bluuebunny/crossref_metadata_2025 huggingface dataset:
Mobile Floriculture Operators in Southern Sri Lanka: Challenges and Solutions
Authors: Anuruddi H. I. G. K. , Fonseka D. L. C. K. , Wickramaarachchi W. W. U. I.
Published: July 2020

Floriculture is a flourishing industry in Sri Lanka with a great potential in both domestic and export market. Street and mobile flower vending is a micro enterprise which is not a negligible portion of floriculture sector in the country. Present study was conducted to investigate the constraints faced by street and mobile flower vendors and to understand their willingness to adopt those challenges. Randomly selected 40 flower vendors from Galle, Matara and Hambantota districts were interviewed using a semi structured questionnaire by visiting them at their marketing sites such as streets, week fairs and “Kaprukawaruna” exhibitions conducted by Coconut Cultivation Board, Matara. According to the findings, majority of the respondents engage in the business as a full time employment and 43.3% of the vendors sell plants grown in their own nurseries. Apart from the flowering and ornamental plants, they sell fertilizers, coir dust and coconut husk pieces and product diversification has a significant impact (p=0.016) on their monthly income. Average nursery size was found to be 500ft2 where average income was noted as 200 USD. Among the vendors 70% use their own money to refinance the business. Majority of the respondents (60%) were members of different flower grower societies. Vendors (87.5%) sell their products targeting week fairs and (62.5%) sell their products beside the streets in temporary fixed huts. Not having an authorized place to market their goods was identified as the major problem by 85% of the respondents. Majority of the vendors (62.5%) were willing to expand the business while 42.5% identified this as a profitable venture to invest. According to the binary logistic regression results, age (p=0.000), education level (p=0.001), economic status (p=0.057), nursery size (p=0.004) and credit accessibility (p=0.024) were positively related with their willingness to adopt for the challenges they face. It is concluded that following key actions must be taken to strengthen the street and mobile flower vendors for their further empowerment: Provision of proper authorized sites of operation, improve the market credit and other infrastructure facilities and more training opportunities.
Evaluation of Short Duration Drought Tolerant Rice Varieties in Drought Prone Areas of Subarnapur District of Odisha
Authors: Pradhan S. , Rahman F. H. , Sethy S. , Pradhan G. , Sen J.
Published: July 2020

A study was conducted in farmer’s field through technology demonstrations during Kharif season of 2018 and 2019 in Subarnapur district of Odisha to recommend the most suited short duration drought tolerant upland rice variety particular to the locality. Six different varieties like Sahabhagi dhan, Mandakini, Jogesh, DRR-44, and DRR-42 & Khandagiri were taken with four replications in Randomized Block Design in both years. Technology demonstration included not only the improved short duration varieties, but also a package of good agronomic practices viz. line sowing, seed treatment with Bavistin, proper fertilization with dose @ 80:40:40 NPK/ha, pre emergence herbicide application with pretilachlor, needful manual weeding and  need based plant protection chemicals. The results of demonstration showed that farmers could increase rice productivity notably by adopting improved variety Sahabhagi dhan and DRR-44. These varieties recorded higher grain yields of 64% and 46%, respectively, and fetched a sum return of Rs 25620 and Rs 25275 over farmers’ local variety Khandagiri.
Molecular Characterization and Detection of Antibiotic Resistance Genes in Pseudomonas Species Isolated from Tympanotonus fuscatus
Authors: Sampson T. , Akani N. P. , Hakam I. O.
Published: July 2020

Aim: This was carried out to characterize Pseudomonas species isolated from the West African Mud Creeper (Tympanotonus fuscatus) molecularly and as well detect the possible presence of inducible AmpC gene that mediates resistance to cephalosporins and most penicillins.

Sample: Tympanotonus fuscatus (West African Mud Creeper), a gastropod mollusc found in brackish waters of West Africa was used for the study.

Place and Duration of Study: This study was carried out between February and August 2019 at the Department of Microbiology, Rivers State University, Port Harcourt, Nigeria.

Methodology: Thirty two (32) Pseudomonas species were isolated and identified culturally from T. fuscatus. Pseudomonas species isolates were subjected to a group of ten (10) antibiotics using the Kirby-Bauer disc diffusion method and resistant isolates were screened molecularly for the presence of resistance gene (AmpC). AmpC screening was carried out in a step wise process of DNA extraction, quantification, amplification of ampC gene using appropriate primer and Agarose gel electrophoresis to reveal which DNA extracts had ampC genes amplified. The two most resistant isolates had their 16S rRNA sequenced, identified and were also profiled for plasmids by extracting plasmid DNA.

Results: Results revealed that 96.67% of isolates had MAR index greater than 0.2 indicating high a risk source of contamination where antibiotics are often used. Results also showed the presence of ampC gene in seven (7) out of the twelve (12) isolates screened for ampC gene. Molecular characterization via sequencing of the 16S rRNA gene of the two (2) most resistant isolates confirmed that both isolates were strains of Pseudomonas aeruginosa. Profiling of plasmids also revealed the presence of plasmid DNA of about 10 kilo base pairs in both isolates profiled.

