Total Cell Count Research Articles

Increased apoptosis in late-developing in vitro fertilized bovine blastocysts decreases successful pregnancy.

Pregnancy in cattle after embryo transfer (ET) is influenced by several factors, including embryo quality. Therefore, preparing high-quality embryos with the greatest developmental potential is essential for achieving a successful pregnancy after ET. Meanwhile, blastocysts produced by in vitro fertilization (IVF) procedure have different developmental speed during in vitro culture (IVC) and they exhibited different competence in the establishment of pregnancy. This study aimed to identify the comparative features of early-, mid-, and late-developing bovine IVF blastocysts, when they first appeared at Day 7, 8, and 9 during IVC, respectively. In addition, the correlations between their molecular features and pregnancy ability were analyzed. The results showed no difference in the morphological characteristics, including total cell count and diameter, between the Day 7, 8, and 9 blastocysts. However, the pregnancy rate post-ET was significantly different between the groups at 51.7%, 36.7%, and 17.8% for Day 7, 8, and 9 blastocysts, respectively. During early embryo development, late-developing blastocysts demonstrated a reduced cell count in the inner cell mass and decreased expression of the early embryo developmental genes (Oct4 and Sox2) compared with the early- and mid-developing blastocysts. In addition, the number of apoptotic cells and apoptosis-related gene expression (increased Bax and decreased Bcl2) gradually elevated from the Day 7 to Day 9 blastocysts. However, there was no difference in mitochondrial activity and mitochondria-relevant gene expression (Tfam and Cox1) between the groups. Correlation analysis identified a significantly negative correlation between the pregnancy rate and the blastocysts' degree of apoptosis, indicating that the low pregnancy ability of late-developing blastocysts was mainly caused by increased apoptosis. This study's results may contribute to the field of animal biotechnology by assisting in establishing an improved strategy for bovine ET with IVF embryos.

Hydrogen gas inhalation ameliorates glomerular enlargement after hypoxic-ischemic insult in asphyxiated piglet model

Acute kidney injury (AKI) has been reported to occur in 30–70% of asphyxiated neonates. Hydrogen (H2) gas became a major research focus in neonatal medicine after the identification of its robust antioxidative properties. However, the ability of H2 gas to ameliorate AKI is unknown. We examined histopathological injuries in the piglet renal cortex on day 5 after a hypoxic-ischemic (HI) insult and if H2 gas can alleviate kidney injuries. Twenty piglets were divided into three groups: no insult (Control, n = 6), HI insult alone (HI, n = 8), and HI insult with H2 gas ventilation (HI-H2, 2.1–2.7% for 24 h, n = 6). The total glomerular cell count was significantly higher in the HI group than in the other groups, with no difference between the HI-H2 and control groups. Proximal tubular lumen narrowing was significantly increased in the HI group versus control, but not in the HI-H2 group. In this piglet model, glomerular enlargement with an increase in glomerular cell number due to tubular lumen narrowing was observed on day 5 after HI insult. H2 gas effectively suppressed this glomerular cell increase and tubular lumen narrowing.

The Protective Effects of Annexin A1 in Acute Lung Injury Mediated by Nrf2.

Acute lung injury (ALI), one of the most severe respiratory system diseases, is prevalent worldwide. Annexin A1 (AnxA1) is an important member of the annexin superfamily, known for its wide range of physiological functions. However, its potential protective effect against lipopolysaccharide (LPS)-induced ALI remains unclear. Mice were divided into four groups: Sham, LPS + vehicle, LPS + 0.1 μg AnxA1, and LPS + 0.5 μg AnxA1. Lung injury was assessed through histopathology, pulmonary wet-to-dry (W/D) ratio, cell counting of bronchoalveolar lavage fluid (BALF), oxidative stress analysis, and noninvasive pulmonary function testing. Gene and protein expression levels were measured using RT-PCR, ELISA, and western blot analysis. AnxA1 alleviated LPS-induced ALI by protecting lung tissue from damage, reducing the lung wet/dry (W/D) weight ratio, and improving LPS-induced impaired lung function. Interestingly, administration of AnxA1 was found to repress the infiltration of inflammatory cells by decreasing the total cell count, neutrophils, and protein concentrations in bronchoalveolar lavage fluid (BALF). AnxA1 mitigated the inflammatory response in the pulmonary tissue by lowering the levels of IL-1β, IL-6, and TNF-α in BALF of ALI mice. Additionally, AnxA1 attenuated oxidative stress in lung tissues of ALI mice by restoring the activity of catalase (CAT), SOD, and glutathione (GSH) but reducing the levels of malondialdehyde (MDA). We also found that AnxA1 suppressed activation of the NLRP3 signaling pathway. Mechanistically, AnxA1 activated the Nrf2/HO-1 signaling pathway while preventing the activation of NF-κB. Collectively, these findings suggest that AnxA1 alleviates LPS-induced ALI and might be a promising novel therapeutic agent against LPS-induced ALI.

