Intramuscular Research Articles

Safety and immunogenicity after a 30-month boost of a subtype C ALVAC-HIV (vCP2438) vaccine prime plus bivalent subtype C gp120/MF59 vaccine boost (HVTN 100): A phase 1–2 randomized double-blind placebo-controlled trial

Induction of broad, durable immune responses is a challenge in HIV vaccine development. HVTN 100 Part A administered subtype C-containing ALVAC-HIV at months 0 and 1, and ALVAC-HIV with bivalent subtype C gp120/MF59 at months 3, 6 and 12. As IgG binding antibody and T-cell responses were similar or greater at month 12.5 vs. month 6.5, but waned by month 18, we investigated vaccine-elicited immune responses after a month 30 boost in this study, HVTN 100 Part B. From 13 September 2017 to 7 August 2018, a subgroup of vaccinees was randomized to receive intramuscular injections of ALVAC+gp120/MF59 (n = 32) or gp120/MF59 alone (n = 31) and a subgroup of placebo recipients was administered placebo (n = 7) at month 30. Primary outcomes were safety, IgG binding antibodies (bAbs) to vaccine-specific and V1V2 Env proteins and vaccine-specific CD4+ T cells at month 30.5. Secondary outcomes included neutralizing and antibody dependent cellular cytotoxicity functions and durability at months 30 and 36. Both vaccine groups had an acceptable safety profile. There were no statistically significant differences in the occurrence or level of IgG bAbs between the vaccine boost groups for any vaccine-specific or V1V2 antigens. IgG responses were higher to vaccine-matched gp120 than to V1V2. The booster vaccination restored the magnitude-breadth IgG bAb response to V1V2 antigens at month 30.5. However, it rapidly waned by month 36. CD4+ T-cell response rates to the 3 vaccine-matched Env antigens for the combined vaccine groups ranged from 37% at month 30, boosted to as high as 91% at month 30.5, and waned by month 36 to as low as 44%, with no significant differences between the vaccine boost groups. Because these responses waned after 6 months, additional strategies may be needed to maintain the durability of prime-boost vaccine regimens and to generate these or other immune responses that confer protection. Trial registration: South African National Clinical Trials Register (SANCTR number: DOH—27-0215-4796) and ClinicalTrials.gov (NCT02404311).

Impact of hepatitis B immunoglobulin mode of administration on treatment experiences of patients after liver transplantation: Results from an online survey.

Hepatitis B immunoglobulin (HBIG) in combination with a potent nucleos(t)ide analog is considered the standard of care for prophylaxis against hepatitis B virus (HBV) reinfection after liver transplantation for HBV-associated disease. To evaluate patients' satisfaction, preferences, and requirements for subcutaneous (SC), intramuscular (IM), and intravenous (IV) HBIG treatments. A self-completion, cross-sectional, online, 22-question survey was conducted to examine perceptions and satisfaction with current HBIG treatment in adults receiving HBIG treatment following liver transplantation for HBV-associated disease in France, Italy, and Turkey. Hypothetical HBIG products with different administration modes were evaluated using target product profile assessment and a conjoint (trade-off) exercise. Ninety patients were enrolled; 32%, 17%, and 51% were SC, IM, and IV HBIG users, respectively. Mean duration of treatment was 36.2 months. SC HBIG had the least negative impact on emotional well-being and social life and was perceived as the most convenient, easiest to administer, least painful, and had the highest self-rating of treatment compliance. More IM HBIG users than SC or IV HBIG users reported that administration frequency was excessive (67%, 28%, and 28%, respectively). In the target product profile assessment, 76% of patients were likely to use hypothetical SC HBIG. In the conjoint exercise, administration route, frequency, and duration were key drivers of treatment preferences. Ease, frequency, duration, and side effects of HBIG treatment administration were key drivers of treatment preferences, and SC HBIG appeared advantageous over IM and IV HBIG for administration ease, convenience, and pain. A hypothetical SC HBIG product elicited a favorable response. Patient demographics, personal preferences, and satisfaction with HBIG treatment modalities may influence long-term treatment compliance.

Safety of intramuscular tixagevimab-cilgavimab (Evusheld®) administration in patients at risk of iatrogenic haematoma due to haematological disorders.

