Ml Of Phosphate-buffered Saline Research Articles

Regulatory T cells protect against diabetic cardiomyopathy in db/db mice.

Regulatory T cells (Tregs) have protected against many cardiovascular diseases. This study was intended to explore the effect of Tregs on diabetic cardiomyopathy (DCM) using a db/db mouse model. Eight-week-old male db/db mice were randomly divided into four groups: the control group, administered 200 μL phosphate-buffered saline; the small-dose Treg group, administered 105 Tregs; the large-dose Treg group, administered 106 Tregs; and the PC group, administered 100 μg anti-CD25 specific antibody (PC61) and 106 Tregs. After 12 weeks, mice were euthanized. Transthoracic echocardiography was carried out at the beginning and end of the experiment. Relevant basic experiments to evaluate the effects of Tregs on DCM were carried out. Echocardiography showed that the impaired diastolic and systolic functions were significantly improved in mice administered large-dose Tregs. Large-dose Tregs significantly ameliorated myocardial hypertrophy and fibrosis, and decreased hypertrophic gene expression and collagen deposition. The protective effects of Tregs on diabetic hearts were associated with decreased oxidative stress, inflammatory response and apoptosis. In addition, Tregs promoted the activation of the phosphatidylinositol 3-kinase-protein kinase B signaling pathway, whereas they inhibited extracellular signal-regulated kinase 1/2 and Jun N-terminal kinase phosphorylation, which might be responsible for the cardioprotective role of Tregs against DCM. Tregs ameliorated myocardial hypertrophy and fibrosis, improved cardiac dysfunction, and protected against DCM progression in db/db mice. The mechanisms involved a decrease of inflammatory response, oxidative stress and apoptosis, which might be mediated by phosphatidylinositol 3-kinase-protein kinase B and mitogen-activated protein kinase pathways. Hence, Tregs might serve as a promising therapeutic approach for DCM treatment.

Animal and organoid models to elucidate the anti-fibrotic effect of steroid on biliary atresia

PurposeWe performed animal and organoid study to evaluate the anti-fibrotic effect of steroid on biliary atresia (BA) and the underlying patho-mechanism.MethodsBA animal models were created by inoculation of mice on post-natal day 1 with rhesus rotavirus (RRV). They received either 20 µl phosphate-buffered saline (PBS) or steroid from day 21 to day 34. On day 34, their serum samples were collected for hormonal markers. Necrosis, fibrosis and CK 19 expression in the liver were evaluated. Liver organoids were developed and their morphology as well as bulk RNA sequencing data were analyzed.ResultsTwenty-four mice developed BA features after RRV injection and were equally divided into steroid and PBS groups. On day 34, the weight gain of steroid group increased significantly than PBS group (p < 0.0001). All mice in the PBS group developed liver fibrosis but only one mouse in the steroid group did. Serum bilirubin and liver parenchymal enzymes were significantly lower in steroid group. The morphology of liver organoids were different between the two groups. A total of 6359 differentially expressed genes were found between steroid group and PBS group.ConclusionBased on our findings obtained from RRV-induced BA animal and organoid models, steroid has the potential to mitigate liver fibrosis in BA.

A novel colorimetric strategy based on canine IgM for ultrasensitive and selective detection of Staphylococcus aureus

Conventional methods for Staphylococcus aureus (S. aureus) detection using mammalian immunoglobulins could yield false positive results, owing to the presence of the protein G-producing Streptococcus. In this study, because canine IgM can bind to protein A but not to protein G, we firstly designed a colorimetric platform based on canine IgM to identify S. aureus. Magnetic beads (MBs) functionalized with canine IgM acted as captors to enrich S. aureus, relying on the binding interaction between the Fc region of canine IgM and protein A in the bacterial cell wall, while horseradish peroxidase (HRP)-labeled rabbit anti-S. aureus IgG served as a tracer in the sandwich-type immunoassay, catalyzing the colorimetric reaction. Under optimal conditions, the proposed strategy achieved a satisfactory analytical performance with a broad linear range from 1.6 × 103 CFU/mL to 1.0 × 105 CFU/mL, a colorimetric minimum resolution of as low as 1.4 × 102 CFU/mL in 100 μL phosphate-buffered saline (PBS), and good selectivity in differentiating target from protein G-producing Streptococcus. Moreover, the proposed approach successfully identified S. aureus in various application scenarios (except for samples containing IgG), can be completed in less than 90 min, and was not dependent on any specialized equipment.

