Significant Tissue Injury Research Articles

ISMRM Clinical Focus Meeting 2023: "Imaging the Fire in the Brain".

Set during the Annual Meeting of the International Society for Magnetic Resonance in Medicine (ISMRM), the "Clinical Focus Meeting" (CFM) aims to bridge the gap between innovative magnetic resonance imaging (MRI) scientific research and daily patient care. This initiative is dedicated to maximizing the impact of MRI technology on healthcare outcomes for patients. At the 2023 Annual Meeting, clinicians and scientists from across the globe were invited to discuss neuroinflammation from various angles (entitled "Imaging the Fire in the Brain"). Topics ranged from fundamental mechanisms and biomarkers of neuroinflammation to the role of different contrast mechanisms, including both proton and non-proton techniques, in brain tumors, autoimmune disorders, and pediatric neuroinflammatory diseases. Discussions also delved into how systemic inflammation can trigger neuroinflammation and the role of the gut-brain axis in causing brain inflammation. Neuroinflammation arises from various external and internal factors and serves as a vital mechanism to mitigate tissue damage and provide neuroprotection. Nonetheless, excessive neuroinflammatory responses can lead to significant tissue injury and subsequent neurological impairments. Prolonged neuroinflammation can result in cellular apoptosis and neurodegeneration, posing severe consequences. MRI can be used to visualize these consequences, by detecting blood-brain barrier damage, characterizing brain lesions, quantifying edema, and identifying specific metabolites. It also facilitates monitoring of chronic changes in both the brain and spinal cord over time, potentially leading to better patient outcomes. This paper represents a summary of the 2023 CFM, and is intended to guide the enthusiastic MR user to several key and novel sequences that MRI offers to image pathophysiologic processes underlying acute and chronic neuroinflammation. EVIDENCE LEVEL: 5 TECHNICAL EFFICACY: Stage 3.

Does cardiac impairment develop in ischemic renal surgery in rats depending on the reperfusion time?

BackgroundRenal dysfunction is known to cause heart failure. However, renal dysfunction associated with kidney surgeries (mediated by reperfusion injury) that affects the cardiac physiological function, especially during the recovery and repair phase of renal surgery is unknown. MethodMale Wistar rats (238 ± 18 g) were subjected to renal sham and ischemia-reperfusion (IR-bilateral clamping for 15 min/45 min and reperfusion for 24 h/48 h/7 days) surgeries. At the end of the experiment, the heart was isolated from the animal (to exclude neurohormonal influence) and perfused for 60 min with Krebs-Hanseleit buffer to study the physiological changes. ResultRenal artery bilateral occlusion for 45 min that creates ischemia, followed by 24 h of reperfusion did not impart any significant cardiac physiological functional decline but 48 h of reperfusion exhibited a significant decline in cardiac hemodynamic indices (Rate pressure product in x104 mmHg*beats/min: Sham- 3.53 ± 0.19, I45_R48–2.82 ± 0.21) with mild tissue injury. However, 7 days of reperfusion inflict significant physiological decline (Rate pressure product in x104 mmHg*beats/min - 2.5 ± 0.14) and tissue injury (Injury score- 4 ± 1.5) in isolated rat hearts. Interestingly, when the renal artery bilateral occlusion time was reduced to 15 min the changes in the hearts were negligible after 7 days. Cellular level exploration reveals a positive relation between functional deterioration of mitochondria and elevated mitochondrial oxidative stress and inflammation with cardiac physiological decline and injury linked with renal ischemia-reperfusion surgery. ConclusionCardiac functional decline associated with renal surgery is manifested during renal repair or recovery. This decline depends on cardiac mitochondrial health, which is negatively influenced by the renal IR mediators and kidney function.

Cardiac damage following renal ischemia reperfusion injury increased with excessive consumption of high fat diet but enhanced the cardiac resistance to reperfusion stress in rat

Renal ischemia-reperfusion (IR) injury inflicts remote cardiac dysfunction. Studies on rats fed with a high-fat diet (HD) showed contradictory results: some demonstrated increased sensitivity of the heart and kidney to IR injury, while others reported resistance. In this study, we examined cardiac dysfunction and compromised cardiac tolerance associated with renal IR in HD and standard diet (SD) fed rats. Male Wistar rats fed with HD or SD diet for 16 weeks were subjected to either renal sham or IR protocol (bilateral clamping for 45 min and reperfusion for 24 h). The hearts isolated from these rats were further subjected to normal perfusion or IR procedure to study cardiac response. Renal IR surgery negatively affected cardiac function with substantial changes in the cardiac tissues, like mitochondrial dysfunction, elevated oxidative stress, and inflammation. HD-fed rat hearts exhibited hypertrophy at the end of 16 weeks, and the consequential impact on the heart was higher in the animals underwent renal IR surgery than with sham surgery. However, the IR induction in the isolated heart from renal sham or renal IR operation showed significant tissue injury resistance and better physiological recovery in HD-fed rats. However, in SD-fed rats, only hearts from renal IR-operated rats showed resistance to cardiac IR, whereas hearts from renal sham-operated rats were more susceptible to IR damage. The augmented IR resistance in the heart with prior renal surgery was due to preserved mitochondrial bioenergetics function, reduced oxidative stress, and activation of the PI3K/AKT signaling axis.

