Number Of Polymorphonuclear Leukocytes Research Articles

Clinical and pathological features of cerebrospinal meningitis caused by Pantoea agglomerans : a case report

BackgroundPantoea agglomerans (P. agglomerans) is a gram-negative bacterium that is commonly isolated from plant surfaces, seeds, and the environment. As an opportunistic pathogen, it can cause blood, urinary and soft tissue infections in immunocompromised patients. In central nervous system, P. agglomerans infection has been report in children and immune-compromised patients, however, infection by such bacterium in nontraumatized immune competent adults has not been reported. Here, we report a case of P. agglomerans cerebrospinal meningitis accompanied by positive anti-myeloperoxidase (MPO) antibody in a 49-year-old female who has a history of black fungus planting.Case presentationThe patient manifested with repeated fever, headache, generalized muscle pain, and neurological defects. Cerebrospinal fluid (CSF) tests revealed a moderately elevated number of polymorphonuclear leukocytes (50–193 × 106/L), low glucose levels (0.54–2.44 mmo1/L), and extremely high protein content (2.42–25.42 g/L). Blood tests showed positive anti-myeloperoxidase antibodies lasting for 1.5 year before turning negative. Spine MRI showed thickening and enhancement of the whole spinal meninges. CSF metagenomic next-generation sequencing (mNGS) revealed 75,189 specific DNA reads of P. agglomerans. The patient underwent spinal laminectomy due to meningeal adhesions. Pathological results revealed fibrous tissue proliferation, inflammatory infiltration with focal necrosis and calcification in the dura mater. The patient was successfully treated with sufficient antibiotics at 1-year follow-up.ConclusionsPeople should be alert to CNS infections caused by P. agglomerans which presented with relatively mild clinical symptoms at onset, especially for those who contucts relevant agricultural and forestry work. The CSF characterization of P. agglomerans meningitis is elevated multiple nuclei white blood cells, significantly reduced glucose content, and markedly increased protein level which may be related to the secondary spinal membrane adhesions.

Disruption of the Functional Activity of Neutrophil Granulocytes as a Risk Factor for the Development of Lung Damage in Pregnant Women with COVID-19.

Currently, the assessment of immune status in patients with COVID-19 is limited to determining the count of polymorphonuclear leukocytes and the phagocytic function of neutrophils, which is insufficient to understand the regulatory role of innate immunity cells in the development of pneumonia. However, no such studies have been conducted in pregnant women with COVID-19. The aim of this study was to investigate the functional state of neutrophil granulocytes in order to identify predictors of pneumonia severity risk in pregnant women with COVID-19. A clinical characterization of pregnant women with COVID-19 in addition to minimal and average lung changes was provided. The composition and ratio of morphological forms of leukocyte cells were studied. Cytochemical studies of neutrophil granulocytes were carried out and calculations of the mean cytological index (MCI) for succinate dehydrogenase, myeloperoxidase, and cationic proteins were performed. The number of NETs in blood smears was counted. Independent predictors of pneumonia severity in pregnant women with COVID-19 were calculated using regression analysis. The quality of the model was assessed using ROC analysis. In pregnant women with COVID-19 and an average volume of lung changes, the number of polymorphonuclear leukocytes (p = 0.03) and band neutrophils (p = 0.002) in the blood was significantly higher than in pregnant women with minimal lung changes. The MCI indicators of succinate dehydrogenase, cationic proteins, and myeloperoxidase in pregnant women with COVID-19 were reduced in relation to the control group (p < 0.0001). In blood smears of pregnant women with COVID-19 and an average volume of lung changes, the number of NETs increased (p = 0.002). Regression analysis showed that succinate dehydrogenase and NETs are independent predictors of pneumonia severity in pregnant women with COVID-19. Our study confirms the prognostic significance of low levels of neutrophilic succinate dehydrogenase and high levels of NETs in the blood of pregnant women with COVID-19. The combination of these two biomarkers is a significant reflection of the severity of pneumonia development in pregnant women with COVID-19. However, further research is needed to identify the mechanisms underlying this association.

