Case Report: Interferon- γ Rescues Monocytic Human Leukocyte Antigen Receptor (mHLA-DR) Function in a COVID-19 Patient With ARDS and Superinfection With Multiple MDR 4MRGN Bacterial Strains.

Background: CD14+ monocytes present antigens to adaptive immune cells via monocytic human leukocyte antigen receptor (mHLA-DR), which is described as an immunological synapse. Reduced levels of mHLA-DR can display an acquired immune defect, which is often found in sepsis and predisposes for secondary infections and fatal outcomes. Monocytic HLA-DR expression is reliably induced by interferon- γ (IFNγ) therapy. Case Report: We report a case of multidrug-resistant superinfected COVID-19 acute respiratory distress syndrome (ARDS) on extracorporeal membrane oxygenation (ECMO) support. The resistance profiles of the detected Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii and Citrobacter freundii isolates were equipped with resistance to all four antibiotic classes including carbapenems (4MRGN) and Cefiderocol in the case of K. pneumoniae. A causal therapeutic antibiotic strategy was not available. Therefore, we measured the immune status of the patient aiming to identify a potential acquired immune deficiency. Monocyte HLA-DR expression identified by FACS analysis revealed an expression level of 34% positive monocytes and suggested severe immunosuppression. We indicated IFNγ therapy, which resulted in a rapid increase in mHLA-DR expression (96%), rapid resolution of invasive bloodstream infection, and discharge from the hospital on day 70. Discussion: Superinfection is a dangerous complication of COVID-19 pneumonia, and sepsis-induced immunosuppression is a risk factor for it. Immunosuppression is expressed by a disturbed antigen presentation of monocytes to cells of the adaptive immune system. The case presented here is remarkable as no validated antibiotic regimen existed against the detected bacterial pathogens causing bloodstream infection and severe pneumonia in a patient suffering from COVID-19 ARDS. Possible restoration of the patient's own immunity by IFNγ was a plausible option to boost the patient's immune system, eliminate the identified 4MRGNs, and allow for lung recovery. This led to the conclusion that immune status monitoring is useful in complicated COVID-19-ARDS and that concomitant IFNγ therapy may support antibiotic strategies. Conclusion: After a compromised immune system has been detected by suppressed mHLA-DR levels, the immune system can be safely reactivated by IFNγ.

Development of Complement Factor H-Based Immunotherapeutic Molecules in Tobacco Plants Against Multidrug-Resistant Neisseria gonorrhoeae.

Novel therapeutics against the global threat of multidrug-resistant Neisseria gonorrhoeae are urgently needed. Gonococci possess several mechanisms to evade killing by human complement, including binding of factor H (FH), a key inhibitor of the alternative pathway. FH comprises 20 short consensus repeat (SCR) domains organized in a head-to-tail manner as a single chain. N. gonorrhoeae binds two regions in FH; domains 6 and 7 and domains 18 through 20. We designed a novel anti-infective immunotherapeutic molecule that fuses domains 18-20 of FH containing a D-to-G mutation in domain 19 at position 1119 (called FH*) with human IgG1 Fc. FH*/Fc retained binding to gonococci but did not lyse human erythrocytes. Expression of FH*/Fc in tobacco plants was undertaken as an alternative, economical production platform. FH*/Fc was expressed in high yields in tobacco plants (300-600 mg/kg biomass). The activities of plant- and CHO-cell produced FH*/Fc against gonococci were similar in vitro and in the mouse vaginal colonization model of gonorrhea. The addition of flexible linkers [e.g., (GGGGS)2 or (GGGGS)3] between FH* and Fc improved the bactericidal efficacy of FH*/Fc 2.7-fold. The linkers also improved PMN-mediated opsonophagocytosis about 11-fold. FH*/Fc with linker also effectively reduced the duration and burden of colonization of two gonococcal strains tested in mice. FH*/Fc lost efficacy: i) in C6-/- mice (no terminal complement) and ii) when Fc was mutated to abrogate complement activation, suggesting that an intact complement was necessary for FH*/Fc function in vivo. In summary, plant-produced FH*/Fc represent promising prophylactic or adjunctive immunotherapeutics against multidrug-resistant gonococci.

Development of A4 antibody for detection of neuraminidase I223R/H275Y-associated antiviral multidrug-resistant influenza virus.

The emergence and spread of antiviral drug-resistant viruses have been a worldwide challenge and a great concern for patient care. We report A4 antibody specifically recognizing and binding to the mutant I223R/H275Y neuraminidase and prove the applicability of A4 antibody for direct detection of antiviral multidrug-resistant viruses in various sensing platforms, including naked-eye detection, surface-enhanced Raman scattering-based immunoassay, and lateral flow system. The development of the A4 antibody enables fast, simple, and reliable point-of-care assays of antiviral multidrug-resistant influenza viruses. In addition to current influenza virus infection testing methods that do not provide information on the antiviral drug-resistance of the virus, diagnostic tests for antiviral multidrug-resistant viruses will improve clinical judgment in the treatment of influenza virus infections, avoid the unnecessary prescription of ineffective drugs, and improve current therapies.

