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Extended-spectrum ß-lactamase (ESBL) producing Escherichia coli represents a formidable challenge in the field of microbiology and public health due to its resistance to commonly used antibiotics. These strains pose a serious threat to human and animal health, underscoring the urgency of comprehensive research and surveillance. The ongoing investigation seeks ESBL producing E. coli strains from pig farms and slaughterhouses in West Bengal and Assam, India. A total of 309 samples were collected: nasal swabs (25), rectal swabs (25) from healthy pigs, pig pen soil (45), faeces (55), slaughterhouse effluents (115), and cleaning water (44). In these samples, 154 tested positive for E. coli, indicating a 49.8% prevalence. Among 154 E. coli isolates, 23 (14.9%) produced ESBLs, sourced from pig rectal swabs (7.1%), faeces (10.7%), slaughterhouse effluents (26.1%), and cleaning water (11.7%). Significantly, 4 ESBL E. coli isolates (6.6%) exclusively emerged from pig slaughterhouse effluents, displaying imipenem-resistant properties. The majority of ESBL E. coli primarily produced CTX-M and CMY, with consistent genetic markers bla CTX-M (100%) and bla CMY (82.6%). Remarkably, 2 (8.6%) of 17 ESBL E. coli isolates from pig slaughterhouse effluents carried the genetic marker bla NDM1. These findings stress implementing thorough surveillance in pig farms and local slaughterhouses. This proactive approach is crucial to identify ESBL E. coli strains, enhancing public health protection.
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Glaucoma is a neurodegenerative disease affecting millions worldwide, characterised by retinal ganglion cell (RGC) degeneration which leads to blindness in more advanced cases. Although the pathogenesis and underlying mechanisms of glaucoma are not fully understood, there are theories that hint at demyelination playing a role in the disease process. Demyelination, or the degeneration of the myelin sheath surrounding axons, has been found in previous studies using animal models of glaucoma and clinical assessments of glaucoma patients. However, this has not been fully realised or quantified in glaucoma patients. Utilising postmortem optic nerve samples from glaucoma and healthy subjects, various immunohistochemical and morphological assessments were performed to determine the extent, if any, of demyelination in glaucomatous optic nerves. Our findings revealed that alongside nerve shrinkage and degeneration of nerve tissue fascicles, there were significantly less myelin proteins, specifically myelin basic protein (MBP), in glaucoma optic nerves. Additionally, the loss of MBP was correlated with decreased oligodendrocyte (OLG) precursors and increasing glial activity. This further supports previous evidence that demyelination may be a secondary degenerative process associated with glaucoma disease progression. Not only do these results provide evidence for potential disease mechanisms, but this is also the first study to quantify optic nerve demyelination in glaucoma postmortem tissue.
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Cystic fibrosis (CF) is a genetic disorder affecting nearly 105,000 patients worldwide and is characterized by poor respiratory function due to accumulation of thick mucus in the lungs, which not just acts as a physical barrier, but also provides a breeding ground for bacterial infections. These infections can be controlled with the help of antibiotics which can be delivered directly into the lungs for amplifying the local anti-bacterial effect. More than 50 % of CF patients are associated with Pseudomonas aeruginosa infection in their lungs which requires antibiotics such as Aztreonam (AZT). In this study, we prepared inhalable AZT-loaded lipid nanoparticles using Hot-melt extrusion (HME) coupled with probe sonication to target Pseudomonas aeruginosa infection in the lungs. The optimized nanoparticles were tested for physicochemical properties, stability profile, in-vitro aerosolization, and antimicrobial activity against Pseudomonas aeruginosa. The optimized nanoparticles with a PEI concentration of 0.1 % demonstrated a uniform particle size of <50 nm, a spherical shape observed under a transmission electron microscope, and >70 % drug entrapment. Incorporating cationic polymer, PEI, resulted in sustained drug release from the lipid nanoparticles. The in-vitro aerosolization studies exhibited a mass median aerodynamic diameter (MMAD) of <4.3 µm, suggesting deposition of the nanoparticles in the respirable airway. The antimicrobial activity against Pseudomonas aeruginosa showed the minimum inhibitory concentration of the formulation is 2-fold lower than plain AZT. Stability profile showed the formulations are stable after exposure to accelerated conditions. In conclusion, hot-melt extrusion in combination with probe sonication can be used as a potential method for the continuous production of AZT-loaded lipid nanoparticles with enhanced anti-bacterial activity.
