Impacts of PI3K/protein kinase B pathway activation in reactive astrocytes: from detrimental effects to protective functions.

Astrocytes are the most abundant type of glial cell in the central nervous system. Upon injury and inflammation, astrocytes become reactive and undergo morphological and functional changes. Depending on their phenotypic classification as A1 or A2, reactive astrocytes contribute to both neurotoxic and neuroprotective responses, respectively. However, this binary classification does not fully capture the diversity of astrocyte responses observed across different diseases and injuries. Transcriptomic analysis has revealed that reactive astrocytes have a complex landscape of gene expression profiles, which emphasizes the heterogeneous nature of their reactivity. Astrocytes actively participate in regulating central nervous system inflammation by interacting with microglia and other cell types, releasing cytokines, and influencing the immune response. The phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway is a central player in astrocyte reactivity and impacts various aspects of astrocyte behavior, as evidenced by in silico , in vitro , and in vivo results. In astrocytes, inflammatory cues trigger a cascade of molecular events, where nuclear factor-κB serves as a central mediator of the pro-inflammatory responses. Here, we review the heterogeneity of reactive astrocytes and the molecular mechanisms underlying their activation. We highlight the involvement of various signaling pathways that regulate astrocyte reactivity, including the PI3K/AKT/mammalian target of rapamycin (mTOR), α v ß 3 integrin/PI3K/AKT/connexin 43, and Notch/PI3K/AKT pathways. While targeting the inactivation of the PI3K/AKT cellular signaling pathway to control reactive astrocytes and prevent central nervous system damage, evidence suggests that activating this pathway could also yield beneficial outcomes. This dual function of the PI3K/AKT pathway underscores its complexity in astrocyte reactivity and brain function modulation. The review emphasizes the importance of employing astrocyte-exclusive models to understand their functions accurately and these models are essential for clarifying astrocyte behavior. The findings should then be validated using in vivo models to ensure real-life relevance. The review also highlights the significance of PI3K/AKT pathway modulation in preventing central nervous system damage, although further studies are required to fully comprehend its role due to varying factors such as different cell types, astrocyte responses to inflammation, and disease contexts. Specific strategies are clearly necessary to address these variables effectively.

Body mass index-adjusted calf circumference is associated with mortality in hospitalized older patients with excess weight.

OBJECTIVES: Given the innovative nature of the method, our study aimed to assess the prognostic significance of body mass index (BMI)-adjusted calf circumference (CC) in older patients who are hospitalized. METHODS: This was a unique analysis as part of other cohorts comprising general hospitalized patients aged 60 years or older of both sexes. Only patients with excess weight (BMI ≥ 25 kg/m2) were included. CC was adjusted by reducing 3, 7, or 12 cm for BMI (in kg/m2) within 25-29.9, 30-39.9, and ≥40 kg/m2, respectively. CC was considered low if ≤ 34 cm for males and ≤ 33 cm for females. Clinical outcomes included prolonged length of hospital stay (LOS) and mortality. RESULTS: A total of 222 patients were included. After BMI adjustments, 72.1% of the patients were reclassified from a normal CC category to a low CC category. The frequency of low CC increased from 33.8% to 81.9% following BMI adjustments. Among those reclassified to the low CC, 11 died, compared to only 2 patients in the group that maintained a normal CC classification. BMI-adjusted CC was inversely associated with mortality (HR adjusted 0.84, 95% CI 0.73 to 0.95), but not with prolonged LOS. CONCLUSIONS: Our novel study highlights the prognostic value of BMI-adjusted CC. As an anthropometric marker of muscle mass, it proved to be a predictor of mortality in older patients with high BMI. This adjustment is further important because it may help to better detect low muscle mass in these patients where such conditions might be masked.

Multicenter external validation of prediction models for clinical outcomes after spinal fusion for lumbar degenerative disease.

