Mouse Foot Research Articles

Liposomal Delivery of the RNA Genome of a Live-Attenuated Chikungunya Virus Vaccine Candidate Provides Local, but Not Systemic Protection After One Dose.

Chikungunya virus (CHIKV) is the causative pathogen of chikungunya fever, a mosquito-borne viral disease causing highly debilitating arthralgia that can persist for months and progress to chronic arthritis. Our previous studies have identified the CHIKV live-attenuated vaccine candidate CHIKV-NoLS. Like most live-attenuated vaccines, attenuated replication of CHIKV-NoLS has the potential to limit scalable production. To overcome production limits, as well as other drawbacks of live-attenuated vaccines, we developed an in vivo liposome RNA delivery system to deliver the self-replicating RNA genome of CHIKV-NoLS directly into mice, allowing the recipients' body to produce the live-attenuated vaccine particles. CAF01 liposomes were able to deliver replication-competent CHIKV-NoLS RNA in vitro. Immunodeficient AG129 mice inoculated with liposome-delivered CHIKV-NoLS RNA developed viremia and disease signs representative of this lethal model of CHIKV infection, demonstrating de novo vaccine particle production in vivo. In immunocompetent C57BL/6 mice, liposome-delivered CHIKV-NoLS RNA inoculation was associated with reduced IgM and IgG levels with low antibody CHIKV-neutralizing capacity, compared to vaccination with the original live-attenuated vaccine CHIKV-NoLS. One dose of liposome-delivered CHIKV-NoLS RNA did not provide systemic protection from CHIKV wild-type (WT) challenge but was found to promote an early onset of severe CHIKV-induced footpad swelling. Liposome-delivered CHIKV-NoLS RNA inoculation did, however, provide local protection from CHIKV-WT challenge in the ipsilateral foot after one dose. Results suggest that in the presence of low CHIKV-specific neutralizing antibody levels, local inflammatory responses, likely brought on by liposome adjuvants, have a role in the protection of CHIKV-induced footpad swelling in the ipsilateral foot of mice inoculated with liposome-delivered CHIKV-NoLS RNA. Low IgG and CHIKV-specific neutralizing antibody levels may be responsible for early onset of severe swelling in the feet of CHIKV-WT-challenged mice. These results support previous studies that suggest CHIKV is vulnerable to antibody-mediated enhancement of disease. Further studies using booster regimes aim to demonstrate the potential for liposomes to deliver the self-replicating RNA genome of live-attenuated vaccines and offer a novel immunization strategy.

Entinociceptive effects of Euphorbia helioscopia extract on Balb/c mice

Background and aims: Euphorbia helioscopia has multiple pharmacological activities, such as antibacterial, antiviral, antifungal,anticancer and/or antitumor, allelopathic, anti-allergic and anti-asthmatic, antioxidant, antinociceptive effect. The aim of the study was to evaluate the antinociceptive activities of Euphorbia helioscopia extract in Balb/c mice, as well as the total flavonoids, phenolic contents, and antioxidant activities of the extract. Methods: In this study, 90 Balb/c mice were randomly designated into 9 groups. Group 1 received normal saline, groups 2 to 7 received different doses of the E. helioscopia hydroethanolic extract (i.e., 0.05, 0.1, 0.2, 0.4, 2, & 8 mg/kg, i.p.).In addition, groups 8 and 9 received naloxone (1 mg/kg) and extract (8 mg/kg) plus naloxone (1 mg/kg), respectively (Naloxone was injected 15 minutes after extract administration). Then, pain response was evaluated for 30 minutes after the injection of 20 µL formalin (1.5%) in the plantar surface of the mice foot. Further, the beta-carotene-linoleate method was used for measuring antioxidant capacity. Finally, total phenolic and flavonoid content were measured based on Folin-Ciocalteu colorimetric and aluminum chloride colorimetric methods, respectively. Results: Total phenol and flavonoid content were 49.43 ± 1.8 mg GAE/g dried extract and 30.19 ± 1.96 mg rutin/g dried extract, respectively. Our results showed that during the first 5 minutes (the acute pain step), a significant difference (P<0.05) was observed between the control group and the group which received the E. helioscopia hydroethanolic extract (8 mg/kg). In the next 25 minutes (the chronic pain step), a significant difference (P<0.05) was found between the control group and the group which received 0.1 and 8 mg/kg doses of the extract. Based on the results, naloxone was unable to reverse the antinociceptive effects of the extract and the maximum antioxidant activity of the extract was 1.641 mg/g of rutin equivalent. Conclusion: In general, this study supports the use of the E. helioscopia extract in folk medicine as the analgesic agent and calls for further investigations regarding elucidating its mechanism of action. Eventually, our findings revealed that the extract of E. helioscopia possessed either antinociceptive or anti-oxidative activities.