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The World Health Organization Classification of Tumors of the Central Nervous System (WHO CNS5) significantly revised the terminology and diagnostic criteria of "mesenchymal non-meningothelial" tumors of CNS to better align with the classification of these soft tissue tumors outside the CNS. The CNS chapter only covers the entities with distinct histological or molecular characteristics that occur exclusively or primarily in the CNS. These tumors usually arise from the meninges and are rarely intraparenchymal in origin, mainly in the supratentorial compartment. These tumors are grouped into three main categories: soft tissue, chondro-osseous, and notochordal. Soft tissue tumors, the largest group, are further divided into fibroblastic, vascular, and skeletal muscle subtypes. Notably, a new subcategory for "tumors of uncertain differentiation" has been introduced, encompassing three new histomolecular entities: FET::CREB fusion-positive, CIC-rearranged sarcoma, and primary intracranial sarcoma, DICER1-mutant. Emerging entities like dural angioleiomyomas and spindle cell neoplasms with NTRK-rearrangements have been reviewed, although not introduced in WHO CNS5. Given the often non-specific histology and immunophenotype of mesenchymal nonmeningothelial tumors of uncertain differentiation, molecular techniques have become indispensable for accurate diagnosis. This review provides a comprehensive overview of primary mesenchymal non-meningothelial CNS tumors, including their clinical, radiological, histopathological, and molecular characteristics and treatment strategies.ABBREVIATIONS: ALK: Anaplastic lymphoma kinase; ATF1: activating transcriptase factor-1; CREB: cAMP response element-binding protein; CREM: cAMP response element modulator; CIC: Capicua transcriptional receptor; EWSR1: Ewing sarcoma RNA binding protein; FUS: fused in sarcoma; NAB2: nerve growth factor-inducible protein A binding protein 2; STAT6: signal transducer and activator of transcription 6; WHO: World Health Organization WHO CNS5: World Health Organization Classification of Tumors of the Central Nervous System, fifth edition.
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Neurodegenerative disorders, including Dementia, Parkinson's disease, various Vision disorders, Multiple sclerosis, and transsynaptic degenerative changes represent a significant challenge in aging populations. This editorial synthesizes and discusses recent advancements in understanding the genetic and environmental factors contributing to these diseases. Central to these advancements is the role of neuroinflammation and oxidative stress, which exacerbate neuronal damage and accelerate disease progression. Emerging research underscores the significance of mitochondrial dysfunction and protein aggregation in neurodegenerative pathology, highlighting shared mechanisms across various disorders. Innovative therapeutic strategies, including gene therapy, CRISPR-Cas technology, and the use of naturally occurring antioxidant molecules, are being investigated to target and manage these conditions. Additionally, lifestyle interventions such as exercise and healthy diet have shown promise in enhancing brain plasticity and reducing neuroinflammation. Advances in neuroimaging and biomarker discovery are necessary to improve early diagnosis, while clinical and preclinical studies are essential for the translation of these novel treatments. This edition aims to bridge the gap between molecular mechanisms and therapeutic applications, offering insights into potential interventions to mitigate the impact of neurodegenerative diseases. By establishing a deeper understanding of these complex processes, we aim to move closer to effective prevention and treatment strategies, ultimately improving the quality of life for those affected by neurodegenerative disorders.
