scholarly journals From the idea to implementation of the in vivo experiment. Organizing and planning a new protocol

2017 ◽  
Vol 60 (3) ◽  
pp. 250
Author(s):  
A. E. PAPALOIS (Α.Ε. ΠΑΠΑΛΟΗΣ)
Keyword(s):  
Histopathological Examination ◽  
Animal Experiments ◽  
Background Information ◽  
Control Treatment ◽  
Control Group ◽  
Actual Performance ◽  
Treatment Groups ◽  
Definition Of ◽  
Key Steps

A brief overview is presented of the key steps involved in designing a research animal experiment, with reference to resources that specifically address each topic of discussion in more detail. After an idea for a research project is conceived, a thorough review of the literature and consultation with experts in that field are pursued to refine the problem statement and to assimilate background information that is necessary for the experimental design phase. Other practical considerations include defining the necessary control groups, randomly assigning animals to control/treatment groups, determining the number of animals needed per group, evaluating the logistics of the actual performance of the animal experiments and identifying the most appropriate statistical analyses and potential collaborators experienced in the area of study. Also important concerns are anaesthesia, trial dosages of molecules-elements, definition of surgical procedures, sham experiments as control group and cost per experiment. Other factors are probable or expected timeframe for completion of experiments, sufficient financing, pathology and final histopathological examination, specific indicators or values necessary for the evaluation of the results and the origin of the research protocol (doctoral thesis, master or research interests). All of these factors are critical to designing an experiment that will generate scientifically valid and reproducible data, which should be considered the ultimate goal of any scientific investigation.

scholarly journals Efficacy of adjuvant intrastromal and combination of intrastromal and intracameral voriconazole in Aspergillus fumigatus-induced moderate fungal keratitis in rabbits

2021 ◽  
Vol 30 (1) ◽  
pp. 13-9
Author(s):  
Dyah Tjintya Sarika ◽  
Melva Louisa ◽  
Anna Rozaliyani ◽  
Evelina ◽  
Made Susiyanti
Keyword(s):  
Aspergillus Fumigatus ◽  
Clinical Response ◽  
Histopathological Examination ◽  
Inflammatory Cells ◽  
Fungal Keratitis ◽  
Control Group ◽  
Corneal Tissue ◽  
Epithelial Defect ◽  
Treatment Groups

BACKGROUND There is no in vivo evidence for the effectiveness of adjuvant intrastromal and combination of intrastromal and intracameral voriconazole (VCZ) for treating Aspergillus fumigatus keratitis. This study aimed to compare the efficacy of both agents against it.  METHODS A randomized, masked, controlled experimental study was conducted on 11 albino New Zealand white rabbits in which moderate fungal keratitis was induced by inoculating spores of A. fumigatus to the cornea. The rabbits were allocated into 3 groups: 50 μg/0.1 ml intrastromal VCZ injection, 50 μg/0.1 ml intrastromal VCZ and intracameral VCZ injections, and topical VCZ (control). The treatment was given 5 days after inoculation. Epithelial defect, infiltrate size, corneal ulcer depth, and hypopyon were evaluated clinically. Histopathological and mycological examinations were also done 14 days after treatment.  RESULTS All rabbits in the adjuvant treatment groups demonstrated a tendency of a better clinical response with decreasing size of epithelial defect (p = 0.679) and infiltrate (p = 0.755) than in the control group. Direct microscopy, corneal culture, and chop corneal tissue culture were still positive in most of the rabbits from all groups. Histopathological examination showed an increase of inflammatory cells after treatment in all groups, especially in rabbits which were inoculated with A. fumigatus spores in both eyes.  CONCLUSIONS An adjuvant combination of intrastromal and intracameral VCZ showed a tendency of better clinical response for A. fumigatus-induced moderate fungal keratitis in rabbits. 