Conclusion: This study has revealed the resistance ability of Pseudomonas and some reasons behind this resistance. Appropriate investigation into antimicrobial resistance is recommended for the administration of drugs for the treatment of food-mediated Pseudomonas infections.
Prevalent Beliefs and Attitude toward Vitiligo among Public in Al Baha Province, Southern Saudi Arabia
Authors: Ghamdi Hasan S. AL- , Alzahrani Mohammed Ali M. , Alsohaimi Aziz , Alzahrani Saif Abadi , Alzahrani Mohammed Hassan , AlGhamdi Osama Saeed Ali , AlGhamdi Mohammed Abdullah M.
Published: July 2020

Introduction: Vitiligo is an apparent skin condition that has a psychological impact on the affected individuals and their families. Its prevalence is estimated to be about (0.5-2%) globally. Misunderstandings, and negative attitudes toward vitiligo patients are the most important factors affecting the quality of life of vitiligo patients.

Objective: The objective of this study was to identify the common misconceptions and attitudes about the nature, causes and prognosis of vitiligo in Al-Baha region, Saudi Arabia.

Methods: A cross sectional study collected data from the general population in the public malls in Al-Baha region, Saudi Arabia. We used a questionnaire which was developed by the authors, the questionnaire included questions regarding vitiligo cause, nature, prognosis and the attitude toward vitiligo.

Results: A total of (453) responses were analysed, of which (53%) were females, and (90.9%) were Saudis. The participants’ ages ranged from (16 -64) with a mean of (28.3). The majority of our sample had a high level of education; university (65.1%), High school (26%) and less than high school (6%). Our population believed that gecko and its saliva were the commonest cause (38.1%). Most of the respondents (44.4%) thought that the condition is exaggerated by stress and anxiety. Regarding attitude; (31.5%) are unwilling to marry a vitiligo patient.

Conclusion: The results of the present study show that the misconceptions and attitudes toward vitiligo are prevalent. Health awareness campaigns should be implanted to educate the people and improve the quality of life of the affected individuals.
Molecular Identification of Virulence Genes of Escherichia coli Isolated from Cow Milk and Its Products in Abuja, Nigeria
Authors: Okechukwu E. C. , Amuta E. U. , Gberikon G. M. , Chima N. , Yakubu B. , Igwe J. C. , Njoku M.
Published: July 2020

Shiga toxin-producing Escherichia coli have been identified as an emerging foodborne pathogen which portends serious risk to human health. Cow milk and its products are potential sources of shiga toxin-producing Escherichia coli. A relatively small number from the family of shiga toxin-producing Escherichia coli are pathogenic. It becomes necessary that Cow milk and milk products are regularly screened for the presence of virulence genes in microbes. The study aimed to genetically determine the presence of virulence genes that are characteristic of Enterohaemorrhagic E. coli in 600 milk samples. The E. coli isolates were recovered from the milk samples (n=35), biochemically examined and genetically screened for virulence genes by multiplex Polymerase Chain Reaction (PCR). The results of the molecular profiling revealed that (stx2) was detected in 17(60.7%), (hlyA) 11(39.3%) and eae genes 8(28.6%) of the E. coli isolates respectively, while (stx1) was not detected. The results indicated a high prevalence of virulent shiga toxin-producing Escherichia coli in the milk samples. Priority attention should be given to this microbe as it will demand stringent steps in the detection given that they are known to be rigorous in identification.
Evidence for a conformational change in deglycosylated glycoprotein hormones
Authors: Keutmann Henry T. , Johnson Leslie , Ryan Robert J.
Published: July 2002

Carbohydrate
Glycoprotein
Follitropin
Choriogonadotropin
Conformation
Membrane receptor
Mucin‐carbohydrate directed monoclonal antibody
Authors: Kurosaka Akira , Fukui Shigeyuki , Kitagawa Hiroshi , Nakada Hiroshi , Numata Yoshito , Funakoshi Ikuo , Kawasaki Toshisuke , Yamashina Ikuo
Published: July 2002

To raise monoclonal antibodies recognizing cancer‐associated alterations of the carbohydrate structure of glycoproteins, Balb/c mice were immunized with human colonic cancer cells (LS 180 from ATCC). One of the generated hybridomas produced a monoclonal antibody that bound to the carbohydrate moiety of mucin‐type glycoproteins from LS 180. The antibody did not bind to glycoproteins from another colonic cancer cell line, SW 1116, or to glycolipids from any of the colonic cancer cell lines. The antibody bound to ovine and bovine submaxillary mucins (OSM and BSM). NeuAcα2→6Ga1NAc seemed to be involved in the epitope.
Binding of C‐reactive protein to human neutrophils
Authors: Buchta Richard , Pontet Michel , Fridkin Mati
Published: July 2002

The binding of radiolabelled 125I‐CRP to human neutrophils has been characterised according to pH, temperature and time dependence. The binding of 125I‐CRP was saturable, very fast (<2 min at 22°C), and the labelled protein was displaced by unlabelled CRP and aggregated human IgG. The dissociation constant was 3.2 × 10−8 M at pH 7.4, 22°C and 8.8 × 10−8 M at pH 6.0, 22°C. The calculated number of binding sites was 5–20 × 104 per cell at pH 7.4, 22°C. An association with an Fc‐type receptor is suggested, since aggregated IgG was able to displace specifically CRP.
Mitochondrial ribosomes of yeast: Isolation of individual proteins and N‐terminal sequencing
Authors: Graack Hanns-Rüdiger , Grohmann Lutz , Choli Theodora
Published: July 2002