Tumor microenvironment and epithelial mesenchymal transition as targets to overcome tumor multidrug resistance.

It is well established that multifactorial drug resistance hinders successful cancer treatment. Tumor cell interactions with the tumor microenvironment (TME) are crucial in epithelial-mesenchymal transition (EMT) and multidrug resistance (MDR). TME-induced factors secreted by cancer cells and cancer-associated fibroblasts (CAFs) create an inflammatory microenvironment by recruiting immune cells. CD11b+/Gr-1+ myeloid-derived suppressor cells (MDSCs) and inflammatory tumor associated macrophages (TAMs) are main immune cell types which further enhance chronic inflammation. Chronic inflammation nurtures tumor-initiating/cancer stem-like cells (CSCs), induces both EMT and MDR leading to tumor relapses. Pro-thrombotic microenvironment created by inflammatory cytokines and chemokines from TAMs, MDSCs and CAFs is also involved in EMT and MDR. MDSCs are the most common mediators of immunosuppression and are also involved in resistance to targeted therapies, e.g. BRAF inhibitors and oncolytic viruses-based therapies. Expansion of both cancer and stroma cells causes hypoxia by hypoxia-inducible transcription factors (e.g. HIF-1α) resulting in drug resistance. TME factors induce the expression of transcriptional EMT factors, MDR and metabolic adaptation of cancer cells. Promoters of several ATP-binding cassette (ABC) transporter genes contain binding sites for canonical EMT transcription factors, e.g. ZEB, TWIST and SNAIL. Changes in glycolysis, oxidative phosphorylation and autophagy during EMT also promote MDR. Conclusively, EMT signaling simultaneously increases MDR. Owing to the multifactorial nature of MDR, targeting one mechanism seems to be non-sufficient to overcome resistance. Targeting inflammatory processes by immune modulatory compounds such as mTOR inhibitors, demethylating agents, low-dosed histone deacetylase inhibitors may decrease MDR. Targeting EMT and metabolic adaptation by small molecular inhibitors might also reverse MDR. In this review, we summarize evidence for TME components as causative factors of EMT and anticancer drug resistance.

Caracterización de las úlceras por presión en adultos con gérmenes multirresistentes / Characterization of pressure lesions in adults with multidrug resistant germs / Caracterização das lesões por pressão em adultos portadores de germes multirresistentes

OBJETIVO: Describir las características de las lesiones por presión en pacientes adultos con gérmenes multirresistentes. MÉTODO: Este es un estudio transversal realizado con pacientes hospitalizados en la unidad de pacientes hospitalizados por gérmenes resistentes a múltiples fármacos de un hospital público en el sur de Brasil. Se seleccionaron pacientes con lesión por presión del estadio II. Los datos se recopilaron en 2017 de una muestra de 110 lesiones en 36 pacientes utilizando la Herramienta de evaluación de heridas Bates-Jensen (BWAT). Los datos se analizaron mediante estadísticas descriptivas y analíticas. RESULTADOS: La edad promedio de los individuos fue de 45.4 (± 21.3) años y el 89.1% había sido hospitalizado con una lesión por presión, que ocurrió en el hogar o en otras instituciones de salud. El valor medio de BWAT fue de 35.5 ± 8.9 puntos y hubo una correlación positiva débil (r = 0.228 p = 0.017) con el tamaño de la lesión, correlación positiva moderada con el estadio de la lesión (r = 540 p <0.001). y con el resultado de la escala de Braden (r = 0.44 p = 0.651). CONCLUSIÓN: Los resultados muestran la enfermedad de pacientes jóvenes. Los pacientes con gérmenes multirresistentes presentaron lesiones por presión con una mayor participación de las estructuras, lo que sugiere la necesidad de apoyo en el hogar

OBJECTIVE: To describe the characteristics of the pressure lesions in adult patients with multiresistant germs. METHOD: This is a cross-sectional study conducted with patients admitted to the inpatient unit for multidrug resistant germs in a public hospital in Brazil. Patients with pressure lesions from stage II were selected. Data collection took place in 2017, in a sample of 110 injuries, in patients, through the Pressure Ulcer State Assessment Instrument (Bates-Jensen Wound Assessment Tool - BWAT). RESULTS: The average age of the individuals was 45.4 (± 21.3) years old and 89.1% had already suffered pressure lesions, such as those that occurred at home or in other health institutions. The mean BWAT value was 35.5 ± 8.9 points and there was a weak positive correlation (r = 0.228 p = 0.017) with lesion size, moderate positive correlation with lesion stage (r = 540 p <0.001), and with the result of the Braden scale (r = 0.44 p = 0.651). CONCLUSION: The results indicated the illness of young patients. Patients with multiresistant germs suffered pressure lesions with greater involvement of structures, which suggests the need for home support