Ethanolic Extract Oitment of The Graptophylum Pictum L Griff Leaves as Analgesic agent to Male White Rat

<p>Graptophyllum pictum L Griff Leaves in the community are used as medicinal ingredients, one of which is for analgesics. Black pudding leaves contain secondary metabolite compounds that act as suspected analgesics, namely flavonoids. The ethanolic extract of black pudding leaves has been formulated into cream and ointment preparations. The purpose of this study was to determine the effectiveness of analgesic preparations of black pudding leaves (Graptophyllum pictum L. Griff) ethanol extract in white male mice. The effectiveness of analgesics was tested using the hotplate method. The test animals used were 25 white male mice which were divided into 5 groups: negative control (F0), positive control (Ichtiol® ointment), and black pudding leaf F1 (10%), F2 (15%) ethanol extract treatment group, and F3 (20%). Observation of stretching of mice was done after the feet of mice were induced and immediately given treatment. Observation of analgesic percentage in each mouse was carried out every 5 minutes for 60 minutes. The data obtained were processed and analyzed using one-way ANOVA and Duncan test with a 95% confidence level. One-way ANOVA test results showed that there were significant differences between treatment groups (0.003 <0.05). The results of the average percentage of analgesic effectiveness of formula 2 have a good effect that is equal to 77.969%. Duncan's test results showed that the group that gave the best analgesic effectiveness was formula 2 (15%) comparable to positive control.</p><p><strong>Keywords: Graptophyllum pictum, Analgesics, ointment, Hotplate.</strong><strong></strong></p>

Analgesic effect of Fengshi Qutong Capsules on mice with chronic inflammatory pain caused by complete Freund's adjuvant

The aim was to observe the analgesic effect of Fengshi Qutong Capsules(FSQTC) on chronic inflammatory pain in mice, and investigate its effect on p-ERK/COX-2 signal molecular activity. A model of chronic inflammatory pain was induced in mice by complete Freund's adjuvant(CFA). The mice were divided into normal control group, model group, model+FSQTC 0.3, 0.6 and 1.2 g·kg~(-1 )groups, model+positive control drug ibuprofen(IBP, 0.34 mg·kg~(-1)·d~(-1)) group, and normal control+ FSQTC 1.2 g·kg~(-1)group. FSQTC or IBP was given once a day by oral administration. Standard Von Frey fiber was used to evaluate the mechanical pain threshold, and the acetone stimulation was used to induce inflammatory plantar and observe the cold pain reaction scores. The mechanical pain threshold and cold pain reaction scores were observed before administration and 1, 2, 3, 4, 6 h after administration on the first day, as well as 3 h after administration on the 3 rd to 7 th day. The protein levels of PGE_2, COXs-1,2 and p-ERK in the spinal cord of the inflammatory foot and lumbar 4-5 were detected by enzyme-linked immunosorbent assay, Western blot, immunohistochemistry and immunofluorescence. The results showed that the mechanical pain threshold of the model group decreased and the cold pain reaction score increased as compared with the normal group. FSQTC application could dose-dependently increase the mechanical pain threshold and decrease the cold pain reaction score. The effect lasted for 6 h, most significant at 3 h. The effect of ibuprofen was similar to that of the 0.6 g·kg~(-1) dose group. In addition, FSQTC could reduce the abnormally increased protein content of PGE_2, COX-2 and p-ERK in the inflammatory foot and/or spinal cord of the model group, and the effect was most significant in middle and high dose groups. However, it had no effect on COX-1 in the inflammatory foot and spinal cord of mice. The results suggest that FSQTC has ob-vious analgesic effect on chronic inflammatory pain in mice, which may be related to inhibition of p-ERK/COX-2 signaling pathway.