Proteins of the small and large subunits of mitochondrial ribosomes from the yeast Saccharomyces cerevisiae were isolated and characterized by two‐dimensional gel electrophoresis. Ribosomal proteins of the large subunit were separated by reverse‐phase HPLC and up to 37 amino acid residues of the N‐terminal sequences of L3, L4, L9 and L31 were determined. No significant homology to ribosomal protein sequences so far determined from other organisms was found.
NMR study of the alkaline isomerization of ferricytochrome c
Authors: Hong Xiaole , Dixon Dabney White
Published: July 2002

The pH‐induced isomerization of horse heart cytochrome c has been studied by 1H NMR. We find that the transition occurring in D2O with a pK
a measured as 9.5±0.1 is from the native species to a mixture of two basic forms which have very similar NMR spectra. The heme methyl peaks of these two forms have been assigned by 2D exchange NMR. The forward rate constant (native to alkaline cytochrome c) has a value of 4.0±0.6 s−1 at 27°C and is independent of pH; the reverse rate constant is pH‐dependent. The activation parameters are ΔH
‡ = 12.8±0.8 kcal·mol1, ΔS
‡ = − 12.9±2.0 e.u. for the forward reaction and ΔH
‡ = 6.0±0.3 kcal·mol−1, ΔS
‡ = − 35.1±1.3 e.u. for the reverse reaction (pH*=9.28). ΔH° and ΔS° for the isomerization are 6.7±0.6 kcal·mol−1 and 21.9±1.0 e.u., respectively.
Characterization of DHP binding protein in crayfish striated muscle
Authors: Krizanova O. , Novotova M. , Zachar J.
Published: July 2002

The dihydropyridine calcium channel blocker, [3H]PN 200‐110, binds specifically also to crayfish muscle membranes, though with a binding capacity smaller than that measured with rabbit or human skeletal muscle membranes. [3H]PN 200‐110 binding proteins from the crayfish T‐tubules were solubilized and purified on WGA Sepharose or extracted from gel. The purified protein has a molecular mass of approximately 190 kDa under nonreducing conditions and was able to transport calcium after reconstitution. Polyclonal antibodies against crayfish T‐tubules enriched with purified DHP‐binding protein were shown to bind to DHP‐binding protein from both the crayfish and the rabbit skeletal muscle, although not with the same intensity. Electron microscopy showed the presence of ovoid particles. Our results suggest that a voltage‐dependent calcium channel may be present in crayfish skeletal muscle, which is homological with the L‐type calcium channel in rabbit skeletal muscle.
Cloning and sequencing of glutamate mutase component S from Clostridium tetanomorphum Homologies with other cobalamin‐dependent enzymes
Authors: Marsh E.Neil G. , Holloway Daniel E.
Published: July 2002

The gene encoding component S, the small subunit1 of glutamate mutase, an adenosylcobalamin (coenzyme B12)‐dependent enzyme from Clostridium tetanomorphum has been cloned and its nucleotide sequence determined. The mutS gene encodes a protein of 137 amino acid residues, with M
r, 14,748. The deduced amino acid sequence showed homology with the C‐terminal portion of adenosylcobalamin‐dependent methylmalonyl‐CoA mutase [1989, Biochem. J. 260, 343–352] and a region of cobalamin‐dependent methionine synthase which has been shown to bind cobalamin [1989, J. Biol. Chem 264, 13888–13895].
Proteolytic processing of human intestinal lactase‐phlorizin hydrolase precursor is not a prerequisite for correct sorting in Madin Darby canine kidney (MDCK) cells
Authors: Grünberg Jürgen , Luginbühl Ursula , Sterchi Erwin E.
Published: July 2002

Maturation or lactase‐phlorizin hydrolase (LPH) (EC 3.2.1.23–62) requires proteolytic processing of precursor (pro‐LPH) to mature microvillus membrane enzyme (m‐LPH). Subcellular site and function of this processing are unknown. We studied the processing and sorting of human LPH expressed permanently in MDCK cells. LPH was inserted into the apical membrane and small amounts were found basolateral. Of the LPH immunoprecipitated from the apical membrane, 42% was in the mature, i.e. proteoytically processed form; on the basolateral membrane it was 20%. Thus, LPH‐processing occurs after sorting and is not necessary for surface expression.
A protein histidine kinase induced m rat liver by peroxisome proliferators
Authors: Motojima Kiyoto , Goto Sataro
Published: July 2002

A novel protein kinase is induced in rat liver plasma membrane by the administration of peroxisome proliferators. A 36 kDa protein (P36) on the membrane was rapidly phosphorylated in vitro by the kinase and the phosphorylated amino acid was identified as phosphohistidine. Histidine phosphorylation of P36 was activated in vitro by recombinant Ras protein and GTP; both decreased Michaelis constant (K
m) for ATP from 1.25 to 0.25 μM. The novel histidine kinase, products of which have been overlooked due to their acid lability, may participate in cellular signaling and peroxisome proliferators may perturb the pathway.
The PPZ protein phosphatases are involved in the maintenance of osmotic stability of yeast cells
Authors: Posas Francesc , Casamayor Antonio , Ariño Joaquín
Published: July 2002