Comparison of microscopic and xpert MTB diagnoses of presumptive mycobacteria tuberculosis infection: retrospective analysis of routine diagnosis at Cape Coast Teaching Hospital

IntroductionTuberculosis is a global health problem that causes 1. 4 million deaths every year. It has been estimated that sputum smear-negative diagnosis but culture-positive pulmonary TB diagnosis contribute to 12.6% of pulmonary TB transmission. TB diagnosis by smear microscopy smear has a minimum detection limit (LOD) of 5,000 to 10,000 bacilli per milliliter (CFU/ml) of sputum result in missed cases and false positives. However, GeneXpert technology, with a LOD of 131–250 CFU/ml in sputum samples and its implementation is believe to facilitate early detection TB and drug-resistant TB case. Since 2013, Ghana health Service (GHS) introduce GeneXpert MTB/RIF diagnostic in all regional hospitals in Ghana, however no assessment of performance between microscopy and GeneXpert TB diagnosis cross the health facilities has been reported. The study compared the results of routine diagnoses of TB by microscopy and Xpert MTB from 2016 to 2020 at the Cape Coast Teaching Hospital (CCTH).MethodsThe study compared routine microscopic and GeneXpert TB diagnosis results at the Cape Coast Teaching Hospital (CCTH) from 2016 to 2020 retrospectively. Briefly, sputum specimens were collected into 20 mL sterile screw-capped containers for each case of suspected TB infection and processed within 24 h. The samples were decontaminated using the NALC-NaOH method with the final NaOH concentration of 1%. The supernatants were discarded after the centrifuge and the remaining pellets dissolved in 1–1.5 ml of phosphate buffer saline (PBS) and used for diagnosis. A fixed smears were Ziehl-Neelsen acid-fast stain and observed under microscope and the remainings were used for GeneXpert MTB/RIF diagnosis. The data were analyze using GraphPad Prism.Results50.11% (48.48–51.38%) were females with an odd ratio (95% CI) of 1.004 (0.944–1.069) more likely to report to the TB clinic for suspected TB diagnosis. The smear-positive cases for the first sputum were 6.6% (5.98–7.25%), and the second sputum was 6.07% (5.45–6.73%). The Xpert MTB-RIF diagnosis detected 2.93% (10/341) (1.42–5.33%) in the first and 5.44% (16/294) (3.14–8.69%) in the second smear-negative TB samples. The prevalence of Xpert MTB-RIF across smear positive showed that males had 56.87% (178/313) and 56.15% (137/244) and females had 43.13% (135/313) and 43.85% (107/244) for the first and second sputum. Also, false negative smears were 0.18% (10/5607) for smear 1 and 0.31% (16/5126) for smear 2.ConclusionIn conclusion, the study highlights the higher sensitivity of the GeneXpert assay compared to traditional smear microscopy for detecting MTB. The GeneXpert assay identified 10 and 16 positive MTB from smear 1 and smear 2 samples which were microscopic negative.

EFFECTIVENESS OF CAPPA-CARRAGEENAN TO INCREASE TO RESISTANCE OF FRESHWATER CRAYFISH Cherax quadricarinatus TO Aeromonas hydrophila INFECTION

Crayfish Cherax quadricarinatus is one of the fishery commodities that has business prospects not only as an ornamental fish commodity but also as a consumption commodity. Disease attacks caused by bacteria cannot be avoided, so one of the efforts made to increase the resistance of crayfish by giving kappa carrageenan is to increase the resistance of the crayfish. The purpose of this study was to determine the level of resistance of C. quadricarinatus to A. hydrophila bacterial infection by giving kappa carrageenan with different frequencies which is effective for increasing lobster survival after challenge tests with A. hydrophila bacteria. The study was conducted using a completely randomized design (RAL) with five treatments and three replications, namely control (without injection of kappa carrageenan), 1 time, 2 times, 4 times, and 6 times of injection kappa carrageenan. During maintenance, kappa carrageenan was given at a dose of 5 μg/g of oyster by dissolving 2.5 mg in 1 ml of Phosphate Buffer Saline (PBS) and then injected into lobster as much as 20μl/head. on day 30, challenged with A. hydrophila bacteria. The results showed that giving kappa-carrageenan 2, 4, and 6 times showed the same results (P&gt;0.05) on the absolute growth of crayfish. The survival rate of crayfish given kappa-carrageenan showed that the frequency of giving 4 times showed the highest survival rate. Water quality during the study consisting of oxygen 6.25 mg/l, carbodioxide 3.68 mg/l, pH 6.8, ammonia 0.17 mg/l, and temperature 26.9 °C showed that it was still in the range that was feasible as a maintenance media

MicroRNA-7 attenuates secondary brain injury following experimental intracerebral hemorrhage via inhibition of NLRP3