Peripheral Administration of Norepinephrine: A Prospective Observational Study

Historically, norepinephrine has been administered through a central venous catheter (CVC) because of concerns about the risk of ischemic tissue injury if extravasation from a peripheral IV catheter (PIVC) occurs. Recently, several reports have suggested that peripheral administration of norepinephrine may be safe. Can a protocol for peripheral norepinephrine administration safely reduce the number of days a CVC is in use and frequency of CVC placement? This was a prospective observational cohort study conducted in the medical ICU at a quaternary care academic medical center. A protocol for peripheral norepinephrine administration was developed and implemented in the medical ICU at the study site. The protocol was recommended for use in patients who met prespecified criteria, but was used at the treating clinician's discretion. All adult patients admitted to the medical ICU receiving norepinephrine through a PIVC from February 2019 through June 2021 were included. The primary outcome was the number of days of CVC use that were avoided per patient, and the secondary safety outcomes included the incidence of extravasation events. Six hundred thirty-five patients received peripherally administered norepinephrine. The median number of CVC days avoided per patient was 1 (interquartile range, 0-2days per patient). Of the 603 patients who received norepinephrine peripherally as the first norepinephrine exposure, 311 patients (51.6%) never required CVC insertion. Extravasation of norepinephrine occurred in 35 patients (75.8 events/1,000d of PIVC infusion [95%CI, 52.8-105.4 events/1,000d of PIVC infusion]). Most extravasations caused no or minimal tissue injury. No patient required surgical intervention. This study suggests that implementing a protocol for peripheral administration of norepinephrine safely can avoid 1 CVC day in the average patient, with 51.6%of patients not requiring CVC insertion. No patient experienced significant ischemic tissue injury with the protocol used. These data support performance of a randomized, prospective, multicenter study to characterize the net benefits of peripheral norepinephrine administration compared with norepinephrine administration through a CVC.

A phase 2, open-label, adaptive, dose-ranging study with long-term extension to evaluate the safety, tolerability, efficacy, and pharmacokinetics of intra-articular AMB-05X injections in patients with tenosynovial giant cell tumor.

TPS11585 Background: CSF1R is a clinically validated target for Tenosynovial Giant Cell Tumor (TGCT). CSF1 is overexpressed in TGCT and, via action at the colony-stimulating factor 1 receptor (CSF1R), leads to the proliferation of synovial-like mononuclear cells and recruitment of multinucleate giant cells, foam cells, siderophages, and inflammatory cells, which compose the bulk of the neoplasm. Although nonmetastatic in nature, TGCT can develop into locally destructive lesions that cause significant local tissue injury, loss of joint function, and impaired quality of life. When complete resection is achievable without significant morbidity, the standard of care for patients with TGCT is surgical resection. Recurrence after surgery occurs in up to 30% of patients with localized TGCT and 83% of patients with diffuse TGCT ( Mastboom 2019, Patel 2017). Repeat surgeries may be necessary but often leads to further morbidity, complications, and reduced function of affected joints. A safe, effective, and convenient pharmacotherapy is a desirable addition to the therapeutic armamentarium for TGCT. AMB-05X is a potent and selective anti-CSF1R monoclonal antibody in development for local treatment of TGCT. Intra-articular (IA) injection of AMB-05X enables high drug concentration at the tumor site while minimizing systemic exposure and toxicity. A proof-of-concept study of IA AMB-05X for TGCT found clinically meaningful improvement in objective response rate, functional (pain, stiffness, range of motion) and quality of life measures after 12 weeks of therapy with no serious and a low number of adverse events associated with targeting CSF1R. This phase 2 study aims to establish the optimal dosing regimen and local administration of AMB-05X and confirm the efficacy and safety of 24 weeks and longer treatment. Methods: AMB-051-07 (NCT05349643) is an open-label, multicenter, phase 2 study of AMB-05X in patients with TGCT. The trial is being conducted at multiple sites in North America, Europe, and Australia. The dosing schedule includes an injection interval of every 4 weeks and longer, including an extended dosing period. Adult patients with histologically confirmed symptomatic TGCT with one joint involvement whether surgically amenable or non-amenable with measurable disease and adequate organ function are being enrolled. After the initial cohort of patients with TGCT of the knee, other joints may be enrolled pending evaluation by the DMC. The primary study outcome measures are objective response rate (ORR) per RECIST v1.1 by central radiology review and safety. Secondary outcome measures include ORR per modified RECIST and Tumor Volume Score, duration of response, time to response, PRO/QOL (including range of motion, stiffness, pain), pharmacodynamics and pharmacokinetics. Clinical trial information: NCT05349643 .

