IDENTIFICATION OF NEW GENES INVOLVED IN THE REGULATION OF YEAST ALCOHOL DEHYDROGENASE II

ABSTRACT Recessive mutations in two negative control elements, CRE1 and CRE2, have been obtained that allow the glucose-repressible alcohol dehydrogenase (ADHII) of yeast to escape repression by glucose. Both the cre1 and cre2 alleles affected ADHII synthesis irrespective of the allele of the positive effector, ADR1. However, for complete derepression of ADHII synthesis, a wild-type ADR1 gene was required. Neither the cre1 nor cre2 alleles affected the expression of several other glucose-repressible enzymes. A third locus, CCR4, was identified by recessive mutations that suppressed the cre1 and cre2 phenotypes. The ccr4 allele blocked the derepression of ADHII and several other glucose-repressible enzymes, indicating that the CCR4 gene is a positive control element. The ccr4 allele had no effect on the repression of ADHII when it was combined with the ADR1-5 c allele, whereas the phenotypically similar ccr1 allele, which partially suppresses ADR1-5 c, did not suppress the cre1 or cre2 phenotype. Complementation studies also indicated that ccr1 and snf1 are allelic. A model of ADHII regulation is proposed in which both ADR1 and CCR4 are required for ADHII expression. CRE1 and CRE2 negatively control CCR4, whereas CCR1 is required for ADR1 function.

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Ranitidine as an alcohol dehydrogenase inhibitor in acute methanol toxicity in rats

Human & Experimental Toxicology ◽

10.1177/0960327109353777 ◽

2009 ◽

Vol 29 (2) ◽

pp. 93-101 ◽

Cited By ~ 8

Author(s):

Amal A El-Bakary ◽

Sahar A El-Dakrory ◽

Sohayla M Attalla ◽

Nawal A Hasanein ◽

Hala A Malek

Keyword(s):

Alcohol Dehydrogenase ◽

High Performance ◽

Negative Control Group ◽

Positive Control ◽

Negative Control ◽

Control Group ◽

Sprague Dawley ◽

Blood Samples ◽

Methanol Poisoning ◽

Sprague Dawley Rats

Methanol poisoning is a hazardous intoxication characterized by visual impairment and formic acidemia. The therapy for methanol poisoning is alcohol dehydrogenase (ADH) inhibitors to prevent formate accumulation. Ranitidine has been considered to be an inhibitor of both gastric alcohol and hepatic aldehyde dehydrogenase enzymes. This study aimed at testing ranitidine as an antidote for methanol acute toxicity and comparing it with ethanol and 4-methyl pyrazole (4-MP). This study was conducted on 48 Sprague-Dawley rats, divided into 6 groups, with 8 rats in each group (one negative control group [C1], two positive control groups [C2, C3] and three test groups [1, 2 and 3]). C2, C3 and all test groups were exposed to nitrous oxide by inhalation, then, C3 group was given methanol (3 g/kg orally). The three test groups 1, 2 and 3 were given ethanol (0.5 g/kg orally), 4-MP (15 mg/kg intraperitoneally) and ranitidine (30 mg/kg intraperitoneally), respectively, 4 hours after giving methanol. Rats were sacrificed and heparinized, cardiac blood samples were collected for blood pH and bicarbonate. Non-heparinized blood samples were collected for formate levels by high performance liquid chromatography. Eye balls were enucleated for histological examination of the retina. Ranitidine corrected metabolic acidosis (p = .025), decreased formate levels (p = .014) and improved the histological findings in the retina induced by acute methanol toxicity.

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Binding of coenzymes to yeast alcohol dehydrogenase

Journal of the Serbian Chemical Society ◽

10.2298/jsc1002185l ◽

2010 ◽

Vol 75 (2) ◽

pp. 185-194 ◽

Cited By ~ 5

Author(s):

Vladimir Leskovac ◽

Svetlana Trivic ◽

Draginja Pericin ◽

Mira Popovic ◽

Julijan Kandrac

Keyword(s):

Alcohol Dehydrogenase ◽

Equilibrium Constants ◽

Point Mutations ◽

Free Energies ◽

Wild Type ◽

Wild Type Enzyme ◽

Mutant Type ◽

Yeast Alcohol Dehydrogenase ◽

Internal Equilibrium ◽

Individual Reaction

In this work, the binding of coenzymes to yeast alcohol dehydrogenase (EC 1.1.1.1) were investigated. The main criterions were the change in the standard free energies for individual reaction steps, the internal equilibrium constants and the overall changes in the reaction free energies. The calculations were performed for the wild type enzyme at pH 6-9 and for 15 different mutant type enzymes, with single or double point mutations, at pH 7.3. The abundance of theoretical and experimental data enabled the binding of coenzymes to enzyme to be assessed in depth.

