Rabbit muscle pyruvate kinase activators: Synthesis, molecular docking and theoretical studies of N-substituted sulfonamide derivatives.

Pyruvate kinase (PK) activators have potential therapeutic applications in diseases such as sickle cell anemia. In this study, N-Substituted sulfonamide derivatives of 1,4-dihydropyridines were synthesized and evaluated as PK activators in vitro and using molecular docking studies. The compounds were synthesized by reacting dicarbonyl compounds with ammonium acetate, 5-nitrobenzaldehyde, and alumina sulfuric acid (ASA), followed by reduction and sulfonylation. The structures of the compounds were analyzed using spectroscopic techniques. DFT calculations provided insights into the electronic properties. Molecular docking of the compounds into the active site of PK showed favorable binding interactions. ADME evaluation indicated suitable solubility, BBB permeation, and lack of CYP450 inhibition. Overall, this study demonstrates the potential of new hybrid 1,4-dihydropyridine substituted sulfonamides as PK activators for further development. According to AC50 values, the compound (DTSF7, 0.97µM) is about 100-fold higher affective than the clinically used sulfonamide compound (AC50 = 90µM) for PK.

Thymic gene expression analysis reveals a potential link between HIF-1A and Th17/Treg imbalance in thymoma associated myasthenia gravis.

Myasthenia gravis (MG) is an immune-mediated disease frequently associated with thymic changes. Increased T helper 17 (Th17) cell activity and dysfunctional regulatory T (Treg) cells have been demonstrated in subgroups of MG. On the other hand, hypoxia-inducible factor 1 (HIF-1) has been shown to regulate the Th17/Treg balance by inducing Th17 differentiation while attenuating Treg development. To identify the underlying mechanisms of different thymic pathologies in MG development, we evaluated thymic samples from thymoma-associated myasthenia gravis (TAMG), MG with hyperplasia (TFH-MG) and thymoma without MG (TOMA) patients. Differential gene expression analysis revealed that TAMG and TFH-MG cells are associated with different functional pathways. A higher RORC/FOXP3 ratio provided evidence for Th17/Treg imbalance in TAMG potentially related to increased HIF1A. The hypoxic microenvironment in thymoma may be a driver of TAMG by increasing HIF1A. These findings may lead to new therapeutic approaches targeting HIF1A in the development of TAMG.

Effect of Intrathecal Eugenol on Cerebral Vasospasm in an Experimental Subarachnoid Hemorrhage Model.

BACKGROUND: Eugenol has various curative properties. It affects the dilatation of cerebral arteries through voltage-dependent Ca2+ channel inhibition. This study is the first to explore the impact of eugenol on neuroprotection and vasospasm in an experimental subarachnoid hemorrhage (SAH) model. METHODS: Twenty-four adult male Sprague-Dawley rats were indiscriminately separated into 3 groups: the control group (n = 8), the SAH group (n = 8), and the eugenol group (n = 8). A double-bleeding method was used. The eugenol group received intracisternal eugenol (Sigma-Aldrich, St. Louis, MO, USA) at 30 µg/20 µl after induction of SAH. On the day 7, all groups were euthanized. Measurements were taken for basilar artery wall thickness, lumen diameter, serum endothelin-1 (ET-1), and caspase-3 levels. RESULTS: The eugenol group exhibited significantly lower wall thickness, ET-1, oxidative stress index, and caspase-3 levels compared to the SAH group. In comparison to the control group, the eugenol group showed a higher oxidative stress index along with higher ET-1 and caspase-3 levels, but these differences were not statistically significant. Wall thickness was significantly higher in the eugenol group than in the control group. CONCLUSIONS: This study represents the first literature exploration of intrathecal eugenol's impact on vasospasm induced after experimental SAH. Administration of intrathecal eugenol demonstrates a positive effect on the treatment of experimental vasospasm as well as on the reduction of oxidative stress and apoptosis.

Bovine carbonic anhydrase (bCA) inhibitors: Synthesis, molecular docking and theoretical studies of bisoxadiazole-substituted sulfonamide derivatives.

