RNAi analysis of Doublesex1 expression in Daphnia pulex Leydig, 1860 (Cladocera)

Crustaceana ◽  
2019 ◽  
Vol 92 (2) ◽  
pp. 137-154 ◽  
Author(s):  
Chong-Yuan Lin ◽  
Meng-Di Liu ◽  
Xiao-Jing Zhu ◽  
Jun-Jia Ning ◽  
Shan-Liang Xu ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Fluorescent Protein ◽  
Protein A ◽  
Daphnia Pulex ◽  
Negative Control Group ◽  
Treated Group ◽  
Negative Control ◽  
Control Group ◽  
Blank Control Group ◽  
Rnai Treatment

Abstract To explore the role of the Doublesex gene (Dsx1) in the switching between reproductive modes in Daphnia pulex Leydig, 1860, we performed gene silencing using RNA interference (RNAi). We also investigated the expression of gfp (Green Fluorescent Protein) by quantitative-PCR, and the expression of Dsx1 mRNA after RNAi by real-time PCR. Dsx1 expression was significantly greater in males than in females, and was down-regulated in experimental groups compared with the control group. The decrease was more significant in females (35% of controls) than males (47% of controls). The gene fragment of Dsx1 amplified by PCR was ligated to the pEASY-Blunt vector to obtain the recombinant plasmid expressed recombinant protein induced by isopropyl-β-D-thiogalactoside (IPTG). After having been purified by Ni-column affinity chromatography, the recombinant protein as antigen was used to immune rabbits. The antiserum was successfully purified by the protein A method to obtain the Dsx1 polyclonal antibody, an IgG concentration of 2.65 mg/ml, titer 240 000. Western blotting showed that RNAi treatment significantly reduced DSX1 protein levels. Whole-mount immunofluorescence analysis revealed that the fluorescence intensity of appendages was significantly lower following RNAi treatment than the negative control group, and comparable with the blank control group, further confirming the decrease in Dsx1 expression after RNAi treatment. The number of offspring in all experimental groups was measured, and was significantly greater in the RNAi-treated group compared with the control group. These results provide a foundation for further work on the molecular basis of switching between the two modes of reproduction in cladocerans. The findings also further increase our understanding of the regulatory effects of Dsx1 in the differing sexes in these crustaceans.

UREA IN CORN SILAGE AS A SUPPLEMENTAL NITROGEN SOURCE FOR LACTATING COWS

1979 ◽  
Vol 59 (2) ◽  
pp. 403-410 ◽  
Author(s):  
F. D. SAUER ◽  
J. D. ERFLE ◽  
S. MAHADEVAN ◽  
J. R. LESSARD
Keyword(s):  
Crude Protein ◽  
Protein A ◽  
Daily Intake ◽  
Negative Control Group ◽  
Ammonia Nitrogen ◽  
Negative Control ◽  
Corn Silage ◽  
Feed Consumption ◽  
Control Group ◽  
Protein Nitrogen

Thirty-two Holstein cows in second or later lactation were randomly allocated to four treatment groups within 7 to 10 wk postpartum. Treatment rations were fed ad libitum as a complete feed and consisted of a negative control group which was fed a corn-oats-barley concentrate-corn silage mixture (40:60) with 9.4% crude protein, a urea silage group fed the same grain concentrate mixed with corn silage that contained 0.6% urea (on a fresh weight basis) to give 12.5% total ration crude protein, a group fed a soybean meal concentrate mixed with corn silage (12.7% crude protein), and a group fed a 3% urea corn-oats-barley concentrate mixed with corn silage to give a 12.8% total ration crude protein. The three groups supplemented with protein or non-protein nitrogen had greater weight gains, feed consumption, milk yields, milk persistencies, rumen ammonia nitrogen concentrations and greater rumen microbial cell populations than the negative control group. The results indicate that urea supports milk production when fed as part of a complete feed. Previous work showed that the same daily intake of urea when fed twice daily as part of the concentrate was ineffective.

scholarly journals Prevention of canine ocular thelaziosis (Thelazia callipaeda) with a combination of milbemycin oxime and afoxolaner (Nexgard Spectra®) in endemic areas in France and Spain

