A bioengineered probiotic for the oral delivery of a peptide Kv1.3 channel blocker to treat rheumatoid arthritis.

Engineered microbes for the delivery of biologics are a promising avenue for the treatment of various conditions such as chronic inflammatory disorders and metabolic disease. In this study, we developed a genetically engineered probiotic delivery system that delivers a peptide to the intestinal tract with high efficacy. We constructed an inducible system in the probiotic Lactobacillus reuteri to secrete the Kv1.3 potassium blocker ShK-235 (LrS235). We show that LrS235 culture supernatants block Kv1.3 currents and preferentially inhibit human T effector memory (TEM) lymphocyte proliferation in vitro. A single oral gavage of healthy rats with LrS235 resulted in sufficient functional ShK-235 in the circulation to reduce inflammation in a delayed-type hypersensitivity model of atopic dermatitis mediated by TEM cells. Furthermore, the daily oral gavage of LrS235 dramatically reduced clinical signs of disease and joint inflammation in rats with a model of rheumatoid arthritis without eliciting immunogenicity against ShK-235. This work demonstrates the efficacy of using the probiotic L. reuteri as a novel oral delivery platform for the peptide ShK-235 and provides an efficacious strategy to deliver other biologics with great translational potential.

Early Biomarker Signatures in Surgical Sepsis.

INTRODUCTION: Sepsis has complex, time-sensitive pathophysiology and important phenotypic subgroups. The objective of this study was to use machine learning analyses of blood and urine biomarker profiles to elucidate the pathophysiologic signatures of subgroups of surgical sepsis patients. METHODS: This prospective cohort study included 243 surgical sepsis patients admitted to a quaternary care center between January 2015 and June 2017. We applied hierarchical clustering to clinical variables and 42 blood and urine biomarkers to identify phenotypic subgroups in a development cohort. Clinical characteristics and short-term and long-term outcomes were compared between clusters. A naïve Bayes classifier predicted cluster labels in a validation cohort. RESULTS: The development cohort contained one cluster characterized by early organ dysfunction (cluster I, n = 18) and one cluster characterized by recovery (cluster II, n = 139). Cluster I was associated with higher Acute Physiologic Assessment and Chronic Health Evaluation II (30 versus 16, P < 0.001) and SOFA scores (13 versus 5, P < 0.001), greater prevalence of chronic cardiovascular and renal disease (P < 0.001) and septic shock (78% versus 17%, P < 0.001). Cluster I had higher mortality within 14 d of sepsis onset (11% versus 1.5%, P = 0.001) and within 1 y (44% versus 20%, P = 0.032), and higher incidence of chronic critical illness (61% versus 30%, P = 0.001). The Bayes classifier achieved 95% accuracy and identified two clusters that were similar to development cohort clusters. CONCLUSIONS: Machine learning analyses of clinical and biomarker variables identified an early organ dysfunction sepsis phenotype characterized by inflammation, renal dysfunction, endotheliopathy, and immunosuppression, as well as poor short-term and long-term clinical outcomes.

Engineering bacterial thiosulfate and tetrathionate sensors for detecting gut inflammation.

There is a groundswell of interest in using genetically engineered sensor bacteria to study gut microbiota pathways, and diagnose or treat associated diseases. Here, we computationally identify the first biological thiosulfate sensor and an improved tetrathionate sensor, both two-component systems from marine Shewanella species, and validate them in laboratory Escherichia coli Then, we port these sensors into a gut-adapted probiotic E. coli strain, and develop a method based upon oral gavage and flow cytometry of colon and fecal samples to demonstrate that colon inflammation (colitis) activates the thiosulfate sensor in mice harboring native gut microbiota. Our thiosulfate sensor may have applications in bacterial diagnostics or therapeutics. Finally, our approach can be replicated for a wide range of bacterial sensors and should thus enable a new class of minimally invasive studies of gut microbiota pathways.

Genome alterations associated with improved transformation efficiency in Lactobacillus reuteri.