This paper describes the in vitro inhibition potential of bisoxadiazole-substituted sulfonamide derivatives (6a-t) against bovine carbonic anhydrase (bCA) after they were designed through computational analyses and evaluated the predicted interaction via molecular docking. First, in silico ADMET predictions and physicochemical property analysis of the compounds provided insights into solubility and permeability, then density functional theory (DFT) calculations were performed to analyse their ionization energies, nucleophilicity, in vitro electron affinity, dipole moments and molecular interactions under vacuum and dimethyl sulfoxide (DMSO) conditions. After calculating the theoretical inhibition constants, IC50 values determined from enzymatic inhibition were found between 12.93 and 45.77 µM. Molecular docking evaluation revealed favorable hydrogen bonding and π-interactions of the compounds within the bCA active site. The experimentally most active compound, 6p, exhibited the strongest inhibitory activity with a theoretical inhibition constant value of 9.41 nM and H-bonds with Gln91, Thr198, and Trp4 residues and His63 Pi-cation interactions with His63 residues. Overall, the study reveals promising bCA blocking potential for the synthesized derivatives, similar to acetazolamide.

Impact of Surgery-Induced Myeloid-derived Suppressor Cells and the NOX2/ROS Axis on Postoperative Survival in Human Pancreatic Cancer.

Preclinical studies imply that surgery triggers inflammation that may entail tumor outgrowth and metastasis. The potential impact of surgery-induced inflammation in human pancreatic cancer is insufficiently explored. This study included 17 patients with periampullary cancer [pancreatic ductal adenocarcinoma (PDAC) n = 14, ampullary carcinoma n = 2, cholangiocarcinoma n = 1] undergoing major pancreatic cancer surgery with curative intent. We analyzed the potential impact of preoperative and postoperative immune phenotypes and function on postoperative survival with >30 months follow-up. The surgery entailed prompt expansion of monocytic myeloid-derived suppressor cells (M-MDSC) that generated NOX2-derived reactive oxygen species (ROS). Strong induction of immunosuppressive M-MDSC after surgery predicted poor postoperative survival and coincided with reduced functionality of circulating natural killer (NK) cells. The negative impact of surgery-induced M-MDSC on survival remained significant in separate analysis of patients with PDAC. M-MDSC-like cells isolated from patients after surgery significantly suppressed NK cell function ex vivo, which was reversed by inhibition of NOX2-derived ROS. High NOX2 subunit expression within resected tumors from patients with PDAC correlated with poor survival whereas high expression of markers of cytotoxic cells associated with longer survival. The surgery-induced myeloid inflammation was recapitulated in vivo in a murine model of NK cell-dependent metastasis. Surgical stress thus induced systemic accumulation of M-MDSC-like cells and promoted metastasis of NK cell-sensitive tumor cells. Genetic or pharmacologic suppression of NOX2 reduced surgery-induced inflammation and distant metastasis in this model. We propose that NOX2-derived ROS generated by surgery-induced M-MDSC may be targeted for improved outcome after pancreatic cancer surgery. SIGNIFICANCE: Pancreatic cancer surgery triggered pronounced accumulation of NOX2+ myeloid-derived suppressor cells that inhibited NK cell function and negatively prognosticated postoperative patient survival. We propose the targeting of M-MDSC as a conceivable strategy to reduce postoperative immunosuppression in pancreatic cancer.

Isolated Transverse Process Fractures: Should We Offer Lumbar Corset or Not?

Background The aim of this study is to emphasize the need to be careful in terms of internal organ injuries in patients with isolated transverse process fracture (ITPF), and to investigate the effectiveness of corset use in controlling acute pain. Methods This is a retrospective study including 72 patients with only transverse process fractures secondary to trauma, who were admitted to the Emergency Department of Sakarya University Research and Training Hospital between January 2020 and October 2022. The radiological diagnoses were collected from spinal vertebral computed tomography images. Twelve patients were excluded from the study due to exclusion criteria. Sixty patients with ITPF were included in the comparison group. All patients were divided into two groups. The group with no lumbar corset (LC) included those who were discharged with analgesic and muscle relaxant treatment without a brace (n = 33). The LC+ group (n = 27) included those who received rigid lumbosacral orthosis in addition to analgesic and muscle relaxant treatment. Pain levels of all cases in both groups were evaluated with Visual Analog Scale scores on the day of trauma, the first week, the first month, and the sixth month. Results A total of 25 cases had one ITPF, 25 had two, 17 had three, and five patients had four or more ITPFs. The hospitalization rate was the highest among patients with four or more ITPFs (40%). Although the hospitalization rates according to the number of ITPFs were not statistically significant (p = 0.528), there was a clinical significance regarding increasing hospitalization rates with the increasing number of ITPFs. The hospitalization rates were 12%, 16%, 17.6%, and 40% in patients with one, two, three, and four or more ITPFs, respectively. Conclusion ITPFs should be treated conservatively. Concomitant organ injuries must be ruled out before treatment. Medical treatment without a lumbar corset could be used as a cost-effective choice.

