Day 1 Research Articles

Effect of day 3 embryo cell number on the pregnancy and neonatal outcomes of day 4 single embryo transfer from fresh cycles

BackgroundThe aim of this retrospective cohort study was to assess the impact of day 3(D3) embryo cell number on the clinical pregnancy and neonatal outcomes of day 4(D4) single embryo transfer in fresh cycles.MethodsThe study included 431 day 4 single embryo transfer in fresh cycles conducted between December 2018 and June 2023. These cycles were divided into three groups according to the day 3 embryo cell number: 248 cycles in the 7 ~ 9-cell group, 149 cycles in the 10 ~ 13-cell group and 34 cycles in the >13-cell group, and clinical pregnancy outcomes and neonatal outcomes were compared among the three groups.ResultsThe clinical outcomes with 10 ~ 13-cell were significantly higher than those of the 7 ~ 9-cell group, regardless of whether the female age was < 30 or ≥ 30 years. The same result could be found when the insemination pattern was IVF, and when the transferred embryos were the grade of complete fusion. There were no differences in neonatal outcomes between different groups.After adjusting for confounding factors, the 7 ~ 9-cell group was associated with lower clinical pregnancy rates(CBR) and live birth rates(LBR) compared with the 10 ~ 13-cell group (CPR: aOR 0.527, 95% CI 0.317 ~ 0.874, P = 0.013; LBR: aOR 0.499, 95% CI 0.308 ~ 0.807, P = 0.005).ConclusionThe cell number of D3 embryos can be an important reference indicator for D4 embryo selection. When performing day 4 single embryo transfer in fresh cycles, embryos with 10 ~ 13-cell on D3 can be preferentially selected for transplantation to enhance clinical outcomes, especially when the insemination pattern is IVF, and when the transferred embryos are the grade of compaction stage.

Deciphering Mechanisms of Adipocyte Differentiation in Abdominal Fat of Broilers.

The excessive deposition of abdominal fat tissue (AFT) in broilers has emerged as a major concern in the poultry industry. Despite some progress in recent years, the molecular mechanisms underlying AFT development remain ambiguous. The current study combined RNA-seq with transposase-accessible chromatin sequencing (ATAC-seq) to map the dynamic profiling of chromatin accessibility and transcriptional reprogramming in AFT adipocyte differentiation in broilers at day 3 (D3) and D14. Our results found that the levels of CDK1 and PCNA were down-regulated at D14, D28, and D42 compared to D3, while the levels of C/EBPα and FABP4 were up-regulated at D14 and D42 compared to D3. Meanwhile, PPARγ was significantly up-regulated at D28 and D42. RNA-seq of AFT identified 1705 up-regulated and 1112 down-regulated differential expression genes (DEGs) between D3 and D14. Pathways based on up-regulated DEGs mainly enriched some pathways related to adipocyte differentiation, while down-regulated DEGs pointed to DNA replication, cell cycle, and gap junction. Gene set enrichment analysis (GSEA) revealed that DNA replication and the cell cycle were down-regulated at D14, while the insulin signaling pathway was up-regulated. In the OA-induced immortalized chicken preadipocyte (ICP2) model, protein dynamic changes were consistent with AFT from D3 to D14. Same pathways were enriched in ICP2. In addition, based on overlapped DEGs from AFT and ICP2, enriched pathways related to adipocyte differentiation or proliferation mentioned above were all involved. A total of 1600 gain and 16727 loss differential peaks (DPs) were identified in ICP2 by ATAC-seq. Predicted genes from DPs at the promoter regions were enriched in glycerophospholipid metabolism, TGF-β signaling, FoxO signaling, and ubiquitin-mediated proteolysis. DNA motifs predicted 159 transcription factors (TFs) based on gain and loss peaks from the promoter regions, where 1 and 10 TFs were overlapped with up or down TFs from DEGs. Overall, this study presents a framework for the comprehension of the epigenetic regulatory mechanisms of adipocyte differentiation and identifies candidate genes and potential TFs involved in AFT adipocyte differentiation in broilers.

Safety, tolerability and blood pressure lowering of the innovative guanylyl cyclase a-receptor activator MANP in an ethnically diverse resistant hypertension population: a phase 1B clinical trial

