Recent advances in the treatment of pain in endometriosis: A bibliometric analysis of experimental models.

Background and Aim: Treatment of endometriosis involves pain relief which is achieved through the administration of analgesics and non-steroidal anti-inflammatory drugs, with or without the addition of hormone therapy. At present, studies investigating endometriosis pain management using experimental rat models and the use of medications are scarce. Therefore, this study aimed to systematically evaluate research trends and critical points in the field of endometriosis pain management using experimental models. Materials and Methods: A total of 30 publications related to this topic that were published from 2012 to 2022 were retrieved from various databases, including Web of Science, Scopus, PubMed, Embase, and CINAHL, using appropriate English keywords. The quality of the publications was evaluated using impact metrics, productivity, term density mapping, and author network. Results: The average publication rate was three articles per year, reaching its peak in 2021 at five articles per year. The United States and China were found to be the most productive countries, with 12 and 10 publications per year, respectively. The field of medicine (37.0%) was the most abundant, although the H-index was relatively low (13:13). Term density mapping involved the search of 542 keywords, of which 35 were selected, with only 8 exhibiting significant density. Conclusion: In the past decade, there has been a shortage of publications that have addressed pain in endometriosis in experimental models and, within this context the majority of the production and publication in this field has been performed by the United States and China. After performing this bibliometric review, it can be inferred that more research is required in this field, to develop new approaches and treatments for endometriotic pain.

Bixa orellana ethyl acetate fraction and its isolated compound ellagic acid attenuate the progression of MIA-induced osteoarthritis in rat knees.

Osteoarthritis (OA) is a pathology that is characterized by progressive erosion of articular cartilage. In this context, medicinal plants have become relevant tools regarding their potential role in the prevention and treatment of OA, being safe and effective. The aim of this work was investigate the therapeutic efficacy of the ethyl acetate fraction of Bixa orellana leaves (BoEA) and ellagic acid (ElAc) for the therapeutic treatment of OA induced by monosodium iodoacetate (MIA) in rats. The plant material was extracted via maceration with 70 % hydroalcoholic solvent (BoHE). The ethyl acetate (BoEA) fraction was by solvents in increasing order of polarity. The ElAc was identified and isolated in BoEA using high performance liquid chromatography (HPLC-DAD) and analytical curve. The OA was induced using MIA in the right knee at the knee joint. Doses of BoEA and ElAc were administered daily (every 24 h, orally) at concentrations of 50, 100 and 50 mg/kg, respectively, for 28 days after induced OA. We evaluated the animals through clinical and radiological examinations every 7 days and, on the 29th day, the animals were euthanized, the joints being removed for histopathological analysis and the serum for cytokine analysis. BoEA and ElAc compounds reduced inflammation and nociception in OA and were as effective as indomethacin in clinical parameters of joint discomfort and allodynia in rats, in addition to showing improvements in radiological and histopathological images, acting on the progress of cartilage deterioration, proving properties related to anti-inflammatory and analgesic processes, being important allies for new therapeutic interventions for the treatment of OA.

Bibliometric Review on New Possibilities of Antimycobacterial Agents: Exploring Siderophore Desferrioxamine's Applications as an Antimicrobial Agent.

Mycobacteria cause tuberculosis and other serious diseases. Understanding their mechanisms of resistance to our immune system and exploring novel drugs are critical strategies to combat infections. A bibliometric analysis was performed to identify publication trends and critical research areas in the field of the antimicrobial activity of desferrioxamine. A total of twenty-four publications on the topic, from 2012 to 2023, were retrieved from databases including Web of Science, Scopus, PubMed, and Embase, using specific keywords. The quality of the publications was assessed using impact and productivity metrics, with an average annual publication rate of 2.1 articles. The United States emerged as the most productive country, with medicine (23.4%, 11 publications) and biochemistry, genetics, and molecular biology (21.3%, 10 publications) as the top research fields. The five most cited publications accounted for 672 citations, with a relatively low h-index (11:11). In conclusion, there has been a lack of publications on this topic in the last decade. The United States dominates production and publication in this area, and there appears to be limited exchange of knowledge, ideas, and technology within the field. Therefore, fostering international cooperation through funding is essential to facilitate further research and development of desferrioxamine-related studies.

Antimicrobial effect of quercetin against Streptococcus pneumoniae.