Serum miRNA profiles are altered in patients with primary sclerosing cholangitis receiving high-dose ursodeoxycholic acid

Background & AimsPrimary sclerosing cholangitis (PSC) is a chronic, progressive cholestatic liver disease that can lead to end-stage liver disease and cholangiocarcinoma. High-dose ursodeoxycholic acid (hd-UDCA, 28–30 mg/kg/day) was evaluated in a previous multicentre, randomised placebo-controlled trial; however, the study was discontinued early because of increased liver-related serious adverse events (SAEs), despite improvement in serum liver biochemical tests. We investigated longitudinal changes in serum miRNA and cytokine profiles over time among patients treated with either hd-UDCA or placebo in this trial as potential biomarkers for PSC and response to hd-UDCA, as well as to understand the toxicity associated with hd-UDCA treatment. MethodsThirty-eight patients with PSC were enrolled in a multicentred, randomised, double-blinded trial of hd-UDCA vs. placebo. ResultsSignificant alterations in serum miRNA profiles were found over time in both patients treated with hd-UDCA or placebo. Additionally, there were striking differences between miRNA profiles in patients treated with hd-UDCA compared with placebo. In patients treated with placebo, the changes in concentration of serum miRNAs miR-26a, miR-199b-5p, miR-373, and miR-663 suggest alterations of inflammatory and cell proliferative processes consistent with disease progression. However, patients treated with hd-UDCA exhibited a more pronounced differential expression of serum miRNAs, suggesting that hd-UDCA induces significant cellular miRNA changes and tissue injury. Pathway enrichment analysis for UDCA-associated miRNAs suggested unique dysregulation of cell cycle and inflammatory response pathways. ConclusionsPatients with PSC have distinct miRNAs in the serum and bile, although the implications of these unique patterns have not been studied longitudinally or in relation to adverse events related to hd-UDCA. Our study demonstrates marked changes in miRNA serum profiles with hd-UDCA treatment and suggests mechanisms for the increased liver toxicity with therapy. Impact and implicationsUsing serum samples from patients with PSC enrolled in a clinical trial comparing hd-UDCA with placebo, our study found distinct miRNA changes in patients with PSC who are treated with hd-UDCA over a period of time. Our study also noted distinct miRNA patterns in patients who developed SAEs during the study period.

Captive Black Mamba (Dendroaspis Polylepis) Bite Leading to Respiratory Failure

BackgroundOf the 8000–10,000 snake envenomations evaluated in U.S. emergency departments (ED) annually, approximately 1% are due to non-native snakes. We describe a 26-year-old man who was bitten by his captive black mamba (Dendroaspis polylepis) as he was packing it up for transport to another snake collector. Case ReportThe patient presented to the ED 1 h after being bitten on the forearm, complaining of left arm pain, oral paresthesias, and dyspnea. His vital signs: heart rate 96 beats/min, blood pressure 167/101 mm Hg, temperature 36.7°C (97.9°F), respiratory rate 20 breaths/min, and room air oxygen saturation 100%. Two mildly tender puncture wounds without swelling or ecchymosis were found on the posterior aspect of the forearm. Over the ensuing 30 min his dyspnea worsened, and he developed objective weakness. He was intubated and placed on mechanical ventilation. He was treated with atropine 2 mg for bronchorrhea. Five vials of South African Vaccine Producers (Johannesburg, South Africa) polyvalent antivenom were administered 2.5 h post-bite and the patient was admitted to the intensive care unit. He was extubated 18 h post-envenomation and discharged the following day. He has remained asymptomatic since leaving the hospital. Why Should an Emergency Physician Be Aware of This?The primary manifestations of D. polylepis envenomings are neurological. Initial signs may include paresthesias, dysarthria, dysphagia, and ptosis. Progressive descending paralysis leading to respiratory failure develops within 60 min. Muscarinic features are frequently observed. Cardiotoxicity and hematologic laboratory abnormalities may be present. Although pain is common, significant local tissue injury does not occur. In addition to supportive care, several non-native antivenoms are indicated for D. polylepis envenomations. Black mamba envenomings differ from the native snakebites with which U.S. physicians are familiar. Rapid, progressive neurological toxicity and muscarinic features are most common. Treatment consists of supportive care and appropriate antivenom administration.

