High Dose Research Articles

Peptide-Based Nanoparticles Suppress Hepatic Inflammation via Blockage of Human Antigen R.

Human antigen R (HuR), which is a mRNA-binding protein that stabilizes and regulates mRNA translation, is found to have increased expression in inflammation, cancer and other diseases, making HuR to be a promising drug target. This study reports a peptide-based nanoparticle (NP) system exhibits potent anti-inflammatory activity to ameliorate acute liver injury via the ability of peptides to inhibit the mRNA binding site of HuR and block downstream signaling. Molecular modeling provided structural evidence indicating that the peptides interact with the RNA-binding site of HuR, mainly via hydrogen-bonding and hydrophobic interactions. These peptide-based NPs can act as nanocarriers to deliver peptides into cells to compete with the mRNA binding site of HuR, evidenced by the reduction of antibody recognition to the native protein and the exhibition of anti-inflammatory activity against activated macrophage cells, with no adverse effect in vitro and in vivo. In LPS/D-GalN-induced hepatic sepsis with high dosage of LPS/GalN, administration of the NPs significantly attenuated necrosis and HuR expression, resulting in the significant improvement of animal survival rate, suggesting their therapeutic potential for hepatic inflammation and a broad range of HuR-overexpressed diseases.

Acute exposure to mercury drives changes in gene expression in Drosophila melanogaster.

We quantified the effect of acute exposure to a high dosage of inorganic mercury on gene expression in Drosophila melanogaster using RNA-sequencing of whole adult females. We found 119 genes with higher gene expression following treatment (including all 5 Drosophila metallothionine genes and a number of heat shock protein genes), and 31 with lower expression (several of which are involved in egg formation). Our results highlight biological processes and genetic pathways impacted by exposure to this toxic metal, and provide motivation for future studies to understand the genetic basis of response to mercury.

Impact of fertility levels and rice genotypes on yield, quality, and chemical properties of soil under tarai region of Uttarakhand

A field experiment was conducted during kharif season of 2019 at Crop Research Centre of G.B. Pant University of Agriculture and Technology, Pantnagar (Uttarakhand) for evaluation of six rice genotypes (IET-27263, IET-26418, IET-26420, NDR-359, PD-19 and PD-26) under varying fertility levels (Control, 100% RDF, 150% RDF). Treatments were tested in Split Plot Design with three replications keeping fertilizer levels in main plots and genotypes in sub plots. Application of fertilizer at 150% RDF resulted in significantly higher grain and straw yield over all other fertilizer levels. The rice genotype IET-27263 produced highest grain, straw and biological yield over other genotypes. Similarly, total nitrogen, phosphorus and potassium content and uptake were also found to be increased significantly up to 150% RDF. Similarly, maximum uptake of N and K in grain and straw was also recorded in genotype IET-27263. It can be concluded that application of fertilizers at higher dose up to 150% RDF is essential for obtaining higher yield of rice. The new rice genotype IET27263 was found superior to all other tested genotypes.

Food Polyphenols in Radiation-Related Diseases: The Roles and Possible Mechanisms.

As science and technology continue to evolve, the potential harm of radiation to the human body cannot be overlooked. Radiation has the capacity to inflict cellular and body-wide damage. Polyphenols are a group of naturally occurring compounds that are found in an array of plant foods. Scientific studies have demonstrated that these compounds possess noteworthy anti-radiation efficacy. Furthermore, they have been observed to be less toxic at higher doses. In the present review, we discussed the mechanisms of ionizing radiation damage and the progress in the research on the radiation resistance mechanism of polyphenol compounds, to provide guidance for the prevention and treatment of radiation related diseases. Food polyphenols can reduce the oxidative damage caused by ionizing radiation, clear free radicals, reduce DNA damage, regulate NF-KB, MAPK, JAK/STAT, Wnt and other signaling pathways, improve immune function, and have significant protective effects on radiation-induced inflammation, fibrosis, cancer and other aspects. In addition, it also has significant dual effects on radiation sensitization and radiation protection. Food polyphenols come from a wide range of sources, are abundant in daily food, and have no toxic side effects, demonstrating that food polyphenols have great advantages in preventing and treating radiation-related diseases.