Abstract Background/Introduction MANP is a novel atrial natriuretic peptide (ANP) analog bioengineered as a particulate guanylyl cyclase A receptor (GC-A) activator with superior receptor binding, resistance to neprilysin degradation, natriuretic, aldosterone suppressing and blood pressure (BP) lowering properties than native ANP. MANP is now under clinical development for resistant hypertension (RH). The rationale for its design was the fundamental role of the ANP/GC-A system in BP homeostasis through which GC-A activation with MANP leads to BP reduction through renal, vascular, and hormonal mechanisms mediated by the second messenger cGMP. Purpose We investigated the safety, tolerability, cGMP activation and BP reductions of MANP compared to placebo in subjects with RH. Methods We performed a double-blind, placebo controlled randomized trial in 36 RH patients comprised of approximately 50% non-African Americans (NAA) and 50% AA. MANP was administered in 6 cohorts receiving ascending subcutaneous (SQ) doses of MANP once daily from 3 ug/kg to a maximal dose of 7 ug/kg or placebo for 5 days and then followed for 21 days after dosing. Endpoints included safety, cGMP activation and changes in systolic BP (SBP), aldosterone (Aldo) levels and glomerular filtration rate (GFR). Results MANP was safe and well tolerated. One patient in the 7 ug/kg cohort was discontinued due to symptomatic hypotension. Compared to baseline, plasma cGMP increased in all MANP groups on Day 1 and Day 5 with no change in the placebo group. The maximal SBP reduction in the MANP group on Days 1 (D1) and 5 (D5) was -17.5 (D1) and -16 mmHg (D5) with 3 ug/kg (lowest dose) and -39.5 (D1) and -36.3 mmHg (D5) with 7 ug/kg (highest dose); Changes in the Placebo group were -6.3 (D1) and -16.7 mmHg (D5). Notably, the SBP reductions in AA were greater than NAA in the 4 and 7 ug/kg cohorts. After the 1-hour timepoint on both the first and fifth days, Aldo and percentage of Aldo reduction generally remained suppressed compared to baseline. Twenty-one days after MANP dosing, SBP remained lower in three groups receiving SQ MANP (4, 4.5 and 7 ug/kg) compared to placebo in association with a preservation of GFR. There were no reported drug related serious adverse events. Conclusions This clinical trial demonstrates that once a day SQ administration of MANP in RH subjects activates cGMP, without attenuation over 5 days, and reduces SBP and preserves GFR. At higher doses of MANP, SBP reductions were greater in AA compared to NAA. MANP also demonstrated Aldo inhibiting actions compared to placebo. Importantly, MANP was overall safe and well-tolerated. Our findings support the continued clinical development of MANP as a novel GC-A activating peptide for the treatment of RH.

High-Flow Nasal Aerosol Therapy; Regional Aerosol Deposition and Airway Responsiveness.

Introduction: In normal subjects, during tidal breathing, aerosols deposit by settling in small airways. With obstructive lung disease (OLD), collapse of airways during expiration causes turbulence and increased deposition in central airways. High-flow nasal cannula (HFNC) therapy, washing out dead space, may affect deposition mechanisms and drug delivery. This study compared aerosol deposition and airway responsiveness in OLD after traditional and HFNC nebulization therapy. Methods: Twelve subjects with moderate to severe OLD participated in a two-day study. Spirometry was measured pre- and post-aerosol inhalation. On Day 1 (D1) subjects tidally inhaled radiolabeled albuterol (99mTc DTPA) by mouth via AeroTech II, (Biodex. Shirley, NY). Day 2 (D2) inhalation was via HFNC using i-AIRE (InspiRx, Inc. Somerset, NJ). The HFNC system (60 L/m) was infused by syringe pump at 50 mL/h. D2 lung deposition was monitored in real time by gamma camera to match D1. Pre and post heart rate, O2 sat, and nasopharyngeal deposition (NP) were measured. Mechanistic contributions were modeled using multiple linear regression (MLR) of deposition rate (DR µg/m) as a function of breathing frequency, airway geometry (FEV1), and parenchymal integrity (DLCO). Results: Albuterol lung depositions were matched (p = 0.13) with D1 central/peripheral (sC/P) ratios 1.99 ± 0.98. Following HFNC, peripheral deposition increased (31% ± 33%, sC/P = 1.51 ± 0.43, p = 0.01). D2/D1% change FVC increased by 16.1 ± 16.7% (p = 0.003). NP deposition averaged 333% of lung. Heart rate and O2 sat were unaffected (p = 0.31, p = 0.63 respectively). DR analysis was markedly different between D1 (R2 = 0.82) and D2 (R2 = 0.12). Conclusion: In subjects with OLD, HFNC nebulization at 60 L/min was well tolerated and increased peripheral drug delivery. Spirometry significantly improved. Systemic effects were undetected indicating limited nasal absorption. MLR demonstrated that different mechanisms of deposition govern traditional vs HFNC aerosol delivery. Breath-enhanced nebulization via HFNC may provide controllable and effective aerosol therapy in OLD.

Efficacy and tolerance of intravenous methylprednisolone pulses in children with severe bronchopulmonary dysplasia requiring respiratory support.

