Analysis of 16S rRNA genes reveals reduced Fusobacterial community diversity when translocating from saliva to GI sites

Fusobacterium nucleatum is a Gram-negative oral commensal anaerobe which has been increasingly implicated in various gastrointestinal (GI) disorders, including inflammatory bowel disease,appendicitis, GI cancers. The oral cavity harbors a diverse group of Fusobacterium, and it ispostulated that F. nucleatum in the GI tract originate from the mouth. It is not known, however, ifall oral Fusobacterium translocate to the GI sites with equal efficiencies. Therefore, we amplified 16SrRNA genes of F. nucleatum and F. periodonticum, two closely related oral species from matchedsaliva, gastric aspirates, and colon or ileal pouch aspirates of three patients with inflammatorybowel disease (IBD) and three healthy controls, and saliva alone from seven patients with eitheractive IBD or IBD in remission. The 16S rRNA gene amplicons were cloned, and the DNA sequencesdetermined by Sanger sequencing. The results demonstrate that fusobacterial community composition differs more significantly between the oral and GI sites than between different individuals.The oral communities demonstrate the highest level of variation and have the richest pool ofunique sequences, with certain nodes/strains enriched in the GI tract and others diminished duringtranslocation. The gastric and colon/pouch communities exhibit reduced diversity and are moreclosely related, possibly due to selective pressure in the GI tract. This study elucidates selectivetranslocation of oral fusobacteria to the GI tract. Identification of specific transmissible clones willfacilitate risk assessment for developing Fusobacterium-implicated GI disorders.Fil: Richardson, Miles. Columbia University; Estados UnidosFil: Ren, Jihui. Columbia University; Estados UnidosFil: Rubinstein Guichon, Mara Roxana. Columbia University; Estados Unidos. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Taylor, Jamila A.. Columbia University; Estados UnidosFil: Friedman, Richard A.. Columbia University; Estados UnidosFil: Shen, Bo. No especifíca;Fil: Han, Yiping W.. Columbia University; Estados Unido

Analysis of 16S rRNA genes reveals reduced Fusobacterial community diversity when translocating from saliva to GI sites

Fusobacterium nucleatum is a Gram-negative oral commensal anaerobe which has been increasingly implicated in various gastrointestinal (GI) disorders, including inflammatory bowel disease,appendicitis, GI cancers. The oral cavity harbors a diverse group of Fusobacterium, and it ispostulated that F. nucleatum in the GI tract originate from the mouth. It is not known, however, ifall oral Fusobacterium translocate to the GI sites with equal efficiencies. Therefore, we amplified 16SrRNA genes of F. nucleatum and F. periodonticum, two closely related oral species from matchedsaliva, gastric aspirates, and colon or ileal pouch aspirates of three patients with inflammatorybowel disease (IBD) and three healthy controls, and saliva alone from seven patients with eitheractive IBD or IBD in remission. The 16S rRNA gene amplicons were cloned, and the DNA sequencesdetermined by Sanger sequencing. The results demonstrate that fusobacterial community composition differs more significantly between the oral and GI sites than between different individuals.The oral communities demonstrate the highest level of variation and have the richest pool ofunique sequences, with certain nodes/strains enriched in the GI tract and others diminished duringtranslocation. The gastric and colon/pouch communities exhibit reduced diversity and are moreclosely related, possibly due to selective pressure in the GI tract. This study elucidates selectivetranslocation of oral fusobacteria to the GI tract. Identification of specific transmissible clones willfacilitate risk assessment for developing Fusobacterium-implicated GI disorders.Fil: Richardson, Miles. Columbia University; Estados UnidosFil: Ren, Jihui. Columbia University; Estados UnidosFil: Rubinstein Guichon, Mara Roxana. Columbia University; Estados Unidos. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Taylor, Jamila A.. Columbia University; Estados UnidosFil: Friedman, Richard A.. Columbia University; Estados UnidosFil: Shen, Bo. No especifíca;Fil: Han, Yiping W.. Columbia University; Estados Unido

Practical and Ethical Concerns in Implementing Enhanced Surveillance Methods to Improve Continuity of HIV Care: Qualitative Expert Stakeholder Study

BACKGROUND: Retention in HIV care is critical to maintaining viral suppression and preventing further transmission, yet less than 50% of people living with HIV in the United States are engaged in care. All US states have a funding mandate to implement Data-to-Care (D2C) programs, which use surveillance data (eg, laboratory, Medicaid billing) to identify out-of-care HIV-positive persons and relink them to treatment. OBJECTIVE: The purpose of this qualitative study was to identify and describe practical and ethical considerations that arise in planning for and implementing D2C. METHODS: Via purposive sampling, we recruited 43 expert stakeholders-including ethicists, privacy experts, researchers, public health personnel, HIV medical providers, legal experts, and community advocates-to participate in audio-recorded semistructured interviews to share their perspectives on D2C. Interview transcripts were analyzed across a priori and inductively derived thematic categories. RESULTS: Stakeholders reported practical and ethical concerns in seven key domains: permission and consent, government assistance versus overreach, privacy and confidentiality, stigma, HIV exceptionalism, criminalization, and data integrity and sharing. CONCLUSIONS: Participants expressed a great deal of support for D2C, yet also stressed the role of public trust and transparency in addressing the practical and ethical concerns they identified

