The underappreciated diversity of bile acid modifications.

The repertoire of modifications to bile acids and related steroidal lipids by host and microbial metabolism remains incompletely characterized. To address this knowledge gap, we created a reusable resource of tandem mass spectrometry (MS/MS) spectra by filtering 1.2 billion publicly available MS/MS spectra for bile-acid-selective ion patterns. Thousands of modifications are distributed throughout animal and human bodies as well as microbial cultures. We employed this MS/MS library to identify polyamine bile amidates, prevalent in carnivores. They are present in humans, and their levels alter with a diet change from a Mediterranean to a typical American diet. This work highlights the existence of many more bile acid modifications than previously recognized and the value of leveraging public large-scale untargeted metabolomics data to discover metabolites. The availability of a modification-centric bile acid MS/MS library will inform future studies investigating bile acid roles in health and disease.

Plasmid-encoded toxin defence mediates mutualistic microbial interactions.

Gut environments harbour dense microbial ecosystems in which plasmids are widely distributed. Plasmids facilitate the exchange of genetic material among microorganisms while enabling the transfer of a diverse array of accessory functions. However, their precise impact on microbial community composition and function remains largely unexplored. Here we identify a prevalent bacterial toxin and a plasmid-encoded resistance mechanism that mediates the interaction between Lactobacilli and Enterococci. This plasmid is widespread across ecosystems, including the rumen and human gut microbiota. Biochemical characterization of the plasmid revealed a defence mechanism against reuterin, a toxin produced by various gut microbes, such as Limosilactobacillus reuteri. Using a targeted metabolomic approach, we find reuterin to be prevalent across rumen ecosystems with impacts on microbial community structure. Enterococcus strains carrying the protective plasmid were isolated and their interactions with L. reuteri, the toxin producer, were studied in vitro. Interestingly, we found that by conferring resistance against reuterin, the plasmid mediates metabolic exchange between the defending and the attacking microbial species, resulting in a beneficial relationship or mutualism. Hence, we reveal here an ecological role for a plasmid-coded defence system in mediating a beneficial interaction.

Mechanisms of plant cell wall surveillance in response to pathogens, cell wall-derived ligands and the effect of expansins to infection resistance or susceptibility.

Cell wall integrity is tightly regulated and maintained given that non-physiological modification of cell walls could render plants vulnerable to biotic and/or abiotic stresses. Expansins are plant cell wall-modifying proteins active during many developmental and physiological processes, but they can also be produced by bacteria and fungi during interaction with plant hosts. Cell wall alteration brought about by ectopic expression, overexpression, or exogenous addition of expansins from either eukaryote or prokaryote origin can in some instances provide resistance to pathogens, while in other cases plants become more susceptible to infection. In these circumstances altered cell wall mechanical properties might be directly responsible for pathogen resistance or susceptibility outcomes. Simultaneously, through membrane receptors for enzymatically released cell wall fragments or by sensing modified cell wall barrier properties, plants trigger intracellular signaling cascades inducing defense responses and reinforcement of the cell wall, contributing to various infection phenotypes, in which expansins might also be involved. Here, we review the plant immune response activated by cell wall surveillance mechanisms, cell wall fragments identified as responsible for immune responses, and expansin's roles in resistance and susceptibility of plants to pathogen attack.

Nanoscale resolution of microbial fiber degradation in action.

The lives of microbes unfold at the micron scale, and their molecular machineries operate at the nanoscale. Their study at these resolutions is key toward achieving a better understanding of their ecology. We focus on cellulose degradation of the canonical Clostridium thermocellum system to comprehend how microbes build and use their cellulosomal machinery at these nanometer scales. Degradation of cellulose, the most abundant organic polymer on Earth, is instrumental to the global carbon cycle. We reveal that bacterial cells form 'cellulosome capsules' driven by catalytic product-dependent dynamics, which can increase the rate of hydrolysis. Biosynthesis of this energetically costly machinery and cell growth are decoupled at the single-cell level, hinting at a division-of-labor strategy through phenotypic heterogeneity. This novel observation highlights intrapopulation interactions as key to understanding rates of fiber degradation.

Clostridium perfringens in the spine: A rare cause of post-surgical infection.