Streptococcus pneumoniae is a bacterium that causes serious infections, including pneumonia. The limited range of available vaccines and the rise of antibiotic-resistant bacteria mean that new treatments are needed. This study looked at the potential of quercetin as an antimicrobial agent against S. pneumoniae in both isolation and in biofilms. The researchers used microdilution tests, checkerboard assays, and death curve assays, as well as in silico and in vitro cytotoxicity evaluations. They found that quercetin at a concentration of 125.0 µg/mL had both inhibitory and bactericidal effects against S. pneumoniae, and these effects were increased when quercetin was combined with ampicillin. Quercetin also reduced the growth of pneumococcal biofilms. In addition, quercetin (absence or in combination with ampicillin) reduced the death time of Tenebrio molitor larvae compared to the infection control. The study also demonstrated that quercetin had low toxicity in both in silico and in vivo assays, suggesting that it could be a promising treatment for infections caused by S. pneumoniae.

Ethyl Acetate Fraction of Bixa orellana and Its Component Ellagic Acid Exert Antibacterial and Anti-Inflammatory Properties against Mycobacterium abscessus subsp. massiliense.

Mycobacterium abscessus subsp. massiliense (Mabs) causes chronic infections, which has led to the need for new antimycobacterial agents. In this study, we investigated the antimycobacterial and anti-inflammatory activities of the ethyl acetate fraction of Bixa orellana leaves (BoEA) and ellagic acid (ElAc). In silico analysis predicted that ElAc had low toxicity, was not mutagenic or carcinogenic, and had antimicrobial and anti-inflammatory activities. Apparently, ElAc can interact with COX2 and Dihydrofolate reductase (DHFR) enzymes, which could explain both activities. In vitro analysis showed that BoEA and ElAc exerted antimicrobial activity against Mabs (minimum inhibitory concentration of 1.56, 1.56 mg/mL and bactericidal concentration of 6.25, 3.12 mg/mL, respectively. Clarithromycin showed MIC and MBC of 1 and 6 µg/mL). Treatment with BoEA or ElAc increased survival of Tenebrio molitor larvae after lethal infection with Mabs and reduced carrageenan-induced paw edema in mice, around 40% of edema volume after the fourth hour, similarly to diclofenac. In conclusion, BoEA and ElAc exert antimicrobial effects against Mabs and have anti-inflammatory effects, making them potential sources of antimycobacterial drugs. The biological activities of ElAc may be due to its high binding affinities predicted for COX2 and DHFR enzymes.

Restricted Activation of the NF-κB Pathway in Individuals with Latent Tuberculosis Infection after HIF-1α Blockade.

Tuberculous granuloma formation is mediated by hypoxia-inducible factor 1 alpha (HIF-1α), and is essential for establishing latent tuberculosis infection (LTBI) and its progression to active tuberculosis (TB). Here, we investigated whether HIF-1α expression and adjacent mechanisms were associated with latent or active TB infection. Patients with active TB, individuals with LTBI, and healthy controls were recruited, and the expression of cytokine genes IL15, IL18, TNFA, IL6, HIF1A, and A20 in peripheral blood mononuclear cells (PBMCs) and serum vitamin D (25(OH)D3) levels were evaluated. Additionally, nuclear factor kappa B (NF-κB) and tumor necrosis factor-alpha (TNF-α) levels were analyzed in PBMC lysates and culture supernatants, respectively, after HIF-1α blockade with 2-methoxyestradiol. We observed that IL-15 expression was higher in individuals with LTBI than in patients with active TB, while IL-18 and TNF-α expression was similar between LTBI and TB groups. Additionally, serum 25(OH)D3 levels and expression of IL-6, HIF1A, and A20 were higher in patients with active TB than in individuals with LTBI. Moreover, PBMCs from individuals with LTBI showed decreased NF-κB phosphorylation and increased TNF-α production after HIF-1α blockade. Together, these results suggest that under hypoxic conditions, TNF-α production and NF-κB pathway downregulation are associated with the LTBI phenotype.

Ethyl Acetate Fraction of Punica granatum and Its Galloyl-HHDP-Glucose Compound, Alone or in Combination with Fluconazole, Have Antifungal and Antivirulence Properties against Candida spp.

