Prostaglandin E2 induces ovulation in prepubertal mice

The objective of this study was to determine the ability of prostaglandin E2 (PGE2) to induce ovulation and expression of PGE2 receptor (EP2 and EP4) and COX genes (COX-1 and COX-2) in the ovary and pituitary of prepubertal mice. The positive control consisted of the application of 5 μg of gonadotropin-releasing hormone (GnRH, n = 29); the negative control applied 0.5 mL of phosphate buffered saline (PBS, n=31); the treatment tested the application of 250 μg of PGE2 (n = 29), making a total of 89 prepubertal mice (BALB/c). Mice were euthanized 14 to 15 h after treatments to detect ovulation and tissue collection. A Chi-square test was used to compare the proportion of animals ovulating. Gene expressions and number of ovulation were analyzed by one-way ANOVA and Tukey’s test was used to compare means among groups. A greater proportion of mice (P < 0.001) ovulated after receiving GnRH (89.7%, 26/29) compared to PGE2 group (58.6%, 17/29). However, the proportion was higher compared to those treated with PBS (0%, 0/31). Ep2 gene expression in the pituitary was > two-fold higher (P < 0.05) in the PGE2 group compared to the PBS and GnRH groups. Further, PGE2 stimulated Cox1 (2.7 fold, P < 0.05) while GnRH stimulated Cox2 expression (6.5 fold, P < 0.05) in the pituitary when compared to the PBS group. In conclusion, our results support the hypothesis that PGE2 can induce ovulation in prepubertal mice with a concomitant increase in Ep2 and Cox1 gene expression in the pituitary gland.

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322 EFFECT OF PLASMIN ON ACROSOME REACTION OF BUFFALO (BUBALUS BUBALIS) SPERMATOZOA IN VITRO

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab322 ◽

2010 ◽

Vol 22 (1) ◽

pp. 317

Author(s):

I. Venditto ◽

E. Mariotti ◽

L. Boccia ◽

M. Rubessa ◽

M. De Blasi ◽

...

Keyword(s):

Acrosome Reaction ◽

Proteolytic Enzymes ◽

Critical Role ◽

Bubalus Bubalis ◽

Positive Control ◽

Negative Control ◽

Chi Square ◽

Chi Square Test ◽

Response Trial

Fertilization is a critical step of the in vitro embryo production (IVEP) technology in buffalo. It is known that proteolytic enzymes are involved in different steps of the fertilization process; among these, a critical role may be played by the plasminogen activator-plasmin system. It has been demonstrated that plasmin, the active enzyme of this system, induces acrosome reaction (AR) in bull spermatozoa (Taitzoglou IA et al. 2003 Andrologia 35, 112-116). The aim of this study was to investigate the effect of plasmin on the ability of buffalo sperm to undergo the AR. Frozen- thawed sperm from 4 buffalo bulls were treated by swim-up and incubated with 0.01 mM heparin for 4 h. At 0, 2, and 4 h, aliquots of spermatozoa were exposed for 10 min to 60 μg mL-1 of lysophosphatidylcholine (LPC), as positive control, and to 0.01 μg mL-1 of plasmin. This concentration was chosen after a preliminary dose-response trial. Another sample from each treatment was incubated with IVF medium (negative control). After 10 min, sperm motility was evaluated and sperm were fixed in 37% formaldehyde and stained with trypan blue-Giemsa for subsequent microscopic examination. The total number of sperm counted, over 3 replicates, was 1269 for the negative control, 1293 for LPC, and 1238 for plasmin, equally distributed among incubation times. Differences among groups were analyzed by chi-square test. After swim-up, acrosomal loss was observed only in 4% of the sperm. The addition of 0.01 μg mL-1 of plasmin for 10 min to buffalo spermatozoa at time 0 significantly (P < 0.01) enhanced (23%) AR compared with the control (7.8%), with the same efficiency of LPC (17.1%). After 2 h of incubation with heparin, both plasmin and LPC increased the AR compared to the control (24.4, 20.1, and 14.0%, respectively; P < 0.01). After 4 h, plasmin gave higher percentages of AR (27.2%) compared to both the control (21.0%; P < 0.05) and LPC (19.2%; P < 0.01). Another interesting result is the improved motility recorded with plasmin compared to both the control and LPC groups at 2 h of incubation (90, 75, and 75%, respectively; P < 0.05) and at 4 h of incubation (75, 60, and 60%, respectively; P < 0.05). Finally, no differences in sperm viability were observed between plasmin and the control, whereas a decreased viability was found when LPC was used at 0 h (96.2, 95.0, and 89.0%, respectively, for plasmin, control, and LPC; P < 0.05), at 2 h (85.0, 87.5, and 77.0%, respectively, for plasmin, control, and LPC; P < 0.01), and at 4 h (85.0, 93.3, and 81.1%, respectively, for plasmin, control, and LPC; P < 0.01). In conclusion, we found that addition of plasmin to capacitated sperm increases the percentage of acrosome-reacted spermatozoa and improves motility. Our results suggest that plasmin may play a role in events surrounding fertilization and suggest to evaluate in further studies whether the addition of plasmin during IVF improves the IVEP efficiency in buffalo.