Cardioprotective Effects of Octreotide against Sepsis-Induced Cardiotoxicity in Mice.

Sepsis is a systemic inflammatory consequence resulting from microbial infection, assessed as a worldwide healthcare issue. Sepsis can result in multiorgan dysfunction, including cardiac, renal, hepatic, and cerebral dysfunction. Cardiotoxicity can occur in humans and rodents during sepsis, leading to increased mortality. The current study aims to explore the possible cardioprotective effects of octreotide during sepsis-induced cardiotoxicity. This study was done with a total of forty male albino Swiss mice, aged 8-12 weeks and weighing 25-30 gm. These animals had free access to food and water. After two weeks of adaptation, mice were divided into four groups (n=10): 1) Normal group: healthy mice; 2) CLP group: mice underwent CLP operation; 3) Vehicle group: mice received DMSO. 4) Octreotide group: mice received octreotide (10 mg/kg) subcutaneously in 2 divided doses for 5 consecutive days. All groups underwent CLP operation on the 4th day, then sacrificed on the 5th day then blood, and tissue sampling was done. The Octreotide group demonstrated a significant (P<0.05) decrease in the myocardial levels of cardiac troponin-I as compared to the CLP group. Furthermore, the octreotide group demonstrated a significant (P<0.05) decrease in the serum level of inflammatory cytokines (TNF-α, IL-6, & IL-1β) as compared to the CLP group. Additionally, the octreotide group showed a significant (P<0.05) elevation in the myocardial activity of SOD and a reduction in MDA level compared to the CLP group. Histologically, all mice in the CLP group showed a significant (P<0.05) cardiac tissue injury, while the octreotide groups showed a significant (P<0.05) reduced level of cardiac tissue injury. The results of the present study revealed that octreotide attenuates sepsis-induced cardiotoxicity through different protective effects; they include the anti-inflammatory effect through their ability to decrease serum levels of inflammatory cytokines (TNF-α, IL-1β, and IL-6). Also, the anti-oxidant effect through their ability to decrease myocardial levels of MDA and increase the myocardial activity of SOD. Additionally, the direct cardiac protective effect through the lower level of cardiac troponin- I and the reduction of histopathological changes during sepsis-induced cardiotoxicity.

Protective effects of hesperidin in cyclophosphamide-induced parotid toxicity in rats

Cyclophosphamide (CYP) is an alkylating agent that is used on a wide range as a treatment of malignancies and autoimmune diseases. Previous studies have shown the promising role of hesperidin (HSP) as an antioxidant agent against various models of toxic agents. The protective effect of the HSP against CYP-induced parotid damage was evaluated in this study. Forty rats (180–200 g) were divided into four equal groups: Group I (received normal saline), Group II (HSP-treated at a dose of 100 mg/kg/day for 7 consecutive days), Group III (CYP-treated at a dose of 200 mg/kg single intraperitoneal injection on the 7th day of the experiment), Group IV (CYP + HSP); HSP-treated at a dose of 100 mg/kg/day for 7 consecutive days and CYP (200 mg/kg) single intraperitoneal injection on the 7th day of the experiment. Afterwards, the oxidative stress and inflammatory markers, the histopathological and immunohistochemical alterations of the parotid tissues in the studied groups were evaluated. CYP intoxication induced a significant parotid tissue injury represented by the elevation in the values of malondialdehyde (MDA), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) and decrease in the catalase activity and glutathione peroxidase (GPx). Histologically, extensive histopathological alterations e.g., widely spaced serous acini with irregular shapes and congested blood vessels as well as downregulated ki-67 and alpha-smooth muscle actin (α-SMA) immunoexpression were induced by CYP. HSP administration markedly improved the biochemical and the histopathological studies. We can conclude that HSP elicited protective effects against the CYP-induced parotid toxicity.

Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

Rationale: Trauma, surgery, and infection can cause severe inflammation. Both dysregulated inflammation intensity and duration can lead to significant tissue injuries, organ dysfunction, mortality, and morbidity. Anti-inflammatory drugs such as steroids and immunosuppressants can dampen inflammation intensity, but they derail inflammation resolution, compromise normal immunity, and have significant adverse effects. The natural inflammation regulator mesenchymal stromal cells (MSCs) have high therapeutic potential because of their unique capabilities to mitigate inflammation intensity, enhance normal immunity, and accelerate inflammation resolution and tissue healing. Furthermore, clinical studies have shown that MSCs are safe and effective. However, they are not potent enough, alone, to completely resolve severe inflammation and injuries. One approach to boost the potency of MSCs is to combine them with synergistic agents. We hypothesized that alpha-1 antitrypsin (A1AT), a plasma protein used clinically and has an excellent safety profile, was a promising candidate for synergism. Methods: This investigation examined the efficacy and synergy of MSCs and A1AT to mitigate inflammation and promote resolution, using in vitro inflammatory assay and in vivo mouse acute lung injury model. The in vitro assay measured cytokine releases, inflammatory pathways, reactive oxygen species (ROS), and neutrophil extracellular traps (NETs) production by neutrophils and phagocytosis in different immune cell lines. The in vivo model monitored inflammation resolution, tissue healing, and animal survival. Results: We found that the combination of MSCs and A1AT was much more effective than each component alone in i) modulating cytokine releases and inflammatory pathways, ii) inhibiting ROS and NETs production by neutrophils, iii) enhancing phagocytosis and, iv) promoting inflammation resolution, tissue healing, and animal survival. Conclusion: These results support the combined use of MSCs, and A1AT is a promising approach for managing severe, acute inflammation.

