Pouch Fluid Research Articles

Douglas Pouch Fluid Improves the Accuracy of Transvaginal Ultrasound in the Diagnosis of Uterosacral Ligaments Deep Infiltration Endometriosis: A Prospective Study.

To compare the accuracy of transvaginal ultrasound (TVUS) with laparoscopy in detecting and characterizing uterosacral ligament (USL) nodules of deep infiltrative endometriosis (DIE) between patients with and without pouch of Douglas (POD) fluid. This prospective study was conducted between June 2021 and July 2023. We included patients from the Second People's Hospital of Shenzhen with two TVUS tests: no POD fluid on the first TVUS test and POD fluid on the second TVUS test. POD fluid was obtained in two ways: naturally occurring fluid during the luteal phase of the menstrual cycle and SonoPODography. Laparoscopic results are the gold standard. To compare the diagnostic performance of TVUS in the diagnosis of DIE on USLs with and without POD fluid. We included 42 patients with a mean age of 36.4 ± 5.4 years. The maximum length diameter for DIE nodules on USLs with and without POD fluid was 13.3 ± 3.3 mm and 10.2 ± 2.5 mm (P < .001), respectively, while the depth of infiltration was 8.1 ± 2.4 mm and 6.1 ± 1.4 mm (P < .001), respectively. When compared to laparoscopic findings, TVUS findings with and without POD fluid resulted in a sensitivity, specificity, accuracy, positive predictive value, negative predictive value area under the curve, and Cohen kappa of 92.3%, 93.8%, 92.9%, 96.0%, 88.2%, 0.930, and 0.850 and 61.5%, 93.8%, 73.8%, 94.1%, 50.0%, 0.766, and 0.499, respectively. Laparoscopy showed USL involvement in up to 59.5% of patients. With or without fluid, the most common ultrasound features of DIE lesions were USL thickening, irregular morphology, clear boundaries, low echo, and no blood flow signal. For patients with clinical suspicion of DIE, TVUS with POD fluid has a higher accuracy for diagnosing USL DIE than TVUS without POD fluid.

Subacute Toxicity, Subacute Anti-inflammatory and Anti-arthritic Activities of Combination of Hydroethanolic Extract of Terminalia macroptera and Ximenia americana In-vivo

The aim of this study was to assess the anti-inflammatory effects of a combination of medicinal plants on two models of inflammation. Subacute toxicity was assessed by daily oral administration of 2000 mg/kg body weight (bw). Subacute inflammation and arthritis were induced using the carrageenan air pouch granuloma model and Complete Freund's Adjuvant (CFA) respectively. After 28 days of administration, the combination at 2000 mg/kg proved to be non-toxic and induced a significant reduction (p&lt;0.05) in transaminases and total cholesterol. The combinations C3 (150 mg/kg of T. macroptera + 250 mg/kg of X. americana), C2 ((250 mg/kg of T. macroptera + 150 mg/kg of X. americana) and C1 (250 mg/kg of T. macropteria + 250 mg/kg of X. americana) inhibited fresh granuloma formation by 40.37, 45.63 and 58.32% and dry granulomas by 47.77, 55.08 and 61.24% respectively. The combinations significantly (p&lt;0.001) reduced air pouch fluid volume and massive leukocytes infiltration compared with the control group. With regard to the anti-arthritic effect, the combination C1 showed significant inhibition (p&lt;0.05) of primary and secondary lesions compared with the control CFA. The increase in serum ALT, AST and uric acid concentrations observed in the CFA control group was significantly reduced (p&lt;0.001) by the combination C1. An antioxidant effect was observed with the administration of the combination C1 and prednisone, which resulted in a significant increase (P&lt;0.01) in GSH, SOD and catalase activity and a decrease in MDA concentration (P&lt;0.001) compared with the CFA control group.The results suggest that the combination C1 has anti-inflammatory and anti-arthritic effects and prevents oxidative stress in arthritic rats.