Background and purposeThe pathophysiological mechanisms underlying brain injury resulting from intracerebral hemorrhage (ICH) remain incompletely elucidated, and efficacious therapeutic interventions to enhance the prognosis of ICH patients are currently lacking. Previous research indicates that MicroRNA-7 (miR-7) can suppress the expression of Nod-like receptor protein 3 (NLRP3), thereby modulating neuroinflammation in Parkinson's disease pathogenesis. However, the potential regulatory effects miR-7 on NLRP3 inflammasome after ICH are yet to be established. This study aims to ascertain whether miR-7 mitigates secondary brain injury following experimental ICH by inhibiting NLRP3 and to investigate the underlying mechanisms. MethodsAn ICH model was established by stereotaxically injecting 100 μL of autologous blood into the right basal ganglia of Sprague-Dawley (SD) rats. Subsequently, these rats were allocated into three groups: sham, ICH + Vehicle, and ICH + miR-7, each comprising 18 animals. Twelve hours post-modeling, rats received intraventricular injections of 10 μL physiological saline, 10 μL phosphate, and 10 μL phosphate-buffered saline solution containing 0.5 nmol of miR-7 mimics, respectively. Neurological function was assessed on day three post-modeling, followed by euthanasia for brain tissue collection. Brain water content was determined using the dry-wet weight method. The expression of inflammatory cytokines in cerebral tissues surrounding the hematoma was analyzed through immunohistochemistry and Western blot assays. These cytokines were re-evaluated using Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Moreover, bioinformatics tools were employed to predict miR-7′s binding to NLRP3. A wild-type luciferase reporter gene vector and a corresponding mutant vector were constructed, followed by transfection of miR-7 mimics into HEK293T cells to assess luciferase activity. ResultsOur study demonstrates that the administration of miR-7 mimics markedly reduced neurological function scores and attenuated brain edema in rats following ICH. A significant upregulation of NLRP3 expression in microglia/macrophage adjacent to the hematoma was observed, substantially reduced after the treatment with miR-7 mimics. Furthermore, this intervention ameliorated neurodegenerative changes and effectively decreased the protein and mRNA levels of pro-inflammatory cytokines, namely TNF-α, IL-1β, IL-6, and Caspase1, in the cerebral tissues proximate to the hematomas. In addition, miR-7 mimics distinctly inhibited the luciferase activity associated with the wild-type reporter gene, an effect not mirrored in its mutant variant. ConclusionsThe miR-7 suppressed NLRP3 expression in microglia/macrophage to reduce the production of inflammatory cytokines, leading to conducting certain neuroprotection post-ICH in rats.

The GLP-1 receptor agonist exenatide improves recovery from spinal cord injury by inducing macrophage polarization toward the M2 phenotype.

Although a wide variety of mechanisms take part in the secondary injury phase of spinal cord injury (SCI), inflammation is the most important factor implicated in the sequelae after SCI. Being central to the inflammation reaction, macrophages and their polarization are a topic that has garnered wide interest in the studies of SCI secondary injury. The glucagon-like peptide 1 (GLP-1) receptor agonist exenatide has been shown to enhance the endoplasmic reticulum stress response and improve motor function recovery after spinal cord injury (SCI). Since exenatide has also been reported to induce the production of M2 cells in models of cerebral infarction and neurodegenerative diseases, this study was conducted to examine the effects of exenatide administration on the inflammation process that ensues after spinal cord injury. In a rat contusion model of spinal cord injury, the exenatide group received a subcutaneous injection of 10 μg exenatide immediately after injury while those in the control group received 1 mL of phosphate-buffered saline. Quantitative RT-PCR and immunohistochemical staining were used to evaluate the effects of exenatide administration on the macrophages infiltrating the injured spinal cord, especially with regard to macrophage M1 and M2 profiles. The changes in hind limb motor function were assessed based on Basso, Beattie, Bresnahan locomotor rating scale (BBB scale) scores. The improvement in BBB scale scores was significantly higher in the exenatide group from day 7 after injury and onwards. Quantitative RT-PCR revealed an increase in the expression of M2 markers and anti-inflammatory interleukins in the exenatide group that was accompanied by a decrease in the expression of M1 markers and inflammatory cytokines. Immunohistochemical staining showed no significant difference in M1 macrophage numbers between the two groups, but a significantly higher number of M2 macrophages was observed in the exenatide group on day 3 after injury. Our findings suggest that exenatide administration promoted the number of M2-phenotype macrophages after SCI, which may have led to the observed improvement in hind limb motor function in a rat model of SCI.

Kelvin probe force microscopy on patterned large-area biofunctionalized surfaces: a reliable ultrasensitive platform for biomarker detection.

Kelvin probe force microscopy (KPFM) allows the detection of single binding events between immunoglobulins (IgM, IgG) and their cognate antibodies (anti-IgM, anti-IgG). Here an insight into the reliability and robustness of the methodology is provided. Our method is based on imaging the surface potential shift occurring on a dense layer of ∼5 × 107 antibodies physisorbed on a 50 μm × 90 μm area when assayed with increasing concentrations of antigens in phosphate buffer saline (PBS) standard solutions, in air and at a fixed scanning location. A comprehensive investigation of the influence of the main experimental parameters that may interfere with the outcomes of KPFM immune-assay is provided, showing the robustness and reliability of our approach. The data are supported also by a thorough polarization modulation infrared reflection-absorption spectroscopy (PM-IRRAS) analysis of the physisorbed biolayer, in the spectral region of the amide I, amide II and amide A bands. Our findings demonstrate that a 10 min incubation in 500 μL PBS encompassing ≈ 30 antigens (100 zM) triggers an extended surface potential shift that involves the whole investigated area. Such a shift quickly saturates at increasing ligand concentration, showing that the developed sensing platform works as an OFF/ON detector, capable of assessing the presence of a few specific biomarkers in a given assay volume. The reliability of the developed methodology KPFM is an important asset in single molecule detections at a wide electrode interface.