OBJETIVO: Descrever as características das lesões por pressão em pacientes adultos portadores de germes multirresistentes. MÉTODO: Trata-se de um estudo transversal, realizado com pacientes hospitalizados na unidade de internação para portadores de germes multirresistentes de um hospital público do sul do Brasil. Foram selecionados pacientes portadores de lesão por pressão a partir de estágio II. A coleta de dados ocorreu em 2017, em uma amostra de 110 lesões, através do Instrumento de Avaliação do estado da Úlcera por Pressão (Bates-Jensen Wound Assessment Tool - BWAT. Os dados foram analisados através de estatística descritiva e analítica. RESULTADOS: A idade média dos indivíduos foi 45,4 (± 21,3) anos e 89,1% já internaram com lesão por pressão, as quais ocorreram no domicílio ou em outras instituições de saúde. O valor médio da BWAT foi de 35,5 ± 8,9 pontos e houve correlação positiva fraca (r=0,228 p = 0,017) com o tamanho da lesão, correlação positiva moderada com o estágio da lesão (r= 540 p < 0,001) e com o resultado da escala de Braden (r= 0,44 p = 0,651). CONCLUSÃO: Os resultados apontam o adoecimento de pacientes jovens. Pacientes portadores de germes multirresistentes apresentaram lesões por pressão com maior acometimento de estruturas, o que sugere necessidade de aporte domiciliar

Colonización por ESKAPES y características clínicas de pacientes en estado crítico / Colonization by ESKAPES and clinical characteristics of critically ill patients / Colonização por ESKAPES e características clínicas de pacientes críticos

OBJETIVO: Identificar la colonización por ESKAPES y las características clínicas de los pacientes hospitalizados en una Unidad de Cuidados Intensivos para Adultos de un hospital mixto en Paraná. MÉTODO: Investigación de campo, descriptiva, documental y experimental con enfoque cuantitativo, desarollada en una Unidad de Cuidados Intensivos adultos de un hospital mixto en el suroeste de Paraná, Brasil. La población del estudio consistió en pacientes con ingreso de 48 horas en la Unidad de Cuidados Intensivos, de abril a agosto de 2018 y de abril a agosto de 2019. La muestra totalizó 102 individuos. Para la recopilación de datos clínicos, se utilizó un Checklist y para el análisis microbiológico se recogieron muestras de las cavidades nasales y orales y la secreción traqueal. El análisis de los datos clínicos se produjo a través del software Statistical Package for the Social Sciences. Se realizaron pruebas de frecuencia y chi-cuadrado, teniendo en cuenta la p < 0,05 significativa. RESULTADOS: Se evaluaron un total de 102 pacientes ingresados en la Unidad de Cuidados Intensivos durante el período estudiado. De ellos, 57 (55,8%) fueron colonizados por microorganismos patógenos. En cuanto a la colonización por microorganismos, predominan Staphylococcus aureus (61,4%), seguido de Klebsiella pneumoniae (40,4%), Pseudomonas aeruginosa (26,3%) y Staphylococcus epidermidis (21,1%). Cabe destacar que Klebsiella pneumoniae y Staphylococcus aureus estuvieron presentes en las tres regiones evaluadas. CONCLUSIÓN: El estudio identificó la presencia de colonización en pacientes en estado crítico estudiados, siendo esta colonización, en su mayoría, por bacterias resistentes pertenecientes al grupo ESKAPE

OBJECTIVE: To identify colonization by ESKAPES and clinical characteristics of patients admitted in Adult Intensive Care Unit of a mixed hospital in Paraná. METHOD: Field research, descriptive, documentary and experimental quantitative approach, developed in adult Intensive Care Unit of a mixed hospital in Southwest Paraná, Brazil. The study population consisted of patients with admission from 48 hours in the Intensive Care Unit, from April to August 2018 and April to August 2019. The sample has 102 individuals. For the collection of clinical data, a checklist was used and for microbiological analysis the sample was collected from nasal and oral cavities and tracheal secretion. The analysis of clinical data occurred through the Statistical Package for the Social Sciences software. Descriptive frequency and chi-square test, considering significant p <0,05. RESULTS: A total of 102 patients admitted to the Intensive Care Unit during the period studied were evaluated. On these ones, 57 (55,8%) were colonized by pathogenic microorganisms. Regarding the colonization of microorganisms, there was predominance of Staphylococcus aureus (61,4%), followed by Klebsiella pneumoniae (40,4%), Pseudomonas aeruginosa (26,3%) and Staphylococcus epidermidis (21,1%). It is noteworthy that Klebsiella pneumoniae and Staphylococcus aureus were present in the three regions evaluated. CONCLUSION: The study identified the presence of colonization in critically ill patients studied, being this colonization, mostly, resistant bacteria belonging to the ESKAPE group