BACKGROUND: Lactic acid bacteria (LAB) are one of the microorganisms of choice for the development of protein delivery systems for therapeutic purposes. Although there are numerous tools to facilitate genome engineering of lactobacilli; transformation efficiency still limits the ability to engineer their genomes. While genetically manipulating Lactobacillus reuteri ATCC PTA 6475 (LR 6475), we noticed that after an initial transformation, several LR 6475 strains significantly improved their ability to take up plasmid DNA via electroporation. Our goal was to understand the molecular basis for how these strains acquired the ability to increase transformation efficiency. RESULTS: Strains generated after transformation of plasmids pJP067 and pJP042 increased their ability to transform plasmid DNA about one million fold for pJP067, 100-fold for pSIP411 and tenfold for pNZ8048. Upon sequencing of the whole genome from these strains, we identified several genomic mutations and rearrangements, with all strains containing mutations in the transformation related gene A (trgA). To evaluate the role of trgA in transformation of DNA, we generated a trgA null that improved the transformation efficiency of LR 6475 to transform pSIP411 and pJP067 by at least 100-fold, demonstrating that trgA significantly impairs the ability of LR 6475 to take-up plasmid DNA. We also identified genomic rearrangements located in and around two prophages inserted in the LR 6475 genome that included deletions, insertions and an inversion of 336 Kb. A second group of rearrangements was observed in a Type I restriction modification system, in which the specificity subunits underwent several rearrangements in the target recognition domain. Despite the magnitude of these rearrangements in the prophage genomes and restriction modification systems, none of these genomic changes impacted transformation efficiency to the level induced by trgA. CONCLUSIONS: Our findings demonstrate how genetic manipulation of LR 6475 with plasmid DNA leads to genomic changes that improve their ability to transform plasmid DNA; highlighting trgA as the primary driver of this phenotype. Additionally, this study also underlines the importance of characterizing genetic changes that take place after genome engineering of strains for therapeutic purposes.

Sex-specific differences in physiological parameters related to SARS-CoV-2 infections among a national cohort (COVI-GAPP study).

Considering sex as a biological variable in modern digital health solutions, we investigated sex-specific differences in the trajectory of four physiological parameters across a COVID-19 infection. A wearable medical device measured breathing rate, heart rate, heart rate variability, and wrist skin temperature in 1163 participants (mean age = 44.1 years, standard deviation [SD] = 5.6; 667 [57%] females). Participants reported daily symptoms and confounders in a complementary app. A machine learning algorithm retrospectively ingested daily biophysical parameters to detect COVID-19 infections. COVID-19 serology samples were collected from all participants at baseline and follow-up. We analysed potential sex-specific differences in physiology and antibody titres using multilevel modelling and t-tests. Over 1.5 million hours of physiological data were recorded. During the symptomatic period of infection, men demonstrated larger increases in skin temperature, breathing rate, and heart rate as well as larger decreases in heart rate variability than women. The COVID-19 infection detection algorithm performed similarly well for men and women. Our study belongs to the first research to provide evidence for differential physiological responses to COVID-19 between females and males, highlighting the potential of wearable technology to inform future precision medicine approaches.

Concentrations of Serum Brain Injury Biomarkers Following SARS-CoV-2 Infection in Individuals with and without Long-COVID-Results from the Prospective Population-Based COVI-GAPP Study.

It is unknown whether neurological symptoms are associated with brain injury after SARS-CoV-2 infections and whether brain injury and related symptoms also emerge in Long-COVID patients. Biomarkers such as serum neurofilament light chain (sNfL) and glial fibrillary acidic protein (sGFAP) can be used to elucidate neuro-axonal and astroglial injuries. We investigated whether these biomarkers are associated with COVID-19 infection status, associated symptoms and Long-COVID. From 146 individuals of the general population with a post-acute, mild-to-moderate SARS-CoV-2 infection, sNfL and sGFAP were measured before, during and after (five and ten months) the infection. Individual symptoms and Long-COVID status were assessed using questionnaires. Neurological associated symptoms were described for individuals after a mild and moderate COVID-19 infection; however, sNfL (p = 0.74) and sGFAP (p = 0.24) did not change and were not associated with headache (p = 0.51), fatigue (p = 0.93), anosmia (p = 0.77) or ageusia (p = 0.47). In Long-COVID patients, sGFAP (p = 0.038), but not sNfL (p = 0.58), significantly increased but was not associated with neurological associated symptoms. Long-COVID status, but not post-acute SARS-CoV-2 infections, may be associated with astroglial injury/activation, even if neurological associated symptoms were not correlated.

Challenges and Pitfalls in the Engineering of Human Interleukin 22 (hIL-22) Secreting Lactobacillus reuteri.

