Comparative Secretome Analyses of Mycoplasma bovis Virulent and Attenuated Strains Revealed MbovP0145 as a Promising Diagnostic Biomarker

Mycoplasmas are successful pathogens both in humans as well as in animals. In cattle, Mycoplasma bovis (M. bovis) is known to be responsible for serious health complications, including pneumonia, mastitis, and arthritis. However, M. bovis pathogenesis remains unclear. Secreted proteins of M. bovis could influence infection and modify host defense signaling pathways after they enter their extracellular space in the host micro-environment. Therefore, this study was aimed to compare the secretomes of M. bovis HB0801 virulent (P1) and attenuated (P150) strains and identify potential pathogenesis-related secreted proteins and biomarkers. The cells of P1 and P150 strains were grown in pleuropneumonia-like organism medium to log phase and then transferred to phosphate-buffered saline for 2 h. Then, the supernatant was analyzed by using label-free quantitative proteomics, and 477 potential secreted proteins were identified. Combined with the bioinformatics prediction, we found that 178 proteins were commonly secreted by the P1 and P150 strains, and 49 of them were encoded by mycoplasmal core genes. Additionally, 79 proteins were found to have a different abundance between the P1 and P150 strains. Among these proteins, 34 were more abundant and uniquely expressed in P1, indicating a possible association with the virulence of M. bovis. Three differentially secreted proteins, MbovP0145, MbovP0725, and MbovP0174, as well as one equally secreted protein, MbovP0481, as positive control and one protein of inner membrane, MbovP0310, as negative control were, respectively, cloned, expressed, and evaluated for antigenicity, subcellular location, and the secretion nature with their mouse antisera by western blotting and colony immunoblotting assay. Among them, MbovP0145 was confirmed to be more secreted by P1 than P150 strain, highly reactive with the antisera from naturally infected and P1 experimentally infected cattle but not with the P150 vaccinated calves, indicating its potential as a diagnostic antigen. In conclusion, these findings may represent the most extensive compilation of potentially secreted proteins in mycoplasma species and the largest number of differentially secreted proteins between the virulent and attenuated M. bovis strains to date and provide new insights into M. bovis pathogenesis and diagnosis.

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Label-Free and Immobilization-Free Immunoassay

Volume 2: Biomedical and Biotechnology Engineering ◽

10.1115/imece2007-42370 ◽

2007 ◽

Author(s):

Island Pinnick ◽

Kieseok Oh ◽

Chen-Ling Chang ◽

Kyong-Hoon Lee ◽

Jae-Hyun Chung

Keyword(s):

Analytical Model ◽

High Sensitivity ◽

Positive Control ◽

Negative Control ◽

Measured Signal ◽

Label Free ◽

Particle Counter ◽

Fluid Resistance

This paper presents an immunoassay capable of detecting an antigen without labeling or immobilization. By measuring a change in fluid resistance, the immunoassay successfully differentiates a positive control from a negative control. The same device can also act as a particle counter due to its high sensitivity. It is capable of detecting differences in concentrations as low as 104 particles per milliliter. An analytical model is developed to analyze the measured signal.

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Novel Secreted Protein of Mycoplasma bovis MbovP280 Induces Macrophage Apoptosis Through CRYAB

Frontiers in Immunology ◽

10.3389/fimmu.2021.619362 ◽

2021 ◽

Vol 12 ◽

Author(s):

Gang Zhao ◽

Xifang Zhu ◽

Hui Zhang ◽

Yingyu Chen ◽

Elise Schieck ◽

...

