. This evolutionary conserved response aims at the preservation of the intracellular environment and represents a crucial pathway during the establishment of host–pathogen interaction. In the food-borne pathogen Campylobacter jejuni two transcriptional repressors, named HspR and HrcA, are involved in the regulation of the major heat-shock genes. However, the molecular mechanism underpinning HspR and HrcA regulatory function has not been defined yet. In the present work, we assayed and mapped the HspR and HrcA interactions on heat-shock promoters by high-resolution DNase I footprintings, defining their regulatory circuit, which governs C. jejuni heat-shock response. We found that, while DNA-binding of HrcA covers a compact region enclosing a single inverted repeat similar to the so-called Controlling Inverted Repeat of Chaperone Expression (CIRCE) sequence, HspR interacts with multiple high- and low-affinity binding sites, which contain HspR Associated Inverted Repeat (HAIR)-like sequences. We also explored the DNA-binding properties of the two repressors competitively on their common targets and observed, for the first time, that HrcA and HspR can directly interact and their binding on co-regulated promoters occurs in a cooperative manner. This mutual cooperative mechanism of DNA binding could explain the synergic repressive effect of HspR and HrcA observed in vivo on co-regulated promoters. Peculiarities of the molecular mechanisms exerted by HspR and HrcA in C. jejuni are compared to the closely related bacterium H. pylori that uses homologues of the two regulators.

Understanding the viscosity of mantle-derived magmas is needed to model their migration mechanisms and ascent rate from the source rock to the surface. High pressure−temperature experimental data are now available on the viscosity of synthetic melts, pure carbonatitic to carbonate−silicate compositions, anhydrous basalts, dacites and rhyolites. However, the viscosity of volatile-bearing melilititic melts, among the most plausible carriers of deep carbon, has not been investigated. In this study, we experimentally determined the viscosity of synthetic liquids with ~31 and ~39 wt% SiO2, 1.60 and 1.42 wt% CO2 and 5.7 and 1 wt% H2O, respectively, at pressures from 1 to 4.7 GPa and temperatures between 1265 and 1755 °C, using the falling-sphere technique combined with in situ X-ray radiography. Our results show viscosities between 0.1044 and 2.1221 Pa·s, with a clear dependence on temperature and SiO2 content. The atomic structure of both melt compositions was also determined at high pressure and temperature, using in situ multi-angle energy-dispersive X-ray diffraction supported by ex situ microFTIR and microRaman spectroscopic measurements. Our results yield evidence that the T−T and T−O (T = Si,Al) interatomic distances of ultrabasic melts are higher than those for basaltic melts known from similar recent studies. Based on our experimental data, melilititic melts are expected to migrate at a rate ~from 2 to 57 km·yr−1 in the present-day or the Archaean mantle, respectively.

The medically important human pathogen Helicobacter pylori relies on a collection of highly conserved heat-shock and chaperone proteins to preserve the integrity of cellular polypeptides and to control their homeostasis in response to external stress and changing environmental conditions. Among this set of chaperones, the CbpA protein has been shown to play a regulatory role in heat-shock gene regulation by directly interacting with the master stress-responsive repressor HspR. Apart from this regulatory role, little is known so far about CbpA functional activities. Using biochemistry and molecular biology approaches, we have started the in vitro functional characterization of H. pylori CbpA. Specifically, we show that CbpA is a multifunctional protein, being able to bind DNA and to stimulate the ATPase activity of the major chaperone DnaK. In addition, we report a preliminary observation suggesting that CbpA DNA-binding activity can be affected by the direct interaction with the heat-shock master repressor HspR, supporting the hypothesis of a reciprocal crosstalk between these two proteins. Thus, our work defines novel functions for H. pylori CbpA and stimulates further studies aimed at the comprehension of the complex regulatory interplay among chaperones and heat-shock transcriptional regulators.

Bacteria respond to different environmental stresses by reprogramming the transcription of specific genes whose proper expression is critical for their survival. In this regard, the heat-shock response, a widespread protective mechanism, triggers a sudden increase in the cellular concentration of different proteins, including molecular chaperones and proteases, to preserve protein folding and maintain cellular homeostasis. In the medically important gastric pathogen Helicobacter pylori the regulation of the principal heat-shock genes is under the transcriptional control of two repressor proteins named HspR and HrcA. To define the HrcA regulon, we carried out whole transcriptome analysis through RNA-sequencing, comparing the transcriptome of the H. pylori G27 wild type strain to that of the isogenic hrcA-knockout strain. Overall, differential gene expression analysis outlined 49 genes to be deregulated upon hrcA gene inactivation. Interestingly, besides controlling the transcription of genes coding for molecular chaperones and stress-related mediators, HrcA is involved in regulating the expression of proteins whose function is linked to several cellular processes crucial for bacterial survival and virulence. These include cell motility, membrane transporters, Lipopolysaccharide modifiers and adhesins. The role of HrcA as a central regulator of H. pylori transcriptome, as well as its interconnections with the HspR regulon are here analyzed and discussed. As the HrcA protein acts as a pleiotropic regulator, influencing the expression of several stress-unrelated genes, it may be considered a promising target for the design of new antimicrobial strategies.