Subcutaneous lysophosphatidylcholine administration promotes a febrile and immune response in Holstein heifer calves

Lysophosphatidylcholine (LPC) is immunomodulatory in nonruminants; however, the actions of LPC on immunity in cattle are undefined. Our objective was to study the effects of LPC administration on measures of immunity, liver health, and growth in calves. Healthy Holstein heifer calves (n = 46; age 7 ± 3 d) were randomly assigned to 1 of 4 treatments (n = 10 to 11 calves/treatment): a milk replacer diet unsupplemented with lecithin in the absence (CON) or presence of subcutaneously (s.c.) administered mixed (mLPC; 69% LPC-16:0, 25% LPC-18:0, 6% other) or pure LPC (pLPC; 99% LPC-18:0), or a milk replacer diet supplemented with 3% lecithin enriched in lysophospholipids containing LPC in the absence of s.c.-administered LPC (LYSO) for 5 wk. Calves received 5 s.c. injections of vehicle (10 mL of phosphate-buffered saline containing 20 mg of bovine serum albumin/mL; CON and LYSO) or vehicle containing mLPC or pLPC to provide 10 mg of total LPC per kilogram of BW per injection every 12 h during wk 2 of life. Calves were fed a milk replacer containing 27% crude protein and 24% fat at 1.75% of BW per day (dry matter basis) until wk 6 of life (start of weaning). Starter grain and water were provided ad libitum. Body measurements were recorded weekly, and clinical observations were recorded daily. Blood samples were collected weekly before morning feeding and at 0, 5, and 10 h, relative to the final s.c. injection of vehicle or LPC. Data were analyzed using a mixed model, with repeated measures including fixed effects of treatment, time, and their interaction. Dunnett's test was used to compare treatments to CON. Peak rectal temperatures were higher in mLPC or pLPC, relative to CON. Plasma LPC concentrations were greater in mLPC and LYSO calves 5 h and 10 h after the final injection, relative to CON. Calves receiving mLPC and pLPC also had higher circulating serum amyloid A concentrations, relative to CON. Calves receiving mLPC had greater serum aspartate aminotransferase, γ-glutamyltransferase, and glutamate dehydrogenase concentrations, relative to CON. Calves provided mLPC experienced lower average daily gain (ADG) after weaning, relative to CON. The LYSO treatment did not modify rectal temperatures, ADG, or measures of liver health, relative to CON. We conclude that LPC administered as s.c. injections induced an acute febrile response, modified measures of liver and immune function, and impaired growth in calves.

Molecular detection and pathological modifications of Klebsiella pneumoniae in trachea and lung of rabbits after infected by intranasal instillation route

Klebsiella pneumoniae, one of the more critical zoonotic pathogenic bacteria, causes many diseases, including pneumonia, urinary tract infections, bloodstream ‎infections, and sepsis. This study aimed to insulate K. pneumoniae from sheep ‎urine and identify it by microbiological method on MacConkey agar and confirmed with ‎‎16S-rRNA sequencing analysis, then study the pathological changes ‎of bacteria in the trachea and lung of rabbits after infected by intranasal instillation ‎route. A total of 24 adult healthy local male rabbits were divided randomly into 2 groups: in the control group, eight animals received 50 µL phosphate buffer saline via intranasal instillation. In contrast, sixteen animals received intranasally a single dose of K. pneumonia at 106 CFU/50 µL PBS/animal in the infected group. Two animals from the infected group and one from the control group were sacrificed at ‎‎3 hours, 6 hours, 12 hours, 24 hours, 3 days, 7 days, and 16 days post-infection. A postmortem examination was performed, and any gross lesions were reported. Trachea and lung specimens were then collected and fixed in 10% formalin. The results of the bacterial ‎examination growing on the MacConkey media agreed with the results of molecular ‎confirmed by 16S rRNA sequencing analysis. Macroscopic results highlighted severe lung congestion and white spot areas in the lung tissue. Microscopic ‎changes in the trachea are characterized by acute tracheitis with infiltration of inflammatory cells ‎within a fibro-cartilaginous layer, with sticky exudate consisting of fluid collection and ‎inflammatory cells in the tracheal lumen. as well as bleeding and aggregation edematous ‎fluid recorded in alveolar lumen, with alveolar epithelial sloughing, interstitial pneumonia, ‎bronchiectasis, and bronchiolitis with emphysema. The conclusion of this study could be ‎summarized by addressing the severity of infection at several different times from the Intranasal ‎instillation route with the rapid and dangerous ascending progression of pathological lesions ‎in tracheal and lung disease post-infected rabbits via Intranasal instillation‎. Keywords: Klebsiella pneumoniae,16S-rRNA , trachea, lung, rabbit

Bovine Viral Diarrhea Virus Antibody Level Variation in Newborn Calves after Vaccination of Late-Gestational Cows.

