Punicalagin attenuates myocardial oxidative damage, inflammation, and apoptosis in isoproterenol-induced myocardial infarction in rats: Biochemical, immunohistochemical, and in silico molecular docking studies.

Myocardial infarction (MI) is a life-threatening ischemic disease and is one of the leading causes of morbidity and mortality worldwide. Punicalagin (PU), the major ellagitannin found in pomegranates, is characterized by multiple antioxidant activities. The aim of this study is to assess the protective effects of PU against isoproterenol (ISO)-induced acute myocardial damage and to investigate its underlying vascular mechanisms using rat model. METHODS: Rats were randomly divided into five groups and were treated orally (p.o.) with PU (25 and 50 mg/kg) for 14 days. ISO was administered subcutaneously (S.C.) (85 mg/kg) on the 15th and 16th days to induce Myocardial infarction. Cardiac markers, oxidative stress markers, and inflammatory cytokines levels were determined in the heart tissue. Immunohistochemistry analysis was performed to determine the protein expression pathways of inflammation, apoptosis and oxidative stress (Nuclear factor erythroid 2-related factor 2 (Nrf-2), and heme oxygenase-1 (HO-1) in all the groups. In silico study was carried out to evaluate the molecular interaction of PU with some molecular targets. RESULTS: Our results showed that ISO-induced cardiac tissue injury was evidenced by increased serum creatine kinase-MB (CK-MB), cardiac troponin I (cTnI), and lactate dehydrogenase (LDH), associated with several histopathological changes. ISO also induced an increase of MDA, PCO, NO, and 8-hydroxy-2-deoxyguanosine (8-OHdG), along with a decrease of antioxidant enzyme activities in the myocardial tissues. In addition, an increase of TNF-α, NF-κB, IL-6, IL-1ß, iNOS, Nrf2 and (HO-1) was observed. Pre-treatment with PU reduced myocardial infract area, ameliorated histopathological alterations in myocardium, and decreased activities of myocardial injury marker enzymes in ISO-induced rats. In addition, PU remarkably restored ISO-induced elevation of lipid peroxidation and decrease of antioxidants, significantly reduced myocardial pro-inflammatory cytokines concentrations in this animal model. Molecular docking analysis of PU with protein targets showed potent interactions with negative binding energies. In conclusion, PU can protect the myocardium from oxidative injury, inflammatory response, and cell death induced by ISO by upregulating Nrf2/HO-1 signaling and antioxidants.

Water-Tight Arthrotomy Joint Closure of Modified Intervastus Approach in Total Knee Arthroplasty.

BACKGROUND: The joint closure technique used for total knee arthroplasty cases can have an impact on outcomes, especially when considering accelerated rehabilitation programs that follow surgery. In this study, we describe the details of the technical steps involved in performing the water-tight arthrotomy joint closure technique that we developed and use. METHODS: A total of 536 patients (average age: 62 years, average body mass index: 34 kg/m2) with primary osteoarthritis of the knee underwent total knee arthroplasty using the modified intervastus approach between 2019 and 2021. We used the water-tight arthrotomy joint closure technique to close the knee arthrotomy incision. Any infections and complications, as well as the duration of surgery and cost related to this wound closure technique, are also reported. RESULTS: Few complications were noted with this closure technique. When we first started using it, there was one case of drainage through the proximal capsular repair which required a return to the operating room 5 days postoperatively for an irrigation and debridement. We also had two cases of superficial skin necrosis along a small part of the incision line which were observed on a weekly basis and which healed uneventfully with application of betadine once daily on the necrotic area. The average time for performing wound closure after total knee arthroplasty was 45 min. CONCLUSION: We conclude that the water-tight closure approach can achieve very durable, water-tight capsule repairs and results in a decrease in postoperative wound drainage.

Hepatoprotective effect of taxifolin on cyclophosphamide-induced oxidative stress, inflammation, and apoptosis in mice: Involvement of Nrf2/HO-1 signaling.