BACKGROUND: Clinical prediction models (CPM), such as the SCOAP-CERTAIN tool, can be utilized to enhance decision-making for lumbar spinal fusion surgery by providing quantitative estimates of outcomes, aiding surgeons in assessing potential benefits and risks for each individual patient. External validation is crucial in CPM to assess generalizability beyond the initial dataset. This ensures performance in diverse populations, reliability and real-world applicability of the results. Therefore, we externally validated the tool for predictability of improvement in oswestry disability index (ODI), back and leg pain (BP, LP). METHODS: Prospective and retrospective data from multicenter registry was obtained. As outcome measure minimum clinically important change was chosen for ODI with ≥ 15-point and ≥ 2-point reduction for numeric rating scales (NRS) for BP and LP 12 months after lumbar fusion for degenerative disease. We externally validate this tool by calculating discrimination and calibration metrics such as intercept, slope, Brier Score, expected/observed ratio, Hosmer-Lemeshow (HL), AUC, sensitivity and specificity. RESULTS: We included 1115 patients, average age 60.8 ± 12.5 years. For 12-month ODI, area-under-the-curve (AUC) was 0.70, the calibration intercept and slope were 1.01 and 0.84, respectively. For NRS BP, AUC was 0.72, with calibration intercept of 0.97 and slope of 0.87. For NRS LP, AUC was 0.70, with calibration intercept of 0.04 and slope of 0.72. Sensitivity ranged from 0.63 to 0.96, while specificity ranged from 0.15 to 0.68. Lack of fit was found for all three models based on HL testing. CONCLUSIONS: Utilizing data from a multinational registry, we externally validate the SCOAP-CERTAIN prediction tool. The model demonstrated fair discrimination and calibration of predicted probabilities, necessitating caution in applying it in clinical practice. We suggest that future CPMs focus on predicting longer-term prognosis for this patient population, emphasizing the significance of robust calibration and thorough reporting.

Development of full-body rhythmic synchronization in middle childhood.

Rhythmic entrainment is a fundamental aspect of musical behavior, but the skills required to accurately synchronize movement to the beat seem to develop over many years. Motion capture studies of corporeal synchronization have shown immature abilities to lock in to the beat in children before age 5, and reliable synchronization ability in adults without musical training; yet there is a lack of data on full-body synchronization skills between early childhood and adulthood. To document typical rhythmic synchronization during middle childhood, we used a wireless motion capture device to measure period- and phase-locking of full body movement to rhythm and metronome stimuli in 6 to 11 year-old children in comparison with adult data. Results show a gradual improvement with age; however children's performance did not reach adult levels by age 12, suggesting that these skills continue to develop during adolescence. Our results suggest that in the absence of specific music training, full-body rhythmic entrainment skills improve gradually during middle childhood, and provide metrics for examining the continued maturation of these skills during adolescence.

Anterior Cord Syndrome Due to Spontaneous Spinal Epidural Hematoma.

Spontaneous spinal epidural hematoma (SSEH) represents a rare clinical entity with an indeterminate etiology. Timely diagnosis and intervention are imperative due to the significant risk of permanent neurological deficits in the absence of appropriate treatment. This case report presents an instance of SSEH with no clear etiology. The patient arrived at the emergency department with paraplegia, urinary and fecal incontinence, and loss of pain and temperature sensation. She reported that these symptoms began abruptly after sneezing. The patient denied any pertinent medical history or family history. The patient initially experienced epigastric pain, which progressed to paresthesia. Magnetic resonance imaging confirmed an epidural hematoma extending from T2 to T8, necessitating immediate neurosurgical intervention. Although the patient was expected to recover within 72 hours postoperation, her symptoms persisted. Based on her clinical presentation, a diagnosis of anterior cord syndrome secondary to SSEH was confirmed.

Photoacoustic lifetime oxygen imaging of radiotherapy-induced tumor reoxygenation In Vivo.

Purpose: Early detection and diagnosis of cancer is critical for achieving positive therapeutic outcomes. Biomarkers that can provide clinicians with clues to the outcome of a given therapeutic course are highly desired. Oxygen is a small molecule that is nearly universally present in biological tissues and plays a critical role in the effectiveness of radiotherapies by reacting with DNA radicals and subsequently impairing cellular repair of double strand breaks.Techniques for measuring oxygen in biological tissues often use blood oxygen saturation to approximate the oxygen partial pressure in surrounding tissues despite the complex, nonlinear, and dynamic relationship between these two separate oxygen populations. Methods and materials: We combined a directly oxygen-sensitive, tumor-targeted, chemical contrast nanoelement with the photoacoustic lifetime-based (PALT) oxygen imaging technique to obtain image maps of oxygen in breast cancer tumors in vivo. The oxygen levels of patient-derived xenografts in a mouse model were characterized before and after a course of radiotherapy. Results: We show that, independent of tumor size, radiotherapy induced an increase in the overall oxygenation levels of the tumor. Further, this increase in the oxygenation of the tumor significantly correlated with a positive response to radiotherapy, as demonstrated by a reduction in tumor volume over the twenty-day monitoring period following therapy and histological staining. Conclusion: Our PALT imaging presented here is simple, fast, and non-invasive. Facilized by the PALT approach, imaging of tumor reoxygenation may be utilized as a simple, early indicator for evaluating cancer response to radiotherapy. Further characterization of the reoxygenation degree, temporal onset, and possible theragnostic implications are warranted.