Sperm transport and male pregnancy in seahorses: An unusual model for reproductive science

The Syngnathidae (seahorses and pipefishes) are a group of teleost fishes in which, uniquely, developing embryos are hosted throughout pregnancy by males, using a specialized brood pouch situated on the abdomen or tail. Seahorses have evolved the most advanced form of brood pouch, whereby zygotes and embryos are intimately connected to the host's circulatory system and also bathed in pouch fluid. The pouch is closed to the external environment and has to perform functions such as gaseous exchange, removal of waste and maintenance of appropriate osmotic conditions, much like the mammalian placenta. Fertilization of the oocytes occurs within the brood pouch, but unlike the mammalian situation the sperm transport mechanism from the ejaculatory duct towards the pouch is unclear, and the sperm: egg ratio (about 5:1) is possibly the least of any vertebrate. In this review, there is highlighting of the difficulty of elucidating the sperm transport mechanism, based on studies of Hippocampus kuda. The similarities between seahorse pouch function and the mammalian placenta have led to suggestions that the pouch provides important nutritional support for the developing embryos, supplementing the nutritional functions of the yolk sac provided by the oocytes. In this review, there is a description of the recent evidence in support of this hypothesis, and also emphasis, as in mammals, that embryonic development depends on nutritional support from the placenta-like pouch at important stages of the gestational period ("critical windows").

Investigation of anti-inflammatory and anti-arthritic potentials of Terminalia catappa bark using in vitro assays and carrageenan-induced inflammation, complete Freund's adjuvant induced arthritis model in rats

Terminalia catappa Linn (T. catappa) belongs to the family Combretaceae, locally known as Bengal Almond. The plant is known to have many phytochemicals such as carbohydrates, phytosterols, tannins, polyphenols, flavonoids, and saponins. It is widely used in the treatment of various diseases such as dermatitis, leprosy, and scabies and internally for colic, headache and hepatitis and also, the plant possesses anticancer, antioxidant, antidiabetic, hepatoprotective and extracts have been shown to modify reproductive behavior in rats. The main objective of this study is to assess the pharmacological potentials of T. catappa bark in the therapeutic areas of inflammation and arthritis. T. catappa bark alcoholic extract (TCE) and water extract (TCW) were prepared and evaluated for its possible anti-inflammatory activity by in vitro protein denaturation assays and membrane stabilization assay. Further, the anti-inflammatory and anti-arthritic activity of bark extracts were evaluated by in vivo models such as carrageenan-induced paw edema, air-pouch model and Complete Freund's adjuvant (CFA) induced arthritis model in rats. Total phenolic content (TPC) was found to be 287 and 175 mg/g for TCE and TCW. Total flavonoid content (TFC) was found to be 10.2 and 61.7 mg/Kg for TCE and TCW. The concentration required for the 50 % inhibition of the protein denaturation (IC 50) was found to be lower for TCE (36.2 µg/mL) in egg albumin denaturation assay, whereas in the membrane stabilization assay method TCW exhibited a lower IC50 value in comparison with diclofenac. Both the extracts showed activity in the in vitro models of inflammation and were also found to be effective in reducing the paw edema in the carrageenan-induced inflammation model and inhibition of cellular infiltration and Myeloperoxidase activity in air pouch fluid, suggesting its anti-inflammatory activity. In CFA-induced arthritis model, the extracts exhibited anti-arthritic potential by improving the altered blood parameters, arthritic scores, radio-graphical, and histopathological changes. The results of the study suggested that the bark of T. catappa possessed anti-inflammatory and anti-arthritic activities.

MicroRNA-223 Deficiency Exacerbates Acute Inflammatory Response to Monosodium Urate Crystals by Targeting NLRP3.