Traditionally, intramuscular (IM) injections have been avoided in patients with haematological diseases due to the risk of iatrogenic haematoma. Tixagevimab-cilgavimab (Evusheld®) is a novel monoclonal antibody combination used as preexposure prophylaxis against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), for those at highest risk of severe infections. It is delivered as two separate IM injections (1.5 mL each). Patients with haematological disease are at higher risk for severe SARS-CoV-2 infections, which may be partially abrogated by the prophylactic inoculation of tixagevimab-cilgavimab. A combined retrospective and prospective study of patients under the haematology service at a large metropolitan hospital receiving tixagevimab-cilgavimab was conducted. Tixagevimab-cilgavimab was administered IM to all patients, with platelet and factor replacement provided according to local protocols. Patients completed a numerical pain score daily for seven days following the injection, with scores ≥4/10 prompting an ultrasound to identify iatrogenic gluteal haematomas. The study recruited 131 patients; 66 patients were thrombocytopenic, including 10 patients with a platelet count <30 × 109/L. Fourteen patients (10.7%) received a single platelet transfusion prior to tixagevimab-cilgavimab administration, while two patients received fresh frozen plasma. No gluteal haematomas were identified, and only two patients reported an initial pain score of ≥4/10. The intramuscular administration of tixagevimab-cilgavimab in patients with haematological diseases was well tolerated and was not associated with iatrogenic haematoma.

Berberine Targets PKM2 to Activate the t-PA-Induced Fibrinolytic System and Improves Thrombosis

Background: Arterial thrombosis, a condition in which thrombi form in arteries, can lead to various acute cardiovascular diseases and impact the quality of life and survival of patients. Berberine (BBR), a quaternary ammonium alkaloid, has been shown to treat these diseases. However, further exploration is needed to understand underlying mechanisms of BBR. Methods and results: Rats were administered BBR via intramuscular injection. Then, an FeCl3-coated filter paper was applied to a carotid artery to induce thrombosis. The size of the thrombus and the blood flow velocity were evaluated by carotid ultrasound. The shape of the thrombus was observed using staining and microscopy. The expression levels of mRNA and proteins were verified. Additionally, mass spectrometry and single-cell RNA sequencing analysis were conducted. The administration of BBR resulted in a significant reduction in the thrombus area and an extension of the thrombus-clogging time. Furthermore, BBR administration effectively reversed the decreasing tissue–plasminogen activator (t-PA) expression and alterations in fibrinolysis system of model group. Additionally, the expression of PKM2 was suppressed following BBR administration, and the overexpression of PKM2 inhibited t-PA expression. Conclusions: BBR ameliorates thrombosis by modulating expression of PKM2, subsequently impacting the expression of t-PA within fibrinolytic system. These preliminary findings suggest that BBR could be a potential preventive and therapeutic strategy for arterial thromboembolic diseases.

Real-world data on long-acting intramuscular maintenance therapy with cabotegravir and rilpivirine mirror Phase 3 results.

Cabotegravir, an integrase strand transfer inhibitor, and rilpivirine, an NNRTI, constitute the first long-acting (LA), injectable, two-drug ART regimen approved for the maintenance of virological suppression in persons living with HIV-1 (PLHIV). The aim of this study was to assess clinical effectiveness and tolerability of LA cabotegravir/rilpivirine in a real-world setting. We conducted a retrospective, single centre study, including all PLHIV receiving LA cabotegravir/rilpivirine as standard-of-care in our tertiary centre even if initiated in clinical trials. Between 2014 and 2022, 126 PLHIV initiated LA cabotegravir/rilpivirine. All were ART-experienced, and 98.4% had a viral load (VL) of <50 copies/mL before LA cabotegravir/rilpivirine initiation. Median BMI at cabotegravir/rilpivirine initiation was 24 IQR (23-28). During a median follow-up of 9 months IQR (7-24), 27 patients discontinued cabotegravir/rilpivirine because of virological failure, 6 for adverse events, 11 for personal reasons unrelated to treatment tolerance and 5 for other reasons. Virological failure was not associated with a higher BMI, nor with weight gain during LA intramuscular (IM) cabotegravir/rilpivirine treatment, inadequate cabotegravir and rilpivirine concentrations, VL blips or the use of oral lead-in (OLI) or not. No drug resistance-associated mutation emerged. Adverse events leading to treatment interruption were injection-site pain (n = 3) and neuropsychological side effects (n = 3). A correlation between BMI and both cabotegravir and rilpivirine concentrations at 1 month post-initiation of LA-IM cabotegravir/rilpivirine was observed, with no impact of OLI. Data from this real-world cohort of PLHIV who received cabotegravir/rilpivirine LA injections suggest that this regimen is effective and well tolerated. Virological failures were not associated with the acquisition of resistance mutations.

