Temporal and Spatial Expression Characteristics of MiR-155 and Rheb/mTOR Signaling Pathway in Ischemia–Reperfusion Injury of Rats

Abstract Backgrouds: Stroke is the second most prevalent cause of death and the first cause of longterm disability worldwide. Inhibition of miR-155 was found playing a protective role in ischemic stroke, one possible mechanism was regulating Ras-homolog enriched in brain (Rheb)/mammalian target of rapamycin (mTOR) pathway. For possible specific intervention strategy, further exploring the expression characteristics of miR-155 and mRNAs of the Rheb/mTOR pathway in ischemic stroke is neccesary. Results: Our results demonstrated that the infarction volume decreased with the prolongation of the reperfusion in the MCAO/R model rats (P < 0.05). Meanwhile, the miR155 expression obviously increased in both the ischemic core and the ischemic penumbra (IP) area of the model rats, but this trend weakened as the reperfusion time increased. Besides, the expression of mRNAs of Rheb, mTOR, S6kb1, and 4Ebp1 seemed to increase in both the ischemic core and the IP area of the model rats.Interestingly, the mRNA level of S6kb1 obviously increased of all model groups in both the ischemic core and the IP area (P < 0.05),while the mRNA levels of Rheb, mTOR, and 4Ebp1 increased in the first 24 h and rapidly decreased after 48 h and as a result, a statistically significant difference was found only in the 48-h group (P < 0.05). Conclusion: Along with the shrinked infarct volume, the levels of miR-155 decreased and the S6kb1 mRNA level increased as the leghtening of re-perfusion, as to the mRNA levels of Rheb, mTOR, and 4Ebp1,statistical significance was found only in the 48-h group. Unexpectedly, there was no difference between the ischemic core and the IP area for all the above molecules.Indicating that intervention measures targeting to miR155 should be taken systemicly as early as possible after stroke onset,especially within the early 48 hours.

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Increased NOX1 and DOUX2 Expression in the Colonic Mucosa of Patients with Chronic Functional Constipation

10.21203/rs.3.rs-643716/v1 ◽

2021 ◽

Author(s):

Xiuqin Wei ◽

Chunbo Kang ◽

Lei Gao ◽

Mengqiao Zhang ◽

Mei Xue ◽

...

Keyword(s):

Nadph Oxidase ◽

Healthy Subjects ◽

Mrna Level ◽

Functional Constipation ◽

Inflammatory Cells ◽

Histological Structure ◽

Mrna Levels ◽

Significant Difference ◽

And Control ◽

Colonic Biopsies

Abstract Aim To determine whether oxidative stress and inflammation are associated with constipation by examining the expression of the main producers of reactive oxygen species, NADPH oxidases, and pro-inflammatory cytokines in the colon of patients with chronic functional constipation. Methods The colonic biopsies were collected from 32 patients with chronic functional constipation and 30 healthy subjects who underwent colonoscopy. Colonic mucosal histology was observed. IL-1β, IL-6, IL-8 mRNA, and four members of NADPH oxidase (NOX1, NOX2, DOUX2 and NOX4) protein and mRNA were assessed by immunohistochemistry, western blotting and RT-PCR. Results The tissues from both patients and healthy subjects showed normal histological structure without increase of inflammatory cells. NOX1 protein and mRNA levels were significantly increased compared to controls (P<0.05). DOUX2 protein, but not mRNA, was increased by twofold compared to controls (P<0.05). The levels of NOX2 and NOX4 protein and mRNA demonstrated no significant difference between patients and control subjects. The levels of IL-1β and IL-6 mRNA were significantly higher in constipation patients (P<0.05), while IL-8 mRNA level was no different between the two groups. Conclusion NADPH oxidase and pro-inflammatory cytokine might be involved in the pathogeneses of chronic functional constipation.