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A Case-Control Study on the Relationship Between Cholecystectomy and Cholangiocarcinoma in China

10.21203/rs.3.rs-136501/v1 ◽

2020 ◽

Author(s):

Donglie Zhu ◽

Hang Fu ◽

Zelong Yang ◽

Mingzuo Jiang ◽

Yanjie Ren ◽

...

Keyword(s):

Negative Control Group ◽

Positive Control ◽

Negative Control ◽

Control Group ◽

Case Group ◽

Hepatic Hemangioma ◽

Chi Square ◽

Chi Square Test ◽

History Of ◽

The Difference

Abstract Aims: The present study aimed to explore the correlation between cholecystectomy and cholangiocarcinoma, and to provide preliminary clinical basis for precise cholecystectomy in China.Methods: We conducted a retrospective analysis of 9744 patients with cholangiocarcinoma, colon cancer, pancreatic cancer, femoral fracture, and hepatic hemangioma diagnosed in Xijing hospital from August 2008 to August 2018. They were divided into three groups: case group (1749 cases of cholangiocarcinoma), positive control group (3137 cases of colon cancer and 1950 cases of pancreatic cancer), negative control group (1794 cases of femoral fracture and 1114 cases of hepatic hemangioma). We collected the general information (gender, age), past medical history, cholecystectomy history from the patients, and these data were analyzed by chi-square test and logistic regression analysis. Results: The cholecystectomy rate of the case group was significantly higher than that of the positive control group and the negative control group by chi-square test (p<0.025). The cholecystectomy rate and the history of cholecystolithiasis were analyzed by logistic multivariate regression analysis. The OR values of cholecystectomy rate were 1.553 (95%CI: 1.311-1.840) and 3.181 (95%CI: 2.561-3.951), respectively, and the difference was statistically significant (p<0.000). The OR values of the history of cholecystolithiasis were 2.460 (95%CI: 2.093-2.890) and 5.426 (95%CI: 4.325-6.809), respectively, and the difference was statistically significant (p<0.000). In case group, the difference between cholecystectomy and cholecystolithiasis was statistically significant (p<0.000) by chi-square test. Conclusions: In conclusion, cholecystectomy is one of the risk factors of cholangiocarcinoma and the patients who undergo cholecystectomy have a higher risk of cholangiocarcinoma than the control groups. Cholecystectomy should be conducted with caution and the precise surgical treatment of gallbladder diseases is advocated.

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311 MELATONIN PROMOTES IN VITRO SPERM CAPACITATION IN BUFFALO (BUBALUS BUBALIS)

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab311 ◽

2010 ◽

Vol 22 (1) ◽

pp. 311 ◽

Cited By ~ 5

Author(s):

S. Di Francesco ◽

E. Mariotti ◽

M. Tsantarliotou ◽

A. Sattar ◽

I. Venditto ◽

...

