Identification of dysregulation of sphingolipids in retinoblastoma using liquid chromatography-mass spectrometry.

Retinoblastoma (RB) is a rare ocular cancer seen in children that counts for approximately 3% of all childhood cancers. It is found that mutation in RB1, a tumour Suppressor Gene on chromosome 13 as the cause of malignancy. Retinoblastoma protein is the target for ceramide to cause apoptosis. We studied lipidomics of two RB cell lines, one aggressive cell line (NCC-RbC-51) derived from a metastatic site and one non aggressive cell line (WERI-Rb1) in comparison with a control cell line (MIO-M1). Lipid profiles of all the cell lines were studied using high resolution mass spectrometer coupled to high performance liquid chromatography. Data acquired from all the three cell lines in positive mode were analyzed to identify differentially expressed metabolites. Several phospholipids and lysophospholipids were found to be dysregulated. We observed upregulation of hexosyl ceramides, and down regulation of dihydroceramides and higher order sphingoglycolipids hinting at a hindered sphingolipid biosynthesis. The results obtained from liquid chromatography-mass spectrometry are validated by using qPCR and it was observed that genes involved in ceramide biosynthesis pathway are getting down regulated.

Synthesis and Characterization of a Novel Peptide Targeting Human Tenon Fibroblast Cells To Modulate Fibrosis: An Integrated Empirical Approach.

Fibrosis is the primary factor influencing the prognosis of glaucoma post-trabeculectomy surgery, an eye condition characterized by increased intraocular pressure (IOP). Despite advancements in surgical procedures and aftercare, it continues to be a serious impediment. During the clinical intervention of scarring, fibrosis is managed by using topical application of combined antifibrotic drugs (mitomycin C). But still, scarring remains a key problem due to minimal drug penetration and nonbioavailability. In this study, we synthesized a cell-specific peptide for modulating scarring in human tenon fibroblasts undergoing epithelial-mesenchymal transition (EMT). The peptide was also conjugated with mitomycin C in order to investigate the effect of the drug conjugation on human tenon fibroblasts from the nanofiber composite system and to evaluate the fibrosis process. Peptide VRF2019 was identified using a subtractive proteomics approach, including solubility, cell penetration, and amphipathic properties. The peptide structure was determined using circular dichroism spectroscopy. The peptide and drug was conjugated using N-ethyl-N'-(3-(dimethylamino)propyl) carbodiimide/N-hydroxysuccinimide (EDC-NHS) chemistry, and the conjugation efficiency was evaluated using high-pressure liquid chromatography. The conjugated product and polycaprolactone (PCL) were electrospun to form a composite nanofiber, which was tested for cytotoxicity and drug release on human tenon fibroblast cells. The modeled VRF2019 peptide structure formed an α-helical structure with all residues spanning the allowed regions of the Ramachandran plot. Subsequent molecular dynamics simulations also demonstrated its membrane penetration potential. The peptide uptake was also studied in human tenon fibroblast cells. High-pressure liquid chromatography (HPLC) and mass spectrometry measurements confirmed peptide-drug conjugation and stability. Furthermore, scanning electron microscopy (SEM) investigation revealed the structure and size of the PCL composite nanofiber. We infer from early research that the PCL composite nanofiber matrix can greatly increase drug delivery and bioavailability.

Elucidating the Therapeutic Potential of Cell-Penetrating Peptides in Human Tenon Fibroblast Cells.

Cell-penetrating peptides (CPPs) have been widely used as vehicles for delivering therapeutic molecules to the site of action. Apart from their delivering potential, the biological effects of CPPs have not been explored in detail. JTS-1 is a CPP that has been reported to have gene delivery functions, although its biological role is yet to be determined. Hence, in this study, we revealed the biological mechanism such as its uptake mechanism and immunogenic potential and function using primary human tenon fibroblast (TF) cells collected from patients undergoing glaucoma trabeculectomy surgery. Our results showed that the JTS-1 peptide has an α-helical structure and is nontoxic up to 1 µM concentration. It was found to be colocalized with early endosome (Rab5), recycling endosome (Rab7), and Rab11 and interacted with major histocompatibility complex (MHC) class I and II. The peptide also affected actin polymerization, which is regulated by cofilin phosphorylation and ROCK1 localization. It also inhibited TF cell proliferation. Therefore, the JTS-1 peptide could be used as a possible therapeutic agent for modifying the fibrosis process, where TF proliferation is a key cause of surgery failure.