Pulegone ameliorates inflammation and oxidative stress in L-arginine-induced acute pancreatitis in mice by regulating the activation of p38 MAPK pathway

Purpose: To evaluate the effects of pulegone (PLG) on inflammation and oxidative stress in L-arginineinduced acute pancreatitis (AP), and determine its molecular mechanism.Methods: Hematoxylin-Eosin (H &amp; E) staining assay were performed, and histopathological score, blood glucose concentration, and serum amylase level were evaluated in order to assess the effects of PLG on pancreatic injury in L-arginine-induced AP mice. Enzyme linked immunosorbent assay (ELISA) was conducted to assess the levels of myeloperoxidase (MPO), malonaldehyde (MDA) and inflammatory factors (IL-6, TNF-α and IL-1β) in L-arginine-induced AP mice. Serum lactate dehydrogenase (LDH) and relative levels of ROS generation in L-arginine-induced AP mice were determined using an LDH kit and immunofluorescence assay, respectively. The effect of pulegone (PLG) on the activation of p38 MAPK pathway in L-arginine-induced AP mice was evaluated by Western blot.Results: Significant pancreatic tissue injury occurred in L-arginine-induced AP mice was revealed. PLG alleviated the pathological injury of pancreatitis, and decreased the blood glucose concentration and serum amylase level in L-arginine-induced AP mice. In addition, PLG inhibited oxidative stress and inflammatory responses, and was enabled to inhibit the activation of p38 MAPK pathway in L-arginineinduced AP mice. Furthermore, PLG exhibited protective effect against the development of pancreatic injury in L-arginine-induced AP mice.Conclusion: PLG ameliorates L-arginine-induced inflammation and oxidative stress in AP mice in vivo by inhibiting p38 MAPK pathway, and therefore, is a potential therapeutic agent for the management of acute pancreatitis.

Genome-Wide Cell-Free DNA Methylation Analysis Reveals Significant Recipient Tissue Injury in Allograft Rejection

<h3>Purpose</h3> Acute rejection (AR) causes significant allograft injury. However, its systemic effect on recipient tissues remains poorly defined. Donor-derived cell-free DNA (ddcfDNA) is a sensitive marker of allograft rejection, but does not capture any concurrent recipient tissue injury. Herein, we leverage cfDNA epigenetics as marker of tissue injury to profile recipient tissue injury associated with AR. <h3>Methods</h3> cfDNA was extracted in 92 plasma samples from 33 heart transplant recipients (HTR) (19 with AR and 14 stable controls) from the GRAfT study (NCT 02423070), and 19 healthy controls (HC). <b>Measurements:</b> Total cell-free nuclear (ncfDNA) and mitochondrial DNA (mtcfDNA) were assayed via digital-droplet PCR. To quantify recipient cfDNA, we first measured ddcfDNA fraction by shotgun sequencing; recipient cfDNA is the difference of total ncfDNA and ddcfDNA. Next, we performed whole-genome bisulfite sequencing on cfDNA and used tissue-specific DNA methylation signatures to deconvolute recipient tissue sources of cfDNA. Median [interquartile range (IQR)] of cfDNA percent or copies (cp)/mL are reported. <h3>Results</h3> ddcfDNA levels increased 6 fold with AR compared to stable HTR (0.54 [0.32-1.21] % vs. 0.09 [0.03 - 0.12] %, <i>p</i><0.01). ncfDNA, which is >99% recipient-derived, was 5X higher in stable HTR (7997 [3974-9740] cp/mL) than HC (1224 [921-1985] cp/mL, <i>p</i><0.01). With AR, ncfDNA doubled (14949 [8460-24018] cp/mL) compared with stable HTR (<i>p</i><0.01). mtcfDNA tripled with AR than stable HTR, and showed excellent discriminatory performance (AUC = 0.92, 95%CI [0.84-0.99]). Genome-wide methylation analysis showed significantly higher cfDNA from hematopoietic cells, vascular endothelium, hepatocyte and kidney with AR than Stable HTR, Fig.1. <h3>Conclusion</h3> Genome-wide cfDNA methylome analysis shows increased recipient tissue injury with AR. Ongoing studies intend to validate this finding and test if this approach can distinguish antibody-mediated from acute cellular rejection phenotypes.

