Fourth Domain Research Articles

Distinctive Properties of Dark Reversion Kinetics between Two Red/Green-Type Cyanobacteriochromes and their Application in the Photoregulation of cAMP Synthesis.

Cyanobacteriochromes (CBCRs) are photoreceptors that bind to a linear tetrapyrrole within a conserved cGMP-phosphodiesterase/adenylate cyclase/FhlA (GAF) domain and exhibit reversible photoconversion. Red/green-type CBCR GAF domains that photoconvert between red- (Pr) and green-absorbing (Pg) forms occur widely in various cyanobacteria. A putative phototaxis regulator, AnPixJ, contains multiple red/green-type CBCR GAF domains. We previously reported that AnPixJ's second domain (AnPixJg2) but not its fourth domain (AnPixJg4) shows red/green reversible photoconversion. Herein, we found that AnPixJg4 showed Pr-to-Pg photoconversion and rapid Pg-to-Pr dark reversion, whereas AnPixJg2 showed a barely detectable dark reversion. Site-directed mutagenesis revealed the involvement of six residues in Pg stability. Replacement at the Leu294/Ile660 positions of AnPixJg2/AnPixJg4 showed the highest influence on dark reversion kinetics. AnPixJg2_DR6, wherein the six residues of AnPixJg2 were entirely replaced with those of AnPixJg4, showed a 300-fold faster dark reversion than that of the wild type. We constructed chimeric proteins by fusing the GAF domains with adenylate cyclase catalytic regions, such as AnPixJg2-AC, AnPixJg4-AC and AnPixJg2_DR6-AC. We detected successful enzymatic activation under red light for both AnPixJg2-AC and AnPixJg2_DR6-AC, and repression under green light for AnPixJg2-AC and under dark incubation for AnPixJg2_DR6-AC. These results provide platforms to develop cAMP synthetic optogenetic tools.

FACING THE EVITABLE AND THE INEVITABLE: PERSPECTIVES OF GOOD DEATH AMONGST MUSLIM PATIENTS AND HEALTH CARE PROVIDERS IN PAKISTAN

Objective: To review the validity of the future of health and care of older people (TFHCOP), good death perception criteria in Muslim patients and health care providers in cultural background of Pakistan.Study Design: A mixed method design with qualitative and quantitative components.Place and Duration of Study: Oncology Department, Combined Military Hospital, Rawalpindi, from July 2015 to October 2015.Material and Methods: It is a sequential explanatory type of mixed method research. A total of 110 participants were included in the study by non probability convenient sampling technique. A modified questionnaire was used based on the principles of TFHCOP good death definition comprising of 8 questions. This was followed by interviews. For data collection and analysis grounded theory approach was used with constant comparisons and open coding. Descriptive statistics were used to analyze questionnaire responses. For bivariate analysis we used chi-square test.Results: There was a consensus on the principles presented in the modified questionnaire in the two groups studied. Total of three domains measured non-significant difference in patient’s and health care provider’s perspectives. Participants identified four more domains. The first domain was related to faith in Allah Almighty, second domain is about leaving the will for bereaved, third domain was about the concerns to finish unfinished agendas and the fourth domain was related to the importance of family security over treatment or death.Conclusion: Perception of good death by Muslim patients and health care providers in Pakistan is different from that of Western and Arab communities in certain domains like religious beliefs, leaving will for the bereaved, finishing unfinished work and the importance of family concerns.

A novel tarantula toxin stabilizes the deactivated voltage sensor of bacterial sodium channel.