Antinociceptive and anti-inflammatory potential, and chemical characterization of the dichloromethane fraction of Solanum lycocarpum (Solanaceae) ripe fruits by LC-DAD-MS

Ethnopharmacological relevanceSolanum lycocarpum A. St. Hil. (Solanaceae) is a species from the Brazilian Cerrado, exhibiting several medicinal properties, being used by the population in the treatment of ulcers, bronchitis, asthma and hepatitis, which involve inflammatory processes. Aim of this studyThis study aimed to chemically characterize the dichloromethane fraction (DCM), as well as verify its antinociceptive, anti-inflammatory and antioxidant potential. Materials and methodsThe DCM fraction was obtained by partitioning the ethanol extract. The chemical constituents of the DCM fraction were characterized by LC-DAD-MS. The DPPH and FRAP assays were used to evaluate the antioxidant potential. The carrageenan-induced paw edema model was used to assess the anti-inflammatory effects, and the inflammatory infiltrate was evaluated by qualitative and quantitative histological analyses. The antinociceptive action of the DCM fraction was evaluated by acetic acid-induced abdominal writhing test, formalin-induced nociception and hot-plate test. ResultsSteroidal alkaloids solasonine, solasodine and solamargine, as well as the alkaloid peiminine/imperialine and caffeoylquinic acids, were annotated in DCM fraction by LC-DAD-MS. The DCM fraction showed antioxidative action in the in vitro DPPH and FRAP tests, as well as an anti-inflammatory effect for the three evaluated doses of 30, 100 and 300 mg/kg in the fourth and sixth hours after the administration of carrageenan. The histological analyses evidenced considerably reduction in leukocyte migration and the number of polymorphonuclear leukocytes. The study also demonstrated antinociceptive activity for the DCM fraction, which reduced abdominal writhing at three concentrations evaluated, as well as a decrease in paw licking in the formalin-induced nociception test both in the neurogenic phase and the inflammatory phase, with greater effectiveness compared to the anti-inflammatory indomethacin. The DCM fraction also increased the latency time of the animals in the hot plate test 60 min after treatment, although it did not seem to involve the opioidergic system. ConclusionThis work evidenced that the dichloromethane fraction of S. lycocarpum fruit possesses antinociceptive and anti-inflammatory potential, which supports its use in folk medicine for management inflammatory conditions.

Protective effects of mouthwash formulations of &lt;i&gt;Syzygium polyantha&lt;/i&gt; (L.) and &lt;i&gt;Piper betel&lt;/i&gt; (L.) on oral microbiota-induced gingivitis

Using a combination of natural ingredients as a mouthwash was expected to have a synergistic effect in preventing gingivitis, a common oral disease. The objective of this study was to elucidate the anti-inflammatory effect of different proportions of mouthwash infusions: F1 (75% Syzygium polyantha and 25% Piper betle) and F2 (25% Syzygium polyantha and 75% Piper betle) on oral microbiota causing gingivitis. Twenty-four Rattus norvegicus were divided into four groups, and bacteria were injected into the periodontal sulcus. The anti-inflammatory effect was assessed by calculating the reduced number of polymorphonuclear (PMN) leukocytes. A cytotoxicity test was carried out on the normal fibroblast cell line 3T3-L1. There were no significant differences in the decreased number of PMN leukocytes (p = 0.079 &gt; 0.05). Both F1 and F2 showed results of cell viability approaching 100% of living cells at concentrations of 0.29 ppm and 0.04 ppm, equivalent to 0.058% and 0.029%, respectively. This study concluded that both formulations of Syzygium polyantha and Piper betle have potential effects on gingivitis prevention. They had an effectiveness level almost similar to Chlorhexidine gluconate 2%. The toxicity value of formulation F1 is superior to that of formulation F2. Further studies concerning the toxicity of the mixtures and their effect on oral biofilm are needed.

Microvesicles and neutrophil aggression factors in the pathogenesis of chronic periodontitis

Background. It is known that the number of polymorphonuclear leukocytes increases with the development of periodontitis. However, their role in the pathogenesis of chronic periodontitis has not been fully defined.&#x0D; Aim. To study the pathogenetic role of polymorphonuclear leukocytes in the development of chronic periodontitis, realized by microvesicles and aggression factors (metalloproteinases, myeloperoxidase, calprotectin, lipocaine 2).&#x0D; Material and methods. 30 patients aged 30 to 50 years with a diagnosis of chronic generalized periodontitis of moderate severity (K05.3) and no severe somatic pathology were under observation. The comparison group included 20 people with no inflammatory diseases in the oral cavity, comparable with the main group in terms of gender, age, nationality, and the presence of bad habits. Using a standard set of indices, the state of periodontal tissues was assessed, and all examined patients underwent X-ray examination. In the oral fluid of the examined, the total number of microvesicles and their phenotype, the concentration of matrix metalloproteinases-2 and -9, myeloperoxidase, calprotectin, lipocaine 2 were determined. The parameters were determined using a flow cytometer. Descriptive statistics are represented by the median and interquartile range (25th and 75th percentiles). Two independent sample sets were compared using the MannWhitney test. Correlation analysis was carried out according to the Spearman method.&#x0D; Results. The total number of microvesicles in the oral fluid in patients with chronic periodontitis was 11 times higher than in healthy volunteers (p=0.00001). More than a third of them had neutrophil markers. The levels of neutrophil aggression factors were also higher in patients with periodontitis and correlated with the value of the periodontal index, the depth of the periodontal pocket, bleeding index, indicators of periodontal health. Thus, a strong positive relationship was found between the total number of microvesicles and the value of the periodontal index (r=0.675), the depth of the periodontal pocket (r=0.763), the bleeding index (r=0.704); the number of neutrophilic microvesicles and indicators of periodontal health r=0.816, r=0.837 and r=0.776, respectively. The content of metalloproteinase-2 correlated with the depth of the periodontal pocket (r=0.754), bleeding index (r=0.811), papillary-marginal-alveolar index (r=0.675).&#x0D; Conclusion. Neutrophils contribute to the pathogenesis of chronic periodontitis through the secretion of aggression factors: metalloproteinases, myeloperoxidase, calprotectin, lipocaine 2, and microvesicles.