Keyword(s):

Coiled Coil ◽

Secreted Proteins ◽

Mycoplasma Species ◽

Mycoplasma Bovis ◽

Secreted Protein ◽

Related Factors ◽

Bioinformatic Tools ◽

Coiled Coil Domain ◽

Αb Crystallin ◽

Apoptosis Inducer

Mycoplasma bovis causes important diseases and great losses on feedlots and dairy farms. However, there are only a few measures to control M. bovis-related diseases. As in other mycoplasma species, this is predominantly because the virulence related factors of this pathogen are largely unknown. Therefore, in this study, we aimed to identify novel virulence-related factors among the secreted proteins of M. bovis. Using bioinformatic tools to analyze its secreted proteins, we preliminarily predicted 39 secreted lipoproteins, and then selected 11 of them for confirmation based on SignalP scores >0.6 or SceP scores >0.8 and conserved domains. These 11 genes were cloned after gene modification based on the codon bias of Escherichia coli and expressed. Mouse antiserum to each recombinant protein was developed. A western blotting assay with these antisera confirmed that MbovP280 and MbovP475 are strongly expressed and secreted proteins, but only MbovP280 significantly reduced the viability of bovine macrophages (BoMac). In further experiments, MbovP280 induced the apoptosis of BoMac treated with both live M. bovis and MbovP280 protein. The conserved coiled-coil domain of MbovP280 at amino acids 210–269 is essential for its induction of apoptosis. Further, immunoprecipitation, mass spectrometry, and coimmunoprecipitation assays identified the anti-apoptosis regulator αB-crystallin (CRYAB) as an MbovP280-binding ligand. An αβ-crystallin knockout cell line BoMac-cryab−, Mbov0280-knockout M. bovis strain T9.297, and its complemented M. bovis strain CT9.297 were constructed and the apoptosis of BoMac-cryab− induced by these strains was compared. The results confirmed that CRYAB is critical for MbovP280 function as an apoptosis inducer in BoMac. In conclusion, in this study, we identified MbovP280 as a novel secreted protein of M. bovis that induces the apoptosis of BoMac via its coiled-coil domain and cellular ligand CRYAB. These findings extend our understanding of the virulence mechanism of mycoplasmal species.

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Endophytic Colletotrichum siamense for Biocontrol and Resistance Induction in Guarana Seedlings

International Journal of Microbiology ◽

10.1155/2021/1925226 ◽

2021 ◽

Vol 2021 ◽

pp. 1-8

Author(s):

Luana L. Casas ◽

José O. Pereira ◽

Pedro Q. Costa-Neto ◽

José F. Silva ◽

Lucas N. Almeida ◽

...

Keyword(s):

First Record ◽

Positive Control ◽

Negative Control ◽

Total Proteins ◽

Resistance Induction ◽

Average Temperature ◽

Main Challenge ◽

Pathogenesis Related ◽

Social Importance ◽

Related Proteins

Paullinia cupana var. sorbilis, known in Brazil as guarana plant, is an important plant and a major traditional crop in the State of Amazonas. It is a native Brazilian species of great economic and social importance, particularly in the Amazon region. Anthracnose caused by Colletotrichum spp. is the main challenge for this crop. Therefore, the present study verified whether C. siamense, an endophytic fungus infected with a mycovirus, could protect the seedlings and reduce or eliminate the characteristic symptoms. Total proteins and enzymatic activities of pathogenesis-related proteins (PRPs), including peroxidase (POX), chitinase (CHI), and phenylalanine ammonia lyase (PAL), were quantified. Guarana seedlings of cultivar Maués were sprayed with a C. siamense conidia suspension (5.0 × 103 conidia/mL). After ten days, the seedlings were sprayed with a suspension of the phytopathogen’s conidia (1.0 × 106 conidia/mL). One group of these seedlings received the fungicide indicated for this crop. The fungicide was applied twice with an interval of 15 days between applications. Negative control seedlings did not receive any treatment (except water and fertilization), and positive control seedlings were treated only with the phytopathogen. The experiment was conducted between December 2019 and February 2020 in a greenhouse. The treatments were applied at an average temperature of 25°C and 85% relative humidity. Leaflets were randomly collected from each treatment group at 0, 48, 72, and 96 hours after pathogen inoculation and analyzed for total protein and enzyme production (POX, PAL, and CHI). After 28 days, the percentage of leaf lesions on the seedlings was evaluated. C. siamense inoculation reduced lesions. There were differences in total proteins and PRPs at different timepoints after inoculation, except for CHI activity, among treatments. To the best of our knowledge, this is the first record of resistance induction in guarana plants.