ObjectiveMicroRNAs were identified as master-switch molecules limiting acute inflammatory response. This study investigated the potential role of microRNA (miR)-223 in the mechanism of gout.MethodsWild-type (WT) and miR-223 knock-out (KO) mice were used to evaluate the phenotypes of gout models. Inflammatory cytokines were measured in air pouch and peritoneal cavity lavage fluid. In addition to miR-223 level in gout patients, miR-223 and pro-inflammatory genes were examined in bone marrow-derived macrophages (BMDMs) from mice as well as peripheral blood mononuclear cells from healthy controls (HC) treated with monosodium urate (MSU) crystals in vitro.ResultsMiR-223 was up-regulated in the early phase in BMDMs from WT mice after MSU challenge and decreased rapidly, and this was not observed in miR-223 KO mice in vitro. In addition, miR-223 was required for macrophages homeostasis. In comparison with WT mice in vivo, miR-223 deficiency exacerbated swelling index of MSU-induced inflammation in foot pad and ankle joint models. MiR-223 deficiency also markedly aggravated inflammatory cells infiltration and cytokines release including interleukin (IL)-1β, IL-6 and monocyte chemotactic protein-1 (MCP-1) in the air pouch and peritonitis models. In the in vitro experiments, miR-223 deficiency promoted the inflammatory response by targeting NLR family pyrin domain containing protein 3 (NLRP3). Besides, miR-223 level was down-regulated in gout patients and in HC exposed to MSU in vitro.ConclusionMiR-223 was down-regulated in gout patients and miR-223 deficiency exacerbated inflammatory response in diverse murine models, suggesting that up-regulation of miR-223 could be a potential therapeutic strategy for alleviating gouty inflammation.

Biliverdin and bilirubin sulfonate inhibit monosodium urate induced sterile inflammation in the rat

BackgroundBiliverdin, a by-product of haem catabolism, possesses potent endogenous antioxidant and anti-inflammatory properties. Bilirubin-C10-sulfonate (BRS), an active metabolite formed after enteral administration of BV in the rat, also possess antioxidant properties. Therefore, we investigated the anti-inflammatory and antioxidant activity of BV and BRS in an in vivo model of monosodium urate induced sterile inflammation. MethodsSubcutaneous air pouches were created on the dorsal flanks of Wistar rats (10–12 weeks of age). Prior to stimulation of the 6-day old pouch with monosodium urate (25 mg), groups were pre-treated with intraperitoneal BRS (27 mg/kg) and BV (27 mg/kg). Total and differential leukocyte counts were determined in pouch fluid aspirate at 1, 6, 12, 24 and 48 h after monosodium urate stimulation. Biliverdin (BV), BRS and unconjugated bilirubin (UCB) concentrations in the serum and pouch fluid were quantified using liquid chromatography-mass spectrometry. Pouch fluid cytokine concentrations (IL-1β, IL-1α, TNF-α, IL-17A, IL-12, GM-CSF, IL-33, IFN-γ, IL-18, IL-10, MCP-1, CXCL-1 and IL-6) were assessed after 6 h. In addition, 24 h protein carbonyl and chloramine concentrations were assessed in pouch fluid using ELISA and spectrophotometry, respectively. ResultsBRS and BV significantly (p < 0.05) inhibited leukocyte (total, neutrophil and macrophage) infiltration into the pouch fluid from 6 to 48 h. For example, after 6 h neutrophil counts decreased following BRS (0.32 ± 0.11 × 106 cells mL−1) and BV (0.17 ± 0.03 × 106 cells mL−1) compared to MSU only (3.51 ± 1.07 × 106 cells mL−1). Both BV and BRS significantly (p < 0.05) reduced pouch GM-CSF (BV: 5.8 ± 1.2 pg mL−1, BRS: 6.9 ± 1.5 pg mL−1 vs MSU only: 13.0 ± 1.9 pg mL−1) and MCP-1 concentrations at 6 h (BV: 1804 ± 269 pg mL−1, BRS: 7927 ± 2668 pg mL−1 vs MSU only: 17,290 ± 4503 pg ml−1), whilst BV additionally inhibited IL-6 (4354 ± 977 pg mL−1 vs MSU only: 25,070 ± 5178 pg mL−1) and IL-18 (17.6 ± 2.0 pg mL−1 vs MSU only: 81.5 ± 19.9 pg mL−1) concentrations at 6 h (p < 0.05). Despite these differences, no change in pouch chloramine or protein carbonyl concentrations occurred at 24 h (p > 0.05). Serum BV concentrations rapidly diminished over 6 h, however, BRS was readily detected in the serum over 48 h, and in pouch fluid over 12 h. ConclusionsThis study is the first to elucidate anti-inflammatory activity of BRS and the efficacy of BV administration in a model of gouty inflammation. Reduced leukocyte infiltration and cytokine production in response to sterile inflammation further support the importance of these molecules in physiology and their therapeutic potential in sterile inflammation.