Mechanism Involved in Fortification by Berberine in CDDP-Induced Nephrotoxicity

2020 ◽  
Vol 13 (4) ◽  
pp. 342-352 ◽  
Author(s):  
Vipin K. Verma ◽  
Salma Malik ◽  
Ekta Mutneja ◽  
Anil K. Sahu ◽  
Kumari Rupashi ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Renal Tissue ◽  
Isoquinoline Alkaloid ◽  
Experimental Models ◽  
Beclin 1 ◽  
Control Treatment ◽  
Control Group ◽  
Treatment Groups ◽  
Single Intraperitoneal Injection ◽  
Male Wistar Rats

Background: The activation of Nrf2/HO-1 pathway has been shown to protect against cisplatin- induced nephrotoxicity by reducing oxidative stress. Berberine (Ber), an isoquinoline alkaloid, has demonstrated antioxidant, anti-inflammatory and anti-apoptotic activities in various experimental models. Aim: To check the effect of Ber on cisplatin-induced nephrotoxicity and to explore the involved mechanism. Methods: Adult male Wistar rats were divided into 6 groups: Normal, cisplatin-control, treatment groups and per se group. Normal saline and Ber (20, 40 and 80 mg/kg; p.o.) was administered to rats for 10 days. A single intraperitoneal injection of cisplatin (8 mg/kg) was injected on 7th day to induced nephrotoxicity. On 10th day, rats were sacrificed, the kidney was removed and stored for the estimation of various parameters. Results: As compared to cisplatin-control group, Ber pretreatment improved renal function system and preserved renal architecture. It also diminished oxidative stress by upregulating the expression of Nrf2/HO-1 proteins. In addition, Ber attenuated the cisplatin mediated inflammation and apoptosis. Furthermore, it also reduced the phosphorylation of p38/JNK and PARP/Beclin-1 expression in the kidney. Conclusion: Ber attenuated renal injury by activating Nrf2/HO-1 and inhibiting JNK/p38MAPKs/ PARP/Beclin-1 expression which prevented oxidative stress, inflammation, apoptosis and autophagy in renal tissue.

scholarly journals In vivo evaluation of a recombinant N-acylhomoserine lactonase formulated in a hydrogel using a murine model infected with MDR Pseudomonas aeruginosa clinical isolate, CCASUP2

AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Masarra M. Sakr ◽  
Walid F. Elkhatib ◽  
Khaled M. Aboshanab ◽  
Eman M. Mantawy ◽  
Mahmoud A. Yassien ◽  
...  
Keyword(s):  
Survival Rate ◽  
Murine Model ◽  
Histopathological Examination ◽  
Healing Process ◽  
Bacterial Count ◽  
Treated Group ◽  
Control Group ◽  
In Vivo Evaluation ◽  
Systemic Spread

AbstractFailure in the treatment of P. aeruginosa, due to its broad spectrum of resistance, has been associated with increased patient mortality. One alternative approach for infection control is quorum quenching which was found to decrease virulence of such pathogen. In this study, the efficiency of a recombinant Ahl-1 lactonase formulated as a hydrogel was investigated to control the infection of multidrug resistant (MDR) P. aeruginosa infected burn using a murine model. The recombinant N-acylhomoserine lactonase (Ahl-1) was formulated as a hydrogel. To test its ability to control the infection of MDR P. aeruginosa, a thermal injury model was used. Survival rate, and systemic spread of the infection were evaluated. Histopathological examination of the animal dorsal skin was also done for monitoring the healing and cellular changes at the site of infection. Survival rate in the treated group was 100% relative to 40% in the control group. A decrease of up to 3 logs of bacterial count in the blood samples of the treated animals relative to the control group and a decrease of up to 4 logs and 2.3 logs of bacteria in lung and liver samples, respectively were observed. Histopathological examination revealed more enhanced healing process in the treated group. Accordingly, by promoting healing of infected MDR P. aeruginosa burn and by reducing systemic spread of the infection as well as decreasing mortality rate, Ahl-1 hydrogel application is a promising strategy that can be used to combat and control P. aeruginosa burn infections.

scholarly journals Comparative studies of the anti-thrombotic effects of saffron and HongHua based on network pharmacology

2020 ◽  
Vol 19 (7) ◽  
pp. 1441-1448
Author(s):  
Jinyan Jiang ◽  
Susu Lin ◽  
Qiaoqiao Li ◽  
Shanshan Jiang ◽  
Yingjie Hu ◽  
...  
Keyword(s):  
Animal Experiments ◽  
Blood Stasis ◽  
Erythrocyte Aggregation ◽  
Network Pharmacology ◽  
Control Group ◽  
Clotting Time ◽  
Aggregation Index ◽  
Model Control