BACKGROUND: Post-surgical infections of the spine occur in from 0% to 18% of cases. Postoperative spine infections due to Clostridium Perfringens (CP) resulting in necrotizing fasciitis are extremely rare. However, since they may be fatal, early and definitive treatment is critical. CASE DESCRIPTION A: 62-year-old male with a T8-T9 Type C fracture, in ASIA Grade "E" (neurologically intact) underwent a posterior T6-T10 arthrodesis. However, 2 weeks postoperatively, he developed a postoperative thoracic wound infection; the cultures were positive for CP. As the patient developed necrotizing fasciitis, emergent debridement, negative pressure continued drainage, and initiation of appropriate antibiotic therapy were critical. CONCLUSION: Postoperative spinal infections due to CP with accompanying necrotizing fasciitis are extremely rare. As these infections may be fatal, they must be rapidly diagnosed and treated.

Pulmonary cement embolism following transpedicular screws placement for thoracolumbar fractures.

BACKGROUND: Symptomatic pulmonary cement embolism in patients undergoing thoracic transpedicular fenestrated screw placement is rare. Here, we have added a 64-year-old female undergoing transpedicular screw placement for a T11 fracture who developed a pulmonary cement embolism intraoperatively and add this case to 13 others identified in the literature. CASE DESCRIPTION: A 64-year-old female presented with a type "C", ASIA "E" T11 fracture. The thoracolumbar pedicle screw fixation was supplemented with bone cement due to her underlying severe osteoporosis. During the fluoroscopy-guided supplementation with bone cement, a leak through the paravertebral venous system was noted. Thirty minutes later, the patient acutely developed extreme respiratory failure and required mechanical ventilation for the next 2 days. The diagnosis of pulmonary embolism due to bone cement was confirmed on a contrast computed tomography study of the chest. CONCLUSION: Symptomatic pulmonary cement embolization supplementing transpedicular screws placement for osteoporotic bone is rare. Here, we present a 64-year-old female who during transpedicular fixation of a T11 fracture developed an acute pulmonary embolism from the bone cement resulting in the need for 2 days of postoperative artificial ventilation.

Expansin-related proteins: biology, microbe-plant interactions and associated plant-defense responses.

Expansins, cerato-platanins and swollenins (which we will henceforth refer to as expansin-related proteins) are a group of microbial proteins involved in microbe-plant interactions. Although they share very low sequence similarity, some of their composing domains are near-identical at the structural level. Expansin-related proteins have their target in the plant cell wall, in which they act through a non-enzymatic, but still uncharacterized, mechanism. In most cases, mutagenesis of expansin-related genes affects plant colonization or plant pathogenesis of different bacterial and fungal species, and thus, in many cases they are considered virulence factors. Additionally, plant treatment with expansin-related proteins activate several plant defenses resulting in the priming and protection towards subsequent pathogen encounters. Plant-defence responses induced by these proteins are reminiscent of pattern-triggered immunity or hypersensitive response in some cases. Plant immunity to expansin-related proteins could be caused by the following: (i) protein detection by specific host-cell receptors, (ii) alterations to the cell-wall-barrier properties sensed by the host, (iii) displacement of cell-wall polysaccharides detected by the host. Expansin-related proteins may also target polysaccharides on the wall of the microbes that produced them under certain physiological instances. Here, we review biochemical, evolutionary and biological aspects of these relatively understudied proteins and different immune responses they induce in plant hosts.

Expansin-like Exl1 from Pectobacterium is a virulence factor required for host infection, and induces a defence plant response involving ROS, and jasmonate, ethylene and salicylic acid signalling pathways in Arabidopsis thaliana.

Expansins are encoded by some phytopathogenic bacteria and evidence indicates that they act as virulence factors for host infection. Here we analysed the expression of exl1 by Pectobacterium brasiliense and Pectobacterium atrosepticum. In both, exl1 gene appears to be under quorum sensing control, and protein Exl1 can be observed in culture medium and during plant infection. Expression of exl1 correlates with pathogen virulence, where symptoms are reduced in a Δexl1 mutant strain of P. atrosepticum. As well as Δexl1 exhibiting less maceration of potato plants, fewer bacteria are observed at distance from the inoculation site. However, bacteria infiltrated into the plant tissue are as virulent as the wild type, suggesting that this is due to alterations in the initial invasion of the tissue. Additionally, swarming from colonies grown on MacConkey soft agar was delayed in the mutant in comparison to the wild type. We found that Exl1 acts on the plant tissue, probably by remodelling of a cell wall component or altering the barrier properties of the cell wall inducing a plant defence response, which results in the production of ROS and the induction of marker genes of the JA, ET and SA signalling pathways in Arabidopsis thaliana. Exl1 inactive mutants fail to trigger such responses. This defence response is protective against Pectobacterium brasiliense and Botrytis cinerea in more than one plant species.