Keyword(s):

Acrosome Reaction ◽

Optimal Dose ◽

Bubalus Bubalis ◽

Positive Control ◽

Negative Control ◽

Sperm Capacitation ◽

Chi Square ◽

Melatonin Administration ◽

Chi Square Test

Melatonin, the main hormone secreted by the pineal gland, plays many roles in reproduction. In ram spermatozoa, melatonin administration increases plasminogen activator activity (Tsantarliotou MP et al. 2007 Theriogenology 69, 458-465), known to be involved in sperm capacitation and acrosome reaction (Taitzoglou IA et al. 1996 Mol. Androl. 8, 187-197). The aim of this study was to evaluate the efficiency of melatonin to induce buffalo in vitro sperm capacitation, indirectly assessed by estimating the capability of spermatozoa to acrosome-react. Frozen-thawed semen from 4 different bulls was pooled and treated by swim-up in order to select only the motile population. Spermatozoa (n = 829) were assessed immediately after swim- up separation, to evaluate the incidence of acrosomal loss in non-treated cells (time 0). The remaining spermatozoa were incubated in the absence of capacitating agents (negative control; n = 513), in the presence of 0.01 mM heparin (positive control; n = 775), 10 μM melatonin (n = 684), 100 μM melatonin (n = 751), and 1 mM melatonin (n = 650), for 2 h. Sperm were then exposed for 15 min to 60 μg mL-1 of lysophosphatidylcholine, an agent known to induce acrosome reaction (AR) only on capacitated spermatozoa. Trypan blue was used first to differentiate live from dead spermatozoa and the dried smears were then fixed in 37% formaldehyde and stained with Giemsa for acrosome evaluation by microscopic examination. The percentage of acrosome-reacted spermatozoa in each group was used to assess the efficiency of capacitation under different incubation conditions. The experiment was repeated 4 times. Differences among groups were analyzed by chi-square test. Acrosomal loss was observed only in 2.1% of the sperm population at time 0. Interestingly, sperm treatment with both heparin and the different concentrations of melatonin resulted in a significantly higher incidence of AR compared to the negative control (24.4, 20.5, 20.0, 23.6 v. 8.0% for the positive control, 10 μM melatonin, 100 μM melatonin, 1 mM melatonin, and the negative control, respectively; P < 0.01). These results demonstrated that melatonin determines capacitation of buffalo spermatozoa in vitro. Furthermore, the effect of melatonin was comparable to that of heparin, that is, the capacitating agent currently used in the IVF system. The capacitating effect was observed at all the tested concentrations, and viability was not affected. This suggests to extend the range of concentrations to test in future studies, in order to identify the optimal dose. Moreover, considering the seasonality of the species and the great differences in fertility attitude of buffalo bulls, it would be interesting to investigate the capacitation effect of melatonin in relation to both the season and the bull.

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Prevalence of various types of allergens in ocular allergic conditions of patients from Pokhara, Nepal

Nepalese Journal of Ophthalmology ◽

10.3126/nepjoph.v6i1.10759 ◽

2014 ◽

Vol 6 (1) ◽

pp. 6-23 ◽

Cited By ~ 1

Author(s):

Sachet Prabhat Shrestha ◽

Amod K Pokhrel ◽

Pushpa Malla ◽

Srijana Thapa Godar

Keyword(s):

Skin Prick Test ◽

Allergic Conjunctivitis ◽

Allergic Diseases ◽

Positive Control ◽

Negative Control ◽

Test Reaction ◽

Chi Square ◽

Prick Test ◽

Chi Square Test ◽

Skin Prick Tests

Introduction: Ocular allergic conditions are mostly recurrent and the drugs prescribed, especially corticosteroids, have serious side effects. Therefore, when maximal tolerated topical and systemic medications are unable to control allergic conjunctivitis, a skin prick test for allergens should be conducted and patients should be taught to avoid these allergens. Objective: To find out the prevalence of common allergens inciting ocular allergic diseases in Nepal. Subjects and methods: A total of 13,376 skin prick tests were performed on 76 patients suffering from different chronic recurrent ocular allergic conjunctivitis with 176 common allergens (pollens, fungi, insects, dusts, danders, fabrics/ feathers, food, parthenium leaves, tobacco and mite). Buffer saline was used as a negative control while histamine acid phosphate was used as a positive control. Grading of the skin prick test reaction was done by comparison to the histamine positive control. Only markedly positive reactions were considered positive. Relevant data were entered into the excel spreadsheet and analyzed with the Stata-12 commercial package. The association between allergic conditions and socio-demographic, environmental and other co-variates were tested by the chi-square test. Results: The common offenders found in the study were mite (42.11 %) followed by fabrics/ feathers (20.39 %), dusts (18.18 %), pollen (17.05 %), non-juicy food (15.02 %), dander (13.60 %), juicy food (11.64 %) and fungus (9.87 %), and tobacco (6.58 %), parthenium leaves (5.26 %) and insects (3.17 %) were less common offenders. Conclusions: All ocular allergy patients should undergo skin prick tests to find out the allergens causing their allergy and then receive specific immunological treatment (SIT). DOI: http://dx.doi.org/10.3126/nepjoph.v6i1.10759 Nepal J Ophthalmol 2014; 6 (2): 6-23