Expert stakeholders’ perspectives on a Data-to-Care strategy for improving care among HIV-positive individuals incarcerated in jails

Data-to-Care (D2C) uses surveillance data (e.g., laboratory, Medicaid billing) to identify out-of-care HIV-positive persons to re-link them to care. Most US states are implementing D2C, yet few studies have explored stakeholders’ perspectives on D2C, and none have addressed these perspectives in the context of D2C in jail. This article reports findings from qualitative, semi-structured interviews conducted with expert stakeholders regarding their perspectives on the ethical challenges of utilizing D2C to understand and improve continuity of care among individuals incarcerated in jails. Participants included 47 professionals with expertise in ethics and privacy, public health and HIV care, the criminal justice system, and community advocacy. While participants expressed a great deal of support for extending D2C to jails, they also identified many possible risks. Stakeholders discussed many issues specific to D2C in jails, such as heightened stigma in the jail setting, the need for training of jail staff and additional non-medical community-based resources, and the high priority of this vulnerable population. Many experts suggested that the actual likelihood of benefits and harms would depend on contextual details. Implementation of D2C in jails may require novel strategies to minimize risk of disclosing out-of-care patients’ HIV status

Novel Role for the AnxA1-Fpr2/ALX Signaling Axis as a Key Regulator of Platelet Function to Promote Resolution of Inflammation

Background: Ischemia reperfusion injury (I/RI) is a common complication of cardiovascular diseases. Resolution of detrimental I/RI-generated prothrombotic and proinflammatory responses is essential to restore homeostasis. Platelets play a crucial part in the integration of thrombosis and inflammation. Their role as participants in the resolution of thromboinflammation is underappreciated; therefore we used pharmacological and genetic approaches, coupled with murine and clinical samples, to uncover key concepts underlying this role. Methods: Middle cerebral artery occlusion with reperfusion was performed in wild-type or annexin A1 (AnxA1) knockout (AnxA1-/-) mice. Fluorescence intravital microscopy was used to visualize cellular trafficking and to monitor light/dye-induced thrombosis. The mice were treated with vehicle, AnxA1 (3.3 mg/kg), WRW4 (1.8 mg/kg), or all 3, and the effect of AnxA1 was determined in vivo and in vitro. Results: Intravital microscopy revealed heightened platelet adherence and aggregate formation post I/RI, which were further exacerbated in AnxA1-/- mice. AnxA1 administration regulated platelet function directly (eg, via reducing thromboxane B2 and modulating phosphatidylserine expression) to promote cerebral protection post-I/RI and act as an effective preventative strategy for stroke by reducing platelet activation, aggregate formation, and cerebral thrombosis, a prerequisite for ischemic stroke. To translate these findings into a clinical setting, we show that AnxA1 plasma levels are reduced in human and murine stroke and that AnxA1 is able to act on human platelets, suppressing classic thrombin-induced inside-out signaling events (eg, Akt activation, intracellular calcium release, and Ras-associated protein 1 [Rap1] expression) to decrease IIbβ3 activation without altering its surface expression. AnxA1 also selectively modifies cell surface determinants (eg, phosphatidylserine) to promote platelet phagocytosis by neutrophils, thereby driving active resolution. (n=5-13 mice/group or 7-10 humans/group.) Conclusions: AnxA1 affords protection by altering the platelet phenotype in cerebral I/RI from propathogenic to regulatory and reducing the propensity for platelets to aggregate and cause thrombosis by affecting integrin (IIbβ3) activation, a previously unknown phenomenon. Thus, our data reveal a novel multifaceted role for AnxA1 to act both as a therapeutic and a prophylactic drug via its ability to promote endogenous proresolving, antithromboinflammatory circuits in cerebral I/RI. Collectively, these results further advance our knowledge and understanding in the field of platelet and resolution biology.Fil: Senchenkova, Elena Y.. State University of Louisiana; Estados UnidosFil: Ansari, Junaid. State University of Louisiana; Estados UnidosFil: Becker, Felix. University Hospital Muenster; AlemaniaFil: Vital, Shantel A.. State University of Louisiana; Estados UnidosFil: Al-Yafeai, Zaki. State University of Louisiana; Estados UnidosFil: Sparkenbaugh, Erica M.. University North Carolina Chapel Hill; Estados UnidosFil: Pawlinski, Rafal. University North Carolina Chapel Hill; Estados UnidosFil: Stokes, Karen Y.. State University of Louisiana; Estados UnidosFil: Carroll, Jennifer L.. State University of Louisiana; Estados UnidosFil: Dragoi, Ana-Maria. State University of Louisiana; Estados UnidosFil: Qin, Cheng Xue. Baker Heart And Diabetes Institute; AustraliaFil: Ritchie, Rebecca H.. Baker Heart And Diabetes Institute; AustraliaFil: Sun, Hai. University Hospital Muenster; AlemaniaFil: Cuellar-Saenz, Hugo H.. State University of Louisiana; Estados UnidosFil: Rubinstein Guichon, Mara Roxana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas; Argentina. Columbia University; Estados UnidosFil: Han, Yiping W.. Columbia University; Estados UnidosFil: Orr, A. Wayne. University Hospital Muenster; AlemaniaFil: Perretti, Mauro. Queen Mary University Of London; Reino UnidoFil: Granger, D. Neil. State University of Louisiana; Estados UnidosFil: Gavins, Felicity N.E.. State University of Louisiana; Estados Unido