We have recently reported the existence in the yeast Saccharomyces cerevisiae of a gene named PPZ1, encoding a novel Ser/Thr phosphatase characterized by a large, Ser‐rich amino‐terminal extension, and suggested the existence of a related gene product that could have overlapping functions. We have now amplified by polymerase chain reaction techniques a genomic fragment of about 600 bp corresponding to this second gene (PPZ2). This fragment hybridizes to an mRNA of about the same size as the PPZ1 message but the amount of PPZ2 mRNA peaks at the stationary phase, when almost no PPZ1 mRNA is found. The PPZ2 fragment was interrupted in vitro and used to transform diploid heterozygous ppz1
PPZ2 cells. Haploid cells carrying the double mutation ppz1ppz2 were unable to grow in the presence of 5 mM caffeine. However, the mutants did survive when osmotically stabilized in the presence of 1 M sorbitol. The evidence obtained suggests that PPZ1 and PPZ2 may be structurally and functionally related and points to an involvement of these phosphatases in functions related to the maintenance of cell integrity.
Proteolytic processing of the hepatocyte growth factor/scatter factor receptor by furin
Authors: Komada Masayuki , Hatsuzawa Kiyotaka , Shibamoto Sayumi , Ito Fumiaki , Nakayama Kazuhisa , Kitamura Naomi
Published: July 2002

The hepatocyte growth factor/scatter factor (HGF/SF) receptor consists of an α‐ and a β‐subunit, which are derived from a single‐chain precursor by endoproteolytic processing. The precursor is not proteolytically processed in LoVo colon carcinoma cells. The uncleaved receptor immunopurified from the cells was cleaved in vitro by furin. Furthermore, the HGF/SF receptor was proteolytically processed in LoVo cells transfected with furin cDNA. These results indicate that furin is a processing endoprotease for the HGF/SF receptor. Tyrosine autophosphorylation of the uncleaved receptor was induced by HGF/SF, and the growth of the cells expressing the uncleaved receptor was stimulated by HGF/SF, indicating that the proteolytic processing of the receptor is not essential for the signal transduction of HGF/SF.
Cloning and functional expression in Escherichia coli of a cyanobacterial gene for lycopene cyclase, the enzyme that catalyzes the biosynthesis of β‐carotene
Authors: Cunningham Francis X. , Chamovitz Daniel , Misawa Norihiko , Gantt Elisabeth , Hirschberg Joseph
Published: July 2002

Carotenoids with cyclic end groups are essential components of the photosynthetic membrane in all known oxygenic photosynthetic organisms. These yellow pigments serve the vital role of protecting against potentially lethal photo‐oxidative damage. Many of the enzymes and genes of the carotenoid biosynthetic pathway in cyanobacteria, algae and plants remain to be isolated or identified. We have cloned a cyanobacterial gene encoding lycopene cyclase, an enzyme that converts the acyclic carotenoid lycopene to the bicyclic molecule β‐carotene. The gene was identified through the use of an experimental herbicide, 2‐(4‐methylphenoxy)triethylamine hydrochloride (MPTA), that prevents the cyclization of lycopene in plants and cyanobacteria. Chemically‐induced mutants of the cyanobacterium Synechococcus sp. PCC7942 were selected for resistance to MPTA, and a mutation responsible for this resistance was mapped to a genomic DNA region of 200 bp by genetic complementation of the resistance in wild‐type cells. A 1.5 kb genomic DNA fragment containing this MPTA‐resistance mutation was expressed in a lycopene‐accumulating strain of Escherichia coli. The conversion of lycopene to β‐carotene in these cells demonstrated that this fragment encodes the enzyme lycopene cyclase. The results indicate that a single gene product, designated lcy, catalyzes both of the cyclization reactions that are required to produceβ‐carotene from lycopene, and prove that this enzyme is a target site of the herbicide MPTA. The cloned cyanobacterial lcy gene hybridized well with genomic DNA from eukaryotic algae, thus it will enable the identification and cloning of homologous genes for lycopene cyclase in algae and plants.
Selective ubiquitination of calmodulin by UBC4 and a putative ubiquitin protein ligase (E3) from Saccharomyces cerevisiae
Authors: Parag Hadas A. , Dimitrovsky Deborah , Raboy Bilha , Kulka Richard G.
Published: July 2002

A putative ubiquitin protein ligase (E3‐CaM) which cooperates with UBC4 in selectively ubiquitinating calmodulin has been partially purified from Saccharomyces cerevisiae. Ca2+ was required for this activity and monoubiquitinated calmodulin was the main product of the reaction. The apparent K
m of E3‐CaM for calmodulin was approximately 1 μM which is of the same order of magnitude as the concentration of calmodulin in yeast cells. Proteins which are good substrates for other E3s (E3α or E3‐R) were not ubiquitinated by E3‐CaM. Lower but significant activities of E3‐CaM were observed when UBC1 replaced UBC4.
Role of a serum phospholipase A1in the phosphatidylserine‐induced T cell inhibition
Authors: Bellini Fabrizio , Bruni Alessandro
Published: July 2002