Purpose: To investigate the comparative anti-thrombotic effects of saffron and Honghua, and also to explore possible mechanisms in thrombosis based on network pharmacology. Methods: A network pharmacology model was used for bioactive components, targets and pathways for saffron and HongHua via Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), PharmMapper, Genecard, Uniprot and KEGG databases. In animal experiments, 72 rats were randomly divided into 9 groups: normal control group (NC), model control group (MC), crocetin groups (80, 40, 20 mg/kg), hydroxysafflor yellow A(HSYA) groups (80, 40, 20 mg/kg), and aspirin group (40 mg/kg). Using in vitro thrombosis models and an acute blood stasis model in vivo, the anti-thrombotic effects of these treatments on clotting time, hemorheology parameters, Thromboxane B2 (TXB2), plasmin activator inhibitor (PAI), protein C (PC), protein S (PS), and thrombinantithrombin complex (TAT) were determined and comparisons made for saffron and HongHua. Results: Five potential compounds, 16 anti-thrombotic targets and 27 pathways were predicted for saffron, while 22 compounds, 37 disease targets and 35 pathways were found for HongHua (p < 0.05). Pharmacological experiments revealed that crocetin and HSYA had significant effects on thrombus length, thrombus wet/dry mass, whole blood viscosity (WBV), erythrocyte aggregation index (EAI), clotting time and D-dimer for the high and middle groups. Unlike HSYA, crocetin also had significant and dose-dependent effects on PAI, prothrombin fragment 1+2 (F1+2) and PS and had highly significant effects on TXB2 and TAT. Conclusion: This research provides a systematic, comprehensive and comparative analysis of component, target and anti-thrombotic pathways of saffron and HongHua based on network pharmacology, and also shows that saffron has more significant anti-thrombotic effect than HongHua. Keywords: Saffron; HongHua; Network pharmacology; Anti-thrombosis; Network model

scholarly journals DRES-07. TMZ-LEV- IFN COCKTAIL REGIMEN SIGNIFICANTLY INHIBITED THE GROWTH OF GLIOMA IN VIVO

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi72-vi73
Author(s):  
Xiang-rong Ni ◽  
Jing Wang ◽  
Fu-rong Chen ◽  
Hai-ping Cai ◽  
Yan-jiao Yu ◽  
...  
Keyword(s):  
Protein Expression ◽  
Tumor Growth ◽  
Tumor Volume ◽  
Control Group ◽  
First Line ◽  
Killing Effect ◽  
Treatment Groups ◽  
Mgmt Protein ◽  
Tumor Killing

Abstract OBJECTIVE Temozolomide (TMZ), is the first line chemotherapeutic drug for glioma. Previous studies have suggested that interferon (IFN) and levetiracetam (LEV) could respectively reverse the resistance of TMZ by down-regulating MGMT expression. This study, we aim to investigate the therapeutic effect of a cocktail chemotherapy regimen combining TMZ, LEV, IFN in vivo. METHODS Glioma cell lines U251 and SKMG-4 (MGMT protein expression positive), U138 and GSC-1(MGMT protein expression negative) were used for producing xenograft tumors. The xenograft tumors were established by subcutaneously injecting 1×106 glioma cells into female BALB/C nude mice and divided into 5 treatment groups: Control, TMZ, TMZ+IFN, TMZ+LEV, TMZ+LEV+IFN. The treatment with TMZ (50 mg/kg, i.p.), IFN (2×105 IU, s.c.), LEV (150 mg/kg, i.p.) once a day for five consecutive days and xenograft tumors were measured every two days. RESULTS We identified that U138, U251, SKMG-4 tumor growth among TMZ, TMZ+IFN, TMZ+LEV, TMZ+LEV+IFN were all significantly inhibited (P< 0.05), compared with the control. As for U251 and SKMG-4, tumor killing effect of all 4 treatment groups were not different (P > 0.05). In the treatment of mice bearing U138 glioma, the tumor weight of TMZ+LEV+IFN (0.2688±0.1169 g) group was the lowest and significantly lower than that of TMZ+LEV (0.6574±0.08174g, P=0.0261), TMZ+IFN(0.6108±0.07317 g, P=0.0381), and TMZ (0.9054±0.07154 g, P=0.0017) group. Glioma stem cells GSC-1 was highly resistant to TMZ, tumor volume of TMZ group was not different from control group (P >0.05). While compared with TMZ (1.993±0.1274 g) group, in TMZ+IFN (1.506±0.1223g, P=0.0203), TMZ+LEV (1.178±0.1807g, P=0.0042), and TMZ+LEV+IFN (1.049±0.2171 g, P=0.0038) groups, GSC-1 tumor growth were significantly inhibited(P< 0.05). CONCLUSION Our data demonstrate that both IFN and LEV can sensitize TMZ effect on glioma in vivo, even for MGMT(+) tumors, and TMZ-LEV-IFN cocktail regimen seems the best. Key words: glioma, TMZ, LEV, IFN