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Envejecimiento , Encéfalo , Enfermedades Neurodegenerativas , Humanos , Enfermedades Neurodegenerativas/terapia , Enfermedades Neurodegenerativas/genética , Enfermedades Neurodegenerativas/metabolismo , Enfermedades Neurodegenerativas/patología , Envejecimiento/patología , Encéfalo/patología , Encéfalo/metabolismo , Inflamación/genética , Inflamación/terapia , Estrés Oxidativo , Enfermedades Neuroinflamatorias/metabolismo , Enfermedades Neuroinflamatorias/genéticaRESUMEN
Glaucoma is a neurodegenerative disease affecting millions worldwide, characterised by retinal ganglion cell (RGC) degeneration which leads to blindness in more advanced cases. Although the pathogenesis and underlying mechanisms of glaucoma are not fully understood, there are theories that hint at demyelination playing a role in the disease process. Demyelination, or the degeneration of the myelin sheath surrounding axons, has been found in previous studies using animal models of glaucoma and clinical assessments of glaucoma patients. However, this has not been fully realised or quantified in glaucoma patients. Utilising postmortem optic nerve samples from glaucoma and healthy subjects, various immunohistochemical and morphological assessments were performed to determine the extent, if any, of demyelination in glaucomatous optic nerves. Our findings revealed that alongside nerve shrinkage and degeneration of nerve tissue fascicles, there were significantly less myelin proteins, specifically myelin basic protein (MBP), in glaucoma optic nerves. Additionally, the loss of MBP was correlated with decreased oligodendrocyte (OLG) precursors and increasing glial activity. This further supports previous evidence that demyelination may be a secondary degenerative process associated with glaucoma disease progression. Not only do these results provide evidence for potential disease mechanisms, but this is also the first study to quantify optic nerve demyelination in glaucoma postmortem tissue.
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Enfermedades Desmielinizantes , Glaucoma , Nervio Óptico , Humanos , Nervio Óptico/patología , Glaucoma/patología , Glaucoma/metabolismo , Anciano , Masculino , Femenino , Enfermedades Desmielinizantes/patología , Persona de Mediana Edad , Autopsia , Anciano de 80 o más Años , Proteína Básica de Mielina/metabolismo , Células Ganglionares de la Retina/patología , Vaina de Mielina/patologíaRESUMEN
Neuropeptide Y (NPY), an endogenous peptide composed of 36 amino acids, has been investigated as a potential therapeutic agent for neurodegenerative diseases due to its neuroprotective attributes. This study investigated the neuroprotective effects of NPY in a mouse model of glaucoma characterized by elevated intraocular pressure (IOP) and progressive retinal ganglion cell degeneration. Elevated IOP in mice was induced through intracameral microbead injections, accompanied by intravitreal administration of NPY peptide. The results demonstrated that NPY treatment preserved both the structural and functional integrity of the inner retina and mitigated axonal damage and degenerative changes in the optic nerve under high IOP conditions. Further, NPY treatment effectively reduced inflammatory glial cell activation, as evidenced by decreased expression of glial fibrillary acidic protein and Iba-1. Notably, endogenous NPY expression and its receptors (NPY-Y1R and NPY-Y4R) levels were negatively affected in the retina under elevated IOP conditions. NPY treatment restored these changes to a significant extent. Molecular analysis revealed that NPY mediates its protective effects through the mitogen-activated protein kinase (MAPK) and PI3K/Akt signaling pathways. These findings highlight the therapeutic potential of NPY in glaucoma treatment, underscoring its capacity to preserve retinal health, modulate receptor expression under stress, reduce neuroinflammation, and impart protection against axonal impairment.
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A left ventricular pseudoaneurysm, an ominous consequence of acute myocardial infarction, poses a significant threat to patient well-being. Prompt and accurate diagnosis is crucial for improving patient outcomes. This report describes diagnostic imaging findings for identifying left ventricular pseudoaneurysms, emphasizing the critical role of cardiac magnetic resonance imaging alongside other modalities. We present two cases of patients with a history of myocardial infarction who presented with palpitations, chest pain, and shortness of breath. Initial 2D echocardiography in both patients revealed aneurysmal dilation of the left ventricle. Cardiac MRI was then performed, confirming the diagnosis in both cases.
