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1.
AANA J ; 92(3): 207-210, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38758715

ABSTRACT

The purpose of this study was to investigate whether the combination of intrathecal dexamethasone and dexmedetomidine in combination with bupivacaine in spinal anesthesia is effective for reducing nausea, vomiting, shivering, and pain. A retrospective review of records was used to examine the outcomes of patients undergoing cesarean delivery under spinal anesthesia with dexamethasone, dexmedetomidine, and bupivacaine. The records of 11 consecutive patients who underwent cesarean delivery under spinal anesthesia with intrathecal dexamethasone and dexmedetomidine in combination with bupivacaine were evaluated. Data collected included patient demographics, medications and fluids administered, presence of nausea, vomiting, shivering, intraoperative breakthrough pain, and postoperative pain. There were no reported complications related to the administration of intrathecal dexamethasone and dexmedetomidine in combination with bupivacaine in this case series of patients. No patients required treatment for intraoperative vomiting, shivering, or breakthrough pain. One patient required opioid pain medication postoperatively. This case series demonstrates that the administration of intrathecal dexamethasone and dexmedetomidine in combination with bupivacaine for patients undergoing elective cesarean section appears to be safe and offers some advantages as to traditional methods of intrathecal delivery for this surgical procedure.


Subject(s)
Anesthesia, Spinal , Anesthetics, Local , Bupivacaine , Cesarean Section , Dexamethasone , Dexmedetomidine , Injections, Spinal , Humans , Dexmedetomidine/administration & dosage , Dexamethasone/administration & dosage , Female , Bupivacaine/administration & dosage , Pregnancy , Adult , Retrospective Studies , Anesthetics, Local/administration & dosage , Pain, Postoperative/drug therapy , Pain, Postoperative/prevention & control , Nurse Anesthetists , Anesthesia, Obstetrical , Postoperative Nausea and Vomiting/prevention & control , Postoperative Nausea and Vomiting/chemically induced , Drug Therapy, Combination
3.
PLoS Biol ; 21(11): e3002389, 2023 Nov.
Article in English | MEDLINE | ID: mdl-37983289

ABSTRACT

The meningeal space is a critical brain structure providing immunosurveillance for the central nervous system (CNS), but the impact of infections on the meningeal immune landscape is far from being fully understood. The extracellular protozoan parasite Trypanosoma brucei, which causes human African trypanosomiasis (HAT) or sleeping sickness, accumulates in the meningeal spaces, ultimately inducing severe meningitis and resulting in death if left untreated. Thus, sleeping sickness represents an attractive model to study immunological dynamics in the meninges during infection. Here, by combining single-cell transcriptomics and mass cytometry by time-of-flight (CyTOF) with in vivo interventions, we found that chronic T. brucei infection triggers the development of ectopic lymphoid aggregates (ELAs) in the murine meninges. These infection-induced ELAs were defined by the presence of ER-TR7+ fibroblastic reticular cells, CD21/35+ follicular dendritic cells (FDCs), CXCR5+ PD1+ T follicular helper-like phenotype, GL7+ CD95+ GC-like B cells, and plasmablasts/plasma cells. Furthermore, the B cells found in the infected meninges produced high-affinity autoantibodies able to recognise mouse brain antigens, in a process dependent on LTß signalling. A mid-throughput screening identified several host factors recognised by these autoantibodies, including myelin basic protein (MBP), coinciding with cortical demyelination and brain pathology. In humans, we identified the presence of autoreactive IgG antibodies in the cerebrospinal fluid (CSF) of second stage HAT patients that recognised human brain lysates and MBP, consistent with our findings in experimental infections. Lastly, we found that the pathological B cell responses we observed in the meninges required the presence of T. brucei in the CNS, as suramin treatment before the onset of the CNS stage prevented the accumulation of GL7+ CD95+ GC-like B cells and brain-specific autoantibody deposition. Taken together, our data provide evidence that the meningeal immune response during chronic T. brucei infection results in the acquisition of lymphoid tissue-like properties, broadening our understanding of meningeal immunity in the context of chronic infections. These findings have wider implications for understanding the mechanisms underlying the formation ELAs during chronic inflammation resulting in autoimmunity in mice and humans, as observed in other autoimmune neurodegenerative disorders, including neuropsychiatric lupus and multiple sclerosis.