White blood cell labeling with Technetium-99m (99mTc) using red blood cell extracellular vesicles-mimetics

BackgroundExtracellular vesicles, have gained increasing attention for their application in drug delivery. Here, we developed a novel method for radiolabeling WBCs with 99mTc using RBC-derived extracellular vesicles -mimetics (EVMs), and monitored in vivo inflammation tracking of 99mTc-WBC using gamma camera in acute inflammation mouse model. MethodsEngineered EVMs from RBCs were produced by a one-step extrusion method. RBC-EVMs were analyzed by NTA and TEM. Cells were labeled with 99mTc by using 99mTc-RBC-EVMs. Inflammation mice model was prepared and confirmed by 18F-FDG PET/CT. 99mTc-WBCs were injected in mice, and their biodistribution was analyzed by gamma camera. FindingThe radiochemical purity of 99mTc-RBC-EVMs was 100%. The 99mTc-labeling did't affect the size and morphology. The 99mTc in the cytoplasm of RBC-EVMs was successfully confirmed by high angle annular dark field STEM (scanning transmission electron microscope). Cells were successfully labeled with 99mTc using 99mTc-RBC-EVMs, and the counts per minute was increased in dose- and time-dependent manners. The 18F-FDG PET/CT images confirmed establishment of acute inflammation (left mouse foot). 99mTc-WBCs showed higher uptake in the inflamed foot than non-inflamed foot. InterpretationThis novel method for radiolabeling WBCs using RBC-EVMs. 99mTc labeling may be a feasible method to monitor the in vivo biodistribution of cells.

Development of Botulinum Toxin A-Coated Microneedles for Treating Palmar Hyperhidrosis

Hyperhidrosis is a disorder that is characterized by the production of excess amounts of sweat. The botulinum neurotoxin A (BoNT/A) has been used to treat hyperhidrosis through multiple intradermal injections at the site of the condition. However, because of BoNT/A toxicity, it is important to precisely deliver the proper dose of the toxin to the target site. In addition, the use of a conventional hypodermic needle for multiple injections in the palm makes the approach undesirable and painful. Here, we designed a BoNT/A-coated microneedle (BoNT-MN) array and tested its efficacy as a substitute pain-free method to treat hyperhidrosis. BoNT-MNs were prepared by coating polylactic acid microneedles with a BoNT/A formulation and were found to successfully penetrate into a thick skin in vitro. The coating formulations were then tested for their stability at 4, 25, and 37 °C for 24 h. BoNT-MNs were found to be much more stable than BoNT/A in a liquid state. Additionally, we carried out in vivo experiments by treating the right paws of mice with BoNT-MNs and found that the treatment induced a significant reduction in the sweating response in the mouse foot pad. Thus, BoNT/A treatment using microneedles is beneficial and may be used as a more efficient and less painful approach to treat hyperhidrosis.

Anti-inflammatory effect of artemisinin on uric acid-induced NLRP3 inflammasome activation through blocking interaction between NLRP3 and NEK7