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Aktivitas Antifungi Air Perasan Bawang Merah (Allium ascalonicum L.) Terhadap Candida albicans Secara In Vitro

Jurnal Ilmu Kedokteran ◽

10.26891/jik.v9i2.2015.71-77 ◽

2017 ◽

Vol 9 (2) ◽

pp. 71

Author(s):

Nurhasanah Nurhasanah ◽

Fauzia Andrini ◽

Yulis Hamidy

Keyword(s):

Candida Albicans ◽

Antifungal Activity ◽

Allium Sativum ◽

Fungicidal Activity ◽

Positive Control ◽

Negative Control ◽

Randomized Design ◽

Significant Difference ◽

Completely Randomized Design

Shallot (Allium ascalonicum L.) has been known as traditional medicine. Shallot which has same genus with garlic(Allium sativum L.) contains allicin that is also found in garlic and has been suspected has fungicidal activity toCandida albicans. It is supported by several researches. Therefore, shallot is suspected has antifungal activity too.The aim of this research was to know antifungal activity of shallot’s water extortion againsts Candida albicans invitro. This was a laboratory experimental research which used completely randomized design, with diffusion method.Shallot’s water extortion was devided into three concentrations, there were 50%, 100% and 200%. Ketoconazole 2%was positive control and aquadest was negative control. The result of this research based on analysis of varians(Anova), there was significant difference between several treatments and was confirmed with Duncan New MultipleRange Test (DNMRT) p<0,05, there was significant difference between 100% shallot’s water extortion with othertreatments, but there was no significant difference between 50% shallot’s water extortion with 200% shallot’s. Theconclusion was shallot’s water extortion had antifungal activity againsts Candida albicans with the best concentration100%, but it was lower than ketoconazole 2%.

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The Comparison of Ostecyte in the Pressure area and Tension area on Tooth Movement Because of Hyperbaric Oxygen Therapy

DENTA ◽

10.30649/denta.v12i1.162 ◽

2018 ◽

Vol 12 (1) ◽

pp. 34

Author(s):

Arya Barahmanta ◽

Muhammad Faizal Winaris ◽

Pambudi Raharjo

Keyword(s):

Hyperbaric Oxygen ◽

Oxygen Therapy ◽

Tooth Movement ◽

Bone Remodelling ◽

Orthodontic Tooth Movement ◽

Negative Control Group ◽

Positive Control ◽

Negative Control ◽

Control Group ◽

Pressure Area

Background: Orthodontic tooth movement is a interaction prosess of resorption and deposition of bone remodeling. Orthodontic tooth movement by mechanical strength causes changes in alveolar bone. Osteocyte is an essential cell to respond bone remodelling. Hyperbaric Oxygen Therapy affects production of osteocyte because it can release Reactive Oxygen Species (ROS) and Nitrid Oxide (NO). Purpose: To determine the difference number of osteocyte in pressure and tension area during tooth movement by adjuvant of Hyperbaric Oxygen 2,4 ATA during 7 days starting on day 8 to day 14. Materials and Methods: This research used Completery Randomized Control Group Post Test Only Design. 36 cavia cobaya (male) were divided into 3 groups randomly : the negative control groups, positive control group, and treatment group. Preparat staining used Hematoxylin Eosin (HE) and calculated on microscop 1000x with 20 field of view. Data analyses used one way ANOVA and LSD test then compared each area by using paired T test. Result: The data showed that the treatment group (P=10,67) tension area has the highest number of osteocyte than negative control group (K-=3,67), positive control (K+=7,42). In the pressure area showed that negative control group (K-=5,00) has the highest than positive control group (K+=3,83) and treatment (P=3,25). Conclusion: Therapy HBO 2,4 ATA 7 days starting on day 8 to day 14 is could increase osteocyte in the tissue to stimulate process of bone remodelling.<pre> </pre>Keywords: Hyperbaric Oxygen, Tooth movement, Bone remodeling, Osteocyte Correspondence: Arya Brahmanta, Department of Orthodonty, Faculty of Dentistry, Hang Tuah University, Arif Rahman Hakim 150, Surabaya, Phone 031-5945864, Email: <a href="mailto:[email protected]">arya.brahmanta@hangtuah.ac.id</a>