Genetic background and stress influence in the diabetic dysregulation of the immune response. Involvement of oxidative stress

Aunque está clínicamente aceptado que la diabetes predispone a sufrir infecciones severas y los estudios sugieren una asociación entre esta patología y las infecciones, no se conocen los mecanismos que median entre la diabetes y la inmunosupresión. A su vez, el estrés tiene un significativo reconocimiento en el desarrollo y la evolución de la diabetes. Es así que el objetivo del presente trabajo fue estudiar la respuesta inmune en la diabetes mellitus tipo 1, ampliando el conocimiento acerca de los factores genéticos y no genéticos que participan en la evolución del estado diabético y en la alteración de la respuesta inmune. Para esto, se utilizó un modelo experimental murino de diabetes mellitus tipo 1 en dos cepas de ratones: BALB/cByJ y C57Bl/6J y se analizó la respuesta inmune encontrándose que, en los ratones diabéticos de la cepa BALB/cByJ, la repuesta inmune determinada in vivo e in vitro se encontraba afectada, mientras que en los ratones de la cepa C57Bl/6J no lo estaba. A continuación se investigó el efecto del estrés sobre la respuesta inmune en la diabetes. Para esto se utilizó el modelo de estrés crónico moderado (CMS) aplicado luego de la instauración de la diabetes. Los resultados mostraron que en los ratones diabéticos de la cepa BALB/cByJ el CMS provocó alteraciones más tempranas sobre la proliferación linfocitaria probablemente mediadas por el aumento en la glucemia. En estos ratones se observó una correlación positiva entre la glucemia y catecolaminas. En cuanto al efecto sobre la cepa C57Bl/6J, sólo se encontró un aumento en las glucemias sin estar presente la alteración inmune. Dado que la hiperglucemia es el principal factor involucrado en el desarrollo de las complicaciones de la diabetes, se evaluó su participación en los efectos observados. Para esto, se analizó el efecto de la alta glucosa sobre la actividad linfocitaria normal. Las altas concentraciones de glucosa sobre linfocitos provenientes de la cepa BALB/cByJ afectaron directamente la proliferación y condujeron a la muerte de las células, alteración que no se observó en los linfocitos provenientes de la cepa C57Bl/6J. Entre los mecanismos que estarían implicados en los efectos deletéreos de la alta glucosa se destacó el aumento en el estrés oxidativo. Estos resultados señalan la influencia del condicionamiento genético y del estrés en el deterioro de la respuesta inmune en el estado diabético. Si bien la extrapolación de estos resultados a pacientes con diabetes debe ser tomada con precaución, puntualizan la importancia de un temprano y adecuado control de la glucemia como así también de los factores ambientales que puedan alterarla.Diabetes is widely believed to predispose to serious infections. Experimental clinical literature supports an association between diabetes and infection. However, the mechanisms linking diabetes and immunosuppression are not well defined. In particular, stress has a significant recognition in the development and progression of diabetes. Therefore, the aim of the present project was to study the immune response in type 1 diabetes and genetic and nongenetic factors involved in the evolution of the diabetic state and in immune response alteration. For this purpose, an experimental animal model of diabetes was used in two mice strains: BALB/cByJ and C57Bl/6J and the immune response was analyzed. The immune response in BALB/cByJ diabetic mice was affected while in C57Bl/6J it was not. Then, the effect of stress on immune response in the diabetic state was investigated. The animals were exposed to a chronic mild stress model after diabetes induction. Stress exposure in BALB/cByJ mice caused earlier alterations on immune response mediated by the glycaemia increase. Also, a positive correlation between glycaemia and catecholamines was found. In C57Bl/6J mice, stress exposure only caused glycaemia increase without the immune alteration. Hyperglyacemia is the main factor involved in the development of diabetes complication, thus its participation in the observed effects was evaluated. The effect of high glucose incubation on lymphocyte reactivity was studied. In BALB/cByJ lymphocytes, high glucose decreased the proliferation and increased the apoptosis whereas in C57Bl/6J lymphocytes these alterations were not observed. Among the mechanisms involved in the deleterious effects of high glucose, the increase in oxidative stress was the most important. These results show the influence of genetic background and stress in the immune alteration in the diabetic state. Although extrapolation of these results to patients with diabetes should be taken with caution, it is important to emphasize the early and adequate glycaemia control as well as environmental factors too.Fil:Rubinstein Guichon, Mara Roxana. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

