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Acute and Repeated 28-Day Oral Dose Toxicity Studies of Thymus vulgaris L. Essential Oil in Rats

  • Rojas-Armas, Juan;Arroyo-Acevedo, Jorge;Ortiz-Sanchez, Manuel;Palomino-Pacheco, Miriam;Castro-Luna, Americo;Ramos-Cevallos, Norma;Justil-Guerrero, Hugo;Hilario-Vargas, Julio;Herrera-Calderon, Oscar
    • Toxicological Research
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    • v.35 no.3
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    • pp.225-232
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    • 2019
  • Thymus vulgaris L. is widely used as an ingredient in cooking and in herbal medicine. However, there is little information about its toxicity. The present study was performed to evaluate the acute and repeated 28-day oral dose toxicity of thyme essential oil in rats. For the acute toxicity test, two groups of three rats were used. The rats received a single dose of essential oil: 300 or 2,000 mg/kg of body weight (bw). The rats were observed individually during the first four hours, and then daily until day 14. For the toxicity test with repeated doses, four groups of 10 rats were used. Doses of 100, 250, and 500 mg/kg/day were tested for 28 days. At the end of the experiment, blood was collected and the animals were sacrificed. Histopathological examination showed that in the lungs of rats given the 2,000 mg/kg bw dose, polymorph nuclear infiltrates, hemosiderin macrophages, and interstitial space thickening were present. In the repeated dose study, all rats survived the 28-day treatment period and apparently showed no signs of toxicity. The hematological and biochemical parameters were not altered. The histopathological study of the organs showed severe changes in the lung, with the dose of 500 mg/kg/day; in the other organs, no alterations were observed or the changes were slight. The body weight was only altered in male rats given the 500 mg/kg dose. The relative weight of the organs did not show any significant changes. Our studies revealed that the essential oil of Thymus vulgaris has moderate oral toxicity according to the results of the acute test, whereas the results of the 28-day oral toxicity test suggest that the no-observed-adverse effect level (NOAEL) is greater than 250 mg/kg/day.

Cytotoxic Effect and Constituent Profile of Alkaloid Fractions from Ethanolic Extract of Ficus septica Burm. f. Leaves on T47D Breast Cancer Cells

  • Nugroho, Agung Endro;Akbar, Fiki Fatihah;Wiyani, Anggie;Sudarsono, Sudarsono
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7337-7342
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    • 2015
  • The study aimed to investigate the profile of alkaloids in two ethyl acetate soluble fractions, namely fractions A and B from an ethanolic extract of Ficus septica leaves and cytotoxic effect on T47D breast cancer cells. Preparation of both fractions involved maceration of leaves with 70% (v/v) ethanol, filtration with $Al_2O_3$, precipitation with 0.1 N HCl, Mayer reagent, and 0.1 N NaOH, and also partition with ethyl acetate. Qualitative thin layer chromatography (TLC) was conducted to determine the profile of alkaloids in the two fractions, using alkaloid specific reagents such as Dragendorff, sodium nitrite, and Van Urk-Salkowski. Cytotoxic effects of both fractions on T47D cells were evaluated using MTT assay with a concentration series of 1.56; 3.12; 6.25; 12.5; 25 and $50{\mu}g/mL$. The TLC test showed that fractions A and B contained alkaloids with Rx values of 0.74 and 0.80 for fraction A and 0.74, 0.84, 0.92 for fraction B with regard to yohimbine using the mobile phase of n-buthanol:glacial acetic acid:distilled water (3:1:1 v/v/v). Moreover, an indole alkaloid was detected with Rx values of 0.80 and 0.84, respectively. Fractions A and B exhibited high cytotoxic effects on T47D cells with IC50 values of 2.57 and $2.73{\mu}g/mL$, respectively. In conclusion, overall the results of this study showed that fractions of Ficus septica contain alkaloids including indole alkaloid or its derivatives and possess a cytotoxic effect on T47D cells. This research supports the idea that alkaloids in F. septica have anticancer activity.