OBJETIVO: Identificar a colonização por ESKAPES e características clínicas de pacientes internados em uma Unidade de Terapia Intensiva Adulto de um hospital misto do Paraná. MÉTODO: Pesquisa de campo, descritiva, documental e experimental com abordagem quantitativa, desenvolvida em uma Unidade de Terapia Intensiva adulto de um hospital misto do Sudoeste do Paraná, Brasil. A população do estudo constituiu-se pelos pacientes com admissão a partir de 48 horas na Unidade de Terapia Intensiva, no período de abril a agosto de 2018 e de abril a agosto de 2019. A amostra totalizou 102 indivíduos. Para a coleta de dados clínicos foi utilizado um Checklist e para a análise microbiológica foram coletadas amostras das cavidades nasal e oral e secreção traqueal. A análise dos dados ocorreu por meio do software Statistical Package for the Social Sciences. Realizou-se frequência descritiva e teste de qui-quadrado, considerando significativo p <0,05. RESULTADOS: Foram avaliados 102 pacientes admitidos na Unidade de Terapia Intensiva durante o período pesquisado. Destes, 57 (55,8%) estavam colonizados por microrganismos patogênicos. Em relação à colonização de microrganismos, houve predominância de Staphylococcus aureus (61,4%), seguido por Klebsiella pneumoniae (40,4%), Pseudomonas aeruginosa (26,3%) e Staphylococcus epidermidis (21,1%). Vale ressaltar que, Klebsiella pneumoniae e Staphylococcus aureus estiveram presentes nas três regiões avaliadas. CONCLUSÃO: O estudo identificou a presença de colonização nos pacientes criticamente enfermos pesquisados, sendo essa colonização, em sua maioria, por bactérias resistentes pertencentes ao grupo ESKAPE

Novel Antimicrobial Peptides Formulated in Chitosan Matrices are Effective Against Biofilms of Multidrug-Resistant Wound Pathogens.

INTRODUCTION: Infection frequently complicates the treatment of combat-related wounds, impairs healing, and leads to worse outcomes. To better manage wound infections, antimicrobial therapies that are effective against biofilm and designed for direct wound application are needed. The primary objective of this work was to evaluate a chitosan matrix for delivery of two engineered antimicrobial peptides, (ASP)-1 and ASP-2, to treat biofilm-associated bacteria. A secondary objective was to determine whether replacing the levorotatory (L) form amino acids in ASP-2 with dextrorotatory (D) form amino acids would impact peptide activity. MATERIALS AND METHODS: Chitosan gels loaded with antimicrobial peptides were evaluated for peptide release over 7 days and tested for efficacy against biofilms grown both in vitro on polymer mesh and ex vivo on porcine skin. RESULTS: When delivered via chitosan, 70% to 80% of peptides were released over 7 days. Gels eradicated biofilms of gram-positive and gram-negative, drug-resistant bacteria in vitro and ex vivo. Under the conditions tested, no meaningful differences in peptide activity between the L and D forms of ASP-2 were detected. CONCLUSIONS: Chitosan serves as an effective delivery platform for ASP-1 and ASP-2 to treat biofilm-embedded bacteria and warrants further development as a topical treatment.

Assessment and Management of Infection in Alcoholic Hepatitis.

Severe alcoholic hepatitis (SAH) is a condition characterized by jaundice and liver failure that develops after heavy and prolonged alcohol consumption. Infection frequently complicates the natural history of the disease and is independently associated with mortality. Objective recognition and recording of infection are therefore essential in the evaluation of therapeutic interventions and for antibiotic stewardship. This review will evaluate infections that complicate SAH at admission and beyond. Factors that associate with the development of infection will be identified and clinical and laboratory techniques available to identify infection will be discussed. Common pathogens and frequently used antibiotics will be reviewed and recommendations will be made for the management of infection for SAH patients. New techniques to assess infection earlier and more precisely may improve diagnosis and treatment of this important driver of mortality in SAH.

Insight on Multidrug Resistance and Nanomedicine Approaches to Overcome MDR.

Multidrug resistance (MDR) remains a major obstacle to ensure effective chemotherapy in cancer patients. Several factors could be associated with cancer cells' drug resistance such as overexpression of P-glycoprotein (P-gp), cancer stem cells (CSCs), defect in apoptosis, mutation and alteration in DNA repair pathways, angiogenesis, autophagy, and modulation in metabolic enzymes. Until now, drug efflux by ABC transporters has been a univocal and well-established mechanism of chemotherapeutic associated drug resistance. To explore the mechanics involved in ABC transporter associated drug resistance, many crucial studies have been conducted from identification of drug binding sites to elucidation of their structure. Due to our continuous battle with drug resistance, several strategies have been employed to combat MDR, including P-gp modulators, siRNAs, antibodies, as well as peptides. Furthermore, various nanoparticle and different effective combination nanomedicine strategies also suggest some exciting results. Thus, to improve nanomedicine approaches to overcome MDR, in this evolutionary review, we have focused on fundamentals of possible strategies as well as the latest accomplishments to reverse MDR.

Task-Specific Design of Immune-Augmented Nanoplatform to Enable High-Efficiency Tumor Immunotherapy.