Subject(s)
Trypanosoma brucei brucei , Trypanosomiasis, African , Humans , Animals , Mice , Persistent Infection , Meninges/metabolism , Lymphoid Tissue/metabolism , Autoantibodies
4.
Nat Commun ; 14(1): 7070, 2023 11 03.
Article in English | MEDLINE | ID: mdl-37923768

ABSTRACT

In the skin, Trypanosoma brucei colonises the subcutaneous white adipose tissue, and is proposed to be competent for forward transmission. The interaction between parasites, adipose tissue, and the local immune system is likely to drive the adipose tissue wasting and weight loss observed in cattle and humans infected with T. brucei. However, mechanistically, events leading to subcutaneous white adipose tissue wasting are not fully understood. Here, using several complementary approaches, including mass cytometry by time of flight, bulk and single cell transcriptomics, and in vivo genetic models, we show that T. brucei infection drives local expansion of several IL-17A-producing cells in the murine WAT, including TH17 and Vγ6+ cells. We also show that global IL-17 deficiency, or deletion of the adipocyte IL-17 receptor protect from infection-induced WAT wasting and weight loss. Unexpectedly, we find that abrogation of adipocyte IL-17 signalling results in a significant accumulation of Dpp4+ Pi16+ interstitial preadipocytes and increased extravascular parasites in the WAT, highlighting a critical role for IL-17 signalling in controlling preadipocyte fate, subcutaneous WAT dynamics, and local parasite burden. Taken together, our study highlights the central role of adipocyte IL-17 signalling in controlling WAT responses to infection, suggesting that adipocytes are critical coordinators of tissue dynamics and immune responses to T. brucei infection.


Subject(s)
Parasites , Trypanosoma brucei brucei , Humans , Mice , Animals , Cattle , Interleukin-17 , Adipose Tissue , Subcutaneous Fat , Adipose Tissue, White , Cachexia
5.
Healthcare (Basel) ; 11(17)2023 Aug 31.
Article in English | MEDLINE | ID: mdl-37685469

ABSTRACT

Secure attachment, developed through consistent relationships with attachment figures in childhood, is a crucial factor in fostering healthy interpersonal relationships and a positive self-perception. Part of the positive parenting approach and the theory of affect is taken as a basis to assess how perceived security, as an indicator of secure attachment, predicts adult self-concept through the mediating effects of resilience and positive self-esteem. A quantitative, cross-sectional, and nonprobabilistic study was conducted with 383 participants. We unveiled significant positive associations between perceived security, self-concept, resilience, and positive self-esteem. Path analysis demonstrated that resilience and positive self-esteem sequentially mediate the relationship between perceived security during childhood and adult self-concept. Findings suggest that the ability to cope with adversity and personal acceptance underlie the impact of perceived security on self-concept. It is recommended to promote family intervention programs framed within positive parenting that focus on fostering secure attachment given its influence on adult life. It is also deemed essential to incorporate the promotion of resilience and self-esteem into programs aimed at youths, and adult role models can enhance their self-perception and resilience in the face of potential effects stemming from insecure parenting practices.