Voltage-gated sodium channels (NaVs) are activated by transiting the voltage sensor from the deactivated to the activated state. The crystal structures of several bacterial NaVs have captured the voltage sensor module (VSM) in an activated state, but structure of the deactivated voltage sensor remains elusive. In this study, we sought to identify peptide toxins stabilizing the deactivated VSM of bacterial NaVs. We screened fractions from several venoms and characterized a cystine knot toxin called JZTx-27 from the venom of tarantula Chilobrachys jingzhao as a high-affinity antagonist of the prokaryotic NaVs NsVBa (nonselective voltage-gated Bacillus alcalophilus) and NaChBac (bacterial sodium channel from Bacillus halodurans) (IC50 = 112 nM and 30 nM, respectively). JZTx-27 was more efficacious at weaker depolarizing voltages and significantly slowed the activation but accelerated the deactivation of NsVBa, whereas the local anesthetic drug lidocaine was shown to antagonize NsVBa without affecting channel gating. Mutation analysis confirmed that JZTx-27 bound to S3-4 linker of NsVBa, with F98 being the critical residue in determining toxin affinity. All electrophysiological data and in silico analysis suggested that JZTx-27 trapped VSM of NsVBa in one of the deactivated states. In mammalian NaVs, JZTx-27 preferably inhibited the inactivation of NaV1.5 by targeting the fourth transmembrane domain. To our knowledge, this is the first report of peptide antagonist for prokaryotic NaVs. More important, we proposed that JZTx-27 stabilized the NsVBa VSM in the deactivated state and may be used as a probe to determine the structure of the deactivated VSM of NaVs.-Tang, C., Zhou, X., Nguyen, P. T., Zhang, Y., Hu, Z., Zhang, C., Yarov-Yarovoy, V., DeCaen, P. G., Liang, S., Liu, Z. A novel tarantula toxin stabilizes the deactivated voltage sensor of bacterial sodium channel.

한국 초등학생들을 위한 영어 초기 문해력 측정 도구 모형 개발 연구

The Dynamic Indicators of Basic Early Literacy Skills (DIBELS) are a set of procedures for assessing the acquisition of early literacy skills of native speakers of English. They have been widely perceived as effective in identifying English learners at risk of later reading difficulties and providing them with earlier effective support. It has been argued, however, that an English early literacy test should reflect the situation of English learning in a local educational context. Against this backdrop, this study aims to develop a model of an English early literacy test that could better reflect the characteristics of Korean elementary school students. For this purpose, a comprehensive review was conducted of (a) related research, (b) the current English curriculum for Korean elementary schools, and (c) multiple voices and perspectives from seven experts in English education and twenty elementary school teachers. The model is divided into four domains: alphabetic principle, phonemic awareness, vocabulary, and fluency and comprehension. The first domain is further divided into three tasks, namely reading alphabets, identifying capital letters, and reading pseudowords. The second and third domains are concerned with finding words with the same onsets or rhymes and identifying the meanings of basic English words, respectively. The fourth domain is comprised of two-step tasks measuring fluency and comprehension at both sentence and text levels. This study also offers suggestions about making the model more field-friendly.

Patient-reported outcomes - an approach and application in amniotic membrane transplantation

Abstract Patient-reported outcomes (PROs) is a comparatively novel concept in medicine and in public health services evaluation. It relies on the fact that medical technologies are not able to give all the information about the treatment or a medical condition. Some essential data can be obtained only from the patient. Herewith PROs approach is presented in relation to amniotic membrane transplantation (AMT) which is applied in 306 patients within a period of 7 years who have been treated at Varna Specialized Eye Hospital for different severe anterior ocular surface diseases (AOSD). The patients are divided in seven cohorts according to their diagnosis. The conditions are associated with severe pain, photophobia, secretion and hyperemia. The aim of the study is to present the PROs-approach and the created especially for the study Patient-Reported Outcome Measures (PROMs) tool. Further, to summarize, analyze and discuss the PROs-data of the study. Materials and Methods: The presented PROs-study enrolls 306 hospitalized patients diagnosed in seven cohorts and suitable for AMT for a period of seven years (2010-2016). It is a part of a wider research project. Demographic data and disease-related characteristics were collected in addition to the PROMs questionnaire. The PROMs instrument consists of 4 domains/questions, and the possible answers are put in an ordinal scales. The first domain is Pain , and the possible answers are 5 (0- none; 1- sometimes low; 2- always low; 3- moderate to severe; 4- always severe). The second domain is Photophobia , and the possible answers are 4 (0- none; 1- only at strong light; 2- at daylight; 3- at moderate light). The third domain is Secretion and the possible answers are 4 (0- none; 1- watery; 2- mucus secretion; 3- purulent discharge). The fourth domain is Hyperemia and the possible five answers following the Efron grading scale are (grade 0- normal; grade 1- trace; grade 2- mild; grade 3- moderate; grade 4- severe). Results and Discussion: The AM treatment resulted in significant reduction of the symptoms in all domains (p< 0,001). Regarding Pain domain, in all the groups over 60% of the patients reported improvement. More than 90% of the patients with most severe conditions like keratitis reported improvement. The reported and analyzed results indicate that AMT has a positive impact on all the PRO-domains – Pain, Photophobia, Secretion and Hyperemia, for every patient (p<0.001). This is a novel study for Bulgaria, which aims to measure the impact of AMT on PROs among a large number of patients with severe disorders of AOS divided in seven cohorts for seven years. AMT proved to be a surgical method for treatment of chronic, recurrent anterior ocular surface disease that improves patients’ subjective well-being.