Potentiating systemic immunity against breast cancer is in the most urgent demand as breast cancer is less sensitive to immune checkpoint blockade. Although phototherapy and some chemotherapy can trigger immunogenic cell death (ICD) for T cell-mediated antitumor immune response, their immunotherapy efficacy is severely restricted by insufficient phototherapeutic capability and severe multidrug resistance (MDR). Inspired by both the hypersensitivity to phototherapy and the key role of MDR for mitochondria, a rationally engineered immunity amplifier via mitochondria-targeted photochemotherapeutic nanoparticles was, for the first time, achieved to fight against low-immunogenic breast cancer without additional immune agents. The newly synthesized task-specific mitochondria-targeted IR780 derivative (T780) was integrated with chemotherapeutic doxorubicin (DOX) to form multifunctional nanoparticles via an assembling strategy along with bovine serum albumin (BSA) as a biomimetic corona (BSA@T780/DOX NPs). The in situ enhancement in both phototherapy and MDR reversal by targeting mitochondria with BSA@T780/DOX NPs boosted highly efficient ICD toward excellent antitumor immune response. The newly developed strategy not only eradicated the primary tumor but also eliminated the bilateral tumors efficiently, as well as preventing metastasis and postsurgical recurrence, demonstrating great interest for fighting against low-immunogenic breast cancer.

The NDV-3A vaccine protects mice from multidrug resistant Candida auris infection.

Candida auris is an emerging, multi-drug resistant, health care-associated fungal pathogen. Its predominant prevalence in hospitals and nursing homes indicates its ability to adhere to and colonize the skin, or persist in an environment outside the host-a trait unique from other Candida species. Besides being associated globally with life-threatening disseminated infections, C. auris also poses significant clinical challenges due to its ability to adhere to polymeric surfaces and form highly drug-resistant biofilms. Here, we performed bioinformatic studies to identify the presence of adhesin proteins in C. auris, with sequence as well as 3-D structural homologies to the major adhesin/invasin of C. albicans, Als3. Anti-Als3p antibodies generated by vaccinating mice with NDV-3A (a vaccine based on the N-terminus of Als3 protein formulated with alum) recognized C. auris in vitro, blocked its ability to form biofilms and enhanced macrophage-mediated killing of the fungus. Furthermore, NDV-3A vaccination induced significant levels of C. auris cross-reactive humoral and cellular immune responses, and protected immunosuppressed mice from lethal C. auris disseminated infection, compared to the control alum-vaccinated mice. The mechanism of protection is attributed to anti-Als3p antibodies and CD4+ T helper cells activating tissue macrophages. Finally, NDV-3A potentiated the protective efficacy of the antifungal drug micafungin, against C. auris candidemia. Identification of Als3-like adhesins in C. auris makes it a target for immunotherapeutic strategies using NDV-3A, a vaccine with known efficacy against other Candida species and safety as well as efficacy in clinical trials. Considering that C. auris can be resistant to almost all classes of antifungal drugs, such an approach has profound clinical relevance.

Chloroquine and mefloquine resistance profiles are not related to the circumsporozoite protein (CSP) VK210 subtypes in field isolates of Plasmodium vivax from Manaus, Brazilian Amazon

BACKGROUND The central repetitive region (CRR) of the Plasmodium vivax circumsporozoite surface protein (CSP) is composed of a repetitive sequence that is characterised by three variants: VK210, VK247 and P. vivax-like. The most important challenge in the treatment of P. vivax infection is the possibility of differential response based on the parasite genotype. OBJECTIVES To characterise the CSP variants in P. vivax isolates from individuals residing in a malaria-endemic region in Brazil and to profile these variants based on sensitivity to chloroquine and mefloquine. METHODS The CSP variants were determined by sequencing and the sensitivity of the P. vivax isolates to chloroquine and mefloquine was determined by Deli-test. FINDINGS Although five different allele sizes were amplified, the sequencing results showed that all of the isolates belonged to the VK210 variant. However, we observed substantial genetic diversity in the CRR, resulting in the identification of 10 different VK210 subtypes. The frequency of isolates that were resistant to chloroquine and mefloquine was 11.8 and 23.8%, respectively. However, we did not observe any difference in the frequency of the resistant isolates belonging to the VK210 subtypes. MAIN CONCLUSION The VK210 variant is the most frequently observed in the studied region and there is significant genetic variability in the CRR of the P. vivax CSP. Moreover, the antimalarial drug sensitivity profiles of the isolates does not seem to be related to the VK210 subtypes.

Stabilization of HDAC1 via TCL1-pAKT-CHFR axis is a key element for NANOG-mediated multi-resistance and stem-like phenotype in immune-edited tumor cells.