6.
F1000Res ; 12: 437, 2023.
Article in English | MEDLINE | ID: mdl-37588058

ABSTRACT

Background: Sleeping sickness is caused by the extracellular parasite Trypanosoma brucei and is associated with neuroinflammation and neuropsychiatric disorders, including disruption of sleep/wake patterns, and is now recognised as a circadian disorder. Sleeping sickness is traditionally studied using murine models of infection due to the lack of alternative in vitro systems that fully recapitulate the cellular diversity and functionality of the human brain. The aim of this study is to develop a much-needed in vitro system that reduces and replaces live animals for the study of infections in the central nervous system, using sleeping sickness as a model infection. Methods: We developed a co-culture system using induced pluripotent stem cell (iPSC)-derived cortical human brain organoids and the human pathogen T. b. gambiense to model host-pathogen interactions in vitro. Upon co-culture, we analysed the transcriptional responses of the brain organoids to T. b. gambiense over two time points. Results: We detected broad transcriptional changes in brain organoids exposed to T. b. gambiense, mainly associated with innate immune responses, chemotaxis, and blood vessel differentiation compared to untreated organoids. Conclusions: Our co-culture system provides novel, more ethical avenues to study host-pathogen interactions in the brain as alternative models to experimental infections in mice. Although our data support the use of brain organoids to model host-pathogen interactions during T. brucei infection as an alternative to in vivo models, future work is required to increase the complexity of the organoids ( e.g., addition of microglia and vasculature). We envision that the adoption of organoid systems is beneficial to researchers studying mechanisms of brain infection by protozoan parasites. Furthermore, organoid systems have the potential to be used to study other parasites that affect the brain significantly reducing the number of animals undergoing moderate and/or severe protocols associated with the study of neuroinflammation and brain infections.


Subject(s)
Induced Pluripotent Stem Cells , Trypanosomiasis, African , Humans , Animals , Mice , Trypanosoma brucei gambiense , Neuroinflammatory Diseases , Brain , Organoids
7.
Nat Commun ; 14(1): 5279, 2023 08 29.
Article in English | MEDLINE | ID: mdl-37644007

ABSTRACT

African trypanosomes colonise the skin to ensure parasite transmission. However, how the skin responds to trypanosome infection remains unresolved. Here, we investigate the local immune response of the skin in a murine model of infection using spatial and single cell transcriptomics. We detect expansion of dermal IL-17A-producing Vγ6+ cells during infection, which occurs in the subcutaneous adipose tissue. In silico cell-cell communication analysis suggests that subcutaneous interstitial preadipocytes trigger T cell activation via Cd40 and Tnfsf18 signalling, amongst others. In vivo, we observe that female mice deficient for IL-17A-producing Vγ6+ cells show extensive inflammation and limit subcutaneous adipose tissue wasting, independently of parasite burden. Based on these observations, we propose that subcutaneous adipocytes and Vγ6+ cells act in concert to limit skin inflammation and adipose tissue wasting. These studies provide new insights into the role of γδ T cell and subcutaneous adipocytes as homeostatic regulators of skin immunity during chronic infection.


Subject(s)
Dermatitis , Trypanosoma brucei brucei , Female , Animals , Mice , Interleukin-17 , Persistent Infection , Adiposity , Obesity , Cachexia , Inflammation
8.
IDCases ; 32: e01794, 2023.
Article in English | MEDLINE | ID: mdl-37214183

ABSTRACT

Yeast fungi of the genus Trichosporon spp. can colonize the gastrointestinal tract in humans. In recent decades, the pathogenic role of Trichosporon asahii has been increasingly acknowledged especially in the setting of neutropenic patients with hematological malignancies. However, non-neutropenic patients who are immunosuppressed for other reasons are also at risk of developing invasive forms of this mycosis. We present the case of a 62-year-old male, with a history of ulcerative colitis under immunosuppressive treatment and previous exposure to antibiotics for various bacterial infections, who was admitted to the emergency department with a mycotic aneurysm of the abdominal aorta and left common iliac secondary to T. asahii infection. A multidisciplinary approach of the case (both early medical and surgical interventions) allowed the patient's favorable outcome. The patient was followed for more than two years with no evidence of relapse. We postulate that the diagnosis of invasive Trichosporonosis should be considered in patients with inflammatory bowel disease (IBD) under immunosuppressive treatment and with prior exposure to antibiotics.

