• Title/Summary/Keyword: Single molecules

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ROLE OF REACTIVE OXYGEN SPECIES IN MALE INFERTILITY

  • Sharma, Rakesh K.;Agarwal, Ashok
    • 대한생식의학회:학술대회논문집
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    • 2000.06a
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    • pp.13-28
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    • 2000
  • Human spermatozoa exhibit a capacity to generate ROS and initiate peroxidation of the unsaturated fatty acids in the sperm plasma membrane, which plays a key role in the etiology of male infertility. The short half-life and limited diffusion of these molecules is consistent with their physiologic role in key biological events such as acrosome reaction and hyperactivation. The intrinsic reactivity of these metabolites in peroxidative damage induced by ROS, particularly $H_2O_2$ and the superoxide anion, has been proposed as a major cause of defective sperm function in cases of male infertility. The number of antioxidants known to attack different stages of peroxidative damage is growing, and it will be of interest to compare alpha-tocopherol and ascorbic acid with these for their therapeutic potential in vitro and in vivo. Both spermatozoa and leukocytes generate ROS, although leukocytes produce much higher levels. The clinical significance of leukocyte presence in semen is controversial. Seminal plasma confers some protection against ROS damage because it contains enzymes that scavenge ROS, such as catalase and superoxide dismutase. A variety of defense mechanisms comprising a number of antioxidants can be employed to reduce or overcome oxidative stress caused by excessive ROS. Determination of male infertility etiology is important, as it will help us develop effective therapies to overcome excessive ROS generation. ROS can have both beneficial and detrimental effects on the spermatozoa and the balancing between the amounts of ROS produced and the amounts scavenged at any moment will determine whether a given sperm function will be promoted or jeopardized. Accurate assessment of ROS levels and, subsequently, OS is Vital, as this will help clinicians both elucidate the fertility status and identify the subgroups of patients that respond or do not respond to these therapeutic strategies. The overt commercial claims of antioxidant benefits and supplements for fertility purposes must be cautiously looked into, until proper multicentered clinical trials are studied. From the current data it appears that no Single adjuvant will be able to enhance the fertilizing capacity of sperm in infertile men, and a combination of the possible strategies that are not toxic at the dosage used would be a feasible approach.

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Regulation of Gene Expression and 3-Dimensional Structure of DNA (유전자 발현 조절과 DNA 3차원적 구조와의 관계)

  • 김병동
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.149-155
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    • 1987
  • Growth and development of a higher plant, or any living organism for that matter, could be defined as an orderly expression of the genome in time and space in close interaction with the environment. During differentiation and development of a tissue or organ a group of genes must be selectively turned on or turned off mainly by trans-acting regulators. In this general concept of regulation of regulation of gene expression, a DNA molecule is recognized at a specific nucleotide sequence by DNA-binding factors. Molecular biology of the regulatory factors such as hormones, and their receptors, target DNA sequences and DNA-binding proteins are well advanced. What is not clearly understood is the molecular basis of the interactions between DNA and binding factors, expecially of the usages of the dyad symmetry of the target DNA sequences and the dimeric nature of the DNA-binding proteins. A unique 3-dimensional structure of DNA has been proposed that may play an important role in the orderly expression of the gene. A foldback intercoil (FBI) DNA configuration which was originally found by electron microscopy among mtDNA molecules from pearl millet has some unique features. The FBI configuration of DNA is believed to be formed when a flexible double helix folds back and interwines in the widened major grooves resulting in a four stranded, intercoil DNA whose thickness is the same as that of double stranded DNA. More recently, the FBI structure of DNA has been also induced in vitro by a novel enzyme which was purified from pearl millet mitochondria. It has been proposed that the FBI DNA could be utillized in intramolecular recombination which leads to inversion or deletion, and in intermolecular recombination which can lead to either site-specific recombination, genetic recombination via single strand invasion, or cross strand recombination. The structure and function of DNA in 3-dimensional aspect is emphasized for better understanding orderly expression of genes during growth and development.