Bronchopulmonary dysplasia (BPD) is a chronic lung disease that affects premature babies and contributes to their morbidity. French BPD guidelines suggest the use of intravenous methylprednisolone pulses (IVMP) for those who require respiratory support (RS) following 36 weeks postmenstrual age (PMA). However, there is scant data on the efficacy and tolerance for this indication. Our objectives were to assess the efficacy and tolerance of IVMP in patients with severe BPD requiring RS. This retrospective observational study included preterm infants (<32 weeks gestational age) treated by IVMP between January 2016 and March 2021, in a single tertiary care hospital. The primary endpoint was signified by changes in the Pulmonary Severity Score (PSS). Our secondary endpoints were changes in RS and the tolerance of IVMP. Data of 14 children with BPD under RS were reviewed. Their median [IQR] gestational age at birth was (26[26-29.5] weeks). After IVMP administration, the PSS did not improve significantly from baseline day 0 (D0) (0.85[0.55-1.15]) to D7 (0.74 [0.48-1.00]) or Month 1 (1.02 [0.57-1.48]). Three children (21%) experienced a decrease in RS in the following month of IVMP. There was no negative short-term impact on growth. Five infants presented infectious complications. The development quotient was measured in 5 children at 2 years of age. In our study, IVMP did not improve PSS and it was associated with a high number of infections. Other studies are needed to better define the place of IVMP in these infants.

Can salivary cortisol be used in diagnosing adrenal insufficiency during the acute and subacute phases of traumatic brain injury?

The diagnosis of adrenal insufficiency (AI) related to traumatic brain injury (TBI) remains a challenge. We investigated the basal and low-dose adrenocorticotropic hormone (ACTH)-stimulated serum cortisol and salivary cortisol (SaC) levels and the diagnostic utility of SaC levels during 28 days following TBI. Blood samples were collected for basal levels [sequentially from day 1 (D1) to D7 and on D28)] and for peak serum cortisol and SaC responses to the low-dose ACTH stimulation test (on D1, D7, and D28). After the patient enrollment period was completed, patients were retrospectively categorized as AI or AS (adrenal sufficiency) for each day separately, based on a basal serum cortisol cut-off level of 11 µg/dL, and data analysis was performed between the groups. Thirty-seven patients and 40 healthy controls were included. Median basal serum cortisol levels were higher in patients on D1 but were similar on other days. Median basal SaC levels were higher in patients on D1 and D2 but were similar on other days. Median peak serum cortisol and SaC levels were similar on D1 but were lower in patients on D7 and D28. Median basal SaC levels were higher in the AS group than in the AI group on all days. When evaluating AI during the course of TBI, the cut-off for basal SaC levels is 0.5-0.6 µg/dL throughout the first week, except for 1.38 µg/dL on D2. SaC levels may serve as a surrogate marker for accurately reflecting circulating glucocorticoid activity.

Plasma neurofilament heavy chain is a prognostic biomarker for the development of severe epilepsy after experimental traumatic brain injury.

This study was undertaken to test whether the postinjury plasma concentration of phosphorylated neurofilament heavy chain (pNF-H), a marker of axonal injury, is a prognostic biomarker for the development of posttraumatic epilepsy. Tail vein plasma was sampled 48 h after traumatic brain injury (TBI) from 143 rats (10 naïve, 21 controls, 112 with lateral fluid percussion injury) to quantify pNF-H by enzyme-linked immunosorbent assay. During the 6th postinjury month, rats underwent 30 days of continuous video-electroencephalographic monitoring to detect unprovoked seizures and evaluate epilepsy severity. Somatomotor (composite neuroscore) and spatial memory (Morris water maze) testing and quantitative T2 magnetic resonance imaging were performed to assess comorbidities and lesion severity. Of the 112 TBI rats, 25% (28/112) developed epilepsy (TBI+) and 75% (84/112) did not (TBI-). Plasma pNF-H concentrations were higher in TBI+ rats than in TBI- rats (p < .05). Receiver operating characteristic curve analysis indicated that plasma pNF-H concentration distinguished TBI+ rats from TBI- rats (area under the curve [AUC] = .647, p < .05). Differentiation was stronger when comparing TBI+ rats exhibiting severe epilepsy (≥3 seizures/month) with all other TBI rats (AUC = .732, p < .01). Plasma pNF-H concentration on day 2 (D2) distinguished TBI+ rats with seizure clusters from other TBI rats (AUC = .732, p < .05). Higher plasma pNF-H concentration on D2 after TBI correlated with lower neuroscores on D2 (p < .001), D6 (p < .001), and D14 (p < .01). Higher pNF-H concentration on D2 correlated with greater T2 signal abnormality volume on D2 (p < .001) and D7 (p < .01) and larger cortical lesion area on D182 (p < .01). Plasma pNF-H concentration on D2 did not correlate with Morris water maze performance on D37-D39. Plasma pNF-H is a promising clinically translatable prognostic biomarker for the development of posttraumatic epilepsy with frequent seizures or seizure clusters.

Developmental and Molecular Effects of C-Type Natriuretic Peptide Supplementation in In Vitro Culture of Bovine Embryos.