9.
Comput Biol Med ; 159: 106909, 2023 06.
Article in English | MEDLINE | ID: mdl-37071937

ABSTRACT

Speech imagery has been successfully employed in developing Brain-Computer Interfaces because it is a novel mental strategy that generates brain activity more intuitively than evoked potentials or motor imagery. There are many methods to analyze speech imagery signals, but those based on deep neural networks achieve the best results. However, more research is necessary to understand the properties and features that describe imagined phonemes and words. In this paper, we analyze the statistical properties of speech imagery EEG signals from the KaraOne dataset to design a method that classifies imagined phonemes and words. With this analysis, we propose a Capsule Neural Network that categorizes speech imagery patterns into bilabial, nasal, consonant-vocal, and vowels/iy/ and/uw/. The method is called Capsules for Speech Imagery Analysis (CapsK-SI). The input of CapsK-SI is a set of statistical features of EEG speech imagery signals. The architecture of the Capsule Neural Network is composed of a convolution layer, a primary capsule layer, and a class capsule layer. The average accuracy reached is 90.88%±7 for bilabial, 90.15%±8 for nasal, 94.02%±6 for consonant-vowel, 89.70%±8 for word-phoneme, 94.33%± for/iy/ vowel and, 94.21%±3 for/uw/ vowel detection. Finally, with the activity vectors of the CapsK-SI capsules, we generated brain maps to represent brain activity in the production of bilabial, nasal, and consonant-vocal signals.


Subject(s)
Brain-Computer Interfaces , Speech , Speech/physiology , Capsules , Electroencephalography/methods , Neural Networks, Computer , Brain/physiology , Imagination/physiology , Algorithms
10.
BMC Res Notes ; 16(1): 22, 2023 Feb 28.
Article in English | MEDLINE | ID: mdl-36849996

ABSTRACT

OBJECTIVE: To assess the effectiveness and safety of hydroxychloroquine (HCQ) prophylaxis for the prevention of SARS-CoV-2 infection in healthcare workers (HCW) on duty during the COVID-19 pandemic. RESULTS: A total of 68 HCWs met the eligibility criteria were randomly allocated to receive HCQ (n = 36) or not (n = 32). There were no significant differences between groups in respects to age, gender, or medical history. Eight participants met the primary efficacy endpoint of SAR-CoV-2 infection during the study period; there was no difference in incidence of SARS-CoV-2 infections between both study arms (HCQ: 5 vs Control: 3, p = 0.538). The relative risk of SARS-CoV-2 infection in the HCQ arm was 1.69 compared to the control group (95%CI 0.41-7.11, p = 0.463); due to poor participant accrual, the resulting statistical power of the primary efficacy outcome was 11.54%. No serious adverse events occurred; however, two (2/36, 5.6%) participants no longer wished to participate in the study and withdrew consent due to recurring grade 1 and 2 adverse events. TRIAL REGISTRATION: ClinicalTrials.gov ID: NCT04414241. (Registered on June 4, 2020).


Subject(s)
COVID-19 , Hydroxychloroquine , Humans , Hydroxychloroquine/adverse effects , Pandemics/prevention & control , COVID-19/prevention & control , COVID-19 Drug Treatment , SARS-CoV-2 , Health Personnel
11.
Data Brief ; 46: 108910, 2023 Feb.
Article in English | MEDLINE | ID: mdl-36687159

ABSTRACT

This dataset contains the full results of a survey on mobility patterns after the Covid-19 pandemic. The survey was conducted in the second trimester of 2021 and collected information from 10000 respondents across 20 urban areas. The questions covered demographic and socio-economic characteristics, employment and job related situation, the use of technological alternatives in daily activities, mobility patterns (trip frequency, purpose, destination, mode, level of comfort), and perceptions as regards the usability of each transport option. Particular emphasis was given to the comparison of current activity to that before the pandemic. The survey combined an online (Computer-Assisted Web Interviews, CAWI) approach with telephone (Computer-Assisted Telephone Interviews, CATI) interviews. The sample in each city was representative of the local demographic and socio-economic profile according to age, sex, employment situation, education and urbanization.