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145 EFFECT OF RELAXIN ON FERTILIZING ABILITY OF BUFFALO SPERM

Reproduction Fertility and Development ◽

10.1071/rdv26n1ab145 ◽

2014 ◽

Vol 26 (1) ◽

pp. 186 ◽

Cited By ~ 2

Author(s):

A. R. Elkhawagah ◽

V. Longobardi ◽

G. A. Sosa ◽

G. Albero ◽

A. Salzano ◽

...

Keyword(s):

Sperm Motility ◽

Positive Control ◽

Negative Control ◽

Cleavage Rate ◽

Chi Square ◽

Chi Square Test ◽

Live Sperm ◽

Student’S T ◽

Fertilizing Capability

The aim of this work was to evaluate the effect of relaxin, known to improve fertility parameters of frozen-thawed sperm in other species (Miah et al. 2006 J. Reprod. Dev. 52, 773–779; Miah et al. 2007 Anim. Sci. J. 78, 495–502), on buffalo sperm motility, capacitation, and fertilizing capability. Frozen-thawed sperm from 2 bulls (4 replicates each) were separated by Percoll, diluted to a 20 × 106 mL–1 concentration and incubated in TALP medium in the absence of capacitating agents (negative control), in the presence of 10 μg mL–1 of heparin (positive control) and 100 ng mL–1 of relaxin for 2 h. Following incubation, sperm were exposed for 15 min to 60 mg mL–1 of lysophosphatidylcholine, a fusogenic agent known to induce the acrosome reaction only on capacitated sperm. To evaluate acrosome-reacted (AR) live sperm, cells were fixed and stained with Trypan blue-Giemsa (Kovacs and Foote 1992 Biotech. Histochem. 67, 119–124) and evaluated (800 sperm counted/group). Sperm motility was examined by a phase contrast microscope, whereas the fertilizing capability was evaluated by heterologous IVF. Abattoir-derived bovine oocytes (n = 258, 86 per group) were in vitro matured and fertilized according to standard procedures (Rubessa et al. 2011 Theriogenology 76, 1347–1355) with buffalo sperm in the absence of capacitating agents and in the presence of 10 μg mL–1 of heparin and 100 ng mL–1 of relaxin. Twenty hours after IVF, presumptive zygotes were denuded and cultured in SOF for 24 h, when cleavage rate was evaluated and confirmed by fixation with absolute ethanol overnight and staining with 2.5 μg mL–1 of Hoechst 33342 after zona removal by pronase (2 mg mL–1) digestion. The differences in the percentages of AR sperm and cleavage among groups were analysed by a chi square test and those in sperm motility by Student's t-test. Acrosomal loss was observed in 10.8% of the sperm after thawing, which may indicate freezing-induced capacitation, and, hence, this value was detracted from the percentages of AR recorded following incubation. After 2 h of incubation, 100 ng mL–1 of relaxin significantly (P < 0.05) increased the percentages of live AR sperm (P < 0.05) compared with the negative control (31.3 ± 2.2 and 25.8 ± 2.8, respectively), with intermediate results in the positive control (27.0 ± 2.2). Motility was significantly improved (P < 0.05) when sperm were exposed to 100 ng mL–1 of relaxin compared with both the negative and positive control (73.7 ± 2.4, 60.0 ± 4.1, and 60.0 ± 7.1, respectively). A significant (P < 0.05) improvement of cleavage rate was recorded both in the positive control (71.5 ± 4.8) and in the group treated with 100 ng mL–1 of relaxin (70.7 ± 0.5) compared with negative control (52.1 ± 1.5). In conclusion, these preliminary results indicate that relaxin at the concentration of 100 ng mL–1 improves sperm motility, capacitation, and the IVF capability of buffalo sperm.