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Differential response to myocardial reperfusion injury in eNOS-deficient mice

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00855.2001 ◽

2002 ◽

Vol 282 (6) ◽

pp. H2422-H2426 ◽

Cited By ~ 55

Author(s):

Brent R. Sharp ◽

Steven P. Jones ◽

David M. Rimmer ◽

David J. Lefer

Keyword(s):

Ischemia Reperfusion ◽

Myocardial Necrosis ◽

In Vivo Model ◽

Pcr Analysis ◽

Inos Mrna ◽

Mrna Levels ◽

Wild Type ◽

Significant Difference ◽

Deficient Mice

Two strains of endothelial nitric oxide synthase (eNOS)-deficient (−/−) mice have been developed that respond differently to myocardial ischemia-reperfusion (MI/R). We evaluated both strains of eNOS−/− mice in an in vivo model of MI/R. Harvard (Har) eNOS−/− mice ( n = 12) experienced an 84% increase in myocardial necrosis compared with wild-type controls ( P < 0.05). University of North Carolina (UNC) eNOS−/−( n = 10) exhibited a 52% reduction in myocardial injury versus wild-type controls ( P < 0.05). PCR analysis of myocardial inducible NO synthase (iNOS) mRNA levels revealed a significant ( P < 0.05) increase in the UNC eNOS−/− mice compared with wild-type mice, and there was no significant difference between the Har eNOS−/− and wild-type mice. UNC eNOS−/− mice treated with an iNOS inhibitor (1400W) exacerbated the extent of myocardial necrosis. When treated with 1400W, Har eNOS−/− did not exhibit a significant increase in myocardial necrosis. These data demonstrate that two distinct strains of eNOS−/− mice display opposite responses to MI/R. Although the protection seen in the UNC eNOS−/− mouse may result from compensatory increases in iNOS, other genes may be involved.

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Agreement between Impression of Stroke in the Emergency Department and Diagnosis at the Neurology Department

Advances in Bioscience and Clinical Medicine ◽

10.7575/aiac.abcmed.v.7n.3p.7 ◽

2019 ◽

Vol 7 (3) ◽

pp. 7

Author(s):

Samad Shams-Vahdati ◽

Alireza Ala ◽

Eliar Sadeghi-Hokmabad ◽

Neda Parnianfard ◽

Maedeh Gheybi ◽

...

Keyword(s):

Emergency Department ◽

Ischemic Stroke ◽

Statistical Significance ◽

Retrospective Chart Review ◽

Chi Square ◽

Prevention Therapy ◽

Significant Difference ◽

Chi Square Test ◽

Academic Teaching Hospital ◽

The Mean

Background: Missing to detect an ischemic stroke in the emergency department leads to miss acute interventions and treatment with secondary prevention therapy. Our study examined the diagnosis of stroke in the emergency department (ED) and neurology department of an academic teaching hospital. Methods and Materials: A retrospective chart review was performed from March 2017 to March 2018. ED medical document (chart) were reviewed by a stroke neurologist to collect the clinical diagnosis and characteristics of ischemic stroke patients. For determining the cases of misdiagnosed and over diagnosed data, the administrative data codes were compared with the chart adjudicated diagnosis. The adjusted estimate of effect was estimated through testing the significant variables in a multivariable model. The comparisons were done with chi square test. Statistical significance was considered at P < 0.05. Results: Of 861 patients of the study, 54% were males and 43% were females; and the mean age of them was 66.51 ± 15.70. We find no statically significant difference between patient’s Glasgow Coma Scale (GCS) in the emergency department (12.87±3.25) and patients GCS in the neurology department (11.77±5.15). There were 18 (2.2%) overdiagnosed of ischemic stroke, 8 (0.9%) misdiagnosed of ischemic stroke and 36 (4.1%) misdiagnosed of hemorrhagic strokes in the emergency department. Conclusion: There was no significant difference between impression of stroke in the emergency department and diagnosis at the neurology department.

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Nitric oxide synthase-2 (CCTTT)n polymorphism is associated with local gene expression and clinical manifestations in patients with chronic rhinosinusitis

European Journal of Inflammation ◽

10.1177/20587392211052948 ◽

2022 ◽

Vol 20 ◽

pp. 205873922110529

Author(s):

Kota Takemoto ◽

Sachio Takeno ◽

Takashi Ishino ◽

Tsutomu Ueda ◽

Takao Hamamoto ◽

...