12.
Biometals ; 36(3): 463-472, 2023 06.
Article in English | MEDLINE | ID: mdl-36474100

ABSTRACT

Lactoferrin (LF) has in vitro antiviral activity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This study aimed to determine the effect of bovine lactoferrin (bLF) in the prevention of SARS-CoV-2 infection in health care personnel. A randomized, double-blinded, placebo-controlled clinical trial was conducted in two tertiary hospitals that provide care to patients with SARS-CoV-2 infection in Lima, Peru. Daily supplementation with 600 mg of enteral bLF versus placebo for 90 days was compared. Participants were weekly screened for symptoms suggestive of SARS-CoV-2 infection and molecular testing was performed on suspected episodes. A serological test was obtained from all participants at the end of the intervention. The main outcome included symptomatic and asymptomatic cases. A sub-analysis explored the time to symptomatic infection. Secondary outcomes were the severity, frequency, and duration of symptomatic infection. The study was prematurely cancelled due to the availability of vaccines against SARS-CoV-2 in Peru. 209 participants were enrolled and randomized, 104 received bLF and 105 placebo. SARS-CoV-2 infection occurred in 11 (10.6%) participants assigned to bLF and in 9 (8.6%) participants assigned to placebo without significant differences (Incidence Rate Ratio = 1.23, 95%CI 0.51-3.06, p-value = 0.64). There was no significant effect of bLF on time to symptomatic infection (Hazard Ratio = 1.61, 95%CI 0.62-4.19, p-value = 0.3). There were no significant differences in secondary outcomes. A significant effect of bLF in preventing SARS-CoV-2 infection was not proven. Further studies are needed to assess the effect of bLF supplementation on SARS-CoV-2 infection.Clinical trial registration ClinicalTrials.gov Identifier: NCT04526821, https://clinicaltrials.gov/ct2/show/NCT04526821?term=LACTOFERRIN&cond=COVID-19&cntry=PE&city=Lima&draw=2&rank=1 .


Subject(s)
COVID-19 , Lactoferrin , Humans , COVID-19/prevention & control , COVID-19 Vaccines , Delivery of Health Care , Hydroxychloroquine/therapeutic use , Lactoferrin/therapeutic use , SARS-CoV-2
13.
Antibiotics (Basel) ; 13(1)2023 Dec 20.
Article in English | MEDLINE | ID: mdl-38275316

ABSTRACT

Infectious diseases account for nine percent of annual human deaths, and the widespread emergence of antimicrobial resistances threatens to significantly increase this number in the coming decades. The prospect of antimicrobial peptides (AMPs) derived from venomous animals presents an interesting alternative for developing novel active pharmaceutical ingredients (APIs). Small, cationic and amphiphilic peptides were predicted from the venom gland transcriptome of Pamphobeteus verdolaga using a custom database of the arthropod's AMPs. Ninety-four candidates were chemically synthesized and screened against ATCC® strains of Escherichia coli and Staphylococcus aureus. Among them, one AMP, named PvAMP66, showed broad-spectrum antimicrobial properties with selectivity towards Gram-negative bacteria. It also exhibited activity against Pseudomonas aeruginosa, as well as both an ATCC® and a clinically isolated multidrug-resistant (MDR) strain of K. pneumoniae. The scanning electron microscopy analysis revealed that PvAMP66 induced morphological changes of the MDR K. pneumoniae strain suggesting a potential "carpet model" mechanism of action. The isobologram analysis showed an additive interaction between PvAMP66 and gentamicin in inhibiting the growth of MDR K. pneumoniae, leading to a ten-fold reduction in gentamicin's effective concentration. A cytotoxicity against erythrocytes or peripheral blood mononuclear cells was observed at concentrations three to thirteen-fold higher than those exhibited against the evaluated bacterial strains. This evidence suggests that PvAMP66 can serve as a template for the development of AMPs with enhanced activity and deserves further pre-clinical studies as an API in combination therapy.