Engineered microbes for the delivery of intestinally directed therapeutics is a promising avenue for the treatment of various intestinal diseases including inflammatory bowel disease (IBD) and intestinal graft vs. host disease (GVHD). This modality of treatment would allow for the targeted delivery of therapeutics to the site of inflammation or disease while minimizing the systemic side effects that often accompany treatment of these pathologies. Here, we show the challenges encountered and overcome in successfully engineering Lactobacillus reuteri to secrete high levels of biologically active human interleukin 22 (hIL-22). Initial hIL-22 constructs secreted high levels of hIL-22, however we found the majority of hIL-22 was cleaved and not biologically active. Several strategies were explored to improve the production of intact hIL-22, with the optimization of the signal sequence for peptide secretion having the most impact of production of intact hIL-22. This resulted in L. reuteri secreting high concentrations (up to 700 ng/mL) of hIL-22. Bioactivity of hIL-22 was confirmed by the secretion of interleukin 10 (IL-10) from the colon cancer derived epithelial cell line Colo205 and the secretion of Regenerating islet-derived protein 3 alpha (Reg3α) from human jejunal enteroids. The secretion of bioactive hIL-22 imposed a significant cost for L. reuteri as bacterial growth was significantly impaired upon induction. Future challenges and optimization strategies for the delivery of hIL-22 to the human intestinal tract are discussed.

Discovery and Validation of Urinary Molecular Signature of Early Sepsis.

Identify alterations in gene expression unique to systemic and kidney-specific pathophysiologic processes using whole-genome analyses of RNA isolated from the urinary cells of sepsis patients. DESIGN: Prospective cohort study. SETTING: Quaternary care academic hospital. PATIENTS: A total of 266 sepsis and 82 control patients enrolled between January 2015 and February 2018. INTERVENTIONS: Whole-genome transcriptomic analysis of messenger RNA isolated from the urinary cells of sepsis patients within 12 hours of sepsis onset and from control subjects. MEASUREMENTS AND MAIN RESULTS: The differentially expressed probes that map to known genes were subjected to feature selection using multiple machine learning techniques to find the best subset of probes that differentiates sepsis from control subjects. Using differential expression augmented with machine learning ensembles, we identified a set of 239 genes in urine, which show excellent effectiveness in classifying septic patients from those with chronic systemic disease in both internal and independent external validation cohorts. Functional analysis indexes disrupted biological pathways in early sepsis and reveal key molecular networks driving its pathogenesis. CONCLUSIONS: We identified unique urinary gene expression profile in early sepsis. Future studies need to confirm whether this approach can complement blood transcriptomic approaches for sepsis diagnosis and prognostication.

A revision of Prespelea Park (Staphylinidae, Pselaphinae).

We revise the genus Prespelea Park, redefining and redescribing the two previously known species, P. copelandi Park and P. quirsfeldi Park, and adding ten new species: P. parki Caterino & Vásquez-Vélez, sp. n., P. minima Caterino & Vásquez-Vélez, sp. n., P. morsei Caterino & Vásquez-Vélez, sp. n., P. divergens Caterino & Vásquez-Vélez, sp. n., P. carltoni Caterino & Vásquez-Vélez, sp. n., P. myersae Caterino & Vásquez-Vélez, sp. n., P. georgiensis Caterino & Vásquez-Vélez, sp. n., P. enigma Caterino & Vásquez-Vélez, sp. n., P. wagneri Caterino & Vásquez-Vélez, sp. n., and P. basalis Caterino & Vásquez-Vélez, sp. n.. The genus is still only known from a relatively small area in the southern Appalachian Mountains, but the diversity is much greater than previously suspected. The new species exhibit considerable diversity in male secondary sexual characters. A preliminary phylogenetic analysis cannot conclusively resolve the polarity of eye and wing reduction across Speleobamini, but the monophyly of Park's subgenus Fusjugama, if expanded to include all species with full-eyed and winged males, is not supported, and we therefore synonymize it with Prespeleas. str.

Genetic Tools for the Enhancement of Probiotic Properties.