Candidiasis is the most common fungal infection among immunocompromised patients. Its treatment includes the use of antifungals, which poses limitations such as toxicity and fungal resistance. Plant-derived extracts, such as Punica granatum, have been reported to have antimicrobial activity, but their antifungal effects are still unknown. We aimed to evaluate the antifungal and antiviral potential of the ethyl acetate fraction of P. granatum (PgEA) and its isolated compound galloyl-hexahydroxydiphenoyl-glucose (G-HHDP-G) against Candida spp. In silico analyses predicted the biological activity of G-HHDP-G. The minimum inhibitory concentrations (MIC) of PgEA and G-HHDP-G, and their effects on biofilm formation, preformed biofilms, and phospholipase production were determined. In silico analysis showed that G-HHDP-G has antifungal and hepatoprotective effects. An in vitro assay confirmed the antifungal effects of PgEA and G-HHDP-G, with MIC in the ranges of 31.25-250 µg/mL and 31.25 ≥ 500 µg/mL, respectively. G-HHDP-G and PgEA synergistically worked with fluconazole against planktonic cells. The substances showed antibiofilm action, alone or in combination with fluconazole, and interfered with phospholipase production. The antifungal and antibiofilm actions of PgEA and G-HHDP-G, alone or in combination with fluconazole, in addition to their effects on reducing Candida phospholipase production, identify them as promising candidates for therapeutics.

Compounds isolated from Bixa orellana: evidence-based advances to treat infectious diseases / Compuestos aislados de Bixa orellana: avances basados en la evidencia para tratar enfermedades infecciosas

SUMMARY Bixa orellana L. is a native plant from Brazil, but it is also present in other tropical countries such as Peru, Colombia, Ecuador, Mexico, Indonesia, India and East Africa. It is popularly known as Urucum in Brazil. This review shows the potential of bioactive compounds derived from B. orellana to treat infectious diseases due their antimicrobial and antioxidant properties. This plant is also related as an antiinflammatory agent for treatment of pulmonary diseases, or even as eye drops for redness. Its leaves are used for treatment of snakebite, diarrhea, gonorrhea, hepatitis, gastritis, diuretic, antipyretic, and for skin disease. This popular knowledge has encouraged the identification of bioactive compounds in this plant. Compounds as β-cryptoxanthin, geranylgeraniol, lutein, procyanidin B2, procyanidin B3, ellagi tannin isomer and ellagic acid deoxyhexose have been described. These compounds inhibited pathogenic microorganisms such as bacteria, fungi, protozoan and viruses. In addition, some compounds with anti-inflammatory and antioxidant activities were also described. In this sense, B. orellana is a promising source of compounds that could be applied in antimicrobial therapy. This review work may help in the understanding and incentive of new research for antimicrobial discoveries using different B. orellana compounds.

RESUMEN Bixa orellana L. es una planta nativa de Brasil, pero también está presente en otros países tropicales como Perú, Colombia, Ecuador, México, Indonesia, India y África Oriental. Es conocida popularmente como Urucum en Brasil. Esta revisión expone el potencial de los compuestos bioactivos derivados de B. orellana para tratar enfermedades debido a sus propiedades antimicrobianas y antioxidantes. Esta planta también está relacionada como un agente antiinflamatorio para el tratamiento de enfermedades pulmonares e incluso como gotas para los ojos para el enrojecimiento. Sus hojas se utilizan para el tratamiento de la mordedura de serpiente, diarrea, gonorrea, hepatitis, gastritis, diuréticos, antipiréticos y para enfermedades de la piel. Ese conocimiento popular ha fomentado la identificación de compuestos bioactivos en esa planta. Los compuestos β-criptoxantina, geranilgeraniol, luteína, procianidina B2, procianidina B3, isómero elagitanino y ácido elágico desoxihexosa inhibieron microorganismos patógenos como bacterias, hongos, protozoos y virus. En ese sentido, B. orellana es una fuente prometedora de compuestos que podrían aplicarse en la terapia antimicrobiana. Este trabajo de revisión puede ayudar a comprender e incentivar nuevas investigaciones para los descubrimientos de antimicrobianos que utilizan diferentes compuestos de B. orellana.

High frequency of Chlamydia pneumoniae and risk factors in children with acute respiratory infection.

