scholarly journals Preliminary Assessment of DNA Profile Patterns of Neuronal Differentiation Factor (NeuroD1) Gene in the Hippocampus of Wistar Rats (Rattus norvegicus) Ingested Hyoscyamine Fraction of Datura stramonium Seeds

2020 ◽  
pp. 1-6
Author(s):  
Idris Abdu Tela ◽  
◽  
Sunday Abraham Musa ◽  
Ibrahim Abdullahi Iliya ◽  
◽  
...  

The study was designed to find out how oral ingestion of hyoscyamine fraction of D. stramonium seeds affect the DNA profiles patterns and expression of NeuroD1 gene in the hippocampus of adult Wistar rats. Fresh D. stramonium seeds were procured, identified, macerated and fractionated. Thirty-two (32) male adult Wistar rats weighed 200-250 grams were procured and divided into four groups comprised of control and three treatment groups for the study. The control received an equivalent bodyweight of normal saline, while the treatment groups received 200, 400 and 800 mg/kgbwt of hyoscyamine fraction for three weeks respectively. The animals were sacrificed, tissues harvested for histology, gel electrophoresis and gene expression studies. The fold changes were calculated and DNA profiles were determined using LabImage bio-imaging software. One-way ANOVA followed by Fisher’s multiple comparisons post-hoc test was used to obtain the difference in the expression of the gene across the groups using Minitab 17 (LLC., U.K.) statistical package software. P < 0.05 was considered statistically significant. There was a statistically significant downregulation (p = 0.001) in the NeuroD1 expression, increased DNA fragment size, decreased DNA migration distance and band volume, and necrosis of the dentate gyrus (DG) region in the hippocampus of the treated animals. In conclusion, oral ingestion of hyoscyamine fraction of D. stramonium seeds is toxic to the DNA profile patterns and expression of the NeuroD1 gene in Wistar rats.

2018 ◽  
Vol 1 (2) ◽  
pp. 1-17
Author(s):  
Fahcreza ◽  
Elsa Iskandar ◽  
Rachmat Hidayat ◽  
Petty Purwanita ◽  
Anang Tribowo ◽  
...  

Abstract Background: Chemical trauma to the cornea is an emergency condition of the eye that requires early diagnosis and good treatment. Alkaline have ability to saponify fatty acids in cells and cell membranes which can make penetration into the stroma and destroy proteoglycans and collagen in cells. Aloe vera (AV) contains several active substances that are reported to have anti-inflammatory, immunomodulatory, and wound healing effects. AV has been reported to accelerate the healing process of corneal epithelial defects by increasing fibroblast proliferation, collagen production and growth factor production. This study aims to determine the difference between the effect of aloe vera extract with a concentration of 10%, 20%, 40% and BSS on the healing of extensive corneal lesions in white wistar rats alkaline trauma models. Method: This study was an experimental study with a pre and posttest only with control group design in vivo approach to 30 Wistar white rats which were divided into 5 treatment groups for 3 days. Comparative analysis of effectiveness using the ANNOVA test or the Kruskal Wallis test and continued by the post hoc test. Results: Based on the one way ANOVA test there was a statistically significant difference in effectiveness between the five treatment groups on the percentage of corneal wound healing area and TGF-β expression with an assessment of p = 0,000 each. The administration of alloevera (AV) concentration of 20% had a significant difference in percentage of healing of corneal lesions and TGF-β expression compared with other treatment groups with p = 0,000 each. Large differences in the area of corneal lesions in the 40% AV group were -0.45 in the BBS group, 0.146 in the 10% AV group, 0.493 in the 20% AV group. The difference in the AV group 10% was 0.30 in the BBS group, -064 in the AV group 20%, and -0.14 in the AV group 40%. However, TGFβ expression in the normal control group that did not receive treatment was 54.94 (53.21-56-12). TGFβ levels in the BSS group were 10.44, the 10% aloe vera group was 25.43, 47.99 for the 20% aloe vera group and 37.95 for the 40% aloe vera group. Conclusion: There is a difference between the effect of aloe vera extract with concentrations of 10%, 20%, 40% and BSS on the extensive healing of corneal lesions in white wistar rats with alkaline chemical trauma models.