Prenatal stress induces up-regulation of glucocorticoid receptors on lymphoid cells modifying the T-cell response after acute stress exposure in the adult life

It has been demonstrated that a short-duration stress (acute stress) may result in immunopreparatory or immunoenhancing physiological conditions. The aim of the present study was to investigate whether exposure to prenatal restraint stress (PRS) influences the impact of acute stress on the T-cell response in the adult life. We found that female mice exposed to PRS (PS mice) did not exhibit changes in the T-cell-dependent IgG antibody production with respect to prenatally non-stressed mice (no-PS mice). However, no-PS mice exposed to acute stress showed an increase of antibody production after antigen stimulation. In contrast, PS mice exhibited a decreased response after an acute situation. Spleen catecholamines and plasma corticosterone levels were increased in acute stress in both PS and no-PS mice. Nevertheless, lymphocyte response to hormones was altered in PS mice. Particularly, inhibitory effect of corticosterone was higher on lymphocytes from PS mice. In addition, an increase in protein levels and mRNA expression of glucocorticoid receptor was found in lymphoid cells from PS mice. These results show that prenatal stress alters the immune intrinsic regulatory mechanism that in turn induces an increased vulnerability to any stressful situation able to modify immune homeostasis.Fil: Pascuan, Cecilia Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Cátedra de Farmacología; ArgentinaFil: Rubinstein Guichon, Mara Roxana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Cátedra de Farmacología; ArgentinaFil: Palumbo, María Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Cátedra de Farmacología; ArgentinaFil: Genaro, Ana Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Cátedra de Farmacología; Argentin

Current understanding of the roles of gut–brain axis in the cognitive deficits caused by perinatal stress exposure

Abstract: Abstract: The term ‘perinatal environment’ refers to the period surrounding birth, which plays a crucial role in brain development. It has been suggested that dynamic communication between the neuro–immune system and gut microbiota is essential in maintaining adequate brain function. This interaction depends on the mother’s status during pregnancy and/or the newborn environment. Here, we show experimental and clinical evidence that indicates that the perinatal period is a critical window in which stress-induced immune activation and altered microbiota compositions produce lasting behavioral consequences, although a clear causative relationship has not yet been established. In addition, we discuss potential early treatments for preventing the deleterious effect of perinatal stress exposure. In this sense, early environmental enrichment exposure (including exercise) and melatonin use in the perinatal period could be valuable in improving the negative consequences of early adversities. The evidence presented in this review encourages the realization of studies investigating the beneficial role of melatonin administration and environmental enrichment exposure in mitigating cognitive alteration in offspring under perinatal stress exposure. On the other hand, direct evidence of microbiota restoration as the main mechanism behind the beneficial effects of this treatment has not been fully demonstrated and should be explored in future studies

Current Understanding of the Roles of Gut–Brain Axis in the Cognitive Deficits Caused by Perinatal Stress Exposure

The term ‘perinatal environment’ refers to the period surrounding birth, which plays a crucial role in brain development. It has been suggested that dynamic communication between the neuro–immune system and gut microbiota is essential in maintaining adequate brain function. This interaction depends on the mother’s status during pregnancy and/or the newborn environment. Here, we show experimental and clinical evidence that indicates that the perinatal period is a critical window in which stress-induced immune activation and altered microbiota compositions produce lasting behavioral consequences, although a clear causative relationship has not yet been established. In addition, we discuss potential early treatments for preventing the deleterious effect of perinatal stress exposure. In this sense, early environmental enrichment exposure (including exercise) and melatonin use in the perinatal period could be valuable in improving the negative consequences of early adversities. The evidence presented in this review encourages the realization of studies investigating the beneficial role of melatonin administration and environmental enrichment exposure in mitigating cognitive alteration in offspring under perinatal stress exposure. On the other hand, direct evidence of microbiota restoration as the main mechanism behind the beneficial effects of this treatment has not been fully demonstrated and should be explored in future studies.Fil: Rubinstein Guichon, Mara Roxana. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Burgueño, Adriana Laura. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Quiroga, Sofía. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Wald, Miriam Ruth. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Genaro, Ana Maria. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; Argentin