Analysis of two Mexican Pectobacterium brasiliense strains reveals an inverted relationship between c-di-GMP levels with exopolysaccharide production and swarming motility.

Pectobacterium is a diverse genus of phytopathogenic species from soil and water that cause infection either to restricted or multiple plant hosts. Phylogenetic analysis and metabolic fingerprinting of large numbers of genomes have expanded classification of Pectobacterium members. Pectobacterium brasiliense sp. nov has been elevated to the species level having detached from P. carotovorum. Here we present two P. brasiliense strains BF20 and BF45 isolated in Mexico from Opuntia and tobacco, respectively, which cluster into two different groups in whole genome comparisons with other Pectobacterium. We found that BF20 and BF45 strains are phenotypically different as BF45 showed more severe and rapid symptoms in comparison to BF20 in the host models celery and broccoli. Both strains produced similar levels of the main autoinducers, but BF45 shows an additional low abundant autoinducer compared to strain BF20. The two strains had different levels of c-di-GMP, which regulates the transition from motile to sessile lifestyle. In contrast to BF45, BF20 had the highest levels of c-di-GMP, was more motile (swarming), non-flocculant and less proficient in biofilm formation and exopolysaccharide production. Genomic comparisons revealed that differences in c-di-GMP accumulation and perhaps the associated phenotypes might be due to unique c-di-GMP metabolic genes in these two strains. Our results improve our understanding of the associations between phenotype and genotype and how this has shaped the physiology of Pectobacterium strains.

All-around management of a fungal isolate obtained from cheese spoilage as an environmental source: Direct approach from an undergrad student to a biotechnological characterization.

In this work, we present the results of an undergrad student from the perspective of its first approach as a principal researcher in a project. In order to gain practical experience, one of the options for students that have interest in pursuing a postgraduate program corresponds to a research stay in a laboratory of their selected field conducting a project for a period of 6 months. In this particular project, a fungal sample was isolated from Parmesan cheese spoilage and its enzymatic activity evaluated. Using simple and standardized protocols, the student was capable of identifying a possible biotechnological application for the isolate by detecting and categorizing the lipolytic activity. Through microculture characterization in potato dextrose agar (PDA) the genus of the sample was determined as Penicillium and confirmed by molecular analysis of the ITS1 and ITS4 regions. In order to examine comprehensively the potential of the new isolate, the extracellular and intracellular enzymatic activities were analyzed as well as four methods of cell rupture. From these results, sonication was determined as the best technique with 211 U/L as a maximum lipolytic value. To finalize the evaluation of the sample, the student determined the optimal pH and temperature as well as the thermotolerance of the crude extract obtaining a residual activity of 13% after 60 minutes of incubation at 45 °C. Upon conclusion of the research we could recognize that through a direct characterization of a fungal isolate using techniques that are widely known, the student was capable of determining and value one of the most interesting capabilities fungi has to offer; enzymatic activity, and that the knowledge obtained from established protocols enables and encourages the students to correlate the source from where they were obtained with potential biotechnological applications. © 2019 International Union of Biochemistry and Molecular Biology, 47(6):681-688, 2019.

Analysis of the Binding of Expansin Exl1, from Pectobacterium carotovorum, to Plant Xylem and Comparison to EXLX1 from Bacillus subtilis.