2019 ◽  
Vol 26 (2) ◽  
Author(s):  
Puruhito Eko Nugroho ◽  
Fikri Rizaldi ◽  
Wahjoe Djati Soesanto

Objective: Smoking are known to have detrimental effect on sperm. The effects are suspiciously come from nicotine as its main pharmacologic component. This study is carried out to analyze the effect of nicotine exposure per inhalation to the difference of motile sperm percentage and normal morphology sperm percentage of male wistar rats. Material & Methods: Thirty male rats were used for this research. They were randomly divided into 5 groups and treated with nicotine inhalation 0.5 mg/kgBW,  1 mg/kgBW, 2 mg/kgBW and 4 mg/kgBW for 30 days while the control rats were not given any treatment. At the end of experiment, sperm analysis was carried out to measure the percentage of motile sperm and percentage of sperm with normal morphology. Results: Percentage of motile sperm significantly decreased (p<0.05) in all treatment groups compared to control group. Comparison between treatment groups revealed that the percentage of motile sperm significantly decreased in 0.5 mg/kgBW vs 2 mg/kgBW, 0.5 mg/kgBW vs 4 mg/kgBW, and 1 mg/kgBW vs 4 mg/kgBW. Percentage of normal morphology sperm decreased significantly (p<0.05) in 1 mg/kgBW, 2 mg/kgBW, and 4 mg/kgBW compared to control groups. Comparison between treatment groups revealed that there were no significant decreased between each treatment groups. Conclusion: This research conclude that nicotine exposure per inhalation can deteriorate the percentage of motile sperm and the percentage of normal morphology sperm of male wistar rats.


2013 ◽  
Vol 118 (5) ◽  
pp. 1160-1169 ◽  
Author(s):  
Delia Aguado ◽  
Mariana Abreu ◽  
Javier Benito ◽  
Javier Garcia-Fernandez ◽  
Ignacio A. Gómez de Segura

Abstract Background: Opioid antagonists at ultra-low doses have been used with opioid agonists to prevent or limit opioid tolerance. The aim of this study was to evaluate whether an ultra-low dose of naloxone combined with remifentanil could block opioid-induced hyperalgesia and tolerance under sevoflurane anesthesia in rats. Methods: Male adult Wistar rats were allocated into one of four treatment groups (n = 7), receiving remifentanil (4 µg·kg−1·min−1) combined with naloxone (0.17 ng·kg−1·min−1), remifentanil alone, naloxone alone, or saline. Animals were evaluated for mechanical nociceptive thresholds (von Frey) and subsequently anesthetized with sevoflurane to determine the baseline minimum alveolar concentration (MAC). Next, treatments were administered, and the MAC was redetermined twice during the infusion. The experiment was performed three times on nonconsecutive days (0, 2, and 4). Hyperalgesia was considered to be a decrease in mechanical thresholds, whereas opioid tolerance was considered to be a decrease in sevoflurane MAC reduction by remifentanil. Results: Remifentanil produced a significant decrease in mechanical thresholds compared with baseline values at days 2 and 4 (mean ± SD, 30.7 ± 5.5, 22.1 ± 6.4, and 20.7 ± 3.7g at days 0, 2, and 4, respectively) and an increase in MAC baseline values (2.5 ± 0.3, 3.0 ± 0.3, and 3.1 ± 0.3 vol% at days 0, 2, and 4, respectively). Both effects were blocked by naloxone coadministration. However, both remifentanil-treated groups (with or without naloxone) developed opioid tolerance determined by their decrease in MAC reduction. Conclusions: An ultra-low dose of naloxone blocked remifentanil-induced hyperalgesia but did not change opioid tolerance under inhalant anesthesia. Moreover, the MAC increase associated with hyperalgesia was also blocked by naloxone.