The use of C-type natriuretic peptide (CNP) in the interaction with the oocyte and in the temporary postponement of spontaneous meiosis resumption has already been well described. However, its action in pre-implantation developmental-stage embryos is yet to be understood. Thus, our study aimed to detect the presence of the canonical CNP receptor (natriuretic peptide receptor, NPR2) in germinal vesicle (GV)-, metaphase II (MII)-, presumptive zygote (PZ)-, morula (MO)-, and blastocyst (BL)-stage embryos and, later, to observe possible modulations on the embryos when co-cultured with CNP. In Experiment I, we detected and quantified NPR2 on the abovementioned embryo stages. Further, in Experiment II, we intended to test different concentrations (100, 200, or 400 nM of CNP) at different times of inclusion in the in vitro culture (IVC; inclusion from the beginning, i.e., day 1, or from day 5). In Experiment III, 400 nM of CNP was used on day 1 (D1) in the IVC, which was not demonstrated to be embryotoxic, and it showed potentially promising results in the blastocyst production rate when compared to the control. Thus, we analyzed the embryonic development rates of bovine embryos (D7) and hatching kinetics (D7, D8, and D9). Subsequently, morula and blastocyst were collected and evaluated for transcript abundance of their competence and quality (apoptosis, oxidative stress, proliferation, and differentiation) and lipid metabolism. Differences with probabilities less than p < 0.05, and/or fold change (FC) > 1.5, were considered significant. We demonstrate the presence of NPR2 until the blastocyst development stage, when there was a significant decrease in membrane receptors. There was no statistical difference in the production rate after co-culture with 400 nM CNP. However, when we evaluated the abundance of morula transcripts, there was an upregulated transcription in ADCY6 (p = 0.057) and downregulated transcripts in BMP15 (p = 0.013), ACAT1 (p = 0.040), and CASP3 (p = 0.082). In addition, there was a total of 12 transcriptions in morula that presented variation FC > 1.5. In blastocysts, the treatment with CNP induced upregulation in BID, CASP3, SOX2, and HSPA5 transcripts and downregulation in BDNF, NLRP5, ELOVL1, ELOVL4, IGFBP4, and FDX1 transcripts (FC > 1.5). Thus, our study identified and quantified the presence of NPR2 in bovine pre-implantation embryos. Furthermore, 400 nM of CNP in IVC, a concentration not previously described in the literature, modulated some transcripts related to embryonic metabolism, and this was not embryotoxic morphologically.

Integrative multiomics analysis identifies key genes regulating intramuscular fat deposition during development

Intramuscular fat (IMF) content is an important indicator of livestock and poultry meat quality. Enhancing IMF deposition can significantly improve meat quality. Focusing on the core process of IMF deposition, this study used the Jingxing Yellow (JXY) chickens as a model organism and employed multi-omics approaches, including RNA-sequencing (RNA-seq), Whole-genome bisulfite sequencing (WGBS), and metabolomics, to identify the key genes influencing IMF deposition in chickens during development. The results indicated that the contents of triglycerides (TG) and phospholipids (PLIP) exhibited an upward trend. The TG content did not differ significantly between day 1 (D1) and day 7 (D7), but increased significantly after 35 days (D35) of age. The WGBS results revealed that CpG methylation was the predominant methylation type in the breast muscle tissue of JXY chickens. Integrative analysis of RNA-seq and WGBS identified 50 genes, including PLA2G4F, PALMD, PLSCR5, ARHGEF26, LUM, DCN, TNRC6B, CACNA1C, ROBO1, and MBTPS2, whose methylation levels were significantly negatively correlated with their expression levels. In addition, the combined Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of differentially-expressed metabolites (DEM) and differentially-expressed genes (DEG) converged on the glycerophospholipid metabolism pathway, which was significantly enriched in DEGs such as PLA2G4F, PLA2G15, LPIN1, MBOAT2, DGKH, AGPAT2, and CHKA, as well as DEM like glycerophosphocholine and phosphocholine. Notably, PLA2G4F was identified as a DEG by DNA methylation, suggesting that PLA2G4F could be a key candidate gene influencing IMF deposition during chicken development. These findings are expected to provide a solid theoretical foundation for improving meat quality through targeted genetic and epigenetic interventions.

Toxoplasma qPCR kinetics to guide pre-emptive treatment of toxoplasmosis after allogeneic hematopoietic stem cell transplantation.