Keyword(s):

Nitric Oxide ◽

Chronic Rhinosinusitis ◽

Mrna Level ◽

Clinical Manifestations ◽

Inferior Turbinate ◽

Recurrence Risk ◽

Ethmoid Sinus ◽

Mrna Levels ◽

Repeat Polymorphism ◽

Significant Difference

Introduction Nitric oxide (NO) is synthesized through NO synthase (NOS). The proximal NOS2 gene promoter contains the pentanucleotide CCTTT repeat polymorphism. We examined whether CCTTT repeats are associated with NOS2 expression in the sinonasal tissues and clinical manifestations in patients with chronic rhinosinusitis. Methods Mucosal specimens were obtained from the ethmoid sinus and inferior turbinate of 30 eosinophilic chronic rhinosinusitis (ECRS) and 28 non-ECRS patients. CCTTT repeats were classified into short alleles (S), with less than or equal to 14, and long alleles (L), with more than 14. The subjects were classified into the L/S + L/L and S/S groups. Results In ECRS, the NOS2 mRNA levels of the ethmoid sinus mucosa were significantly higher in the L/S + L/L group than in the S/S group (median, 1.66 and 0.77, respectively). On the ther hand, ECRS patients showed no significant difference in the NOS2 mRNA level of the inferior turbinate between the L/S + L/L group and the S/S group (median, 0.63 and 0.88, respectively). In ECRS, preoperative SNOT-22 were significantly higher in the L/S + L/L group than in the S/S group, whereas the former group showed a lower postoperative recurrence risk. Conclusion CCTTT repeat polymorphism in the NOS2 promotor gene may be a useful indicator to evaluate ECRS severity and prognosis.

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Phenothiazine Inhibits Neuroinflammation and Inflammasome Activation Independent of Hypothermia After Ischemic Stroke

10.21203/rs.3.rs-673718/v1 ◽

2021 ◽

Author(s):

Sichao Guo ◽

Xiaokun Geng ◽

Hangil Lee ◽

Yuchuan Ding

Keyword(s):

Cell Death ◽

Ischemic Stroke ◽

Brain Activity ◽

Ischemia Reperfusion ◽

Inflammasome Activation ◽

Mrna Levels ◽

Sprague Dawley ◽

Neuroinflammatory Response ◽

Protein Levels ◽

Caspase 1

Abstract A depressive or hibernation-like effect of chlorpromazine and promethazine (C + P) on brain activity was reported to induce neuroprotection, with or without induced-hypothermia. However, the underlying mechanisms remain unclear. The current study evaluated the pharmacological function of C + P on the inhibition of neuroinflammatory response and inflammasome activation after ischemia/reperfusion. A total of 72 adult male Sprague-Dawley rats were subjected to 2 h middle cerebral artery occlusion (MCAO) followed by 6 or 24 h reperfusion. At the onset of reperfusion, rats received C + P (8 mg/kg) with temperature control. Brain cell death was detected by measuring CD68 and myeloperoxidase (MPO) levels. Inflammasome activation was measured by mRNA levels of NLRP3, IL-1β, and TXNIP, and protein quantities of NLRP3, IL-1β, TXNIP, cleaved-Caspase-1, and IL-18. Activation of JAK2/STAT3 pathway was detected by the phosphorylation of STAT3 (p-STAT3) and JAK2 (p-JAK2), and the co-localization of p-STAT3 and NLRP3. Activation of the p38 pathway was assessed with the protein levels of p-p38/p38. The mRNA and protein levels of HIF-1α, FoxO1, and p-FoxO1, and the co-localization of p-STAT3 with HIF-1α or FoxO1 were quantitated. As expected, C + P significantly reduced cell death and attenuated the neuroinflammatory response as determined by reduced CD68 and MPO. C + P decreased ischemia-induced inflammasome activation, shown by reduced mRNA and protein expressions of NLRP3, IL-1β, TXNIP, cleaved-Caspase-1, and IL-18. Phosphorylation of JAK2/STAT3 and p38 pathways and the co-localization of p-STAT3 with NLRP3 were also inhibited by C + P. Furthermore, mRNA levels of HIF-1α and FoxO1 were decreased in the C + P group. While C + P inhibited HIF-1α protein expression, it increased FoxO1 phosphorylation, which promoted the exclusion of FoxO1 from the nucleus and inhibited FoxO1 activity. At the same time, C + P reduced the co-localization of p-STAT3 with HIF-1α or FoxO1. In conclusion, C + P treatment conferred neuroprotection in stroke by suppressing neuroinflammation and NLRP3 inflammasome activation. The present study suggests that JAK2/STAT3/p38/HIF-1α/FoxO1 are vital regulators and potential targets for efficacious therapy following ischemic stroke.