Parasite ◽  
2019 ◽  
Vol 26 ◽  
pp. 1 ◽  
Author(s):  
Wilfried Lebon ◽  
Jacques Guillot ◽  
Maria-Jesús Álvarez ◽  
José Antonio Bazaga ◽  
Marie-Laure Cortes-Dubly ◽  
...  
Keyword(s):  
Clinical History ◽  
Negative Control Group ◽  
Treated Group ◽  
Negative Control ◽  
Control Group ◽  
Morphological Identification ◽  
Thelazia Callipaeda ◽  
Milbemycin Oxime ◽  
The Past ◽  
Endemic Areas

In the past decade, canine thelaziosis due to Thelazia callipaeda has been diagnosed in an increasing number of European countries, with endemic areas being identified. A multi-center field trial was conducted in endemic areas in France and Spain to evaluate the efficacy of monthly administrations of the oral milbemycin oxime/afoxolaner combination (NexGard Spectra®) for the prevention of T. callipaeda infection in at-risk dogs. A total of 79 dogs negative for T. callipaeda and with a clinical history of eyeworm infection in the past two years completed the study. Dogs were randomly allocated either to a negative control group (42 dogs) or to the NexGard Spectra® treated group (37 dogs). All dogs were followed up for a 6-month period and assessed monthly for the presence of nematodes on the eyes and for the signs of ocular thelaziosis (e.g., conjunctivitis, keratitis, and ocular discharge). When the presence of nematodes was confirmed, the conjunctival fornix was flushed with a saline solution for parasite recovery and counting, and the dogs were treated appropriately. Recovered parasites were stored in 70% alcohol for subsequent morphological identification. During the course of the study, 57.1% (24/42) of the control dogs were diagnosed positive for Thelazia infection, which illustrates a high incidence rate of parasite infection. Conversely, no eyeworm was recovered from any of the 37 dogs that received NexGard Spectra®. All parasites sampled were confirmed to be T. callipaeda. This clinical field study demonstrated that monthly administrations of NexGard Spectra® provided 100% preventive efficacy against canine thelaziosis.

scholarly journals Immunomodulatory and Anti-Inflammatory Potentials of Trigona Honey in the Therapy and Prevention against Respiratory Infection in Wistar Rats

2020 ◽  
Vol 11 (3) ◽  
pp. 2955-2962
Author(s):  
Ibrahim Khaled Al-kafaween ◽  
Abu Bakar Mohd Hilmi ◽  
Mohamed M. Soliman
Keyword(s):  
Respiratory Infection ◽  
Wistar Rats ◽  
Serum Levels ◽  
Negative Control Group ◽  
Treated Group ◽  
Positive Control ◽  
Negative Control ◽  
Control Group ◽  
Gene Expressions ◽  
Anti Inflammatory

Trigona honey (TH) is well known for its therapeutic characteristics. To date, the study of Trigona honey as a prophylactic or immune booster prior to the bacterial infection of the invivo model is not well covered. This study aims to investigate anti-inflammatory and immune activities in Wistar rats infected with respiratory infection following with Trigona honey. 25 Wistar rats were assigned to possitive groups, negative control group, positive control group was fed TH (5 g / kg body weight) orally, the untreated group was infected with Staphylococcus aureus to induce respiratory infection, the treated group has been infected with S. aureus followed by treatment with TH at a dose of 1.5 ×108 CFU / mL and the preventive group ingested TH one week before S. aureus infection. Blood was obtained for biochemical analysis. Lung tissues have been collected for molecular examination. The results showed a significant decrease in serum levels of ALT, AST, urea and creatinine in the preventive and treated groups, serum IgG increased significantly (P<0.05) in the preventive and treated groups, IFN-y increased in the preventive group while decreased in the treated group, and IL-8 increased in the treated group while decreased in the preventive group. The mRNA expression of AGP is up-regulated in the positive control, preventive and treated groups. The α2-MG, TNF-α , and mRNA expressions showed lower regulation after administration of TH in preventive and treated groups. The results show the ability of TH to counteract immune and inflammatory changes in serum levels and gene expressions.

scholarly journals Nephroprotective effect of virgin coconut oil in Plasmodium berghei ANKA infected Balb/c mice

2021 ◽  
Vol 5 (2) ◽  
Author(s):  
Syafarinah Nur Hidayah Akil ◽  
Heny Arwati ◽  
Puspa Wardhani ◽  
Priangga Adi Wiratama
Keyword(s):  
Plasmodium Berghei ◽  
Coconut Oil ◽  
Negative Control Group ◽  
Treated Group ◽  
Negative Control ◽  
World Health ◽  
Control Group ◽  
Plasmodium Berghei Anka ◽  
Virgin Coconut Oil ◽  
Nephroprotective Effect