Recent ECIL-guidelines recommend a quantitative PCR (qPCR) guided pre-emptive treatment approach to toxoplasmosis in seropositive recipients of allogeneic hematopoietic cell transplantation (allo-HCT). While qPCR might serve as a sensitive tool for early Toxoplasma detection, its role in treatment follow-up remains unknown. We analyzed the qPCR kinetics of allo-HCT recipients experiencing either Toxoplasma infection (TI, n=71) or disease (TD, n=14) in relation to different parameters. We included 85 patients with available qPCR values expressed as quantitative cycle (Cq) from four large hematological centers from 2009 to 2023, and kinetic analysis was performed in a selection of 74 patients screened at least weekly with blood qPCR. Day 0 (D0) was the day of anti-Toxoplasma treatment start or (when untreated) day of diagnosis. Time to qPCR negativity was inversely proportional to the Cq value at D0 (p=0.0063). Not reaching negativity at D10 was associated with a significantly higher mortality at D30 (p=0.023). Patients with a high D0-parasitic load and patients with TD showed slower clearance (p<0.001, p=0.032). Time to negativity was not significantly different for patients started on prophylactic vs curative doses as first-line treatment regimen (p=0.16). This study underscores the predictive value of qPCR kinetics monitoring in allo-HCT patients with toxoplasmosis. With the aforementioned risk factors, clinicians can identify patients at high-risk for worse outcome. Our results support to consider a therapeutic change or reinforcement if the parasitic load does not decrease after 10 days, supplementing existing clinical guidelines.

The Adolescent Patient Perspective on Activity Limitations After Sport-Related Concussion.

Assessment of sport-related concussion (SRC) has begun to include patient-reported outcome measures. However, understanding of which health limitations are most meaningful to adolescents after SRC is limited. To explore patient-perceived activity limitations after SRC and throughout recovery to return to play and mapped according to the International Classification of Functioning, Disability, and Health (ICF) model. Longitudinal study. Secondary school athletic training facilities. Fifty patients (41 males, 5 females, 4 sex not reported, age = 14.9 ± 3.5 years, grade = 10.2 ± 0.93 level) with a medically diagnosed SRC. The Patient Specific-Functional Scale (PSFS) was used to assess changes in the patient's condition and the effect the injury posed on their ability to perform activities. The PSFS is a self-reported assessment of health used to identify activity limitations and rate the difficulty of performing those tasks. The PSFS was administered to patients on days 3 (D3) and 10 (D10) after SRC and at return to play. Activities affected by injury were coded into common categories and themes by a 3-person research team for subsequent analysis. The coded themes were also mapped to the ICF domains, chapters, and categories. The dependent variables were the PSFS themes, number of activities endorsed, PSFS scores, ICF domains, chapters, and categories. Descriptive analyses and frequencies were reported for the dependent variables. A total of 157 different activities were identified at D3 and coded into 28 categories that fit into 6 themes: activities of daily living, cognitive and school (COG), sports and physical activity (SPA), screen time, sleep, and social. On D3, all patients (50/50) identified at least 1 activity limitation. Most related to SPA (37.6%) and COG (31.2%). Sixty percent of patients endorsed activity limitations at D10, primarily in COG (38.6%) and SPA (36.6%). All (100%) response categories were mapped to the ICF, with most (75%) fitting the activities and participation domain. Our primary findings suggest that SRC influences many facets of the lives of adolescent athletes. Specifically, adolescent athletes identified activity restrictions primarily related to physical activity and sports participation.

Effect of the Application of PGF2α Associated With Ovulation Induction in a Fixed-Time Superovulation Programme for Precocious Nellore Heifers.

This study evaluated the effect of prostaglandin F2α (PGF2α) associated with gonadotropin-releasing hormone (GnRH) for ovulation induction in precocious indicus heifers submitted to a fixed-time superovulation (SOV) programme. Precocious Nellore heifers (n = 35), aged 13 months, were subjected to the SOV protocol. On day 0 (D0), all animals received intravaginal insertion of a progesterone (P4) device along with intramuscular administration of 2 mg of oestradiol benzoate, plus 200 IU of follicle-stimulating hormone in decreasing doses, with 12-h intervals between D4 and D7, in addition to 150 μg of D-cloprostenol on D6 and device removal on D7. On D8, the donors received 10.5 μg of buserelin acetate and the treatment group received 300 μg of D-cloprostenol/PGF2α. Artificial insemination was performed 12 h and 24 h after GnRH administration using frozen semen. On D15 of the protocol (i.e., D7 after insemination), the embryos were collected and evaluated. All animals passed through the control and treatment groups. Results were evaluated by analysis of variance using an adjusted mixed-effects model (p < 0.05). There was no difference in the total number of embryos between the control and treatment groups (10.40 ± 1.52 vs. 9.60 ± 1.36; p = 0.63) or viable embryos (6.30 ± 1.22 vs. 4.30 ± 0.71). For precocious indicus heifers, treatment with PGF2α in association with GnRH did not affect embryo production in the fixed-time SOV protocol.