Association Between the Level of Inflammation at Each Anatomical Sexual Activity from Gram Staining and Neisseria gonorrhoeae and Chlamydia trachomatis Infections

Objective: This study aimed to determine the association between the severity of inflammation at each anatomical sexual activity from gram staining with Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) infections. Materials &amp; Methods: This study was conducted using laboratory test data from patients at the Bangrak Sexually Transmitted Infections Center. The data obtained consisted of gram staining, which was divided by the number of polymorphonuclear leukocytes (PMNL), NG culture, and Nucleic Acid Amplification Test (NAAT) for NG and CT results. Results: For the diagnostic association between PMNL and NG infection, the results revealed that samples with urethral PMNL 3+ or 4+ carried a significant likelihood ratio (LR) for positive infection, LR 5.61 (P&lt;0.001) and LR 59.66 (P&lt;0.001), respectively. Cervical, rectal, and pharyngeal PMNL was not related to infection. For CT infection, urethral gram stains with PMNL levels were greater than or equal to 2+ and cervical specimens with PMNL 4+ were associated with CT infection. Rectal and pharyngeal PMNL showed no significant association with CT infection. Conclusion: Determination of PMNL levels from gram staining contributes to the diagnosis of patients with NG and CT in the urethra, particularly for patients with a high degree of inflammation.

EXPERIMENTAL STUDY OF THE POSSIBILITY OF DETERMINING THE OPTICAL DENSITY OF PLASMA OF VENOUS BLOOD TO MONITOR THE COURSE OF THE INTRA -ABDOMINAL INFLAMMATORY PROCESS

Early diagnosis of postoperative inflammatory intra-abdominal complications is an urgent issue due to the lack of sufficiently informative methods. Our previous studies revealed that measurement of venous blood plasma optical density parameters (ODVBP) at a wavelength of l = 310 nm is an informative criterion for the presence of inflammatory and destructive pathology of abdominal organs. Therefore, it would be logical to assume that such an indicator can be used to assess the activity of the intra-abdominal inflammatory process after surgery.&#x0D; The purpose of the study: to assess the possibility of determining the ODVBP at a wavelength of l = 310 nm to assess the activity of the intra-abdominal inflammatory process after surgery in the experiment.&#x0D; Materials and methods. 50 white non-linear rats with models of acute peritonitis, which was modeled by intra-abdominal puncture injection of a 20% autofecal mixture at a dose of 1 ml per 100 g of weight. 12 hours after the initiation of peritonitis, a laparotomy was performed, the peritoneal cavity was washed with a decamethoxin solution. After 6, 12, 24 and 48 hours, a relaparotomy was performed, the parietal peritoneum was taken for histological examination and blood from the jugular vein. The study of ODVBP was carried out on an Agilent Cary 100/300 Series UV-Vis spectrophotometer. Animals were removed from the experiment by decapitation. Inhalational sevoflurane anesthesia was used to anesthetize the animals. Statistical calculation of research results was carried out using Microsoft® Office Excel spreadsheets (build 11.5612.5703). Testing of the law of distribution of samples for normality was carried out using the Shapiro-Wilk test. The Wilcoxon test was used to test the hypothesis of equality of means.&#x0D; Results. 12 days after laparotomy, animals showed signs of purulent peritonitis. 6 hours after washing, the animals showed peritoneal edema, desquamation of the mesothelium, a small number of polymorphonuclear leukocytes (PLL) and lymphocytes. After 12 hours, the growth of peritoneal edema was observed, the desquamation of the mesothelium was maintained, and the infiltration of the peritoneal cavity of the peritoneal cavity increased. In the future, signs of subsidence of the inflammatory process in the peritoneum were observed.&#x0D; Against the background of the regression of inflammation 6 hours after washing the peritoneal cavity, the parameters of ODVBP decreased statistically significantly. After 12 hours, against the background of some increase in signs of inflammation, the indicators of ODVBP increased slightly. After 24 hours, against the background of signs of a moderate decrease in the activity of inflammation, the indicators of ODVBP decreased slightly. After 48 hours, against the background of a significant regression of signs of inflammation and the appearance of signs of regeneration processes, the indicators of ODVBP statistically significantly decreased, almost to the limits of normal values.&#x0D; The obtained data indicate that the determination of ODVBP is promising for use under clinical conditions for the purpose of control in the postoperative period. A decrease in parameters will indicate the absence of inflammatory complications, while an increase will indicate their possible development. However, this issue requires clinical research.&#x0D; Conclusions:&#x0D; &#x0D; According to experimental data, in rats with models of acute peritonitis, after washing the peritoneal cavity against the background of regression of inflammation of the peritoneum, ODVBP indicators at the wavelength l = 310 nm decrease statistically significantly.&#x0D; In case of activation of the inflammatory process, indicators of ODVBP increase.&#x0D; The obtained data indicate the prospects of clinical application of such an indicator for evaluating the activity of the inflammatory process and the prospects of relevant clinical studies.&#x0D;