AbstractMalaria is a parasitic infectious disease caused by Plasmodium, which remains a world health problem with an estimated 219 million cases worldwide. In severe malaria infection, several organs of the body can be affected, including the kidneys. One of the pathophysiology associated with the worsening of this disease is oxidative stress. The use of antioxidants is expected to prevent this, and one product that has a high antioxidant content is virgin coconut oil (VCO). This study aimed to analyze the effect of VCO on the kidney in Plasmodium berghei ANKA-infected mice. This study was an in vivo laboratory experimental study with a randomized post-test only control group design using 35 BALB/c mice infected with P. berghei ANKA, weighing 20-30 grams. VCO with the Javara® brand is used with doses of 1, 5, and 10 ml/kg body weight (kgBW)/ day. The parameter assessed were levels of BUN, creatinine, and renal histopathological changes. The administration of VCO on the treated group shows minimal tubular necrosis and glomerulonephritis compared to the negative control group. The BUN and creatinine levels in the treated group were also lower than the negative control group. The results showed that VCO has a nephroprotective effect against P. berghei ANKA infection in mice.Keywords: malaria, kidney, virgin coconut oil

scholarly journals Chlorogenic acid prevents alcohol-induced brain damage in neonatal rat

2017 ◽  
Vol 8 (1) ◽  
Author(s):  
Zikang Guo ◽  
Jiang Li
Keyword(s):  
Oxidative Stress ◽  
Cognitive Function ◽  
Chlorogenic Acid ◽  
Inflammatory Mediators ◽  
Caspase 3 ◽  
Negative Control Group ◽  
Treated Group ◽  
Negative Control ◽  
Neonatal Rat ◽  
Control Group

AbstractThe present investigation evaluates the neuroprotective effect of chlorogenic acid (CA) in alcohol-induced brain damage in neonatal rats. Ethanol (12 % v/v, 5 g/kg) was administered orally in the wistar rat pups on postnatal days (PD) 7-9. Chlorogenic acid (100 and 200 mg/kg, p.o.) was administered continuously from PD 6 to 28. Cognitive function was estimated by Morris water maze (MWM) test. However, activity of acetylcholinesterase, inflammatory mediators, parameters of oxidative stress and activity of caspase-3 enzyme was estimated in the tissue homogenate of cerebral cortex and hippocampus of ethanol-exposed pups. It has been observed that treatment with CA attenuates the altered cognitive function in ethanol-exposed pups. There was a significant decrease in the activity of acetylcholinesterase in the CA treated group compared to the negative control group. However, treatment with CA significantly ameliorates the increased oxidative stress and concentration of inflammatory mediators in the brain tissues of ethanol-exposed pups. Activity of caspase-3 enzyme was also found significantly decreased in the CA treated group compared to the negative control group. The present study concludes that CA attenuates the neuronal damage induced in alcohol exposed neonatal rat by decreasing the apoptosis of neuronal cells.

scholarly journals Effects of Steroidal and Nonsteroidal Drugs on Tooth Movement and Root Resorption in the Rat Molar

2009 ◽  
Vol 79 (4) ◽  
pp. 715-726 ◽  
Author(s):  
Carmen Gonzales ◽  
Hitoshi Hotokezaka ◽  
Ken-Ichiro Matsuo ◽  
Tatsunori Shibazaki ◽  
Joseph H. Yozgatian ◽  
...  
Keyword(s):  
Tooth Movement ◽  
Root Resorption ◽  
Three Dimensional ◽  
Negative Control Group ◽  
Treated Group ◽  
Negative Control ◽  
Control Group ◽  
High Dose ◽  
Experimental Control ◽  
Male Wistar Rats