ObjectiveArtemisinin is a potent anti-malarial agent that plays a potent role in regulating inflammatory disorders. NEK7 is a major interacting partner with NLRP3 in NLRP3 inflammasome. The aim of this study was to clarify the anti-inflammatory effect of artemisinin on activation of uric acid-induced NLRP3 inflammasome through regulation of NEK7. MethodsHuman macrophage U937 cells treated with lipopolysaccharide (LPS), monosodium urate (MSU) crystals, or artemisinin were used in in vitro study. Intracellular potassium (K+) level was measured in U937 cells treated with and without artemisinin. Expression of target genes or proteins NEK7, NLRP3, ASC, caspase-1, interleukin-1β (IL-1β), and NF-κB signaling molecules was measured. MSU crystal-induced arthritis model was used for in vivo study. ResultsGout patients showed higher NLRP3 and NEK7 mRNA expression, compared to controls. Enhanced expression of NLRP3, caspase-1, and IL-1β was noted in macrophages treated with LPS (10 ng/ml) and MSU crystals (0.1 mg/ml), which was markedly suppressed by treatment with artemisinin (1, 10, and 100 μM). Artemisinin significantly inhibited interaction between NLRP3 and NEK7 in NLRP3 inflammasome activation. Artemisinin (10 and 100 μM) attenuated intracellular K+ efflux in macrophages stimulated with LPS and MSU crystals. Artemisinin suppressed foot and ankle swelling in MSU crystal-induced arthritis mice. ConclusionThis study revealed that artemisinin inhibited activation of NLRP3 inflammasome by suppressing interaction between NEK7 and NLRP3 in uric acid-induced inflammation.

Low-Cost Gait Analysis for Behavioral Phenotyping of Mouse Models of Neuromuscular Disease.

Measurement of animal locomotion is a common behavioral tool used to describe the phenotype of a given disease, injury, or drug model. The low-cost method of gait analysis demonstrated here is a simple but effective measure of gait abnormalities in murine models. Footprints are analyzed by painting a mouse's feet with non-toxic washable paint and allowing the subject to walk through a tunnel on a sheet of paper. The design of the testing tunnel takes advantage of natural mouse behavior and their affinity for small dark places. The stride length, stride width, and toe spread of each mouse is easily measured using a ruler and a pencil. This is a well-established and reliable method, and it generates several metrics that are analogous to digital systems. This approach is sensitive enough to detect changes in stride early in phenotype presentation, and due to its non-invasive approach, it allows for testing of groups across life-span or phenotypic presentation.

Epigallocatechin-3-Gallate Prevents Acute Gout by Suppressing NLRP3 Inflammasome Activation and Mitochondrial DNA Synthesis.

Gout is a chronic inflammatory disease evoked by the deposition of monosodium urate (MSU) crystals in joint tissues. The nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) family pyrin domain containing 3 (NLRP3) inflammasome is responsible for the gout inflammatory symptoms induced by MSU crystals. We investigated whether epigallocatechin-3-gallate (EGCG) suppresses the activation of the NLRP3 inflammasome, thereby effectively preventing gouty inflammation. EGCG blocked MSU crystal-induced production of caspase-1(p10) and interleukin-1β in primary mouse macrophages, indicating its suppressive effect on the NLRP3 inflammasome. In an acute gout mouse model, oral administration of EGCG to mice effectively alleviated gout inflammatory symptoms in mouse foot tissue injected with MSU crystals. The in vivo suppressive effects of EGCG correlated well with the suppression of the NLRP3 inflammasome in mouse foot tissue. EGCG inhibited the de novo synthesis of mitochondrial DNA as well as the production of reactive oxygen species in primary mouse macrophages, contributing to the suppression of the NLRP3 inflammasome. These results show that EGCG suppresses the activation of the NLRP3 inflammasome in macrophages via the blockade of mitochondrial DNA synthesis, contributing to the prevention of gouty inflammation. The inhibitory effects of EGCG on the NLRP3 inflammasome make EGCG a promising therapeutic option for NLRP3-dependent diseases such as gout.