PENGUKURAN TINGKAT KEMATANGAN TATA KELOLA TI BERDASARKAN 34 KERANGKA KERJA COBIT 4.1

Information Technology Support in business is very big because affects the continuity of operational processes of organizations. In its management, Information Technology needs a standard that can help managers to see the gaps between business risks, control needs, and problems. In its management, Information Technology needs a standard that can help managers to see the gaps between business risks, control needs, and problems of existing techniques. IT Governance is a structure of relationships and processes that drive and manage the organization in achieving its objectives by providing value-added of Informatiom Technology utilization and balancing risk with the results provided by Information Technology and processes. Maharaja Ban is a company that is committed to developing business and serving the needs of customers, so it needs the support of good IT Governance. COBIT (Control Objectives for Information and Related Technology) is one tool in the measurement of IT Governance that is used to view existing IT governance. COBIT has 4 domains, namely Plan and Organization (PO), Acquisition and Implementation (AI), Delivery and Support (DS), and Monitoring and Evaluate (ME). This research was conducted for the fourth domain is the restriction on the 34 process . Research results found that the level of maturity (maturity level) IT Governance at Maharaja Ban on level 2. This means that the current maturity level was below the expected level of maturity, so it needs to be improved to be located at the expected level.

Survey, Culture, and Genome Analysis of Ocular Chlamydia trachomatis in Tibetan Boarding Primary Schools in Qinghai Province, China.

Trachoma, the leading infectious cause of blindness worldwide, is an ancient human disease. Its existence in China can be traced back to as early as the twenty-seventh century BC. In modern China, the overall prevalence of trachoma has dramatically reduced, but trachoma is still endemic in many areas of the country. Here, we report that 26 (8%) of 322 students from two rural boarding schools of Qinghai province, west China, were identified as having ocular C. trachomatis infection; and 15 ocular C. trachomatis strains were isolated from these trachoma patients. Chlamydiae in 37 clinical samples were genotyped as type B based on ompA gene analyses. Three ompA variants with one or two in-between SNP differences in the second or fourth variable domain were found. C. trachomatis strains QH111L and QH111R were from the same patient's left and right conjunctival swabs, respectively, but their ompA genes have a non-synonymous base difference in the second variable domain. Moreover, this SNP only exists in this single sample, suggesting QH111L is a newly emerged ompA variant. Interestingly, chromosomal phylogeny analysis found QH111L clusters between a branch of two type B strains and a branch of both A and C strains, but is significantly divergent from both branches. Comparative chromosome analysis found that compared to sequences of reference B/TZ1A828/OT strain, 12 of 22 QH111L's chromosomal genes exhibiting more than nine SNPs have the best homology with reciprocal genes of UGT strains while 9 of 22 genes are closest to those of type C strains. Consistent with findings of UGT-type genetic features in the chromosome, the QH111L plasmid appears to be intermediate between UGT and classical ocular plasmids due to the existence of UGT-type SNPs in the QH111L plasmid. Moreover, the QH111L strain has a unique evolutionarily older cytotoxin region compared to cytotoxin regions of other C. trachomatis strains. The genome analyses suggest that the QH111L strain is derived from recombinations between UGT and classical ocular ancestors. This is the first study of culture and characterization of ocular C. trachomatis in Qinghai Tibetan areas.

Plasma Membrane Origin of the Steroidogenic Pool of Cholesterol Used in Hormone-induced Acute Steroid Formation in Leydig Cells