Effect of intravenous lipid therapy in critically ill pediatric patients with calcium channel blocker toxicity.

BACKGROUND: Overdose with calcium-channel blockers (CCBs) still maintain their importance with a high lethality rate after exposure. We report the intravenous lipid emulsion therapy (ILE) therapy in our CCB overdose patients. METHODS: We retrospectively analyzed the records of 6 patients with CCB intoxication from Batman Training and Research Hospital PICU between March 2021 and September 2022. Patients aged 0-18 years who received ILE treatment for CCB poisoning were included. RESULTS: All six patients ingested CCB with the intention of committing suicide and were followed up in the pediatric intensive care unit (PICU). All patients received ILE therapy due to hemodynamic instability despite intravenous fluid boluses, calcium, glucagon, insulin-dextrose, and vasoactive agents. Vasoactive-Inotropic Score (VIS) decreased after ILE treatment. All patients were transferred from the PICU after recovery. CONCLUSIONS: ILE therapy should be kept in mind as a salvage therapy in hemodynamically unstable CCB poisoning cases that do not respond to initial and advanced options.

Bilateral Concha Bullosa Within Concha Bullosa: Unique Middle Concha Variation.

The middle nasal turbinate is an important anatomical formation located on the lateral nasal wall. Concha bullosa can be defined as the presence of an air gap inside the turbinate. It is the most common middle nasal turbinate variation. It is often asymptomatic, but can sometimes cause nasal obstruction. In this study, an asymptomatic patient with concha bullosa within concha bullosa in both middle turbinates, which has not been reported before in the literature, is presented. Laryngoscope, 134:3516-3518, 2024.

Unraveling the Impact of miRNA-17 in Glial Tumors and Cerebral Metastases: A Step Towards Enhanced Diagnosis and Prognosis.

BACKGROUND/AIM: MicroRNAs (miRNAs) have been identified as key regulators in various cancer types, including brain tumors. This study aimed to investigate the differential expression of miRNA-17 in glial tumors, cerebral metastases, and normal glial tissues. MATERIALS AND METHODS: A total of 42 patients were included in this cross-sectional study. Tissue samples were obtained from patients with glial tumors or cerebral metastases and from normal glial tissues. miRNA-17 expression levels were computed by using real-time polymerase chain reaction. Receiver operating characteristics analysis was used to determine the predictive potential of miRNA-17. RESULTS: In this study, we demonstrated a statistically significant difference in miRNA-17 expression levels between glial tumors and the control group (p=0.001), with higher miRNA-17 expression observed in glial tumors. Similarly, there was statistically higher miRNA-17 expression in metastatic cases compared with the control group (p=0.007). CONCLUSION: These findings suggest miRNA-17 might be a potential biomarker for differentiating glial tumors and cerebral metastases from normal glial tissue, although further research is necessary to validate these findings and investigate the potential role of miRNA-17 in the pathogenesis of these brain tumors.

Unconventional mobility, conventional trauma: a comparative analysis of e-scooter-related fractures.