Immunopathological Changes of Streptococcus pneumoniae Causing Respiratory Infection in Lambs

Streptococcus pneumoniae are common bacterial pathogens that can cause a range of diseases in humans and animals. This study attempts to clarify the course and consequences of pneumonia caused by this bacterial infection. A total of 6 male un-weaned lambs aged between 1 to 2 months and weighing 5 to 7 kg were subjected to S. pneumoniae strain ATCC 6303 serotype 3 at ‎2×106 ‎CFU/mL by inhalation to induce pneumonia. Pneumonic clinical signs were ‎monitored daily throughout the study period. The blood samples were collected from all animal groups at day zero before pneumoniae induction and 3, 6, and 14 days post infection for total and differential white blood cell count ‎(WBCs) and tumor necrosis factor-alpha (TNF-α) assessments. Additionally, on days 6 and 14 post-exposure, trachea and lung tissue samples were harvested for macroscopic and microscopic pathological changes evaluation. The results showed that there was a significant increase (P<0.001) in total WBC counts from day 3 post-exposure and maintaining elevated levels on days 6 and 14 compared to day zero. Differential WBC counts showed an early, significant rise in neutrophils, with sustained ‎elevation in lymphocytes and monocytes. TNF-α‎ levels significantly elevated on day 3‎ and gradually declined by day 14‎ post-exposure (P<0.001). On day 6 post-exposure, the gross pathological changes in the lung showed pulmonary edema, and emphysema, with mild to moderate lung congestion and emphysematous changes observed on day 14 post-exposure. Histopathologically, severe erosion and necrosis in the trachea and bronchus epithelium, along with inflammation in adjacent mucosa and submucosa, accompanied by focal inflammatory infiltration and emphysema in the lung by day 6 post-exposure, were observed. By day 14, these changes progressed to marked epithelial vacuolation and necrosis in the trachea, with lung sections revealing perivascular cuffing, peri bronchiolitis, mild bronchiectasis, atelectasis, and alveolar collapse. This study is attempt for a better understanding of S. pneumoniae infection in lamb and contributing to the development of preventive and management strategies in Iraq‎‎‎.

Clinicopathologic Parameters of Peritoneal Fluid as Predictors of Gastrointestinal Lesions, Complications, and Outcomes in Equine Colic Patients: A Retrospective Study.

Neutrophil characteristics in peritoneal fluid (PF) may aid in diagnosing and treating specific colic lesions and complications. The objective of this retrospective study was to evaluate quantitative PF leukocyte values, as well as PF total protein (TP) and lactate, for associations with diagnosis, morbidity, and mortality in horses with acute colic. Three hundred and forty-two horses that presented to one institution between January 2010-2020 for the evaluation of acute colic were included. The PF total nucleated cell count (TNCC), % and total neutrophil counts, total protein (TP), and lactate were analyzed for associations with lesion location and type, the development of postoperative reflux (POR) or systemic inflammatory response syndrome (SIRS), and survival to discharge via Kruskal-Wallis testing. Horses with strangulating lesions had higher PF % neutrophils, neutrophil count, and TNCC compared to non-strangulating lesions. The development of SIRS or POR was associated with higher PF TNCC, total neutrophil count, TP, and lactate. Horses that did not survive to discharge had increased PF % neutrophils, neutrophil count, TP, lactate, and ratio of PF-to-systemic TP than those that survived via univariable analysis. Identified associations between increased PF neutrophils and the development of POR and SIRS warrant further investigation to better understand their role in the pathogenesis of equine colic and potential as targets for therapeutic intervention.

Gastrointestinal Dysfunction Bears on the Clinical-Biological Profile of Parkinson's Disease.