Commentary: Cryotherapy and the sebaceous glands.

Sebaceous glands are exocrine epithelial glands secreting protective surface lipids via a holocrine mechanism in which the sebocytes undergo a programmed disintegration. Sebaceous glands are found in the eyelids as the tarsal meibomian glands and the glands of Zeis near eyelash hair follicles.[1] Pathologically, the ophthalmic sebaceous gland lesions range from benign cysts and chalazia to malignant sebaceous gland carcinoma (SGC), which happens to be the most common eyelid malignancy in the Indian population.[2] Cryotherapy has been extensively used in ophthalmology, dermatology, and surgical specialties as a primary or adjunctive treatment for lesions ranging from intraocular tumors, eyelid cysts, skin tags, warts, and acne vulgaris to cervical and hepatic cancers.[3] There are two mechanisms of cryotherapy: (a) freezing-related direct cellular injury and (b) vascular disruption of the lesion. Its role in the treatment of localized basal cell carcinomas of the eyelid has been documented in the literature with a low recurrence rate.[2] The prolonged healing time with secondary intention healing and depigmentation remains a concern with cryotherapy of the skin tumors. However, it helps in preserving the functionality of essential structures like eyelids and the surrounding lacrimal system. The in vivo biological effect of cryotherapy on normal sebaceous glands has been evaluated with the help of coherent anti-Stokes Raman scattering microscopy by Jung et al.[4] They demonstrated that the effect of lipid crystallization had a limited role in causing cellular disruption. Instead, sebocyte loss occurred at temperatures lower than those required for lipid crystallization. In addition, the cryo-treated sebaceous glands began recovering 1–2 weeks after treatment. Ray Jalian et al.[5] also investigated the role of controlled local skin cooling in causing preferential injury to sebaceous glands to understand its mechanism in treating acne vulgaris. They observed that cooling-induced damage led to a 20% reduction in sebum output for 2 weeks, with minimal collateral injury to surrounding tissues. A higher number of freeze–thaw cycles and slower thawing resulted in more significant tissue injury. Immunohistochemistry-based expression of Ki67 (a proliferative marker) and keratin 15 (a progenitor basal cell marker) was not disrupted by cooling. This led them to conclude that cooling-induced damage was temporary and occurred due to disruption of cellular architecture, enzymatic activity, and decreased lipid content. The sebum output recovered after 4 weeks. Though both these studies used a temperature higher than what is used in ophthalmic practice, they did conclusively demonstrate the limited level of cytotoxicity as well as the temporary duration of the effect of cryotherapy on sebaceous glands. The ophthalmic literature describes the use of cryotherapy as an adjunct in addressing the pagetoid spread seen in eyelid SGC.[6] Lisman (1989) first reported the use of cryotherapy to manage the map biopsy-proven sites of pagetoid spread in six cases of sebaceous gland carcinoma, thereby avoiding exenteration.[7] Shields et al.[6] advocated the application of triple freeze cryotherapy on conjunctival edges to eliminate residual tumor cells. On the contrary, Esmaeli et al.[8] reported satisfactory outcomes in 50 patients of SGC without the need for cryotherapy. A survey of practice patterns in Nordic countries revealed that only 27% of respondents offered cryotherapy as an adjunctive treatment for addressing the pagetoid spread in SGC.[9] Thus, the biological effect of cryotherapy on sebaceous glands is limited and reversible. The reassurance provided by triple freeze and slow thaw to the margins may not be reliable in clinical practice. This fact has been highlighted by Alam et al.[10] in the present issue of this journal. They performed a histopathological study on a portion of excised SGC tumor that was subjected to cryotherapy. Greater than 50% of viable tumor cells along with extensive areas of necrosis were observed on microscopy, and the authors concluded that the cell damage induced by cryotherapy is insufficient for achieving adequate tumor control. We believe that an early diagnosis and timely treatment using a wide (~3-mm margin) full-thickness excision, intraoperative frozen section control, and map biopsy for pagetoid spread remains the preferred practice pattern. The adjunctive use of cryotherapy remains doubtful in the SGC of eyelids. Close monitoring and regular follow-up to observe for recurrences are vital in the long-term management of patients.

LUNG PROTECTIVE POTENTIAL EFFECT OF ZILEUTON DURING ENDOTOXAEMIA MODEL IN MALE MICE.