The plant xylem is a preferred niche for some important bacterial phytopathogens, some of them encoding expansin proteins, which bind plant cell walls. Yet, the identity of the substrate for bacterial expansins within the plant cell wall and the nature of its interaction with it are poorly known. Here, we determined the localization of two bacterial expansins with differing isoelectric points (and with differing binding patterns to cell wall extracts) on plant tissue through in vitro fluorophore labeling and confocal imaging. Differential localization was observed, in which Exl1 from Pectobacterium carotovorum located into the intercellular spaces between xylem vessels and adjacent cells of the plant xylem, whereas EXLX1 from Bacillus subtilis bound cell walls of most cell types. In isolated vascular tissue, however, both PcExl1 and BsEXLX1 preferentially bound to tracheary elements over the xylem fibers, even though both are composed of secondary cell walls. Fluorescence correlation spectroscopy, employed to analyze the interaction of expansins with isolated xylem, indicates that binding is governed by more than one factor, which could include interaction with more than one type of polymer in the fibers, such as cellulose and hemicellulose or pectin. Binding to different polysaccharides could explain the observed reduction of cellulolytic and xylanolytic activities in the presence of expansin, possibly because of competition for the substrate. Our findings are relevant for the comprehensive understanding of the pathogenesis by P. carotovorum during xylem invasion, a process in which Exl1 might be involved.

Hematoma traumático de vulva poco frecuente / Uncommon traumatic vulvar hematoma

El hematoma vulvar es la colección de sangre a nivel vulvar. Generalmente es secundario a un trauma obstétrico, lesiones por contusión o relaciones sexuales consensuadas. Se presenta una paciente de 21 años de edad que acude a esta unidad de salud en el mes de enero de 2017, refiriendo que posterior a un acto sexual comenzó a presentar aumento progresivo de volumen de vulva, acompañado de dolor y de dificultad para la deambulación. Se realizó incisión y drenaje del hematoma, se dan puntos hemostáticos, se colocó drenaje y se inició terapia antimicrobiana con ceftriaxone y metronidazol. El hematoma vulvar de la magnitud presentada constituyó una urgencia quirúrgica por el compromiso de la funcionalidad del aparato genitourinario. El drenaje del hematoma y la indicación de antimicrobianos permitió su evolución satisfactoria(AU)

The vulvar hematoma is the collection of blood in the vulvar area. It is usually secondary to obstetric trauma, contusion injuries or consensual sexual relations. We present a 21-year-old patient who came to this health unit in January 2017, reporting that after a sexual intercourse, she began to present a progressive increase in vulvar volume, pain and difficulty in walking. The hematoma was incised and drained, hemostatic stitches were given, drainage was placed and ceftriaxone and metronidazole were the antimicrobial therapy of choice. The vulvar hematoma of this magnitude was a surgical emergency due to the compromised functionality of the genitourinary system. The drainage of the hematoma and the indication of antimicrobials allowed satisfactory evolution(AU)

Schizophyllum commune: An unexploited source for lignocellulose degrading enzymes.

Lignocellulose represents the most abundant source of carbon in the Earth. Thus, fraction technology of the biomass turns up as an emerging technology for the development of biorefineries. Saccharification and fermentation processes require the formulation of enzymatic cocktails or the development of microorganisms (naturally or genetically modified) with the appropriate toolbox to produce a cost-effective fermentation technology. Therefore, the search for microorganisms capable of developing effective cellulose hydrolysis represents one of the main challenges in this era. Schizophyllum commune is an edible agarical with a great capability to secrete a myriad of hydrolytic enzymes such as xylanases and endoglucanases that are expressed in a high range of substrates. In addition, a large number of protein-coding genes for glycoside hydrolases, oxidoreductases like laccases (Lacs; EC 1.10.3.2), as well as some sequences encoding for lytic polysaccharide monooxygenases (LPMOs) and expansins-like proteins demonstrate the potential of this fungus to be applied in different biotechnological process. In this review, we focus on the enzymatic toolbox of S. commune at the genetic, transcriptomic, and proteomic level, as well as the requirements to be employed for fermentable sugars production in biorefineries. At the end the trend of its use in patent registration is also reviewed.

Simple screening protocol for identification of potential mycoremediation tools for the elimination of polycyclic aromatic hydrocarbons and phenols from hyperalkalophile industrial effluents.