A Retrospective Study on Botulinum Toxin Injection in Patients With Multiple Sclerosis Related Tremor: A Treatment Option Worth Trying

Objectives Multiple sclerosis (MS)-related tremor remains a disabling problem that often responds poorly to medical treatments. This study aims to evaluate the effect of botulinum toxin type A on MS-related upper limb tremor. Methods This retrospective observational cohort study included 8 patients who received intramuscular injections of onabotulinum toxin type A for medically refractory MS-related tremor. Hospital records before and 4–6 weeks after treatment, including the Fahn-Tolosa-Marin Tremor rating scale (FTM-TRS), electrophysiological tremor analysis, neurological examination findings, and videos, were reviewed retrospectively. Results There was significant improvement after botulinum toxin injection in the FTM TRS part A score for tremor (P = 0.011), FTM TRS part B score for hand function (P = 0.011), FTM Part score for activities of daily living (P = 0.012), and FTM TRS total score for overall evaluation (P = 0.012). Electrophysiological tremor analysis revealed maximal tremor amplitudes between 700–2000 μV (before treatment) and between 300–1000 μV (after treatment). The mean tremor amplitudes before and after treatment were 1350 μV and 725 μV, respectively. No adverse reactions were observed after botulinum toxin treatment. Conclusions BoNT may improve upper limb tremor and functionality in MS-related tremor. The treatment involved administering BoNT under EMG guidance, with careful muscle selection based on clinical assessment and EMG findings. This approach aimed to optimize treatment efficacy while minimizing potential adverse effects. Further studies are warranted to confirm these findings.

Adrenal crisis precipitated by omission of regular intramuscular depo-medrone injections: the importance of counselling for adrenal insufficiency in rheumatology patients on long term exogenous steroids

Adrenal crisis precipitated by omission of regular intramuscular depo-medrone injections: the importance of counselling for adrenal insufficiency in rheumatology patients on long term exogenous steroids

Pharmacokinetics and Plasma Protein Binding of Flunixin in Rainbow Trout (Oncorhynchus mykiss).

Flunixin's pharmacokinetics, bioavailability, and plasma protein binding were examined in rainbow trout. The experiment involved 252 rainbow trout (Oncorhynchus mykiss) maintained at 12 ± 0.6°C. Flunixin was administered to rainbow trout via intravascular (IV), intramuscular (IM), and oral routes at a dosage of 2.2 mg/kg. Plasma samples were collected at times 0 (control), 0.25, 0.5, 1, 2, 4, 6, 8, 10, 12, 24, 48, 72, and 96 h. High-pressure liquid chromatography-ultraviolet was employed to quantify flunixin concentrations. The elimination half-life (t1/2ʎz) for flunixin was 8.37 h for IV, 8.68 h for IM, and 8.76 h for oral. The t1/2ʎz was similar between administration groups. The volume of distribution at a steady state and total body clearance were 55.81 mL/kg and 6.83 mL/h/kg, respectively, after IV administration. The mean peak plasma concentration was 6.24 ± 0.41 μg/mL at 4 h for oral administration and 13.98 ± 0.86 μg/mL at 2 h for IM administration. The invitro protein binding ratio of flunixin in rainbow trout plasma was 96.34 ± 2.29%. The bioavailability of flunixin after oral (25.74%) administration was lower than that after IM (66.70%) administration. Thus, developing an oral pharmaceutical formulation that can be administered with feed and has high bioavailability could enhance the therapeutic effect.

Effectiveness Of Local Cold Application, Manual Pressure, Shot Blocker And Standard Injection Procedure During Tetanus Diphtheria (Td) Vaccine Booster Immunization In Reducing Pain At The Intramuscular Injection Site Of Children Studying In Selected Schools: A Systematic Review

Effectiveness Of Local Cold Application, Manual Pressure, Shot Blocker And Standard Injection Procedure During Tetanus Diphtheria (Td) Vaccine Booster Immunization In Reducing Pain At The Intramuscular Injection Site Of Children Studying In Selected Schools: A Systematic Review

Development of a novel intramuscular liposomal injection for advanced meloxicam delivery: Preparation, characterization, in vivo pharmacokinetics, pharmacodynamics, and pain assessment in an orthopedic pain model