Arhalofenate acid inhibits monosodium urate crystal-induced inflammatory responses through activation of AMP-activated protein kinase (AMPK) signaling

BackgroundArhalofenate acid, the active acid form of arhalofenate, is a non-agonist peroxisome proliferator-activated receptor γ (PPARγ) ligand, with uricosuric activity via URAT1 inhibition. Phase II studies revealed decreased acute arthritis flares in arhalofenate-treated gout compared with allopurinol alone. Hence, we investigated the anti-inflammatory effects and mechanisms of arhalofenate and its active acid form for responses to monosodium urate (MSU) crystals.MethodsWe assessed in-vivo responses to MSU crystals in murine subcutaneous air pouches and in-vitro responses in murine bone marrow-derived macrophages (BMDMs) by enzyme-linked immunosorbent assay (ELISA), SDS-PAGE/Western blot, immunostaining, and transmission electron microscopy analyses.ResultsOral administration of arhalofenate (250 mg/kg) blunted total leukocyte ingress, neutrophil influx, and air pouch fluid interleukin (IL)-1β, IL-6, and CXCL1 in response to MSU crystal injection (p < 0.05 for each). Arhalofenate acid (100 μM) attenuated MSU crystal-induced IL-1β production in BMDMs via inhibition of NLRP3 inflammasome activation. In addition, arhalofenate acid dose-dependently increased activation (as assessed by phosphorylation) of AMP-activated protein kinase (AMPK). Studying AMPKα1 knockout mice, we elucidated that AMPK mediated the anti-inflammatory effects of arhalofenate acid. Moreover, arhalofenate acid attenuated the capacity of MSU crystals to suppress AMPK activity, regulated expression of multiple downstream AMPK targets that modulate mitochondrial function and oxidative stress, preserved intact mitochondrial cristae and volume density, and promoted anti-inflammatory autophagy flux in BMDMs.ConclusionsArhalofenate acid is anti-inflammatory and acts via AMPK activation and its downstream signaling in macrophages. These effects likely contribute to a reduction of gout flares.

ACUTE AND SUBACUTE ANTI-INFLAMMATORY ACTIVITIES OF DICHLOROMETHANE EXTRACT OF CASSIA ALATA (LINN.) LEAVES IN WISTAR RATS

Background: In Burkina Faso, leaves of Cassia alata are used in the treatment of several diseases, including inflammation. This study evaluated the anti-inflammatory effects of the dichloromethane extract of Cassia alata leaves (CF-AECal) on different models of inflammation in wistar rats in order to enhance its use in traditional medicine. &#x0D; Materials and Methods: Acute inflammation was induced among rats with 0.1 ml of carrageenan, serotonin, histamine and dextrane 1%. The effects of the CF-AECal 50 mg/kg and 100 mg/kg were compared to the effects of diclofenal 5 mg/kg, prednisone 5 mg/kg, promethazine 1 mg/kg, cyproheptadin 2 mg/kg. Cotton pellet and air pouch - induced granuloma permitted to study CF-AECal effects on the subacute inflammation. The ways of administrations were intra peritoneal for the substances of inductions and oral for the molecules of references and CF-AECal. &#x0D; Result: Carrageenan-induced paw edema has been inhibited to 52.39% by CF-AECal 50 mg/kg and 50.17% by CF-AECal 100 mg/kg. Wet granulomas formation has been inhibited by CF-AECal 50 and 100 mg/kg to 20.94 and 57.82%. The dry granulomas were inhibited at 24.32 and 59.04% by CF-AECal 50 and 100 mg/kg. Air pouch fluid was significantly reduced by CF-AECal in comparison to the control group. There was a massive recruitment of leukocytes in the pocket granuloma of the control group. CF-AECal caused strong inhibition of this infiltration of leukocytes (p&lt; 0.001). &#x0D; Conclusion: Dichloromethane extract of Cassia alata showed anti-inflammatory effects.