The Lactobacillus genus is a diverse group of microorganisms, many of which are of industrial and medical relevance. Several Lactobacillus species have been used as probiotics, organisms that when present in sufficient quantities confer a health benefit to the host. A significant limitation to the mechanistic understanding of how these microbes provide health benefits to their hosts and how they can be used as therapeutic delivery systems has been the lack of genetic strategies to efficiently manipulate their genomes. This article will review the development and employment of traditional genetic tools in lactobacilli and highlight the latest methodologies that are allowing for precision genome engineering of these probiotic organisms. The application of these tools will be key in providing mechanistic insights into probiotics as well as maximizing the value of lactobacilli as either a traditional probiotic or as a platform for the delivery of therapeutic proteins. Finally, we will discuss concepts that we consider relevant for the delivery of engineered therapeutics to the human gut.

Corrigenda: Caterino M, Vásquez-Vélez L (2017) A revision of Prespelea Park (Staphylinidae, Pselaphinae). ZooKeys 685: 105-130. https://doi.org/10.3897/zookeys.685.13811.

[This corrects the article DOI: 10.3897/zookeys.685.13811.].

Revision of the genus Pselaphomorphus Motschulsky, 1855 (Staphylinidae, Pselaphinae: Jubini).

The genus Pselaphomorphus Motschulsky, 1855 is revised to include 33 species. The type species, Pselaphomorphus sculpturatus Motschulsky, 1855, and the previously described species P. longiceps Raffray, 1890 and P. microphthalmus Raffray, 1890 are redescribed, and the original description of P. brevipennis Raffray, 1917 and P. bruchi Raffray, 1918 are translated from Latin and French to English. The following species are described as new: P. acutispinosus n. sp. from Peru, P. agametopus n. sp. from Colombia, P. azuayensis n. sp. from Ecuador, P. bolivarensis n. sp. from Venezuela, P. breviantennae n. sp. from Bolivia, P. carenobothrus n. sp. from Peru, P. carltoni n. sp. from Ecuador, P. cayennensis n. sp. from French Guiana, P. chandleri n. sp. from Panama, P. fluminosus n. sp. from Venezuela, P. iguazuensis n. sp. from Argentina, P. incorum n. sp. from Peru, P. longissimus n. sp. from Panama, P. macrommatus n. sp. from Ecuador, P. maldonadensis n. sp. from Peru, P. mayaorum n. sp. from Honduras, P. melanus n. sp. from Costa Rica, P. montsineryensis n. sp. from French Guiana, P. motschulskyi n. sp. from Costa Rica, P. newtoni n. sp. from Brazil, P. parki n. sp. from Costa Rica, P. raffrayi n. sp. from Peru, P. santaecrucis n. sp. from Bolivia, P. simplicipenis n. sp. from French Guiana, P. thayerae n. sp. from Brazil, P. tucumanensis n. sp. from Argentina, P. veracruzanus n. sp. and P. wagneri n. sp., both from Mexico. A key to the species, color photographs and genitalia illustrations are provided.

Revision of the genus Pselaphomorphus Motschulsky, 1855 (Staphylinidae, Pselaphinae: Jubini). Corrections.

In my revision of the genus Pselaphomorphus Motschulsky, 1855 (Vásquez-Vélez 2016) the depository of the holotype of P. cayennensis Vásquez-Vélez, 2016 was not listed. As a result, the requirements of Article 16.4.2 of the International Code of Zoological Nomenclature (International Commission on Zoological Nomenclature 1999) have not been met, making the name unavailable. This note aims at correcting the earlier mistake: the holotype of P. cayennensis is deposited at the Museum of natural history (Muséum d'histoire naturelle), Geneva, Switzerland.

Infarto renal y disección carotídea espontánea: complicaciones de la displasia fibromuscular / Spontaneous renal infarction and carotid dissection: complications of fibromuscular dysplasia

Resumen La displasia fibromuscular es una enfermedad no inflamatoria y no aterosclerótica, que puede afectar cualquier lecho arterial; sin embargo, las arterias renales y la carótida interna son las más comprometidas. Se presenta el caso de un paciente que cursó con dolor abdominal y cefalea hemicraneana posterior, con síndrome de Horner incompleto. En arteriografía renal y panangiografía cerebral realizada por Cardiología intervencionista se observó infarto renal derecho y disección de la carótida interna derecha con formación de pseudoaneurisma. Se hizo terapia endovascular con angioplastia y se dio de alta con antiagregación dual. Es importante conocer este tipo de presentación clínica ya que el diagnóstico podría confundirse con otras enfermedades que generan signos y síntomas similares; por consiguiente, es ideal tener sospecha clínica alta para evitar retrasos en el manejo.