UJI IMUNOSTIMULATOR DARI KOMBINASI EKSTRAK ETANOL HERBA BINARA (ARTEMISIA VULGARIS LINN) DAN EKSTRAK ETANOL DAUN PUCUK MERAH (SYZYGIUM OLEANA) DENGAN METODE HIPERSENSITIVITAS TIPE LAMBAT PADA TIKUS JANTAN

Diseases mediated by the immune system are difficult problems to treat such as human immunodeficiency virus (HIV) and other lethal viruses. Infections that occur in normal people are generally brief and rarely leave permanent damage. Treatment of this disease requires an aggressive and innovative approach to the development of new treatments so that it requires the role of immunomodulators to improve the immune system. A substance that acts as an enhancer or immune enhancer can be obtained by using herbs that are efficacious as immunostimulants. One of the herbs used is herbal binara (Artemisia vulgaris L) which has been studied as a potential immunomodulator with high antioxidant activity. Previous research also stated that red shoots (Syzygium oleana) were studied as potential immunomodulators with high antioxidant activity. Several other species such as Syzygium samarangense have 16 flavonoida compounds which show pharmacological immunological activity. The purpose of this study was to determine the content of secondary metabolites of ethanol extract of herbal binara (Artemisia vulgaris L.) with red shoots (Syzygium oleana) and to determine the best dose of extract from the ethanol extract of herbal binara (Artemisia vulgaris L.) with red shoots (Syzygium oleana) can reduce the volume of swelling of mouse feet. Examination of the chemical content of secondary metabolites from the ethanol extract of herbal binara (Artemisia vulgaris L.) with red shoots (Syzygium oleana) is carried out by chemical screening and characterization of simplicia and extract. The method used is the slow type hypersensitivity method. In this test the independent variable is the secondary metabolite of ethanol extract of herb binara (Artemisia vulgaris L.) with red shoots (Syzygium oleana) with four concentrations (50, 100, 200 and 400 mg / kgBB). The positive control used by Stimuno dose is 32.5 mg / kgBB

UJI IMUNOSTIMULATOR DARI KOMBINASI EKSTRAK ETANOL HERBA BINARA (ARTEMISIA VULGARIS LINN) DAN EKSTRAK ETANOL DAUN PUCUK MERAH (SYZYGIUM OLEANA) DENGAN METODE HIPERSENSITIVITAS TIPE LAMBAT PADA TIKUS

One of the herbs used is herbal binara (Artemisia vulgaris L) which has been studied as a potential immunomodulator with high antioxidant activity. Previous research also stated that red shoots (Syzygium oleana) were studied as potential immunomodulators with high antioxidant activity. Several other species such as Syzygium samarangense have 16 flavonoida compounds which show pharmacological immunological activity. The purpose of this study was to determine the content of secondary metabolites of ethanol extract of herbal binara (Artemisia vulgaris L.) with red shoots (Syzygium oleana) and to determine the best dose of extract from the ethanol extract of herbal binara (Artemisia vulgaris L.) with red shoots (Syzygium oleana) can reduce the volume of swelling of mouse feet. Examination of the chemical content of secondary metabolites from the ethanol extract of herbal binara (Artemisia vulgaris L.) with red shoots (Syzygium oleana) is carried out by chemical screening and characterization of simplicia and extract. The method used is the slow type hypersensitivity method. In this test the independent variable is the secondary metabolite of ethanol extract of herb binara (Artemisia vulgaris L.) with red shoots (Syzygium oleana) with four concentrations (50, 100, 200 and 400 mg / kgBB). The positive control used by Stimuno dose is 32.5 mg / kgBB.

Cellular and Molecular Level Mechanisms against Electrochemical Cancer Therapy.