We have previously shown that unsaturated phosphatidylserines inhibit mitogen‐induced T cell activation. We now report that the inhibitory action requires a protein present in bovine and human serum. Partial purification and phospholipase assay show that this protein has phospholipase A activity on phosphatidylserine but not phosphatidylethanolamine, phosphatidylcholine and phosphatidylinositol. In short incubations (1–3 h) 2‐acyl lysophosphatidylserine is produced but in longer incubations thecis‐unsaturated fatty acid is also released. Experiments on peripheral blood mononuclear cells indicate that the unsaturated fatty acid becomes the main responsible for the PS‐induced inhibition and that 2‐acyl lysophosphatidylserine enhances the inhibitory effect of fatty acid.
The human serotonin 5‐HT2B receptor: Pharmacological link between 5‐HT2 and 5‐HT1D receptors
Authors: Choi Doo-Sup , Birraux Guillaume , Launay Jean-Marie , Maroteaux Luc
Published: July 2002

The human serotonin 5‐HT2B receptor, isolated from a human liver cDNA library, was transfected in COS‐1 cells. Its pharmacological profile shows divergence with serotonin 5‐HT2B receptors of other species. In particular, although strong correlation is observed between the human and the rat 5‐HT2B receptor pharmacology, the correlation is almost as significant for the mouse 5‐HT2B and the human 5‐HT1D receptor agonists. The major sites of expression of its mRNA are in the human liver and kidney, with detectable expression in lung and heart. Therefore, this human 5‐HT2B receptor could account for functions attributed to the peripheral 5‐HT1D/5‐HT2‐like receptors, especially in the cardiovascular system. Thus, its detailed original pharmacology is of prime importance for therapeutic drug development.
Aequorea green fluorescent protein
Authors: Inouye Satoshi , Tsuji Frederick I.
Published: July 2002

Expression of the cDNA for Aequorea green fluorescent protein in E. coli yielded a fused protein with fluorescence excitation and emission spectra virtually identical to those of the native green fluorescent protein. Further, a solution of the protein, when mixed with aequorin and calcium ion, emitted a greenish luminescence characteristic of the in vivo luminescence of the animal, indicating a radiationless energy transfer to the protein.
Evidence of internucleosomal DNA fragmentation and identification of dying cells in X‐ray‐induced cell death in the developing brain
Authors: Ferrer I. , Macaya A. , Blanco R. , Olivé M. , Cinós C. , Munell F. , Planas A.M.
Published: July 2002

Newborn Sprague‐Dawley rats received a single dose of 2 Gy X‐rays and were killed 6 hr later. Dying cells were characterized by extreme chromatin condensation and nuclear fragmentation. Dying cells were distributed in the primary and secondary germinal zones and in other brain regions. Among these latter, dying cells occurred in the cortical layers of the olfactory bulb, layers II–III and VIb of the neocortex, piriform and entorhinal cortex, stratum oriens and pyramidale of the hippocampus, striatum, thalamus, amygdala, brainstem, internal granular layer of the cerebellum, and cerebral and cerebellar white matter. Dying cells were immature cells, neurons and glial cells (including radial glia). In‐situ labeling of nuclear DNA fragmentation identified individual cells bearing fragmented DNA. Since the number of cells stained with this method was larger than the number of dying cells, as revealed with current histological techniques, it is suggested that nuclear DNA fragmentation precedes chromatin condensation and nuclear fragmentation in X‐ray‐induced apoptosis. Furthermore, agarose gel electrophoresis of extracted DNA from irradiated brains showed a “ladder” pattern which is typical of internucleosomal DNA fragmentation and endonuclease activation.
Molecular identification of a glucose transporter from fish muscle1
Authors: Planas Josep V. , Capilla Encarnación , Gutiérrez Joaquim
Published: July 2002

In mammals and birds, several isoforms of facilitative glucose transporters have been identified (GLUT1–4), but no information is available regarding the molecules involved in glucose transport in other vertebrates. Here we report the cloning of a GLUT molecule from fish muscle with high sequence homology to GLUT4 and containing features characteristic of a functional GLUT. Fish GLUT is expressed predominantly in skeletal muscle, kidney and gill, which are tissues with known high glucose utilization. These results indicate that fish GLUT is structurally, and perhaps functionally, similar to the other known GLUTs expressed in muscle in mammalian and avian species.
Transduction of Cu,Zn‐superoxide dismutase mediated by an HIV‐1 Tat protein basic domain into mammalian cells
Authors: Kwon Hyeok Yil , Eum Won Sik , Jang Hyun Woo , Kang Jung Hoon , Ryu Jiyoon , Ryong Lee Byung , Jin Li Hua , Park Jinseu , Choi Soo Young
Published: July 2002

A human Cu,Zn‐superoxide dismutase (Cu,Zn‐SOD) gene was fused with a gene fragment encoding the nine amino acid transactivator of transcription (Tat) protein transduction domain (RKKRRQRRR) of HIV‐1 in a bacterial expression vector to produce a genetic in‐frame Tat–SOD fusion protein. The expressed and purified Tat–SOD fusion protein in Escherichia coli can enter HeLa cells in a time‐ and dose‐dependent manner when added exogenously in a culture media. Denatured Tat–SOD protein was transduced much more efficiently into cells than were native proteins. Once inside the cells, transduced Tat–SOD protein was enzymatically active and stable for 24 h. The cell viability of HeLa cells treated with paraquat, an intracellular superoxide anion generator, was increased by transduced Tat–SOD. These lines of results suggest that the transduction of Tat–SOD fusion protein may be one of the ways to replenish the Cu,Zn‐SOD in the various disorders related to this antioxidant enzyme.
A docking model of key components of the DISC complex: death domain superfamily interactions redefined
Authors: Weber Christian H , Vincenz Claudius
Published: July 2002