Characterization of Ganciclovir Dosing for the Management of Cytomegalovirus in Solid Organ Transplant Recipients Receiving Sustained Low-Efficiency Dialysis.

The optimal dosing of intravenous ganciclovir in patients receiving sustained low-efficiency dialysis (SLED) remains unclear. The primary objective is to characterize the dosing of ganciclovir for treating and preventing cytomegalovirus (CMV) in Solid Organ Transplant Recipients receiving SLED. The secondary objective is to evaluate the safety and efficacy of the dosing practices described in this study. Retrospective review of electronic medical records from solid organ transplant recipients (SOTRs) admitted to the Medical Surgical Intensive Care Unit at the Toronto General Hospital (TGH) between November 28, 2016, and September 1, 2021, was conducted. Patients concurrently receiving ganciclovir and SLED were included. Among the 27 encounters for CMV prevention, 18 patients underwent 8-hour SLED, 6 underwent 24-hour SLED, and 3 received other SLED durations. Most patients (80%) on 8-hour SLED began ganciclovir at 2.5 mg/kg/d, whereas 80% of those on 24-hour SLED started at 5 mg/kg/d. No breakthrough viremia occurred at 5 mg/kg/d, with 1 instance at 2.5 mg/kg/d. Cytopenia rates were higher at 5 mg/kg/d (33% vs 20%). For treatment (n = 20), 16 patients underwent 8-hour SLED, 2 underwent 24-hour SLED, and 2 underwent 12-hour SLED. Most (75%) on 8-hour SLED started at 2.5 mg/kg/d, whereas all on 24-hour SLED began at 5 mg/kg/d. Viral eradication rates were 75% and 60% at 2.5 and 5 mg/kg/d, respectively, with higher cytopenia rates at 5 mg/kg/d (37.5% vs 0%). Dose adjustments were primarily in response to refractory disease or cytopenia. At our institution, ganciclovir dosing patterns suggest that for patients requiring 8-hour SLED, there is clinician comfort in using 2.5 mg/kg/d for prevention and 5 mg/kg/d for treatment. In 24-hour SLED, 5 mg/kg/d may be considered for prevention. Higher doses may be considered for CMV treatment; however, we found greater variability in the dosing practices for these patients. Further research with larger sample sizes and ganciclovir drug-level assessments is needed to optimize dosing strategies for CMV treatment.

CSF1 is expressed by the intestinal epithelial cells to regulate Mφ macrophages and maintain epithelial homeostasis and is downregulated in neonates with necrotizing enterocolitis

BackgroundColony stimulating factor 1 (CSF1) is generally expressed by immune cells in response to pro-inflammatory stimuli. The CSF1 receptor (CSFR) is activated by CSF1, and plays a key role in macrophage homeostasis. Furthermore, the CSF1R+ macrophages maintain homeostasis in the intestinal epithelium. The aim of this study was to explore the functions of CSF1-expressing and CSF1R+ macrophages in necrotizing enterocolitis (NEC), which commonly affects the ileum of neonates.MethodsIn-situ CSF1 expression in the intestines of neonates with NEC or intestinal atresia (n = 4 each) was detected by immunofluorescence staining. The CSF1 levels in the intestinal crypt-derived organoid cultures were measured by ELISA. Peripheral blood monocyte-derived Mφ macrophages were co-cultured with the organoids and stimulated with lipopolysaccharide (LPS) to mimic the inflamed state of the ileum in NEC patients.ResultsCSF1 was expressed in the intestinal epithelial cells of the fetal and neonatal samples, but suppressed in the NEC samples. Furthermore, CSF1 expression was downregulated in the intestinal crypt-derived organoids by LPS. CSF1R+ macrophages were detected near the intestinal crypts in the non-inflamed intestines but were absent in tissues obtained from pediatric NEC patients. Peripheral blood monocyte-derived macrophages promoted intestinal organoid proliferation in vitro following CSF1 stimulation. Finally, low concentrations of LPS slightly enhanced the proliferation of organoids co-cultured with the macrophages, whereas higher doses had a significant inhibitory effect.ConclusionsIntestinal epithelial cells express CSF1 to regulate the resident macrophages, maintain epithelial homeostasis, and resist infection. The abundant CSF1R+ macrophages in the fetal intestine may overexpress TNF-α upon activation of the TLR4/NF-κB pathway, resulting in epithelial damage and NEC induction.