Pain produces several physiological, and degenerative complications. This study aimed to formulate meloxicam (MLX) in liposomes to increase solubility and deliver MLX in a controlled manner to overcome its poor aqueous solubility and relatively short t1/2 problems. Liposomes were prepared by thin film hydration followed by ultrasonication. Tests for characterizing formulations included particle size, span, entrapment efficiency, drug loading, stability, differential scanning calorimetry (DSC), Fourier transformation infrared (FT-IR) spectroscopy, morphology, in vitro release, release kinetics mathematical modeling, and an in vivo pain model in dogs undergoing orthopedic surgeries, followed by in vivo pharmacokinetics, pharmacodynamics, and pain assessment studies in comparison to the reference standard, Mobitil®. Liposomal MLX had a particle size of around 100 nm, 82 % entrapment efficiency, and 4.62 % drug loading. Stability studies, DSC, and FT-IR spectroscopy indicated that liposomes were highly stable. The formulation showed an improved in vitro controlled release pattern and an enhanced in vivo pharmacokinetic behavior as manifested by higher t1/2 and AUC0–24 and lower Cl/F in comparison to Mobitil®. The pharmacodynamics study and pain scales demonstrated liposomal MLX managed postoperative pain better than Mobitil®. In conclusion, the incorporation of MLX in liposomes increased its solubility and stability, as well as its pain management properties.

PSLBII-17 Supplementation with organic versus inorganic selenium during pregnancy to support ewe and lamb health

Abstract Selenium (Se) is essential for optimal ovine health and production, and sheep raised on Se-deficient soils can be vulnerable to Se deficiencies that can affect immune function. Organic or inorganic forms of Se supplementation can remedy these consequences. While inorganic Se is less costly, organic Se is more bioavailable and may be more efficient in providing Se to ewes and lambs. The objectives of this study were to determine how organic versus inorganic Se supplementation affects ewe Se status, Se transfer from ewes to lambs, and the antibody-mediated immune response and dermal hypersensitivity response (DHR) of ewes and their lambs. Dorset-Rideau ewes (n = 100) were enrolled in a randomized complete block design study. From gestation d (gd) 110 to postpartum d (ppd) 50, ewes were fed either a Se-deficient diet and supplemented with no Se, 0.3 mg inorganic Se, or 0.3 mg or 0.6 mg organic Se. Sera were collected on gd 110, 130, 140, and ppd 10 and 50 to determine ewe Se status. On gd 110 and 120, ewes received an intramuscular injection of 0.5 mg ovalbumin (OVA) antigen, and on gd 130 the DHR to OVA was assessed by measuring change in skinfold thickness 0, 6, 24 and 48 h post intradermal injection. To measure the transfer of Se from ewe to lamb, a subset of lambs was euthanized at ppd 10, and longissimus dorsi muscle samples were collected and analyzed for Se content. On ppd 20 and 30, lambs received an intramuscular injection of 0.5 mg hen egg white lysozyme (HEWL) antigen, and on ppd 40 the DHR to HEWL was assessed using the aforementioned protocol. Analysis of ewe sera showed a positive correlation between organic Se supplementation and greater Se concentrations (P &amp;lt; 0.01). Moreover, statistical analyses of the DHR results using a generalized least squares linear model (RStudio) indicate that Se supplementation increased the immune response compared with the non-supplemented ewes (P &amp;lt; 0.01). However, no significant differences between organic and inorganic Se supplementation were found. In lambs, greater ewe Se supplementation resulted in greater Se muscle content (P &amp;gt; 0.01). and organic Se supplementation to ewes significantly increased lamb Se muscle content compared with inorganic Se supplementation (P &amp;gt; 0.01). Finally, analyses of the lamb DHR showed that maternal organic Se supplementation improved the lamb immune response compared with the control group (P &amp;gt; 0.05). Therefore, organic Se supplementation optimally enhances ewe and lamb Se status and their adaptive immune response compared with the inorganic and control groups. This knowledge will guide optimal ewe Se-supplementation choices during pregnancy and postpartum, thus promoting lamb and ewe health.