POSSIBILITY OF NONDESTRUCTIVE ANALYSIS OF CHOLESTEROL AND COLLAGEN IN ATHEROSCLEROTIC PLAQUES USING NIRS

  • Neumeister, Volker;Lattke, Peter;Schuh, Dieter;Knuschke, Peter;Reber, Friedemann;Steiner, Gerald;Jaross, Werner
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.4103-4103
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    • 2001
  • The aim of this study was to examine whether near infrared spectroscopy (NIRS) is an acceptable tool to determine cholesterol and collagen in human atherosclerotic plaque without destruction of the analyzed areas and without danger the endothelial cells - three preconditions for the development of a NIR-heart-catheter. The questions were: Can the cholesterol and collagen content of the arterial intima be estimated with acceptable precision in vitro by NIRS despite the matrix inhomogeneity of the plaques and their anatomic variability\ulcorner How deep can such NIR radiation penetrate into arterial tissue without danger for endothelial cells\ulcorner Is this penetration sufficient for information on the lipid and collagen accumulation\ulcorner Using NIRS, cholesterol and collagen can be determined with acceptable precision in model mixtures and human aortic specimens (r=0,896 to 0,957). The chemical reference method was HPLC. The energy dose was 71 mW/$cm^{-2}$ using a fiber optic strand with a length of 1.5m and an optical window of d=4mm. This dose appears to be not dangerous for endothelial cells, It will be attenuated to 50% by a arterial tissue of about 170-$200\mu\textrm{m}$ thickness. The results are also acceptable using a thin coronary catheter-like fiber optic strand (d=1mm).

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Effect of Root Extracts of Medicinal Herb Glycyrrhiza glabra on HSP90 Gene Expression and Apoptosis in the HT-29 Colon Cancer Cell Line

  • Nourazarian, Seyed Manuchehr;Nourazarian, Alireza;Majidinia, Maryam;Roshaniasl, Elmira
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8563-8566
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    • 2016
  • Colorectal cancer is one of the most common lethal cancer types worldwide. In recent years, widespread and large-scale studies have been done on medicinal plants for anti-cancer effects, including Glycyrrhiza glabra. The aim of this study was to evaluate the effects of an ethanol extract Glycyrrhiza glabra on the expression of HSP90, growth and apoptosis in the HT-29 colon cancer cell line. HT-29 cells were treated with different concentrations of extract (50,100,150, and $200{\mu}g/ml$). For evaluation of cell proliferation and apoptosis, we used MTT assay and flow cytometry technique, respectively. RT-PCR was also carried out to evaluate the expression levels of HSP90 genes. Results showed that Glycyrrhiza glabra inhibited proliferation of the HT-29 cell line at a concentration of $200{\mu}g/ml$ and this was confirmed by the highest rate of cell death as measured by trypan blue and MTT assays. RT-PCR results showed down-regulation of HSP90 gene expression which implied an ability of Glycyrrhiza glabra to induce apoptosis in HT-29 cells and confirmed its anticancer property. Further studies are required to evaluate effects of the extract on other genes and also it is necessary to make an extensive in vivo biological evaluation and subsequently proceed with clinical evaluations.

Polymorphisms in TP53 (rs1042522), p16 (rs11515 and rs3088440) and NQO1 (rs1800566) Genes in Thai Cervical Cancer Patients with HPV 16 Infection

  • Chansaenroj, Jira;Theamboonlers, Apiradee;Junyangdikul, Pairoj;Swangvaree, Sukumarn;Karalak, Anant;Chinchai, Teeraporn;Poovorawan, Yong
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.341-346
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    • 2013
  • The risk of cervical cancer development in women infected with HPV varies in relation to the individual host's genetic makeup. Many studies on polymorphisms as genetic factors have been aimed at analyzing associations with cervical cancer. In this study, single nucleotide polymorphisms (SNPs) in 3 genes were investigated in relation to cervical cancer progression in HPV16 infected women with lesions. Two thousand cervical specimens were typed by PCR sequencing methods for TP53 (rs1042522), p16 (rs11515 and rs3088440) and NQO1 (rs1800566). Ninety two HPV16 positive cases and thirty two normal cases were randomly selected. Analysis of TP53 (rs1042522) showed a significantly higher frequency in cancer samples (OR=1.22, 95%CI=1.004-1.481, p-value=0.016) while differences in frequency were not significant within each group (p-value=0.070). The genotype distributions of p16 (rs11515 and rs3088440) and NQO1 (rs1800566) did not show any significantly higher frequency in cancer samples (p-value=0.106, 0.675 and 0.132, respectively) or within each group (p-value=0.347, 0.939 and 0.111, respectively). The results indicated that the polymorphism in TP53 (rs1042522) might be associated with risk of cervical cancer development in HPV16 infected women. Further studies of possible mechanisms of influence on cervical cancer development would be useful to manage HPV infected patients.