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IN VITRO EVALUATION OF THE ANTHELMINTIC ACTIVITY OF RHIZOME EXTRACTS OF CURCUMA LONGA (LINN.)

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2018.v11i12.28313 ◽

2018 ◽

Vol 11 (12) ◽

pp. 425

Author(s):

Jyoti Pandey ◽

Suman Mishra ◽

Kamal Jaiswal

Keyword(s):

Curcuma Longa ◽

Anthelmintic Activity ◽

Positive Control ◽

Negative Control ◽

Control Group ◽

Phytochemical Analysis ◽

Alternative Source ◽

Significant Efficacy ◽

Phosphate Buffered Saline

Objective: The current study was carried out to evaluate the anthelmintic activity of the rhizome extract of Curcuma longa as an alternative source of effective remedies for nematodiasis.Methods: The anthelmintic activity of the C. longa was assessed in vitro against Haemonchus spp., a gastrointestinal (abomasum) parasite of goats. Different concentrations of the extracts (1 mg/mL, 2.5 mg/mL, 5 mg/mL, and 10 mg/mL) in phosphate-buffered saline (PBS) were tested, and the results expressed in terms of time of paralysis (minute) and time of death (minute) of the worms. Albendazole (1 mg/mL) was used as a reference (positive control) and PBS as a control group (negative control).Results: The qualitative phytochemical analysis of the methanolic extract (ME) of the plant disclosed the presence of alkaloids, glycosides, terpenoids, flavonoids, tannins, saponins, phenol, anthraquinone, and carbohydrates; whereas, the aqueous extract (AE) showed the presence of alkaloids, carbohydrate, flavonoids, and saponins. Both ME and AE of the C. longa (rhizome) expressed significant efficacy (p≤0.05) in causing paralysis as well as the death of the worms within 12 h of exposure at all tested concentrations, as compared to the negative control. The rhizome extracts of C. longa showed dose-dependent efficacy in causing paralysis of the worm motility and the final progression to death. The results showed that the ME at 10 mg/mL was significantly more potent (p≤0.05) over the AE.Conclusions: This study concluded that the rhizome extract of C. longa exhibited potent anthelmintic efficacy against the nematode parasite, Haemonchus spp.

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Mahkota Dewa (God’s Crown) Fruit Extract Inhibits the Formation of Periodontal Pathogen Biofilms in vitro

Journal of Indonesian Dental Association ◽

10.32793/jida.v2i2.404 ◽

2019 ◽

Vol 2 (2) ◽

pp. 57 ◽

Cited By ~ 1

Author(s):

Diajeng Celia Radita ◽

Armelia Sari Widyarman

Keyword(s):

Brain Heart Infusion ◽

Positive Control ◽

Negative Control ◽

Periodontal Pathogen ◽

Bacterial Strains ◽

Fruit Extract ◽

Fruit Extracts ◽

Level Of Significance ◽

Phosphate Buffered Saline

Introduction: Mahkota dewa (Phaleria macrocarpa) is an Indonesian fruit that contains antibacterial compounds, such as flavonoids, saponins, and tannins; it has been used as an alternative treatment for controlling infection. Objectives: This study aimed to examine the effect of mahkota dewa fruit extract on the formation of Porphyromonas gingivalis (P. gingivalis), Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans), and Treponema denticola (T. denticola) biofilms in vitro. Methods: God’s crown fruit was extracted using the maceration technique, and then diluted into different concentrations (25%, 12.5%, 6.25%, 3.125%, and 1.56%) using phosphate buffered saline (PBS). P. gingivalis ATCC-33277, A. actinomycetemcomitans ATCC-29522, or T. denticola ATCC-35405 were cultured in brain heart infusion (BHI) broth, 24h (anaerobic-condition), and then each type of bacteria (108CFU/mL) was distributed into a 96-well microplate to form a biofilm. Subsequently, the fruit extracts were distributed into the biofilm-containing well plates and incubated for 1h, 6h, and 24h. A biofilm without the fruit extract and chlorhexidine-gluconate (0.2%) was used as the negative and positive control, respectively. Crystal violet (0.5%w/v) was used to determine the density of the remaining biofilm using a microplate spectrophotometer (600 nm). Data were statistically analyzed using one-way ANOVA, and p <0.05 was set as the level of significance. Results: The mahkota dewa fruit extracts significantly inhibited the formation of a biofilm for all three bacterial strains at all concentrations and for each incubation time (p <0.05) based on optical density (OD)±SD. The best concentration of fruit extract to inhibit biofilm formation was 25% for P. gingivalis (OD=0.19±0.06), 12.5% for A. actinomycetemcomitans (OD=0.14 ± 0.16), and 25% for T. denticola (OD=1.17±0.19) in comparison to the biofilm mass of the negative control, which was 1.67±0.06, 1.17±0.34, 2.66±0.38 for P. gingivalis, A. actinomycetemcomitans, and T. denticola, respectively. Conclusion: Based on these results, mahkota dewa fruit extract can inhibit the formation of biofilm on P. gingivalis, A. actinomycetemcomitans, and T. denticola, and it may potentially be used to prevent the infection associated with periodontal disease.