Gastrointestinal dysfunction (GID) accompanies any phase of Parkinson's disease (PD), underlying differential clinical-pathological trajectories. To investigate associations between GID and peripheral immune or neurodegeneration-related markers in PD. One-hundred-and-fourteen patients (n = 55 de novo, DN; n = 59 middle-advanced, MA) completed the Gastrointestinal Dysfunction Scale for PD (GIDS-PD), and other motor and non-motor scales; paired measurement of amyloid-β42, amyloid-β42β/β40, total-tau, phosphorylated-181-tau, total α-synuclein CSF levels, albumin ratio, and peripheral blood cell count were collected. Group and correlation analyses were performed. MA patients had greater GID than DN. GIDS-PD scores directly correlated with MDS-UPDRS-III and non-motor scores in both groups, although more in DN. GIDS-PD scores were directly associated with α-synuclein and inversely with lymphocytes only in DN; conversely, they were positively associated with tau proteins and albumin ratio, and negatively with amyloid-β-peptides in both groups. The burden of GID increases along the PD course with associated stage-specific clinical-biological patterns.

Feasibility and Safety of Autologous Cord Blood Derived Cell Administration in Extremely Preterm Infants: A Single-centre, Open-label, Single-arm, Phase I trial (CORD-SaFe study)

Evidence from preclinical studies in small and large animal models has shown neuroprotective effects of intravenous administration of umbilical cord blood derived cells (UCBCs). This study aimed to evaluate the feasibility of umbilical cord blood (UCB) collection, extraction of UCBCs, and subsequent safety of intravenous autologous administration of UCBCs in extremely preterm infants (born <28 weeks gestation). A single-centre, open-label, single-arm, safety and feasibility clinical intervention trial was conducted at Monash Medical Centre and Monash Children's Hospital, Melbourne, Australia. Participants were extremely preterm infants born at less than 28 weeks completed gestation, and exclusions included major congenital malformation, maternal blood-borne virus infection, and severe brain injury on postnatal cranial ultrasound. UCB was collected at birth, and UCBCs were characterised (total nucleated cell count (TNC), mononuclear cell count (MNC), CD34+ cell count) and cryopreserved. Infants were reinfused with autologous UCBCs (25-50 million MNCs/kg) intravenously in the second postnatal week. Primary outcomes included feasibility: sufficient UCB volume (>7mL) and UCBC numbers following processing (>25×106TNCs/kg); and safety: absence of adverse events directly related to UCBC administration. Forty-four UCB collections were attempted and sufficient UCB volume/UCBC extraction was demonstrated in 37 (84.1%) infants. Good Manufacturing Practice (GMP) grade cells were obtained in 31/44 (70.4%) of infants. Median (IQR) TNCs and MNCs collected were 130 (67-207) x 106/kg and 60 (39-105) x 106/kg, respectively. 23 infants with median (IQR) gestation of 26 (24-27) weeks and birth weight of 761 (650-946) grams were administered cells at a median (IQR) dose of 42.3 (31.1-62.3) x 106 MNCs/kg). No serious adverse events were noted, and the infusions were well-tolerated. This phase-1 clinical trial has shown UCBC collection and reinfusion was feasible in approximately 70% of extremely preterm infants and was well tolerated without any serious adverse events. Funding to support this study was obtained from National Health and Medical Research Council of Australia, Cerebral Palsy Alliance, National Stem Cell Foundation of Australia, and Lions Cord Blood Foundation.

Clinical presentation, diagnostic findings and short-term outcome in French bulldogs with presumptive spinal only meningoencephalomyelitis of unknown origin: 15 cases (2016-2023).

To describe clinical presentation, diagnostic findings and outcome of presumptive spinal only meningoencephalomyelitis of unknown origin in French bulldogs, and to compare clinical presentation between intervertebral disc herniation and spinal only meningoencephalomyelitis of unknown origin in this breed. Medical records of French bulldogs diagnosed with presumptive spinal only meningoencephalomyelitis of unknown origin in seven referral centres were reviewed. Clinical presentation was compared to a group of French bulldogs with intervertebral disc herniation. Fifteen French bulldogs were included. Median age at presentation was 31 months. Most dogs were presented with a chronic onset (54%), and the median duration of clinical signs before diagnosis was 21 days. The most common presentation was ambulatory paresis and proprioceptive ataxia (80%). Spinal pain was uncommon (20%). There was no preferential location between the cervical and thoracolumbar segments. In all but one case, magnetic resonance imaging revealed broad focal or multifocal, poorly defined, intramedullary lesions that typically appeared hyperintense on T2W images and isointense on T1W images with parenchymal and/or meningeal contrast enhancement in 12 dogs (80%). Median total nucleated cell count in cerebrospinal fluid was 15 cells/mm3 with predominance of mononuclear cells in all cases. Follow-up (available for 12 dogs, ranging from 1 month to 1 year) indicated initial positive response to various immunosuppressive treatments in all but one case, with subsequent suspected relapse in three cases. Dogs with spinal only meningoencephalomyelitis of unknown origin had longer clinical sign duration compared to intervertebral disc herniation cases (60 dogs) (21 days compared to 3 days, respectively) and less frequent spinal pain during examination (20% compared to 75%, respectively). Spinal only meningoencephalomyelitis of unknown origin is an uncommon cause of myelopathy in French bulldogs that should be particularly considered in young adults with non-acute and non-painful presentation.