Taxifolin (TA) is a natural flavonoid found in many foods and medicinal plants with well-documented antioxidant and anti-inflammatory properties. Cyclophosphamide (CP) is an effective antineoplastic and immunosuppressive agent; however, it is associated with numerous adverse events, including hepatotoxicity. Herein, we aimed to investigate the potential protective effects of TA using a mouse model of CP-induced hepatotoxicity. Mice were co-treated with TA (25 and 50 mg/kg, orally) and CP (30 mg/kg, i.p.) for 10 consecutive days and sacrificed 24 hours later. CP induced increased transaminases (ALT and AST), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) paralleled with pronounced histopathological alterations in the liver. Moreover, hepatic tissues of CP-injected mice showed increased malondialdehyde (MDA), protein carbonyl, and nitric oxide (NO) levels, accompanied by decreased antioxidant defenses (glutathione [GSH], superoxide dismutase [SOD], and catalase [CAT]). Livers of CP-injected mice also showed increased inflammatory response (nuclear transcription factor kappa-B [NF-κB] p65 activation, increased levels of proinflammatory cytokines tumor necrosis factor alpha [TNF-α], interleukin 1 beta [IL-1ß], and IL-6) and apoptosis (decreased Bcl-2 and increased Bax and caspase-3 expression levels). Remarkably, TA ameliorated markers of liver injury and histological damage in CP-injected mice. TA treatment also attenuated numerous markers of oxidative stress, inflammation, and apoptosis in the liver of CP-injected mice. This was accompanied by increased nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) expression in the liver tissues of CP-injected mice. Taken together, this study indicates that TA may represent a promising new avenue to prevent/treat CP-induced hepatotoxicity and perhaps other liver diseases associated with oxidative stress and inflammation.

Consequences of supplementing duck's diet with charcoal on carcass criteria, meat quality, nutritional composition, and bacterial load.

The influence of charcoal as feed additives on carcass and meat characteristics was studied in 144 four weeks old Muller ducks. The experimental ducklings were assigned to six groups of 24 birds (Eight per replicates each). The dietary treatments contained 0, 0.5, 1.0, 1.5, 2.0, and 2.5% charcoal for G1 (C), G2 (L1), G3 (L2), G4 (L3), G5 (L4) and G6 (L5), respectively. All experimental birds were raised under similar environmental and managerial conditions. Results indicated that charcoal did not affect most carcass traits significantly except for dressing percentage was higher (P < 0.05) in 1.5 and 2 % charcoal included ducks diets compared to control ducks. Charcoal supplementation significantly affected duck meat tenderness, juiciness and water holding capacity. Moreover, charcoal altered (P < 0.05) meat components such as crude protein, calcium components, desirable fatty acids, nutritional value and some bacterial counts. Thiobarbituric acid reactive substances reduced in birds fed charcoal at 1.5, 2, and 2.5%, with significant variation among treatments. No significant differences in the number of Escherichia coli and Staphylococcus aureus were detected among the ducks fed with charcoal and the control group. It could be concluded that charcoal could be included in ducks' diets at 1.5 and 2% with beneficial effects on carcass parameters.

Chemical Characterization of Clove, Basil and Peppermint Essential Oils; Evaluating Their Toxicity on the Development Stages of Two-Spotted Spider Mites Grown on Cucumber Leaves.

