Ml Of Phosphate-buffered Saline Research Articles

The impact of combined thymol and rosmarinic acid on the intestinal microbiota and barrier function of the piglets challenged by Escherichia. coli K88

It has been found that thymol (Thy) and rosmarinic acid (Ros-A) improve the growth performance of piglets and relieve intestinal inflammation in animals. The effects of Thy and Ros-A separately or in combination (Thy×Ros-A) on the intestinal function and health of piglets challenged with Escherichia coli K88 (E. coli K88) were investigated. A total of 30 piglets aged 21 d were assigned to 5 groups (n = 6). The control (Con) and K88 groups piglets received a basal diet, while the Thy, Ros-A, and Thy×Ros-A groups were fed a basal diet supplemented with 500 mg/kg Thy, 500 mg/kg Ros-A, and 250 mg/kg Thy + 250 mg/kg Ros-A, respectively. On the 19th and 20th day, piglets in the K88, Thy, Ros-A, and Thy×Ros-A groups were orally administered 10 mL of phosphate-buffered saline (PBS) containing approximately 1 × 109 colony-forming units/mL of E. coli K88, while the Con group received an equal volume of PBS. The results showed that the Thy×Ros-A treatment reduced the damage to ileal villi induced by the E. coli K88 challenge, leading to longer villi in the ileum (P < 0.05). Thy and Ros-A modulated the composition of the ileal microbiota. Compared to the K88 group, the Thy×Ros-A group had a higher abundance of Lactobacillus and Romboutsia, while Escherichia-Shigella and Desulforvibrio were lower (P < 0.05). Additionally, the Thy×Ros-A group showed elevated levels of gene and protein expressions for zonula occludens-1, occludin, and claudin-1 compared to the K88 group (P < 0.05). In conclusion, combining Thy and Ros-A reduced ileal damage and relieved the inflammation in weaned piglets challenged with E. coli K88 by regulating intestinal microflora and improving barrier function.

Extended release of ciprofloxacin from commercial silicone-hydrogel and conventional hydrogel contact lenses containing vitamin E diffusion barriers.

Vitamin E could be used as a coating with commercial silicone hydrogel lenses to extend the release of various ophthalmic drugs. This concept could provide a promising approach to improve overall ocular therapeutic outcomes for topical ocular drugs. This study aimed to develop a contact lens-based ocular drug delivery system using vitamin E as a diffusion barrier to extend the release duration of ciprofloxacin. Five commercial lenses were soaked for 24 hours in various concentrations of vitamin E dissolved in ethanol (0.0125 to 0.2 g/mL). The lenses were loaded with ciprofloxacin for 24 hours in 3 mL of 3 mg/mL of ciprofloxacin/acetic acid solution. The drug release was evaluated in 3 mL of phosphate-buffered saline solution. At t = 0.5, 1, 2, 4, 6, 8, 12, 16, and 24 hours, the amount of ciprofloxacin released was measured using a UV-VIS spectrophotometer at 270 nm. There was a decrease in ciprofloxacin loading with increasing amounts of vitamin E loaded into the silicone hydrogel lenses. For each lens type, there was an optimal amount of vitamin E loaded that extended the release duration of the drug from 1 hour (without vitamin E) to as long as 16 hours. In contrast, vitamin E loaded into hydrogel lenses had no effect on the amounts of drugs loaded or the release duration. Vitamin E can be used as a diffusion barrier with commercially available silicone hydrogel lenses to provide sustained release of ciprofloxacin. The results suggest that vitamin E may form blockages in channels within a silicone hydrogel lens material, thereby forcing a longer path for drugs to diffuse into and out of the lens material. There is an optimal amount of vitamin E that needs to be loaded to extend the release duration, and this is lens material dependent.

The Effect of Exosomes from Adipose derived Mesenchymal Stem Cells in Photoaged Rat Skin versus Topically Applied Growth Factors. A Histological Study