This study was performed as a contribution for a better understanding of Chlamydia pneumoniae frequency in children with respiratory infections. A total of 416 children were recruited from two clinical centers in Sao Luis, Brazil. Of these patients, 165 children had upper respiratory tract infections (URTI), 150 had community-acquired pneumonia (CAP), and 101 were asymptomatic volunteer children. Clinical and epidemiological data from the participants were recorded. Nasopharyngeal swab samples were collected to extract DNA. C. pneumoniae DNA positivity and copy numbers were obtained by an absolute quantitative real-time PCR method. RESULTS: Positivity for C. pneumoniae DNA was higher in samples from URTI children (38.2%) and from CAP children (18.0%) than in those from the control group (7.9%; p < 0.001). Moreover, C. pneumoniae DNA was denser in children with URTI than in asymptomatic children. Considering the cutoff, the highest value of C. pneumoniae DNA found in asymptomatic children of the 3.98 log10 copies/mL, 8.5% (14/165) of the children with URTI, and 3.3% (5/150) with CAP presented high copy numbers of C. pneumoniae DNA. CONCLUSION: Taken together, these results revealed a high frequency of C. pneumoniae in both children with URTI and CAP.

Schinus terebinthifolia leaf lectin (SteLL) has anti-infective action and modulates the response of Staphylococcus aureus-infected macrophages.

Staphylococcus aureus is recognized as an important pathogen causing a wide spectrum of diseases. Here we examined the antimicrobial effects of the lectin isolated from leaves of Schinus terebinthifolia Raddi (SteLL) against S. aureus using in vitro assays and an infection model based on Galleria mellonella larvae. The actions of SteLL on mice macrophages and S. aureus-infected macrophages were also evaluated. SteLL at 16 µg/mL (8 × MIC) increased cell mass and DNA content of S. aureus in relation to untreated bacteria, suggesting that SteLL impairs cell division. Unlike ciprofloxacin, SteLL did not induce the expression of recA, crucial for DNA repair through SOS response. The antimicrobial action of SteLL was partially inhibited by 50 mM N-acetylglucosamine. SteLL reduced staphyloxathin production and increased ciprofloxacin activity towards S. aureus. This lectin also improved the survival of G. mellonella larvae infected with S. aureus. Furthermore, SteLL induced the release of cytokines (IL-6, IL-10, IL-17A, and TNF-α), nitric oxide and superoxide anion by macrophagens. The lectin improved the bactericidal action of macrophages towards S. aureus; while the expression of IL-17A and IFN-γ was downregulated in infected macrophages. These evidences suggest SteLL as important lead molecule in the development of anti-infective agents against S. aureus.

Corrigendum: Galloyl-Hexahydroxydiphenoyl (HHDP)-Glucose Isolated From Punica granatum L. Leaves Protects Against Lipopolysaccharide (LPS)-Induced Acute Lung Injury in BALB/c Mice.

[This corrects the article DOI: 10.3389/fimmu.2019.01978.].

Galloyl-Hexahydroxydiphenoyl (HHDP)-Glucose Isolated From Punica granatum L. Leaves Protects Against Lipopolysaccharide (LPS)-Induced Acute Lung Injury in BALB/c Mice.

The hydroalcoholic extract and ethyl acetate fraction of Punica granatum leaves have been known to exhibit anti-inflammatory activities. In this study, we investigated the therapeutic effects of galloyl-hexahydroxydiphenoyl (HHDP)-glucose isolated from pomegranate leaves on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. Male BALB/c mice were treated with different doses of galloyl-HHDP-glucose (5, 50, and 100 mg/Kg) or dexamethasone at 5 mg/Kg (per os) 6 h after intra-tracheal instillation of LPS. Vehicle-treated mice were used as controls. Twenty-four hours after LPS challenge, bronchoalveolar lavage fluid (BALF), and lung samples were collected for analyses. They were evaluated by monitoring the expression of NF-κB, JNK, and cytokine genes and proteins, as well as cell migration and lung function. All doses of galloyl-HHDP-glucose inhibited LPS-induced JNK and NF-κB activation. Likewise, the galloyl-HHDP-glucose-treated animals presented reduced expression of the TNF-α, IL-6, and IL-1ß genes in the lungs and reduced TNF-α, IL-6, IL-1ß, and IL-8 protein levels when compared with the vehicle-treated LPS-challenged mice. In addition, the ALI mice treated with galloyl-HHDP-glucose also presented reduced lung inflammatory cell accumulation, especially that of neutrophils, in their BALF and lungs. In addition, galloyl-HHDP-glucose treatment markedly ameliorated the LPS-induced pulmonary mechanism complications and attenuated weight loss. Overall, we showed for the first time that galloyl-HHDP-glucose protects against ALI, and may be useful for treating ALI and other inflammatory disorders.