Embryo oxygenation in pipefish brood pouches: novel insights

The pipefish brood pouch presents a unique mode of parental care that enables males to protect, osmoregulate, nourish and oxygenate the developing young. Using a very fine O2 probe, we assessed the extent to which males of the broad-nosed pipefish (Syngnathus typhle) oxygenate the developing embryos and are able to maintain pouch fluid O2 levels when brooding in normoxia (100% O2 saturation) and hypoxia (40% O2 saturation) for 24 days. In both treatments, pouch fluid O2 saturation levels were lower compared with the surrounding water and decreased throughout the brooding period, reflecting greater offspring demand for O2 during development and/or decreasing paternal ability to provide O2 to the embryos. Male condition (hepatosomatic index) was negatively affected by hypoxia. Larger males had higher pouch fluid O2 saturation levels compared with smaller males, and levels were higher in the bottom section of the pouch compared with other sections. Embryo size was positively correlated with O2 availability, irrespective of their position in the pouch. Two important conclusions can be drawn from our findings. First, our results highlight a potential limitation to brooding within the pouch and dismiss the notion of closed brood pouches as well-oxygenated structures promoting the evolution of larger eggs in syngnathids. Second, we provide direct evidence that paternal care improves with male size in this species. This finding offers an explanation for the documented strong female preference for larger partners because, in terms of oxygenation, the brood pouch can restrict embryo growth.

93. Macrophages Are a Barrier to AAV5 Mediated Gene Expression in the Air Pouch Synovial Inflammation (APSI) Model

Our group is investigating the application of AAV gene therapy for inflammatory disease, especially rheumatoid arthritis (RA). The air pouch synovial inflammation (APSI) model is commonly used for studying local inflammation in rodents and is often used to test anti-rheumatic compounds. It involves the subcutaneous injection of air in the back of the animal, promoting the formation of an air pouch lining. After ~6 days, this lining bears a strong resemblance to the synovial membrane of a joint, being composed primarily of fibroblast-like and macrophage-like cells. Inflammation can be induced by injecting a stimulus (e.g. LPS, TNF, synovial fluid) into the air pouch cavity. While not a true RA model, the APSI model has a number of advantages over standard mouse arthritis models, including a short time frame (2 weeks versus 2 months), low discomfort for the animals, and allows for analysis of inflammatory markers in the air pouch fluid and membrane. Additionally, many of the treatments that have proved effective in human RA (e.g. methotrexate, TNF-blockers, steroids) are also effective in the APSI model.In order to use the APSI model for testing new gene therapies for RA, we investigated the kinetics of AAV5 mediated gene expression. An AAV5-CMV-Luciferase vector was used to determine optimal vector doses by non-invasive in vivo whole animal bioluminescent imaging. To our surprise, when vector was injected at day 6 after formation of the air pouch, we were unable to detect any luciferase expression up to day 30, over a wide range of vector doses (1×10E9 to 1×10E12 vg). Subsequent studies confirmed that similarly to synovial fibroblasts, air pouch fibroblasts were able to be transduced by AAV5 in vitro.As immunohistochemistry (IHC) staining (F4/80) revealed a high density of macrophages in the air pouch lining at day 6, we hypothesized that the presence of these cells might be inhibiting AAV5 mediated transduction in the APSI model. To test this hypothesis we either suppressed macrophage activity using triamcinolone (steroid) in combination with AAV5-CMV-Luciferase injection at day 6, or avoided macrophages by administering vector before the influx of macrophages into the air pouch lining (day 0). We observed that either suppressing or avoiding macrophages allowed for expression of luciferase following AAV5-CMV-Luciferase administration.These data suggest that macrophages are a previously underappreciated barrier to gene transfer, and that strategies, such as immune suppression or macrophage depletion, should be explored further when administering vector to macrophage rich tissue.

Evidence of impairment of normal inflammatory reaction by a high-fat diet

The murine dorsal air pouch model is a valuable tool for studying acute peripheral inflammatory reactions. We used this model to study the effect of diet on the onset of acute inflammation. Mice were fed a normal or a high-fat diet (HFD) for 5 weeks. Air pouches were raised and injected with non-stimulating (saline) or stimulating solution (saline containing lipopolysaccharides). After 4 h, leukocytes in the pouch fluid were enumerated, sorted and their viability measured. Cytokine/chemokine levels in the cell-free fluid were measured using a cytometric bead assay. Gene expression level was measured in leukocytes and in lining tissues using comparative real-time PCR. Leukocyte migration and cytokine/chemokine secretion were decreased substantially in mice fed the HFD. In contrast, leptin levels were elevated. Gene expression profiles in leukocytes recovered from the pouch and in the pouch-lining tissue (believed to have an important role in the initiation of granulocyte recruitment) were depressed. Genes encoding CC and CXC family chemokines were among the most negatively affected. These results suggest that a HFD can alter peripheral tissue activation as well as leukocyte recruitment and response, thereby affecting the development of an effective local immune response, which could have deleterious consequences.