PURPOSE: The present research aimed to compare the different types of fractures to those caused by e-scooter use as well as reveal the severity of e-scooter injuries via an epidemiological analysis of fractures over the course of 6 months. METHODS: This retrospective study assessed medical records of patients with fractures or dislocations admitted to a Turkish level three trauma centre emergency orthopaedic department between June 2021 and January 2022. Using a two-group design, the first group encompassed all fractures and dislocations treated, while the second focused on e-scooter-related cases. Comparative analysis covered fracture patterns and factors within the e-scooter group, like demographics, injury mechanism, and timing. Surgical need and types were explored for e-scooter injuries. RESULTS: Among 4481 upper extremity fractures, finger fractures (27.47%) and distal radius fractures (25.37%) were common, while e-scooter-related cases exhibited radius and ulna shaft fractures (23.07%). Of 2400 lower extremity fractures, toe fractures (30.2%) and metatarsal fractures (19.66%) predominated, with e-scooter-related injuries largely featuring metatarsal fractures (30%). The surgery rate in all patients was 8.92%, whereas the surgery rate specifically for e-scooter injuries was 48.2%. Significantly greater occurrence of lower extremity fractures was evident in e-scooter-related injuries compared to upper extremities (p = 0.011). Collisions involving stationary or moving objects were linked to injuries in the lower extremities, whereas falls were primarily associated with injuries in the upper extremities. Treatment included surgery (48.2%) and conservative management (52.8%), with ORIF (35.7%) and CRIF (10.7%) utilised. CONCLUSION: E-scooter-related fractures and dislocations are typically more severe, often requiring surgery, compared to other fractures during the same period. The study emphasises the link between injury type and increased risk of lower extremity fractures or dislocations.

Timing for extraction of permanent first molars in school aged children: a pilot study.

Extraction of permanent first molars (P1Ms) could create gaps, tipping and/or rotating of neighboring teeth, which depend on the extraction timing. This study evaluated the outcomes of P1Ms extraction in children who were classified based on their permanent second molar's (P2Ms) dental calcification at the time of extraction. In this retrospective study, the evaluations were made for 406 panoramic radiographs (PRs) of children aged 7-14 years who had single P1M extracted. Twenty-nine children having pre- and post-treatment PRs were selected based on the inclusion criteria. First, 2 groups were formed based on the extraction time by using the pre-treatment PRs; Early extraction group (EE) (n = 15) including Nolla Grades 5-6-7, and late extraction group (LE) (n = 14) including Nolla Grades 8-9-10 for P2Ms. Then, the inclination degrees of neighbouring teeth on extraction side and contralateral quadrant (non-extraction) side were measured by using the post-extraction PRs. Mann-Whitney U test was employed, and the statistical significance was set at p < 0.05. The mesial tipping tendency of P2M was significant towards the extraction side both at EE or LE calcification grades of P2Ms (p > 0.05). The mesial inclination degrees of maxillary P2Ms showed no statistically significant difference between the two sides of LE group. The ideal extraction time should be determined according to the dental age to plan an appropriate occlusal maintenance.

Fathering styles in a traditional culture and its association with marital relationship: A latent profile analysis with a nationally representative sample.

This study had two main objectives. The first goal was to examine fathers' parenting styles in a society with traditional patriarchal values. The second goal was to understand the extent to which marital relationship variables (i.e., marital satisfaction and spousal support) are related with the identified fathering profiles. The theoretical and empirical underpinnings of the "parenting styles" literature that form the foundation of the research on parent-child dynamics are still mainly based on data collected only from mothers. This weakness of the literature on fathering typologies is further accompanied by the scarcity of data on fathers' parenting practices from non-Western contexts. To achieve the study aims, a latent profile analysis based on warmth and control dimensions of parenting was performed using a nationally representative sample of 1070 urban fathers in Turkey. Using scores on six parenting behavior variables (i.e., warmth, punishment, inductive reasoning, positive parenting, discipline and teaching responsibilities, time and talking together), results revealed four fathering profiles: "authoritative" (30%), "average" (45%), "uninvolved" (16%), and "disciplining-distant" (8%). Furthermore, the results showed that the fathering profiles significantly differed from each other on the marital satisfaction and spousal support scores. Results highlight the importance of investigating fathering behaviors and their predictors in different cultures.

Evaluation of the relationship between pain inflammation due to dental caries and growth parameters in preschool children.