Anti-biofilm Action of Chenopodium ambrosioides Extract, Cytotoxic Potential and Effects on Acrylic Denture Surface.

Considering the challenge to control Candida-associated denture stomatitis, the search for antifungal substances derived from natural sources has become a trend in the literature. In this study the following effects of Chenopodium ambrosioides extract (CAE) were investigated: action against biofilms of Candida albicans, its cytotoxic potential, and changes caused in acrylic resin. The CAE was characterized by High Performance Liquid Chromatography (HPLC). The susceptibility of C. albicans to CAE was investigated by Minimum Inhibitory Concentration and Minimum Fungicidal Concentration (MIC and MFC) tests. Acrylic resin disks were fabricated, and C. albicans biofilms were developed on these for 48 h. Afterward the disks were immersed for 10 min in: PBS (Negative Control); 1% Sodium Hypochlorite (1% SH, Positive Control) or CAE at MIC or 5xMIC. The biofilms were investigated relative to counts and metabolic activity. The cytotoxic potential in keratinocytes and fibroblasts was verified by MTT test. Change in color and roughness of the acrylic resin was analyzed after 28 days of immersion in CAE. The data were analyzed by the ANOVA considering a 5% level of significance. The main compounds detected by HPLC were kaempferol and quercetin. Both MIC and MFC obtained the value of 0.25 mg/mL. The MIC was sufficient to significantly reduce the counts and activity of the biofilm cells (p < 0.0001), while 5xMIC resulted in almost complete eradication, similar to 1% SH. Keratinocytes and fibroblasts exposed to the MIC and 5xMIC presented cell viability similar to that of the Control Group (p > 0.05). No important changes in acrylic resin color and roughness were detected, even after 28 days. It could be concluded that the immersion of acrylic resin in C. ambrosioides extract in its minimum inhibitory concentration was effective for the reduction of C. albicans biofilms without any evidence of cytotoxic effects or changes in roughness and color of this substrate.

Effect of Terminalia catappa Linn. on Biofilms of Candida albicans and Candida glabrata and on Changes in Color and Roughness of Acrylic Resin.

This study aimed to investigate the effect of the n-butanol fraction of Terminalia catappa Linn., (FBuTC) on biofilm of Candida albicans and Candida glabrata, as well as changes in color and roughness of polymethyl methacrylate resin (PMMA). The susceptibility of C. albicans and C. glabrata to FBuTC was evaluated by means of the Minimum Inhibitory and Minimum Fungicidal Concentration (MIC and MFC). PMMA acrylic resin discs (N= 108) were fabricated. For the susceptibility tests, biofilms of C. albicans and C. glabrata were developed on discs for 48 h and immersed in phosphate-saline buffer solution (PBS), 1% sodium hypochlorite (SH 1%), or FBuTC at MIC, 5xMIC, or 10xMIC. For the color and roughness change tests, the discs were immersed in distilled water, SH 1%, or FBuTC in the concentrations of 0.25 mg/mL, 2.5 mg/mL, or 25.0 mg/mL. After 28 days of incubation, color change was evaluated by spectrophotometry and roughness, by using a profilometer. The biofilms were investigated by one-way ANOVA and, the color and roughness changes (two-way ANOVA and the Tukey test; α=0.05). For both MIC and MFC the value of 0.25 mg/mL of FBuTC was observed for the planktonic cells of C. albicans and C. glabrata. Exposure to FBuTC at 10xMIC had a significant effect on the biofilm of C. albicans, showing a reduction in cell counts when compared with PBS, (p=0.001). For the biofilm of C. glabrata, the MIC was sufficient for significantly reducing the cell count (p<0.001). No important changes in color and roughness of the acrylic resin were observed, even after 28 days, irrespective of the concentration of FBuTC used (p >0.05). It could be concluded that the immersion of acrylic resin for dental prosthesis in FBuTC was effective in reducing the biofilms of C. albicans and C. glabrata without evidence of change in roughness and color of this substrate.

Hydroalcoholic Extract and Ethyl Acetate Fraction of Bixa orellana Leaves Decrease the Inflammatory Response to Mycobacterium abscessus Subsp. massiliense.