A mouse air pouch model for evaluating the immune response to Taenia crassiceps infection

The experimental system of Taenia crassiceps cysticerci infection in BALB/c mice is considered to be the most representative model of cysticercosis. In our work, mice were sacrificed 7 and 30days after infection, and pouch fluid was collected to determine the number of accumulated cells and the concentrations of IFNγ, IL-2, IL-4, IL-6, IL-10 and nitric oxide. The injection of 50 nonbudding cysticerci into normal mouse dorsal air pouches induced a high level of IFNγ and nitric oxide production relative to the parasite load. The air pouch provides a convenient cavity that allows studying the cellular immunological aspects of the T. crassiceps parasite. The nonbudding cysticerci recovered from the air pouches contained cells that can reconstitute complete cysts in the peritoneal cavity of mice. In conclusion, these results demonstrate that the air pouch model is an alternative tool for the evaluation of the immune characteristics of T. crassiceps infection.

Natural Honey: A New and Potent Anti-Angiogenic Agent in the Air-Pouch Model of Inflammation

Despite reports indicating anti-inflammatory effects of honey, the anti-angiogenic effect of honey and its impact on inflammatory mediators in the air pouch model of inflammation have not yet been studied. The aims of present study were to investigate the effects of honey on angiogenesis, inflammatory cytokine vascular endothelial growth factor (VEGF) level as an important marker of angiogenesis and prostaglandin E2 (PGE2) in the rat air pouch model of inflammation. Male Wistar rats were anesthetized, and then 20 ml and 10 ml of sterile air were injected subcutaneously in the back on days 0 and 3, respectively. On day 6, inflammation was induced by injection of 1 ml of carrageenan 1% into pouches. After 72 h, the rats were sacrificed; pouch fluid was collected in order to determine PGE2 concentration and VEGF level. The Pouches were dissected out and weighed. Angiogenesis of granulomatous tissue was assayed using a hemoglobin kit. Honey was able to reduce granulation tissue weight and angiogenesis as well as showing potent inhibitory activities against PGE2 and VEGF in air pouch model of inflammation. The decrease in angiogenesis correlates with the inhibition of PGE2 and VEGF. Honey is potentially useful in the treatment of granulomatous inflammatory conditions. It seems that the anti-angiogenic activities of honey are mediated through modulation of PGE2 and VEGF production.

Colonization of the lower urogenital tract with Ureaplasma parvum can cause asymptomatic infection of the upper reproductive system in women: a preliminary study

PurposeGenital ureaplasmas are considered opportunistic pathogens of human genitourinary tract involved in adverse pregnancy sequelae and infertility. While association of Ureaplasma urealyticum with urogenital tract infections is well established, the role of Ureaplasma parvum in these infections is still insufficient. In the study, we compared how often cervicovaginal colonization with U. parvum is associated with the presence of these microorganisms in the upper genitourinary tract of fertile and infertile women.MethodsWe used PCR assay to determine the prevalence of U. parvum and U. urealyticum in pairs of specimens, i.e., vaginal swabs and Douglas’ pouch fluid samples from consecutive 40 women with no symptoms of genital tract infection.ResultsIn total, 19 (47.5 %) of the 40 samples were positive for ureaplasmas. U. parvum was simultaneously detected in pairs of samples in five (55.5 %) of the nine (47.4 %) women positive in PCR assay. As many as 5 (18.5 %) of the 27 infertile women and 1 (7.7 %) of the 13 fertile women showed infection of the upper genital tract with U. parvum.ConclusionThe results of the study demonstrated that colonization of the lower genital tract with U. parvum can produce asymptomatic infection of the upper reproductive system in women. These findings also imply that U. parvum may be present in the upper genital tract at the time of conception and might be involved in adverse pregnancy outcomes.