Abstract Objective: To test the hypothesis that the administration of aspirin, acetaminophen, meloxicam, celecoxib, and prednisolone have no effect on root resorption and tooth movement. Materials and Methods: A mesial force of 50 g was applied to the left maxillary first molars of sixty 10-week-old male Wistar rats using nickel titanium closed coil springs attached to the cervical area of the incisors. The rats were randomly divided into 12 groups of 5 each. High and low doses of aspirin, acetaminophen, meloxicam, celecoxib, and prednisolone were administered via drinking water for 2 weeks. The experimental control group had tooth movement but received no drug. The negative control group received neither tooth movement nor drugs. The amount of tooth movement was measured on digitized lateral cephalometric radiographs. Rats were sacrificed after 2 weeks. Mesial and distal roots (distobuccal and distopalatal) were examined using scanning electron and three-dimensional (3D) scanning laser microscopes. The surface area, depth, volume, and roughness of the root resorption craters were measured. Results: When compared with experimental control rats, only prednisolone- and high-dose celecoxib-treated groups showed significantly less root resorption and less tooth movement. Although low dose celecoxib-treated group significantly decreased the tooth movement, root resorption was similar to the control group. Furthermore, resorption craters showed a smoother surface in the prednisolone-treated rats. Conclusions: The hypothesis was rejected. Administration of prednisolone and high-dose celecoxib reduces root resorption and interferes with tooth movement in rats. Both drugs may interfere in the arachidonic acid cascade depending on dose thresholds.

scholarly journals Evaluation of the Wound Healing Activity of the Crude Extract of Leaves of Acanthus polystachyus Delile (Acanthaceae)

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Wubante Demilew ◽  
Getnet Mequanint Adinew ◽  
Seyfe Asrade
Keyword(s):  
Wound Healing ◽  
Medicinal Plants ◽  
Crude Extract ◽  
Negative Control Group ◽  
Treated Group ◽  
Negative Control ◽  
Control Group ◽  
Wound Model ◽  
Treatment Of Wounds ◽  
Wound Healing Activity

Background. Medicinal plants play indispensable roles to treat various ailments. Acanthus polystachyus is one of the medicinal plants used traditionally for treatment of wounds. However, there were no scientific reports documented so far on the wound healing activities of this plant. Thus, the present study provides a scientific evaluation for the wound healing potential of the crude extract of A. polystachyus leaves. Methods. The crude extraction was carried out using 80% methanol. The crude extract was prepared in 5% (w/w) and 10% (w/w) ointment and evaluated for wound healing activity using excision, infected, and incision wound models in Swiss albino mice. Results. Both 5%  and 10% (w/w) ointments significantly reduced period of epithelialization and increased wound contraction rate and tensile strength compared to the negative control group (P < 0.05). The wound healing activity of 10% (w/w) ointment treated group was greater than 5% (w/w) and nitrofurazone ointment treated groups in S. aureus infected wound model. Conclusion. These results demonstrate that the crude extract of A. polystachyus leaves possesses wound healing activities. This justifies the traditional claimed use of the plant for treating uninfected and infected wounds caused by S. aureus.

scholarly journals Effect of miRNA-200b on the proliferation and apoptosis of cervical cancer cells by targeting RhoA

Open Medicine ◽  
2020 ◽  
Vol 15 (1) ◽  
pp. 1019-1027
Author(s):  
Lijie He ◽  
Jing Wang ◽  
Dandan Chang ◽  
Dandan Lv ◽  
Haina Li ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Cancer Cells ◽  
Negative Control Group ◽  
Negative Control ◽  
Luciferase Reporter ◽  
Control Group ◽  
Rt Pcr ◽  
Blank Control Group ◽  
Cervical Cancer Cells ◽  
Mrna And Protein Expression

AbstractObjectiveThis article aims to investigate the effect of miRNA-200b on the proliferation and apoptosis of cervical cancer cells by targeting RhoA.MethodsHeLa cells of cervical cancer were divided into five groups: blank control group, negative control group (miRNA-200b mimic NC), miRNA-200b mimic group, RhoA-negative control group, and RhoA overexpression group. Cells were collected 48 h after transfection. The expression levels of miRNA-200b were detected by RT-PCR. Target relationship between miRNA-200b and RhoA was verified by the dual-luciferase reporter assay. RhoA mRNA and protein expression were detected by western blot and RT-PCR methods. Flow cytometry was used to detect the apoptosis of cells in each group, and the CCK8 method was used to detect the proliferation of cells in each group. The mRNA and protein expression of Bax and cyclin D1 were detected by RT-PCR and western blot.ResultsThe results of the dual luciferase reporter assay showed that RhoA was the target gene of microRNA 200b. Compared with the blank control group and the miRNA-200b mimic-NC group, the proportion of apoptotic cells increased significantly in the miRNA-200b mimic group, and the proliferation of cells was inhibited (P < 0.05). After overexpression of RhoA, the percentage of apoptotic cells decreased and the ability of cell proliferation increased significantly (P < 0.05).ConclusionmiRNA-200b can inhibit the proliferation and promote the apoptosis of cervical cancer cells by targeting the RhoA gene.