scholarly journals Taurine Attenuates Carcinogenicity in Ulcerative Colitis-Colorectal Cancer Mouse Model

2020 ◽  
Vol 2020 ◽  
pp. 1-7 ◽  
Author(s):  
Guifeng Wang ◽  
Ning Ma ◽  
Feng He ◽  
Shosuke Kawanishi ◽  
Hatasu Kobayashi ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Drinking Water ◽  
Mouse Model ◽  
Histopathological Examination ◽  
Tumor Formation ◽  
Water Model ◽  
Control Group ◽  
Ki 67

Taurine (2-aminoethane-sulfonic acid) is a type of amino acids and has numerous physiological and therapeutic functions, including anti-inflammation. However, there are few studies on the anticancer action of taurine. Our previous studies have demonstrated that taurine exhibits an apoptosis-inducing effect on human nasopharyngeal carcinoma cells in vitro. In this study, we have investigated whether taurine has an anticancer effect, using azoxymethane (AOM)/sulfate sodium (DSS)- induced mouse model for colon carcinogenesis. All mice, except those in control group, received a single intraperitoneal injection of AOM and DSS in the drinking water for 7 days twice, with 1-week interval. After the first DSS treatment, mice were given distilled water (model group) or taurine in the drinking water (taurine group) ad libitum. No tumor was observed in the control group. Taurine significantly suppressed AOM+DSS-induced tumor formation. Histopathological examination revealed AOM/DSS treatment induced colon cancer in all mice (8/8, 100%), and taurine significantly inhibited the progression of colon cancer (4/9, 44.4%). Taurine significantly attenuated cell proliferation in cancer tissues detected by Ki-67 staining. Taurine significantly increased the levels of an apoptosis marker cleaved caspase-9 and tumor suppressor protein PTEN. This is the first study that demonstrated that taurine significantly reduced carcinogenicity in vivo using AOM/DSS-induced colon cancer mouse model.

scholarly journals The Chemopreventive Effect of Tanacetum Polycephalum Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene Lactone in MCF7 Cells: A Bioassay-Guided Approach

2015 ◽  
Vol 36 (3) ◽  
pp. 988-1003 ◽  
Author(s):  
Hamed Karimian ◽  
Mehran Fadaeinasab ◽  
Soheil Zorofchian Moghadamtousi ◽  
Maryam Hajrezaei ◽  
Maryam Zahedifard ◽  
...  
Keyword(s):  
Sesquiterpene Lactone ◽  
Histopathological Examination ◽  
Cancer Control ◽  
Control Group ◽  
High Dose ◽  
Mcf7 Cells ◽  
Carcinogenic Effect ◽  
Chemopreventive Effect