Preparation, macrophages targeting delivery and anti-inflammatory study of pentapeptide grafted nanostructured lipid carriers

The targeting ability of pentapeptide (Thr-Lys-Pro-Pro-Arg) grafted nanostructured lipid carriers (Pen-NLCs) to macrophages was investigated in both in vitro and in vivo studies. The results showed the improvement of the anti-inflammatory effect by using this drug delivery system. Firstly, a pentapeptide-polyethylene glycol2000-stearate was synthesized and formulated into Pen-NLCs. Non-grafted nanostructured lipid carriers (Bare-NLCs) and Pen-NLCs were 190.0±1.0 and 203.0±8.5 nm in size, -8.1±2.1 and 2.3±1.2 mV in zeta potential respectively. Meanwhile, they had comparable entrapment efficiency and drug loading efficiency. In vitro and in vivo cellular uptake studies showed increased internalization of Pen-NLCs by macrophages when compared to pure drugs and Bare-NLCs. Animal studies in a carrageenan-treated air pouch model were used to further investigate the anti-inflammatory effects of Pen-NLCs. Through intravenous administration, a single dose of DXM loaded Pen-NLCs showed the strongest inhibition of inflammatory indexes of air pouch fluid weight, leukocyte infiltration, granulation tissue weight and nitric oxide concentration in comparison with free drugs and DXM loaded Bare-NLCs. In conclusion, this study demonstrated the potential of Pen-NLCs as promising drug carriers for anti-inflammatory treatments.

Perturbation of Staphylococcus aureus Gene Expression by the Enoyl-Acyl Carrier Protein Reductase Inhibitor AFN-1252

This study examines the alteration in Staphylococcus aureus gene expression following treatment with the type 2 fatty acid synthesis inhibitor AFN-1252. An Affymetrix array study showed that AFN-1252 rapidly increased the expression of fatty acid synthetic genes and repressed the expression of virulence genes controlled by the SaeRS 2-component regulator in exponentially growing cells. AFN-1252 did not alter virulence mRNA levels in a saeR deletion strain or in strain Newman expressing a constitutively active SaeS kinase. AFN-1252 caused a more pronounced increase in fabH mRNA levels in cells entering stationary phase, whereas the depression of virulence factor transcription was attenuated. The effect of AFN-1252 on gene expression in vivo was determined using a mouse subcutaneous granuloma infection model. AFN-1252 was therapeutically effective, and the exposure (area under the concentration-time curve from 0 to 48 h [AUC(0-48)]) of AFN-1252 in the pouch fluid was comparable to the plasma levels in orally dosed animals. The inhibition of fatty acid biosynthesis by AFN-1252 in the infected pouches was signified by the substantial and sustained increase in fabH mRNA levels in pouch-associated bacteria, whereas depression of virulence factor mRNA levels in the AFN-1252-treated pouch bacteria was not as evident as it was in exponentially growing cells in vitro. The trends in fabH and virulence factor gene expression in the animal were similar to those in slower-growing bacteria in vitro. These data indicate that the effects of AFN-1252 on virulence factor gene expression depend on the physiological state of the bacteria.

Characterization of the inflammatory response to four commercial bone graft substitutes using a murine biocompatibility model

Bone grafting is utilized in nearly all orthopedic subspecialties and in most anatomic regions. Bone graft substitutes have the potential to offer similar efficacy as autogenous grafts without the morbidity of harvest. Several studies have noted the efficacy of new-generation bone substitute products, but few studies have evaluated their safety. This study characterizes and quantifies the inflammatory reaction to four different commercially available bone graft substitutes, which were examined using the in vivo murine air pouch biocompatibility model. One coralline hydroxyapatite product was chosen as an example of a purely osteoconductive material. Three demineralized bone matrix products were chosen to represent products that are both osteoconductive and osteoinductive. Samples were implanted in a murine air pouch and harvested after 14 days in situ. Pouch fluid was extracted, mRNA isolated, and reverse transcription polymerase chain reactions carried out to detect interleukin-1 gene expression as a marker for inflammation. In addition, multiple histological characteristics were examined to quantify cellular responses to the implanted materials. All bone graft substitutes induced a significant inflammatory response compared with negative controls. Histology and polymerase chain reaction data indicated that the level of inflammatory reaction was elevated in materials with a higher demineralized bone matrix to carrier proportion. The hydroxyapatite product generated a low inflammatory reaction. In conclusion, this study used an in vivo model of biocompatibility to demonstrate that a significant inflammatory reaction occurs when using implanted bone graft substitutes. When choosing a bone grafting method, surgeons should consider both the efficacy and safety of methods and materials used. Further studies are necessary to determine the ideal bone graft material to maximize efficacy while minimizing morbidity.