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Expression of Ki67 in Precancerous Squamous Cell Skin Lesion of Mice Induced by DMBA

Indonesian Journal of Cancer ◽

10.33371/ijoc.v13i2.638 ◽

2019 ◽

Vol 13 (2) ◽

pp. 44

Author(s):

Siti Nurkasanah ◽

Aida S D Hoemardani ◽

Evlina Suzanna Sinuraya

Keyword(s):

Skin Lesion ◽

Squamous Cell ◽

Precancerous Lesion ◽

Positive Control ◽

Negative Control ◽

Cumulative Number ◽

Wild Type ◽

Imiquimod Cream ◽

Ki67 Expression ◽

One Way Anova

Background: Squamous cell carcinoma (SCC) is an invasive skin cancer that is found in the human skin. SCC growth from the precancerous lesion is known as a wart or papilloma. Ki67 is a protein expressed by cells during proliferation. Cells with higher Ki67 expression showed abnormal regulation of apoptosis. 5-FU is an antimetabolite compound that works as a DNA/RNA pyrimidine antagonist molecule that can induce cell apoptosis. The main objective of this research is to evaluate the inhibition proliferation of precancerous squamous cell in skin lesion induced by DMBA/croton oil treated by using 5-FU cream topically compared to 5% imiquimod cream. Methods: This research assesses three different concentrations of 5-FU cream namely 1%, 2%, and 5% topically on 24 wild type mice divided into 6 groups of 4 each including positive control (with carcinogenesis but without treatment), negative control (without treatment ; normal), carcinogenesis with treatment 5-FU cream (1%, 2%, and 5%) or 5% imiquimod cream. The expression of Ki67 was analyzed using immunohistochemistry. Statistical analysis was performed by one-way ANOVA using SPSS version 23. Results: Based on this research, 5-FU cream treatment during 4 weeks (week 10-week 14) showed the decrease of cumulative number of papilloma (p<0.05) and immunohistochemistry analysis resulted in the lowest Ki67 expression in 5-FU 2% and 5% groups compared to 5% imiquimod cream (p<0.05). Conclusions: 5-FU cream treatment during 4 weeks showed a decrease of papilloma and Ki67 expression in 5-FU cream 2% and

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Positive Control of Swarming, Rhamnolipid Synthesis, and Lipase Production by the Posttranscriptional RsmA/RsmZ System in Pseudomonas aeruginosa PAO1

Journal of Bacteriology ◽

10.1128/jb.186.10.2936-2945.2004 ◽

2004 ◽

Vol 186 (10) ◽

pp. 2936-2945 ◽

Cited By ~ 184

Author(s):

Karin Heurlier ◽

Faye Williams ◽

Stephan Heeb ◽

Corinne Dormond ◽

Gabriella Pessi ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Target Genes ◽

Rna Binding ◽

Regulatory Protein ◽

Lipase Production ◽

Positive Control ◽

Negative Control ◽

Control Element ◽

Signal Molecules ◽

Positive Effects

ABSTRACT In Pseudomonas aeruginosa, the small RNA-binding, regulatory protein RsmA is a negative control element in the formation of several extracellular products (e.g., pyocyanin, hydrogen cyanide, PA-IL lectin) as well as in the production of N-acylhomoserine lactone quorum-sensing signal molecules. RsmA was found to control positively the ability to swarm and to produce extracellular rhamnolipids and lipase, i.e., functions contributing to niche colonization by P. aeruginosa. An rsmA null mutant was entirely devoid of swarming but produced detectable amounts of rhamnolipids, suggesting that factors in addition to rhamnolipids influence the swarming ability of P. aeruginosa. A small regulatory RNA, rsmZ, which antagonized the effects of RsmA, was identified in P. aeruginosa. Expression of the rsmZ gene was dependent on both the global regulator GacA and RsmA, increased with cell density, and was subject to negative autoregulation. Overexpression of rsmZ and a null mutation in rsmA resulted in quantitatively similar, negative or positive effects on target genes, in agreement with a model that postulates titration of RsmA protein by RsmZ RNA.