A number of fungal strains belonging to the ascomycota, basidiomycota and zygomycota genera were subjected to an in vitro screening regime to assess their ligninolytic activity potential, with a view to their potential use in mycoremediation-based strategies to remove phenolic compounds and polycyclic aromatic hydrocarbons (PAHs) from industrial wastewaters. All six basidiomycetes completely decolorized remazol brilliant blue R (RBBR), while also testing positive in both the guaiacol and gallic acid tests indicating good levels of lignolytic activity. All the fungi were capable of tolerating phenanthrene, benzo-α- pyrene, phenol and p-chlorophenol in agar medium at levels of 10 ppm. Six of the fungal strains, Pseudogymnoascus sp., Aspergillus caesiellus, Trametes hirsuta IBB 450, Phanerochate chrysosporium ATCC 787, Pleurotus ostreatus MTCC 1804 and Cadophora sp. produced both laccase and Mn peroxidase activity in the ranges of 200-560 U/L and 6-152 U/L, respectively, in liquid media under nitrogen limiting conditions. The levels of adsorption of the phenolic and PAHs were negligible with 99% biodegradation being observed in the case of benzo-α-pyrene, phenol and p-chlorophenol. The aforementioned six fungal strains were also found to be able to effectively treat highly alkaline industrial wastewater (pH 12.4). When this wastewater was supplemented with 0.1 mM glucose, all of the tested fungi, apart from A. caesiellus, displayed the capacity to remove both the phenolic and PAH compounds. Based on their biodegradative capacity we found T. hirsuta IBB 450 and Pseudogymnoascus sp., to have the greatest potential for further use in mycoremediation based strategies to treat wastestreams containing phenolics and PAHs.

Characterization of lignocellulolytic activities from fungi isolated from the deep-sea sponge Stelletta normani.

Extreme habitats have usually been regarded as a source of microorganisms that possess robust proteins that help enable them to survive in such harsh conditions. The deep sea can be considered an extreme habitat due to low temperatures (<5°C) and high pressure, however marine sponges survive in these habitats. While bacteria derived from deep-sea marine sponges have been studied, much less information is available on fungal biodiversity associated with these sponges. Following screening of fourteen fungi isolated from the deep-sea sponge Stelletta normani sampled at a depth of 751 metres, three halotolerant strains (TS2, TS11 and TS12) were identified which displayed high CMCase and xylanase activities. Molecular based taxonomic approaches identified these strains as Cadophora sp. TS2, Emericellopsis sp. TS11 and Pseudogymnoascus sp. TS 12. These three fungi displayed psychrotolerance and halotolerant growth on CMC and xylan as sole carbon sources, with optimal growth rates at 20°C. They produced CMCase and xylanase activities, which displayed optimal temperature and pH values of between 50-70°C and pH 5-8 respectively, together with good thermostability and halotolerance. In solid-state fermentations TS2, TS11 and TS12 produced CMCases, xylanases and peroxidase/phenol oxidases when grown on corn stover and wheat straw. This is the first time that CMCase, xylanase and peroxidase/phenol oxidase activities have been reported in these three fungal genera isolated from a marine sponge. Given the biochemical characteristics of these ligninolytic enzymes it is likely that they may prove useful in future biomass conversion strategies involving lignocellulosic materials.

A Pycnoporus sanguineus laccase for denim bleaching and its comparison with an enzymatic commercial formulation.

A laccase from the basidiomycete Pycnoporus sanguineus strain RVAN5 was evaluated for its ability to decolorize synthetic dyes and denim bleaching. Dye color reduction and denim bleaching were monitored at different dye concentrations and incubation times. Dye decolorization by Pycnoporus sanguineus fungal crude extract (FCE) ranged from 80 to 96% within 2-4 h at 25-65 °C. Comparable results were obtained when violuric acid (VA) was added as mediator to the FCE, however, the number of decolorized dyes increased significantly. Dye decolorization rates with VA varied of initial and final optical density (595 nm) values of 2.5-3.0 and 0.2-0.02, respectively. P. sanguineus FCE had no substantial effect on denim bleaching when used alone, notwithstanding, the mixture of FCE with VA (10 mM) showed significant denim color reduction values and considerably higher than those obtained with a bleaching enzyme from a commercial formulation; CIElab values obtained with FCE/VA mixture were of ΔL = 6.4, versus a ΔL 1.4 value obtained with an enzyme from commercial formulation.

A novel expansin protein from the white-rot fungus Schizophyllum commune.