The aim: This study was undertaken to investigatethe possible lung protective potential effect of zileuton during polymicrobial sepsis, through modulation of inflammatory and oxidative stress pathway. Materials and methods: 24 adult male Swiss-albino mice aged 8-12 weeks, with a weight of 25-35g, were randomized into 4 equal groups n=6, sham (laparotomy without CLP), CLP (laparotomy with CLP), vehicle (equivalent volume of DMSO 1 hour prior to CLP), and Zileuton (5 mg/kg 1 hour prior to CLP) group. After 24 hrs. of sepsis, the lung tissue harvested and used to assess IL-6, IL-1B, IL-17, LTB-4,12(S) HETE and F2-isoprostane as well as histological examination. Results: Lung tissue inflammatory mediators IL-6, IL-1B, IL-17, LTB, 12 (S) HETE) and oxidative stress were carried out via ELISA. Lung tissue levels of IL-6, IL-1B, IL-17, LTB4, 12(S) HETE and oxidative stress (F2 isoprostan)level were significantly higher in sepsis group (p<0.05) as compared with sham group, while zileuton combination showed significant (p<0.05) lower level in these inflammatory mediators and oxidative stress as comparedto sepsis group. Histologically, All mice in sepsis group showed a significant (p<0.05) lung tissue injury, while in zileuton pretreated group showed significantly (p<0.05) reduced lung tissue injury. Conclusions: The results of the present study revealed that zileuton has the ability to attenuate lung dysfunction during CLP induced polymicrobial sepsis in male mice through their modulating effects on LTB4,12(S) HETE and oxidative stress downstream signaling pathways and subsequently decreased lungtissue levelsof proinflammatory cytokines (IL-1β, and IL-6,IL-17).

Novel Pressure-Regulated Deployment Strategy for Improving the Safety and Efficacy of Balloon-Expandable Transcatheter Aortic Valves.

The authors propose a novel pressure-regulated method for balloon-expandable transcatheter heart valve (THV) deployment, aimed at optimizing prosthesis-annulus apposition while preventing significant tissue injury. The optimal method for balloon-expandable THV deployment remains debated. Current protocols are volume dependent, relying on under- and overfilling of the deployment apparatus. During deployment, the annular wall tension exerted by the expanding prosthesis is determined by maximal THV diameter and balloon pressure (Laplace's law). Three hundred thirty consecutive patients with severe native aortic stenosis who underwent TAVR with SAPIEN 3 THVs were included. One hundred and six patients were considered at high risk for annular rupture. THVs were deployed until reaching a predetermined balloon pressure. Postdilatation was performed to reduce mild or greater angiographic paravalvular regurgitation (PVR). Using a biomechanical model, annular wall stress was estimated for each case and assessed against rates of postdilatation, mild or greater PVR on transthoracic echocardiography, new permanent pacemaker placement or left bundle branch block, and annular rupture. Patients with wall stress >3 MPa had reduced postdilatation rate (P< 0.001) and reduced final PVR (P=0.014). Annular rupture occurred in 2 of 3 high-risk patients with wall stress >3.5 MPa (3.69 and 3.84 MPa); no rupture occurred in 95 high-risk patients with wall stress≤3.5 MPa. We defined a single target deployment pressure per THV size to ensure deployment within target wall stress levels of 3 to 3.5 MPa: 6.25 atm for 23-mm THVs, 5.5 atm for 26-mm THVs, and 5 atm for 29-mm THVs. Patients within this target range (n=136) had a 10.0% postdilatation rate, 12.7% mild PVR, and no moderate to severe PVR. The relationship between balloon filling volume and associated pressure and wall stress was inconsistent. Pressure-regulated THV deployment is a simple, reproducible, safe, and effective method, regardless of high-risk anatomical complexities.

Novel pressure-regulated deployment strategy for improving the safety and efficacy of balloon-expandable transcatheter aortic valves