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Numerous hydrological applications, such as soil erosion estimation, water resource management, and rain driven damage assessment, demand accurate and reliable rainfall erosivity data. However, the scarcity of gauge rainfall records and the inherent uncertainty in satellite and reanalysis-based rainfall datasets limit rainfall erosivity assessment globally. Here, we present a new global rainfall erosivity dataset (0.1° × 0.1° spatial resolution) integrating satellite (CMORPH and IMERG) and reanalysis (ERA5-Land) derived rainfall erosivity estimates with gauge rainfall erosivity observations collected from approximately 6,200 locations across the globe. We used a machine learning-based Gaussian Process Regression (GPR) model to assimilate multi-source rainfall erosivity estimates alongside geoclimatic covariates to prepare a unified high-resolution mean annual rainfall erosivity product. It has been shown that the proposed rainfall erosivity product performs well during cross-validation with gauge records and inter-comparison with the existing global rainfall erosivity datasets. Furthermore, this dataset offers a new global rainfall erosivity perspective, addressing the limitations of existing datasets and facilitating large-scale hydrological modelling and soil erosion assessments.
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Neurodegenerative and demyelinating disease, such as multiple sclerosis (MS) are at the forefront of medical research and the discovery of new drugs and therapeutics. One phenomenon of degeneration seen in these diseases is transsynaptic degeneration (TSD), where damage from one axon spreads to the other axons that are connected to it synaptically. It has previously been found that demyelination occurs prior to neuronal loss in an experimental form of induced TSD. Retinoid-x receptor (RXR) agonists have been shown to promote remyelination. Therefore, this study aimed to reveal the effects of a novel endogenous RXR-γ agonist, 9-cis-13,14-dihydroretinoic acid (9CDHRA), on preventing or restoring the effects of TSD. 9CDHRA was administered to mice following optic nerve crush (ONC) procedures, and electrophysiology (visual evoked potential, VEP) and histological (immunofluorescent) assessments were performed. It was found that 9CDHRA treatment effectively delayed glial activation and reduced the presence of apoptosis at the site of injury and further anterogradely in the visual system, including the lateral geniculate nucleus (LGN) and primary visual cortex (V1). Most notably, 9CDHRA was able to maintain myelin levels following ONC, and effectively protected from demyelination. This was corroborated by VEP recordings with improved P1 latency. The promising findings regarding the injury attenuating and myelin protecting properties of 9CDHRA necessitates further investigations into the potential therapeutic uses of this compound.
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Enfermedades Desmielinizantes , Modelos Animales de Enfermedad , Ratones Endogámicos C57BL , Animales , Enfermedades Desmielinizantes/tratamiento farmacológico , Enfermedades Desmielinizantes/patología , Tretinoina/farmacología , Fármacos Neuroprotectores/farmacología , Potenciales Evocados Visuales/efectos de los fármacos , Potenciales Evocados Visuales/fisiología , Masculino , Receptores X Retinoide/agonistas , Receptores X Retinoide/metabolismo , Degeneración Nerviosa/tratamiento farmacológico , Degeneración Nerviosa/patología , Ratones , Nervio Óptico/efectos de los fármacos , Nervio Óptico/patología , Apoptosis/efectos de los fármacosRESUMEN
BACKGROUND: Direct common carotid puncture (DCP) is conventionally used as a bailout technique in stroke patients. However, little is known about the relevant anatomy. Our objective was to examine the relationship of the common carotid artery (CCA) to surrounding structures based on different DCP trajectories passing through the artery's center. METHODS: Fifty randomly selected head/neck CTAs were analyzed. The trajectory of DCP and relationship to the internal jugular vein (IJV) and thyroid were analyzed at 1 cm intervals above the clavicle on 7 axial sections. Using the trans-carotid sagittal plane as the 0° trajectory, we plotted 3 additional trajectories at 30° intervals and the relationship with the IJV and thyroid proximity was graded as following: 0=absent, 1=adjacent, and 2=crossing. The CCA tortuosity index was also analyzed for each vessel. RESULTS: Analysis of 2800 trajectories across 100 CCAs showed that the IJV and thyroid were least encountered on the axial sections 2 cm above the clavicle, at 0° on the right (9 thyroids and 6 IJV), and at 90° on the left (0 Thyroids and 14 IJVs). The tortuosity index of the CCA was significantly lower above the clavicle than its entire length (P < 0.001). CONCLUSIONS: DCP performed 2 cm above the clavicle at 0° on the right, and 90° on the left appears to minimize encounters with the IJV and thyroid gland, reducing potential complications. However, despite these findings, ultrasound guidance remains vital for DCP safety. Further focus on endovascular device safety in DCP is needed.