Understanding the cellular response of human tenon fibroblast on polycaprolactone-Aloe vera blend fiber.

The signalling response is determined by the cell's reaction to different biochemical and biophysical inputs such as stiffness, topological, and structural alignment. The surface patterns at the nano-scale can be an influential factor in cell signalling behaviour. It is important to understand the cellular response to the biophysical cues for biomedical applications. Biomaterials have an important role in regenerative tissue engineering. In this study, we have fabricated electrospun polycaprolactone (PCL) and PCL-Aloe vera (PCL-AV) nanofibrous matrix and studied its effect on the human tenon fibroblast (HTF) cellular and morphological changes. The electrospun fibers were characterized using Scanning Electron Microscope (SEM), Fourier Transform Infrared spectroscopy (FTIR), Atomic Force Microscopy (AFM) and Brunaur, Emette and Teller (BET) analysis for their morphology, composition, topography, surface area and porosity. The results revealed fiber size, roughness and porosity has been altered by addition of AV. The HTF cell viability, proliferation and expression of focal adhesion proteins, such as FAK, Ezrin, Vasp and Cofilin on the PCL-AV fiber matrix were examined. The results showed a change in cellular morphology and a significant change in the cofilin phosphorylation on PCL-AV nanofiber. The influence of Aloe vera composition on the nano-dimension of the PCL has made a significant impact on the cellular morphology at both gene and protein levels. This observation suggests that AV composition in the nanofiber can significantly influence the HTF cellular adhesions.

Clinical Significance of Liver Function Abnormality in Patients with COVID-19: A Single-center Experience from Western India.

BACKGROUND AND AIMS: The impact of coronavirus disease-2019 (COVID-19) on liver function remains to be fully elucidated. This study was designed to investigate such and determine the clinical significance in determining mortality risk. METHODS: A retrospective study was conducted in patients with COVID-19 from March 2020 to July 2020. Clinical details were retrieved from electronic medical records to obtain clinical characteristics, medical history, laboratory tests, therapeutic intervention, and outcome data. RESULTS: A total of 184 patients with COVID-19 were included (median age: 45.5 years), comprised of 62.5% men. In total, 22 (12.0%) patients had severe infection and 162 (88.0%) had mild to moderate infection. Overall, 95 (51.6%) showed abnormal liver function test (LFT) and 17 (9.2%) showed normal LFT at admission. The median age, hospital stay, and LFT were significantly higher in severe vs. non-severe infection (p<0.001). Out of 12 deaths, the majority were due to severe infection (n=11). Deaths were also due to acute respiratory distress syndrome (n=5), cardiac reasons (n=3), and sepsis with multiorgan failure (n=3). The median age, hospital stay and number of intensive care unit admissions were higher in patients having abnormal LFT compared to normal LFT. Incidence of elevated aspartate aminotransferase (42.8% and 40.4%), alanine transaminase (43.7% and 41.6%), and hypoalbuminemia (71.4% and72.7%) at admission and discharge were more common in severe infection. The mean survival was significantly lower in severe infection compared to those with non-severe disease (17.2 vs. 52.3 days; p<0.001). CONCLUSIONS: Incidence of abnormal liver function was higher in patients with severe COVID-19 and was associated with prolonged hospital stay; mortality was associated with severity of COVID-19. For ruling out the risk of liver injury, it is crucial to vigilantly monitor the liver function parameters in patients with COVID-19 admitted to hospital.

A Rare Case of Left Atrial Myxoma Vascularity Causing Acquired Coronary Cameral Fistula: Role of Transesophageal Echocardiography.