This study was conducted to confirm variation in bovine viral diarrhea virus (BVDV) antibody levels transferred to calves from their mother's colostrum after vaccination of late-gestational cows. Blood samples were drawn from 60 pregnant cows that had been vaccinated more than one year and less than two years previously. The samples were collected six weeks prior to the expected date of delivery. After sample collection, the cows were divided into two groups of 30. One group received 2 mL of BVDV vaccine, and a control group received 2 mL of phosphate-buffered saline (PBS). Blood was collected from the cows three weeks post-administration. Additional blood samples were taken from calves at 1, 4, 8, 12, 16, and 20 weeks after birth. The serum was separated from the collected blood, and BVDV antibody changes were confirmed by enzyme-linked immunosorbent assays. BVDV antibody levels were higher from 8 to 20 weeks of age in calves born to late-gestational BVDV-vaccinated cows than in calves born to control cows (p < 0.0083). Further analysis confirmed a slow decline in BVDV maternal antibodies in calves born to pregnant cows that produced high levels of BVDV antibodies following pre-calving BVDV vaccination. These results suggest that BVDV vaccination of cattle in late pregnancy may help to extend the duration of protection against BVDV infection in newborn calves.

The effects of timing of high immune response phenotyping in relation to weaning on immune responses of crossbred beef calves.

Genetic selection for immune response has the potential to increase the sustainability of the beef industry by breeding cattle that are productive yet with an increased capacity to resist disease. Determining the optimal time to immunophenotype beef cattle is crucial for the accurate prediction of an animal's immune response. The objective of this study was to determine the effect of time of immunophenotyping in relation to weaning on immune responses of beef calves. Antibody- (AMIR) and cell-mediated (CMIR) immune responses were measured on 97 calves on the day of weaning (WEANING, N = 56) or 2 mo post-weaning (POST-WEANING, N = 41). Within each period of immunophenotyping, on day 0, blood was collected, and calves received a 1.0 mL intramuscular injection of type 1 and 2 test antigens. On day 14, blood was collected, and baseline skinfold thickness (SFT) was measured. Calves received an intradermal injection of 0.1 mg of the type 1 antigen suspended in 0.1 mL phosphate buffered saline (PBS) in the right tail fold, and 0.1 mL of PBS in the left. Changes in SFT at 24 h was used to indicate CMIR. To assess AMIR, the titer of type 2 antigen-specific bovine immunoglobulin G in serum from blood collected on day 14 was determined by measuring optical density (OD) using an enzyme-linked immunosorbent assay (ELISA). Among heifers, AMIR was greater for the POST-WEANING group than for the WEANING group (P < 0.01). Among steers, AMIR was not different between the POST-WEANING group and the WEANING group (P = 1.0). Therefore, the AMIR of heifers may be more negatively affected by immunophenotyping at weaning than the AMIR of steers. For steers, CMIR was greater in the POST-WEANING group than the WEANING group (P < 0.001). For heifers, CMIR was not different between the POST-WEANING group and the WEANING group (P = 0.22). The CMIR of steers may be more negatively affected by immunophenotyping at weaning than the CMIR of heifers. Calf age was not associated with AMIR or CMIR for calves phenotyped at weaning or post-weaning. The effect of sire nested within dam age was significant for CMIR for calves in the POST-WEANING group (P < 0.01), but not for calves in the WEANING group (P = 0.67). The results suggest that measuring immunocompetence at weaning may not be representative of a calf's genetic ability to mount an effective immune response, and immunophenotyping should be performed outside the weaning period.

Understanding the role of zingerone on biochemical and behavioral changes in rat brain inflicted with C6 glioma cells.

Malignant glioma is the deadliest form of brain cancer. Zingerone (ZO), a polyphenolic compound found in ginger, offers pharmacological properties that make it a promising agent for containing the growth of glioma cells. The present study was conducted to understand the efficacy of ZO in containing the growth of C6 glioma cells. The study also assessed the prophylactic role of ZO on rat brain glioma induced by C6 cell lines by addressing its antioxidative action on biochemical, behavioral, and histoarchitectural indices. For dose optimization, the animals were pretreated with different doses of ZO for a period of 2 weeks before the inoculation of glioma cells (1 × 105 /10 µL phosphate-buffered saline) in the caudate region of rat brain and the treatment with ZO continued for 4 more weeks post implantation. In vitro studies were done to assess the radical scavenging activity of ZO and also to determine its effects on viability of C6 glioma cells at different concentrations. Glioma-bearing rats showed significant alterations in memory; exploratory and muscular activities which were appreciably improved upon simultaneous supplementation of ZOadministered at a dose of 50 mg/kg body weight and were also visible even at a higher dose. Glioma-bearing rats revealed a significant increase in reactive oxygen species, protein carbonyl contents, and lipid peroxidation, but showed a significant decrease in reduced glutathione and antioxidative enzymes in the brain tissue. Interestingly, all the biochemical indices and altered brain histoarchitecture displaying cellular atypia and hyperplasia showed appreciable improvement when supplemented with ZO at a dose of 50 mg/kg body weight.