The two spotted spider mite (TSSM), Tetranychus urticae Koch, is a cosmopolitan mite. It rapidly reproduces and can develop resistance to chemical pesticides. This study aims to evaluate the toxicity and acaricidal activity of three essential oils from basil, clove, and peppermint against T. urticae reproduction, which is grown on three cucumber cultivars, Chief (SC 4145), Raian (CB898), and Toshka (SC 349), under laboratory conditions at 27 + 3 °C and 70 + 5% RH. GC-MS characterized the volatile oils of basil, clove, and peppermint. Methyl cinnamate, eugenol, and menthol were the main essential oils in basil, clove, and peppermint, respectively. The results indicated significant differences in the duration of development between T. urticae feeding on the three cucumber cultivars (p ≤ 0.05), including eggs, protonymph, and deutonymph time. The Toshka (SC 349) cultivar recorded the lowest developmental time. The longevity period exhibited the same trend with non-significant differences between Raian (CB898) and Toshka (SC 349). Moreover, the lethal concentration (LC50) and LC90 values in tested essential oils (EOs) showed that clove EOs were the most toxic. In contrast, basil and peppermint EOs were the least effective, and immature stages were more sensitive to EOs than adult stages. The infected Toshka (SC 349) discs treated with essential oils and abamectin under in vitro conditions indicated that clove oil is comparable to abamectin regarding its effect on the egg numbers (18.7 and 17.6 egg), immature development time, longevity, life span, and life cycle (20.6 and 20.8 days) of T. urticae. We conclude that the resistant cultivation of cucumber plants can be recommended in integrated pest management programs. The most effective of the tested oils, clove EOs, should be used as alternatives to pesticides to control T. urticae in the protected cultivation of cucumbers.

Punicalagin Protects against the Development of Methotrexate-Induced Hepatotoxicity in Mice via Activating Nrf2 Signaling and Decreasing Oxidative Stress, Inflammation, and Cell Death.

Despite its effectiveness in treating inflammatory diseases and various malignancies, methotrexate (MTX) is well known to cause hepatotoxicity, which involves increased oxidative stress and inflammation, limiting its clinical use. Herein, we looked into the effect of punicalagin (PU), a polyphenolic molecule having a variety of health-promoting attributes, on MTX-induced hepatotoxicity in mice. PU (25 and 50 mg/kg/day) was given orally to the mice for 10 days, while a single dose of MTX (20 mg/kg) was injected intraperitoneally (i.p.) at day 7. The MTX-induced liver damage was demonstrated by remarkably higher transaminases (ALT and AST), ALP, and LDH, as well as significant histological alterations in hepatic tissues. MTX-injected mice also demonstrated increases in hepatic oxidative stress markers, including malondialdehyde (MDA) and nitric oxide (NO), with a concordant drop in glutathione (GSH) content and superoxide dismutase (SOD) and catalase (CAT) activities. PU significantly attenuated the MTX-induced serum transaminases, ALP and LDH elevations, and hepatic oxidative stress measures and boosted antioxidant defenses in the liver. Moreover, the liver of MTX-treated mice showed increases in NF-κB p65 expression, pro-inflammatory cytokine (IL-6 and TNF-α) levels, and pro-apoptotic protein (caspase-3 and Bax) expression, whereas Bcl-2 and Nrf2 expressions were reduced, which were all attenuated by PU treatment. Collectively, PU inhibits oxidative damage, inflammation, and apoptosis and upregulates Nrf2 in the liver of MTX-induced mice. Thus, these findings suggest that PU may have great therapeutic potential for the prevention of MTX-induced hepatotoxicity, pending further exploration in upcoming studies.

The Kuwait Stitch: A novel surgical technique for surgical wound closures.

There are several stitch techniques used for surgical wound closing. Each stitch has its own benefits and drawbacks that a surgeon must balance before use. In this paper, we highlight some of the more common techniques utilized in operative wound closure and briefly discuss benefits and caveats one must be aware of before using. The rest of our paper will focus on a new technique, the Kuwait Stitch, with instructions on how to perform the technique followed by a discussion on the benefits and indications for use of this stitch.

Medial Tibial Plateau Stress Fracture Following Navigated Total Knee Arthroplasty: Two Case Reports.

BACKGROUND Computer navigation in total knee arthroplasty has been gaining worldwide interest among orthopedic surgeons. While there is controversial data regarding its potential better clinical outcomes compared to conventional total knee arthroplasty, it has been shown to improve component and limb alignment reliability at a potential cost of increased complications. We present 2 case reports of medial tibial stress fracture through navigated tibial cutting block pinhole sites. CASE REPORT Both cases involved morbidly obese patients who underwent a navigated total knee arthroplasty. During surgery, there were no intraoperative concerns. Both knees were well aligned postoperatively and no unusual pain was reported. At 6 months after total knee arthroplasty, a periprosthetic fracture was initiated at a cutting block pinhole site with varus collapse of the tibial component. In both cases, the pinhole site was close to the medial tibial cortex and the primary tibial component collapsed into the varus, requiring revision to a stemmed component with allograft bone. For both patients, the revision arthroplasty continues to perform well. We believe cutting block design in combination with small tibias and elevated body mass index contributed to this complication. CONCLUSIONS Robotic-assisted total knee replacement has been shown to improve precision in component alignment. We caution against placing cutting block pinholes close to the medial tibial cortex, especially in morbidly obese patients with small tibias.