Experimental study of intra-peritoneal instilation of recombinant human granulocyte colony-stimulating factor for the treatment of acute peritonitis

Aim. In the experiment, the possibility of intraperitoneal application of recombinant human granulocyte colony-stimulating factor (G-CSF) for the treatment of acute peritonitis (AP) was investigated.Materials and methods. 60 non-linear white rats. AP was modeled by intra-abdominal puncture of 20% autofecal mixture. After 12 hours, a laparotomy and sanitation of the peritoneal cavity were performed. In 30 animals (control), a solution of decamethoxine was used. In the experiment, after sanitation, a solution of G-CSF on NaCl was injected into the peritoneal cavity at a dose of 0.1 million units per 100 g of mass. After 6, 12, 24 and 48 hours, a relaparotomy was performed and the parietal peritoneum was taken for examination. The number of cells was counted on digital copies of histological preparations.The results. After 6 hours: in the control, the number of polymorphonuclear leukocytes (PLL) - 2.7±0.39, in the experiment - 3.9±0.38 (р&lt;0.05); in the control, the number of lymphocytes (LC) - 0.2±0.13, in the experiment - 0.7±0.33 (р&lt;0.05). After 12 hours: in the control, the number of PLL - 3.1±0.62, in the experiment - 4.6±0.45 (р&lt;0.05); in the control, the number of LC - 0-1 in the field of vision, in the experiment - 1.8±0.33; in the experiment, the number of fibroblasts (FB) was 0.9±0.27. After 24 hours: in the control, the number of PLL - 3.3±0.39, in the experiment - 1.3±0.33; in the control, the number of LC - 1.8±0.41, in the experiment - 2.3±0.33; in the control, the number of FB - 0.4±0.16, in the experiment - 1.6±0.31 (р&lt;0.01); in the control, the number of macrophages (MF) - 0.4±0.16, in the experiment - 0.7±0.21; in the experiment, the number of plasma cells (PC) was 1.1±0.28. After 48 hours: in the control, the number of PLL - 2.1±0.27, in the experiment - 1.4±0.31 (р&lt;0.05); in the control, the number of LC - 2.2±0.29, in the experiment - 3.3±0.37 (р&lt;0.05); in the control, the number of FB - 1.9±0.34, in the experiment - 2.9±0.23 (р&lt;0.05); in the control, the number of macrophages (MF) – 2.0±0.36, in the experiment – 3.4±0.22 (р&lt;0.01); in the control, the number of PCs was 0.9±0.28, in the experiment - 3.1±0.27 (р&lt;0.01). Conclusions. In animals with AP models, signs of a delay in the local response of immunocompetent cells for 12-24 hours, regeneration processes for 24-48 hours, and slowing down of the regression of the inflammatory process in the peritoneum are observed after the peritoneal cavity is cleaned with a decamethoxine solution. In animals that received intraperitoneal instillation of G-CSF after rehabilitation, signs of early activation of the local reaction of immune cells and regeneration processes are observed, along with the acceleration of the regression of inflammation. The results of the experiments indicate that intraperitoneal instillation of G-CSF can be used under clinical conditions for the treatment of patients with AP.

Formation and activity of NLRP3 inflammasome and histopathological changes in the lung of corpses with COVID-19.