scholarly journals The Use of Aloe vera Gel on Scar Collagen

2021 ◽  
Vol 4 (2) ◽  
pp. 89
Author(s):  
Amania Nur Alam ◽  
Muhammad Jailani ◽  
Siti Hajar
Keyword(s):  
Aloe Vera ◽  
Negative Control Group ◽  
Treated Group ◽  
Negative Control ◽  
Control Group ◽  
Control Group Design ◽  
Collagen Formation ◽  
Aloe Vera Gel ◽  
Post Test ◽  
Male Wistar Rats

Aloe vera have active chemicals substances including saponins, tannins, flavonides, and polyphenols. Saponin takes a role as a disinfectant, so it is normally effective for healing any open wounds, while tannins have ability to be an antiseptic. This study aims to see how the effect of the Aloe vera gel on wound collagen, and too inspect which is the most effective concentration of the Aloe vera gel in the formation of wound collagen in wistar strains (Rattus novergicus) rats. This study uses a post-test only method with control group design. The subjects of this study were 28 male Wistar rats (Rattus novergicus). The subjects were divided into 4 groups consisting of 1 negative control group and 3 treated groups, the control group was not given any treatment, while the treated group was given the Aloe vera gel with each concentrations of 12.5%, 25%, and 50% respectively. The experiment was started by making a wound on the back of the rat which then be followed by applying the gel to the wound for 14 days. The results showed the group with the 50% Aloe vera gel treatment had the best collagen formation, followed by a concentration of 25%, 12.5%, and the control group. Therefore, the used of Aloe vera has given good change in scar collagen, and the best scar collagen can be viewed in the group with 50% concentrate of Aloe vera.

scholarly journals Effect of photobiomodulation on the viability of osteoblasts and fibroblasts submitted to alendronate sodium or zoledronic acid

2019 ◽  
Author(s):  
Mariana Aparecida Brozoski ◽  
Natalia Caroline Aguiar Tartaroti ◽  
Andreia Aparecida Traina ◽  
Maria Cristina Zindel Deboni ◽  
Marcia Martins Marques ◽  
...  
Keyword(s):  
Zoledronic Acid ◽  
Cell Viability ◽  
Cell Types ◽  
Negative Control Group ◽  
Treated Group ◽  
Negative Control ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Contact Mode ◽  
Cultured Fibroblasts

The goal of this study is to evaluate the effect of photobiomodulation therapy (PBMT) on the viability of osteoblasts and cultured fibroblasts in different concentrations of alendronate or zoledronic acid. Two cell lines: osteoblast-like mouse cells (OSTEO 1) and human buccal mucosa fibroblast (FMM1) were used. Cells were submitted to different concentrations of bisphosphonates (1 μM, 10 μM, and 100 μM sodium alendronate and 3 μM, 5 μM and 10 μM zoledronic acid) for 24 hours. Next, the cultures received PBMT. The irradiations were applied with a diode laser (InGaAIP, 660 nm, 30 mW, spot 0.028 cm2) in continuous, punctual and contact mode at two energy densities: 5 J/cm2 (4.5 s) or 10 Jcm2 (9s) with 6 hours-intervals. Cell viability was determined by mitochondrial activity assay (MTT) 24 h after the last irradiation. The data were compared by the one way- ANOVA, complemented by the Tukey’s test (p < 0.05). Sodium alendronate at concentrations of 100 μM and 10 μM and zoledronic acid at 10 μM concentration showed higher long-term toxicity. The cellular viability of the PBMT treated group was significantly higher than that of the negative control group. The same occurred with the osteoblasts treated with the highest concentrations of the drug (5 and 10 μM), despite not reaching the cell viability of the positive control group, it presented greater viability than the negative control where the cells were not irradiated. In the groups submitted to zoledronic acid, positive controls presented greater cell viability. We concluded that under the parameters applied in this study, PBMT at an energy density of 5 J/cm2 was able to revert the toxicity of sodium alendronate applied at the higher concentrations in both cell types, whereas zoledronic acid toxicity, regardless of its concentrations, was not influenced by PBMT.

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