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Arteria Carótida Común , Venas Yugulares , Punciones , Humanos , Arteria Carótida Común/anatomía & histología , Arteria Carótida Común/diagnóstico por imagen , Masculino , Femenino , Anciano , Punciones/métodos , Persona de Mediana Edad , Venas Yugulares/anatomía & histología , Venas Yugulares/diagnóstico por imagen , Glándula Tiroides/anatomía & histología , Glándula Tiroides/diagnóstico por imagen , Glándula Tiroides/irrigación sanguínea , Anciano de 80 o más Años , Angiografía por Tomografía Computarizada , Adulto , Accidente Cerebrovascular/diagnóstico por imagenRESUMEN
Extracellular vesicles (EVs) are nanosized vesicles with a lipid bilayer that are secreted by cells and play a critical role in cell-to-cell communication. Despite the promising reports regarding their diagnostic and therapeutic potential, the utilization of EVs in the clinical setting is limited due to insufficient information about their cargo and a lack of standardization in isolation and analysis methods. Considering protein cargos in EVs as key contributors to their therapeutic potency, we conducted a tandem mass tag (TMT) quantitative proteomics analysis of three subpopulations of mesenchymal stem cell (MSC)-derived EVs obtained through three different isolation techniques: ultracentrifugation (UC), high-speed centrifugation (HS), and ultracentrifugation on sucrose cushion (SU). Subsequently, we checked EV marker expression, size distribution, and morphological characterization, followed by bioinformatic analysis. The bioinformatic analysis of the proteome results revealed that these subpopulations exhibit distinct molecular and functional characteristics. The choice of isolation method impacts the proteome of isolated EVs by isolating different subpopulations of EVs. Specifically, EVs isolated through the high-speed centrifugation (HS) method exhibited a higher abundance of ribosomal and mitochondrial proteins. Functional apoptosis assays comparing isolated mitochondria with EVs isolated through different methods revealed that HS-EVs, but not other EVs, induced early apoptosis in cancer cells. On the other hand, EVs isolated using the sucrose cushion (SU) and ultracentrifugation (UC) methods demonstrated a higher abundance of proteins primarily involved in the immune response, cell-cell interactions and extracellular matrix interactions. Our analyses unveil notable disparities in proteins and associated biological functions among EV subpopulations, underscoring the importance of meticulously selecting isolation methods and resultant EV subpopulations based on the intended application.
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Owing to the lack of effective vaccines, current control measures and eradication strategies for the African swine fever virus (ASFV) rely on early detection and stringent stamping-out procedures. In the present study, we developed two independent isothermal amplification assays, namely, loop-mediated isothermal amplification (LAMP) and polymerase spiral reaction (PSR), for quick visualization of the ASFV genome in clinical samples. Additionally, a quantitative real-time PCR (qRT-PCR)-based hydrolysis probe assay was developed for comparative assessment of sensitivity with the developed isothermal assays. The analytical sensitivity of the LAMP, PSR, and qRT-PCR was found to be 2.64 ×105 copies/µL, 2.64 ×102 copies/µL, and 2.64 ×101 copies/µL, respectively. A total of 165 clinical samples was tested using the developed visual assays. The relative accuracy, relative specificity, and relative diagnostic sensitivity for LAMP vs PSR were found to be 95.37% vs 102.48%, 97.46% vs 101.36%, and 73.33% vs 113.33%, respectively.