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Aligned Expression of IFI16 and STING Genes in RRMS Patients’ Blood

Endocrine Metabolic & Immune Disorders - Drug Targets ◽

10.2174/1871530319666190729112246 ◽

2020 ◽

Vol 20 (6) ◽

pp. 878-886

Author(s):

Sobhan Helbi ◽

Behnam Ravanbakhsh ◽

Mohammad Karimi ◽

Wesam Kooti ◽

Nahid Jivad

Keyword(s):

Mrna Level ◽

Critical Role ◽

Control Group ◽

Case Group ◽

Common Disease ◽

Type I ◽

Mrna Levels ◽

Dna Sensing ◽

Blood Samples ◽

Significant Difference

Objective: Multiple sclerosis (MS) is a chronic neurodegenerative disease of the central nervous system. The most common disease phenotype is Relapsing-Remitting MS (RRMS). Beta interferons are the first line of RRMS patients’ treatment. Interferon-inducible protein 16 (IFI16) as a DNA sensing molecule and its downstream complex stimulator of interferon genes (STING) play a critical role in the activation of type I interferons. Hence we aimed to evaluate the expression rate of IFI16 and STING in RRMS patients’ blood under a different type of IFNβ treatment. Methods: In the present study, 99 individuals participated. The participants were divided into 4 groups: 28 control subjects, 25 new cases of RRMS patients, 25 RRMS patients treated with IFNβ-1a (B1a), 21 RRMS patients treated with IFNβ-1b (B1b). The EDTA-treated blood samples were taken and transferred at standard conditions to the Cellular and Molecular Research Center of Shahrekord University of Medical Sciences, RNA was extracted and converted into cDNA. To evaluate the expression of IFI16 and STING, the Real-Time PCR method using SYBR Green/ROX qPCR master mix was performed done. The level of genes expression was measured using 2–ΔΔCt method. The obtained data were analyzed using SPSS v22 software. Results: Comparison of the IFI and STING mRNA expression in blood samples in association with gender and age showed no significant differences (p>0.05). Also, the evaluation of IFI16 mRNA level revealed that the IFI16 genes’ expressions were remarkably higher in the new case group compared to the control group, however, STING expression did not show any significant difference. The mRNA levels of IFI16 and STING in IFNβ-treated groups were significantly lower than the new case group (p<0.001). Also, the genes’ expressions in both the IFNβ-treated groups were significantly lower compared to the control group (p<0.001). In the assessment of the correlation of IFI16 and STING expressions with age and sex in different research groups, no statistically significant differences were seen (p>0.05). Conclusion: Perhaps the IFNβ therapy decreases the IFI16 and STING expression in a STINGdependent pathway as a negative feedback mechanism for regulation of the immune system and suppression of pro-inflammatory cytokines production. The important role of DNA sensing molecules and STING-dependent pathway in MS gives a new insight into future treatment based on STING-direct therapies.

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Abstract P13: Faster Treatment Speed is Not Associated With Bleeds: Intravenous Alteplase Treatment in the Ultra-Early Time Window is Safe

Stroke ◽

10.1161/str.52.suppl_1.p13 ◽

2021 ◽

Vol 52 (Suppl_1) ◽

Author(s):

Michelle Provencher ◽

Ashley Scherman ◽

Elizabeth Baraban ◽

Robert Jackson ◽

Tamela L Stuchiner ◽

...

Keyword(s):