COVID-19 is a contagious disease that attacks many organs but the lungs are the main organs affected. The inflammasome activation results in the exacerbation of inflammatory response in infectious disease. The aim of this study is to investigate the formation and activity of the NLRP3 inflammasome complex and the histopathological changes caused by the coronavirus in the lung of deceased persons with COVID-19. In total, 10 corpses; 5 corpses with no history of any infectious diseases and COVID-19 and 5 corpses with the cause of death of COVID-19 were included in this study. Lung tissue samples were harvested during autopsy under safe conditions. Fresh tissues in each group were used to measure the genes expression and proteins level of NLRP3, ASC, Caspase-1, IL-1β, IL-6 and TNF-α and a routine hematoxylin and eosin staining was performed for histological assessment. Data are represented as the means ± SD. Statistical significance difference was accepted at a p-value less than 5%. The NLRP3, ASC, Caspase-1, IL-1β, IL-6 and TNF-α genes expression and proteins level were elevated in the lung of the COVID-19 group in comparison with the control group. Histological findings presented the increasing number of polymorphonuclear leukocytes, macrophages and also pulmonary fibrosis in the lungs of corpses with the cause of death of COVID-19. High expression of NLRP3 inflammasome components and its relation with the pathophysiology of the coronavirus-infected lung suggested that targeting the NLRP3 inflammasome could be helpful in achieving a more effective treatment in patients with COVID-19.

Pulmonary toxicity and gene expression changes in response to whole-body inhalation exposure to multi-walled carbon nanotubes in rats

Purpose: To investigate the molecular mechanisms underlying the pulmonary toxicity induced by exposure to one form of multi-walled carbon nanotubes (MWCNT-7). Materials and methods: Rats were exposed, by whole-body inhalation, to air or an aerosol containing MWCNT-7 particles at target cumulative doses (concentration x time) ranging from 22.5 to 180 (mg/m3)h over a three-day (6 hours/day) period and toxicity and global gene expression profiles were determined in the lungs. Results: MWCNT-7 particles, associated with alveolar macrophages (AMs), were detected in rat lungs following the exposure. Mild to moderate lung pathological changes consisting of increased cellularity, thickening of the alveolar wall, alveolitis, fibrosis, and granuloma formation were detected. Bronchoalveolar lavage (BAL) toxicity parameters such as lactate dehydrogenase activity, number of AMs and polymorphonuclear leukocytes (PMNs), intracellular oxidant generation by phagocytes, and levels of cytokines were significantly (p < 0.05) increased in response to exposure to MWCNT-7. Global gene expression profiling identified several significantly differentially expressed genes (fold change >1.5 and FDR p value <0.05) in all the MWCNT-7 exposed rats. Bioinformatic analysis of the gene expression data identified significant enrichment of several diseases/biological function categories (for example, cancer, leukocyte migration, inflammatory response, mitosis, and movement of phagocytes) and canonical pathways (for example, kinetochore metaphase signaling pathway, granulocyte and agranulocyte adhesion and diapedesis, acute phase response, and LXR/RXR activation). The alterations in the lung toxicity parameters and gene expression changes exhibited a dose-response to the MWCNT exposure. Conclusions: Taken together, the data provided insights into the molecular mechanisms underlying the pulmonary toxicity induced by inhalation exposure of rats to MWCNT-7.

Gonoroični uretritis i rezistencija na antibiotike

Urethritis is a clinical syndrome characterized by the appearance of mucopurulent or purulent urethral secretions, with or without dysuria, due to an increased number of polymorphonuclear leukocytes in the anterior urethra. Recommended therapy for uncomplicated gonorrhea of the urethra, cervix and rectum when antimicrobial susceptibility of gonococci has not been tested, is a combination of ceftrixasone 1 g intramuscularly in one dose, with azithromycin 2 g in one oral dose. Antimicrobial therapy and prevention measures are the basis of the fight against bacterial urethritis. This fight is disturbed by antimicrobial resistance, which results in unsuccessful therapy and the possibility of complications of urethritis. Resistance of Neisseria gonorrhoeae to antibiotics has been known for decades, and there is a danger that in the future certain strains of N. gonorrhoeae will be resistant to all available antimicrobials if there are no new antibiotics that will not develop resistance quickly or in the case of development of effective vaccines.