Author(s):  
В.Н. Сахаров ◽  
П.Ф. Литвицкий ◽  
Е.И. Алексеева ◽  
Н.А. Маянский ◽  
Р.Ш. Закиров

Цель исследования - изучение перепрограммирования мононуклеарных лейкоцитов на модели системного ювенильного идиопатического артрита (сЮИА), воспроизводимой у крыс Wistar с использованием полного адъюванта Фрейнда и липополисахарида. Методика. сЮИА воспроизведен у 6-месячных крыс-самцов Wistar. На 40-е сут. эксперимента животные были разделены на 3 группы: 1-я группа - контроль; 2-я - группа доксициклина; 3-я - группа дексаметазона. Взятие проб крови у животных проводили на нулевые, 41-е и 55-е сут. Мононуклеарные клетки периферической крови выделяли гравиметрически, после чего окрашивали их на маркеры и внутриклеточные цитокины. Дифференцировали моноциты (CD3-CD4+) и Т-хелперы (CD3+CD4+). Анализировали динамику внутриклеточной экспрессии интерлейкина IL-4 (рассматривали как маркер про-М2 фенотипа, так как в случае выделения из клетки ИЛ-4 служит стимулятором М2 поляризации макрофагов) и IFN-g (как маркер про-М1 фенотипа) по данным проточной цитофлуориметрии. Применяли непараметрический статистический тест Mann-Whitney-Wilcoxon в программе R для статистической обработки данных. Результаты и заключение. При моделировании сЮИА выявлено закономерное изменение фенотипа моноцитов. Применение же доксициклина и дексаметазона приводило к более ранней поляризации их по про-М2-пути в отношении моноцитов (на 41-е сут.) в сравнении с контролем. Про-М1 эффект (на 55-е сут., в сравнении с контролем) выявлен также в группах доксициклина и дексаметазона. У животных разных групп обнаружены характерные динамические изменения внутриклеточной экспрессии цитокинов. Важно, что различная направленность поляризации фенотипа при сЮИА и применении препаратов наблюдается не только у моноцитов, но и у Т-хелперов. The study objective was to evaluate targeted reprogramming of peripheral blood mononuclear cells in systemic juvenile idiopathic arthritis (sJIA) modeled in 6-month-old male Wistar rats by co-administration of complete Freund’s adjuvant and lipopolysaccharide. Methods. On day 40 of the experiment, rats were divided into three groups: control, doxycycline, and dexamethasone groups. Blood samples were collected on days 0, 41, and 55. Peripheral blood mononuclear cells were isolated gravimetrically and stained for markers and cytokines. Monocytes (CD3-CD4+) and T-helpers (CD3+CD4+) were differentiated as target cells. IL-4 was considered a marker for the pro-M2 phenotype since IL-4 can activate M2 macrophage polarization; IFN-g was considered a marker for the pro-M1 phenotype. Time-related changes in the intracellular expression of IL-4 and IFN-g were studied using flow cytometry. Data were analyzed using nonparametric statistical tests (Mann-Whitney-Wilcoxon) in the R environment for statistical computing. Results and conclusions. Monocytes (like macrophages) underwent reprogramming during the development of modeled sJIA disease. In monocytes of doxycycline and dexamethasone treatment groups, pro-M2 effects were observed earlier (day 41) than in the control group. Pro-M1 effects were observed in monocytes of doxycycline and dexamethasone groups on day 55, as compared with the control group. Characteristic time-related changes of intracellular cytokine expression were described for different groups. Importantly, the differently directed phenotype polarization was observed in sJIA and treatment groups for both monocytes and T-helpers.


2020 ◽  
Vol 16 ◽  
Author(s):  
Seiji Umemoto ◽  
Toshio Ogihara ◽  
Masunori Matsuzaki ◽  
Hiromi Rakugi ◽  
Kazuyuki Shimada ◽  
...  

Background: In the trial known as COPE (Combination Therapy of Hypertension to Prevent Cardiovascular Events) three benidipine (a calcium channel blocker; CCB) regimens were compared. Hypertensive Japanese outpatients aged 40–85 years (n=3,293) who did not achieve the target blood pressure of <140/90 mmHg with benidipine 4 mg/day were treated with the diuretic thiazide (n=1,094) or a β-blocker (n=1,089) or an additional angiotensin receptor blocker (ARB; n=1,110). A significantly higher incidence of hard cardiovascular composite endpoints and of fatal or non-fatal strokes was observed in the benidipine-β-blocker group compared to the benidipine-thiazide group. Objective and Methods: We further evaluated the treatment effects of the three benidipine-based regimens on vascular and renal events in a sub-analysis of the COPE patients. Results: A total of 10 vascular events (0.8 per 1,000 person-years) including one aortic dissection (0.1 per 1,000 person-years) and nine cases of peripheral artery disease (0.8 per 1,000 person-years) were documented, as was a total of seven renal events (0.6 per 1,000 person-years). No significant differences in vascular and renal events were revealed among the three treatment groups: vascular events p=0.92 renal events p=0.16 log-rank test. Conclusions: Blood pressure-lowering therapy with benidipine combined with an ARB, β-blocker, or thiazide was similarly effective in the prevention of vascular and renal events in hypertensive outpatients, although there is no enough these events to compare the difference in the three treatment groups.