Apoptosis is mediated by a highly regulated signal transduction cascade that eventually leads to precisely directed cell death. The death‐inducing signaling complex (DISC), composed of Fas, FADD, and caspase‐8, is an apical signaling complex that mediates receptor‐induced apoptosis. We have docked the experimentally determined structures of the Fas and FADD death domains into a model of a partial DISC signaling complex. The arrangement of Fas and FADD was determined using the interaction modes of the two heterodimer crystal structures determined to date, Pelle/Tube and Apaf‐1/procaspase‐9. The proposed model reveals that both interactions can be accommodated in a single multimeric complex. Importantly, the model is consistent with reported site‐directed mutagenesis data indicating residues throughout the domain are critical for function. These results imply that members of the death domain superfamily have the potential for multivalent interactions, offering novel possibilities for regulation of apoptotic signaling.
The site of production of superoxide radical in mitochondrial Complex I is not a bound ubisemiquinone but presumably iron–sulfur cluster N2
Authors: Genova Maria Luisa , Ventura Barbara , Giuliano Giovanni , Bovina Carla , Formiggini Gabriella , Parenti Castelli Giovanna , Lenaz Giorgio
Published: July 2002

The mitochondrial respiratory chain is a powerful source of reactive oxygen species, considered as the pathogenic agent of many diseases and of aging. We have investigated the role of Complex I in superoxide radical production in bovine heart submitochondrial particles and found, by combined use of specific inhibitors of Complex I and by Coenzyme Q (CoQ) extraction from the particles, that the one‐electron donor in the Complex to oxygen is a redox center located prior to the binding sites of three different types of CoQ antagonists, to be identified with a Fe–S cluster, most probably N2 on the basis of several known properties of this cluster. Short chain CoQ analogs enhance superoxide formation, presumably by mediating electron transfer from N2 to oxygen. The clinically used CoQ analog, idebenone, is particularly effective in promoting superoxide formation.
Degradation of tyrosine aminotransferase (TAT) via the ubiquitin–proteasome pathway
Authors: Gross-Mesilaty Shlomit , Hargrove James L , Ciechanover Aaron
Published: July 2002

Most of the known cellular substrates of the ubiquitin system are short‐lived growth regulators and transcriptional activators. Very few enzymes involved in intermediary metabolism have been shown to be targeted by the system. In a reconstituted cell‐free system, we show that tyrosine aminotransferase (TAT), a key enzyme involved in amino acid metabolism, is conjugated and degraded in an ATP‐ and ubiquitin‐dependent manner. Degradation of ubiquitin–TAT adducts requires, in addition to the 26S proteasome, a novel, yet unidentified, factor. TAT can be protected from degradation by association with its coenzyme pyridoxal phosphate. To examine the potential role of the ubiquitin system in regulating the stability of the enzyme in vivo, we show that cell extracts derived from livers of animals in which TAT was induced, display a corollary increase in the formation of specific TAT–ubiquitin adducts. © 1997 Federation of European Biochemical Societies
Mouse muscle regeneration: an in vivo 2D 1H magnetic resonance spectroscopy (MRS) study
Authors: Sébrié C , Gillet B , Lefaucheur J.P , Sébille A , Beloeil J.C
Published: July 2002

Muscle degeneration and regeneration were studied by 2D 1H magnetic resonance spectroscopy (MRS) and histological examination, in an experimental model of muscle injury using a myotoxic snake venom, notexin. The injured muscles produced a very specific MRS signal, corresponding to a tri‐unsaturated fatty acid (linolenic acid‐like) signal, from day 2 to day 9 after injury. The combination of MRS with histology showed that this signal was associated with a mechanism occurring during myoblast fusion to form myotubes. 2D 1H MRS is thus a useful non‐invasive tool for detecting muscle regeneration in vivo.
SKAP‐HOM, a novel adaptor protein homologous to the FYN‐associated protein SKAP551
Authors: Marie-Cardine Anne , Verhagen Anne M. , Eckerskorn Christoph , Schraven Burkhart
Published: July 2002

A recombinant GST‐Fyn‐SH2 domain was used to purify proteins from lysates of pervanadate treated EL4 cells. N‐terminal sequencing and molecular cloning of one of the purified polypeptides resulted in the identification of a novel adaptor protein that shares strong structural homology to the recently cloned Fyn‐associated adaptor protein SKAP55. This protein was termed SKAP‐HOM (55 ologue). Despite their striking homology, SKAP55 and SKAP‐HOM have distinct characteristics. Thus, unlike SKAP55, which is exclusively expressed in T lymphocytes, SKAP‐HOM expression is ubiquitous. Furthermore, while SKAP55 is constitutively tyrosine phosphorylated in resting human T cells, SKAP‐HOM is expressed as a non‐phosphorylated protein in the absence of external stimulus but becomes phosphorylated following T cell activation. In addition, SKAP‐HOM does not associate with p59fyn in T cells although it represents a specific substrate for the kinase in COS cells. Finally, we demonstrate that, as previously shown for SKAP55, SKAP‐HOM interacts with the recently identified polypeptide SLAP‐130.
Differential effects of alkaloids on sodium currents of isolated single skeletal muscle fibers
Authors: Körper Sixten , Wink Michael , Fink Rainer H.A
Published: July 2002