Difference in the Number of Polymorphonuclear (PMN) Leukocytes of Gingival Crevicular Fluid (GCF) in Various Blood Types of Patients with Gingivitis

Difference in the Number of Polymorphonuclear (PMN) Leukocytes of Gingival Crevicular Fluid (GCF) in Various Blood Types of Patients with Gingivitis

Features of skin wound healing in rats with experimental chronic kidney disease

Chronic kidney disease negatively affects the morphofunctional state of all organs due to hemodynamic and metabolic disorders. Changes in the content of cytokines observed in kidney diseases, which regulate the processes of inflammation and tissue repair, can complicate the course of the wound process. This research aimed to study disorders in the process of skin wound repair due to changes in the dynamics of production of interleukins IL-1β, IL-6, IL-10, IL-4, growth factors bFGF and VEGF in animals with experimental chronic kidney disease. The levels of interleukins and growth factors were determined on the 7th, 14th and 28th days after surgical modeling of wounds in the blood of rats with experimental chronic kidney disease and animals of the control group. To assess the dynamics and quality of wound healing, a semi-quantitative histological analysis was performed. The study showed an increase in the content of pro-inflammatory interleukins in the group of sick rats: on the 7th day the level of IL-1β was 1.19 times higher, and IL-6 – 1.55 times, on the 14th day the level of IL-1β was 1.37 times in comparison with the control group. The maximum increase in the concentration of anti-inflammatory interleukins was noted on the 28th day: IL-4 was 2.10 times higher, IL-10 – 1.39 times higher than in the control group. The content of bFGF and VEGF in animals of the control group reached its maximum on the 7th day, and in animals with chronic kidney disease – on the 15th day after surgery. Semi-quantitative histological analysis showed a decrease in indicators in the group of sick animals: the number of fibroblasts and collagen deposition – on the 7th day, reepithelialization – on the 28th day. A persistent increase in the number of polymorphonuclear leukocytes was also noted at all periods of the experiment: by 1.38, 1.99, and 9.82 times – on the 7th, 14th, and 28th days, respectively. The study showed that the dynamics of the production of interleukins and growth factors were impaired in rats with chronic kidney disease. In the process of damage regeneration in sick animals, pro-inflammatory mechanisms prevailed with the involvement of a large number of immunocompetent cells, as a result, skin wounds took longer to heal.

Farelerde Oluşturulan Yeni Bir Alerjik Rinit Modeli

Objective: Allergic rhinitis (AR) is an inflammatory disease of the nasal mucosa mediated by IgE after exposure to an allergen. The most well known related comorbidity of AR is asthma. This study was planned due to the need for an animal model for studies on AR-asthma coexistence. In this study, the frequency of AR accompanying in the asthma model created in mice,and the usability of the related model in AR studies will be investigated. Methods: In our study, 6-8 week-old, 18-20 g BALB/c mice were used. Chicken egg ovalbumin (OVA Grade V, Sigma) was administered through intraperitoneal (IP) route at doses of 10 μg on days 0 and 14. Mice were exposed to aerosolized 2.5% ovalbumin solution in sterile saline for 30 minutes 3 days a week for 8 weeks, starting 7 days after the last IP administration (21st day). After exposure to OVA, mice were observed for typical signs of AR including sneezing, runny nose, and nasal itching. The final diagnosis of AR was made by histopathological examination of the rhinotracheal tissues of mice. Results: In our study, all mice exposed to ovalbumin received histopathologic diagnosis of AR. Increased number of capillaries lymphocytes, polymorphonuclear leukocytes and eosinophilsper square millimetre of rhinotracheal tissues were calculated in the murine model of AR compared to the the control group. Conclusion: This study introduced a new AR model, not cited in the literature, and induced with the longest-term ovalbumin exposure in the literature. It was concluded that this model, known as the asthma model, can also be used to induce an AR model and can be used in studies investigating coexistence of allergic rhinitis and asthma.

Surgical Suture Assembled with Tadalafil/Polycaprolactone Drug-Delivery for Vascular Stimulation Around Wound: Validated in a Preclinical Model

In this study, a novel Polycaprolactone suture assembled with Tadalafil was investigated to improve wound healing processes via vascular stimulation. Tadalafil/Polycaprolactone (TP) suture was developed by the electrospinning method. The designed suture was characterized by SEM, mechanical properties assessments, tensile strength measurements and the drug release study. For in vivo tests, rats were classified into two study groups. An incision was made on their back skin and they were sutured with TP suture and Polycaprolactone suture as control. Rats were sacrificed at 7 days following surgery for histopathological examinations with Hematoxylin and Eosin staining. Results of Tensile test demonstrated that the lowest tensile strength belonged to 3 and 4 % wt and the highest tensile strength belonged to 1 and 2 % wt of TP suture. The rate of Tadalafil release showed that the highest drug release was related to 3 and 4% wt which were about 125 to 210 μg during 15 days. The histopathology revealed that the number of blood vessels, collagen fibers, fibroblast, polymorphonuclear leukocytes, and epithelization was remarkable in Tadalafil/Polycaprolactone group during 7-day. A novel Tadalafil/Polycaprolactone suture improved the processes of wound healing by releasing the Tadalafil drug around the sutured wound and can be used in medical applications.