2021 ◽  
Vol 11 (4) ◽  
pp. 1943
Author(s):  
Joo-Young Kim ◽  
Ju Yeon Jung ◽  
Da-Hye Kim ◽  
Seohyun Moon ◽  
Won-Hae Lee ◽  
...  

Analytical techniques such as DNA profiling are widely used in various fields, including forensic science, and novel technologies such as direct polymerase chain reaction (PCR) amplification are continuously being developed in order to acquire DNA profiles efficiently. However, non-specific amplification may occur depending on the quality of the crime scene evidence and amplification methods employed. In particular, the ski-slope effect observed in direct PCR amplification has led to inaccurate interpretations of the DNA profile results. In this study, we aimed to reduce the ski-slope effect by using dimethyl sulfoxide (DMSO) in direct PCR. We confirmed that DMSO (3.75%, v/v) increased the amplification yield of large-sized DNA sequences more than that of small-sized ones. Using 50 Korean buccal samples, we further demonstrated that DMSO reduced the ski-slope effect in direct PCR. These results suggest that the experimental method developed in this study is suitable for direct PCR and may help to successfully obtain DNA profiles from various types of evidence at crime scenes.


Cells ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1336
Author(s):  
Caterina Catalano ◽  
Loredana Abbate ◽  
Antonio Motisi ◽  
Dalila Crucitti ◽  
Vincenzo Cangelosi ◽  
...  

Polyploidy plays an important role in plant adaptation to biotic and abiotic stresses. Alterations of the ploidy in grapevine plants regenerated via somatic embryogenesis (SE) may provide a source of genetic variability useful for the improvement of agronomic characteristics of crops. In the grapevine, the SE induction process may cause ploidy changes without alterations in DNA profile. In the present research, tetraploid plants were observed for 9.3% of ‘Frappato’ grapevine somatic embryos regenerated in medium supplemented with the growth regulators β-naphthoxyacetic acid (10 µM) and N6-benzylaminopurine (4.4 µM). Autotetraploid plants regenerated via SE without detectable changes in the DNA profiles were transferred in field conditions to analyze the effect of polyploidization. Different ploidy levels induced several anatomical and morphological changes of the shoots and mature leaves. Alterations have been also observed in stomata. The length and width of stomata of tetraploid leaves were 39.9 and 18.6% higher than diploids, respectively. The chloroplast number per guard cell pair was higher (5.2%) in tetraploid leaves. On the contrary, the stomatal index was markedly decreased (12%) in tetraploid leaves. The observed morphological alterations might be useful traits for breeding of grapevine varieties in a changing environment.


Author(s):  
Eden D Alamaw ◽  
Benjamin D Franco ◽  
Katechan Jampachaisri ◽  
Monika K Huss ◽  
Cholawat Pacharinsak

A new extended-release buprenorphine (XR), an FDA-indexed analgesic, has recently become available to the laboratoryanimal community. However, the effectiveness and dosing of XR has not been extensively evaluated for rats. We investigatedXR’s effectiveness in attenuating postoperative hypersensitivity in a rat incisional pain model. We hypothesized that highdose of XR would attenuate mechanical and thermal hypersensitivity more effectively than the low dose of XR in this model. We performed 2 experiments. In experiment 1, male adult Sprague–Dawley rats (n = 31) were randomly assigned to 1 of the 4 treatment groups: 1) saline (saline, 0.9% NaCl, 5 mL/kg, SC, once); 2) sustained-release buprenorphine (Bup-SR; 1.2 mg/kg, SC, once), 3) low-dose extended-release buprenorphine (XR-Lo; 0.65 mg/kg, SC, once), and 4) high-dose extended-releasebuprenorphine (XR-Hi; 1.3 mg/kg, SC, once). After drug administration, a 1 cm skin incision was made on the plantar hind paw under anesthesia. Mechanical and thermal hypersensitivity were evaluated 1 d before surgery (D-1), 4 h after surgery (D0), and for 3 d after surgery (D1, D2, and D3). In experiment 2, plasma buprenorphine concentration (n = 39) was measured at D0, D1, D2, and D3. Clinical observations were recorded daily, and a gross necropsy was performed on D3. Mechanical and thermal hypersensitivity were measured for 3 d (D0-D3) in the saline group. Bup-SR, XR-Lo, and XR-Hi effectively attenuated mechanical hypersensitivity for D0-D3. Plasma buprenorphine concentrations remained above 1 ng/mL on D0 and D1 in all treatment groups. No abnormal clinical signs were noted, but injection site reactions were evident in the Bup-SR (71%), XR-Lo (75%), and XR-Hi (87%) groups. This study indicates that XR-Hi did not attenuate hypersensitivity more effectivelythan did XR-Lo in this model. XR 0.65 mg/kg is recommended to attenuate postoperative mechanical hypersensitivity for upto 72 h in rats in an incisional pain model.