PSV-4 Clodronate re-release in response to controlled exercise and bone stressors in juvenile horses

Abstract Concerns over the extra-label use of bisphosphonates in skeletally immature horses has extended to include its likely re-release from bone into circulation in response to stressors. Therefore, the objective of this study was to be first to quantify the re-release of clodronate in response to controlled stressors. Yearling Quarter horses (n = 32) were stratified by age, body weight (BW), sex, and initial bone optical density into four treatment groups for a 168-d trial. Treatments consisted of control (CON; n = 8), single-dose (1X; d 84; n = 8), 2-dose (2X; d 0 and 84; n = 8), and four-dose groups (4X; d 0, 42, 84, and 126; n = 8). All horses received iso-volumetric intramuscular injections of either 1.8 mg/kg BW clodronate disodium (OSPHOSÒ) or saline (placebo) on d 0, 42, 84, and 126. Horses were housed in individual stalls (3.6 m × 7.2 m). Diets were formulated to meet nutrient requirements including concentrate offered every 12-h and ad libitum coastal bermudagrass (Cynodon dactylon) hay and water. Horses were exercised 5 d/wk with a progressive workload, and maximum exercise intensity was reached on d 120, verified via heart rate and blood lactate. On d 120, blood samples were collected prior to the start of the exercise bout (pre), and 0-, 0.5-, 1-, 3-, 12-, and 24-h post exercise. Both tuber coxae of each horse were biopsied on d 168 with blood samples collected prior to (pre), and 0, 12, 24, 48, 72, 168, and 336 h post biopsy. Clodronate was quantified in plasma from exercise and biopsy samples using liquid chromatography tandem mass spectrometry. Exercise and biopsy serum were analyzed for cortisol, a marker of stress, while biopsy serum was analyzed for substance P, a pain marker. Data were analyzed using SAS PROC MIXED. Exercise did not result in clodronate re-release as there was no treatment x time interaction or time effect (P ≥ 0.44), but clodronate was dose-dependently greater in treated groups throughout the exercise collection period (P &amp;lt; 0.01). A treatment x time interaction (P ≤ 0.01) was observed for clodronate concentrations surrounding bone biopsy in which concentrations decreased at 168 h post biopsy before increasing at 336 h in 4X horses. Substance P increased over time (P &amp;lt; 0.01) following biopsy, regardless of treatment. Despite decreasing over time (P ≤ 0.01), there were similarly no treatment differences (P ≥ 0.46) nor treatment ´ time interactions (P ≥ 0.78) for cortisol following either exercise or bone insult. In conclusion, submaximal exercise did not result in re-release of clodronate; however, direct bone insult disrupted clodronate concentrations in horses receiving 4 administrations of bisphosphonate despite no treatment impact on stress or pain indicators.

Pharmacokinetics of nano- and microcrystal formulations of low solubility compounds after intramuscular injection to mice.

The aim of this study was to investigate the pharmacokinetics (PK) of poorly soluble compounds when administered intramuscularly (i.m.) as crystalline particles of different sizes. Three uncharged compounds (griseofulvin, AZ'72, and AZ'07) with varying aqueous solubility were dosed to mice at 10 and 50mg/kg as nano- and microparticle formulations. The PK of the compounds was evaluated. The smaller particles of the drugs resulted in higher maximum plasma concentration (Cmax) and area under the plasma concentration-time profile (AUC) at 50mg/kg. There was a dose-proportional increase in AUC but less than dose dose-proportional increase in Cmax. The evaluation at 10mg/kg was more complex as increased exposure for nanoparticles was only observed for griseofulvin which has the highest solubility. In addition, there was an increase in half-life with an increase in dose. This study highlights that general expectations based on in vitro dissolution (i.e. that smaller particles dissolve faster than larger particles when surrounded by liquid) do not always translate to in vivo and demonstrates the importance of understanding the physicochemical properties of the drug, the characteristics of the formulations and the microphysiology at the delivery site.

PSXII-11 Impact of pre-breeding and breeding supplementation in May-born range heifers on growth rates and reproductive performance