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Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Técnicas de Amplificación de Ácido Nucleico , Sensibilidad y Especificidad , Virus de la Fiebre Porcina Africana/genética , Virus de la Fiebre Porcina Africana/aislamiento & purificación , Animales , Técnicas de Amplificación de Ácido Nucleico/métodos , Porcinos , Fiebre Porcina Africana/diagnóstico , Fiebre Porcina Africana/virología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Técnicas de Diagnóstico Molecular/métodos , Genoma Viral/genéticaRESUMEN
Escherichia coli and Staphylococcus aureus are the most important food borne pathogen transmitting from animal meat and meat products. Therefore, it is vital to design an accurate and specific diagnostic tool for identifying those food-borne pathogens in animal meat and meat products. In the current study, E. coli, methicillin-resistant and sensitive S. aureus (MRSA and MSSA) were simultaneously detected using a developed triplex PCR-based technique. To obtain an optimal reaction parameter, the multiplex assay was optimised by changing just one parameter while holding the others constant. Specificity of the assay was assessed using several porcine bacterial template DNA. The plasmid DNA was used to test the multiplex PCR assay's sensitivity and interference in spiked pork samples. E. coli, MRSA, and MSSA each have PCR amplified products with sizes of 335, 533, and 209 bp, respectively. The assay detects a minimum microbial load of 102 CFU/µl for all the three pathogens and can identify bacterial DNA as low as 10-2 ng/µl. The assay was validated employing 210 pork samples obtained from retail meat shops and slaughter houses, with MRSA, E. coli, and MSSA with the occurrence rate of 1.9%, 42.38%, and 18.1%, respectively. The rate of mixed bacterial contamination in pork meat samples examined with the developed method was 6.19%, 1.43%, 1.90%, and 1.43% for MSSA & E. coli, MRSA & E. coli, MSSA & MRSA, and E. coli, MSSA & MRSA, respectively. The developed multiplex PCR assay is quick and efficient, and it can distinguish between different bacterial pathogens in a single reaction tube.
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Pig posture is closely linked with livestock health and welfare. There has been significant interest among researchers in using deep learning techniques for pig posture detection. However, this task is challenging due to variations in image angles and times, as well as the presence of multiple pigs in a single image. In this study, we explore an object detection and segmentation algorithm based on instance segmentation scoring to detect different pig postures (sternal lying, lateral lying, walking, and sitting) and segment pig areas in group images, thereby enabling the identification of individual pig postures within a group. The algorithm combines a residual network with 50 layers and a feature pyramid network to extract feature maps from input images. These feature maps are then used to generate regions of interest (RoI) using a region candidate network. For each RoI, the algorithm performs regression to determine the location, classification, and segmentation of each pig posture. To address challenges such as missing targets and error detections among overlapping pigs in group housing, non-maximum suppression (NMS) is used with a threshold of 0.7. Through extensive hyperparameter analysis, a learning rate of 0.01, a batch size of 512, and 4 images per batch offer superior performance, with accuracy surpassing 96%. Similarly, the mean average precision (mAP) exceeds 83% for object detection and instance segmentation under these settings. Additionally, we compare the method with the faster R-CNN object detection model. Further, execution times on different processing units considering various hyperparameters and iterations have been analyzed.
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Algoritmos , Aprendizaje Profundo , Vivienda para Animales , Procesamiento de Imagen Asistido por Computador , Postura , Animales , Porcinos , Procesamiento de Imagen Asistido por Computador/métodosRESUMEN
Outcomes of patients with hematologic malignancies requiring ICU care for critical illness are suboptimal and represent a major unmet need in this population. We present data from a dedicated haematology oncology setting including 63 patients with a median age of 60 years admitted to the ICU for critical illness with organ dysfunction. The most common underlying diagnosis was multiple myeloma (30%) followed by acute myeloid leukemia (25%). Chemotherapy had been initiated for 90.7% patients before ICU admission. The most common indication for ICU care was respiratory failure (36.5%) and shock (17.5%) patients. Evidence of sepsis was present in 44 (69%) patients. After shifting to ICU, 32 (50%) patients required inotropic support and 18 (28%) required invasive mechanical ventilation. After a median of 5 days of ICU stay, 43.1% patients had died, most commonly due to multiorgan dysfunction. Risk of mortality was higher with involvement of more than two major organs (p = .001), underlying AML (p = .001), need for mechanical ventilation (p = .001) and high inotrope usage (p = .004). Neutropenia was not associated with mortality. Our study indicates high rates of short term mortality and defines prognostic factors which can be used to prognosticate patients and establish goals of care. Supplementary Information: The online version contains supplementary material available at 10.1007/s12288-024-01757-3.