Cancer immunoediting enriches NANOG expression in tumor cells, resulting in multi-drug resistance and stem-like phenotypes. We previously demonstrated that these NANOG-associated phenotypes are promoted through HDAC1 transcriptional upregulation. In this study, we identified that NANOG also contributes to the stabilization of HDAC1 protein through the AKT signaling pathway. NANOG-AKT axis leads to phosphor-dependent inactivation of CHFR, an E3 ligase for HDAC1 protein, and thereby inhibiting the ubiquitin-mediated degradation of HDAC1. Furthermore, AKT inhibition disrupts HDAC1 WT-mediated phenotypes but had no effect on the phenotypes mediated by HDAC1 FM, a mutant that is unable to interact with CHFR. Critically, we applied a catalytic dead mutant, HDAC1-H141A, to uncover that HDAC1 confers immune-resistance, drug-resistance and stem-like phenotype in tumor cells through its catalytic activity. Collectively, our results establish a firm molecular link in immune-edited tumor cells among NANOG, AKT, CHFR, and HDAC1, identifying HDAC1 as a molecular target in controlling NANOGHIGH immune-refractory cancer.

Microbiota Replacement Therapies: Innovation in Gastrointestinal Care.

There has been an increasing interest in the association between human disease and altered gut microbiota, and therapeutics to modulate microbiota to treat disease. Healthy human gastrointestinal microbiota is highly diverse and rich, and harbors between 500 and 2,000 species. Diseases associated with dysbiotic microbiota include antibiotic-associated diarrhea, Clostridium difficile infection, multidrug-resistant organisms, inflammatory bowel disease, obesity, metabolic syndrome, diabetes mellitus, neuropsychiatric diseases, and systemic autoimmune diseases. Microbiota replacement therapies have shown immense promise in treatment of recurrent C. difficile infection and are being studied for other indications. Microbiota replacement therapies for indications other than C. difficile infection should be performed only in research settings. There is an immense need for standardized microbiota replacement therapies for C. difficile infection. Studies are needed to elucidate long-term safety and adverse events from these therapies.

Natural killer cell-intrinsic type I IFN signaling controls Klebsiella pneumoniae growth during lung infection.

Klebsiella pneumoniae is a significant cause of nosocomial pneumonia and an alarming pathogen owing to the recent isolation of multidrug resistant strains. Understanding of immune responses orchestrating K. pneumoniae clearance by the host is of utmost importance. Here we show that type I interferon (IFN) signaling protects against lung infection with K. pneumoniae by launching bacterial growth-controlling interactions between alveolar macrophages and natural killer (NK) cells. Type I IFNs are important but disparate and incompletely understood regulators of defense against bacterial infections. Type I IFN receptor 1 (Ifnar1)-deficient mice infected with K. pneumoniae failed to activate NK cell-derived IFN-γ production. IFN-γ was required for bactericidal action and the production of the NK cell response-amplifying IL-12 and CXCL10 by alveolar macrophages. Bacterial clearance and NK cell IFN-γ were rescued in Ifnar1-deficient hosts by Ifnar1-proficient NK cells. Consistently, type I IFN signaling in myeloid cells including alveolar macrophages, monocytes and neutrophils was dispensable for host defense and IFN-γ activation. The failure of Ifnar1-deficient hosts to initiate a defense-promoting crosstalk between alveolar macrophages and NK cell was circumvented by administration of exogenous IFN-γ which restored endogenous IFN-γ production and restricted bacterial growth. These data identify NK cell-intrinsic type I IFN signaling as essential driver of K. pneumoniae clearance, and reveal specific targets for future therapeutic exploitations.

Immunotherapy for triple-negative breast cancer: Existing challenges and exciting prospects.

Patients with breast tumors that do not express the estrogen receptor, the progesterone receptor, nor Her-2/neu are hence termed "triple negatives", and generally have a poor prognosis, with high rates of systemic recurrence and refractoriness to conventional therapy regardless of the choice of adjuvant treatment. Thus, more effective therapeutic options are sorely needed for triple-negative breast cancer (TNBC), which occurs in approximately 20% of diagnosed breast cancers. In recent years, exploiting intrinsic mechanisms of the host immune system to eradicate cancer cells has achieved impressive success, and the advances in immunotherapy have yielded potential new therapeutic strategies for the treatment of this devastating subtype of breast cancer. It is anticipated that the responses initiated by immunotherapeutic interventions will explicitly target and annihilate tumor cells, while at the same time spare normal cells. Various immunotherapeutic approaches have been already developed and tested, which include the blockade of immune checkpoints using neutralizing or blocking antibodies, induction of cytotoxic T lymphocytes (CTLs), adoptive cell transfer-based therapy, and modulation of the tumor microenvironment to enhance the activity of CTLs. One of the most important areas of breast cancer research today is understanding the immune features and profiles of TNBC and devising novel immune-modulatory strategies to tackling TNBC, a subtype of breast cancer notorious for its poor prognosis and its imperviousness to conventional treatments. On the optimal side, one can anticipate that novel, effective, and personalized immunotherapy for TNBC will soon achieve more success and impact clinical treatment of this disease which afflicts approximately 20% of patients with breast cancer. In the present review, we highlight the current progress and encouraging developments in cancer immunotherapy, with a goal to discuss the challenges and to provide future perspectives on how to exploit a variety of new immunotherapeutic approaches including checkpoint inhibitors and neoadjuvant immunotherapy for the treatment of patients with TNBC.