A new biomarker for the early diagnosis of ovarian torsion: SCUBE-1

  • Uyanikoglu, Hacer;Hilali, Nese Gul;Yardimciel, Mesut;Koyuncu, Ismail
    • Clinical and Experimental Reproductive Medicine
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    • v.45 no.2
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    • pp.94-99
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    • 2018
  • Objective: Prompt diagnosis and management are essential for saving the adnexal organs from infarction in cases of ovarian torsion (OT). This study aimed to determine the diagnostic significance of signal peptide, complement C1r/C1s, Uegf, and Bmp1 (CUB), and epidermal growth factor-like domain-containing protein-1 (SCUBE-1) levels in cases of OT, an emergent ischemic condition, and the relationship of SCUBE-1 with oxidative stress parameters. Methods: This prospective study was conducted among 15 OT patients and 20 age- and gravidity-matched healthy women. SCUBE-1 serum concentrations were determined by using enzyme-linked immunosorbent assays. In addition, oxidative stress was evaluated by measuring the serum levels of advanced oxidation protein products (AOPP), ferric reducing ability of plasma (FRAP), and glutathione (GSH). Results: The SCUBE-1 titers were significantly higher in the patients with OT than in the controls (p=0.008). In addition, serum FRAP and GSH levels were significantly lower in the OT patients than in the controls (p<0.001 for both). Serum AOPP levels were higher in the OT patients, but this trend was not statistically significant (p>0.05). Furthermore, there were no correlations between SCUBE-1 levels and age, gravidity, parity, cyst size, and AOPP, FRAP, or GSH levels (p>0.05). Conclusion: We believe that SCUBE-1 may be a promising biomarker for the early diagnosis of OT.

Single Nucleotide Polymorphism of Interferon Lambda-4 Gene is not Associated with Treatment Response to Pegylated Interferon in Thai Patients with Chronic Hepatitis B

  • Limothai, Umaporn;Wasitthankasem, Rujipat;Poovorawan, Yong;Tangkijvanich, Pisit
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.13
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    • pp.5515-5519
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    • 2015
  • The single nucleotide polymorphism (SNP) ss469415590 in the interferon lambda-4 (IFNL4) gene has recently been reported to have an association with treatment response in chronic hepatitis C. However, any importance of the SNP in association with response to pegylated interferon (PEG-IFN) therapy in patients with chronic hepatitis B (CHB) is unclear. We retrospectively analyzed data for Thai patients with CHB treated with PEG-IFN for 48 weeks. Virological response (VR) for HBeAg-positive CHB was defined as HBeAg seroconversion plus HBV DNA level <2,000 IU/mL at 24 weeks post-treatment. VR for HBeAg-negative CHB was defined as an HBV DNA level <2,000 IU/mL at 48 weeks. The SNP was identified by real time PCR using the TaqMan genotyping assay with MGB probes. A total 254 patients (107 HBeAg-positive and 147 HBeAg-negative) were enrolled in the study. The distribution of TT/TT, ${\Delta}G/TT$ and ${\Delta}G/{\Delta}G$ genotypes was 221 (87.0%), 32 (12.6%) and 1 (0.4%), respectively. Patients with non-TT/TT genotypes had significantly higher baseline HBV DNA levels than patients with the TT/TT genotype. In HBeAg-positive CHB, 41.2% of patients with TT/TT genotype versus 50.0% with non-TT/TT genotype achieved VR (P=0.593). In HBeAg-negative CHB, the corresponding figures were 40.3% and 43.5%, respectively (P=0.777). There was no significant correlation between the SNP genotypes and HBsAg clearance in both groups of patients. In summary, ss469415590 genotypes were not associated with response to PEG-IFN in Thai patients with HBeAg-positive and HBeAg-negative CHB.

Osseous changes in the temporomandibular joint in rheumatoid arthritis: A cone-beam computed tomography study