Hormone-sensitive acute steroid biosynthesis requires trafficking of cholesterol from intracellular sources to the inner mitochondrial membrane. The precise location of the intracellular cholesterol and its transport mechanism are uncertain. Perfringolysin O, produced by Clostridium perfringens, binds cholesterol. Its fourth domain (D4) retains cholesterol-binding properties but not cytotoxicity. We transfected steroidogenic MA-10 cells of mouse Leydig cell tumors with the mCherry-D4 plasmid. Tagged D4 with fluorescent proteins enabled us to track cholesterol. The staining was primarily localized to the inner leaflet of the plasma membrane and was partially released upon treatment with dibutyryl-cAMP (Bt2cAMP), a cAMP analog. Inhibitors of cholesterol import into mitochondria blocked steroidogenesis and prevented release of D4 (and presumably cholesterol) from the plasma membrane. We conclude that the bulk of the steroidogenic pool of cholesterol, mobilized by Bt2cAMP for acute steroidogenesis, originates from the plasma membrane. Treatment of the cells with steroid metabolites, 22(R)-hydroxycholesterol and pregnenolone, also reduced D4 release from the plasma membrane, perhaps evidence for a feedback effect of elevated steroid formation on cholesterol release. Interestingly, D4 staining was localized to endosomes during Bt2cAMP stimulation suggesting that these organelles are on the route of cholesterol trafficking from the plasma membrane to mitochondria. Finally, D4 was expressed in primary rat Leydig cells with a lentivirus and was released from the plasma membrane following Bt2cAMP treatment. We conclude that the plasma membrane is the source of cholesterol for steroidogenesis in these cells as well as in MA-10 cells.

Analysis of the interaction of calcitriol with the disulfide isomerase ERp57.

Calcitriol, the active form of vitamin D3, can regulate the gene expression through the binding to the nuclear receptor VDR, but it can also display nongenomic actions, acting through a membrane-associated receptor, which has been discovered as the disulfide isomerase ERp57. The aim of our research is to identify the binding sites for calcitriol in ERp57 and to analyze their interaction. We first studied the interaction through bioinformatics and fluorimetric analyses. Subsequently, we focused on two protein mutants containing the predicted interaction domains with calcitriol: abb’-ERp57, containing the first three domains, and a’-ERp57, the fourth domain only. To consolidate the achievements we used the calorimetric approach to the whole protein and its mutants. Our results allow us to hypothesize that the interaction with the a’ domain contributes to a greater extent than the other potential binding sites to the dissociation constant, calculated as a Kd of about 10−9 M.

Evaluation of the student learning experience.

This article, the final in a series of 11, discusses the importance of evaluating the students' learning experience. Mentoring is integral to effective nursing care and professional development. It is important that mentors are able to establish whether they are effective in their role, including identifying their strengths and areas for improvement, and whether they are making a difference to the students' learning and contributing to the learning experience and assessment process. The article relates to the fourth domain and outcomes of the Nursing and Midwifery Council's Standards to Support Learning and Assessment in Practice on evaluation of learning and presents several ways in which mentors and practice teachers can evaluate their role and the students' learning experience.

Robust Therapeutic Efficacy of Matrix Metalloproteinase-2-Cleavable Fas-1-RGD Peptide Complex in Chronic Inflammatory Arthritis.

ObjectiveTherapeutic agents that are transformable via introducing cleavable linkage by locally enriched MMP-2 within inflamed synovium would enhance therapeutic efficacy on chronic inflammatory arthritis. Transforming growth factor-β-inducible gene-h3 (βig-h3), which consists of four fas-1 domains and an Arg-Gly-Asp (RGD) motif, intensifies inflammatory processes by facilitating adhesion and migration of fibroblast-like synoviocyte in the pathogenesis of rheumatoid arthritis (RA). The aim of this study was to investigate whether a MMP-2-cleavable peptide complex consisting of a fas-1 domain and an RGD peptide blocks the interaction between βig-h3 and resident cells and leads to the amelioration of inflammatory arthritis.MethodsWe designed βig-h3-derivatives, including the fourth fas-1 domain truncated for H1 and H2 sequences of mouse (MFK00) and MMP-2-cleavable peptide complex (MFK902). MMP-2 selectivity was examined by treatment with a series of proteases. MFK902 efficacy was determined by the adhesion and migration assay with NIH3T3 cells in vitro and collagen-induced arthritis (CIA) model using male DBA/1J mice in vivo. The mice were treated intraperitoneally with MFK902 at different dosages.ResultsMFK902 was specifically cleaved by active MMP-2 in a concentration-dependent manner, and βig-h3-mediated adhesion and migration were more effectively inhibited by MFK902, compared with RGD or MFK00 peptides. The arthritis activity of murine CIA, measured by clinical arthritis index and incidence of arthritic paws, was significantly ameliorated after treatment with all dosages of MFK902 (1, 10, and 30 mg/kg). MFK902 ameliorated histopathologic deterioration and reduced the expression of inflammatory mediators simultaneously with improvement of clinical features. In addition, a favorable safety profile of MFK902 was demonstrated in vivo.ConclusionThe present study revealed that MMP-2-cleavable peptide complex based on βig-h3 structure is a potent and safe therapeutic agent for chronic inflammatory arthritis, thus providing reliable evidence for a MMP-2-cleavable mechanism as a tissue-targeted strategy for treatment of RA.