The incidence of infections caused by rapidly growing mycobacteria (RGM), especially Mycobacterium abscessus subsp. massiliense (Mabs), is increasing worldwide. Severe infections are associated with abscess formation and strong inflammatory response. This study evaluated the antimicrobial and anti-inflammatory activities of a hydroalcoholic extract (BoHE) and ethyl acetate fraction (BoEA) of Bixa orellana leaves. Antimicrobial activity was evaluated by broth microdilution to determine the minimum inhibitory (MIC) and the minimum bactericidal (MBC) concentrations. Cytotoxicity was evaluated using erythrocytes and RAW 264.7 cells. Nitric oxide (NO) was assayed in stimulated RAW 264.7 cells, and inflammatory cell migration and acute toxicity were evaluated in a Mabs-induced peritonitis mouse model. The compounds present in BoEA were identified by high performance liquid chromatography and mass spectrometry (HPLC-MS). The MIC and MBC values were 2.34 mg/mL and 37.5 mg/mL for BoHE and 0.39 mg/mL and 6.25 mg/mL for BoEA. The extracts did not induce significant toxicity in erythrocytes and RAW 264.7 cells. High levels of NO induced by Mabs were decreased by treatment with both extracts. The anti-inflammatory activity was confirmed in vivo by significant reduction of the cell migration to the peritoneum following BoHE and BoEA pretreatment. Animals treated with BoHE or BoEA did not show signs of acute toxicity in stomach, liver, and kidney. The chemical characterization of BoEA (the most active extract) revealed that kaempferol-3-O-coumaroyl glucose is its major component. The extract of B. orellana may be effective for treating infections caused by Mabs.

Bone substitute made from a Brazilian oyster shell functions as a fast stimulator for bone-forming cells in an animal model.

Despite their demonstrated biocompatibility and osteogenic properties, oyster shells have been reported as a potential alternative to other commonly used materials for bone substitution. This study evaluated whether an experimental bone substitute (EBS) made from a typical oyster shell of Northeastern Brazil (Crassostrea rhizophora) has effects on bone development using an animal model. Oysters were collected from a biologically assisted vivarium, and their inner layer was used for preparing an EBS. Chemical and surface characterization of EBS was performed using Individually Coupled Plasma Optical Emission Spectrometry (ICP-OES) and Scanning Electron Microscope (SEM), respectively. Seventy-two rats were randomly assigned to groups according to the treatment of bone defects created in the submandibular area: Negative Control (-C), Positive Control (+C; Bio-Oss®) and EBS. Euthanasia occurred at 7, 21, 42 and 56 days postoperatively. The bone pieces were stained with hematoxylin and eosin (H&E). The formation of bone tissue was evaluated histologically and histomorphometrically. Data were analyzed through the Kruskal-Wallis test and ANOVA considering a significant level of 5%. The main element found in EBS was calcium (71.68%), and it presented heterogeneity in the particle size and a porosity aspect at SEM analysis. Histological results revealed the absence of inflammatory cells in all groups, being that EBS presented the most accelerated process of bone formation with a statistically significant difference between this group and the +C and -C groups in the 21-day time-point (p < 0.05). After 21 days, the bone formation process was similar between all groups (p > 0.05), showing an immature lamellar bone pattern after 56 days of experimentation (p > 0.05). Within the limitations of this study, it was possible to conclude that EBS presented good biocompatibility and promoted fast stimulation for bone-forming cells in an animal model.

Punica granatum L. Leaf Extract Attenuates Lung Inflammation in Mice with Acute Lung Injury.

The hydroalcoholic extract of Punica granatum (pomegranate) leaves was previously demonstrated to be anti-inflammatory in a rat model of lipopolysaccharide- (LPS-) induced acute peritonitis. Here, we investigated the anti-inflammatory effects of the ethyl acetate fraction obtained from the pomegranate leaf hydroalcoholic extract (EAFPg) on the LPS-induced acute lung injury (ALI) mouse model. Male Swiss mice received either EAFPg at different doses or dexamethasone (per os) prior to LPS intranasal instillation. Vehicle-treated mice were used as controls. Animals were culled at 4 h after LPS challenge, and the bronchoalveolar lavage fluid (BALF) and lung samples were collected for analysis. EAFPg and kaempferol effects on NO and cytokine production by LPS-stimulated RAW 264.7 macrophages were also investigated. Pretreatment with EAFPg (100-300 mg/kg) markedly reduced cell accumulation (specially neutrophils) and collagen deposition in the lungs of ALI mice. The same animals presented with reduced lung and BALF TNF-α and IL-1ß expression in comparison with vehicle controls (p < 0.05). Additionally, incubation with either EAFPg or kaempferol (100 µg/ml) reduced NO production and cytokine gene expression in cultured LPS-treated RAW 264.7 macrophages. Overall, these results demonstrate that the prophylactic treatment with EAFPg attenuates acute lung inflammation. We suggest this fraction may be useful in treating ALI.