Volumetric changes in root canal sealers in ex vivo and a novel animal model approach.

This study aimed to evaluate the volumetric change of root canal sealers through micro-computed tomographic analysis using a novel in vivo model and to compare the results with those obtained using an ex vivo test. Eighteen single-rooted teeth were cut to 5 mm length from the root apex. The root canals were uniformly enlarged and filled with EndoSequence BC Sealer or AH Plus Jet root canal sealers. Samples were stored at 37°C and 95% relative humidity for 24 h and then scanned with a micro-CT device. Twelve samples (n = 6 for each sealer) were implanted in the subcutaneous tissue of Wistar rats, while six samples (n = 3 for each sealer) were immersed in 20 mL of phosphate-buffered saline (PBS) at 37°C at neutral pH. After 7 and 30 days, teeth were removed from subcutaneous tissue or PBS and rescanned. Statistical analysis of volume changes was performed using Shapiro-Wilk's test and independent t-test (p < .05). AH Plus Jet had smaller volume changes (-2.2 to +0.77%) than EndoSequence BC Sealer (-2.0 to +4.0%) (p < .05), in the two tested models. The volume of the root canal sealers decreased over time (p < .05), in vivo. AH Plus Jet results varied between the in vivo and ex vivo results (p < .05), while EndoSequence BC Sealer presented similar volume losses for both experimental models (p > .05). EndoSequence BC Sealer lost more volume than AH Plus Jet. The experimental conditions influenced the volumetric change of AH Plus Jet but not the EndoSequence BC Sealer. The ex vivo model should be further explored as a methodological alternative to assess the volumetric changes of root canal sealers without causing harm to animals.

Propolis Silver Nanoparticles as an Adjuvant in Immunization of Rats with Citrobactor Freundii Antigens.

This study aimed to examine the impact of Citrobacter freundii killed whole cell sonicated antigen (KWCSAg) alone and in combination with propolis nanoparticles on humoral immunoglobulin (IgG) and cellular immune responses of rats. The ELISA interleukin 4 (IL4) and IgG, delayed-type hypersensitivity (DTH) skin test, and phagocytosis activity tests were used in this study. In total, 45 rats were divided into five groups of 9 rats. The first group received a 1,000 μg\ml dose of KWCSAg-CF. The second group received an injection of 1,000 μg/ml of KWCSAg-CF antigen and 30 mg/ml of propolis AgNPs. The third group received an injection of 1,000 μg/ml of KWCSAg-CF antigen along with 10 mg/ml of propolis AgNPs. The fourth group was subjected to 30 mg/ml of propolis AgNPs. One ml of phosphate-buffered saline (pH 7.2) was injected into the fifth group (the negative control group). The rats received booster injections of the same antigens after 14 days. Blood was obtained from them to detect immunoglobulin and interleukin 4 (IL-4) on days 21, 28, 32, 46, 50, and 60 following the injection. The second group showed the most significant rise in IL-4 and IgG concentration, followed by the third group, the first group, and the fourth group, compared to the negative control group (fifth group). In all immunized groups, the DTH test results demonstrated an increase in the means of induration with significant differences (P<0.05) of the concentrated antigen after 24 and 48 h, and subsequently a decrease after 72 h, compared to the negative control group. At 48 h after the concentrated antigen was indurated, the second group displayed the most significant increase in diameter.

Red osier dogwood extract versus Trimethoprim-sulfadiazine (Part 1). Effects on the growth performance, blood parameters, gut histomorphometry, and Salmonella excretion of broiler chickens orally challenged with Salmonella Enteritidis