Left atrial myxoma is the common benign tumor of heart. Coronary arteries may supply these tumor. Considering the vascular nature of the tumor, acquired coronary cameral fistula(CCF) can be a possibility postexcision of the left atrial myxoma. Here, we discuss a case of 53-year-old female patient, who developed acquired CCF, following excision of the tumor and the role of transesophageal echocardiography intraoperatively in diagnosis.

Non-Catheter Related Perioperative Superior Vena Cava Syndrome Following Surgical VSD Closure-Role of Transesophageal Echocardiography.

Unintentional iatrogenic surgical complications can complicate the operative and postoperative courses of paediatric cardiac surgery patients. Unless recognized and treated early, it's possible, these complications may lead to a prolonged hospital stay, increased treatment cost, morbidity, and even death. Ventricular Septal Defect (VSD) is the most common congenital heart defect in children, occurring in 50% of all children with congenital heart disease and in 20% as an isolated lesion. Herein, we discuss the development of Superior Vena Cava (SVC) syndrome following surgical repair of VSD in a nine-month-old child. Early clinical diagnosis, immediate confirmation with Transthoracic Echocardiography (TTE) and surgical re-exploration led to a good outcome in this patient. If, however, we could have performed an intraoperative Trans Esophageal Echocardiography (TEE), this complication could have been recognized, even before shifting from the operating room.

Feasibility of measuring superior mesenteric artery blood flow during cardiac surgery under hypothermic cardiopulmonary bypass using transesophageal echocardiography: An observational study.

BACKGROUND: Abdominal complications being rare but results in high mortality, commonly due to splanchnic organ hypoperfusion during the perioperative period of cardiac surgery. There are no feasible methods to monitor intraoperative superior mesenteric artery blood flow (SMABF). Hence, the aim of this study was to evaluate the feasibility and to measure SMABF using transesophageal echocardiography (TEE) during cardiac surgery under hypothermic cardiopulmonary bypass (CPB). METHODOLOGY: Thirty-five patients undergoing elective cardiac surgery under CPB were enrolled. Heart rate, mean arterial pressure (MAP), cardiac output (CO), SMABF, superior mesenteric artery (SMA) diameter, superior mesentric artery blood flow over cardiac output (SMA/CO) ratio and arterial blood lactates were recorded at three time intervals. T0: before sternotomy, T1: 30 min after initiation of CPB and T2: after sternal closure. RESULTS: SMA was demonstrated in 32 patients. SMABF, SMA diameter, SMA/CO, MAP and CO-decreased significantly (P < 0.0001) between T0 and T1, increased significantly ( P ≤ 0.001) between T1 and T2 and no significant change (P > 0.05) between T0 and T2. Lactates increased progressively from T0 to T2. CONCLUSION: Study shows that there is decrease in SMABF during CPB and returns to baseline after CPB. Hence, it is feasible to measure SMABF using TEE in patients undergoing cardiac surgery under hypothermic CPB. TEE can be a promising tool in detecting and preventing splanchnic hypoperfusion during perioperative period.

Evaluation of adjusted central venous blood gases versus arterial blood gases of patients in post-operative paediatric cardiac surgical intensive care unit.