Unraveling the Omega-3 Puzzle: Navigating Challenges and Innovations for Bone Health and Healthy Aging.

Omega-3 polyunsaturated fatty acids (ω-3 PUFAs, n-3 PUFAs), including eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and alpha-linolenic acid (ALA), are essential polyunsaturated fats primarily obtained from fatty fish and plant-based sources. Compelling evidence from preclinical and epidemiological studies consistently suggests beneficial effects of ω-3 PUFAs on bone health and healthy aging processes. However, clinical trials have yielded mixed results, with some failing to replicate these benefits seen in preclinical models. This contraindication is mainly due to challenges such as low bioavailability, potential adverse effects with higher doses, and susceptibility to oxidation of ω-3 fatty acids, hindering their clinical effectiveness. This review comprehensively discusses recent findings from a clinical perspective, along with preclinical and epidemiological studies, emphasizing the role of ω-3 PUFAs in promoting bone health and supporting healthy aging. Additionally, it explores strategies to improve ω-3 PUFA efficacy, including nanoparticle encapsulation and incorporation of specialized pro-resolving mediators (SPM) derived from DHA and EPA, to mitigate oxidation and enhance solubility, thereby improving therapeutic potential. By consolidating evidence from various studies, this review underscores current insights and future directions in leveraging ω-3 PUFAs for therapeutic applications.

High-dose colistin pharmacokinetics in critically ill patients receiving continuous renal replacement therapy

BackgroundColistin, administered as intravenous colistimethate (CMS), is still used in the critical care setting and current guidelines recommend high dosage CMS in patients undergoing continuous renal replacement therapy (CRRT). Due to the paucity of real-life data, we aimed to describe colistin pharmacokinetic/pharmacodynamic (PK/PD) profile in a cohort of critically ill patients with infections due to carbapenem-resistant (CR) bacteria undergoing CRRT.ResultsAll consecutive patients admitted to three Intensive Care Units (ICUs) of a large metropolitan University Hospital, treated with colistin for at least 48 h at the dosage of 6.75 MUI q12, after 9 MIU loading dose, and undergoing CRRT were included. After the seventh dose, patients underwent blood serial sampling during a time frame of 24 h. We included 20 patients, who had CR-Acinetobacter baumannii ventilator-associated pneumonia and were characterized by a median SAPS II and SOFA score of 41 [34.5–59.3] and 9 [6.7–11], respectively. Fifteen patients died during ICU stay and six recovered renal function. Median peak and trough colistin concentrations were 16.6 mcg/mL [14.8–20.6] and 3.9 mcg/mL [3.3–4.4], respectively. Median area under the time–concentration curve (AUC0 − 24) and average steady-state concentration (Css, avg) were 193.9 mcg h/mL [170.6–208.6] and 8.07 mcg/mL [7.1–8.7]. Probability of target attainment of colistin pharmacodynamics according to the fAUC0 − 24/MIC target ≥ 12 was 100% for MIC ≤ 2 mcg/mL and 85% for MIC = 4 mcg/ML, although exceeding the toxicity limit of Css, avg 3–4 mcg/mL.ConclusionsIn critically ill patients with CR infections undergoing CRRT, recommended CMS dosage resulted in colistin plasmatic levels above bacterial MIC90, but exceeding the safety Css, avg. limit.Trial registrationThis trial was registered in ClinicalTrials.gov on 23/07/2021 with the ID NCT04995133 (https//clinicaltrials.gov/study/NCT04995133).

The evaluation of the two-dimensional gamma passing rate efficiency for the unilateral breast cancer dosimetry using the Intensity-Modulated Radiation Therapy.