Assessment and management of post-surgical pain in adult patients undergoing thoracic surgery: a best practice implementation project.

INTRODUCTION AND OBJECTIVES: Effectiv. e management of post-operative pain improves the condition of patients and reduces their hospital stay. This, in turn, has an impact on caregivers, professionals, and institutions and, as such, is considered a primary indicator of quality. The aim of this project was to improve the assessment and management of post-surgical pain in thoracic surgery patients. METHODS: This implementation project was conducted in a thoracic surgery unit of a tertiary hospital in Spain. The project was guided by the JBI Evidence Implementation Framework, which is grounded in an audit, feedback, and re-audit strategy. A baseline audit was conducted with 44 patients, and barriers to best practice were identified. Strategies were then implemented to improve the assessment and management of post-operative pain. Three follow-up audits were performed using nine audit criteria with 34, 40, and 46 patients, respectively. RESULTS: The baseline audit revealed poor compliance with best practices. After implementing strategies to address areas of non-compliance, health education for patients and caregivers improved up to 80%, while the measurement of pain upon admission and post-surgery rose to 91%. However, patients undergoing pre-operative assessment to guide their post-operative pain management at hospital discharge remained below 50%. CONCLUSIONS: Using a methodology to implement best practices, together with clinical audits, improved compliance with the use of validated scales to assess and manage pain. A multidisciplinary approach improves the quality of care received by patients and contributes to their recovery. SPANISH ABSTRACT: http://links.lww.com/IJEBH/A240.

Phase Angle but Not Psoas Muscle Predicts Nutritional Risk and Prognosis in Males with Hepatocellular Carcinoma.

Despite those with hepatocellular carcinoma (HCC) being at increased risk of malnutrition, there is a notable absence of practical approaches for nutritional assessment in clinical practice. We investigated the usefulness of phase angle (PhA) and Total Psoas Area Index (TPAI) for indicating nutritional risk and HCC prognosis. Weight, height, body mass index (BMI), adductor pollicis muscle thickness (APMT), and handgrip strength (HGS) were assessed. The Nutritional Risk Index (NRI) was calculated. Body composition was assessed using bioimpedance spectroscopy and magnetic resonance imaging. The Child-Turcotte-Pugh (CTP) score and Barcelona-Clinic Liver Cancer (BCLC) classification determined the prognosis. Fifty-one males with HCC were enrolled (CTP C = 11.8%). PhA showed a moderate positive correlation with APMT (r = 0.450; p < 0.001) and HGS (r = 0.418; p = 0.002) and a weak positive correlation with TPAI (r = 0.332; p = 0.021). PhA had a strong positive correlation with NRI (r = 0.614; p < 0.001). Mean PhA values were significantly different according to disease severity (CTP C p = 0.001, and BCLC D p = 0.053). TPAI had no significant correlation with HGS, CTP, or BCLC. PhA was a superior approach for predicting nutritional risk and prognosis in HCC than TPAI. Lower PhA is associated with disease progression, lower muscle mass and function, greater severity of nutritional risk, and increased mortality in HCC.

Lifestyle Clusters of Diet Quality, Sleep, and Screen Time and Associations with Weight Status in Children from Madrid City: ENPIMAD Study.

BACKGROUND: Childhood overweight and obesity is a global concern and has increased in Spain over the last decades. Combinations of lifestyle behaviors (i.e., diet, sleep, and sedentarism) are highly related to weight status. Therefore, this study aimed to identify lifestyle patterns among children from Madrid City, and analyze associations with the prevalence of overweight, obesity, and abdominal obesity, considering socio-economic factors. METHODS: A cross-sectional analysis was conducted on 4545 children from the ENPIMAD study with data on diet, sleep, anthropometric, and socio-economic variables. K-means cluster analysis was used to identify lifestyle clusters, and logistic regressions were used to examine the associations between socio-economic indicators and cluster membership, and between clusters and weight status. RESULTS: Findings show three lifestyle clusters (healthy, mixed, and unhealthy), with boys and older children more represented in the unhealthy cluster. Food insecurity and low socio-economic status were associated with unhealthier clusters in boys and girls. Children in unhealthier clusters were more likely to have obesity and abdominal obesity. However, these associations disappeared in girls after controlling for food insecurity. CONCLUSION: These results provide insight into the combination of behaviors and socio-economic factors associated with childhood obesity that may aid in the design of future interventions.