Abstract Our objectives were to determine the impact rumen undegradable protein (RUP) supplementation prior to and during the breeding season had on growth rates and reproductive performance in May-born yearling range heifers. This study was conducted over a 3-yr period at the University of Nebraska, Gudmundsen Sandhills Laboratory (GSL) near Whitman, NE and utilized May-born Red Angus x Simmental crossbred heifers [n = 192; weaning body weight (BW) = 200 kg ± 28 kg]. Heifers were randomly assigned by pre-breeding BW to one of two treatments: 1) received 0.908 kg of a dried distillers grain supplement (DDGS; Supp, n = 66) pre-breeding through the breeding season or 2) received no supplement (NoSupp, n = 138). Each year, treatments were initiated in July and ended in August (55 ± 3 d). Using a C-Lock Smart Feeder (Rapid City, SD) heifers had individual access to their allotted DDGS supplementation. On average, NoSupp heifers consumed 0.09 kg/d (yr 1) and 0.02 kg/d (yr 2 and 3); whereas, Supp heifers averaged 0.59, 0.74, and 0.76 kg/d in yr 1, 2, and 3, respectively. Heifer BW was collected pre-trial (May), pre-breeding (July), breeding (August), pregnancy diagnosis (October), and pre-calving (April). Body condition score (BCS) was collected at pregnancy diagnosis (October) and pre-calving (April; Yr 1 and 2). Before each breeding season, two blood samples were collected 10 d apart to determine pubertal status. Heifers were synchronized with a single 5 mL intramuscular injection of prostaglandin F2 alpha (Lutalyse, Zoetis, Parsippany, NJ) 5 d after bull turnout during a 30 d natural service breeding season. Pregnancy diagnosis was determined via rectal ultrasound in October. No differences (P &amp;gt; 0.12) in BW were noted among treatments for all measurement timepoints. Moreover, SUPP heifers had greater (P &amp;lt; 0.02) average daily gain (ADG) from pre- trial to pregnancy diagnosis compared with NoSUPP heifers. However, there was no differences (P = 0.11) in ADG from pregnancy diagnosis to pre- calving. At pregnancy diagnosis (October), a treatment x year interaction (P = 0.02) for BCS was present where Supp heifers had a greater BCS than NoSupp heifers in yr 3. Heifer BCS pre-calving was not affected (P = 0.12) by treatment. There was a greater percentage (P &amp;lt; 0.01) of SUPP heifers cycling compared with NoSUPP heifers. However, there were no differences (P &amp;gt; 0.24) in pregnancy rates, the percentage that calved in the first 21 d, calf birth BW, or subsequent calving date. Ultimately, this study implies that providing a RUP supplement prior to and during the breeding season will increase the percentage of heifers that are cycling before breeding and have a greater ADG through pregnancy diagnosis; however, supplementation will not impact other reproductive parameters or BW gains.

102 Combination and individual vaccines for bovine viral diarrhea virus and infectious bovine rhinotracheitis effects on reproductive cyclicity and immune response

Abstract Vaccines are known to impact both the immune and reproductive systems. The objective of this research was to identify immunological and cyclic differences following vaccination for Bovine Viral Diarrhea Virus (BVDV) and/or Infectious Bovine Rhinotracheitis (IBR). Brahman cows (n = 15) were administered PGF2α to regress corpus lutea on d -3. Animals (d 0) were immunized (2mL, intramuscular injection) with one of three treatments: 1) Modified Live Vaccine (MLV) for BVDV and IBR (BVD+IBR; n = 5), 2) MLV for BVDV only (BVD; n = 5), or 3) sterile saline (CON; n = 5). Blood samples (30 mL/cow) were collected (d -3, 0, 2, 4, 6, 8, 10, and 14) and peripheral blood mononuclear cells (PBMC) were isolated. PBMC were incubated with propidium iodide and antibodies for specific surface cell markers (CD4, CD8, CD25, CD14, CD86, and CD335). An Amnis FlowSight flow cytometer determined cell type percentage. BVDV and IBR antibody concentrations were analyzed in serum samples (d -3, 0, and 14). Plasma samples (d 0, 2, 4, 6, 8, 10, and 14) were evaluated for interferon (IFN)-γ, interleukin (IL)-1α, IL-1β, IL-4, IL-6, IL-8, IL-10, IL-17A, macrophage inflammatory protein (MIP)-1α, MIP-1β, IL-36RA, interferon-gamma-induced protein (IP)-10, monocyte chemoattractant protein (MCP)-1, tumor necrosis factor (TNF) α, and vascular endothelial growth factor (VEGF) A concentrations by a MagPix multiplex machine using a MILLIPLEX Bovine Cytokine/Chemokine Magnetic Bead Panel. Progesterone (d 0, 2, 4, 6, 8, 10, and 14) and estradiol (d 0) concentrations were determined by RIA. Differences in antibody titers, PBMC populations, and cytokine concentrations were analyzed as repeated measures using PROC MIXED (SASv9.4). Forty percent of BVD+IBR animals experienced abnormal estrus cycles, but no BVD or CON animals did. There was a treatment effect on BVD titer concentration with BVD animals having greater titer concentrations than CON (P = 0.02) but similar concentrations to BVD+IBR (P = 0.18). There was a treatment by time interaction (P &amp;lt; 0.0001) on IBR titers with BVD+IBR animals having greater titer concentrations on d 14 compared with BVD and CON. Treatment had no effect on leukocyte populations (P ≥ 0.1409), but all populations differed over time (P ≤ 0.0045). Antigen presenting cells (CD14+) and natural killer cells (CD335+) were affected by a treatment and time interaction (P ≤ 0.0479). BVD animals had increased CD14+ cells on d 2, 4, 8, and 14 compared with CON, but CON had a greater CD335+ cell population compared with BVD and BVD+IBR on d 10. There tended to be or was a significant treatment by day interaction for IL-1α (P = 0.08), IL-1b (P = 0.03), IL-10 (P = 0.06), IFN-γ (P = 0.05), IP-10 (P = 0.02), MIP-1α (P = 0.02), MIP-1 b (P = 0.01), TNFα (P = 0.01), and VEGFA (P = 0.03). IL-1a, IL-10, IL-1b, and MIP-1 b increased in BVD+IBR from d 0 to 4 or 6 then decreased but not in CON or BVD. Increased VEGFA was observed in CON compared with BVD and BVD+IBR animals. Overall, BVD+IBR resulted in abnormal cycles and changes in cytokine concentrations and leukocyte populations. USDA is an equal opportunity provider and employer.