Modulation of 8-Oxoguanine DNA Glycosylase 1 (OGG1) Alleviated Anemia Severity and Excessive Cytokines Release during Plasmodium berghei Malaria in Mice.

The interplay of OGG1, 8-Oxoguanine, and oxidative stress triggers the exaggerated release of cytokines during malaria, which worsens the outcome of the disease. We aimed to investigate the involvement of OGG1 in malaria and assess the effect of modulating its activity on the cytokine environment and anemia during P. berghei malaria in mice. Plasmodium berghei ANKA infection in ICR mice was used as a malaria model. OGG1 concentration and oxidative stress levels in P. berghei-infected mice and their control counterparts were assessed during malaria using enzyme-linked immunosorbent assay. OGG1 activity in malaria mice was modulated using treatment with TH5487 and O8-OGG1 inhibitors. The effects of modulating OGG1 activity using OGG1 inhibitors on cytokine release and anemia during P. berghei malaria infection were assessed by cytometric bead array and measurement of total normal red blood cell count respectively. The plasma OGG1 level was significantly upregulated and positively correlated with parasitemia during P. berghei malaria in mice. Modulation of OGG1 ameliorated malaria severity by improving the total normal RBC count in TH5487 and O8-treated mice. Modulation of OGG1 with TH5487 caused significant reductions in serum levels of TNF-α, IFN-γ, IL-6, and IL-10. Similarly, OGG1 modulation activity using an O8-OGG1 inhibitor caused a significant reduction in serum levels of TNF-α, IL-2, IL-6, and IL-10. The findings indicate the involvement of OGG1 in the P. berghei malaria infection. OGG1 inhibition by TH5487 and O8-OGG1 inhibitors suppressed excessive cytokine release, and this may represent a novel therapeutic strategy for ameliorating the severity of malaria infection.

Effect of the added plasma rinseback on residual cell types in plateletpheresis leukoreduction systems.

Platelets collected by the Trima Accel apheresis device (Terumo BCT) are automatically leukoreduced through a leukoreduction system (LRS) where WBCs are trapped in a conical-shaped LRS chamber. The content has been used as a valuable source of mononuclear cells for research purposes. In frequent, long-term platelet apheresis donors, lymphopenia has been associated with the use of LRS chambers, and implementation of plasma rinseback at the end of the procedure has been shown to mitigate the depletion of lymphocytes. In this report, the cellular content of the LRS chamber and remaining disposable was characterized with and without plasma rinseback. Trima disposable sets were obtained from apheresis platelet collections in 100% plasma or 35% plasma/65% PAS with or without plasma rinseback at the end of the collections. Cellular content was drained from the LRS chamber and the disposable and was characterized using a hematology analyzer and flow cytometer to establish total cell counts and proportions of RBC, platelet, and WBC subpopulations. LRS chambers contained approximately 109 WBCs, with the majority being lymphocytes and monocytes. The addition of plasma rinseback significantly decreased the number of WBCs remaining in the disposable, thereby increasing the number of WBCs returned to the donor. However, rinseback did not impact the WBC content of the LRS chamber itself. Blood Centers using the Trima Accel instrument may reduce lymphopenia in regular platelet donors by implementing plasma rinseback, while ensuring the cellular content of the LRS chamber intended for research purposes remains unaffected.

Equine Asthma Diagnostics: Review of Influencing Factors and Difficulties in Diagnosing Subclinical Disease.