To evaluate the efficacy of the commonly used two-dimensional gamma passing rate of 3%/3mm for females with unilateral breast cancer. The cross-sectional study was conducted at Al-Andalus Private Hospital, Baghdad, Iraq, from November 2021 to April 2022 after approval from the Medical Physics and Physiology Department of Mustansiriyah University, Baghdad, and comprised female patients with unilateral breast carcinomas. All the patients were subjected to a computed tomography simulation, and data was transferred to treatment planning system Monaco 5.1 for contouring and planning. An intensity-modulated radiation therapy method was selected. The planning comprised phantom detector and linear agility accelerator for pre-treatment verification. All patients were evaluated using two alternative dosimetry methods: 3%/3mm and 3%/2mm at the threshold of 10% and 5%. The Two-dimensional gamma analysis took each transversal slice in the simulation image for comparison. Data was analysed using SPSS 24. There were 15 female patients aged 26-65 years. There was a better impact of 3%/3mm compared to 3%/2mm criterion for 5% and 10% thresholds. The proportion was 10% higher than the cutoff point of 5%. The 3%/2mm criterion should be used for plans that give a high dosage to organs at risk, while the 3 %/3 mm criterion should be used for uncomplicated situations with moderate dosages to at-risk organs.

Safety and efficacy of CRISPR-mediated genome ablation of VEGFA as a treatment for choroidal neovascularization in nonhuman primate eyes

CRISPR-based genome editing enables permanent suppression of angiogenic factors like vascular endothelial growth factor (VEGF) as potential treatment for choroidal neovascularization (CNV) – a major cause of blindness in age-related macular degeneration. We previously designed adeno-associated viral (AAV) vectors with S. pyogenes Cas 9 (SpCas9) and guide RNAs (gRNAs) to target conserved sequences in VEGFA across mouse, rhesus macaque, and human, with successful suppression of VEGF and laser-induced CNV in mice. Here, we advanced the platform to nonhuman primates and found that subretinal AAV8-SpCas9 with gRNAs targeting VEGFA may reduce VEGF and CNV severity as compared to SpCas9 without gRNAs. However, all eyes that received AAV8-SpCas9 regardless of gRNA presence developed subfoveal deposits, concentric macular rings, and outer retinal disruption that worsened at higher dose. Immunohistochemistry showed subfoveal accumulation of retinal pigment epithelial cells, collagen, and vimentin, disrupted photoreceptor structure, and retinal glial and microglial activation. Subretinal AAV8-SpCas9 triggered aqueous elevations in CCL2, but minimal systemic humoral or cellular responses against AAV8, SpCas9, or GFP reporter. Our findings suggest that CRISPR-mediated VEGFA ablation in NHP eyes may suppress VEGF and CNV, but can also lead to unexpected subretinal fibrosis, photoreceptor damage, and retinal inflammation despite minimal systemic immune responses.

The nephroprotective potential of selected synthetic compound against gentamicin induced nephrotoxicity

BackgroundNephrotoxicity, the rapid impairment of kidney function caused by harmful drugs and chemicals, affects about 20% of cases and is projected to become a leading cause of death by reactive oxygen species (ROS). Gentamicin (GM), an aminoglycoside antibiotic is one of the well know drugs/chemicals to cause nephrotoxicity both in humans and animals.MethodsA study on the effects of a synthetic phenolic compound, called 5-a, on GM-induced nephrotoxicity in male Wistar albino rats was conducted. The rats were grouped into five groups: normal control (NC), GM control (GM), positive control (GM + Dexa), treatment I (GM + 5-a 5 mg/kg) and treatment II (GM + 5-a 10 mg/kg). Throughout the experiment, the rats’ weights were monitored, and at its conclusion, their serum and kidney tissues were analyzed for renal function indicators and inflammatory markers. The study also included histopathological evaluations, molecular docking studies, blood and urine analyses for electrolyte changes, and behavioural assessments for central nervous system impact.Results2-{5-[(2-hydroxyethyl)-sulfanyl]-1,3,4-oxadiazol-2-yl} phenol (5-a) significantly protected against renal damage by reducing inflammatory markers, improving antioxidant defences, and decreasing kidney injury, particularly at higher doses. The findings suggest that compound 5-a, due to its anti-inflammatory and antioxidant properties, could be a promising therapeutic option for reducing gentamicin-induced nephrotoxicity and potentially for other kidney disorders in the future.ConclusionThese findings highlight the therapeutic effects of compound 5-a in alleviating gentamicin-induced nephrotoxicity.