Erratum to: Molecular characteristics of the ompA gene of serotype B Chlamydia trachomatis in Qinghai Tibetan primary school students.

To study the molecular characteristics of Chlamydia trachomatis, the major outer membrane protein gene (ompA) of C. trachomatis from primary school students with trachoma residing in the Qinghai Tibetan area was sequenced and compared with the same serotype in GenBank. In Jianshetang Primary School and Galeng Central Primary School in the Galeng Tibetan Township of Qinghai Haidong Sala Autonomous County, scraped samples were collected from the upper tarsal conjunctiva and lower conjunctival sac of both eyes of 45 students with trachoma, stored at 4°C, and transported to Beijing Tongren Hospital by air within 24 h. The samples were screened for C. trachomatis by real-time PCR. The ompA gene from the C. trachomatis-positive samples was amplified by nested PCR. The serotype was confirmed by National Center for Biotechnology Information (NCBI) BLAST search and homology analysis. The entire ompA gene sequence was compared with the corresponding gene sequences of serotype B strains available in GenBank. Of the 45 students aged 6–13 years with trachoma, 26 C. trachomatis-positive students were identified by the initial real-time PCR screening (average age, (9.09±1.63) years; sex ratio, 1.0), accounting for 57.78% (26/45). The cycle threshold values for real-time PCR were 16.79–37.77. Half (13/26) of C. trachomatis-positive students had a bacterial copy number of >105. The compliance rate of the ompA gene sequences with the C. trachomatis serotype B strains in GenBank was up to 99%. Two novel genetic mutations were found when the ompA gene was compared with those of the 11 serotype B strains in GenBank. The two non-synonymous mutations were located at (i) position 271 in the second constant domain, an adenine (A) to guanine (G) substitution (ACT→GCT), changing the amino acid at position 91 from threonine to alanine (Thr→Ala) in all 26 strains; and (ii) position 887 in the fourth variable domain, a cytosine (C) to thymine (T) substitution (GCA→GTA), changing the amino acid at residue 296 from alanine to valine (Ala→Val) in four of the 26 strains. Six mutations were identified relative to ATCC VR-573. The strains could be divided into two gene clusters according to the mutation at nucleotide position 887: CQZ-1 (China Qinghai Tibetan-1) and CQZ-2 (China Qinghai Tibetan-2). We thus detected two novel serotype B mutant strains of C. trachomatis among study subjects with trachoma.

The Role of the Liberal Arts in Preparing Health Education Specialists

Purpose. Exposure to a liberal education or a set of liberal courses or curriculum is an approach to college learning that empowers individuals and prepares them to deal with complexity, diversity, and change in a competitive global economy. Public health education is a diverse, ever-changing field composed of complex challenges. Undergraduate student preparation for entry-level public health education specialists requires the mastery of the National Commission for Health Education Credentialing’s Health Education Specialist’s seven Areas of Responsibility. Due to the overlap in the Health Education Specialist Practice Analysis (HESPA) Areas of Responsibilities and the Liberal Arts (LA) guiding principles (three pillars) and Liberal Education and America’s Promise (LEAP) learning essential outcomes, liberal arts universities are uniquely positioned to offer undergraduate public health education programs. Method. Health education specialist competencies and subcompetencies were mapped onto liberal arts pillars and LEAP learning essential outcomes. Public health education course activities from two public liberal arts universities were used to demonstrate the application of health education competencies between the LA pillars and LEAP learning essential outcomes. Results. LEAP essential learning outcome Domains 1 through 3 provide a foundation for and the development of HESPA Areas I, IV, VI, and VII. A fourth domain addresses Areas III and V. Conclusion. Liberal arts universities are uniquely positioned to provide students with an undergraduate degree in the increasingly popular public health education major due to the established parallels between liberal education and health education specialist learning outcomes.