This literature review focuses on diagnostics of equine asthma (EA), possible influencing factors on diagnostic techniques and latest developments in diagnosing horses during EA remission or with subclinical disease. Routine EA diagnostics include a clinical examination of the respiratory system with percussion and auscultation including a rebreathing examination, and clinical pathology including white blood cells and arterial blood gas analysis. Subsequent diagnostics include bronchoscopy to evaluate the amount and viscosity of respiratory secretion, bronchoalveolar lavage, and the cytology of tracheal aspirates (TAs) and bronchoalveolar lavage fluid (BALF). The grading of EA severity is built on respiratory effort at rest, which is increased in severe equine asthma. The inflammatory subtype is based on BALF cytology, while TA cytology helps to rule out previous bacterial infections. Different factors have an impact on the airways regarding the structure of the epithelium, cytology, and inflammatory markers possibly influencing the diagnosis of EA. Short-term exercise increases the total cell count and inflammatory mediators identified in the BALF of human patients, asymptomatic horses, and other species. Other factors involve cold or chlorinated air, long-term training effects, and concurrent additional respiratory disease, in particular exercise-induced pulmonary hemorrhage. As BALF cytology may be unremarkable during EA remission and low-grade disease, exercise tests and other factors stressing the bronchial epithelium may help to diagnose these patients.

Orexin B alleviates sepsis-associated lung injury through the attenuation of pulmonary endothelial barrier dysfunction by regulating the rho-associated coiled-coil containing protein kinase 2/zonula occludens-1 (ROCK2/ZO-1) axis.

Dysfunction of the alveolar endothelial barrier plays a crucial role in the pathogenesis of septic acute lung injury (ALI). orexin B is a neuropeptide derived from orexin neurons in the lateral hypothalamus and has multiple biological functions. However, the physiological function of orexin B in sepsis is less reported. Here, we aimed to explore the protective effects of orexin B in sepsis-induced ALI and its underlying mechanisms. In this study, we established an ALI in vivo animal model in mice using cecal ligation and puncture (CLP) and an in vitro ALI model using mouse lung microvascular endothelial cells (MLMECs) induced with lipopolysaccharides (LPS). The animal experiments involved four groups: Sham, Sham+orexin B, CLP, CLP+orexin B. First, our results demonstrate that the levels of serum orexin B but not orexin A were reduced in septic mice. Correspondingly, the expression of orexin type 2 receptor (OX2R), but not orexin type 1 receptor (OX1R), was reduced in the lung tissue of septic mice. Administration of orexin B decreased the mortality in sepsis mice and improved M-CASS scores. Hematoxylin-eosin (H&E) staining assay demonstrated that administration of orexin B ameliorated histopathological lung injury. orexin B was also found to inhibit the inflammatory response in the lung tissues of septic mice by reducing the expression of tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), and recombinant chemokine C-X-C-motif ligand 15 (CXCL15). Additionally, the total cell count and neutrophils in bronchoalveolar lavage fluid (BALF) were reduced by orexin B. Notably, orexin B alleviated vascular endothelial permeability in mice lung tissue by increasing the expression of the tight junction protein zonula occludens-1 (ZO-1) and occludin. In vitro experiments demonstrated that orexin B prevented LPS-induced endothelial permeability in mouse lung microvascular endothelial cells (MLMECs) by upregulating the expression of ZO-1 and occludin. These effects are mediated by rho-associated coiled-coil containing protein kinase 2 (ROCK2). Based on these findings, we conclude that orexin B alleviates sepsis-induced ALI by ameliorating endothelial permeability of lung microvascular endothelial cells.

A Population-Based Study of the Mediating Role of WBC, NEUT and PLT in the Relationship Between Triglyceride-Glucose Index and Urinary Albumin Excretion.

To assess the potential association between the TyG index and the risk of abnormal UACR. Additionally, we aimed to determine the role and degree of influence of inflammatory biomarkers between the TyG index and abnormal UACR. A cross-sectional study recruited 1021 participants from a health management center between 2021 and 2022. Logistic or linear regression models, as well as mediation analysis, were employed to investigate the associations between the TyG index, inflammatory biomarkers (total and differential white blood cell counts, platelet, mean platelet volume(MPV), C-reactive protein(CRP)), and the risk of abnormal UACR. The study included 1021 participants, of whom 55.0% were men. The median age (interquartile range [IQR]) was 61.0 (53, 70) years. In multivariable-adjusted logistic regression models, both with and without the inclusion of smoking, alcohol drinking, BMI, Lipid-lowering drugs using, TC, SUA, ALT, and AST as potential covariates, the TyG index was associated with the risk of UACR, both with the odds ratios (ORs) per 1-standard deviation (SD) increase were 1.32 (95% CI, 1.08-1.62) and 1.27 (95% CI, 1.05-1.52), respectively. This study also demonstrated a significant indirect effect of the TyG index on the risk of abnormal UACR through total white blood cell counts, neutrophil counts and platelet (P values < 0.05); The proportions mediated was 11.2%, 3.5% and 29.6% for each respective variable. Insulin resistance and inflammation are associated with an increased risk of kidney insufficiency. And indicators of inflammation weakly mediate insulin resistance and risk of kidney insufficiency.