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The Structures of Dehydrated Fully $Ca^{2+}$-Exchanged Zeolite A and of Its iodine Sorption Complex (칼슘 이온으로 완전히 치환된 제올라이트 A의 탈수 구조와 칼슘 치환 제올라이트 A에 요오드가 흡착된 착물 구조)

  • Jang, Se-Bok;Han, Yong-Wook;Kim, Duk-Soo;Kim, Yang
    • Korean Journal of Crystallography
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    • v.1 no.2
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    • pp.76-83
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    • 1990
  • The structures of dehydrated fully Ca2+ _exchanged zeolite A (a: 12.2a3(2) A and of its iodine sorption complex (a=12.258(2) A) have been determined by single-crystal X-ray diffraction methods in the cubic space group. Pm3m at 21(1)℃.the structures were refined to final R(weighted) indices of 0.081 with 206 reflections and 0.084 with 173 reflections, respectively for which I>3 w (I). In each structure. six divalent cations are located on three different theefold axes associated with 6-ring oxygens. Dehydrated Ca2+_A sorbs ca. 6.0 diiodine molecules per unit cell at 80℃(vapor pressure of I: is ca. 14.3 Torr). Each iodine molecule makes a close approach. along its ahs to framework oxygen atom with I-I distance of 2.71(2) A, I-O distance of 3.32(3) k. and I-I-0=180℃. The result indicates that diiodine molecule forms charge transfer complex with framework oxygen.

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Effects of Recombinant Imperatoxin A (IpTxa) Mutants on the Rabbit Ryanodine Receptor

  • Seo, In-Ra;Choi, Mu-Rim;Park, Chul-Seung;Kim, Do Han
    • Molecules and Cells
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    • v.22 no.3
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    • pp.328-335
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    • 2006
  • Imperatoxin A ($IpTx_a$), a 3.7 kDa peptide from the African scorpion Pandinus imperator, is an agonist of the skeletal muscle ryanodine receptor (RyR1). In order to study the structure of the toxin and its effect on RyR1, $IpTx_a$ cDNA was PCR-amplified using 3 pairs of primers, and the toxin was expressed in E. coli. The toxin was further purified by chromatography, and various point mutants in which basic amino acids were substituted by alanine were prepared by site-directed mutagenesis. Studies of single channel properties by the planar lipid bilayer method showed that the recombinant $IpTx_a$ was identical to the synthetic $IpTx_a$ with respect to high-performance liquid chromatography mobility, amino acid composition and specific effects on RyR1. Mutations of certain basic amino acids ($Lys^{19}$, $Arg^{23}$, and $Arg^{33}$) dramatically reduced the capacity of the peptide to activate RyRs. A subconductance state predominated when $Lys^8$ was substituted with alanine. These results suggest that some basic amino acid residues in $IpTx_a$ are important for activation of RyR1, and that $Lys^8$ plays an important role in regulating the gating mode of RyR1.

Structural and Mechanistic Insights into the Tropism of Epstein-Barr Virus

  • Mohl, Britta S.;Chen, Jia;Sathiyamoorthy, Karthik;Jardetzky, Theodore S.;Longnecker, Richard
    • Molecules and Cells
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    • v.39 no.4
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    • pp.286-291
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    • 2016
  • Epstein-Barr virus (EBV) is the prototypical ${\gamma}$-herpesvirus and an obligate human pathogen that infects mainly epithelial cells and B cells, which can result in malignancies. EBV infects these target cells by fusing with the viral and cellular lipid bilayer membranes using multiple viral factors and host receptor(s) thus exhibiting a unique complexity in its entry machinery. To enter epithelial cells, EBV requires minimally the conserved core fusion machinery comprised of the glycoproteins gH/gL acting as the receptor-binding complex and gB as the fusogen. EBV can enter B cells using gp42, which binds tightly to gH/gL and interacts with host HLA class II, activating fusion. Previously, we published the individual crystal structures of EBV entry factors, such as gH/gL and gp42, the EBV/host receptor complex, gp42/HLA-DR1, and the fusion protein EBV gB in a postfusion conformation, which allowed us to identify structural determinants and regions critical for receptor-binding and membrane fusion. Recently, we reported different low resolution models of the EBV B cell entry triggering complex (gHgL/gp42/HLA class II) in "open" and "closed" states based on negative-stain single particle electron microscopy, which provide further mechanistic insights. This review summarizes the current knowledge of these key players in EBV entry and how their structures impact receptor-binding and the triggering of gB-mediated fusion.