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Bifurcations are a common site for saccular aneurysms, but rarely can be a site for dissecting aneurysms. Identification of these aneurysms is extremely important because the management plan depends on it. We describe a rare case of a ruptured dissecting aneurysm at the right ICA bifurcation in a pre-teen child which posed a diagnostic dilemma but ultimately was successfully managed with flow diversion.
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Disección Aórtica , Humanos , Diagnóstico Diferencial , Disección Aórtica/diagnóstico por imagen , Disección Aórtica/cirugía , Aneurisma Intracraneal/diagnóstico por imagen , Aneurisma Intracraneal/cirugía , Masculino , Angiografía Cerebral , Niño , Aneurisma Roto/diagnóstico por imagen , Aneurisma Roto/cirugía , Disección de la Arteria Carótida Interna/diagnóstico por imagen , Resultado del TratamientoRESUMEN
Viral infection disrupts the normal regulation of the host gene's expression. In order to normalise the expression of dysregulated host genes upon virus infection, analysis of stable reference housekeeping genes using quantitative real-time-PCR (qRT-PCR) is necessary. In the present study, healthy and African swine fever virus (ASFV) infected porcine tissues were assessed for the expression stability of five widely used housekeeping genes (HPRT1, B2M, 18 S rRNA, PGK1 and H3F3A) as reference genes using standard algorithm. Total RNA from each tissue sample (lymph node, spleen, kidney, heart and liver) from healthy and ASFV-infected pigs was extracted and subsequently cDNA was synthesized, and subjected to qRT-PCR. Stability analysis of reference genes expression was performed using the Comparative delta CT, geNorm, BestKeeper and NormFinder algorithm available at RefFinder for the different groups. Direct Cycle threshold (CT) values of samples were used as an input for the web-based tool RefFinder. HPRT1 in spleen, 18 S rRNA in liver and kidney and H3F3A in heart and lymph nodes were found to be stable in the individual healthy tissue group (group A). The majority of the ASFV-infected organs (liver, kidney, heart, lymph node) exhibited H3F3A as stable reference gene with the exception of the ASFV-infected spleen, where HPRT1 was found to be the stable gene (group B). HPRT1 was found to be stable in all combinations of all CT values of both healthy and ASFV-infected porcine tissues (group C). Of five different reference genes investigated for their stability in qPCR analysis, the present study revealed that the 18 S rRNA, H3F3A and HPRT1 genes were optimal reference genes in healthy and ASFV-infected different porcine tissue samples. The study revealed the stable reference genes found in healthy as well as ASF-infected pigs and these reference genes identified through this study will form the baseline data which will be very useful in future investigations on gene expression in ASFV-infected pigs.
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Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Reacción en Cadena en Tiempo Real de la Polimerasa , Estándares de Referencia , Animales , Fiebre Porcina Africana/virología , Porcinos , Virus de la Fiebre Porcina Africana/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/normas , Perfilación de la Expresión Génica , Genes Esenciales/genéticaRESUMEN
Background: Children, being the future of any nation, not only need special attention but also need a suiTable environment to have proper growth and development. They are also vulnerable to various diseases and disabilities. It is observed that certain maternal characteristics are helpful in the development and survival of such high-risk children. Methodology: A community-based, cross-sectional, analytic study was undertaken among children under 5 years of age in two slums. The population of both slums was 1550, out of which children under 5 years of age were 196 (12.64%), which constituted the study population. The "high-risk" children were identified based on pre-development criteria of "risk factors." Data was collected by interview technique, physical examination of the children, and checking of medical documents. The data regarding "high-risk" children were analyzed to determine the association of "high-risk" children with maternal factors like birth order, education, and occupation of the mother and child being looked after as the mother's substitute. The data obtained was subjected to standard statistical methods to achieve valid comparisons. Results: The present study revealed that out of 196 children under 5 years of age surveyed, 88 were "high-risk" children which constituted a prevalence rate of 44.89%. A significant association had been found between the prevalence of "high-risk" children and factors like birth order, education, and occupation of the mother and child being looked after by the mother's substitute. Conclusion: The study showed a significant association between various maternal factors and high-risk children. Thus maternal social and environmental factors along with enhancing pregnancy health go a long way in preventing high-risk children and betterment of child health.