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UJI AKTIVITAS HEPATOPROTEKTOR KOMBINASI EKSTRAK ETANOL DAUN PANDAN WANGI (Pandanus amaryllifolius roxb) DAN KAYU MANIS (Cinnamomum burmanii) TERHADAP KADAR MDA YANG DIINDUKSI PARASETAMOL

Jurnal Kesehatan Kusuma Husada ◽

10.34035/jk.v12i1.580 ◽

2020 ◽

pp. 68-73

Author(s):

Yuni Asri Mulatsih Agami ◽

Eka Wisnu Kusuma

Keyword(s):

Oxidative Stress ◽

Ethanol Extract ◽

Optimal Dose ◽

Experimental Methods ◽

Positive Control ◽

Negative Control ◽

Male Rats ◽

Negative Group ◽

Ic50 Value ◽

Dose Combination

Kasus penyakit hati semakin meningkat seiring penggunaan senyawa hepatotoksin salah satunya karena penggunaan parasetamol dengan dosis berlebih. Hal tersebut dapat meningkatkan produksi radikal bebas sehingga memicu terjadinya stress oksidatif yang dapat menimbulkan kerusakan jaringan yang ditandai dengan peningkatan kadar Malondialdehyde (MDA). Stress oksidatif dapat diatasi dengan antioksidan dari berbagai tanaman. Kulit kayu manis memiliki aktivitas antioksidan dengan nilai IC50 53ppm dan daun pandan wangi 39,7% Penelitian ini bertujuan untuk mengetahui aktivitas kombinasi ekstrak etanol daun pandan wangi dan kayu manis dalam menurunkan kadar MDA. tikus yang diinduksi parasetamol. Penelitian menggunakan metode eksperimental, dilakukan selama 9 hari dengan 30 ekor tikus jantan dibagi menjadi 6 Kelompok, yaitu: Normal diberi aquadest, Kontrol Positif diberi silimarin 100 mg/kgBB, Kontrol Negatif diberi CMC-Na 0,05%, serta 3 kelompok lainnya diberi kombinasi ekstrak daun pandan wangi:kayu manis berturut-turut dosis I (25:75), dosis II (50:50), dosis III (75:25). Semua kelompok diinduksi parasetamol 2,5 g/kgBB pada hari ke-7 setelah 30 menit perlakuan, kecuali kelompok normal. Pada hari ke 9 dilakukan pengukuran kadar MDA dengan metode TBARs menggunakan spektrofotometri. Pemberian kombinasi ekstrak etanol daun pandan wangi dan kayu manis dapat menurunkan kadar MDA dengan kombinasi dosis yang paling optimal adalah 75:25 berdasarkan statistik dengan nilai signifikan 0,000<0,05 dibandingkan dengan kelompok negatif. Cases of liver disease have increased with the use of hepatotoxin compounds, one of which is due to the use of paracetamol with excessive doses. This can increase the production of free radicals so that it triggers oxidative stress which can cause tissue damage which is characterized by increased levels of Malondialdehyde (MDA). Oxidative stress can be overcome with antioxidants from various plants. Cinnamomum burmanii has antioxidant activity with IC50 value of 53ppm and Pandanus amarrylifolius 39.7%. This study aims to determine the combined activity of ethanol extract of Pandanus amarrylifolius and Cinnamomum burmanii in reducing MDA levels. Paracetamol-induced rats. Research using experimental methods, conducted for 9 days with 30 male rats divided into 6 groups, namely: Normal given aquadest, Positive Control were given silimarin 100 mg / kgBB, Negative Control was given CMC-Na 0.05%, and 3 other groups were given a combination of Pandanus amarrylifolius extract: Cinnamomum burmanii dose I (25:75), dose II (50:50), dose III (75:25). All groups induced paracetamol 2.5 g / kgBB on the 7th day after 30 minutes of treatment, except the normal group. On the 9th day MDA levels were measured using the TBARs method using spectrophotometry. Giving a combination of Pandanus amarrylifolius and Cinnamomum burmanii ethanol extract can reduce MDA levels with the most optimal dose combination is 75:25 based on statistics with a significant value of 0,000<0.05 compared with the negative group.