Perkembangan Morfologi dan Tingkah Laku Larva Ikan Tuna Sirip Kuning Yellowfin Tuna (Thunnus albacares)

The yellowfin tuna (Thunnus albacares) is a pelagic fish with distinct morphological development and behavioral traits. Understanding the early stages of morphology and behavior in yellowfin tuna larvae is crucial for successful cultivation efforts, particularly during the hatchery phase. This study aimed to observe and document the developmental progression and behaviors of yellowfin tuna larvae at the Tuna Laboratory of Balai Besar Riset Budidaya Laut and Penyuluhan Perikanan Gondol, Bali. Yellowfin tuna larvae were reared in 700 L round tanks at a density of 10,000 larvae per tank. Larval rearing was monitored over a period of 13 days, specifically on days 1 (D1), 3 (D3), 5 (D5), 7 (D7), 10 (D10) and 13 (D13) post-hatching. Morphological changes in the larvae were observed over time. On day 1, the larvae were transparent and planktonic, with closed mouths and eyes and the presence of melanophore pigments in their bodies. By day 3, the larvae had opened their mouths and eyes, becoming capable of feeding. The mouth opening measured between 0.34-0.68 mm (90°) and 0.18-0.37 mm (45°) on day 5 post-hatching. As the larvae grew, improvements in swimming ability were noted, correlating with the development of the swim bladder, tail fin, and dorsal fin.

Immunogenicity and safety of SARS-CoV-2 recombinant spike protein vaccine in South African people living with and without HIV-1 infection: A phase 2 randomised trial

BackgroundResponse data for COVID-19 vaccines in immunosuppressed individuals are typically limited to standard dosing in small populations. Adjusting number or interval of doses may impact immune responses based on HIV status. MethodsThis phase 2 randomised, observer-blinded, placebo-controlled South African study (2019nCoV-505/NCT05112848) enrolled medically stable people living with HIV (PLWH) and HIV-uninfected participants aged 18–65 years. PLWH were randomised 1:1:1 to receive NVX-CoV2373 on day 0 (D0) and either D21 (2-DoseD0/D21) or D70 (2-DoseD0/D70), or on D0, D21, and D70 (3-Dose). HIV-uninfected participants were randomised 1:1 to each 2-Dose regimen. PLWH were stratified into well-controlled and less–well-controlled subgroups. The primary immunologic endpoint included serum IgG and neutralising antibody responses (per geometric mean fold rise [GMFR] in titre and seroconversion rate) to ancestral SARS-CoV-2 at D35 (2-DoseD0/D21) and D84 (2-DoseD0/D70 and 3-Dose). The primary safety endpoints were participants with an unsolicited adverse event through D84, at D120, and at D180, or reactogenicity ≤7 days post-vaccination. ResultsOf 288 PLWH, 98, 96, and 94 were randomised into the 2-DoseD0/D21, 2-DoseD0/D70, and 3-Dose groups, respectively; 96 HIV-uninfected participants were randomised to the 2-DoseD0/D21 (n=47) or 2-DoseD0/D70 (n=49) regimens. Most (>85%) of the population were SARS-CoV-2 positive at baseline. Ancestral anti-spike IgG GMFRs in PLWH and HIV-uninfected participants, respectively, were 12·4 and 12·9 (D35) and 12·2 and 13·6 (D84). Comparable outcomes occurred across dosing regimens and in well-controlled and less–well-controlled PLWH. Microneutralization GMFRs at D84 in PLWH and HIV-uninfected participants, respectively, were: 6·9 and 10·1 (2-DoseD0/D21), 11·0 and 11·3 (2-DoseD0/D70), and 17·2 (PLWH 3-Dose). Antibody responses against BA.1 trended similar to those against the ancestral virus. Safety outcomes were comparable among PLWH and HIV-uninfected participants. ConclusionThis study demonstrated that NVX-CoV2373 produced consistent immunogenicity responses to SARS-CoV-2 among PLWH and HIV-uninfected participants, with no new safety signals. FundingNovavax, Inc.

The Endocannabinoid System of the Nervous and Gastrointestinal Systems Changes after a Subnoxious Cisplatin Dose in Male Rats

Background/Objectives: Cisplatin, a common chemotherapy agent, is well known to cause severe side effects in the gastrointestinal and nervous systems due to its toxic and pro-inflammatory effects. Although pharmacological manipulation of the endocannabinoid system (ECS) can alleviate these side effects, how chemotherapy affects the ECS components in these systems remains poorly understood. Our aim was to evaluate these changes. Methods: Male Wistar rats received cisplatin (5 mg/kg, i.p.) or saline on day 0 (D0). Immediately after, serial X-rays were taken for 24 h (D0). Body weight was recorded (D0, D1, D2 and D7) and behavioural tests were performed on D4. On D7, animals were euthanized, and gastrointestinal tissue, dorsal root ganglia (DRGs) and brain areas were collected. Expression of genes related to the ECS was assessed via Rt-PCR, while LC-MS/MS was used to analyse endocannabinoid and related N-acylethanolamine levels in tissue and plasma. Results: Animals treated with cisplatin showed a reduction in body weight. Cisplatin reduced gastric emptying during D0 and decreased MAGL gene expression in the antrum at D7. Despite cisplatin not causing mechanical or heat sensitivity, we observed ECS alterations in the prefrontal cortex (PFC) and DRGs similar to those seen in other chronic pain conditions, including an increased CB1 gene expression in L4/L5 DRGs and a decreased MAGL expression in PFC. Conclusions: A single dose of cisplatin (5 mg/kg, i.p.), subnoxious, but capable of inducing acute gastrointestinal effects, caused ECS changes in both gastrointestinal and nervous systems. Modulating the ECS could alleviate or potentially prevent chemotherapy-induced toxicity.