14.
Article in Spanish | LILACS | ID: biblio-1431753

ABSTRACT

Introducción: La placenta sintetiza y secreta varias hormonas que permiten la regulación del embarazo, el trabajo de parto y la adaptación metabólica materno-fetal. Su comportamiento asociado al tipo de parto puede dar información relevante sobre efectos epigenéticos. Objetivo: Describir el tipo de parto con los niveles de oxitocina, cortisol y hormonas tiroideas en plasma de cordón umbilical al nacer. Método: A 50 mujeres con embarazos principalmente normales se les cuantificaron los niveles neurohormonales en plasma de cordón umbilical, obtenido inmediatamente tras el periodo expulsivo. Los resultados se incorporaron a la base de datos clínicos de cada participante y se analizaron con Stata v.14.0. El protocolo fue aprobado por el comité de ética. Resultados: Hubo 33 partos vaginales (12 espontáneos, 13 acelerados y 8 inducidos) y 17 cesáreas (7 electivas y 10 de urgencia). Se observaron mayores niveles de cortisol en los partos vaginales acelerados; las cesáreas tuvieron menores niveles de cortisol y hormona estimulante de la tiroides. Las intervenciones clínicas, con altos o bajos niveles hormonales, están en directa relación con el tipo de parto. Conclusiones: El cortisol y la hormona estimulante de la tiroides medidos en plasma de cordón umbilical variaron según el tipo de parto. Esto es una primera cuantificación de hormonas en plasma de cordón umbilical y su posible regulación placentaria a propósito del tipo de parto.


Introduction: The placenta synthesizes and secretes several hormones allowing the regulation of pregnancy, labor and maternal-fetal metabolic adaptation. Their behavior associated with the type of delivery, may provide relevant information on epigenetic effects. Objective: To describe the type of delivery with the levels of oxytocin, cortisol and thyroid hormones in umbilical cord plasma at birth. Method: Neurohormonal levels from umbilical cord plasma obtained immediately post expulsion, were quantified in 50 women with mainly normal pregnancies. Results incorporated into the clinical database of each participant, statistically analyzed in Stata v.14.0. Protocol approved by ethics committee. Results: 33 were vaginal deliveries (12 spontaneous, 13 accelerated, 8 induced) and 17 cesarean sections (7 elective and 10 emergency). Higher cortisol levels were observed in accelerated vaginal deliveries, cesarean sections had lower cortisol and thyroid stimulating hormone levels. While clinical interventions, with high or low hormone levels, were related to the type of delivery. Conclusions: Cortisol and thyroid stimulating hormone measured in umbilical cord plasma varied according to the type of delivery. This is a first quantification of hormones in umbilical cord plasma and their possible placental regulation in relation to the type of delivery.


Subject(s)
Humans , Female , Pregnancy , Adult , Placental Hormones/metabolism , Delivery, Obstetric , Fetal Blood/chemistry , Thyroid Hormones/analysis , Umbilical Cord/chemistry , Hydrocortisone/analysis , Oxytocin/analysis , Cesarean Section , Cross-Sectional Studies , Placental Circulation
16.
Nat Commun ; 13(1): 5752, 2022 09 30.
Article in English | MEDLINE | ID: mdl-36180478

ABSTRACT

Human African trypanosomiasis, or sleeping sickness, is caused by the protozoan parasite Trypanosoma brucei and induces profound reactivity of glial cells and neuroinflammation when the parasites colonise the central nervous system. However, the transcriptional and functional responses of the brain to chronic T. brucei infection remain poorly understood. By integrating single cell and spatial transcriptomics of the mouse brain, we identify that glial responses triggered by infection are readily detected in the proximity to the circumventricular organs, including the lateral and 3rd ventricle. This coincides with the spatial localisation of both slender and stumpy forms of T. brucei. Furthermore, in silico predictions and functional validations led us to identify a previously unknown crosstalk between homeostatic microglia and Cd138+ plasma cells mediated by IL-10 and B cell activating factor (BAFF) signalling. This study provides important insights and resources to improve understanding of the molecular and cellular responses in the brain during infection with African trypanosomes.