This study documents the compositional variations of phenocrysts from a basaltic trachyandesitic sill emplaced in the Valle del Bove at Mt. Etna volcano (Sicily, Italy). The physicochemical conditions driving the crystallization and emplacement of the sill magma have been reconstructed by barometers, oxygen barometers, thermometers and hygrometers based on clinopyroxene, feldspar (plagioclase + K-feldspar) and titanomagnetite. Clinopyroxene is the liquidus phase, recording decompression and cooling paths decreasing from 200 to 0.1 MPa and from 1050 to 940 °C, respectively. Plagioclase and K-feldspar cosaturate the melt in a lower temperature interval of ~1000⁻870 °C. Cation exchanges in clinopyroxene (Mg-Fe) and feldspar (Ca-Na) indicate that magma ascent is accompanied by progressive H2O exsolution (up to ~2.2 wt. %) under more oxidizing conditions (up to ΔNNO + 0.5). Geospeedometric constraints provided by Ti⁻Al⁻Mg cation redistributions in titanomagnetite indicate that the travel time (up to 23 h) and ascent velocity of magma (up to 0.78 m/s) are consistent with those inferred for other eruptions at Mt. Etna. These kinetic effects are ascribed to a degassing-induced undercooling path caused principally by H2O loss at shallow crustal conditions. Rare earth element (REE) modeling based on the lattice strain theory supports the hypothesis that the sill magma formed from primitive basaltic compositions after clinopyroxene (≤41%) and plagioclase (≤12%) fractionation. Early formation of clinopyroxene at depth is the main controlling factor for the REE signature, whereas subsequent degassing at low pressure conditions enlarges the stability field of plagioclase causing trace element enrichments during eruption towards the surface.

All six minerals defined as “asbestos” by the existing regulation on asbestos hazard, i.e., actinolite, tremolite, anthophyllite, crocidolite and amosite amphiboles, and the serpentine-group mineral chrysotile are typical constituents of mafic and ultramafic magmatic rocks of ophiolitic sequences. However, little is known about the presence and distribution of naturally occurring asbestos (NOA) in plutonic felsic rocks. The Isadalu magmatic complex outcropping in central Sardinia and belonging to the post-variscan Permian volcanic cycle, is described here as an interesting occurrence of fibrous amphiboles in granitoid rocks. Field work and collected mineralogical/petrological data show that NOA fibers from the Isadalu complex belong compositionally to the actinolite-tremolite series. They were generated by metasomatic growth on pristine magmatic hornblende, at ca. 470 °C at 1 kbar, during sodic-calcic hydrothermal alteration. In terms of environmental hazard, the Isadalu complex represents a high-value case study, since the actinolite-bearing felsic rocks outcrop in a strongly anthropized area. Here, towns with local and regional strategic infrastructures (dams, pipes, hydroelectric power plants, water supply, roads) have been developed since the last century, also using the granitoid asbestos-rich stones. The aim of this study is to demonstrate that NOA and relative hazard are not univocally connected to a restricted typology of rocks. This result should be taken into account in any future work, procedure or regulation defining asbestos occurrences in natural environments.

Magma dynamics at Mt. Etna volcano are frequently recognized as the result of complex crystallization regimes that, at shallow crustal levels, unexpectedly change from H2O-undersaturated to H2O-saturated conditions, due to the impulsive and irregular arrival of volatile-rich magmas from mantle depths. On this basis, we have performed hydrous crystallization experiments for a quantitative understanding of the role of H2O in the differentiation of deep-seated trachybasaltic magmas at the key pressure of the Moho transition zone. For H2O = 2.1⁻3.2 wt %, the original trachybasaltic composition shifts towards phonotephritic magmas never erupted during the entire volcanic activity of Mt. Etna. Conversely, for H2O = 3.8⁻8.2 wt %, the obtained trachybasalts and basaltic trachyandesites reproduce most of the pre-historic and historic eruptions. The comparison with previous low pressure experimental data and natural compositions from Mt. Etna provides explanation for (1) the abundant release of H2O throughout the plumbing system of the volcano during impulsive ascent of deep-seated magmas; (2) the upward acceleration of magmas feeding gas-dominated, sustained explosive eruptions; (3) the physicochemical changes of gas-fluxed magmas ponding at shallow crustal levels; and (4) the huge gas emissions measured at the summit craters and flank vents which result in a persistent volcanic gas plume.

The ability to gauge the surroundings and modulate gene expression accordingly is a crucial feature for the survival bacterial pathogens. In this respect, the heat-shock response, a universally conserved mechanism of protection, allows bacterial cells to adapt rapidly to hostile conditions and to survive during environmental stresses. The important and widespread human pathogen Helicobacter pylori enrolls a collection of highly conserved heat-shock proteins to preserve cellular proteins and to maintain their homeostasis, allowing the pathogen to adapt and survive in the hostile niche of the human stomach. Moreover, various evidences suggest that some chaperones of H. pylori may play also non-canonical roles as, for example, in the interaction with the extracellular environment. In H. pylori, two dedicated transcriptional repressors, named HspR and HrcA, homologues to well-characterized regulators found in many other bacterial species, orchestrate the regulation of heat-shock proteins expression. Following twenty years of intense research, characterized by molecular, as well as genome-wide, approaches, it is nowadays possible to appreciate the complex picture representing the heat-shock regulation in H. pylori. Specifically, the HspR and HrcA repressors combine to control the transcription of target genes in a way that the HrcA regulon results embedded within the HspR regulon. Moreover, an additional level of control of heat-shock genes’ expression is exerted by a posttranscriptional feedback regulatory circuit in which chaperones interact and modulate HspR and HrcA DNA-binding activity. This review recapitulates our understanding of the roles and regulation of the most important heat-shock proteins of H. pylori, which represent a crucial virulence factor for bacterial infection and persistence in the human host.