Neutrophil phenotype, effector functions and microbicidal activity in SARS-CoV-2-associated ARDS patients.

Critically ill patients admitted to the intensive care unit (ICU) for SARS-CoV-2-induced acute respiratory distress syndrome (ARDS) are at increased risk of bacterial and fungal secondary pulmonary infections due to acquired immune dysfunction. Given that the activity of neutrophils has not been described in these patients, we aimed to investigate the function of neutrophils at ICU admission and on Day 7 (D7) post admission. Neutrophil maturation and several functional indicators were investigated. We detected a significant decrease in reactive oxygen species production at D7, but we did not observe any other significant alterations in neutrophil function. Furthermore, bronchoalveolar lavage obtained from patients displayed no inhibitory effect on the function of neutrophils from healthy donors. These findings indicate that patients admitted to the ICU for SARS-CoV-2-induced ARDS do not acquire neutrophil dysfunction within the first week of their stay, which suggests that nosocomial infections among these patients are not due to acquired neutrophil dysfunctions.

Effects of HyaRegen gel on tumour proliferation of colorectal peritoneal metastases.

Pressurized intraperitoneal aerosol chemotherapy (PIPAC) is a valuable therapeutic alternative for patients with peritoneal metastases. PIPAC uses a hyaluronic acid-based gel to reduce surgically induced adhesions. The aim of this study was to evaluate the effects of the hyaluronic acid-based gel on tumor dissemination. First, we explored whether the survival of CT26 luciferase-expressing murine colonic tumor cells was correlated with the dose of HyaRegen® Gel, and we determined the half-maximal inhibitory concentration (the IC50) of the gel. Next, we performed an in vitro study of cell survival rates after gel application on day 0 (D0) and day 1 (D1). Finally, we intraperitoneally administered the gel to mice with immunocompetent BALB/c colonic peritoneal metastases (on D0, D5, D10, D14, and D18). Tumor growth was regularly monitored using a bioluminescence assay (on D11, D17, and D21). After all mice had been sacrificed on D21, the body weights and the volumes of intraperitoneal ascites were measured; the Peritoneal Carcinosis Index (PCI) and Ki-antigen 67 scores were calculated. The IC50 value was 70 μL of gel in a total volume of 100 μL. The cell survival rates on D4 were identical in the control group and the two groups that had been treated with gel on D0 and D1. The bioluminescence levels over time were similar in the gel and control groups. The PCI scores were 35.5 ± 2.89 for the control group and 36 ± 2.45 for the gel group (p = 0.8005). The mean Ki-67 index percentages were 37.28 ±1 1.75 for the control group and 34.03 ± 8.62 for the gel group (p = 0.1971). This in vitro and in vivo study using a mouse model of immunocompetent metastatic peritoneal cancer did not reveal any pro- or anti-tumoral effect of HyaRegen® Gel. These findings indicate that the gel can be used to treat PIPACs with minimal apprehension.

Performance of applications of Ophytrium-containing mousse with or without shampoo in cats with pruritic and irritated skin: a multicentre prospective field trial.

This study aimed to evaluate the performance of a protocol in which topical products (DOUXO S3 CALM Shampoo and Mousse; Ceva Santé Animale) containing Ophytrium were applied to cats to help manage feline atopic syndrome (FAS). A total of 23 client-owned cats with a history of FAS and presenting irritated skin and pruritus were recruited for this study. The cats were either shampooed or moussed on day 0 (D0) and then moussed every 48-72 h for 3 weeks. On D0, D7 and D21, clinical signs were assessed using the validated scoring system, Scoring Feline Allergic Dermatitis (SCORFAD). Pruritus was graded by the owner using an adapted dual visual analogue scale (Pruritus Visual Analog Scale for Cats [VAScat]). Veterinarians also assessed pruritus intensity and frequency, and provided a subjective assessment of global skin condition and of improvements in each cat's condition. On D21, all questionnaires were collected from both veterinarians and owners. Among the 19 cats that completed the study, the SCORFAD and VASmax (maximum value of VAScat, either scratching or licking) scores improved by ⩾50% in 63.2% and 38.9% of animals, respectively. Mean SCORFAD values decreased significantly between D0 and D21 (from 6.2 to 2.8, P <0.05). Similarly, mean VASmax values decreased significantly between D0 and D21 (from 7.4 to 4.3, P <0.05). Overall, veterinarians assessed the improvement as satisfactory, good or excellent in 18/19 (94.7%) cases. The protocol was considered efficient and practical by 18/19 (94.7%) and 19/19 (100%) owners, respectively, and the resulting good condition of skin and coat was emphasised by 15/19 (78.9%) owners. This topical protocol with Ophytrium-containing mousse and shampoo was well tolerated. The products were effective in reducing skin irritation and discomfort quickly and significantly in cats with skin irritation and pruritus, yielding high satisfaction levels among both veterinarians and owners.