Electrochemical treatment (ECT) is a promising new way to induce tumor regression by flowing direct current into the cancer tissue. ECT was applied to different kinds of tumors in clinical studies and showed good results. In addition, basic research has almost not been done in the field of evaluation of efficacy, dose-response, and cytotoxicity. Therefore, the objective is to study the cellular mechanism in the antitumor effect of ECT and to contribute data of basic research of ECT. In the cell-level study, tumor cells (Sarcoma-180, Scc-7, Ehrlich Carcinoma) were studied using ICR mice and C3H mice. In the study group, pH values of control, 10mA × 150secs, 10mA × 300secs, and 10mA × 600secs groups were measured five times each. In histological level studies, ECT was performed on tumors inoculated on the upper part of the right foot of C3H mice. In each group, mice were sacrificed by cervical dislocation 6, 12, and 24 hrs after ECT treatment, and tumors were removed. The excised tumor was fixed in tissue with 10% formalin, and HE staining and apoptosis antibody staining were carried out from the obtained tissue section and observation. In the study at the cellular level, statistically significant differences were observed in all ECT groups in Sarcoma in the tumor growth measurement study compared with the control group. Statistically significant differences were also observed in Scc-7 in all ECT groups compared to the control group. In the intratumoral pH measurement study, there was a statistically significant difference between the anode and the cathode in each group compared to the control group. In the examination at the histological level, microscopic observation of a slide stained with apoptosis antibody with a magnification of 400 times showed that 6hrs after ECT it was stronger and then decreased. By performing ECT, a weak current flows in the living body. As a result, changes in tissue pH, generation of gas, etc. occur. In this study, it was also confirmed that the intratumor pH value becomes strongly acidic on the anode side and strongly alkaline on the cathode side. In addition, this study confirmed the occurrence of gas during treatment of ECT. Changes in the pH and the like cause changes in the environment in the cell, denaturation of proteins, apoptosis, and necrosis. In this study, a significant increase in apoptosis was confirmed in each ECT group compared to the control group. Treatment effects by ECT were also observed in tumor growth measurement studies and tumor weight measurement studies. From these research results, ECT is considered to be effective as a tumor treatment method.

Effect of aging on the tendon structure and tendon-associated gene expression in mouse foot flexor tendon.

To evaluate the biological changes in tendons during the aging process, the present study examined the effect of aging on the tendon structure, distribution of collagen types I and III, and expression of tendon-associated genes, using flexor tendons in a mouse model. Histological assessment of the tendon structure and distribution of collagen types I and III were performed, and the expression of tendon-associated genes was evaluated in flexor digitorium longus tendons of young (8 weeks) and aged (78 weeks) female C57BL/6 mice. The results indicated that the Soslowsky score, based on the analysis of cellularity, fibroblastic changes, and collagen fiber orientation and disruption, was significantly increased, or worsened, in the tendons of the aged group compared with those in the young group. Furthermore, in the aged group, the distribution of type I collagen was decreased and the distribution of type III collagen was relatively increased compared with the young group. Finally, the mRNA expression levels of collagen (type I and type III) and tenogenic markers (Mohawk homeobox, tenomodulin and scleraxis BHLH transcription factor) were significantly decreased in the aged group compared with the young group. The present observations demonstrated that the structure of the tendons, distribution of types I and III collagen and the expression of tendon-associated genes were modulated by aging in the flexor tendon, and that these changes may contribute to the degeneration of tendons in tendinopathy.

Immunomodulatory effect of anise (<i>Pimpinella anisum</i>) in BALB/c mice

Purpose: The aim of this work was to investigate the effect of anise decoction consumption on lymphocytes activity, complete blood count (CBC) and nitric oxide (NO) production in BALB/c mice.Methods: BALB/c mice were given anise tea instead of drinking water and the effect on selected immune parameters was analyzed after 1 and 2 weeks of treatment.Results: Cell activity of anise treated mice was significantly higher than control group at week 2 as revealed by mixed lymphocyte reaction (MLR2. The spleen cells of anise treated mice showed a significant higher lymphocyte proliferative response to in vitro challenge with phytohaemagglutinin (PHA).compared to controls at week two of treatment. The increase in mouse foot thickness as indicator of delayed type of hypersensitivity (DTH) was less in anise treated mice compared to control group. Nitric oxide production by peritoneal macrophages in response to activation with lipopolysaccharides (LPS) was reduced by anise treatment after 1 and 2 weeks of treatment andno significant changes in CD4 and CD8 were noticed either at week 1 or 2 of treatment.Conclusion: This study provides preclinical evidence that anise possesses immunomodulatory activity when administered orally in mice and selectively activates cell-mediated immune mechanismsKeywords: Immunomodulatory, Anise, Traditional medicine

Both ipsilateral and contralateral localized vibratory stimulations modulated pain-related sensory thresholds on the foot in mice and humans.