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UJI IMUNOMODULATOR DARI EKSTRAK ETANOL DAUN WARU (HIBISCUS TILIACEUS) DENGAN METODE HIPERSENSITIVITAS TIPE LAMBAT

Jurnal Penelitian Farmasi & Herbal ◽

10.36656/jpfh.v1i2.103 ◽

2019 ◽

Vol 1 (2) ◽

pp. 11-16

Author(s):

Yanna Rotua Sihombing ◽

Debi Dinha Sitepu

Keyword(s):

Treatment Group ◽

Leaf Extract ◽

Positive Control ◽

Negative Control ◽

Delayed Type Hypersensitivity ◽

Hypersensitivity Response ◽

Delayed Type Hypersensitivity Response

Immunomodulator is a compound that can increaase the imuno system. One of the plants that have immunomodulator’s activity is Waru Leaf (Hibiscus tiliaceus). the purpose of this research was to test the effect of immunomodulator by extract of Waru Leaf ethanol on rat male. The activity of immunomodulator was determined by using digital pletysmometer by measuring the differences between the last leg swelling’s volume and the first leg swelling’s volume. The treatment group were divided into 5 groups. Each group consistof 5 rats CMC-Na 0,5% (negative control), Stimuno® 32,5 mg/kgBW (positive control), dose of EEDW 50, 100 and 200 mg/kgBW, and bacteria E.coli as antigen. The results slowed that distribution of EEDW dose 200 mg/kgBW can give the effect of immunostimulant by swelling enthancement compared by CMC-Na 0,5 %. EEDW 200 mg/kgBW that have activity comparable with Stimuno® 32,5 mg/kgBW. Thus, it is concluded that of Waru Leaf extract has immunomodulator effects on delayed-type hypersensitivity response of rat male.

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In vitro and in vivo Assessment of Silver Nanoparticles Against Clostridium botulinum Type A Botulinum

Current Drug Discovery Technologies ◽

10.2174/1570163815666180403163946 ◽

2019 ◽

Vol 16 (1) ◽

pp. 113-119 ◽

Cited By ~ 1

Author(s):

Mohammad Aminianfar ◽

Siavash Parvardeh ◽

Mohsen Soleimani

Keyword(s):

Silver Nanoparticles ◽

Botulinum Toxin ◽

Clostridium Botulinum ◽

Lethal Dose ◽

Type A ◽

Positive Control ◽

Negative Control ◽

Laboratory Animals ◽

Control Group ◽

Nano Silver

Background: Clostridium botulinum causes botulism, a serious paralytic illness that results from the ingestion of a botulinum toxin. Because silver nanoparticle products exhibit strong antimicrobial activity, applications for silver nanoparticles in healthcare have expanded. Therefore, the objective of the current study was to assess a therapeutic strategy for the treatment of botulism toxicity using silver nanoparticles. Methods: A preliminary test was conducted using doses that produce illness in laboratory animals to determine the absolute lethal dose (LD100) of botulinum toxin type A (BoNT/A) in mice. Next, the test animals were divided into six groups containing six mice each. Groups I, II and III were the negative control (botulinum toxin only), positive control-1 (nano-silver only) and positive control-2 (no treatment), respectively. The remaining groups were allocated to the toxin that was supplemented with three nano-silver treatments. Results: The mortality rates of mice caused by BoNT/A significantly reduced in the treatment groups with different doses and injection intervals of nano-silver when compared to the negative control group. BoNT/A toxicity induced by intraperitoneal injection of the toxin of Clostridium botulinum causes rapid death while when coupled with nano-osilver results in delayed death in mice. Conclusion: These results, while open to future improvement, represent a preliminary step towards the satisfactory control of BoNT/A with the use of silver nanoparticles for human protection against this bioterrorism threat. Further study in this area can elucidate the underlying mechanism for detoxifying BoNT/A by silver nanoparticles.