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Evaluation of Mesenchymal Cells (MSC) and Dapsone for the Treatment of Dermonecrotic Wounds Caused by Loxosceles laeta Venom in Rabbits

10.20944/preprints201801.0063.v1 ◽

2018 ◽

Author(s):

Guilherme Martins ◽

Maira Oliveira ◽

Ana Flávia Botelho ◽

Conrado Gamba ◽

Clara Duarte ◽

...

Keyword(s):

Stem Cells ◽

Positive Control ◽

Negative Control ◽

Intradermal Injection ◽

Wound Contraction ◽

Histopathological Analysis ◽

Male Adult ◽

Regenerative Therapies ◽

Phosphate Buffered Saline ◽

Loxosceles Laeta

We studied the efficacy of mesenchymal stem cells (MSC), either alone or associated with dapsone (DAP) in the treatment of dermonecrotic wounds caused by Loxosceles laeta spider venom. Twenty-five male adult rabbits were distributed into five groups, of which four groups received an intradermal injection of 20 μg of L. laeta venom and only one received ultrapure water (negative control). After 4 hours, each group that received venom, was treated with MSC, DAP, MSC + DAP and Phosphate-buffered saline – PBS (positive control). Photographic records were made for analysis of the wound area evolution by morphometry. Twelve days after treatment, the skin samples around the lesion were removed for subsequent histological analysis. Concerning the rate of wound contraction, we observed that DAP showed the best percentage of contraction at day 3. In the treatments using MSCs, a negative value of wound contraction was observed for the isolated MSCs, as well as a lower contraction value for the association of the MSC + DAP when compared to PBS group. Histopathological analysis showed diminished tissue lesion and less intense inflammation in MSCs and DAP groups. This could indicated potential use of stem cells in regenerative therapies after loxoscelic accidents.

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

International Journal of Research in Pharmaceutical Sciences ◽

10.26452/ijrps.v11i3.2385 ◽

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.

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A new normalization for the Nanostring nCounter gene expression assay

10.1101/374173 ◽

2018 ◽

Cited By ~ 1

Author(s):

Ramyar Molania ◽

Johann A. Gagnon-Bartsch ◽

Alexander Dobrovic ◽

Terence P Speed

Keyword(s):

Gene Expression ◽

Single Molecule ◽

Housekeeping Genes ◽

Positive Control ◽

Negative Control ◽

Normalization Method ◽

Data Sets ◽

Analysis Software ◽

Gene Expression Assay ◽

Single Reaction

AbstractThe Nanostring nCounter gene expression assay uses molecular barcodes and single molecule imaging to detect and count hundreds of unique transcripts in a single reaction. These counts need to be normalized to adjust for the amount of sample, variations in assay efficiency, and other factors. Most users adopt the normalization approach described in the nSolver analysis software, which involves background correction based on the observed values of negative control probes, a within-sample normalization using the observed values of positive control probes and normalization across samples using reference (housekeeping) genes. Here we present a new normalization method, Removing Unwanted Variation-III (RUV-III), which makes vital use of technical replicates and suitable control genes. We also propose an approach using pseudo-replicates when technical replicates are not available. The effectiveness of RUV-III is illustrated on four different data sets. We also offer suggestions on the design and analysis of studies involving this technology.

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