Abstract Fibromuscular dysplasia is a non-inflammatory and non-atherosclerotic disease that can affect any arterial bed, with the renal and the internal carotid arteries being the most compromised. A case is presented on a patient that had abdominal pain and pain in the back of the head, and an incomplete Horner syndrome. In the renal arteriography and cerebral pan-angiography carried out by interventionist Cardiology, a right renal infarction and dissection of the right internal carotid was observed with a pseudo-aneurysm formation. Intravenous treatment was performed with angioplasty, and the patient was discharged with dual antiplatelet therapy. It is important to be aware of this type of clinical presentation, since the diagnosis can be confused with other diseases that produce similar signs and symptoms. It would be ideal to have a high clinical suspicion in order to avoid delays in the management.

Tumores de Sistema Nervioso Central en Pediatría: Presente y Futuro del Abordaje Diagnóstico / Central nervous system tumors in pediatrics: present and future of diagnostic approach

Resumen Los tumores de sistema nervioso central son neoplasias con alta frecuencia durante la infancia, con una incidencia que alcanza los 31,2 casos por millón de niños, siendo el más común de estos el astrocitoma. Estos son producto de una combinación de factores genéticos y epigenéticos que determinan no solo su aparición, sino también, el comportamiento y pronóstico del mismo. La presentación clínica de estas neoplasias va desde súbita y clara, hasta larvada y ambigua, en ocasiones, dificultando realizar un adecuado abordaje y un diagnóstico precoz, por lo cual se han destinado muchos recursos para la evaluación y estandarización de nuevos métodos diagnósticos como lo son los marcadores tumorales. Objetivo: Ofrecer una revisión clara acerca de los tumores de sistema nervioso central y nuevas perspectivas en el diagnóstico. Se llevó a cabo una búsqueda de literatura en bases de datos empleando los descriptores sistema nervioso central, neoplasias, astrocitoma, se incluyeron 25 artículos en español y en inglés. Los tumores de sistema nervioso central cobran una relevancia notable en la población pediátrica, su comportamiento y pronóstico hacen necesaria la búsqueda de nuevas ayudas que permitan un diagnóstico precoz.

Abstract Central nervous system tumors are neoplasms with high frequency during childhood, with an incidence of 31,2 cases per million children, the most common being astrocytoma. These are the product of a combination of genetic and epigenetic factors that determine not only their appearance, but also their behavior and prognosis. The clinical presentation of these neoplasms ranges from the sudden and clear, to the gradual and ambiguous, which represents a barrier to carrying out an adequate approach and an early diagnosis. That is the reason to destinate many resources for evaluation and standarization of new diagnostic methods like tumor markers. Objective: To provide a clear review of central nervous system tumors and new perspectives in diagnosis. A search of the literature was carried out in databases using the descriptors central nervous system, neoplasms, astrocytoma. 25 articles were included in spanish and english. Central nervous system tumors have a significant relevance in the pediatric population. Their behavior and prognosis necessitate searching for new aids for an early diagnosis.

Individual prognosis of abutment teeth / Pronóstico individual de dientes pilares

ABSTRACT. Establishing accurate prognosis after correct diagnosis is the most important point of reference in our clinical practice. The need to correctly predicting teeth permanence in the mouth has prompted each dental discipline to find ways to establish it. Since prosthodontics, or oral rehabilitation, incorporates the procedures of other disciplines and usually is the one that finishes most treatments, it should review all the factors affecting the prognosis of comprehensive treatment. Below is a review of dental and periodontal considerations for prognosis in prosthodontics, as well as the implications of treatments performed on abutment teeth in order to retain them, and the strategic weight of each abutment.

RESUMEN. El establecimiento de un acertado pronóstico, luego de un correcto diagnóstico, es el más importante orientador de nuestra conducta en la práctica clínica. La necesidad de predecir, de la forma más audaz posible, la supervivencia de los dientes en boca, ha hecho que cada disciplina odontológica se acerque de una manera u otra a su establecimiento. La prostodoncia o rehabilitación oral, en la medida en que integra los procedimientos de las otras disciplinas y culmina la mayoría de tratamientos, debe revisar todos los factores que modifican el pronóstico del tratamiento integral. A continuación se revisan las consideraciones dentales y periodontales del pronóstico en prostodoncia, así como las implicaciones de los tratamientos que se le hagan a un diente pilar para ser conservado y el peso estratégico de cada pilar.