Ischemic Stroke ◽

Acute Ischemic Stroke ◽

Time Window ◽

Statistical Significance ◽

Hospital Length Of Stay ◽

Bleeding Complications ◽

Stroke Patients ◽

Related Complication ◽

Discharge Disposition ◽

Significant Difference

Previous studies have shown that faster Door to Needle (DTN) treatment times are associated with better outcomes for acute ischemic stroke patients. With the continued push for faster times, we aimed to determine if DTN treatment times in the 30- vs 45-minute time window yielded statistically significant differences in outcomes or complications. Data obtained from a multi-state stroke registry included acute ischemic stroke patients ≥18 years of age discharged between January 2017 and April 2020, and treated with IV alteplase with DTN times between 25-30 or 40-45 minutes. Outcomes were 90-day Modified Rankin Score (mRS) (0-2 vs 3-6), discharge disposition [home or inpatient rehabilitation facility (IRF) vs other location], complications (any treatment-related complication vs none), and hospital length of stay (LOS). Patients with a documented reason for delay or who received thrombectomy were excluded. Outcomes of patients with 25- to 30-minute DTN times were compared to those with 40- to 45-minute DTN times using generalized linear models and multiple linear regression, adjusting for admission NIHSS, age, gender, race/ethnicity, and medical history. Compared to the 20-25 minute group, patients treated in the 40-45 minute window had higher odds of a documented 90-day mRS of 3 or more (Adjusted Odds Ratio (AOR)=1.19, p=0.253, n=201 ) and treatment-related complication (AOR=1.35, p=0.569) and lower odds of discharge to home or IRF (AOR=0.846, p=0.359). There was little difference in LOS (β=-0.008, p=0.847). None of the outcomes reached statistical significance. Administering alteplase in the 25- to 30-minute window is safe and did not result in an increase in bleeding complications. Although faster treatment times trended toward better outcomes, there was no statistically significant difference between the 25-30 and 40-45 minute DTN treatment times.

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Electro-acupuncture on Nei Guan and Bai Hui: How Does It Affect GRP78 and Caspase-12 Gene Expression in Rats with Ischemia-Reperfusion Injury

Journal of Acupuncture and Herbs ◽

10.1515/tcm-2015-0009 ◽

2015 ◽

Vol 3 (1) ◽

pp. 65-69

Author(s):

Jing Shen ◽

Xiao-Ming Lei ◽

Yang Song ◽

Xing Tan ◽

Qin Liu ◽

...

Keyword(s):

Gene Expression ◽

Reperfusion Injury ◽

Mrna Expression ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Statistical Significance ◽

Tunel Staining ◽

Caspase 12 ◽

Significant Difference ◽

Operation Group

Abstract Objective: To observe the effects of electro-acupuncture (EA) on GRP78 and Caspase-12 gene expression in rats with ischemia- reperfusion injury (IRI) by stimulation on Nei Guan (PC6) and Bai Hui (GV20) points, so that to understand whether or not the protective effects of acupuncture is related to endocytoplasmic reticulum (ER) stressapoptosis passage. Methods: 50 rats were randomly assigned to five groups (10 in each group): normal control(A), pseudo-operation(B), operation(C), Edaravone(D) and EA(E). The ischemia/reperfusion model of middle cerebral artery occlusion (MCAO) was established by suture embolic method. TUNEL staining method was employed to measure the apoptosis index of nerve cells in rats. Real-time polymerase chain reaction (RT-PCR) was employed to measure the mRNA expression of GRP78 and Caspase-12. Results: Compared with normal group and pseudo-operation group, the apoptosis indexes and mRNA expression of GRP78 and Caspase-12 in operation group, Edaravone group and EA group were increased, with statistical significance（P＜0.05 or P＜0.01）; compared with operation group, the apoptosis indexes and Caspase-12 mRNA expression in Edaravone group and EA group were decreased（P＜0.05 or P＜0.01）, but GRP78 mRNA expression were increased（P＜0.01）; there were no significant difference between Edaravone group and EA group on the above indexes（P＞0.05). Conclusion: Acupuncture on Nei Guan and Bai Hui points could effectively suppress the nerve cell apoptosis in cerebral ischemia. The underlying mechanism might be related to upregulation of the ERS-protective GRP78 expression and downregulation of apoptosis-promotion Caspase-12 expression.

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Expression of inhibin alpha in adrenocortical tumours reflects the hormonal status of the neoplasm

Journal of Endocrinology ◽

10.1677/joe.0.1650223 ◽

2000 ◽

Vol 165 (2) ◽

pp. 223-229 ◽

Cited By ~ 40

Author(s):

J Arola ◽

J Liu ◽

P Heikkila ◽

V Ilvesmaki ◽

K Salmenkivi ◽

...