The effects of the alkaloids ajmaline, lupanine, sparteine, serpentine, strychnine, and yohimbine were studied with the loose patch clamp technique on sodium currents of isolated single skeletal muscle fibers. The IC50 values for half‐maximal blocking of the sodium currents were 6.6 μM for ajmaline, 55.7 μM for quinidine, 168.8 μM for sparteine, and 1.2 mM for lupanine. The observed Na+ channel inhibition is in accordance with the use of ajmaline, quinidine and sparteine as antiarrhythmic drugs. The interference of alkaloids with Na+ channels can also be interpreted as a means to strongly interfere with neuronal transmission in herbivores. Alkaloids thus serve as chemical defense compounds for the plants producing them.
Characterization of a cDNA encoding a precursor of Carassius RFamide, structurally related to a mammalian prolactin‐releasing peptide
Authors: Satake Honoo , Minakata Hiroyuki , Wang Xiaoyan , Fujimoto Masaaki
Published: July 2002

We have characterized the cDNA encoding Carassius RFamide (C‐RFa), which is structurally related to mammalian prolactin‐releasing peptides (PrRPs), from the brain of the crucian carp. The deduced C‐RFa precursor has been shown to comprise 117 amino acids, encoding a single C‐RFa sequence. A comparative study of amino acid sequences has revealed that several sequences conserved in preproPrRPs are also found in the C‐RFa precursor. Furthermore, the abundant presence of the C‐RFa mRNA in the telencephalon, optic tectum, medulla oblongata, and proximal half eye ball was demonstrated by Southern blot analysis of RT‐PCR products.
Distinct roles of profilin in cell morphological changes: microspikes, membrane ruffles, stress fibers, and cytokinesis
Authors: Suetsugu Shiro , Miki Hiroaki , Takenawa Tadaomi
Published: July 2002

Here we report the functional importance of profilin in various actin‐mediated morphological changes using H119E mutant profilin I, which is deficient only in actin binding. In the case of actin‐protrusive structures from the plasma membrane, H119E‐profilin was shown to suppress the formation of Cdc42‐induced actin microspikes and Rac‐induced membrane ruffles. Conversely, Rho‐induced stress fiber formation seemed to occur independently of H119E‐profilin introduction. Furthermore, H119E‐profilin blocked cleavage furrow ingression and subsequent adhesion to the substratum during cell division, a process in which actin plays indispensable roles.
Increase of urinary extracellular‐superoxide dismutase level correlated with cyclic adenosine monophosphate
Authors: Adachi Tetsuo , Yamada Harutaka , Hara Hirokazu , Futenma Arao , Kakumu Shinichi
Published: July 2002

Extracellular superoxide dismutase (EC‐SOD) is a secretory protein that is the major SOD isozyme in extracellular fluids. Plasma EC‐SOD levels are distributed in two discrete groups with the rare group having an enzyme with glycine instead of arginine‐213, which causes a 10‐fold higher serum level. Within the common phenotype group, the urinary EC‐SOD level was significantly correlated with the urinary excretion of N‐acetyl‐β‐D‐glucosaminidase (NAG), but not with serum EC‐SOD. EC‐SOD appears not to be leaked from the plasma by glomerular filtration, but rather to be secreted from the renal tubule or its surrounding tissues. The urinary EC‐SOD level was also significantly correlated with the urinary cyclic adenosine monophosphate (cAMP) level. cAMP analogues and adenylate cyclase modulators significantly stimulated the expression of EC‐SOD but not other SOD isozymes in cultured fibroblast cell lines. Moreover, injection of parathyroid hormone, in Ellsworth‐Howard tests, increased urinary EC‐SOD accompanied with the elevations of urinary cAMP and NAG. Together these observations suggest that factor(s) that stimulate the adenylate cyclase‐cAMP system regulate the urinary EC‐SOD level.
Malaria prophylaxis and the reduction of anemia at childbirth
Authors: Geelhoed D.W , Visser L.E , Addae V , Asare K , Schagen van Leeuwen J.H , van Roosmalen J
Published: July 2002