Background: Tanacetum polycephalum L. Schultz-Bip is a member of the Asteraceae family. This study evaluated the chemopreventive effect of a T. polycephalum hexane extract (TPHE) using in in vivo and in vitro models. Methods and Results: Five groups of rats: normal control, cancer control, TPHE low dose, TPHE high dose and positive control (tamoxifen) were used for the in vivo study. Histopathological examination showed that TPHE significantly suppressed the carcinogenic effect of LA7 tumour cells. The tumour sections from TPHE-treated rats demonstrated significantly reduced expression of Ki67 and PCNA compared to the cancer control group. Using a bioassay-guided approach, the cytotoxic compound of TPHE was identified as a tricyclic sesquiterpene lactone, namely, 8β- hydroxyl- 4β, 15- dihydrozaluzanin C (HDZC). Signs of early and late apoptosis were observed in MCF7 cells treated with HDZC and were attributed to the mitochondrial intrinsic pathway based on the up-regulation of Bax and the down-regulation of Bcl-2. HDZC induced cell cycle arrest in MCF7 cells and increased the expression of p21 and p27 at the mRNA and protein levels. Conclusion: This results of this study substantiate the anticancer effect of TPHE and highlight the involvement of HDZC as one of the contributing compounds that act by initiating mitochondrial-mediated apoptosis.

Long-term hyperprolactinaemia reduces basal but not androgen-stimulated oestradiol production in small antral follicles of the rat ovary

1991 ◽  
Vol 129 (3) ◽  
pp. 357-362 ◽  
Author(s):  
J. A. Jonassen ◽  
S. P. Baker ◽  
A. S. McNeilly
Keyword(s):  
Female Rats ◽  
Control Group ◽  
Aromatase Activity ◽  
Treatment Groups ◽  
Antral Follicles ◽  
Rat Ovary ◽  
Cervical Stimulation ◽  
Oestradiol Production

ABSTRACT Hyperprolactinaemia disrupts fertility in many species, perhaps by inhibiting ovarian follicular steroidogenesis. The present studies measured oestradiol and progesterone secretion from isolated follicles from rats rendered hyperprolactinaemic in one of two ways. Sustained hyperprolactinaemia was induced by transplantation of two donor pituitary grafts under the renal capsule of adult female rats; grafts remained in place for 3 months. Transient hyperprolactinaemia was induced by pseudopregnancy initiated by cervical stimulation. Small antral follicles were isolated from both groups of rats 8–10 days after the previous vaginal oestrous smear and also from a control group of dioestrous female rats. Follicles were incubated for 3 h in the presence or absence of human chorionic gonadotrophin (hCG) or testosterone. Basal and hCG-stimulated oestradiol production were each reduced in follicles from both hyperprolactinaemic groups, relative to follicles from dioestrous control rats. In contrast, in the presence of testosterone, all groups of follicles produced comparable amounts of oestradiol. hCG stimulated comparable progesterone production by follicles from all three treatment groups. Testosterone elicited smaller increases in progesterone accumulation by follicles from all in-vivo groups. Reduced basal and gonadotrophin-stimulated, but not androgen-stimulated, oestradiol accumulation suggests that androgen production rather than aromatase activity in small antral follicles may be impaired by long-term hyperprolactinaemia. Journal of Endocrinology (1991) 129, 357–362

scholarly journals Pengaruh Pemberian Ekstrak Temulawak (Curcuma Xanthorrizha Roxb.) Terhadap Level Nekrosis pada Jaringan Lien Mencit Putih (Mus Musculus L.) Jantan Galur Balb/C yang Diinokulasi Plasmodium berghei anka

2019 ◽  
Vol 16 (2) ◽  
pp. 186
Author(s):  
MUHAMMAD REYHAN ARSYA ◽  
PRAWESTY DIAH UTAMI ◽  
IRMAWATI IRMAWATI
Keyword(s):  
Mus Musculus ◽  
Plasmodium Berghei ◽  
Histopathological Examination ◽  
Positive Control ◽  
Control Group ◽  
Control Group Design ◽  
Mortality And Morbidity ◽  
Treatment Groups ◽  
Post Test ◽  
Curcuma Xanthorrhiza