Keyword(s):

Mrna Level ◽

Endocrine Function ◽

Alpha Subunit ◽

Mrna Levels ◽

Adrenal Androgen ◽

Median Percentage ◽

Significant Difference ◽

Adrenocortical Tumours ◽

Subunit Expression ◽

Adrenocortical Carcinomas

Inhibins are gonadal glycoprotein hormones whose main endocrine function is to inhibit pituitary FSH secretion. In addition to testes and ovaries, other steroid-producing organs are sites of inhibin alpha subunit expression. To study the role of inhibins in human adrenal gland, we screened a panel of 150 adrenals (10 normal adrenals, 25 adrenocortical hyperplasias, 65 adrenocortical adenomas, 30 adrenocortical carcinomas and 20 phaeochromocytomas) for inhibin alpha expression. mRNA levels of inhibin alpha subunit were studied in 57 samples and all tissues were stained immunohistochemically with an inhibin alpha subunit-specific antibody. Inhibin alpha mRNA was detected in all adrenocortical tissues. Virilizing adenomas possessed a 10-fold higher median inhibin alpha mRNA expression than did normal adrenals. Bilaterally and nodularly hyperplastic adrenals and other than virilizing adrenocortical tumours had their median inhibin alpha mRNA levels close to those of normal adrenals. Immunohistochemically, inhibin alpha subunit was detectable in all normal and hyperplastic adrenals, as well as in 73% of the adrenocortical tumours. However, the percentage of inhibin alpha-positive cells varied greatly in different tumour types. The median percentage of positive cells was 10 in non-functional and Conn's adenomas, 30 in Cushing's adenomas and 75 in virilizing adenomas. In malignant adrenocortical tumours the median percentage of inhibin alpha-immunopositive cells was 20 in non-functional carcinomas, 30 in Conn's carcinomas, 65 in Cushing's carcinomas and 75 in virilizing carcinomas. All phaeochromocytomas were negative for inhibin alpha subunit both at the mRNA level and immunohistochemically. Our data show that inhibin alpha subunit is highly expressed in both normal and neoplastic androgen-producing adrenocortical cells, with less expression in cortisol-producing and hardly any in aldosterone-producing cells. This suggests a specific role for inhibins in the regulation of adrenal androgen production. We did not find any significant difference in inhibin alpha expression between benign and malignant adrenocortical tumours. Thus inhibin alpha gene does not seem to have a tumour suppressor role in human adrenal cortex.

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mRNA Expression of CYP2E1, CYP2A19, CYP1A2, HSD3B, SULT1A1 and SULT2A1 genes in surgically castrated, immunologically castrated, entire male and female pigs and correlation with androstenone, skatole, indole and Improvac-specific antibody levels

Czech Journal of Animal Science ◽

10.17221/159/2018-cjas ◽

2019 ◽

Vol 64 (No. 2) ◽

pp. 89-97

Author(s):

A. Kubešová ◽

K. Šťastný ◽

M. Faldyna ◽

Z. Sládek ◽

I. Steinhauserová ◽

...

Keyword(s):

Gene Expression ◽

Mrna Expression ◽

Mrna Level ◽

Boar Taint ◽

Control Group ◽

Mrna Levels ◽

Large White ◽

Specific Antibodies ◽

Significant Difference ◽

Entire Male

This study aimed to obtain a comprehensive look at the influence of castration on mRNA expression of the genes CYP2E1, CYP1A2, CYP2A19, HSD3B, SULT2A1 and SULT1A1 and their correlation with boar taint compounds (androstenone, skatole and indole) and Improvac-specific antibodies in a Czech commercial hybrid (Large White × Landrace (sow) × Duroc (boar)). Pigs were divided into groups of entire male pigs (NC), pigs castrated surgically (SC), pigs immunologically castrated and slaughtered 8 weeks (IM8) or 15 weeks (IM15) after the second dose of Improvac, and gilts (GI). Hepatic mRNA expression, measured by quantitative real-time polymerase chain reaction, differed significantly between the control group (entire male pigs) and all groups of interest for CYP2E1, CYP1A2 and CYP2A19. The mRNA level of the HSD3B gene differed significantly between the control group and the IM8, IM15 and GI groups. SULT1A1 gene expression was significantly different between the control group and the SC, IM8 and GI. In the case of SULT2A1, a significant difference was observed only between the control group and IM8 pigs. For all genes and treatment groups described above, expression was increased relative to the control. Significant differences for Improvac-specific antibodies between IM8 and IM15 groups were observed, indicating decrease of antibodies over time. Moreover, negative correlations between androstenone and mRNA levels of CYP2A19, CYP2E1 and SULT1A1 suggest that gene expression is suppressed.

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