BACKGROUND AND AIMS: Central venous catheters are in situ in most of the intensive care unit (ICU) patients, which may be an alternative for determining acid-base status and can reduce complications from prolonged arterial cannulation. The aim of this study was to examine the reliability between adjusted central venous blood gas (aVBG) and arterial blood gas (ABG) samples for pH, partial pressure of carbon-di-oxide (pCO2), bicarbonate (HCO3 (-)), base excess (BE) and lactates in paediatric cardiac surgical ICU. METHODS: We applied blood gas adjustment rule, that is aVBG pH = venous blood gas (VBG) pH +0.05, aVBG CO2 = VBG pCO2 - 5 mm Hg from the prior studies. In this study, we validated this relationship with simultaneous arterial and central venous blood obtained from 30 patients with four blood sample pairs each in paediatric cardiac surgical ICU patients. RESULTS: There was a strong correlation (R i.e., Pearson's correlation) between ABG and aVBG for pH = 0.9544, pCO2 = 0.8738, lactate = 0.9741, HCO3 (-) = 0.9650 and BE = 0.9778. Intraclass correlation co-efficients (ICCs) for agreement improved after applying the adjustment rule to venous pH (0.7505 to 0.9454) and pCO2 (0.4354 to 0.741). Bland Altman showed bias (and limits of agreement) for pH: 0.008 (-0.04 to + 0.057), pCO2: -3.52 (-9.68 to +2.65), lactate: -0.10 (-0.51 to +0.30), HCO3 (-): -2.3 (-5.11 to +0.50) and BE: -0.80 (-3.09 to +1.49). CONCLUSION: ABG and aVBG samples showed strong correlation, acceptable mean differences and improved agreement (high ICC) after adjusting the VBG. Hence, it can be promising to use trend values of VBG instead of ABG in conjunction with a correction factor under stable haemodynamic conditions.

Vegetation in Bangalore's Slums: Composition, Species Distribution, Density, Diversity, and History.

There is widespread acknowledgement of the need for biodiversity and greening to be part of urban sustainability efforts. Yet we know little about greenery in the context of urban poverty, particularly in slums, which constitute a significant challenge for inclusive development in many rapidly growing cities. We assessed the composition, density, diversity, and species distribution of vegetation in 44 slums of Bangalore, India, comparing these to published studies on vegetation diversity in other land-use categories. Most trees were native to the region, as compared to other land-use categories such as parks and streets which are dominated by introduced species. Of the most frequently encountered tree species, Moringa oleifera and Cocos nucifera are important for food, while Ficus religiosa plays a critical cultural and religious role. Tree density and diversity were much lower in slums compared to richer residential neighborhoods. There are also differences in species preferences, with most plant (herb, shrub and vines) species in slums having economic, food, medicinal, or cultural use, while the species planted in richer residential areas are largely ornamental. Historic development has had an impact on species distribution, with older slums having larger sized tree species, while recent slums were dominated by smaller sized tree species with greater economic and food use. Extensive focus on planting trees and plant species with utility value is required in these congested neighborhoods, to provide livelihood support.

Comparison of left internal mammary artery diameter before and after left stellate ganglion block.

AIMS AND OBJECTIVES: Left internal mammary artery (LIMA) is the preferred arterial conduit for coronary artery bypass grafting. Various pharmacological agents are known to increase LIMA blood flow. Sympathetic blockade mediated by stellate ganglion block (SGB) has been used to provide vasodilatation in the upper extremities and in the treatment of refractory angina. We investigated effect of left stellate ganglion block (LSGB) on LIMA diameter. MATERIALS AND METHODS: In 30 diagnosed patients of triple vessel coronary artery disease, LSGB was given under fluoroscopic guidance by C6 transverse process approach using 10 ml of 1% lignocaine. LIMA diameter was measured before and 20 min after the block at 2 nd , 3 rd , 4 th and at 5 th rib level. Heart rate (HR), systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean arterial pressure (MAP) were recorded before and 20 min after the block. RESULTS: The LIMA diameter increased significantly at 2 nd (2.56 ± 0.39 vs. 2.99 ± 0.40; P < 0.0001), 3 rd (2.46 ± 0.38 vs. 2.90 ± 0.40; P < 0.0001), 4 th (2.39 ± 0.38 vs. 2.84 ± 0.41; P < 0.0001) and 5 th rib level (2.35 ± 0.38 vs. 2.78 ± 0.40; P < 0.0001). No statistically significant change occurred in HR, SBP, DBP and MAP before and 20 min after LSGB. CONCLUSIONS: LSGB significantly increased the LIMA diameter. The LSGB can be considered as an alternative to topical and systemic vasodilators for reducing vasospasm of LIMA.