PurposeThis study was aimed to investigate the effect of localized vibration on sensory thresholds in mice and humans using a novel quantitative method.Participants and methodsThe sensory thresholds of 7-week-old male C57BL/6J mice were measured with four sine-wave electrostimulation frequencies (5, 50, 250, and 2,000 Hz) before and after applying 2-minute vibration to the plantar side of the foot in mice. In human participants (16 males and 16 females; mean age, 21.0±0.8 years), the sensory threshold was measured at 50 Hz before and after applying 2-minute and 5-minute vibrations to the dorsal side of the foot.ResultsApplication of a 2-minute vibration at either the ipsilateral or contralateral side modulated the sensory thresholds elicited by a 5- or 50-Hz right electrostimulation in mice. In human participants, application of a 5-minute vibration at either the ipsilateral or contralateral side modulated the sensory threshold elicited by 50-Hz right electrostimulation, but had no effect on local skin temperature. These results suggest that the right side of pain-related Aδ fibers (50 Hz) or C fibers (5 Hz) was modulated by the localized ipsilateral or contralateral side of vibratory stimuli, respectively, in mice and humans.ConclusionThe ability of contralateral vibration to modify the right sensory thresholds suggests possible involvement of the central nervous system in vibratory modulation.

Suppression of NLRP3 inflammasome by oral treatment with sulforaphane alleviates acute gouty inflammation.

The aetiology of gout is closely linked to the deposition of monosodium uric acid (MSU) crystals and the consequent activation of the NOD-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome. In this study, we investigated whether oral administration of an NLRP3 inhibitor would be effective to attenuate the symptoms of gout. The effects of oral administration with sulforaphane (SFN) were examined in two mouse models of acute gout induced by injection of MSU crystals into footpads or air pouch. The production of caspase-1 (p10) and IL-1β was examined by immunoblotting and ELISA as hallmarks of NLRP3 inflammasome activation. Oral administration of SFN attenuated MSU crystal-induced swelling and neutrophil recruitment in a mouse foot acute gout model, correlating with the suppression of the NLRP3 inflammasome activation in foot tissues. Consistently, oral administration of SFN blocked MSU-crystal-induced activation of the NLRP3 inflammasome in a mouse air pouch gout model. SFN suppressed NLRP3 inflammasome activation induced by MSU crystals, adenosine triphosphate and nigericin but not by poly(dA:dT) in primary mouse macrophages, independent of the reactive oxygen species pathway. SFN inhibited ligand-independent activation of the NLRP3 inflammasome, suggesting that SFN may act directly on the NLRP3 inflammasome complex. Oral administration of SFN effectively alleviated acute gouty inflammation by suppression of the NLRP3 inflammasome. Our results provide a novel strategy in which oral treatment with SFN may be beneficial in preventing acute attacks of gout.

Anti-inflammatory effect of Black cincau (Mesona palustris BL) based supplement in male carrageenan-induced mice

This study aims to investigate the potency and optimum dose of Mesona palustris supplement as an anti-inflammatory in carrageenan-induced mice (Musmus culus). In this study, twenty male Balb/C mice were divided into five groups. The treatment groups are including a negative control, positive control, drug control, and two different doses of black cincau based herbal supplement (BCBHS) groups. In this experiment, edema was observed surrounding mice feet, and the relative number of NF-α and CD68 were measured using flow cytometry analysis. The results revealed that both of BCBHS doses were effective to inhibit edema incidence in carrageenan-induced mice. The expression level of TNF-α and CD68 in both of BCBHS doses were lower than the positive control. Further, the tissue recovery was observed with M. palustris supplementation. Our result suggests that both of BCBHS doses have an anti-inflammatory effect in carrageenan-induced mice.