HDAC1 Upregulation by NANOG Promotes Multidrug Resistance and a Stem-like Phenotype in Immune Edited Tumor Cells.

Cancer immunoediting drives the adaptation of tumor cells to host immune surveillance. Immunoediting driven by antigen (Ag)-specific T cells enriches NANOG expression in tumor cells, resulting in a stem-like phenotype and immune resistance. Here, we identify HDAC1 as a key mediator of the NANOG-associated phenotype. NANOG upregulated HDAC1 through promoter occupancy, thereby decreasing histone H3 acetylation on K14 and K27. NANOG-dependent, HDAC1-driven epigenetic silencing of cell-cycle inhibitors CDKN2D and CDKN1B induced stem-like features. Silencing of TRIM17 and NOXA induced immune and drug resistance in tumor cells by increasing antiapoptotic MCL1. Importantly, HDAC inhibition synergized with Ag-specific adoptive T-cell therapy to control immune refractory cancers. Our results reveal that NANOG influences the epigenetic state of tumor cells via HDAC1, and they encourage a rational application of epigenetic modulators and immunotherapy in treatment of NANOG+ refractory cancer types. Cancer Res; 77(18); 5039-53. ©2017 AACR.

Caracterización de mecanismos de resistencia a carbapenémicos en aislados clínicos de Pseudomonas aeruginosa en un hospital español / Characterisation of carbapenem-resistance mechanisms in clinical Pseudomonas aeruginosa isolates recovered in a Spanish hospital

INTRODUCCIÓN: Los carbapenémicos son los antibióticos betalactámicos con mayor espectro de actividad en el tratamiento de infecciones por Pseudomonas aeruginosa. El objetivo de este trabajo fue caracterizar molecularmente una colección de aislados de P. aeruginosa resistentes a carbapenémicos (PARC). MÉTODOS: Se recogieron 85 aislados PARC de 60 pacientes en el Hospital San Pedro, Logroño (período 2008-2011). La relación clonal se determinó por electroforesis en gel de campo pulsado (PFGE), la sensibilidad a 15 antipseudomónicos por método de difusión en disco y las alteraciones en oprD, la caracterización de integrones y la tipificación molecular (MLST) por PCR y secuenciación. RESULTADOS: Las 85 PARC se clasificaron en 35 perfiles diferentes de PFGE. Se seleccionaron 61 cepas de los 60 pacientes y se observó que eran multirresistentes, aunque ninguna mostró fenotipo carbapenemasa. Se detectó un gran polimorfismo de OprD, destacando que el 59% de las cepas presentaban un codón de finalización prematuro. ISPa1328 e ISPsp4 truncaban el gen oprD en 2 cepas (GenBank KF517097 y KF517098). El 67% de las cepas presentó integrones de clase 1 con genes codificantes de enzimas modificantes de aminoglucósidos, 2 de las cuales portaban un nuevo integrón: aac(3)-Ia+aadA1h (nombrado In272, GenBank GQ144317). Se detectaron 4 secuencias tipo (ST) (número de cepas): ST175 (35), ST308 (3), ST235 (2) y ST639 (1). CONCLUSIÓN: La multirresistencia, el alto polimorfismo de oprD, el alto porcentaje de integrones, la moderada relación clonal de las cepas y la elevada diseminación epidémica de clones de alto riesgo son aspectos de gran preocupación clínica para erradicar la diseminación de PARC

INTRODUCTION: Carbapenems are the beta-lactam antibiotics with the best spectrum of activity in the treatment of Pseudomonas aeruginosa infections. The objective of this study was to molecularly characterise a collection of carbapenem-resistant P. aeruginosa isolates (PARC). METHODS: A total of 85 PARC isolates were recovered from 60 patients in the Hospital San Pedro, Logroño (period 2008-2011). Clonal relationship was determined using pulsed-field gel electrophoresis (PFGE), susceptibility testing to 15 anti-pseudomonal agents was performed using the disk diffusion method, and alterations in oprD, characterisation of integrons and molecular typing (MLST) using PCR and sequencing. RESULTS: The 85 PARC were classified into 35 different PFGE profiles. Of the 61 selected strains from 60 patients all of them were multiresistant, although none of them showed a carbapenemase phenotype. High polymorphism was detected in OprD, emphasising that 59% of the strains had a premature stop codon. ISPa1328 and ISPsp4 insertion sequences truncated oprD gene into 2 strains (GenBank KF517097 and KF517098). Two-thirds (67%) of the strains showed class 1 integrons with genes encoding aminoglycoside modifying enzymes, and 2 of them carried a new integron: aac(3)-Ia+aadA1h, named In272, GenBank GQ144317. Four sequence types were detected (Strain Nos.): ST175 (35), ST308 (3), ST235 (2), and ST639 (1). CONCLUSION: Multidrug resistance, high polymorphism in oprD, a high percentage of integrons, moderate clonal relationship of strains, and the high epidemic dissemination of high-risk clones are clinical aspects of great concern in order to eradicate the spread of PARC

Multidrug Resistance in Non-PCV13 Serotypes of Streptococcus pneumoniae in Northern Japan, 2014.