Evaluation of Madrasah Education in Elementary Schools of Digos City

This study on evaluation of Madrasah Education in elementary schools of Digos City was conducted to determine the demographic profile of Asatidz, the problems met in their respective Madrasah and their curriculum implementation. To measure the Asatidz competency level in teaching Arabic Language and Islamic Values, the tool used was taken from Curriculum Management Assessment Data culled from the fourth domain of the Philippine National Competency-Based Teacher Standard (NCBTS). The Descriptive-correlational method was used in this study. The results of responses coming from all Asatidz in nine (9) Madaris of Digos City were used in the analysis using the relative frequency, mean, standard deviation, and Pearson r. Results of the study disclosed that there was no significant relationship between the demographic profile of the Asatidz and the extent of their Madrasah curriculum implementation. Also, on the extent of the problems met by the Asatidz and the extent of their Madrasah curriculum implementation did not show a significant relationship. However on educational attainment, there was a highly significant relationship in the implementation of Madrasah curriculum. It is recommended that the DepEd must design policies that will provide more education trainings for Asatidz to become more capable of helping students to acquire the competencies needed to evolve in today‘s society.

En domänteori för organisering av socialt arbete i offentlig sektor

A domain theory for the organization of social work in the public sector The article presents outlines of a domain theory for the organization of social work in the public sector. e aim is to describe – from a Nordic perspective – how social workers’ professional prac- tice is shaped by local level politics and administration, and how such shaping processes at the local level are conditioned and controlled by general structural and institutional conditions at the national and international level. We argue that public sector arenas for social work are constituted by three domains with different tasks and logics, yet interconnected and interdependent: the policy domain, the domain of administration and the profession’s domain. Organizations at the local level – constituted by the three domains and their respective logics – are, in turn, conditioned by a fourth domain: the domain of institutional conditions. The latter domain is about control ideals, norms, knowledge, epistemological approaches and technologies prevailing in society, legitimized and disseminated by central institutions that affect how local level organizations operate.

Crystal structures of the ligand-binding region of uPARAP: effect of calcium ion binding.

The proteins of the mannose receptor (MR) family share a common domain organization and have a broad range of biological functions. Urokinase plasminogen activator receptor-associated protein (uPARAP) (or Endo180) is a member of this family and plays an important role in extracellular matrix remodelling through interaction with its ligands, including collagens and urokinase plasminogen activator receptor (uPAR). We report the crystal structures of the first four domains of uPARAP (also named the ligand-binding region, LBR) at pH7.4 in Ca(2+)-bound and Ca(2+)-free forms. The first domain (cysteine-rich or CysR domain) folds into a new and unique conformation different from the β-trefoil fold of typical CysR domains. The so-called long loop regions (LLRs) of the C-type lectin-like domain (CTLD) 1 and 2 (the third and fourth domain) mediate the direct contacts between these domains. These LLRs undergo a Ca(2+)-dependent conformational change, and this is likely to be the key structural determinant affecting the overall conformation of uPARAP. Our results provide a molecular mechanism to support the structural flexibility of uPARAP, and shed light on the structural flexibility of other members of the MR family.

(1)H, (15)N, and (13)C resonance assignments of Staphylococcus aureus extracellular adherence protein domain 4.

The pathogenic bacterium Staphylococcus aureus has evolved to actively evade many aspects of the human innate immune system by expressing a series of secreted inhibitory proteins. Among these, the extracellular adherence protein (Eap) has been shown to inhibit the classical and lectin pathways of the complement system. By binding to complement component C4b, Eap is able to inhibit formation of the CP/LP C3 pro-convertase. Secreted full-length, mature Eap consists of four ~98 residue domains, all of which adopt a similar beta-grasp fold, and are connected through a short linker region. Through multiple biochemical approaches, it has been determined that the third and fourth domains of Eap are responsible for C4b binding. Here we report the backbone and side-chain resonance assignments of the 11.3kDa fourth domain of Eap. The assignment data has been deposited in the BMRB database under the accession number 26726.

Molecular characteristics of the ompA gene of serotype B Chlamydia trachomatis in Qinghai Tibetan primary school students.