Near-infrared fluorescence imaging in the largely unexplored window of 900-1,000 nm

Near-infrared (NIR) fluorescence imaging has relied on fluorophores that emit in the 700-900 nm NIR-Ia or 1,000-1,700 nm NIR-II window for generating deep-tissue images. Up until now, there have been few fluorophores developed for the 900-1,000 nm NIR-Ib window. This is largely because NIR-Ib light is thought to be strongly absorbed by water.Methods: Here we found that six heptamethine dyes had distinct emission peaks in both the NIR-Ia and NIR-Ib window. We tested the performance of these contrast agents by introducing them into the leaves of the common house plant Epipremnum aureum with early stage anthracnose leaf infections from Khaya senegalensis, as well as injecting them into the hind feet of nude mice and tails of tumour-bearing mice in vivo.Results: Heptamethine dyes yielded superior images of leaf venation, anthracnose infection locations, sentinel lymph nodes, brain tumours and subcutaneous tumours in the NIR-Ib window. We found that NIR-Ib images had markedly enhanced signal-to-background ratio because autofluorescence, scattering and light absorption by biological tissues and water were weaker at longer wavelengths.Conclusion: NIR-Ib fluorescence imaging was a powerful method for studying sentinel lymph nodes, tumours, leaf veins and early anthracnose infection locations in plant leaves. The findings challenge our current view of NIR fluorescence imaging and may have important implications for biomedical research and image-guided cancer surgery.

Epidemiology and Molecular Diagnosis of Leprosy, Taiwan, 2002-2016

The World Conference on Leprosy, held in 2016, reaffirmed the importance of stopping transmission, preventing disability, and detecting cases early in eradicating the disease. In Taiwan, leprosy is rare and the prevalence rate has reached the eradication criteria. Among the 133 confirmed cases in 2002-2016, 79(59.4%) were female, with 61(72.6%) imported, and the proportion of imported female cases was significant higher than that of the indigenous female cases (p ＜ 0.0001). Since M. leprae cannot be cultured in vitro, bacilli should be inoculated in BALB/c mouse's foot pads. For drug resistance testing, quantitative bacilli are inoculated within a foot pad, and the mice were fed with drug-containing food for 30 weeks. After that, the mice were sacrificed, and the amounts of bacteria were calculated. In order to improve the effectiveness of laboratory diagnosis, we establish an algorithm with molecular tests to identify M. leprae and its drug resistance since 2013. During 2013-2016, of the 47 suspected cases, 27(57.4%) cases were analyzed using the molecular tests. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy were 95.2%, 100%, 100%, 85.7% and 96.3%, respectively. The laboratory tests can strengthen the disease control and management.

Relative biological effectiveness (RBE) and distal edge effects of proton radiation on early damage in vivo

Introduction: The aim of the present study was to examine the RBE for early damage in an in vivo mouse model, and the effect of the increased linear energy transfer (LET) towards the distal edge of the spread-out Bragg peak (SOBP).Method: The lower part of the right hind limb of CDF1 mice was irradiated with single fractions of either 6 MV photons, 240 kV photons or scanning beam protons and graded doses were applied. For the proton irradiation, the leg was either placed in the middle of a 30-mm SOBP, or to assess the effect in different positions, irradiated in 4 mm intervals from the middle of the SOBP to behind the distal dose fall-off. Irradiations were performed with the same dose plan at all positions, corresponding to a dose of 31.25 Gy in the middle of the SOBP. Endpoint of the study was early skin damage of the foot, assessed by a mouse foot skin scoring system.Results: The MDD50 values with 95% confidence intervals were 36.1 (34.2–38.1) Gy for protons in the middle of the SOBP for score 3.5. For 6 MV photons, it was 35.9 (34.5–37.5) Gy and 32.6 (30.7–34.7) Gy for 240 kV photons for score 3.5. The corresponding RBE was 1.00 (0.94–1.05), relative to 6 MV photons and 0.9 (0.85–0.97) relative to 240 kV photons. In the mice group positioned at the SOBP distal dose fall-off, 25% of the mice developed early skin damage compared with 0–8% in other groups. LETd,z = 1 was 8.4 keV/μm at the distal dose fall-off and the physical dose delivered was 7% lower than in the central SOBP position, where LETd,z =1 was 3.3 keV/μm.Conclusions: Although there is a need to expand the current study to be able to calculate an exact enhancement ratio, an enhanced biological effect in vivo for early skin damage in the distal edge was demonstrated.