Subject(s)
Parasites , Trypanosoma brucei brucei , Trypanosomiasis, African , Animals , B-Cell Activating Factor , Brain/parasitology , Humans , Interleukin-10 , Mice , Microglia , Plasma Cells , Transcriptome , Trypanosoma brucei brucei/genetics , Trypanosomiasis, African/parasitology
17.
iScience ; 25(9): 104797, 2022 Sep 16.
Article in English | MEDLINE | ID: mdl-36034214

ABSTRACT

Rapid imaging of large biological tissue specimens such as ultrathick sections of mouse brain cannot easily be performed with a standard microscope. Optical mesoscopy offers a solution, but thus far imaging has been too slow to be useful for routine use. We have developed two different illuminators for light-sheet mesoscopy with the Mesolens and we demonstrate their use in high-speed optical mesoscale imaging of large tissue specimens. The first light-sheet approach uses Gaussian optics and is straightforward to implement. It provides excellent lateral resolution and high-speed imaging, but the axial resolution is poor. The second light-sheet is a more complex Airy light-sheet that provides sub-cellular resolution in three dimensions that is comparable in quality to point-scanning confocal mesoscopy, but the light-sheet method of illuminating the specimen reduces the imaging time by a factor of 14. This creates new possibilities for high-content, higher-throughput optical bioimaging at the mesoscale.

18.
Front Cell Infect Microbiol ; 12: 903957, 2022.
Article in English | MEDLINE | ID: mdl-35774409

ABSTRACT

Human African Trypanosomiasis (HAT) is a disease caused by the extracellular parasite Trypanosoma brucei that affects the central nervous system (CNS) during the chronic stage of the infection, inducing neuroinflammation, coma, and death if left untreated. However, little is known about the structural change happening in the brain as result of the infection. So far, infection-induced neuroinflammation has been observed with conventional methods, such as immunohistochemistry, electron microscopy, and 2-photon microscopy only in small portions of the brain, which may not be representative of the disease. In this paper, we have used a newly-developed light-sheet illuminator to image the level of neuroinflammation in chronically infected mice and compared it to naïve controls. This system was developed for imaging in combination with the Mesolens objective lens, providing fast sub-cellular resolution for tens of mm3-large imaging volumes. The mouse brain specimens were cleared using CUBIC+, followed by antibody staining to locate Glial Fibrillary Acid Protein (GFAP) expressing cells, primarily astrocytes and ependymocytes, used here as a proxy for cell reactivity and gliosis. The large capture volume allowed us to detect GFAP+ cells and spatially resolve the response to T. brucei infection. Based on morphometric analyses and spatial distribution of GFAP+ cells, our data demonstrates a significant increase in cell dendrite branching around the lateral ventricle, as well as dorsal and ventral third ventricles, that are negatively correlated with the branch extension in distal sites from the circumventricular spaces. To our knowledge, this is the first report highlighting the potential of light-sheet mesoscopy to characterise the inflammatory responses of the mouse brain to parasitic infection at the cellular level in intact cleared organs, opening new avenues for the development of new mesoscale imaging techniques for the study of host-pathogen interactions.