  • Rehan, Ola Mohamed;Saleh, Hoda Abdel Kader;Raffat, Hala Ahmed;Abu-Taleb, Noha Saleh
    • Imaging Science in Dentistry
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    • v.48 no.1
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    • pp.1-9
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    • 2018
  • Purpose: To evaluate osseous changes of temporomandibular joint (TMJ) in patients with rheumatoid arthritis (RA) using cone-beam computed tomography (CBCT) and to correlate the imaging findings with the severity of TMJ dysfunction, clinical findings, and laboratory findings. Materials and Methods: This study consisted of 28 subjects, including 14 RA patients and 14 controls, who were scheduled to undergo CBCT imaging for the diagnosis of a complaint not related to or affecting the TMJ. The Fonseca's questionnaire was used to assess the severity of TMJ dysfunction. Rheumatoid factor (RF) and the erythrocyte sedimentation rate (ESR) were assessed in the RA patients. CBCT was then performed in all subjects and osseous TMJ abnormalities were assessed. Results: According to the Fonseca's questionnaire, 14.3% of the patients had no TMJ dysfunction, while 50%, 21.4%, and 14.3% had mild, moderate, and severe dysfunction, respectively. RF was positive in 64.3% of patients, and the ESR level was high in 100%. Imaging findings revealed a statistically significantly higher prevalence of erosion (85.7%), flattening (89.3%), osteophyte formation (32.1%), subchondral cyst (32.1%), sclerosis (64.3%), and condylar irregularities (28.6%) in the RA patients than in the controls. No correlations were found between CBCT findings and the clinical findings, the severity of TMJ dysfunction, disease duration, or laboratory results. Conclusion: RA patients might show extensive osseous abnormalities with no/mild clinical signs or symptoms of TMJ dysfunction that necessitate TMJ imaging for these patients. CBCT is a valuable and efficient modality that can assess osseous TMJ changes in RA patients.

A comparative study of the prevalence of Helicobacter pylori in the oral biofilms of a group of dental and non-dental undergraduates from Sri Lanka

  • Mallikaarachchi, MADKS;Rajapakse, Sanath;Gunawardhana, KSND;Jayatilake, JAMS
    • International Journal of Oral Biology
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    • v.46 no.1
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    • pp.60-65
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    • 2021
  • Dental health care workers (DHCW) are at a risk of occupational exposure to Helicobacter pylori from the aerosolized oral biofilms and saliva of patients. We designed this study to investigate the prevalence of H. pylori in the oral biofilms of a group of dental and non-dental undergraduates from Sri Lanka. After obtaining informed consent, oral biofilms were collected from 38 dental undergraduates (19 males and 19 females) undergoing clinical training and 33 non-dental undergraduates (14 males and 19 females). The participants were in the age range of 22-27 years and had healthy periodontium. Total DNA from the oral biofilms were extracted, and H. pylori DNA was detected using polymerase chain reaction (PCR) amplification of 16S rRNA gene of H. pylori using JW22-JW23 primers, and the results were confirmed using PCR amplification of H. pylori-urease specific HPU1-HPU2 primers. Out of 71 participants, 11 (28.95%) dental and 3 (9.09%) non-dental undergraduates had H. pylori in their oral biofilms indicating an overall prevalence rate of 19.72% (14/71). Thus, the prevalence of H. pylori in oral biofilms was significantly higher in dental undergraduates than in non-dental undergraduates (p < 0.05). An odds ratio of 4.07 indicated that dental undergraduates were four times more likely to harbor H. pylori in their oral biofilms than non-dental undergraduates. Foregoing data support the fact that there may be greater occupational risk of exposure to H. pylori for dental undergraduates during clinical training than that for non-dental undergraduates, warranting meticulous infection control practices during clinical dentistry.

Surgical outcomes in dogs with tracheal collapse treated with a novel cross-and-hook braided endoluminal stent

  • Uemura, Akiko;Ozai, Yusuke;Hamabe, Lina;Yoshida, Tomohiko;Tanaka, Ryou
    • Journal of Veterinary Science
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    • v.23 no.3
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    • pp.46.1-46.8
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    • 2022
  • Background: Stenting is an effective treatment option for tracheal collapse in dogs. Cross-braided tracheal stents are currently the norm in veterinary medicine, but cross-and-hook braided stents have recently been adopted in human medicine. We examined whether stents manufactured using this novel braiding technique provided additional advantages for the treatment of tracheal collapse in dogs. Objectives: To evaluate the outcomes of cross-and-hook braided stent implantation in the treatment of tracheal collapse in dogs. Methods: The medical records of 22 client-owned dogs that underwent luminal placement of cross-and-hook braided Fauna Stents for the treatment of tracheal collapse between January 2018 and July 2021 were examined and data on canine signalment, clinical signs, diagnostic test results, surgical outcomes, and postoperative complications were retrieved and analyzed statistically. Results: Twenty-six stents were surgically implanted, with 20 dogs (90.9%) receiving one stent and the remaining two (9.1%) receiving two or more stents. All dogs survived the procedure. The median survival time at a median follow-up of 990 days was 879 days. At the final follow-up examination, loss or mild improvement of cough was observed in all dogs. Conclusions: Compared with conventional lumen stents, the cross-and-hook braided Fauna Stent offered a higher survival rate and improved clinical symptoms in all patients. The results of this study suggest that the Fauna Stent may be a promising treatment option for dogs with tracheal collapse.