Lysophosphatidylcholine 14:0 Alleviates Lipopolysaccharide-Induced Acute Lung Injury via Protecting Alveolar Epithelial Barrier by Activation of Nrf2/HO-1 Pathway.

Acute lung injury (ALI) is characterized by diffuse alveolar injury and acute non-cardiac pulmonary edema, with high morbidity and mortality. Lysophosphatidylcholine 14:0 (LPC14:0) has anti-inflammatory and anti-oxidative effects in sepsis and bacteremia. We hypothesized that LPC14:0 could be a potential treatment for ALI. Therefore, the effects of LPC14:0 on lung epithelial cells and the underlying mechanism on ALI were investigated. Lipopolysaccharide (LPS) was instilled intratracheally in vivo while the Murine Lung Epithelial-12 was stimulated by tert-butyl hydroperoxide (t-BHP) in vitro to induce the ALI model. In vivo, lung injury was evaluated by histopathological changes and pulmonary edema was assessed by wet/dry ratio. Evans blue infiltration in lung tissue, total protein content, total cell counts and inflammatory factors in bronchoalveolar lavage fluid were evaluated for alveolar permeability. In vitro, cell viability and cell death rate were assessed by cell counting kit-8 and Calcein-AM/PI stain respectively. The expression of ZO-1, Occludin, Nrf2, and HO-1 was evaluated by Western blot. LPC14:0 attenuated the LPS-stimulated lung injury and oxidative stress in vivo, and alleviated the t-BHP-induced cell damage in vitro. Moreover, LPC14:0 significantly inhibited the degradation of the tight junction proteins and activated the Nrf2/HO-1 signaling pathway both in vivo and in vitro. Mechanistically, ML385, the Nrf2 inhibitor, inhibited the protective effects of LPC14:0 on barrier function in vitro. This study first demonstrated that LPC14:0 mitigated LPS-induced ALI and the destruction of tight junctions, at least in part through up-regulation of the Nrf2/HO-1 pathway.

Interaction between human dental microbiome and the host gingival model

Objective: Interactions between the dental biofilm and host are important to study infectious oral diseases such as dental caries and periodontitis. Previous studies have reported the reaction of a host gingival model co-cultured with bacteria or biofilms. However, these studies only report results of a gingival model, and a biofilm co-cultured with a gingival model has never been investigated although the microbiome of dental biofilms is associated with maintenance of health and onset of infectious oral diseases. Therefore, we aimed to investigate the reaction of a gingival model and experimental biofilm. An in situ dental biofilm model, which enables biofilm formation in the oral cavity, was used to prepare experimental biofilms. Methods: The in situ biofilm was co-cultured with a reconstructed gingival model (RHG). We performed 16S rRNA sequence analysis, metagenomic function prediction analysis, total bacterial cell counts using quantitative polymerase chain reaction, and confocal laser microscopic observation in addition to gene expression and histological investigation of RHG. Results: We found a change in Veillonella abundance in the in situ biofilms co-cultured with RHG. Metagenomic function prediction analysis revealed that the activity of certain pathways associated with amino acid production, lactic acid fermentation, and glycolysis system significantly increased in biofilms co-cultured with RHG. Conclusion: The microbiome and functions of dental biofilms can be affected by host gingival tissues. Our findings will contribute to the establishment of a host–biofilm interaction model to understand the pathophysiology of oral infectious diseases.

The inhibitory influence of carvacrol on behavioral modifications, brain oxidation, and general inflammation triggered by paraquat exposure through inhalation