Factors influencing recovery in a pediatric sample with disorders of consciousness: insights from an observational study.

OBJECTIVE: To estimate rates and time to reach emergence of consciousness from vegetative state/unresponsive wakefulness syndrome (VS/UWS), and explore factors associated with improved recovery in children and adolescents with disorders of consciousness (DoC) following severe traumatic and non-traumatic brain injury. METHODS: Analytical, retrospective, cohort study. Clinical records of consecutively referred patients admitted in VS/UWS to a neurological rehabilitation institute in Argentina, between 2005 and 2021 were reviewed. Seventy children and adolescents were included in the analysis. A specialized 12-week rehabilitation program was administered, and emergence was defined by scores ≥44 points on the Western Neuro Sensory Stimulation Profile (WNSSP), sustained for at least 3 weeks on consecutive weekly evaluations. RESULTS: Emergence from VS/UWS to consciousness occurred within 5.4 (SD 2.6) weeks in almost one-third of patients. Multivariate Cox regression analysis showed emergence was significantly lower in patients with hypoxic ischemic encephalopathy compared to patients with other non-traumatic etiologies [HRadj 0.23 (95% CI 0.06-0.89); p = 0.03)]. CONCLUSIONS: Our findings reinforce growing evidence on the impact of etiology on DoC recovery in pediatric populations, ultimately influencing treatment and family-related decisions in child neurorehabilitation.

Integrated impact of multiple body composition parameters on overall survival in gastrointestinal or genitourinary cancers: A descriptive cohort study.

BACKGROUND: This study aimed to evaluate if combining low muscle mass with additional body composition abnormalities, such as myosteatosis or adiposity, could improve survival prediction accuracy in a large cohort of gastrointestinal and genitourinary malignancies. METHODS: In total, 2015 patients with surgically-treated gastrointestinal or genitourinary cancer were retrospectively analyzed. Skeletal muscle index, skeletal muscle radiodensity, and visceral/subcutaneous adipose tissue index were determined. The primary outcome was overall survival determined by hospital records. Multivariate Cox hazard models were used to identify independent predictors for poor survival. C-statistics were assessed to quantify the prognostic capability of the models with or without incorporating body composition parameters. RESULTS: Survival curves were significantly demarcated by all 4 measures. Skeletal muscle radiodensity was associated with non-cancer-related deaths but not with cancer-specific survival. The survival outcome of patients with low skeletal muscle index was poor (5-year OS; 65.2%), especially when present in combination with low skeletal muscle radiodensity (5-year overall survival; 50.2%). All examined body composition parameters were independent predictors of lower overall survival. The model for predicting overall survival without incorporating body composition parameters had a c-index of 0.68 but increased to 0.71 with the inclusion of low skeletal muscle index and 0.72 when incorporating both low skeletal muscle index and low skeletal muscle radiodensity/visceral adipose tissue index/subcutaneous adipose tissue index. CONCLUSION: Patients exhibiting both low skeletal muscle index and other body composition abnormalities, particularly low skeletal muscle radiodensity, had poorer overall survival. Models incorporating multiple body composition prove valuable for mortality prediction in oncology settings.

Efficacy of alendronic acid solution in removal calcium hydroxide from root canals.

Background: Intracanal medication is considered an alternative after instrumentation in the case of pulp necrosis. However, the elimination of this medication plays an important role in the obturation process, which is why various chelating solutions have currently been studied for this purpose. Therefore, this study aimed to analyze the irrigant based on alendronic acid (AA), which contains in its chemical structure functional groups capable of forming complexes with Ca2+ ions. Material and Methods: 90 single-rooted teeth were crowned and standardized to a length of 13 mm. They were instrumented with a progressive K file (Dentsply Maillefer) from #15 to #30. Ca(OH)2 was placed as intracanal medication for 7 days. The roots were randomly divided into the following 4 experimental groups (n=13) according to the irrigant used: 2.25% NaClO, calcium suspension, 0.22% AA, and 10% citric acid; as well as negative (n=5; saline solution) and positive controls (n=5, 17% EDTA). The roots were divided longitudinally and the % of permeable dentinal tubules (% PDT) was determined by thirds (cervical, middle and apical), by analyzing the micrographs obtained by scanning electron microscope (SEM) and the imageJ program. The quantification of the remaining Ca2+ was determined using the Arsenazo III technique. Results: The Kruskal Wallis test was used for the % of permeable dentinal tubules where a significant difference was determined for the different thirds (p<0.005). In the apical third, citric acid and AA irrigants presented a median of 17.71 and 17.51 % PDT respectively. In relation to the quantification of the remaining total calcium, the lowest value was found with AA with a concentration of 4.83 mmol/L. Conclusions: The 0.22% AA solution has the same capacity to remove Ca(OH)2 from the root canal walls as 17% EDTA and 10% citric acid. Key words:Alendronic acid, calcium hydroxide, citric acid, EDTA, irrigant solution,root canal.