Abstract Background The optimal method for balloon-expandable transcatheter heart valve (THV) deployment remains unknown. Current implantation protocols are volume-dependent and rely on ad-hoc filling of the deployment apparatus without accounting for annular wall tension during prosthesis expansion, predisposing patients to inconsistent clinical outcomes. During THV deployment, the annular wall tension exerted by the expanding prosthesis is determined by prosthesis diameter and balloon pressure (Laplace's Law). Objective We proposed and tested a novel method for balloon-expandable THV deployment, aimed at controlling balloon pressure and the resulting annular wall tension to allow optimal prosthesis-annulus apposition while preventing significant tissue injury. Methods 330 consecutive patients with severe native aortic stenosis who underwent balloon-expandable THV implantation between 2015–2020 were included. 106 patients were considered high-risk for annular rupture. THVs were deployed until reaching a pre-determined balloon pressure; 4–4.5atm in earlier cases to establish experience and safety, later increasing to 5–6.5atm in most cases. Post-dilatation was performed to reduce &amp;gt;mild angiographic regurgitation (PVR). Using a biomechanical model, annular wall stress (tension) was estimated for each case and assessed against recorded rates of post-dilatation, ≥mild paravalvular regurgitation (PVR) on TTE, new PPM or LBBB and annular rupture. Results Patients with wall stress &amp;gt;3MPa (n=184) had reduced post-dilatation rate (p&amp;lt;0.001) and final PVR (≥mild, p=0.014). Annular rupture occurred in 2/3 high-risk cases with wall stress &amp;gt;3.5MPa; no rupture occurred in 102 high-risk cases with wall stress ≤3.5MPa. Based on these results, we defined target deployment wall stress levels (3–3.5MPa) and associated deployment pressure per THV size. Patients within this target range (n=136) had 8.1% new PPM, 12.5% new LBBB, 12.7% mild PVR with no cases of ≥moderate PVR. Importantly, there was an inconsistent relationship between deployment balloon volume and resulting annular wall stress. Conclusion Pressure-regulated THV deployment is a simple, easily reproducible, safe and effective method, regardless of high-risk anatomical complexities. Funding Acknowledgement Type of funding sources: None. Annular wall stress and PVRModel, stress vs volume and new strategy

Choleoeimeria pogonae Alters the Bile Acid Composition of the Central Bearded Dragon (Pogona vitticeps)

The coccidian parasite Choleoeimeria pogonae infests the biliary ducts and gallbladder of the central bearded dragon (Pogona vitticeps). Endogenous C. pogonae development occurs in the epithelium of the gallbladder and bile ducts, leading to significant tissue injury. To determine whether bile composition in the gallbladder was disrupted by the parasite, bile samples were collected from one normal and one C. pogonae–infected central bearded dragon. Bile acid species were identified and quantified with liquid chromatography and mass spectroscopy. Tauroallochoic acid was the predominant bile acid (82.2%) in the normal bearded dragon, whereas the deconjugated allocholic acid was the predominant bile acid (40.1%) in the bearded dragon with C. pogonae. Taurine conjugation inhibits calcium precipitation in bile and bile acid–mediated ductal epithelial cytotoxicity. The shift in bile acid content identified in the C. pogonae infected bearded dragon may contribute to cholelithiasis and mucosal damage.

Opioid Utilization Patterns in United States Patients with Sickle Cell Disease

Opioid Utilization Patterns in United States Patients with Sickle Cell Disease

Access to Care for Medicaid and Commercially-Insured United States Patients with Sickle Cell Disease

Access to Care for Medicaid and Commercially-Insured United States Patients with Sickle Cell Disease

MiR-210-3p Enhances Cardiomyocyte Apoptosis and Mitochondrial Dysfunction by Targeting the NDUFA4 Gene in Sepsis-Induced Myocardial Dysfunction.

Sepsis-induced myocardial dysfunction (SIMD) is a common complication with high incidence rates in sepsis patients. This study aimed to investigate the roles of miR-210-3p in regulating cardiomyocyte apoptosis and mitochondrial dysfunction associated with SIMD pathogenesis.A rat sepsis model was established by cecal ligation and puncture. Serum inflammatory factors, myocardial tissue apoptosis, and expression of miR-210-3p were evaluated. In vitro, miR-210-3p expression in H9C2 cells was altered by transfection with its mimics or inhibitors. H9C2 viability was assessed via CCK-8 assay, and reactive oxygen species (ROS) production and apoptosis were detected through flow cytometry. The targeting regulatory relations between miR-210-3p and NADH dehydrogenase (ubiquinone) 1 alpha subcomplex 4 (NDUFA4) were validated by dual luciferase reporter assay.The rat sepsis model showed increased serum TNF-α and IL-6 levels, significant myocardial tissue injuries and apoptosis with decreased Bcl-2 and increased Caspase-1 protein levels. In vitro, septic rat serum suppressed viability, promoted ROS production and apoptosis, impaired COX IV activities and increased cytochrome release in H9C2 cells. The expression of miR-210-3p was greatly increased in myocardial tissues of septic rats and septic serum-treated H9C2 cells. miR-210-3p directly binds to the 3' UTR of the NDUFA4 gene. Septic rat serum suppressed NDUFA4 and Iron-Sulfur Cluster Assembly Protein U gene expressions in H9C2 cells. The above cellular and molecular alterations in H9C2 cells induced by septic serum were enhanced by miR-210-3p mimics and abrogated by miR-210-3p inhibitors.miR-210-3p promoted SIMD pathogenesis by targeting NDUFA4 to enhance cardiomyocyte apoptosis and impair mitochondrial function.