Dexamethasone dose intensity does not impact outcomes in newly diagnosed multiple myeloma: a secondary SWOG analysis

AbstractDexamethasone is a key component of induction for newly diagnosed multiple myeloma (NDMM), despite common toxicities, including hyperglycemia and insomnia. In the randomized ECOG E4A03 trial, dexamethasone 40 mg once weekly was associated with lower mortality than higher doses. However, the performance of dexamethasone dose reductions below this threshold with regard to progression-free survival (PFS) and overall survival (OS) in NDMM has not been fully characterized. We conducted a secondary pooled analysis of the SWOG 0777 and SWOG 1211 studies of NDMM, which used lenalidomide and dexamethasone (Rd) alone, with or without bortezomib, and with or without elotuzumab. The planned dexamethasone intensity was 40 to 60 mg weekly in all arms. Patients were categorized into FD-DEX (full-dose dexamethasone maintained throughout induction) or LD-DEX (lowered-dose dexamethasone or discontinuation; only permitted for grade 3+ toxicities per both study protocols). Of the 541 evaluated patients, the LD-DEX group comprised 373 patients (69%). There were no differences in PFS or OS between the FD-DEX and LD-DEX groups, which were balanced in terms of age, stage, and performance status. Predictors of PFS and OS in the multivariate models were treatment arm, age ≥70 years, and thrombocytopenia. FD-DEX did not significantly improve either outcome. Our study suggests that dexamethasone dose reductions are common in multiple myeloma, even within clinical trials. Given the many toxicities and unclear benefits of dexamethasone in the era of modern treatment regimens, dexamethasone dose reduction during NDMM induction warrants further prospective studies. These trials were registered at www.clinicaltrials.gov as #NCT00644228 and NCT01668719.

Study on the combustion of toluene by acid etching assisted ultrasonic supported low content Pt catalyst

Catalytic combustion technology is an effective method for the treatment of volatile organic compounds (VOCs). However, the catalytic activity of loaded noble metal catalysts is limited by factors such as high noble metal dosage and poor stability. In this study, a method for the preparation of acid etching assisted ultrasonic loaded low-content Pt-cordierite catalysts was proposed. By comparing with the 0.5 % Pt/cordierite catalyst prepared without acid etching, it was found that the catalytic activity of toluene of the 0.5 % Pt/cordierite-HNO3 catalyst was significantly improved, and the T90 of toluene combustion was only 201 ℃, which indicated that the acid etching effectively promoted the interfacial bonding between the active component Pt and the carrier. Finally, it was confirmed by XPS that the active component Pt in the 0.5 % Pt/cordierite-HNO3 catalyst existed in multiple valence states, which effectively promoted the catalytic combustion of toluene.

Performances enhancing of supersulfated cement (SSC) using waste alkaline activators: Red mud and carbide slag

Supersulfated cement (SSC), since its invention, showed lower early-age strength gain, which has limited its practical applications. The urgent issue is how to regulate and enhance its early-age performance. In this context, solid waste materials such as bauxite residue-red mud (RM) and carbide slag residue (CS) from acetylene production by calcium carbide hydrolysis were used to improve the hydration and performance of SSC. By utilizing a range of analytical techniques such as isothermal calorimetry (IC), X-ray diffraction (XRD), backscatter scanning electron images (BSE), and thermogravimetric analysis (TGA), the hydration process, phase assemblages and microstructure of CS/RM-modified SSC were systematically investigated. It’s found that CS, as a calcareous alkaline activator, act as a regulator for adjusting the two main hydration reactions of C-A-S-H deposition and ettringite formation in SSC. The alkaline activator induces the hydration of slag and promotes the formation of hydration products. However, an excessively high dosage of carbide slag delays the primary ettringite formation reaction because it inhibits the dissolution of slag and promotes the formation of outer products. It leads to a looser and more porous paste matrix, resulting in higher expansion strain and poor strength gain at full curing ages. In contrast, red mud improves the hydration rate and enhances the hydration degree of SSC within the studied content (up to 30 wt%). With addition of red mud, SSC hydration is significantly advanced and deepened, resulting in more hydrates, particularly inner products. Red mud facilitates the depolymerization of the slag glass network and promotes the forming of nuclei and growth of hydration products. However, an excessively high red mud content would lead to a looser and more porous paste matrix due to the characteristics of red mud particle itself. An optimal red mud content has been identified as 20 wt%.