Increasing the use of telemedicine: A field experiment.

Patients are reluctant to use telemedicine health services, compared to its substitute in-person visits. One reason is that telemedicine can be accurately evaluated and compared to its substitute only after the product has been adopted and experienced. As such, an intervention that increases the probability of a first experience can have lasting effects. This article reports the results of a randomized field experiment conducted in collaboration with a health insurance company. During the intervention, half of the households out of 3,469 in the sample received periodic e-mails with information about the available services. It effectively increased the take-up and demand for telemedicine. Within the first 8 months of the experiment, patients assigned to the treatment group were 6 percentage points more likely to have used the service at least once (and had about five times the odds of using telemedicine compared to those in the control group). Eight months after the start of the intervention, the number of virtual consultations by the treatment group was six times larger than that of the control group. These results, even if limited by the sample and context in which the intervention took place, provide additional evidence about how information interventions can increase technological take-up within the health sector and could serve as the stepping stone for evaluating the impact of telemedicine on health outcomes causally.

Synthesis of Azo Analogs for Investigating Phosphatidic Acid-Mediated Signaling.

Phosphatidic acid (PA) is a key signaling lipid that plays a crucial role in regulating various cellular processes. Studies have shown that azobenzene-containing PA analogues can be used as an all-chemical strategy for light-mediated control of PA signaling. These photoswitchable lipids offer a solution to the limitations of traditional bulk dosing methods by allowing for light- and shape-dependent interactions with protein effectors and lipid-metabolizing enzymes. This chapter describes how to synthesize AzoPA and dAzoPA.

Diacylglycerol Signaling in Retinal Ganglion Cells.

With impaired retinal ganglion cell (RGC) function and eventual RGC death, there is a heightened risk of experiencing glaucoma-induced blindness or other optic neuropathies. Poor RGC efficiency leads to limited transmission of visual signals between the retina and the brain by RGC axons. Increased focus on studying lipid messengers found in neurons such as endocannabinoids (eCBs) has importance due to their potential axonal pathway regenerative properties. 2-Arachidonoylglycerol (2-AG), a common eCB, is synthesized from an sn-1 hydrolysis reaction between diacylglycerol (DAG) and diacylglycerol lipase (DAGL). Examination of DAG production allows for future downstream analysis in relation to DAGL functionality. Here, we describe protocol guidelines for extracting RGCs from mouse retinas and subsequent mass spectrometry analysis of the DAG content present within the RGCs.

Downregulation of ATP8B2 to Assess Plasmalogen Distribution and Far1 Expression in Primary Trabecular Meshwork Cells.

The trabecular meshwork (TM) from primary open-angle glaucoma (POAG) cases has been found to contain decreased levels of intracellular plasmalogens. Plasmalogens are a subset of lipids involved in diverse cellular processes such as intracellular signaling, membrane asymmetry, and protein regulation. Proper plasmalogen biosynthesis is regulated by rate-limiting enzyme fatty acyl-CoA reductase (Far1). ATPase phospholipid transporting 8B2 (ATP8B2) is a type IV P-type ATPase responsible for the asymmetric distribution of plasmalogens between the intracellular and extracellular leaflets of the plasma membranes. Here we describe the methodology for extraction and culturing of TM cells from corneal tissue and subsequent downregulation of ATP8B2 using siRNA transfection. Further quantification and downstream effects of ATP8B2 gene knockdown will be analyzed utilizing immunoblotting techniques.