OBJECTIVES: To evaluate the relationship between pain inflammation due to dental caries and growth parameters, sleep disturbances, and oral health-related quality of life (OHRQoL) in preschool children before/after dental treatment and compare the results with the control group. MATERIALS AND METHODS: Study (pain inflammation due to caries) and control groups were included in this prospective clinical trial. The Child Sleep Habits Questionnaire (CSHQ) assessing sleep disturbances and the Early Childhood Oral Health Impact Scale (ECOHIS) assessing OHRQoL were applied in the corresponding time intervals to the study and control groups, respectively: baseline (T0study), 7 days after treatment (T1study), and following 6 months (T2study); baseline (T0control), and the following 6 months (T2control). Biochemical growth parameters (insulin-like growth factor-1 and insulin-like growth factor binding protein-3) and anthropometric measurements (standard deviation score of height, weight, and body mass index) were obtained at T0study, T2study, and T0control. Mann-Whitney U and the Student t-tests were used for statistical analyses. The significance level was set at p < 0.05. RESULTS: Data on 45 children (mean age: 55.6 ± 10.37 months) were analyzed. T2study was statistically higher than T0study for the anthropometric measurements and biochemical growth parameters (p < 0.05). T0study was statistically higher than T0control for biochemical growth parameters (p < 0.05). CSHQ and ECOHIS scores were found statistically significant at T0study than T0control (p < 0.05). Statistical scores of CSHQ and ECOHIS in T2study were significantly reduced compared to T0study (p < 0.05). CONCLUSION: Children's growth parameters, sleep disturbances, and OHRQoL improved after the elimination of pain and inflammation. CLINICAL RELEVANCE: This study's novelty is the observation of drastically increased growth parameters and reduced sleep disturbances following dental treatment.

D3SENet: A hybrid deep feature extraction network for Covid-19 classification using chest X-ray images.

Covid-19 is one of the biggest global epidemics seen in the world in recent years. Because of this, people's daily lifestyles, the economic conditions of countries and individuals, and most importantly, their health status has been adversely affected all over the world. Millions of people around the world have died from this disease. For this reason, rapid and accurate detection of the disease is of great importance in terms of treatment and precautions. In addition, it is especially important to correctly distinguish between Covid-19 and non-Covid-19 pneumonia diseases for correct diagnosis and treatment. These two diseases cause similar symptoms, and the symptoms and the effects of the disease on the body should be carefully examined for their differentiation. Chest X-ray images, chest computerized tomography, and swab tests are commonly used to detect patients infected with COVID-19. This disease affects the lungs the most in the body and causes fatal side effects such as shortness of breath. Therefore, medical images taken from the chest play an important role in the diagnosis of the disease. The fact that X-rays are faster and cheaper than computerized tomography has led to an increase in studies on the detection of disease with X-rays. In recent years, the impressive results of deep learning in the field of computer vision have attracted researchers to this field when working with image data. This study aims to detect these diseases on chest X-ray images collected from Covid-19 patients, pneumonia patients, and healthy individuals. We proposed a hybrid feature extraction network namely D3SENET which consists of DarkNet53, DarkNet19, DenseNet201, SqueezeNet, and EfficientNetb0. After a balanced data set was prepared, feature vectors were obtained from images using deep learning-based CNN models and the size of feature vectors was reduced by feature selection algorithms. Obtained features were classified by traditional machine learning methods such as SVMs. The number of features to be selected was tested by the iterative increment method and the parameters with the highest accuracy rate were obtained. As a result, it was seen that healthy and infected individuals were detected in 3 classes with an accuracy rate of 98.78%. In addition, the confusion matrix, precision, recall values, and F1 score of the obtained model are also given.

Design, Modeling and Fabrication of a Novel Class V Flextensional Transducer: The Sea-Shell.

In this study, a novel class V flextensional transducer (FT) was developed by assembling symmetrically convex ceramic and metal caps to form a seashell-like structure. The transducer was designed and analyzed using ATILA finite element analysis (FEA) software. The diameter, thickness, and radius of curvature of the ceramic and metal shells have been investigated as design parameters. The transducer was found to display four distinct flextensional modes in addition to the main radial resonance mode between 1 and 200 kHz frequency range. Prototypical devices were fabricated from four different commercial lead zirconate titanate (PZT) compositions and underwater performances were compared. Transmitting voltage responses (TVRs) were observed to range between 120 and 135 dB (ref 1 µ Pa/V) at frequencies above 50 kHz.

Nuclear factor erythroid-2-related factor 2 (Nrf2) is a potential prognostic factor in patients with gastric adenocarcinoma.