Association of Daily Doses of Buprenorphine With Urgent Health Care Utilization

Higher buprenorphine doses may benefit the increasing number of individuals using fentanyl and other synthetic opioids, but there is little empirical evidence on the efficacy of such higher doses. To examine the association between higher buprenorphine doses (above 16 mg and 24 mg) and subsequent emergency department (ED) or inpatient service use among patients diagnosed with opioid use disorder. This cross-sectional study was a retrospective analysis of health data from Optum's deidentified Clinformatics Data Mart Database from 2016 to 2021 for commercially insured individuals aged 18 years or older diagnosed with opioid use disorder (OUD). Eligible participants initiated buprenorphine after at least 90 days of enrollment and were dispensed at least a 14-day supply of buprenorphine. Data were analyzed from September 2023 through February 2024. Maximum buprenorphine dose received by a patient for 14 or more days: more than 24 mg, more than 16 mg to 24 mg, more than 8 mg to 16 mg, or 1 mg to 8 mg. Days from initiation of the maximum buprenorphine dose to an ED or inpatient visit for a behavioral health diagnosis, controlling for patient demographics, comorbid conditions, time to reaching maximum dose, buprenorphine discontinuation, and pre-buprenorphine health care utilization. A total of 35 451 individuals with an OUD diagnosis who began buprenorphine treatment were identified (mean [SD] age, 46.2 [15.1] years; 20 983 male [59.2%]; 3326 Black [9.4%], 2411 Hispanic [6.8%], 26 712 White [75.3%]). The most common dose was more than 8 mg to 16 mg daily (14 802 patients [42.9%]), with 9669 patients (27.3%) in the 1 mg to 8 mg tier, 10 329 patients (29.1%) in the 8 mg to 16 mg tier, and 651 patients (1.8%) in the tier receiving more than 24 mg. Among all patients receiving buprenorphine, 12.5% experienced an ED or inpatient visit. Survival analysis shows patients receiving doses more than 24 mg and between 16 mg to 24 mg had longer times to ED or inpatient use than patients receiving from 8 mg to 16 mg (time ratio [TR], 1.11; 95% CI, 1.02 to 1.20) and more than 24 mg (TR, 1.37; 95% CI, 1.04 to 1.81). Findings for doses above 16 mg daily were consistent for observation windows as short as 365 days (more than 24 mg: TR, 1.48; 95% CI, 1.01-2.18; more than 16 mg to 24 mg: TR, 1.19; 95% CI, 1.06-1.32). These findings contribute to the sparse empirical research regarding potential benefits of higher-dose buprenorphine treatment of individuals with OUD. Clinicians should be aware of the potential effects of higher buprenorphine doses on health care utilization while policymakers work to ensure equitable access to individuals who could potentially benefit from higher doses.

Pharmacokinetic and pharmacodynamic data from the NEOLEV1 and NEOLEV2 studies

ObjectivesTo confirm that levetiracetam (LEV) demonstrates predictable pharmacokinetics(PK) at higher doses and to study the pharmacodynamics(PD) of LEV.DesignPharmacokinetic data from the NEOLEV1 and NEOLEV2 trials were analysed using a non-linear...

Synergistic effect of samarium and PVP doped MgO nanostructures for RhB degradation and inhibition of DNA gyrase with molecular docking studies

Various concentrations (2 and 4 wt%) of Samarium (Sm3+) with a fixed amount (3 wt%) of polyvinylpyrrolidone (PVP) doped magnesium oxide (MgO) nanostructures (NSs) were synthesized via facile co-precipitation route. Trivalent dopant (Sm3+) and PVP narrowed the effective band gap of MgO and, controlled the recombination dynamics of the electron-hole pair and stabilized the nanostructures. This innovative treatment aimed to harness the potential ability of MgO to target bacterial cells and eliminate notorious dyes from wastewater. Advanced analytical (EDS and elemental mapping), crystallographic (XRD, SAED, FESEM, TEM and HR-TEM), and optical (UV–Vis and Raman) techniques were operated to probe their exclusive properties. The Sm/PVP-doped MgO displayed a pronounced catalytic activity against RhB dye in an acidic medium (96.42 %). Moreover, antibacterial activity was investigated using an agar well diffusion approach. At a high dosage, the % efficacy of pure and doped nanostructures against MDR S. aureus was 55.05, 65.64, 30.06 and 46.01, respectively. In silico docking investigation indicated that Sm/PVP-doped MgO NSs could inhibit DNA gyrase enzyme in S. aureus.