Complete versus partial revascularization for older adults with acute coronary syndrome: a meta-analysis and systematic review of randomized and multivariable adjusted data

BACKGROUND Evidence from randomized studies support complete over culprit-only revascularization for patients with acute coronary artery syndrome (ACS) and multivessel coronary artery diseases (MVD). Whether these findings extend to elderly patients, however, has not been thoroughly explored. METHODS We conducted a systematic review and meta-analysis comparing clinical outcomes of elderly individuals (defined as age ≥75 years) with ACS and MVD submitted to complete vs partial-only percutaneous coronary intervention (PCI). PubMed, Embase, and Cochrane were searched. We computed pooled hazard ratios with 95% confidence intervals (CI) to preserve time time-to-event data. RESULTS We included 7 studies, of which 2 were RCT and 5 were multivariable adjusted cohorts, comprising a total 10 147, of whom 43.8% underwent complete revascularization. As compared with partial-only PCI, complete revascularization was associated with a lower all-cause mortality (hazard ratio 0.71; 95% CI 0.60­0.85; P < 0.01), cardiovascular mortality (hazard ratio 0.64; 95% CI 0.52­0.79; P < 0.01), and recurrent myocardial infarction (hazard ratio 0.65; 95% CI 0.50­0.85; P < 0.01). There was no significant difference between groups regarding the risk of revascularizations (hazard ratio 0.80; 95% CI 0.53­1.20; P = 0.28). CONCLUSION Among elderly patients with ACS and multivessel CAD, complete revascularization is associated with a lower risk of all-cause mortality, cardiovascular mortality, and recurrent myocardial infarction.

A Comprehensive Analytical Review of Polyphenols: Evaluating Neuroprotection in Alzheimer's Disease.

Alzheimer's Disease (AD), a prevalent neurodegenerative disorder, is the primary cause of dementia. Despite significant advancements in neuroscience, a definitive cure or treatment for this debilitating disease remains elusive. A notable characteristic of AD is oxidative stress, which has been identified as a potential therapeutic target. Polyphenols, secondary metabolites of plant origin, have attracted attention due to their potent antioxidant properties. Epidemiological studies suggest a correlation between the consumption of polyphenol-rich foods and the prevention of chronic diseases, including neurodegenerative disorders, which underscores the potential of polyphenols as a therapeutic strategy in AD management. Hence, this comprehensive review focuses on the diverse roles of polyphenols in AD, with a particular emphasis on neuroprotective potential. Scopus, ScienceDirect, and Google Scholar were used as leading databases for study selection, from 2018 to late March 2024. Analytical chemistry serves as a crucial tool for characterizing polyphenols, with a nuanced exploration of their extraction methods from various sources, often employing chemometric techniques for a holistic interpretation of the advances in this field. Moreover, this review examines current in vitro and in vivo research, aiming to enhance the understanding of polyphenols' role in AD, and providing valuable insights for forthcoming approaches in this context.

Presence of intracellular bacterial communities in uroepithelial cells, a potential reservoir in symptomatic and non-symptomatic people.

BACKGROUND: Urinary tract infection is one of the most common infections in humans, affecting women in more proportion. The bladder was considered sterile, but it has a urinary microbiome. Moreover, intracellular bacteria (IB) were observed in uroepithelial cells from children and women with urinary tract infections (UTIs). Here, we evaluated the presence of IB in urine from healthy people and patients with UTI symptoms. METHODS: Midstream urine was self-collected from 141 donors, 77 females and 64 males; 72 belonged to the asymptomatic group and 69 were symptomatic. IB was characterized by a culture-dependent technique and visualized by confocal microscopy. Urine was also subjected to the classical uroculture and isolated bacteria were identified by MALDI-TOF. RESULTS: One-hundred and fifteen uroculture were positive. A significant association was observed between the presence of symptoms and IB (P = 0.007). Moreover, a significant association between the presence of IB, symptoms and being female was observed (P = 0.03). From the cases with IB, Escherichia coli was the most frequent microorganism identified (34.7%), followed by Stenotrophomonas maltophilia (14.2%), Staphylococcus spp (14.2%), and Enterococcus faecalis (10.7%). Intracellular E. coli was associated with the symptomatic group (P = 0.02). Most of the intracellular Staphylococcus spp. were recovered from the asymptomatic group (P = 0.006). CONCLUSIONS: Intracellular bacteria are present in patients with UTI but also in asymptomatic people. Here, we report for the first time, the presence of S. maltophilia, Staphylococcus spp., and Enterobacter cloacae as intracellular bacteria in uroepithelial cells. These findings open new insights into the comprehension of urinary tract infections, urinary microbiome and future therapies. Uroculture as the gold standard could not be enough for an accurate diagnosis in recurrent or complicated cases.