Since the implementation of routine PCV13 immunization in Japan, nonvaccine serotypes (NVTs) have been increasing among clinical isolates of Streptococcus pneumoniae. In this study, susceptibility to 18 antibiotics was tested for all the 231 isolates with NVTs, which were collected from children <16 years of age in northern Japan in 2014 (July-November). High resistance rates were observed for macrolides (>90.9%), tetracycline (91.3%), and clindamycin (75.3%), while penicillin (PEN) nonsusceptibility (PNSP; MIC ≥0.12 µg/ml) was detected in 42.9% of the pneumococci [39.4%; PEN-intermediate S. pneumoniae (PISP), 3.5%; PEN-resistant S. pneumoniae (PRSP)]. All serotype 15A isolates were PRSP (MIC, ≥2 µg/ml) or PISP, and PNSP was prevalent in also serotypes 23A (96.9%), 6C (41%), and 35B (33.3%). Overall, 42.0% of the isolates showed multidrug resistance (MDR). Sequence types (STs) determined for 20 PNSP isolates with NVTs were ST63 (15A), STs 242 or 5832 (6C), STs 338 or 5242 (23A), and ST558 (35B). All the PNSP isolates possessed tet(M), and erm(B) or mefA(A/E), and 70% of them were gPRSP having three altered genes pbp1a, pbp2x, and pbp2b. Among alterations in transpeptidase-coding region of penicillin-binding proteins (PBPs), two substitutions of T371S in the STMK motif and TSQF574-577NTGY in PBP1a were common to all PRSP isolates. The present study showed the spread of PNSP in NVTs 15A, 23A, 6C, and 35B, and the emergence of the MDR international clone Sweden15A-ST63 in northern Japan.

Brote de Enterobacter cloacae complex multirresistente productor de CTX-M-9 en una unidad de cuidados intensivos / Outbreak of multidrug-resistant CTX-M-9-producing Enterobacter cloacae complex in an intensive care unit

INTRODUCCIÓN: Descripción clínica y epidemiológica de un brote en una unidad de cuidados intensivos (UCI) causado por Enterobacter cloacae complex multirresistente productor de una beta-lactamasa de espectro extendido (BLEE) tipo CTX-M-9. MÉTODOS: Se realizó un estudio retrospectivo de las características clínicas y epidemiológicas del brote causado por E. cloacae complex. La identificación y estudio de sensibilidad de las cepas fueron realizados mediante el sistema semiautomático BD Phoenix™, y la caracterización de la BLEE, por PCR y secuenciación. La tipificación molecular se realizó mediante electroforesis en gel de campo pulsado (PFGE). RESULTADOS: Durante febrero de 2014, 6 pacientes (50% mujeres; media de edad: 61,5 años; rango de edad: 44-76 años) ingresados en la UCI del Complejo Hospitalario de Pontevedra (CHOP) presentaron aislamientos de E. cloacae complex resistente a cefalosporinas de amplio espectro. Tres pacientes desarrollaron infección; uno presentó bacteriemia primaria y shock séptico, y 2 neumonía asociada a ventilación mecánica. En los 3 casos restantes los aislamientos de E. cloacae complex se consideraron colonización. El análisis fenotípico y genotípico reveló que todos los aislados presentaban el mismo perfil por PFGE y que portaban la misma BLEE del tipo CTX-M-9. El brote se controló mediante la mejora de las medidas universales y el aislamiento de contacto de los pacientes infectados y/o colonizados. CONCLUSIÓN: Se describe desde un punto de vista clínico y epidemiológico un brote de E. cloacae complex portador de CTX-M-9 en una UCI

INTRODUCTION: Clinical and epidemiological description of an outbreak in an intensive care unit (ICU) caused by a strain of multidrug-resistant Enterobacter cloacae complex carrying a CTX-M-9-type extended-spectrum beta-lactamase (ESBL). METHODS: A retrospective study of the clinical and epidemiological features of the outbreak caused by E. cloacaecomplex was performed. Identifying and studying the sensitivity of the strains were performed using the semi-automated system BD Phoenix™, and the characterisation of ESBL using PCR and sequencing. Molecular typing was performed by pulsed-field gel electrophoresis (PFGE). RESULTS: During February 2014, 6 patients (50% women; mean age: 61.5 years; age range: 44-76 years) admitted to the ICU of the Hospital of Pontevedra (CHOP) presented resistant E. cloacae complex isolates to extended-spectrum cephalosporins. Three patients developed infection; one had primary bacteraemia and septic shock, and 2 with ventilator-associated pneumonia. In the remaining three cases E. cloacae complex isolates were considered as colonisation. Phenotypic and genotypic analysis revealed that all isolates had the same PFGE profile and carried the same CTX-M-9 ESBL. The outbreak was controlled by improving universal precautions and contact isolation of patients infected and/or colonized. CONCLUSION: The clinical and epidemiological features of an outbreak in an ICU caused by E. cloacae complex carrying CTX-M-9 are described