Formulation Development and Evaluation of Transdermal Patches of Apigenin.

Apigenin is a flavonoid that is abundantly present in fruits and vegetables. It has a diverse range of therapeutic properties, including cardioprotective, neuroprotective, analgesic, and cancer-preventive effects. Its safety profile, even at higher doses, makes the compound particularly valuable. Apigenin, along with excipients, was used to formulate patches using the solventcasting method on a glass mold. Seven distinct batches of patches were formulated using different combinations of polymers. The drug-polymer ratios employed in these formulations were (1:2), (1:3), (1:4), (1:4), (1:(2:8), (1:(1:9), and (1:(2:8) for both drugs. The formulated patches were evaluated and results indicate that the transdermal patches (TP5) have promising results.

Morpho-Anatomical Variations in Sisymbrium irio L. Plants Raised from Seeds Treated with γ Radiation.

This study determines the effect of γ irradiation on seed germination, growth, and morpho-anatomical traits of the Sisymbrium irio L. (London rocket) plant. Seeds irradiated with 2.5, 5, 10, 15, and 20 kGy of γ radiation showed a reduced germination percentage (13.68-56.84%) with reference to the control, showing an inverse relationship with radiation dose. Observations recorded at preflowering, flowering, and postflowering stages of plant growth showed a significant (P < 0.05%) dose-dependent decline in many growth parameters, such as root length, shoot length, shoot dry weight, number of leaves, and pods per plant, due to the radiation effect. However, root dry weight, leaf length, leaf dry weight, number of branches, and number of flowers per plant increased at the lowest dose (2.5 kGy) and then declined steadily with the increasing level of radiation. Likewise, several anatomical features (length of fibers and vessel elements, diameter of vessel elements, proportion of cortex, and vasculature in the stem) showed a consistent decrease with increasing γ irradiation in treated plants compared with the control. However, the pith area and the number of vessels per microscopic field decreased significantly with the lowest radiation dose (2.5 kGy) and then increased gradually with higher doses at each ontogenetic stage. The vulnerability factor in the control as well as treated plants increased with increasing plant age. In treated plants, vulnerability was higher under the effect of low-level radiation than in the control, but it showed an inverse relationship with the increasing level of radiation, thus being the lowest at 20 kGy radiation dose. Mesomorphy also showed an almost similar variation pattern with reference to the radiation dose.

Redundancy in Innate Immune Pathways That Promote CD8+ T-Cell Responses in AAV1 Muscle Gene Transfer.

While adeno-associated viral (AAV) vectors are successfully used in a variety of in vivo gene therapy applications, they continue to be hampered by the immune system. Here, we sought to identify innate and cytokine signaling pathways that promote CD8+ T-cell responses against the transgene product upon AAV1 vector administration to murine skeletal muscle. Eliminating just one of several pathways (including DNA sensing via TLR9, IL-1 receptor signaling, and possibly endosomal sensing of double-stranded RNA) substantially reduced the CD8+ T-cell response at lower vector doses but was surprisingly ineffective at higher doses. Using genetic, antibody-mediated, and vector engineering approaches, we show that blockade of at least two innate pathways is required to achieve an effect at higher vector doses. Concurrent blockade of IL-1R1 > MyD88 and TLR9 > MyD88 > type I IFN > IFNaR pathways was often but not always synergistic and had limited utility in preventing antibody formation against the transgene product. Further, even low-frequency CD8+ T-cell responses could eliminate transgene expression, even in MyD88- or IL-1R1-deficient animals that received a low vector dose. However, we provide evidence that CpG depletion of vector genomes and including TLR9 inhibitory sequences can synergize. When this construct was combined with the use of a muscle-specific promoter, transgene expression in muscle was sustained with minimal local or systemic CD8+ T-cell response. Thus, innate immune avoidance/blockade strategies by themselves, albeit helpful, may not be sufficient to prevent destructive cellular responses in muscle gene transfer because of the redundancy of immune-activating pathways.