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REPRESSOR AND ANTIREPRESSOR IN THE REGULATION OF STAPHYLOCOCCAL PENICILLINASE SYNTHESIS

Genetics ◽

10.1093/genetics/75.1.1 ◽

1973 ◽

Vol 75 (1) ◽

pp. 1-17

Author(s):

John Imsande

Keyword(s):

Staphylococcus Aureus ◽

Binding Site ◽

Regulatory Protein ◽

Positive Control ◽

Negative Control ◽

Biochemical Data ◽

Wild Type ◽

Data Support ◽

Operator Binding ◽

Effector Binding

ABSTRACT 5-methyltryptophan (5MT) induces penicillinase synthesis in Staphylococcus aureus. The analog is incorporated into protein by both wild-type and tryptophan-starved cells. Since normal penicillinase repressor appears to contain tryptophan even though penicillinase itself does not, it is concluded that 5MT induces penicillinase synthesis by becoming incorporated into the penicillinase repressor and thereby inactivating the repressor. Thus biochemical data support the existence of a penicillinase repressor and indicate that penicillinase synthesis is regulated by negative control and not by positive control.—In the absence of exogenous tryptophan, staphylococcal penicillinase induction can be inhibited by 7-azatryptophan (7azaT). Because 7azaT is incorporated into protein by tryptophan-starved cells, it is concluded that 7azaT blocks penicillinase induction by inactivating a penicillinase regulatory protein into which the analog has been incorporated. Incorporation of 7azaT does not appear to inactivate the operator binding site or the effector binding site on the penicillinase repressor. Therefore, it appears that 7azaT blocks penicillinase induction by inactivating the penicillinase antirepressor, a protein required for inactivation of the penicillinase repressor and, hence, required for penicillinase induction.

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New SNF genes, GAL11 and GRR1 affect SUC2 expression in Saccharomyces cerevisiae.

Genetics ◽

10.1093/genetics/129.3.675 ◽

1991 ◽

Vol 129 (3) ◽

pp. 675-684 ◽

Cited By ~ 10

Author(s):

L G Vallier ◽

M Carlson

Keyword(s):

Saccharomyces Cerevisiae ◽

Null Mutation ◽

Wild Type ◽

Glucose Limitation ◽

Wild Type Level ◽

Recessive Mutations ◽

New Genes ◽

Complementation Groups ◽

Invertase Gene ◽

Inducible Gene

Abstract To identify new genes required for depression of the SUC2 (invertase) gene in Saccharomyces cerevisiae, we have isolated mutants with defects in raffinose utilization. In addition to mutations in SUC2 and previously identified SNF genes, we recovered recessive mutations that define four new complementation groups, designated snf7 through snf10. These mutations cause defects in the derepression of SUC2 in response to glucose limitation. We also recovered five alleles of gal11 and showed that a gal11 null mutation decreases SUC2 expression to 30% of the wild-type level. Finally, one of the mutants carries a grr1 allele that converts SUC2 from a glucose-inducible gene.

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A mutational analysis of killer toxin resistance in Saccharomyces cerevisiae identifies new genes involved in cell wall (1-->6)-beta-glucan synthesis.

Genetics ◽

10.1093/genetics/133.4.837 ◽

1993 ◽

Vol 133 (4) ◽

pp. 837-849 ◽

Cited By ~ 5

Author(s):

J L Brown ◽

Z Kossaczka ◽

B Jiang ◽

H Bussey

Keyword(s):

Cell Wall ◽

Mutational Analysis ◽

Killer Toxin ◽

Similar Proportion ◽

Wild Type ◽

Beta Glucan ◽

Toxin Resistance ◽

Recessive Mutations ◽

New Genes ◽

Glucan Synthesis

Abstract Recessive mutations leading to killer resistance identify the KRE9, KRE10 and KRE11 genes. Mutations in both the KRE9 and KRE11 genes lead to reduced levels of (1-->6)-beta-glucan in the yeast cell wall. The KRE11 gene encodes a putative 63-kD cytoplasmic protein, and disruption of the KRE11 locus leads to a 50% reduced level of cell wall (1-->6)-glucan. Structural analysis of the (1-->6)-beta-glucan remaining in a kre11 mutant indicates a polymer smaller in size than wild type, but containing a similar proportion of (1-->6)- and (1-->3)-linkages. Genetic interactions among cells harboring mutations at the KRE11, KRE6 and KRE1 loci indicate lethality of kre11 kre6 double mutants and that kre11 is epistatic to kre1, with both gene products required to produce the mature glucan polymer at wild-type levels. Analysis of these KRE genes should extend knowledge of the beta-glucan biosynthetic pathway, and of cell wall synthesis in yeast.