Subject(s)
Trypanosoma brucei brucei , Trypanosomiasis, African , Animals , Astrocytes/metabolism , Brain/diagnostic imaging , Brain/metabolism , Host-Pathogen Interactions , Mice , Trypanosomiasis, African/parasitology
19.
Biomicrofluidics ; 16(2): 024108, 2022 Mar.
Article in English | MEDLINE | ID: mdl-35464137

ABSTRACT

Drug-induced liver injury (DILI) results in over 100 000 hospital attendances per year in the UK alone and is a leading cause for the post-marketing withdrawal of new drugs, leading to significant financial losses. MicroRNA-122 (miR-122) has been proposed as a sensitive DILI marker although no commercial applications are available yet. Extracellular blood microRNAs (miRNAs) are promising clinical biomarkers but their measurement at point of care remains time-consuming, technically challenging, and expensive. For circulating miRNA to have an impact on healthcare, a key challenge to overcome is the development of rapid and reliable low-cost sample preparation. There is an acknowledged issue with miRNA stability in the presence of hemolysis and platelet activation, and no solution has been demonstrated for fast and robust extraction at the site of blood draw. Here, we report a novel microfluidic platform for the extraction of circulating miR-122 from blood enabled by a vertical approach and gravity-based bubble mixing. The performance of this disposable cartridge was verified by standard quantitative polymerase chain reaction analysis on extracted miR-122. The cartridge performed equivalently or better than standard bench extraction kits. The extraction cartridge was combined with electrochemical impedance spectroscopy to detect miR-122 as an initial proof-of-concept toward an application in point-of-care detection. This platform enables the standardization of sample preparation and the detection of miRNAs at the point of blood draw and in resource limited settings and could aid the introduction of miRNA-based assays into routine clinical practice.

20.
Neuropsychobiology ; 81(4): 271-285, 2022.
Article in English | MEDLINE | ID: mdl-35093946

ABSTRACT

INTRODUCTION: Major depressive disorder (MDD) is a prevalent condition which has a well-known association with ischemic cardiomyopathy, probably explained by an inflammatory mediator mechanism. Statins, besides reducing cholesterol production, have pleiotropic effects including anti-inflammatory activity. The goal was to evaluate the effect of statins as an addition to standard therapy on mood status, brain perfusion, and neurocognitive performance in MDD. METHODS: We studied 20 MDD patients with brain single-photon emission tomography and Cambridge Neuropsychological Test Automated Battery (CANTAB), half randomized to 10 mg of Rosuvastatin or placebo, in addition to selective serotonin reuptake inhibitors (SSRIs) therapy and being reevaluated 3 months later. The images were compared using Statistical Parametric Mapping; clinical scores (Hamilton Depression Score with 17 items and Beck's Depression Inventory) as well as neurocognitive parameters were applied as covariances (CoV) to estimate regional cerebral blood flow (rCBF) changes with both therapies. RESULTS: Clinical scores decreased in both groups (p = 0.0001); Beck's presented a larger decrease with statins. We observed significantly rCBF changes expressed as significant larger clusters of voxels (p < 0.05) in the pre/subgenual anterior cingulate plus orbitofrontal cortex and a small area in the posterior cingulate gyrus in the statins group, whereas it was not observed with placebo, when using clinical scores as CoV. A similar pattern of rCBF changes was present with emotions recognition, attentional, paired associates learning, spatial planning, and working memory tasks. CONCLUSION: Short-term use of low-dose statins in MDD patients under SSRIs results in important rCBF changes in key mood associated areas to improvement in neurocognitive performance. These findings, even though demonstrated in a small sample, could open a new therapeutic tool in the comprehensive management of this disorder.


Subject(s)
Depressive Disorder, Major , Hydroxymethylglutaryl-CoA Reductase Inhibitors , Brain/diagnostic imaging , Cerebrovascular Circulation/physiology , Depressive Disorder, Major/diagnostic imaging , Depressive Disorder, Major/drug therapy , Gyrus Cinguli , Humans , Hydroxymethylglutaryl-CoA Reductase Inhibitors/pharmacology , Hydroxymethylglutaryl-CoA Reductase Inhibitors/therapeutic use , Perfusion , Selective Serotonin Reuptake Inhibitors/pharmacology , Selective Serotonin Reuptake Inhibitors/therapeutic use , Tomography, Emission-Computed, Single-Photon/methods
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