Cells and Cytokines in Milk of Subclinically Infected Bovine Mammary Glands after the Use of Immunomodulatory Composition GLP 810.

The aim of this study was to investigate the effect of intramammary infusions of natural composition GLP 810 with immunomodulating properties on the local nonspecific cellular and humoral immune response in cows with subclinical mastitis. The composition GLP 810 consists of lactic acid, lysozyme, glycopeptides, and 0.9% solution of NaCl. The following parameters were studied: (1) leukocyte differential distribution in milk, (2) expression of cytokines in milk leukocytes, (3) antibacterial activity, and (4) milk quality. Nineteen mammary glands in five lactating cows were infused with 10 mL of GLP 810, and nineteen other glands from five control cows were treated with 10 mL 0.9% NaCl. The results showed that after intramammary administration of the composition GLP 810 three times with 48 h intervals, the following effects on leukocyte populations in milk were observed: (1) an increase in the number of polymorphonuclear leukocytes and lymphocytes and (2) a decrease in the number of macrophages. A reduction in the number of pathogenic bacteria was also detected. The analyses of tumour necrosis factor-alpha, interleukin-10, and beta-defensin-2 revealed that the production of the aforementioned cytokines significantly increased, whereas no significant effects on interleukin-1 and caspase-6 expression in milk leukocytes were recorded. However, there were significant differences between mammary glands with high and low milk somatic cell count. The results suggest that the composition GLP 810 has an immunomodulatory effect on mammary glands and it could be used for improving the immune response in cows with subclinical mastitis during lactation.

Chronic Hepatitis E is associated with cholangitis.

Background and AimsSporadic hepatitis E is an emerging indigenous disease in Europe induced by genotype 3 of the virus. While the disease takes an acute self‐limited course in immunocompetent individuals, under immunocompromised conditions chronic hepatitis E might develop. The histology of chronic hepatitis E has not been described in detail systematically.MethodsLiver biopsies from 19 immunosuppressed patients with chronic hepatitis E were collected: 17 were organ transplant recipients, one had a CD4‐deficiency and one had received steroid therapy because of ulcerative colitis. Biopsies were processed with standard stains. Evaluation of histologic activity and fibrosis was performed according to Ishak. Additionally, immunohistochemistry with antibodies directed against open reading frame 2 and 3 of the virus was performed and liver biopsies were tested for hepatitis E virus RNA.ResultsBiochemical data showed an increase in alanine transaminase, aspartate transaminase, gamma‐glutamyl transferase and total bilirubin. Histopathology displayed typical features of chronic hepatitis with mild to moderate activity. The number of polymorphonuclear leucocytes was considerably increased and all patients had a florid cholangitis that presented as a destructive form in five of them. Hepatocytes and bile duct epithelia stained positive for hepatitis E virus by immunohistochemistry.ConclusionsChronic hepatitis E in immunocompromised individuals runs a similar course as hepatitis B and C and shows similar histopathology. However, the presence of destructive cholangitis in some cases accompanied by an increased number of polymorphonuclear leucocytes is markedly different. Immunohistochemically the virus is present in bile duct epithelia, seemingly the cause for cholangitis.

Cytokine and Adhesion Molecule Expression Induced by Different Strains of Staphylococcus aureus in Type 1 Diabetic Rats: Role of Insulin.