Abstract Introduction Skin photoaging is characterized by coarse wrinkles, loss of elasticity and dryness. The worst scenario of ultraviolet (UV) ray exposure can reach up to skin malignant changes. The topical application of growth factors has proven to be effective in ameliorating the photoaging. Among the paracrine products of stem cells are exosomes which also play an important role. Aim of the Work To investigate the effect of exosomes derived from adipose mesenchymal stem cells on the histological structure of photoaged rat thin skin versus topically applied growth factors. Materials and Methods Forty-five young adult male albino rats aged between 6 - 8 weeks were used in this study. Animals were subjected to gentle shaving of the hair over the center of their backs and they were randomly classified into four groups. Control group (I), 20 rats were divided into four subgroups (five animals each). Subgroup Ia, animals were left without treatment, Subgroup Ib, animals were injected subcutaneously with 1 ml of Phosphate Buffer Saline (PBS) at the shaved areas. Subgroup Ic, shaved areas were topically treated with growth factors gel after micro-needling procedure (Three topical treatments were repeated each one performed every 10 days). Subgroup Id, rats were subcutaneously injected with one dose of purified exosome concentrate containing 300 µg of total protein. Group (II)– (photoaged untreated group), five rats, each was exposed to UVA + UVB irradiation for two weeks (five days per week) on the shaved areas using UV lamp. Group (III)- (photoaged exosome-treated group), five rats, each was exposed to UVA + UVB irradiation similar to group II. The animals were injected with one dose of purified exosome concentrate containing 300 µg of total protein subcutaneously at the shaved areas. Group (IV)- (Photoaged and topically treated with growth factors), five rats each was exposed to UVA + UVB irradiation similar to group II. Then, the animals were treated topically as in subgroup C. The back skin was dissected and processed to prepare sections for light microscopic examination with H&amp;E, Masson’s trichrome, Orcein and PAS stains. Immunostaining for KI67and CD31 were done. Morphometric and statistical analysis were performed. Results Group II revealed significant decrease in the epidermal thickness, hair follicles were scanty and disorganized. Orcein sections showed short fragmented elastic fibers and irregular arrangement of collagen fibers with Masson’s trichrome stain. KI67 positive cells was higher than other groups and CD31 was the opposite. Groups III and IV showed marked decrease in the previously mentioned microscopic changes denoting amelioration of the recorded photoaged epidermal and dermal changes. Conclusion The current study concluded that both exosomes and topically applied Growth factors over three divided doses accompanied with micro-needling have resulted in a significant amelioration of the altered microscopic photoaging changes observed after exposure of the rat thin skin to the UV rays. Furthermore, the improvement was more obvious with the topically applied growth factors compared to one dose of exosomes concentrate. Further studies are recommended using repeated application of exosomes and also in combination with micro- needling to stabilize the best protocol for treatment.

Thymoquinone ameliorate oxidative stress, GABAergic neuronal depletion and memory impairment through Nrf2/ARE signaling pathway in the dentate gyrus following cypermethrin administration

BackgroundExposure to chemical toxins, including insecticides, harms bodily organs like the brain. This study examined the neuroprotective of thymoquinone on the cypermethrin’s harmful effects on the histoarchitecture of the dentate gyrus and motor deficit in the dentate gyrus.MethodsForty adult male rats (180–200 g) were randomly divided into 5 groups (n = 8 per group). Groups I, II, III, IV, and V received oral administration of 0.5 ml of phosphate-buffered saline, cypermethrin (20 mg/kg), thymoquinone (10 mg/kg), cypermethrin (20 mg/kg) + thymoquinone (5 mg/kg), and cypermethrin (20 mg/kg) + thymoquinone (10 mg/kg) for 14 days respectively. The novel object recognition test that assesses intermediate-term memory was done on days 14 and 21 of the experiment. At the end of these treatments, the animals were euthanized and taken for cytoarchitectural (hematoxylin and eosin; Cresyl violet) and immunohistochemical studies (Nuclear factor erythroid 2-related factor 2 (Nrf2), Parvalbumin, and B-cell lymphoma 2 (Bcl2).ResultThe study shows that thymoquinone at 5 and 10 mg/kg improved Novelty preference and discrimination index. Thymoquinone enhanced Nissl body integrity, increased GABBAergic interneuron expression, nuclear factor erythroid 2-derived factor 2, and enhanced Bcl-2 expression in the dentate gyrus. It also improved the concentration of nuclear factor erythroid 2-derived factor 2, increased the activities of superoxide dismutase and glutathione, and decreased the concentration of malondialdehyde level against cypermethrin-induced neurotoxicity.Conclusionthymoquinone could be a therapeutic agent against cypermethrin poisoning.

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.

In vitro elution of amikacin, cefazolin, gentamicin, ampicillin/sulbactam, and meropenem from a commercially available calcium sulfate delivery kit.

The use of implantable antibiotic beads has become a frequent treatment modality for the management of surgical site infections in human and veterinary medicine. The objective of this study is to describe the elution kinetics of five antibiotics from a commercially available calcium sulfate antibiotic delivery kit. A secondary goal was to compare elution concentrations with minimal inhibitory concentrations (MIC) for commonly encountered bacteria from the University of Florida's veterinary microbiology laboratory database. Calcium sulfate powder was combined with amikacin, cefazolin, gentamicin, ampicillin/sulbactam, and meropenem. Triplicates of three antibiotic-loaded beads were immersed in 5 mL of phosphate-buffered saline (PBS) and kept at 37°C under constant agitation. Antibiotic-conditioned PBS was sampled at 14 time points from 1-h to 30 days and analyzed by liquid chromatography to determine the antibiotic concentration. All beads eluted concentrations of antibiotics for the 30-day sampling period, except for ampicillin/sulbactam, with the most antibiotics being eluted within the first week. The concentration of antibiotics within the eluent within the first 3-9 days (3- and 5-mm beads, respectively) was greater than the MIC of common isolates. The 5 mm bead samples were superior in maintaining higher concentrations for a longer period, compared to the 3-mm beads. CSH beads eluted antibiotics over the 30-day course of the study. Most of the antibiotic elution occurred within the first week and was maintained above the MIC of commonly encountered isolates. This information may be useful for clinical decision making for treatment of local infections encountered in practice.

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.