Concurrent Hypermethylation of SFRP2 and DKK2 Activates the Wnt/β-Catenin Pathway and Is Associated with Poor Prognosis in Patients with Gastric Cancer

  • Wang, Hao;Duan, Xiang-Long;Qi, Xiao-Li;Meng, Lei;Xu, Yi-Song;Wu, Tong;Dai, Peng-Gao
    • Molecules and Cells
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    • v.40 no.1
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    • pp.45-53
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    • 2017
  • Aberrant hypermethylation of Wnt antagonists has been observed in gastric cancer. A number of studies have focused on the hypermethylation of a single Wnt antagonist and its role in regulating the activation of signaling. However, how the Wnt antagonists interacted to regulate the signaling pathway has not been reported. In the present study, we systematically investigated the methylation of some Wnt antagonist genes (SFRP2, SFRP4, SFRP5, DKK1, DKK2, and APC) and their regulatory role in carcinogenesis. We found that aberrant promoter methylation of SFRP2, SFRP4, DKK1, and DKK2 was significantly increased in gastric cancer. Moreover, concurrent hypermethylation of SFRP2 and DKK2 was observed in gastric cancer and this was significantly associated with increased expression of ${\beta}-catenin$, indicating that the joint inactivation of these two genes promoted the activation of the Wnt signaling pathway. Further analysis using a multivariate Cox proportional hazards model showed that DKK2 methylation was an independent prognostic factor for poor overall survival, and the predictive value was markedly enhanced when the combined methylation status of SFRP2 and DKK2 was considered. In addition, the methylation level of SFRP4 and DKK2 was correlated with the patient's age and tumor differentiation, respectively. In conclusion, epigenetic silencing of Wnt antagonists was associated with gastric carcinogenesis, and concurrent hypermethylation of SFRP2 and DKK2 could be a potential marker for a prognosis of poor overall survival.

Properties of the Microinterface formed by Phosphatidylcholine and 1-Butanol as Reaction Media of Hydrolysis of Phosphatidylcholine

  • Yamazaki, Keiju;Imai, Masanao;Suzuki, Isao
    • Proceedings of the Membrane Society of Korea Conference
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    • 2004.05a
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    • pp.82-85
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    • 2004
  • Microinterface of W/Omicroemulsion prepared by phosphatidylcholine was used as reaction media of hydrolysis of phosphatidylcholine by phospholipaseA$_2$. Phosphatidylcholine was used as an amphiphile and was acted as a substrate. Organic phase of W/Omicroemulsion in this study was prepared by mixed organic solvents i.e. 2,2,4-trimethylpentane (isooctane) as a main solvent and 1-butanol as a co-solvent. The effect of added 1-butanol was remarkable not only on reaction beginning but also on high reaction rate. The hydrolysis reaction was dramatically initiated when 1-butanol was injected into the running isooctane/PC system. The enhancement by 1-butanol addition into single organic solvent was our original finding compare with previous conventional organic solvent. The reaction rate was elevated by the added amount of 1-butanol. The enhanced reaction rate was about 150-folds. This enhancement was speculated as 1-butanol adsorption on the microinterface. The adsorbed 1-butanol improved the properties of microinterface, especially its mobility was increased by difference of the chain length between phosphatidylcholine and 1-butanol. PhospholipaseA$_2$ molecules were located on the microinterface due to modified mobility of microinterface. Located phospholipaseA$_2$ on the microinterface reacted easily with phosphatidylcholine molecule. As a result high reaction rate was obtained. Microinterfacial properties were successfully improved by adsorbed 1-butanol molecule, and were favorable to appear higher reactivity of phospholipaseA$_2$.