The poultry industry has not been spared from the prevalent incidence of diseases caused by invasive pathogens, especially Salmonella. Due to the pressing need to identify a suitable antibiotic alternative for use in poultry production, this study investigated the efficacy of red osier dogwood (ROD) extract on the growth, blood parameters, gut morphology, and Salmonella excretion in broiler chickens orally challenged with Salmonella Enteritidis (SE). A 4×2 factorial experiment was conducted based on 2 main factors, namely dietary treatments, and SE challenge. A total of 404, one-day-old male Ross broiler chicks were randomly assigned to 4 dietary treatments; 1) Negative control (NC), 2) NC+0.075 ppm of Trimethoprim-sulfadiazine (TMP/SDZ)/kg of diet, 3) NC+0.3% ROD extract, and 4) NC+0.5% ROD extract. The absence of SE in the fecal samples obtained from chick delivery boxes was confirmed on d 0. On d 1, half of the birds were orally gavaged with 0.5 mL of phosphate-buffered saline each (noninfected group) and the remaining with 0.5 mL of 3.1×105 CFU/mL SE (infected group) in all treatment groups. Dietary treatments were randomly assigned to 8 replicate cages at 6 birds/cage. On 1-, 5-, 12-, and 18-day postinfection (DPI), cloacal fecal samples were collected on the 6 birds/cage to assess SE excretion. Average weight gain (AWG), average feed intake (AFI), feed conversion ratio (FCR), and mortality were determined weekly. On d 21, 10 chickens/treatment were euthanized to perform hematology, gut histomorphometry, serum immunoglobulins G and M (IgG and IgM), and superoxide dismutase measurements. Both ROD extract levels did not affect (P > 0.05) growth performance; however, the SE-infected birds showed increased (P < 0.05) AFI and FCR throughout the experimental period. Regardless of the SE-infection, both ROD extract levels improved (P < 0.05) duodenal villus height: crypt depth compared to other treatments. 0.5% ROD extract improved (P < 0.05) ileal villus width (VW) of noninfected birds and ileal crypt depth of infected birds, but it decreased (P < 0.05) the ileal VW of infected birds, compared to other treatments. The SE-infected birds showed lower (P < 0.05) lymphocytes (L) but increased (P < 0.05) heterophils (H), H:L, and monocytes (MON). Both ROD extract levels did not affect (P > 0.05) white blood cell differential, while dietary 0.3% ROD extract increased (P < 0.05) MON of the birds, regardless of infection model. Regardless of infection model, both TMP/SDZ and 0.5% ROD extract reduced the concentration of IgM in the serum, compared to the control and 0.3% ROD (P=0.006). Conclusively, both ROD extract levels improved duodenal histomorphology and body defense against SE infection in broiler chickens; however, the 0.3% ROD extract was better.

Effects of milk-derived bioactive peptide VPP on diarrhea of pre-weaning calves.

A well-known milk-derived bioactive tripeptide, VPP (Val-Pro-Pro) has good anti-inflammatory, anti-hypertension, and anti-hydrolysis properties. However, whether VPP can alleviate calf intestinal inflammation is unclear. In this experiment, the effects of VPP on growth, diarrhea incidence, serum biochemical indices, short-chain fatty acids, and fecal microorganisms were examined in pre-weaning Holstein calves. Eighteen calves with similar birth date, body weight, and genetic background were randomly assigned equally to two groups (n = 9). The control group was given 50 mL of phosphate buffer saline before morning feeding, whereas the VPP group received 50 mL of VPP solution (100 mg/kg body weight/d). The study lasted for 17 days, with the first 3 days used for adaptation. Initial and final body weights were determined, and daily dry matter intake and fecal score were recorded throughout the study. Serum hormone levels and antioxidant and immune indices were measured on day 14. Fecal microorganisms were collected on days 0, 7, and 14, and 16S rDNA sequencing was performed. Oral administration of VPP did not significantly affect calf average daily feed intake and body weight, but the growth rate in body weight was significantly higher in the VPP group than in the control group on day 7 (P < 0.05). Compared with the control, VPP significantly decreased serum TNF-α and IL-6 contents (P < 0.05), and concentrations of nitric oxide and IL-1β also decreased but not significantly (0.05 < P < 0.1). After seven days of VPP, relative abundances of g_Lachnoclostridium, uncultured_bacterium_, and g_Streptococcus in fecal samples increased significantly (P < 0.05). Compared with the control, VPP significantly increased concentrations of the fecal short-chain fatty acids n-butyric acid and isovaleric acid (P < 0.05). In conclusion, VPP can relieve intestinal inflammation and alleviate the degree of diarrhea in pre-weaning calves.

Bead size has a greater effect on in vitro elution from antimicrobial-impregnated calcium sulfate beads than drug concentration

To compare the elution characteristics of amikacin-impregnated calcium sulfate (CaSO4) beads based on different drug concentrations and bead size configurations. Six groups of amikacin-impregnated CaSO4 beads and one negative control group. Amikacin-impregnated CaSO4 beads were formed with either 500 mg (low-concentration) or 1 g (high-concentration) of amikacin per 15 g CaSO4 hemihydrate powder. The number of beads necessary to approximate 150 mg of amikacin for each of the 3 bead sizes (3 mm, 5 mm, and 7 mm) at both low and high concentrations were placed in 6 mL of phosphate-buffered saline. The saline was sampled at 14 time points over 28 days. Amikacin concentrations were determined using liquid chromatography-mass spectrometry. Smaller beads reached higher mean peak concentrations than larger beads (P < .0006). Peak concentrations for the low- and high-concentration groups were 20.5 mg/mL and 27.4 mg/mL, 13.1 mg/mL and 14.0 mg/mL, and 8.85 mg/mL and 6.75 mg/mL for the 3 mm, 5 mm, and 7 mm beads, respectively. Bead size also affected the length of therapeutic duration, lasting 6 days for the 3 mm and 5 mm beads and 9 days for the 7 mm beads. However, this was only statistically evident among the high-concentration beads (P < .044). Antimicrobial concentration within the same bead sizes did not affect elution. Amikacin-impregnated CaSO4 beads achieved extreme supratherapeutic eluent concentrations. While additional studies are needed, bead size significantly affected elution with smaller beads reaching higher peak concentrations and 7 mm, high-concentration beads demonstrating a longer therapeutic duration than smaller beads.