Readability of Patient Educational Materials in Pediatric Orthopaedics.

BACKGROUND: Parents are increasingly turning to the internet to seek pediatric health information. Numerous organizations advise that patient educational materials (PEMs) should not surpass the sixth-grade reading level. We aimed to assess the readability of online pediatric orthopaedic PEMs. METHODS: The readability of 176 articles pertaining to pediatric orthopaedics from the American Academy of Orthopaedic Surgeons (AAOS), Pediatric Orthopaedic Society of North America (POSNA), and American Academy of Pediatrics (AAP) websites was assessed with the use of 8 readability formulae: the Flesch-Kincaid Reading Grade Level, the Flesch Reading Ease Score, the Raygor Estimate, the SMOG, the Coleman-Liau, the Fry, the FORCAST, and the Gunning Fog. The mean reading grade level (RGL) of each article was compared with the sixth and eighth-grade reading levels. The mean RGL of each website's articles also was compared. RESULTS: The cumulative mean RGL was 10.2 (range, 6.6 to 16.0). No articles (0%) were written at the sixth-grade reading level, and only 7 articles (4.0%) were written at or below the eighth-grade reading level. The mean RGL was significantly higher than the sixth-grade (95% confidence interval [CI] for the difference, 4.0 to 4.4; p < 0.001) and eighth-grade (95% CI, 2.0 to 2.4; p < 0.001) reading levels. The mean RGL of articles on the POSNA website was significantly lower than the mean RGL of the articles on the AAOS (95% CI, -1.8 to -1.0; p < 0.001) and AAP (95% CI, -2.9 to -1.1; p < 0.001) websites. CONCLUSIONS: Pediatric orthopaedic PEMs that are produced by the AAOS, the POSNA, and the AAP have readability scores that exceed recommendations. Given the increasing preference of parents and adolescents for online health information, the growing body of online PEMs, and the critical role that health literacy plays in patient outcomes, substantial work is required to address the readability of these materials.

First Reported Series of Outpatient Total Knee Arthroplasty in the Middle East.

BACKGROUND: Outpatient total knee arthroplasty (TKA) is becoming more commonplace in the United States. Alternatively, the current practice in the Middle East involves an inpatient stay of 7-10 days in the hospital after TKA. This study reports the early results of the first reported series of outpatient TKA performed on patients in the Middle East and compares the clinical and functional outcomes with those of patients who underwent inpatient TKA. METHODS: Eighty-eight patients underwent TKA (inpatient: 44 and outpatient: 44) using the modified intervastus approach in 2 hospitals in the Middle East from 2017 to 2019. Clinical and functional outcomes were assessed by recording the Knee Injury and Osteoarthritis Outcome Score, visual analog scale (VAS) for pain, and knee range of motion (ROM) preoperatively, on the day of surgery, and postoperatively at 2 weeks, 6 weeks, 3 months, and 6 months. RESULTS: Two patients undergoing outpatient TKA had complications: one patient suffered a periprosthetic fracture on postoperative day 10 after a fall, and the other patient had drainage on postoperative day 5. No complications occurred in the inpatient TKA cohort. There were no significant differences observed in the VAS scores or knee ROM numbers recorded for inpatient and outpatient TKA groups at any of the follow-up periods. Overall, the Knee Injury and Osteoarthritis Outcome Score, VAS, and ROM significantly improved 6 months after surgery compared with preoperative values for both inpatient and outpatient TKA groups. CONCLUSIONS: Outpatient TKA was safely implemented when compared with inpatient TKA, with satisfactory results. A total of 2 complications were seen in this study, which we believe are unrelated to the patient's discharge status. The concept of outpatient TKA using the modified intervastus approach was very well accepted by the patients in this study and can potentially be applied safely elsewhere in the region.