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Aktivitas Antifungi Air Perasan Bawang Merah (Allium ascalonicum L.) Terhadap Candida albicans Secara In Vitro

Jurnal Ilmu Kedokteran ◽

10.26891/jik.v9i2.2015.71-77 ◽

2017 ◽

Vol 9 (2) ◽

pp. 71

Author(s):

Nurhasanah Nurhasanah ◽

Fauzia Andrini ◽

Yulis Hamidy

Keyword(s):

Candida Albicans ◽

Antifungal Activity ◽

Allium Sativum ◽

Fungicidal Activity ◽

Positive Control ◽

Negative Control ◽

Randomized Design ◽

Significant Difference ◽

Completely Randomized Design

Shallot (Allium ascalonicum L.) has been known as traditional medicine. Shallot which has same genus with garlic(Allium sativum L.) contains allicin that is also found in garlic and has been suspected has fungicidal activity toCandida albicans. It is supported by several researches. Therefore, shallot is suspected has antifungal activity too.The aim of this research was to know antifungal activity of shallot’s water extortion againsts Candida albicans invitro. This was a laboratory experimental research which used completely randomized design, with diffusion method.Shallot’s water extortion was devided into three concentrations, there were 50%, 100% and 200%. Ketoconazole 2%was positive control and aquadest was negative control. The result of this research based on analysis of varians(Anova), there was significant difference between several treatments and was confirmed with Duncan New MultipleRange Test (DNMRT) p<0,05, there was significant difference between 100% shallot’s water extortion with othertreatments, but there was no significant difference between 50% shallot’s water extortion with 200% shallot’s. Theconclusion was shallot’s water extortion had antifungal activity againsts Candida albicans with the best concentration100%, but it was lower than ketoconazole 2%.

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The Comparison of Ostecyte in the Pressure area and Tension area on Tooth Movement Because of Hyperbaric Oxygen Therapy

DENTA ◽

10.30649/denta.v12i1.162 ◽

2018 ◽

Vol 12 (1) ◽

pp. 34

Author(s):

Arya Barahmanta ◽

Muhammad Faizal Winaris ◽

Pambudi Raharjo

Keyword(s):

Hyperbaric Oxygen ◽

Oxygen Therapy ◽

Tooth Movement ◽

Bone Remodelling ◽

Orthodontic Tooth Movement ◽

Negative Control Group ◽

Positive Control ◽

Negative Control ◽

Control Group ◽

Pressure Area

Background: Orthodontic tooth movement is a interaction prosess of resorption and deposition of bone remodeling. Orthodontic tooth movement by mechanical strength causes changes in alveolar bone. Osteocyte is an essential cell to respond bone remodelling. Hyperbaric Oxygen Therapy affects production of osteocyte because it can release Reactive Oxygen Species (ROS) and Nitrid Oxide (NO). Purpose: To determine the difference number of osteocyte in pressure and tension area during tooth movement by adjuvant of Hyperbaric Oxygen 2,4 ATA during 7 days starting on day 8 to day 14. Materials and Methods: This research used Completery Randomized Control Group Post Test Only Design. 36 cavia cobaya (male) were divided into 3 groups randomly : the negative control groups, positive control group, and treatment group. Preparat staining used Hematoxylin Eosin (HE) and calculated on microscop 1000x with 20 field of view. Data analyses used one way ANOVA and LSD test then compared each area by using paired T test. Result: The data showed that the treatment group (P=10,67) tension area has the highest number of osteocyte than negative control group (K-=3,67), positive control (K+=7,42). In the pressure area showed that negative control group (K-=5,00) has the highest than positive control group (K+=3,83) and treatment (P=3,25). Conclusion: Therapy HBO 2,4 ATA 7 days starting on day 8 to day 14 is could increase osteocyte in the tissue to stimulate process of bone remodelling.<pre> </pre>Keywords: Hyperbaric Oxygen, Tooth movement, Bone remodeling, Osteocyte Correspondence: Arya Brahmanta, Department of Orthodonty, Faculty of Dentistry, Hang Tuah University, Arif Rahman Hakim 150, Surabaya, Phone 031-5945864, Email: <a href="mailto:[email protected]">arya.brahmanta@hangtuah.ac.id</a>