Efficiency of Lidocaine Intramuscular and Intraosseous Trigger Point Injections in the Treatment of Residual Chronic Pain after Degenerative Lumbar Spinal Stenosis Decompression Surgery

Efficiency of Lidocaine Intramuscular and Intraosseous Trigger Point Injections in the Treatment of Residual Chronic Pain after Degenerative Lumbar Spinal Stenosis Decompression Surgery

348 Repeat breeding syndrome: Dexamethasone decreases the prevalence of anti-sperm antibodies in dairy cows

Abstract Cows that require greater than two inseminations to conceive are diagnosed with repeat breeding syndrome (RBS). Anti-sperm antibodies (ASA) in serum are in greater abundance among individuals exhibiting fertility challenges. In humans, treatment with glucocorticoids has been shown to decrease the prevalence of ASA and improve fertility. Therefore, the purpose of this study was to determine the impact of dexamethasone on ASA and pregnancy outcomes in RBS cattle. Holstein multiparous cows (n = 18) diagnosed as having RBS were randomly allotted to one of three treatment groups: Control (C; n = 6), low dexamethasone (LD; n = 6) and high dexamethasone (HD; n = 6). Intramuscular injections of saline (C), 0.05 mg/kg of BW of dexamethasone (LD), or 0.10 mg/kg of BW of dexamethasone (HD) were administered daily starting 4 d prior to insemination (d 0) through insemination (d 4). All treatment volumes were standardized with saline to achieve the same treatment level. Blood was collected on d 0, 4, 8 and 18, with serum extracted on d 0 through 8. Additionally, milk weights were recorded until d 34. For breeding, cows were synchronized using the Double Ovsynch protocol and only one insemination was conducted at the time of breeding. The MIXED procedure in SAS was used to determine the relationship of treatment with the repeated measures of ASA abundance and milk yield. Additionally, a non-parametric chi-square analysis was conducted in SAS using PROC GENMOD to determine the relationship of treatment with immune cell count and pregnancy outcomes. Day, Treatment and Treatment × Day were designated as fixed effects, and a protected post hoc analysis was conducted to account for multiple comparisons. Anti-sperm antibodies tended to decrease (P ≤ 0.082) as dexamethasone increased compared with C, however, there was no Treatment × Day interaction. Daily milk yields for both LD and HD decreased (P &amp;lt; 0.002) compared with C cows, but there was no interaction with day. Though there was an effect for Treatment × Day (P ≤ 0.002) for white blood cell count, percentage lymphocytes, eosinophils, and neutrophils, there was no effect of treatment (P ≥ 0.181). Therefore, interaction differences were largely driven by day (P &amp;lt; 0.001). Surprisingly, there was a trend (P = 0.069) for an increase in pregnancy rates for C cows compared with LD and HD cows. In fact, one-half of the C cows achieved pregnancy compared with just one cow for LD and HD treatments. Overall, dosage of dexamethasone did not differ for ASA or pregnancy outcomes. However, dexamethasone did decrease ASA but did not increase pregnancy rates. Based on these results, we suggest that alternative treatment measures should be explored to improve pregnancy rates in RBS cows.