To study the molecular characteristics of Chlamydia trachomatis, the major outer membrane protein gene (ompA) of C. trachomatis from primary school students with trachoma residing in the Qinghai Tibetan area was sequenced and compared with the same serotype in GenBank. In Jianshetang Primary School and Galeng Central Primary School in the Galeng Tibetan Township of Qinghai Haidong Sala Autonomous County, scraped samples were collected from the upper tarsal conjunctiva and lower conjunctival sac of both eyes of 45 students with trachoma, stored at 4°C, and transported to Beijing Tongren Hospital by air within 24 h. The samples were screened for C. trachomatis by real-time PCR. The ompA gene from the C. trachomatis-positive samples was amplified by nested PCR. The serotype was confirmed by National Center for Biotechnology Information (NCBI) BLAST search and homology analysis. The entire ompA gene sequence was compared with the corresponding gene sequences of serotype B strains available in GenBank. Of the 45 students aged 6-13 years with trachoma, 26 C. trachomatis-positive students were identified by the initial real-time PCR screening (average age, (9.09±1.63) years; sex ratio, 1.0), accounting for 57.78% (26/45). The cycle threshold values for real-time PCR were 16.79-37.77. Half (13/26) of C. trachomatis-positive students had a bacterial copy number of >10(5). The compliance rate of the ompA gene sequences with the C. trachomatis serotype B strains in GenBank was up to 99%. Two novel genetic mutations were found when the ompA gene was compared with those of the 11 serotype B strains in GenBank. The two non-synonymous mutations were located at (i) position 271 in the second constant domain, an adenine (A) to guanine (G) substitution (ACT→GCT), changing the amino acid at position 91 from threonine to alanine (Thr→Ala) in all 26 strains; and (ii) position 887 in the fourth variable domain, a cytosine (C) to thymine (T) substitution (GCA→GTA), changing the amino acid at residue 296 from alanine to valine (Ala→Val) in four of the 26 strains. Six mutations were identified relative to ATCC VR-573. The strains could be divided into two gene clusters according to the mutation at nucleotide position 887: CQZ-1 (China Qinghai Tibetan-1) and CQZ-2 (China Qinghai Tibetan-2). We thus detected two novel serotype B mutant strains of C. trachomatis among study subjects with trachoma.

Numerical Simulations of Typhoon Morakot (2009) Using a Multiply Nested Tropical Cyclone Prediction Model

Abstract High-impact Typhoon Morakot (2009) was investigated using a multiply nested regional tropical cyclone prediction model. In the numerical simulations, the horizontal grid spacing, cumulus parameterizations, and microphysical parameterizations were varied, and the sensitivity of the track, intensity, and quantitative precipitation forecasts (QPFs) was examined. With regard to horizontal grid spacing, it is found that convective-permitting (5 km) resolution is necessary for a reasonably accurate QPF, while little benefit is gained through the use of a fourth domain at 1.67-km horizontal resolution. Significant sensitivity of the track forecast was found to the cumulus parameterization, which impacted the model QPFs. In particular, the simplified Arakawa–Schubert parameterization tended to erroneously regenerate the remnants of Tropical Storm Goni to the southwest of Morakot, affecting the large-scale steering flow and the track of Morakot. Strong sensitivity of the QPFs to the microphysical parameterization was found, with the track and intensity showing little sensitivity. It is also found that Morakot’s accumulated precipitation was reasonably predictable, with the control simulation producing an equitable threat score of 0.56 for the 3-day accumulated precipitation using a threshold of 500 mm. This high predictability of precipitation is due in part to more predictable large-scale and topographic forcing.

Androgen-inducible gene 1 increases the ER Ca2 + content and cell death susceptibility against oxidative stress

Androgen-induced gene 1 (AIG1) is a transmembrane protein implicated with survival (its expression level was shown to correlate with the survival of patients suffering from hepatocellular carcinoma) and Ca2+ signaling (over-expression of AIG1 increased transcription mediated by the Ca2+-dependent nuclear factor of activated T cells). We aimed to shed light on this less-studied protein and investigated its tissue expression, genomic organization, intracellular localization and membrane topology as well as its effects on cell death susceptibility and the Ca2+ content of the endoplasmic reticulum. Immunoblotting of mouse tissues demonstrated highest expression of AIG1 in the liver, lung and heart. AIG1 has a complex genomic organization and expresses several splice variants in a tissue-dependent manner. Analyzing the topology of AIG1 in the ER membrane using a protease-protection assay suggested that AIG has five transmembrane domains with a luminal N- and cytosolic C-terminus and a hydrophobic stretch between the third and fourth membrane domain that does not cross the membrane. AIG1 over-expression slightly increased susceptibility to oxidative stress, which correlated with an increased ER Ca2+ concentration in two different cell lines. Together, these results indicate that AIG1 plays a role in the control of the intracellular Ca2+ concentration and cell death susceptibility.