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UJI AKTIVITAS HEPATOPROTEKTOR KOMBINASI EKSTRAK ETANOL DAUN PANDAN WANGI (Pandanus amaryllifolius roxb) DAN KAYU MANIS (Cinnamomum burmanii) TERHADAP KADAR MDA YANG DIINDUKSI PARASETAMOL

Jurnal Kesehatan Kusuma Husada ◽

10.34035/jk.v12i1.580 ◽

2020 ◽

pp. 68-73

Author(s):

Yuni Asri Mulatsih Agami ◽

Eka Wisnu Kusuma

Keyword(s):

Oxidative Stress ◽

Ethanol Extract ◽

Optimal Dose ◽

Experimental Methods ◽

Positive Control ◽

Negative Control ◽

Male Rats ◽

Negative Group ◽

Ic50 Value ◽

Dose Combination

Kasus penyakit hati semakin meningkat seiring penggunaan senyawa hepatotoksin salah satunya karena penggunaan parasetamol dengan dosis berlebih. Hal tersebut dapat meningkatkan produksi radikal bebas sehingga memicu terjadinya stress oksidatif yang dapat menimbulkan kerusakan jaringan yang ditandai dengan peningkatan kadar Malondialdehyde (MDA). Stress oksidatif dapat diatasi dengan antioksidan dari berbagai tanaman. Kulit kayu manis memiliki aktivitas antioksidan dengan nilai IC50 53ppm dan daun pandan wangi 39,7% Penelitian ini bertujuan untuk mengetahui aktivitas kombinasi ekstrak etanol daun pandan wangi dan kayu manis dalam menurunkan kadar MDA. tikus yang diinduksi parasetamol. Penelitian menggunakan metode eksperimental, dilakukan selama 9 hari dengan 30 ekor tikus jantan dibagi menjadi 6 Kelompok, yaitu: Normal diberi aquadest, Kontrol Positif diberi silimarin 100 mg/kgBB, Kontrol Negatif diberi CMC-Na 0,05%, serta 3 kelompok lainnya diberi kombinasi ekstrak daun pandan wangi:kayu manis berturut-turut dosis I (25:75), dosis II (50:50), dosis III (75:25). Semua kelompok diinduksi parasetamol 2,5 g/kgBB pada hari ke-7 setelah 30 menit perlakuan, kecuali kelompok normal. Pada hari ke 9 dilakukan pengukuran kadar MDA dengan metode TBARs menggunakan spektrofotometri. Pemberian kombinasi ekstrak etanol daun pandan wangi dan kayu manis dapat menurunkan kadar MDA dengan kombinasi dosis yang paling optimal adalah 75:25 berdasarkan statistik dengan nilai signifikan 0,000<0,05 dibandingkan dengan kelompok negatif. Cases of liver disease have increased with the use of hepatotoxin compounds, one of which is due to the use of paracetamol with excessive doses. This can increase the production of free radicals so that it triggers oxidative stress which can cause tissue damage which is characterized by increased levels of Malondialdehyde (MDA). Oxidative stress can be overcome with antioxidants from various plants. Cinnamomum burmanii has antioxidant activity with IC50 value of 53ppm and Pandanus amarrylifolius 39.7%. This study aims to determine the combined activity of ethanol extract of Pandanus amarrylifolius and Cinnamomum burmanii in reducing MDA levels. Paracetamol-induced rats. Research using experimental methods, conducted for 9 days with 30 male rats divided into 6 groups, namely: Normal given aquadest, Positive Control were given silimarin 100 mg / kgBB, Negative Control was given CMC-Na 0.05%, and 3 other groups were given a combination of Pandanus amarrylifolius extract: Cinnamomum burmanii dose I (25:75), dose II (50:50), dose III (75:25). All groups induced paracetamol 2.5 g / kgBB on the 7th day after 30 minutes of treatment, except the normal group. On the 9th day MDA levels were measured using the TBARs method using spectrophotometry. Giving a combination of Pandanus amarrylifolius and Cinnamomum burmanii ethanol extract can reduce MDA levels with the most optimal dose combination is 75:25 based on statistics with a significant value of 0,000<0.05 compared with the negative group.

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