Role of size, surface charge, and PEGylated lipids of lipid nanoparticles (LNPs) on intramuscular delivery of mRNA

Lipid nanoparticles (LNPs) are currently the most commonly used non-viral gene delivery system. Their physiochemical attributes, encompassing size, charge and surface modifications, significantly affect their behaviors both in vivo and in vitro. Nevertheless, the effects of these properties on the transfection and distribution of LNPs after intramuscular injection remain elusive. In this study, LNPs with varying sizes, lipid-based charges and PEGylated lipids were formulated to study their transfection and in vivo distribution. Luciferase mRNA (mLuc) was entraped in LNPs as a model nucleic acid molecule. Results indicated that smaller-sized LNPs and those with neutral potential presented superior transfection efficiency after intramuscular injection. Surprisingly, the sizes and charges did not exert a notable influence on the in vivo distribution of the LNPs. Furthermore, PEGylated lipids with shorter acyl chains contributed to enhanced transfection efficiency due to their superior cellular uptake and lysosomal escape capabilities. Notably, the mechanisms underlying cellular uptake differed among LNPs containing various types of PEGylated lipids, which was primarily attributed to the length of their acyl chain. Together, these insights underscore the pivotal role of nanoparticle characteristics and PEGylated lipids in the intramuscular route. This study not only fills crucial knowledge gaps but also provides significant directions for the effective delivery of mRNA via LNPs.Graphical

Influence of Anticholinesterase Drugs on Activity and Properties of Na+,K+-ATPase in Rat Erythrocytes under Stress Caused by Intense Physical Exercise.

We studied the effect of intramuscular injection of physostigmine and neostigmine on Na+,K+-ATPase activity in erythrocytes of rats subjected to intense physical exercise. Both anticholinesterase drugs had a significant effect on the development of the stress response, which was expressed in a decrease in the manifestation of its individual components such as the concentration of ascorbic acid in the adrenal glands, stress-related erythrocyte polycythemia, and LPO indicators. Anticholinesterase drugs reverse the stress-induced decrease in Na+,K+-ATPase activity, as well as changes in its magnesium-dependent properties. There were no changes in the activity of the studied enzyme in the erythrocyte ghosts. We associate the observed differences with the correction of the functions of the cholinergic components of the hypothalamic-pituitary-adrenal axis leading to the development of a hypoergic type stress reaction.

Safety and Pharmacokinetics of Nirsevimab in Immunocompromised Children.

Immunocompromised children may have increased risk for severe respiratory syncytial virus (RSV) lower respiratory tract infection (LRTI), potentially leading to prolonged hospitalization, intensive care, and death. The open-label phase II MUSIC trial evaluated the safety and pharmacokinetics of nirsevimab, an extended half-life monoclonal antibody against RSV, in immunocompromised children aged ≤24 months. Participants received a single intramuscular injection of nirsevimab (first RSV season: 50 mg if <5 kg/100 mg if ≥5 kg; second season: 200 mg). Safety, antidrug antibodies, and pharmacokinetics were evaluated to day 361. Participants (n = 100) had ≥1 immunocompromising conditions: primary immunodeficiency (n = 33), previous transplantation (n = 16), HIV infection (n = 8) or treatment with high-dose systemic corticosteroids (n = 29), immunosuppressive chemotherapy (n = 20), or other immunosuppressive therapies (n = 15). Six children experienced eight treatment-related adverse events (none categorized as serious). Three deaths occurred, all were unrelated to treatment. Eleven children, developed antidrug antibodies, with minimal effects on pharmacokinetics and no apparent impact on safety. Nirsevimab serum concentrations at day 151 were similar to those effective in preventing medically attended RSV LRTI in healthy infants. Fourteen children had increased nirsevimab clearance. No protocol-defined medically attended RSV LRTIs occured through day 151. Among immunocompromised children aged ≤24 months, nirsevimab was well tolerated with no safety concerns and serum concentrations were supportive of efficacy. A subset of children with increased nirsevimab clearance, had conditions potentially associated with protein loss; however, the impact on efficacy is unknown.