The current study investigated how carvacrol (C) can prevent behavioral and brain oxidative changes, along with systemic inflammation caused by inhaled paraquat (PQ). Control rats exposed to saline solution, whereas six rat groups were subjected to PQ aerosols at a concentration of 54mg/m3 in 16 days. The PQ-exposed groups received saline (PQ group), C at dosages of 20 (C-L) and 80mg/kg/day (C-H), dexamethasone at a dosage of 0.03mg/kg/day, pioglitazone at dose of 5 and 10mg/kg/day (Pio-L and Pio-H), and a combination of C-L + Pio-L. Various parameters were assessed following the end of the treatment duration. There were marked elevation in total and differential white blood cell counts (WBCs), and malondialdehyde levels in the blood, hippocampus, and cerebral tissue but, thiol, superoxide dismutase (SOD), and catalase (CAT) exhibited a notable decrease (p < 0.05 to p < 0.001). The escape delay and traveled distance exhibited enhancement, however, on the probe day, the duration spent in the target quadrant and the time taken to enter the dark room at 3, 24, 48, and 72hours post an electrical shock, showed a reduction in the PQ group (P<0.05 to P<0.001). Inhaled PQ-induced changes were significantly improved in C, Pio, Dexa, and C-L + Pio-L treated groups (P<0.05 to P<0.001). The effects of C-L + Pio-L on most measured variables were higher than C-L and Pio-L (P<0.05 to P<0.001). C improved PQ-induced changes similar to dexamethasone and C-L showed additive effects when administered in combination with Pio.

Impact of stormwater on biofilm density and microbial community composition in water distribution networks.

Harvesting of stormwater and injecting it into aquifers for storage and recovery during high water demand periods is a promising technology for augmenting conventional water reserves. However, little has been known on how stormwater impacts the biofouling of water distribution infrastructure. This study evaluated the effect on harvested and limestone aquifer treated stormwater on biofilm formation in a pilot distribution pipe network compared to an identical drinking water pipe rig. Coupons made of cement, copper and polyvinyl chloride (PVC) pipe materials were installed to each pipe rig and exposed to stormwater or drinking water. The total cell counts determined by flow cytometry on the pilot rig coupons were in the order of 105 to 107 cells/cm2 for both source waters and showed some variation over the duration of the study. The culturable cell counts were somewhat higher for stormwater exposed coupons than for coupons in mains water rig. The total number of thermotolerant coliforms was notably higher on coupons exposed to stormwater than on those exposed to mains water. Considerable differences were observed in the bacterial and eukaryotic communities on coupons made of various materials and exposed to mains water and stormwater using pyrosequencing. Moreover, seasonal variations were observed in community composition and diversity. A number of bacterial and eukaryotic families and genera harbouring potential human pathogens were detected in both mains water and stormwater systems, with larger numbers of genera observed in the latter indicating a potentially increased risk of exposure to pathogens with stormwater. The stormwater system also harboured sulfur reducers, and a greater diversity of iron oxidisers. A number of bacterial genera that contribute to nitrogen cycling were observed in both mains water and stormwater systems. A number of bacteria grazing eukaryotes were detected, indicating that the biofilm communities are quite dynamic and the abundance of bacteria is able to support higher level eukaryotes.

A comparison of veterinary classification schemes, serum ascites albumin gradient,and Boyer's criteria in discriminating transudates from exudates in ascitic dogs.

To assess the parameters utilised by and the diagnostic performance of two traditional veterinary classification schemes (named A and B) based on ascites total protein and total nucleated cell count, the Boyer's criteria based on ascites lactic dehydrogenase activity, its serum ratio and the serum total protein, a simplified Boyer's criteria based on ascites lactic dehydrogenase activity and serum total protein only, and finally the serum-ascites albumin gradient in discriminate the pathophysiological origin of the ascites formation in dogs. Cross-sectional study including 291 client-owned dogs with ascites. Ascites aetiology was used to classify the pathophysiology of its formation. Parameters measured and calculated included ascites total protein, ascites total nucleated cell count, ascites lactic dehydrogenase activity and its serum ratio, serum total protein, and the serum-ascites albumin gradient. There were 33 transudates due to decreased colloid osmotic pressure, 69 transudates due to increased hydrostatic pressure gradient, and 189 exudates. Simplified Boyer's criteria misclassified 16 of 291 ascites (94.5% accuracy; 95% confidence interval: 91.2 to 96.8) and Boyer's criteria misclassified 21 of 291 ascites (92.8% accuracy; 95% confidence interval: 89.2 to 95.5). The traditional veterinary classification scheme B misclassified 71 of 291 ascites (75.6% accuracy; 95% confidence interval: 70.3 to 80.4) and scheme A 130 of 291 (55.3% accuracy; 95% confidence interval: 49.4 to 61.1). Finally, the serum-ascites albumin gradient misclassified 100 of 291 ascites (65.6% accuracy; 95% confidence interval: 59.9 to 71.1). The Boyers' criteria and a simplified Boyer's criteria were highly accurate in discriminating exudates from transudates, while the other classification schemes had significantly less diagnostic value in doing so.

Repression of JAK2-STAT1 and PD-L1 by CEP-33779 ameliorates the LPS-induced decline in phagocytic activity of alveolar macrophages and mitigates lung injury in mice.