Author(s):  
Rizka Veni ◽  
Awal Prasetyo ◽  
Muflihatul Muniroh

This study aims to analyze the effect of combination of motor vehicle particular matter exposure and high-fat diet in kidney histopathology, creatinine levels, and MDA levels in Wistar rats. This study used a posttest-only control group design. Eighteen healthy male Wistar rats were divided into three groups. The intervention groups received motor vehicle fume exposure for 100 s with normal diet (X1) or high-fat diet (X2), and the control group received no exposure (C). Data analysis was processed with a SPSS 25.0 computer program by using the one-way ANOVA test followed by post hoc LSD. The degree of kidney histopathological damage showed significant differences between the X1 and X2 groups when compared with the control group (p < 0.05). The results of the creatinine level examination found a significant difference between the X2 and C groups (p < 0.05) and the treatment groups X1 and X2 (p < 0.05). The results of kidney MDA level examination showed a significant difference between the treatment groups (X1 and X2) and the control group (p < 0.05). The combination of particular matter of motor vehicle fumes exposure and high-fat diet could induce kidney damage through histopathological change and increased creatinine levels and kidney MDA levels in Wistar rats.


2018 ◽  
Vol 23 (5) ◽  
pp. 277-284 ◽  
Author(s):  
Chia-Huei Chu ◽  
Shiau-Ru Chiou ◽  
Mao-Che Wang ◽  
An-Suey Shiao ◽  
Tzong-Yang Tu ◽  
...  

Objective: The purpose of this retrospective study was to investigate the difference in treatment outcomes for patients with idiopathic sudden sensorineural hearing loss (SSNHL) undergoing concurrent or sequential intravenous (IV) and intratympanic (IT) steroid therapies. Methods: Patients with idiopathic SSNHL admitted to Taipei Veterans Hospital from August 2011 to August 2012 were enrolled. Patients were treated with both IV dexamethasone 5 mg b.i.d. for 5 days, then tapered over 6 days, and IT injections of dexamethasone 5 mg daily. The administration of IV and IT steroids was given either concurrently or sequentially (IV steroid was administered from days 1–5 followed by IT steroid treatment starting on day 4 or day 5). The hearing outcomes of the concurrent and sequential groups were analyzed. Results: Overall, after ≥2 months following treatment, across frequencies ranging from 250 to 8,000 Hz and pure-tone average (PTA) assessments, hearing improvements were similar between treatment groups, except at the frequencies of 4,000 and 8,000 Hz where the concurrent treatment group had greater hearing gain than the sequential group (4,000 Hz: 30.68 ± 28.96 vs. 14.52 ± 24.06 dB, respectively, p = 0.042; 8,000 Hz: 22.62 ± 23.59 vs. 7.67 ± 21 dB, p = 0.030). Across frequencies and PTA assessments, a similar percentage of patients had ≥20-dB gains in hearing compared with patients treated sequentially, except at 8,000 Hz where a greater percentage of patients in the concurrent group (57.1%) than the sequential group (23.3%) (p = 0.014) had ≥20-dB hearing gains. Conclusion: The findings suggest that both concurrent and sequential treatment improve hearing in patients with idiopathic SSNHL, and that concurrent treatment may show greater benefit than sequential therapy, particularly at high frequencies.


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