Immune response study of salmonella mbandaka isolated from human in (balb/c) mice

The bases of this study is to evaluate the immune response to experimental infection of (BALB/c) mice by Salmonella Mbandaka . The experiment was carried out on one hundred mice of both genders with age range (6 – 8) weeks old, the mice were divided randomly into two groups (group A: includes 50 mice were given orally on infective dose (1.3×107C.F.U\ml) ,group B: includes 50 mice which inoculated orally with 1 ml of phosphate buffer saline (pH=7.2) and considers as control group).The study has showed that the experimentally infected mice (group A) are able to induce humoral and cellular immune response . The humoral immune response was detected by Enzyme-linked Immunosorbent Assay post induced infection in mice with infected dose. Maximum titers of ( IgG ) antibodies reached of (211.13) at four weeks , slight reduction at six weeks , reached a mean (179.41 ± 29.47), while in the eight weeks after infection, there was a reduction in the(IgG) titers to reach a mean (58.83 ± 34.56). While group B showed the normal range of IgG titer (20.22 ± 1.11 , 20.79 ±1.71 , 19.81 ± 0.88, 20.22 ± 1.11 , 20.97 ± 0.57 ) at zero time , 2 ,4 , 6 ,8 weeks respectively . The cellular immune response was detected by using the skin test mean thickness of the right foot pad was (2.85±0.06) millimeter after 24 hours, then increased to reach (3.69 ±0.07) mm after 48 hours , then declined to reach (2.71±0.09)mm after 72 hours in mice infected by infective dose. This study concluded that Salmonella Mbandaka infection of the host was able to induce both humoral and cellular immune responses, and these responses are dose dependent.

Red osier dogwood extract vs. trimethoprim-sulfadiazine (Part 2). Pharmacodynamic effects on ileal and cecal microbiota of broiler chickens challenged orally with Salmonella Enteritidis

With the subsisting restrictions on the use of antibiotics in poultry production, the use of plant extracts has shown some promising antimicrobial capacity similar to antibiotics; however, such capacity is largely dependent on their total polyphenol concentration and profile. Given the emerging antimicrobial potential of red osier dogwood (ROD) extract, the study aimed to investigate the pharmacodynamic effect of ROD extract on the ileal and cecal microbiota of broiler chickens challenged orally with Salmonella Enteritidis (SE). A 21 d 4×2 factorial experiment was conducted based on 2 main factors, including diets and SE challenge. A total of 384 one-day-old mixed-sex Cobb-500 broiler chicks were randomly allotted to 4 dietary treatments; Negative control (NC), NC+0.075 mg trimethoprim-sulfadiazine (TMP/SDZ)/kg of diet, and NC containing either 0.3 or 0.5% ROD extract. On d 1, half of the birds were orally challenged with 0.5 mL of phosphate-buffered saline (Noninfected group) and the remaining half with 0.5 mL of 3.1×105 CFU/mL SE (Infected group). Dietary treatments were randomly assigned to 8 replicate cages at 6 birds/cage. On d 21, 10 birds/treatment were euthanized and eviscerated to collect ileal and cecal digesta for gut microbiota analysis. The ileal and cecal microbiota was dominated by phyla Firmicutes, Proteobacteria, and Actinobacteriota. The SE infection decreased (P < 0.05) the relative abundance of Proteobacteria and Actinobacteriota in the ileum and ceca, respectively, however, it increased (P < 0.05) Proteobacteria in the ceca. Both 0.3 and 0.5% ROD extracts (P < 0.05) depressed the relative abundance of Actinobacteriota in the ileum but marginally improved (P < 0.05) it in the ceca compared to the TMP/SDZ treatment. Dietary TMP/SDZ increased (P < 0.05) genus Bifidobacterium at the ileal and cecal segments compared to other treatments. Dietary 0.3 and 0.5% marginally improved (P < 0.05) Bifidobacterium in the ceca and depressed (P < 0.05) Weissella and was comparably similar to TMP/SDZ in the ileum. Regardless of the dietary treatments and SE infection, alpha diversity differed (P < 0.05) between ileal and cecal microbiota. Beta diversity was distinct (P < 0.05) in both ileal and cecal digesta along the SE infection model. Conclusively, both ROD extract levels yielded a pharmacodynamic effect similar to antibiotics on ileal and cecal microbiota.