The role of the JAK2-STAT1/PD-L1 pathway in the phagocytic activity of alveolar macrophages (AMs) during LPS-induced acute lung injury in mice remains poorly understood. This study aims to explore whether the JAK2-STAT1/PD-L1 pathway is upregulated on AMs in LPS-induced mice acute lung injury and to further explore the impact of the JAK2-specific inhibitor CEP-33779 on the LPS-induced impairment of AMs phagocytic activity and lung injury. ALI was induced in mice via intratracheal administration of LPS, followed by intragastric administration of JAK2 inhibitor CEP-33779 suspension. Immunohistochemistry was conducted to assess PD-L1 expression in lung tissue, as well as p-JAK2, p-STAT1, and PD-L1 expression on AMs in bronchoalveolar lavage fluid (BALF) using immunofluorescence. Levels of TNF-α and IL-6, as well as protein concentration in BALF, were measured using enzyme-linked immunosorbent assay and Bicinchoninic acid assays, respectively. Hematoxylin-eosin staining and lung injury score were employed to evaluate pathological changes in mouse lungs. Total cell count in BALF was determined using a cell counter. Furthermore, western blot and immunofluorescence was conducted to assess the effect of JAK2 and STAT1 inhibitor on JAK2-STAT1 pathway activation and PD-L1 expression, while confocal microscopy with latex beads rabbit IgG FITC complex was used to observe MH-S cells phagocytic ability. The study revealed that LPS stimulation triggered the activation of the JAK2-STAT1 pathway and an upregulation of PD-L1 on AMs in both LPS-induced acute lung injury mice and MH-S cell lines. Moreover, treatment with the JAK2 and STAT1 inhibitor effectively reduced the activation of JAK2-STAT1 signaling, downregulated PD-L1 expression on AMs in BALF from LPS-induced ALI mice and LPS-stimulated MH-S cells, and significantly improved the LPS-induced reduction in phagocytic activity in MH-S cells. Most notably, CEP-33779 treatment significantly mitigated the pulmonary inflammatory response and lung injury in mice with LPS-induced ALI. Collectively, these findings imply that the JAK2-STAT1 pathway plays a role in the upregulation of PD-L1, which in turn is associated with the diminished phagocytic activity in LPS-induced AMs as well as lung injury. Furthermore, our study highlights that CEP-33779 treatment can effectively improve the reduced phagocytic activity of AMs and relieve lung injury induced by LPS through suppression of the JAK2-STAT1/PD-L1 pathway.

Anti-fibrotic effects of lisinopril (ACE inhibitor) and fasudil (ROCK inhibitor) in combination for canine corneal fibrosis invitro.

Corneal fibrosis is a leading cause of blindness in mammalian species and may result in compromised performance in sports and daily functions. This study evaluated the safety and anti-fibrotic effects of the FDA-approved drugs, angiotensin-converting enzyme inhibitor (ACE-I) lisinopril and rho-kinase inhibitor (ROCK-I) fasudil, alone and in combination, on the canine cornea using an established invitro model. To test the safety and efficacy of lisinopril and fasudil, primary canine corneal fibroblasts (CCFs) generated from donor corneas of healthy dogs (n = 20) were used. A series of dose-dependent and time-dependent assays with lisinopril (1-50 μM) and fasudil (1-10 nM) were performed. qRT-PCR, immunofluorescence (IF) staining, cell viability assay, cell proliferation assay, LIVE/DEAD viability/cytotoxicity assay, TUNEL assay, and total cell count were performed. A 25-μM lisinopril and 3-nM fasudil dose were safe, nontoxic, and optimal for therapeutic evaluations invitro. Treatments of lisinopril or fasudil, alone or in-combination, to CCFs grown in the presence of TGF-β1 (5 ng/mL) showed inhibition of myofibroblast formation based on phase-contrast microscopy. The qRT-PCR and IF studies showed a significant decrease in expression of profibrotic markers, including α-smooth muscle actin (α-SMA; p < .0001), fibronectin (FN; p = .0002), tenascin C (TNC; p < .0001), Collagen I (Col-I; p < .0001), Collagen IIIA1 (Co-IIIA1; p < .0001), and Collagen IV (Co-lV; p < .0001). An ophthalmic formulation consisting of lisinopril and fasudil may offer a safe and effective method to treat canine corneal fibrosis. Additional studies evaluating safety and efficacy of this formulation invivo are warranted.