Introduction: Staphylococcus aureus may provoke peritonitis and death, especially in immunocompromized individuals such as diabetic patients. We evaluated the role of insulin in S. aureus-induced peritoneal infection in diabetic and non-diabetic rats.Materials/Methods: Alloxan-diabetic male Wistar rats and their respective controls received intraperitoneal injections of different strains of S. aureus or sterile phosphate-buffered saline. After 3 days of infection, the first set of diabetic and non-diabetic rats received 4 and 1 IU, respectively, of neutral protamine Hagedorn insulin and were analyzed 8 h later. The second set of diabetic and non-diabetic rats received 4 and 1 IU, respectively, of insulin 2 h before intraperitoneal infection and a half dose of insulin at 5 p.m. for the next 2 days and were analyzed 16 h later. The following measurements were performed: (a) number of cells in the peritoneal lavage fluid (PeLF), white blood cell count, and blood glucose; (b) serum insulin and corticosterone; (c) cytokine levels in the PeLF; (d) expression of adhesion molecules in the vascular endothelium; and (e) microbicidal activity.Results: Diabetic rats showed an increased number of polymorphonuclear leukocytes (PMNs) and increased concentrations of CINC-1, IL-4, and IFN-γ in the PeLF after infection with the ATCC 25923 or N315 αHL+ strain. The mesenteric expression of PECAM-1 was increased after infection with the N315 HLA+ strain. ICAM-1 expression was increased with ATCC infection. Treatment of diabetic rats with a single dose of insulin restored CINC-1 levels in the PeLF for both strains; however, PMN migration, IL-4, and IFN-γ were restored in rats infected with the ATCC strain, whereas the PeLF concentrations of CINC-2, IL-1β, and IL-4 were increased in N315-infected animals. Insulin restored PMN migration and CINC-2 levels in the PeLF in ATCC-infected rats. After multiple treatments with insulin, the levels of IL-1β, IL-6, and IFN-γ were increased in the PeLF of diabetic rats after infection with either strain, and CINC-2 levels were restored in N315-infected animals.Conclusion: These results suggest that insulin distinctively modulates cytokine production or release, PMN leukocyte migration, and adhesion molecule expression during the course of peritonitis induced by different strains of S. aureus.

Virulence factors and phylogenetic group profile of uterine Escherichia coli in early postpartum of high-producing dairy cows

Escherichia coli is the most common contaminant of the bovine uterus in the first few weeks postpartum, and one of the most important pathogens involved in uterine infections. This bacterium is characterised by diverse virulence factors (VF); however, the profile of E. coli VF in physiologic postpartum uterine contamination is not well established. Therefore, the present study aimed to investigate the prevalence of intrauterine bacteria, a set of virulence factors and phylogroups of E. coli isolates, leukogram and uterine cytology in 75 Holstein cows at 24 h (Sampling time 1) and 14 days (Sampling time 2) postpartum. Escherichia coli, α-haemolytic Streptococcus, Trueperella pyogenes, and Pasteurella multocida were the most common microorganisms isolated in pure culture, whereas E. coli and Clostridium perfringens Type A, E. coli and α-haemolytic Streptococcus, and E. coli and Proteus mirabilis were the most frequent microorganisms in mixed colonies at both studied sampling times. In the 59 E. coli isolates after 24 h (n = 35) and 14 days (n = 24) postpartum, the genes detected for VF were fimH, iucD/aer, kps, hlyA, usp, vt1 and vt2. Most E. coli strains detected in both moments of study belonged to Commensal phylogenetic groups A and B1, whereas Pathogenic groups D and B2 were identified at 24 h and 14 days postpartum respectively. All cows presented suitable immune response against the presence of bacteria in uterine lumen, observed by leukocytosis, neutrophilia, lymphocytosis and monocytosis at leukogram and a high number of polymorphonuclear leukocytes in uterine cytology, in both studied moments. Thus, a complex diversity of aerobic and anaerobic bacteria is involved in uterine contamination in the early postpartum of cows, besides the predominance of E. coli. Moreover, the genes fimH, iucD/aer, hlyA, kps, usp, vt1 and vt2 play a key role in the virulence of E. coli in this period.

Protective effects of ethyl gallate and pentagalloylglucose, the active components of Qingwen Baidu Decoction, against lipopolysaccharide-induced acute lung injury in rats.

BackgroundThe aim of this study was to investigate the bioactive constituents of Qingwen Baidu Decoction (QBD) in a rat model of acute lung injury (ALI) induced by lipopolysaccharide (LPS). Our previous studies showed that ethyl gallate (EG) and pentagalloylglucose (PGG) were the active components of QBD for the treatment of ALI.Materials and methodsWe isolated two compounds and identified the structures of them by nuclear magnetic resonance and mass spectrometer. Lung injury was induced by intratracheal instillation with LPS (5 mg/kg). Rats were randomly divided into six groups: Control group; LPS group; 5 mL/kg DEX + LPS group; 6.6 g/kg QBD extract + LPS group; 17.16 mg/kg PGG + LPS group; 7.26 mg/kg EG + LPS group. The effects of compounds on LPS-induced the number of total cells, neutrophils influx, protein leakage, W/D weight ratio and pulmonary histological changes were examined.ResultsThe results demonstrated that pretreatment with EG and PGG could notably inhibit lung edema and attenuate the pulmonary histological changes (P<0.05). The pretreatment also decreased the number of total cells and polymorphonuclear leukocytes in bronchoalveolar lavage fluid (BALF) (P<0.05).ConclusionEthyl gallate and pentagalloylglucose of QBD played a protective role in preventing LPS-induced ALI. The results supported further study of EG and PGG as potential candidates for preventing ALI.