Identification of specific antibodies against the Ag85C-MPT51-HspX fusion protein (CMX) for serological screening of tuberculosis in endemic area.

OBJECTIVES: Development of new tools for rapid and accurate diagnosis of tuberculosis (TB) is considered a strategy for controlling the disease. The recombinant CMX fusion protein is composed of immunodominant epitopes of the Ag85C (Rv0129c), MPT51 (Rv3803c) and the entire HspX (Rv2031c) proteins from Mycobacterium tuberculosis H37Rv (Mtb). The aim of this study was to evaluate the applicability of a test using the CMX protein in individuals suspected of TB. METHODS: Indirect ELISA was used to measure serum anti-CMX IgM and IgG in individuals with pulmonary TB. RESULTS: Patients with pulmonary TB had higher titers of IgM (OD = 0.502 ± 0.281) than healthy controls (OD = 0.200 ± 0.125). The cutoff for IgM-ELISA was determined using ROC curve analyzes (AUC = 0.868) with a sensitivity of 80.1% and a specificity of 78.2%. Patients with pulmonary TB also had higher titers of IgG (OD = 0.525 ± 0.391) than healthy controls (OD = 0.215 ± 0.077). The cutoff for IgG-ELISA was determined using ROC curve analyzes (AUC = 0.864) with a sensitivity of 81.7% and a specificity of 74.7%. CONCLUSION: The results suggest that the recombinant protein CMX can be used in a serological test to complement the screening of individuals suspected of having active pulmonary TB.

Streptococcus mutans photoinactivation using a combination of a high potency photopolymerizer and rose bengal.

BACKGROUND: Long treatment periods to reach a substantial inactivation of microorganisms are one of the critical challenges in the photodynamic therapy field. METHODS: Planktonic suspensions of Streptococcus mutans were treated in different groups: presence of rose bengal (RB at 2µM) and light exposure by a new high potency photopolymerizer (L at wavelength=440-480nm; dosimetry=96J/cm(2) - 40s of irradiation; potency density=1200mW/cm(2); dosage=48J) - RB+L+ (PDT), just with dye - RB+L-, just with light - RB-L+ and absence of light and dye RB-L- (control group). Aliquots of each group were transferred to Petri dishes to colony counting (CFU/mL) with the data transformed in log10. The results were submitted to ANOVA and Tukey test at 5%. RESULTS: PDT group presented total eradication of microorganisms showing statistical difference with all the other groups (5.82log reduction≅99%). CONCLUSION: The high potency photopolymerizer in pulsed emission at an extra short irradiation and low concentration of rose bengal could be considered as a progressive alternative to the control of S. mutans suspensions.

Immunogenicity of a fusion protein containing immunodominant epitopes of Ag85C, MPT51, and HspX from Mycobacterium tuberculosis in mice and active TB infection.

Tuberculosis (TB) remains a major global health problem. The only vaccine against tuberculosis, attenuated Mycobacterium bovis Bacillus Calmette-Guerin (BCG), has demonstrated relatively low efficacy and does not provide satisfactory protection against the disease in adults. More effective vaccines and better therapies are urgently needed to reduce the global spread of TB. This study evaluated the immunogenicity of a recombinant M. tuberculosis Ag85C-MPT51-HspX fusion protein (CMX) in mice and individuals with active tuberculosis. BALB/c mice were immunized with the CMX protein liposome-encapsulated with CpG DNA or with CpGDNA liposome-encapsulated, liposome or saline as negative controls. The immunization produced high levels of anti-CMX -specific IgG1 and IgG2a antibodies and induced an increase in the relative and absolute numbers of specific TCD4 IFN-γ(+) and TNF-α(+) cells in the spleen. Sera from a cohort of individuals with active tuberculosis contained higher levels of IgG and IgM that recognized CMX when compared to healthy individuals. In conclusion, this protein was shown to be immunogenic both in mice and humans.