Readability of Patient Educational Materials in Hip and Knee Arthroplasty: Has a Decade Made a Difference?

BACKGROUND: The Internet has become an increasingly popular resource among orthopedic patients for health education. Numerous organisations recommend that patient educational materials (PEMs) should not exceed the 6th grade reading level. Despite this, studies have repeatedly shown the reading grade level (RGL) of PEMs to be too advanced across a range of surgical specialties. We aimed to determine the readability of online hip and knee arthroplasty PEMs. METHODS: The readability of 134 articles pertaining to hip and knee arthroplasty from 5leading worldwide healthcare websites were assessed, using 8 readability formulae; the Flesch-Kincaid Reading Grade Level, the Flesch Reading Ease Score, Raygor Estimate, SMOG, Coleman-Liau, Fry, FORCAST and Gunning Fog. The mean RGL was compared to the 6th and 8thgrade reading level. The mean RGL of each website was also compared. RESULTS: The mean cumulative RGL was 12 (range = 7-16.1). No articles (0%) were written at a 6th grade reading level and only 4 articles (3%) were written at or below the 8th grade reading level. The mean RGL was significantly higher than the 6th (95% CI, 5.62-6.30; P < .0001) and 8th grade reading level (95% CI, 3.63-4.30; P < .0001). There was a significant difference between the RGLs of the 5websites (P = .001). CONCLUSION: Arthroplasty PEMs produced by leading worldwide healthcare organisations have readability scores that are above the recommended levels. Given the imperative role of health literacy in patient outcomes and satisfaction, and the increasing prevalence of Internet use among orthopedic patients, a substantial amount of work needs to be done to improve the readability of these materials.

Thiol modified mycolic acids.

Patient serum antibodies to mycolic acids have the potential to be surrogate markers of active tuberculosis (TB) when they can be distinguished from the ubiquitously present cross-reactive antibodies to cholesterol. Mycolic acids are known to interact more strongly with antibodies present in the serum of patients with active TB than in patients with latent TB or no TB. Examples of single stereoisomers of mycolic acids with chain lengths corresponding to major homologues of those present in Mycobacterium tuberculosis have now been synthesised with a sulfur substituent on the terminal position of the α-chain; initial studies have established that one of these binds to a gold electrode surface, offering the potential to develop second generation sensors for diagnostic patient antibody detection.

Rapid magnetic resonance imaging-guided reduction of craniovertebral junction deformities.

The authors demonstrate the utility of an MR imaging-compatible traction board for the rapid reduction of craniovertebral junction (CVJ) deformities. To choose the appropriate surgical management, patients with compressive CVJ deformities often undergo a trial of traction. Conventional traction trials require the treating surgeon to infer from plain radiographs the manner in which traction forces affect neural and ligamentous structures at the CVJ. To avoid overdistraction injury, low increments of weight are added in a gradual fashion, a process that typically requires 48-72 hours. The authors outline the use of an MR imaging-compatible traction board to determine reducibility safely and rapidly in 4 patients with compressive CVJ deformities. Four patients with advanced CVJ deformities underwent a trial of MR imaging-guided traction performed using an MR imaging-compatible spine board. Serial sagittal images were acquired at baseline and following each sequential addition of force. All patients tolerated traction without neurological worsening. The neural elements were seen to be adequately decompressed in all cases during a single MR imaging session. Patients subsequently underwent craniocervical stabilization and fusion. Postoperative imaging showed maintenance of the reduction without neural compression. An MR imaging-guided trial of traction can facilitate the rapid and safe determination of the reducibility of compressive lesions in patients with advanced CVJ deformities. Rapidly acquired sagittal MR images permit the surgeon to evaluate the effects of traction on the soft tissues at the CVJ, thereby expediting the traction trial and avoiding the risks of immobility in this often-fragile patient population.