BACKGROUND: Gastric cancer is one of the leading causes of cancer-related death, and many researchers are focused on the discovery and use of different biomarkers in prognosis prediction. The use of Nrf2 as a prognostic marker in patients with gastric cancer remains controversial. In this study, we evaluated the expression of Nrf2 protein in gastric adenocarcinoma. PATIENTS AND METHODS: A total of 86 patients who underwent gastric resection and D2 lymph node dissection due to gastric adenocarcinoma were included. Clinicopathological characteristics, such as age, gender, gastrectomy type, pathologic T (pT) and N (pN) stages, tumor grade, tumor type per Lauren's classification, presence of lymphovascular invasion, and Nrf2 expression were evaluated. RESULTS: While pT, pN, and Nrf-2 expression were found to be negative prognostic predictors for overall survival in one-way analysis of variance, Nrf-2 expression was the only significant negative prognostic predictor in multivariance analysis. pT, pN, diffuse type, high tumor grade, and Nrf-2 expression significantly affected overall survival in Kaplan-Meier survival analyses (p = 0.02, p = 0.03, p < 0.01, p = 0.027, and p = 0.001, respectively). CONCLUSIONS: Our findings support that Nrf2 is a prognostic marker in patients with gastric adenocarcinoma. Anti-Nrf2 therapy options should be investigated to improve prognosis in gastric cancer patients.

The management of children aged 0-5 years with suspected foreign body aspiration: a prospective study.

INTRODUCTION: A simple algorithm for bronchoscopy was prepared in very young children. METHODS: The patients aged 0-5 years who were applied with bronchoscopy because of suspected foreign body aspiration (FBA) analysed. RESULTS: Evaluations were made of 89 patients, as 55 (61.7%) in the FBA (+) group and 34 (38.3%) in the FBA (-) group. FBA was determined most in the 1-2 years age group (28 patients, 50.9%, p = 0.04) due to organic hard foodstuffs (94.5%, p < 0.001). The parameters found to be significant were witnessing the event (OR 12.133, 95% CI 3.147-46.774, p < 0.001) and not obtaining unilateral respiratory sounds (OR 7.556, 95% CI 2.681-21.292, p < 0.001). The most significant diagnostic finding was the determination of unilateral hyperventilation on X-ray (OR 16.730, 95% CI 4.541-61.632, p < 0.001). The operating time and length of stay in hospital was significantly shorter in the FBA (-) patients (p < 0.001). CONCLUSION: The presence of a witness, not obtaining unilateral respiratory sounds, and unilateral hyperventilation seen on X-ray are indications for bronchoscopy. In FBA (-) patients applied with bronchoscopy, the complication rate associated with the procedure is low, and the operating time and length of stay in hospital are short.

Pseudo-Meigs syndrome secondary to endodermal sinus tumor.

Ovarian tumors are the most common gynecological tumors seen in girls. Approximately 60-70% of them are germ cell tumors. Pseudo-Meigs syndrome is characterized by the presence of pelvic tumoral mass (benign or malign), pleural effusion, and massive acid. If the tumor is removed, acid and hydrothorax disappear. Endodermal sinus (yolk sac) tumor is a very rare cause in the diagnosis of Pseudo-Meigs syndrome, and only a few cases have been reported. This case is one of the rare cases presenting with Pseudo-Meigs syndrome and pathologically diagnosed as yolk sac tumor.

Exosome Structures Supported by Machine Learning Can Be Used as a Promising Diagnostic Tool.

Principal component analysis (PCA) as a machine-learning technique could serve in disease diagnosis and prognosis by evaluating the dynamic morphological features of exosomes via Cryo-TEM-imaging. This hypothesis was investigated after the crude isolation of similarly featured exosomes derived from the extracellular vehicles (EVs) of immature dendritic cells (IDCs) JAWSII. It is possible to identify functional molecular groups by FTIR, but the unique physical and morphological characteristics of exosomes can only be revealed by specialized imaging techniques such as cryo-TEM. On the other hand, PCA has the ability to examine the morphological features of each of these IDC-derived exosomes by considering software parameters such as various membrane projections and differences in Gaussians, Hessian, hue, and class to assess the 3D orientation, shape, size, and brightness of the isolated IDC-derived exosome structures. In addition, Brownian motions from nanoparticle tracking analysis of EV IDC-derived exosomes were also compared with EV IDC-derived exosome images collected by scanning electron microscopy and confocal microscopy. Sodium-Dodecyl-Sulphate-Polyacrylamide-Gel-Electrophoresis (SDS-PAGE) was performed to separate the protein content of the crude isolates showing that no considerable protein contamination occurred during the crude isolation technique of IDC-derived-exosomes. This is an important finding because no additional purification of these exosomes is required, making PCA analysis both valuable and novel.