A novel expansin protein (ScExlx1) was found, cloned and expressed from the Basidiomycete fungus Schizophylum commune. This protein showed the canonical features of plant expansins. ScExlx1 showed the ability to form "bubbles" in cotton fibers, reduce the size of avicel particles and enhance reducing sugar liberation from cotton fibers pretreated with the protein and then treated with cellulases. ScExlx1 was able to bind cellulose, birchwood xylan and chitin and this property was not affected by different sodium chloride concentrations. A novel property of ScExlx1 is its capacity to enhance reducing sugars (N-acetyl glucosamine) liberation from pretreated chitin and further added with chitinase, which has not been reported for any expansin or expansin-like protein. To the best of our knowledge, this is the first report of a bona fide fungal expansin found in a basidiomycete and we could express the bioactive protein in Pichia pastoris.

Herida penetrante del cráneo / Skull penetrating wound

El traumatismo craneoencefálico es común en los servicios de urgencia de instituciones que atienden a pacientes politraumatizados y se ha convertido en un problema de salud para muchos países. El traumatismo penetrante del cráneo ocupa un lugar especial por su baja frecuencia. En este trabajo se presenta el caso de un paciente varón, de 52 años de edad, que sufrió una herida penetrante del cráneo producida por un arma blanca que quedó retenida en la región frontotemporal izquierda. Tras un estudio imaginológico se procedió al tratamiento quirúrgico de urgencia, y el paciente evoluciona satisfactoriamente después de 25 días de hospitalización. En la actualidad se encuentra en tratamiento de rehabilitación por una hemiparesia derecha residual(AU)

The cranioencephalic trauma is common in the emergence centers to care for patients with multiple traumata and it becames in a health problem in many countries. Skull penetrating trauma is located in a special place due to its low frequency. In present paper a case of male patient aged 52 severely skull-injured with penetrating wound caused by a cold steel that remained introduced into the left frontotemporal region. After an imaging study the emergence surgical treatment was applied and patient evolves adequately after 25 days of hospitalization. Nowadays, she is under rehabilitation treatment due to a residual right hemiparesis(AU)

Herida penetrante del cráneo / Skull penetrating wound

El traumatismo craneoencefálico es común en los servicios de urgencia de instituciones que atienden a pacientes politraumatizados y se ha convertido en un problema de salud para muchos países. El traumatismo penetrante del cráneo ocupa un lugar especial por su baja frecuencia. En este trabajo se presenta el caso de un paciente varón, de 52 años de edad, que sufrió una herida penetrante del cráneo producida por un arma blanca que quedó retenida en la región frontotemporal izquierda. Tras un estudio imaginológico se procedió al tratamiento quirúrgico de urgencia, y el paciente evoluciona satisfactoriamente después de 25 días de hospitalización. En la actualidad se encuentra en tratamiento de rehabilitación por una hemiparesia derecha residual(AU)

The cranioencephalic trauma is common in the emergence centers to care for patients with multiple traumata and it becames in a health problem in many countries. Skull penetrating trauma is located in a special place due to its low frequency. In present paper a case of male patient aged 52 severely skull-injured with penetrating wound caused by a cold steel that remained introduced into the left frontotemporal region. After an imaging study the emergence surgical treatment was applied and patient evolves adequately after 25 days of hospitalization. Nowadays, she is under rehabilitation treatment due to a residual right hemiparesis(AU)

Revisión por pares: ¿Qué es y para qué sirve? / Peer review: what it's and what it's for?

La revisión por pares es una herramienta usada en la valoración crítica de los manuscritos enviados por los expertos a las revistas científicas; su implementación genera dudas respecto al proceso de evaluación. Con el objetivo de aclarar el proceso de revisión por pares en este artículo se explican las interrogantes más frecuentes y se hace referencia a las ventajas, desventajas y a cómo mejorar el proceso, así como también su importancia, teniendo en cuenta que todas las investigaciones deberían ser sometidas a este proceso, para su validación previa a la publicación. Por otro lado, se comenta la situación actual en Latinoamérica, con el fin de establecer medidas que ayuden al mejoramiento del proceso. Finalmente, comentamos los parámetros actuales en relación con el proceso de revisión por pares y su metodología...

The peer review is a tool used in the critical valuation of the manuscripts sent to the magazines by the experts, which generate doubts as far as this process of evaluation; with the aim of clarifying the process of peer review the questions are explained but frequent and reference to the advantages becomes, disadvantages and like improving the process, as well as its importance, considering that all the investigations would have to be put under this process, for its previous validation to the publication. On the other hand, the present situation in Latin America comments, in order to establish measures that help the improvement of the process. Finally, we commented the present parameters in relation to the process of peer review and their methodology...