A novel magnetic approach to enhance the efficacy of cell-based gene therapies.

Attempts have been made to use various forms of cellular vectors to deliver therapeutic genes to diseased tissues like malignant tumours. However, this approach has proved problematic due to the poor uptake of these vectors by the target tissue. We have devised a novel way of using magnetic nanoparticles (MNPs) to enhance the uptake of such 'therapeutically armed' cells by tumours. Monocytes naturally migrate from the bloodstream into tumours, so attempts have been made to use them to deliver therapeutic genes to these sites. However, transfected monocytes injected systemically fail to infiltrate tumours in large numbers. Using a new in vitro assay for assessing monocyte extravasation, we show that the ability of transfected human monocytes to migrate across a human endothelial cell layer into a 3D tumour spheroid is markedly increased when cells are pre-loaded with MNPs and a magnetic force is applied close to the spheroid. Furthermore, systemic administration of such 'magnetic' monocytes to mice bearing solid tumours led to a marked increase in their extravasation into the tumour in the presence of an external magnet. This new magnetic targeting approach could be used to increase the targeting, and thus the efficacy, of many cell-based gene therapies in vivo.

Analysis of purified gp96 preparations from rat and mouse livers using 2-D gel electrophoresis and tandem mass spectrometry.

The stress protein gp96 exhibits a number of immunological activities, the majority of studies into which have used gp96 purified from a variety of tissues. On the basis of 1-D gel electrophoresis, the purity of these preparations has been reported to range between 70% and 99%. This study analyzed gp96 preparations from rat and mouse livers using 2-D gel electrophoresis and liquid chromatography electrospray ionization tandem mass spectrometry (MS-MS). The procedure for purifying gp96 was reproducible, as similar protein profiles were observed in replicate gels of gp96 preparations. The purity of the preparations was typically around 70%, with minor co-purified proteins of varying molecular weights and mobilities being present. Dominant bands at 95-100 kDa in preparations from Wistar rats and C57BL/6 mice were identified as gp96 by ECL Western blotting. Multiple bands having similar, yet distinct molecular weights and differing pI mobility on ECL Western blots were confirmed as being gp96 in preparations from Wistar rats using MS-MS. The most striking feature of the 2-D gel analysis was the presence of additional dominant bands at 55 kDa in preparations from Wistar rats, and at 75-90 kDa in preparations from C57BL/6 mice. These were identified as gp96 by ECL Western blotting and, in the case of preparations from Wistar rats, by MS-MS. Although the lower molecular weight, gp96-related molecules might be partially degraded gp96, their reproducible presence, definition and characteristics suggest that they are alternative, species-specific isoforms of the molecule. A 55 kDa protein which exhibited a lower pI value than gp96 was present in all preparations and this was identified as calreticulin, another putative immunoregulatory molecule. This study confirms the reproducibility of the gp96 purification protocol and reveals the presence of multiple gp96 isoforms, some of which likely result from post-translational modifications such as differential glycosylation and phosphorylation.

Dead or alive: immunogenicity of human melanoma cells when presented by dendritic cells.

Uveal melanoma is an aggressive malignancy with a poor prognosis despite current therapeutic intervention. These tumors have been shown to be antigenic because they express a number of melanoma-associated antigens and are therefore attractive targets for immunotherapy. Here, we investigated the immunogenicity of uveal melanoma cells that have undergone apoptosis and compared this with their necrotic or live counter-parts. The fate of the tumor antigens in these cells largely depends on their ability to be processed and phagocytosed by dendritic cells (DCs). Flow cytometric analysis shows that human DCs form conjugates more efficiently with dead uveal melanoma cells, and consequently these are effective stimuli of lymphocyte proliferation. However, only DCs pulsed with apoptotic cells were able to induce proliferation of CD8+ cytotoxic T cells and stimulate antigen-specific T cells. This study demonstrates for the first time that DCs derived from melanoma patients process and present antigens derived from both HLA-matched or HLA-mismatched human apoptotic tumor cells stimulating both CD4+ and CD8+ T cells. This approach may be important to the development of DC-based immunotherapies for melanoma.