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Metagenomic SMRT Sequencing-Based Exploration of Novel Lignocellulose-Degrading Capability in Wood Detritus from Torreya nucifera in Bija Forest on Jeju Island

  • Oh, Han Na;Lee, Tae Kwon;Park, Jae Wan;No, Jee Hyun;Kim, Dockyu;Sul, Woo Jun
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1670-1680
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    • 2017
  • Lignocellulose, composed mostly of cellulose, hemicellulose, and lignin generated through secondary growth of woody plant, is considered as promising resources for biofuel. In order to use lignocellulose as a biofuel, biodegradation besides high-cost chemical treatments were applied, but knowledge on the decomposition of lignocellulose occurring in a natural environment is insufficient. We analyzed the 16S rRNA gene and metagenome to understand how the lignocellulose is decomposed naturally in decayed Torreya nucifera (L) of Bija forest (Bijarim) in Gotjawal, an ecologically distinct environment. A total of 464,360 reads were obtained from 16S rRNA gene sequencing, representing diverse phyla; Proteobacteria (51%), Bacteroidetes (11%) and Actinobacteria (10%). The metagenome analysis using single molecules real-time sequencing revealed that the assembled contigs determined originated from Proteobacteria (58%) and Actinobacteria (10.3%). Carbohydrate Active enZYmes (CAZy)- and Protein families (Pfam)-based analysis showed that Proteobacteria was involved in degrading whole lignocellulose, and Actinobacteria played a role only in a part of hemicellulose degradation. Combining these results, it suggested that Proteobacteria and Actinobacteria had selective biodegradation potential for different lignocellulose substrates. Thus, it is considered that understanding of the systemic microbial degradation pathways may be a useful strategy for recycle of lignocellulosic biomass, and the microbial enzymes in Bija forest can be useful natural resources in industrial processes.

Synthesis and Optical Properties of TiO2/TiOF2 Composite Powder with Controlled Phase Fractions via an Ultrasonic Spray Pyrolysis Process (초음파 분무 열분해 공정을 이용한 TiO2와 TiOF2 복합체 분말의 합성과 상 분율에 따른 광학적 성질)

  • Hwangbo, Young;Park, Woo-Young;Lee, Young-In
    • Korean Journal of Materials Research
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    • v.27 no.6
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    • pp.325-330
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    • 2017
  • Photoelectron-hole separation efficiency plays an important role in the enhancement of the photocatalytic activity of photocatalysts towards the degradation of organic molecules. In this study, $TiO_2/TiOF_2$ heterostructured composite powders with suitable band structures, which structures are able to separate photoelectron-hole pairs, have been synthesized using a simple and versatile ultrasonic spray pyrolysis process. In addition, their phase volume fractions have been controlled by varying the pyrolysis temperature from $400^{\circ}C$ to $800^{\circ}C$. The structural and optical properties of the synthesized powders have been characterized by X-ray diffraction, scanning electronic microscopy and UV-vis spectroscopy. The powder with a phase volume ratio close to 1, compared with single $TiOF_2$ and other composite powders with different phase volume fractions, was found to have superior photocatalytic activity for the degradation of rhodamine B. This result shows that the $TiO_2/TiOF_2$ heterostructure promotes the separation of the photoinduced electrons and holes and that this powder can be applicable to environmental cleaning applications.

Biflorin Ameliorates Memory Impairments Induced by Cholinergic Blockade in Mice

  • Jeon, Se Jin;Kim, Boseong;Ryu, Byeol;Kim, Eunji;Lee, Sunhee;Jang, Dae Sik;Ryu, Jong Hoon
    • Biomolecules & Therapeutics
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    • v.25 no.3
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    • pp.249-258
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    • 2017
  • To examine the effect of biflorin, a component of Syzygium aromaticum, on memory deficit, we introduced a scopolamine-induced cognitive deficit mouse model. A single administration of biflorin increased latency time in the passive avoidance task, ameliorated alternation behavior in the Y-maze, and increased exploration time in the Morris water maze task, indicating the improvement of cognitive behaviors against cholinergic dysfunction. The biflorin-induced reverse of latency in the scopolamine-treated group was attenuated by MK-801, an NMDA receptor antagonist. Biflorin also enhanced cognitive function in a naïve mouse model. To understand the mechanism of biflorin for memory amelioration, we performed Western blot. Biflorin increased the activation of protein kinase C-${\zeta}$ and its downstream signaling molecules in the hippocampus. These results suggest that biflorin ameliorates drug-induced memory impairment by modulation of protein kinase C-${\zeta}$ signaling in mice, implying that biflorin could function as a possible therapeutic agent for the treatment of cognitive problems.