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Efek Ekstrak Etanol Kulit Petai (Parkia speciosa Hassk) Terhadap Penurunan Kadar Glukosa Darah Mencit Jantan

Jurnal Katalisator ◽

10.22216/jk.v3i1.2178 ◽

2018 ◽

Vol 3 (1) ◽

pp. 1

Author(s):

Verawaty Verawaty ◽

Dhea Claudia Novel

Keyword(s):

Blood Glucose ◽

Ethanol Extract ◽

Negative Control Group ◽

Positive Control ◽

Negative Control ◽

Control Group ◽

Male Mice ◽

Glucose Levels ◽

The Third ◽

Blood Glucose Levels

Penelitian ini bertujuan untuk melihat pengaruh pemberian ekstrak etanol kulit petai (Parkia speciosa Hassk) terhadap penurunan kadar glukosa darah mencit jantan yang diinduksi aloksan. Hewan percobaan dibagi atas 5 kelompok diantaranya kelompok kontrol negatif, kelompok kontrol positif,dosis I (280 mg/kgBB mencit), dosis II (560 mg/kg BB mencit), dosis III (840 mg/kg BB mencit). Penelitian dilakukan selama 21 hari. Persentase penurunan kadar glukosa darah mencit jantan setelah diberikan ekstrak etanol kulit petai pada hari ke-21 adalah dosis I (77,52 %) lebih besar dibandingkan dengan dosis II (69,5 %) dan dosis III (73,37 %). Data yang diperoleh dianalisis dengan uji Two Way Anova dengan program SPSS 17. Hasil penelitian ini menunjukkan bahwa pemberian ekstrak etanol kulit petai untuk tiga variasi dosis menyatakan perbedaan yang bermakna secara statistik terhadap penurunan kadar glukosa darah mencit jantan.Petai (Parkia speciosa Hassk) has a compound β-sitosterol and stigmasterol that have efficacy to decreased blood glucose levels. This study aimed to determine the effect of ethanol extract of petai peel for decrease blood glucose levels of male mice induced by alloxan. Experimental animals were divided into 5 groups including negative control group, positive control group, the first dose (280 mg/kg in mice), the second dose (560 mg/kg in mice), the third dose (840 mg/kg in mice). The study was conducted for 21 days. After 21 days, the result found that the percentage of blood glucose levels after the male mice given the ethanol extract of petai peel was, the first dose (77.52%) biger than the second dose (69.5%) and the third dose (73.37%). The data obtained were analyzed by Two Way ANOVA using SPSS 17. The results showed that have signicantly difference between three dose variation of ethanol extract of petai peel in blood glucose levels.

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Resistance to Fracture of Dental Roots Obturated with Different Materials

BioMed Research International ◽

10.1155/2015/591031 ◽

2015 ◽

Vol 2015 ◽

pp. 1-5 ◽

Cited By ~ 6

Author(s):

Berkan Celikten ◽

Ceren Feriha Uzuntas ◽

Kamran Gulsahi

Keyword(s):

Fracture Resistance ◽

Testing Machine ◽

Negative Control Group ◽

Positive Control ◽

Negative Control ◽

Positive Control Group ◽

Control Group ◽

Root Canal Filling ◽

Filling Materials ◽

Group 2

The aim of this study was to compare the vertical fracture resistance of roots obturated with different root canal filling materials and sealers. Crowns of 55 extracted mandibular premolar teeth were removed to provide root lengths of 13 mm. Five roots were saved as negative control group (canals unprepared and unfilled). Fifty root canals were instrumented and then five roots were saved as positive control group (canals prepared but unfilled). The remaining 45 roots were randomly divided into three experimental groups (n=15root/group) and obturated with the following procedures: in group 1, glass ionomer-based sealer and cone (ActiV GP obturation system); in group 2, bioceramic sealer and cone (EndoSequence BC obturation system); and in group 3, roots were filled with bioceramic sealer and cone (Smartpaste bio obturation system). All specimens were tested in a universal testing machine for measuring fracture resistance. For each root, the force at the time of fracture was recorded in Newtons. The statistical analysis was performed by using Kruskal-Wallis and post hoc test. There were no significant differences between the three experimental groups. The fracture values of three experimental and negative control groups were significantly higher than the positive control group. Within the limitations of this study, all materials increased the fracture resistance of instrumented roots.

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