<p><strong>Abstract</strong></p><p><strong>Background : </strong>Malaria is a disease caused by the <em>Plasmodium</em> parasite and is transmitted by the <em>Anopheles</em> mosquito and is still a health problem in Indonesia due to high mortality and morbidity. One form of a severe complication of malaria in addition to cerebral malaria is a function failure of the spleen. Today, the management of malaria is increasingly limited due to resistance. Therefore, further development is needed to find new innovations in malaria treatment.</p><p><strong>Purpose : </strong>The purpose of this study was to determine the effect of temulawak rhizome extract (<em>Curcuma xanthorhizza</em> Roxb.) On the level of necrosis in the spleen tissue of male BALB / c mice (<em>Mus musculus</em> L.) inoculated with <em>Plasmodium berghei</em> ANKA.</p><p><strong>Methods :</strong>Experimental research used a post-test only control group design that used five groups of mice. One group of mice was left normal while the other four groups were inoculated with <em>Plasmodium berghei</em> ANKA, positive control groups were given aquades and three treatment groups treated with temulawak extract (<em>Curcuma xanthorrhiza</em> Roxb.) With a dose of 150 mg / KgBB, 100 mg / KgBB, and 50 mg / KgBB for four day. On the fifth day an observation of the level of necrosis in the spleen organ of mice to determine the level of necrosis by histopathological examination using a light microscope.</p><p><strong>Conclusion and Result : </strong>The results of this study indicate that the administration of ginger rhizome extract (<em>Curcuma xanthorriza</em> Roxb.) Has an influence on the level of necrosis of male mice (<em>Mus musculus</em> L.) BALB / c inoculated with <em>Plasmodium berghei</em> ANKA α = 0,002 (ρ&lt;0,05), where the administration of temulawak extract can increase necrosis levels compared to the control group . This is probably due to the lack of temulawak extract dosage and lack of observation in this study.</p><p> </p><p><strong>Keywords </strong>: Malaria, curcuma (<em>Curcuma xanthorrhiza</em> Roxb.), Necrosis level, <em>Plasmodium berghei</em> ANKA</p>

scholarly journals Effects of a reusable progesterone device on conception rates and estrus cycle re-synchronization in Nelore cows

2019 ◽  
Vol 40 (6Supl3) ◽  
pp. 3501
Author(s):  
Fábio Luiz Bim Cavalieri ◽  
Pedro Henrique Baeza Burali ◽  
Fábio Morotti ◽  
Marcelo Marcondes Seneda ◽  
Marcia Aparecida Andreazzi ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Animal Breeding ◽  
Conception Rate ◽  
Control Treatment ◽  
Utilization Rate ◽  
Control Group ◽  
Estrus Cycle ◽  
Treatment Groups ◽  
New Device ◽  
Waste Production

The objective of this study was to evaluate the reuse of an intravaginal progesterone (P4) device during diestrus of Nelore cow embryo recipients in order to improve conception rate, accelerate estrus cycle re-synchronization of non-pregnant animals, and reduce the amount of waste generated by animal breeding biotechnologies. Two experiments were performed using 268 multiparous Nelore cows. In experiment 1, all animals were subjected to a timed embryo transfer (TET) procedure, but at the time of embryo transfer, two treatment groups were established: T1 - the control treatment (N = 132) and T2 - animals receiving a second-use CIDR® device for 12 days (N = 136). Experiment 2 was performed on cows that had not remained pregnant after experiment 1 using two groups: G1 - a control group (N = 69) and G2 -re-synchronized cows that received a P4 device for 12 days for the first TET (N = 74). In experiment 1, no significant effect of the P4 treatment was observed on conception rate (T1 = 37.9%, T2 = 39.7%; P = 0.50) and corpus luteum (CL) diameter (T1 = 17.5 ± 3.4 mm, T2 = 18.1 ± 3.4 mm; P = 0.61). In experiment 2, no significant effect of the treatment was observed on conception rate (G1 = 22.2%, G2 = 35.7%; P = 0.24), recipients utilization rate (G1 = 75.4%, G2 = 70.3%; P = 0.86), and CL diameter (G1 = 17.4 ± 3 mm, G2 = 18.1 ± 3.2 mm; P = 0.27). However, the P4 treatment (for re-synchronization) significantly increased the conception rate (G1 = 22.2%, G2 = 35.7%; P = 0.04), which was similar to that in a conventional TET protocol performed with a new device (38.8%). We conclude that reusable intravaginal P4 devices can help accelerate TET protocols, suggesting an alternative application method; furthermore, this protocol may help reduce waste production in assisted animal breeding.

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