Anti-inflammatory activity of Ajmodadi Churna extract against acute inflammation in rats

Background:Ayurvedic polyherbal formulations are widely prescribed for a wide range of inflammatory conditions, yet, despite widespread use, there has been no systematic documentation of their safety and efficacy.Objective:The present study was undertaken to evaluate the anti-inflammatory activity of aqueous extracts of Ajmodadi churna (AJM) in rats.Materials and Methods:Carrageenan-induced hind paw edema and air pouch inflammation models were used for the study.Results:The extracts showed significant antiinflammatory activity, reducing paw edema volume by 0.417 ± 0.097 and 0.379 ± 0.049, respectively. In the carrageenan-induced air pouch model, AJM reduced total leukocyte count by 73.09 ± 7.13 and 62.17 ± 10.53, granulocyte count by 69.48 ± 5.44 and 63.33 ± 4.13, and myeloperoxidase activity by 14.84 ± 0.91 and 18.44 ± 3.18, respectively, compared to controls.Discussion and Conclusion:AJM significantly reduced paw edema, during the second phase of edema development. In the carrageenan-induced air pouch model, AJM inhibited cellular infiltration into the air pouch fluid. We conclude that AJM is an effective candidate for prevention or treatment of acute inflammation

Albumin coupled lipid nanoemulsions of diclofenac for targeted delivery to inflammation

Diclofenac lipid nanoemulsions (DLNEs) were prepared with different compositions. Based on size, PDI, zeta potential, and in vitro drug release, the optimized DLNEs (DLNE-4 and DLNE-7) were developed and evaluated for drug content, entrapment efficiencies, and stability in comparison to the control formulation (DLNE-1). The albumin was coupled to DLNE-7 globules (DLNE-8) by water soluble carbodiimide (EDC) method, purified, and quantified by modified Bradford method. The pharmacokinetic study was conducted in inflammation (granuloma air pouch model) induced rats. The maximum peak concentration of DLNE-8 was almost fourfold to fivefold in comparison to drug solution in granuloma air pouch fluid (GAPF). The therapeutic availability (TA) of DLNE-8 was 2.89, 2.34, and 1.66 times that of drug solution, DLNE-4 and DLNE-7, respectively. The GAPF/serum ratio of diclofenac from DLNE-8 was above one at all time points indicating the targeting potential of albumin ligated LNEs to inflammatory sites. From the Clinical EditorThis study demonstrates targeted delivery of diclofenac to an inflammatory environment using the granuloma air pouch model and diclofenac nanoemulsions with different compositions.

Use of 3-hydroxy fatty acid concentrations in a murine air pouch infection model as a surrogate marker for LPS activity: A feasibility study using environmental Burkholderia cenocepacia isolates

Using a murine hypodermic air pouch infection model designed to mimic the release of bacterial products at physiological levels, 3-hydroxy fatty acid (3-OH FA) and endotoxin unit levels from Burkholderia cenocepacia isolates were assessed. The B. cenocepacia environmental isolates (n=35) survived in the hypodermic air pouch but did not invade across the peritoneal epithelial layer during a 72-h infection. For all 35 strains, when the molar ratio of C14:0 3-OH FA to C16:0 3-OH FA in the air pouch fluid wash samples was between 1.4 and 2.5, the concentrations of C14:0 3-OH FA were correlated with the endotoxin unit levels. However, both surrogate markers exhibited different correlations to the inflammatory response. The linear regression coefficient was 0.4234 for C14:0 3-OH FA concentrations vs. NO productions, 0.223 for endotoxin unit levels vs. NO productions, 0.5008 for C14:0 3-OH FA concentrations vs. TNF-alpha productions and 0.2869 for endotoxin unit levels vs. TNF-alpha productions. Therefore, C14:0 3-OH FA concentrations, rather than endotoxin unit levels, acted as an immunostimulatory indicator for LPS in the B. cenocepacia isolates.