Effects of fecal microbiota transplantation on clinical outcomes and fecal microbiota of foals with diarrhea.

Diarrhea in foals can be associated with disruption of the intestinal microbiota (dysbiosis). Effective management of intestinal dysbiosis in foals has not been demonstrated. Fecal microbiota transplantation (FMT) in foals with diarrhea influences the intestinal microbiota and improves clinical and clinicopathological outcomes. Twenty-five foals <6 months of age with diarrhea and systemic inflammatory response syndrome at 3 veterinary hospitals. A prospective randomized placebo-controlled cohort study. Foals in the FMT group (n = 19) or control group (n = 9) received FMT or electrolyte solution once daily for 3 days. Fecal samples were obtained on Day 0 (D0), D1, D2, D3, and D7. Within group and between group data analyses were performed for clinical, clinicopathological, and microbiota variables. Treatment had no effect on survival (FMT 79%; control 100%, P = .3) or resolution of diarrhea (FMT 68%; control 55%, P = .4). On D3, the white blood cell count of the FMT group was lower than the control group (D3 FMT group median 6.4 g/L [5-8.3 g/L]; D3 control group median 14.3 g/L [6.7-18.9 g/L] P = .04). Heart rate reduced over time in the FMT group (D0 median 80 bpm [60-150 bpm]; D2 median 70 bpm [52-110 bpm] [P = .005]; and D3 median 64, [54-102 bpm] [P < .001]). Phylum Verrucomicrobiota, genus Akkermansia, and family Prevotellaceae were enriched in the FMT group on D1 (linear discriminate analysis > 4). In foals with diarrhea, FMT appears safe and can be associated with some clinical and microbiota changes suggestive of beneficial effect.

Impact of a package of point-of-care diagnostic tests, a clinical diagnostic algorithm and adherence training on antibiotic prescriptions for the management of non-severe acute febrile illness in primary health facilities during the COVID-19 pandemic in Burkina Faso

ObjectiveTo assess the impact of an intervention package on the prescription of antibiotic and subsequently the rate of clinical recovery for non-severe acute febrile illnesses at primary health centers.MethodsPatients over 6 months of age presenting to primary health care centres with fever or history of fever within the past 7 days were randomized to receive either the intervention package constituted of point-of-care tests including COVID-19 antigen tests, a diagnostic algorithm and training and communication packages, or the standard practice. The primary outcomes were antibiotic prescriptions at Day 0 (D0) and the clinical recovery at Day 7 (D7). Secondary outcomes were non-adherence of participants and parents/caregivers to prescriptions, health workers’ non-adherence to the algorithm, and the safety of the intervention.ResultsA total of 1098 patients were enrolled. 551 (50.2%) were randomized to receive the intervention versus 547 (49.8%) received standard care. 1054 (96.0%) completed follow-up and all of them recovered at D7 in both arms. The proportion of patients with antibiotic prescriptions at D0 were 33.2% (183/551) in the intervention arm versus 58.1% (318/547) under standard care, risk difference (RD) -24.9 (95% CI -30.6 to -19.2, p < 0.001), corresponding to one more antibiotic saved every four (95% CI: 3 to 5) consultations. This reduction was also statistically significant in children from 6 to 59 months (RD -34.5; 95% CI -41.7 to -27.3; p < 0.001), patients over 18 years (RD -35.9; 95%CI -58.5 to -13.4; p = 0.002), patients with negative malaria test (RD -46.9; 95% CI -53.9 to -39.8; p < 0.001), those with a respiratory diagnosis (RD -48.9; 95% CI -56.9 to -41.0, p < 0.001) and those not vaccinated against COVID-19 (-24.8% 95%CI -30.7 to -18.9, p-value: <0.001). A significant reduction in non-adherence to prescription by patients was reported (RD -7.1; 95% CI -10.9 to -3.3; p < 0.001).ConclusionThe intervention was associated with significant reductions of antibiotic prescriptions and non-adherence, chiefly among patients with non-malaria fever, those with respiratory symptoms and children below 5 years of age. The addition of COVID-19 testing did not have a major impact on antibiotic use at primary health centers.Trial registrationClinitrial.gov; NCT04081051 registered on 06/09/2019.