Objectives: To assess the effect of introducing chloroquine prophylaxis during pregnancy on prevalence of anemia (<10.9 g/dl) at childbirth and perinatal outcome. Methods: Observational study in a rural district hospital in Ghana, which compared 2803 women who received chloroquine prophylaxis during pregnancy with 3084 historical controls, who had not received prophylaxis during pregnancy. Main outcome measures were hemoglobin level at childbirth, perinatal mortality and birth weight. Results: Mean hemoglobin level before childbirth increased from 10.7 g/dl (S.D.=1.0 g/dl) to 11.0 g/dl (S.D.=0.9 g/dl). Prevalence of anemia decreased from 29.4 to 13.3% (OR=0.4 and 95% CI=0.3–0.4). Prevalence of moderately severe anemia (<9.0 g/dl) decreased from 4.4 to 3.3% (OR=0.7, 95% CI=0.6–0.97). Perinatal mortality and low birth weight (<2500 g) remained unchanged. Conclusions: Routine chloroquine prophylaxis in pregnancy is useful in reducing anemia at childbirth in malaria‐endemic regions. Fetal outcome did not improve with chloroquine prophylaxis in this study.
Caspase‐3 protease activation during the process of genistein‐induced apoptosis in TM4 testicular cells
Authors: Kumi‐Diaka James , Butler Andre
Published: July 2002

Summary— The role of caspase‐3 (CPP32) protease in the molecular pathways of genistein‐induced cell death in TM4 cells was investigated. Fluorescence microscopy with Hoechst‐33258‐PI nuclear stain was used to distinguish between apoptosis and necrosis pathways of cell death. The viability of the test cells was assessed with both the trypan blue exclusion and MTT tetrazolium (3‐[4,5‐dimethyl‐thiazol‐2‐yl]‐2,5‐diphenyltetralzolium bromide, 2.5 mg/mL) assays. Caspase‐3 enzymatic activity was determined using CasPASE Apoptosis Assay Kit. The overall results from all the data demonstrated that: i) genistein exerts dose‐ and time‐dependent effects on TM4 testis cells; ii) apoptosis is induced by lower concentrations of genistein and necrosis induced by higher concentrations of genistein; iii) genistein induced activation caspase‐3 enzymatic activity; iv) genistein‐induction of apoptosis and necrosis was significantly inhibited by the caspase‐3 inhibitor, z‐DEV‐FMK; v) sodium azide induced necrosis without activation of CPP32 enzymatic activity, and induction of apoptosis; and vi) genistein‐induced apoptosis was associated with activation of CPP32 enzymatic activity in the cells. The overall results indicate a strong evidence of caspase‐3 (CPP332) mediation in the molecular pathways of genistein‐induced apoptosis in testicular cells. Apoptosis is the physiologically programmed cell death in which intrinsic mechanisms participate in the death of the cell, in contrast to necrosis, which induces inflammatory response in the affected cell. The fact that the chemopreventive role of several cancer drugs is due to induction of apoptosis augments the biotherapeutic potential of genistein for the treatment of malignant diseases including prostate and testicular cancers. It is therefore inevitable that identification of the apoptotic pathways and the points at which regulation occurs could be instrumental in the design of genistein biotherapy for such diseases.
Novel genes expressed in the chick otocyst during development: Identification using differential display of RNA
Authors: Gong Tzy‐Wen L. , Hegeman Adrian D. , Shin Jooyoung J. , Lindberg Kendra H. , Barald Kate F. , Lomax Margaret I.
Published: July 2002

Differential display of mRNA is a technique that enables the researcher to compare genes expressed in two or more different tissues or in the same tissue or cell under different conditions. The method is based on polymerase chain reaction amplification and comparison of specific subsets of mRNA. We have used this method to clone partial complementary DNAs (cDNAs; amplicons) for genes expressed in the otocyst in order to identify genes that may be involved in development of the inner ear. A full length cDNA was isolated from an embryonic quail head library with an amplicon (KH121) obtained from the otocyst. This avian cDNA encoded a novel, 172‐amino acid acidic protein and detected a major transcript of ca 0.8 kb in RNA from chick embryos and several neonatal chick tissues. The full length avian cDNA had high sequence identity to several human cDNAs (expressed sequence tags) from human fetal tissues, including cochlea, brain, liver/spleen and lung, and from placenta. The human homologue of the avian gene encoded a protein that was 183 amino acids long and had 75.6% amino acid sequence identity to the avian protein. These results identified both the avian and human homologues of an evolutionarily conserved gene encoding a small acidic protein of unknown function; however, expression of this gene was not restricted to otocysts.
Age‐dependent changes of presynaptic neuromodulation via A1‐adenosine receptors in rat hippocampal slices
Authors: Sperlágh B. , Zsilla G. , Baranyi M. , Kékes‐Szabó A. , Vizi E.Sylvester
Published: July 2002

Thepresynaptic neuromodulation of stimulation‐evoked release of [3H]‐acetylcholine by endogenous adenosine, via A1‐adenosine receptors, was studied in superfused hippocampal slices taken from 4‐, 12‐ and 24‐month‐old rats. 8‐Cyclopentyl‐1,3‐dimethylxanthine (0.25 μM), a selective A1‐receptor antagonist, increased significantly the electrical field stimulation‐induced release of [3H]‐acetylcholine in slices prepared from 4‐ and 12‐month‐old rats, showing a tonic inhibitory action of endogenous adenosine via stimulation of presynaptic A1‐adenosine receptors. In contrast, 8‐cyclopentyl‐1,3‐dimethylxanthine had no effect in 24‐month‐old rats. 2‐Chloroadenosine (10 μM), an adenosine receptor agonist decreased the release of [3H]‐acetylcholine in slices taken from 4‐ and 12‐month‐old rats, and no significant change was observed in slices taken from 24‐month‐old rats. In order to show whether the number/or affinity of the A1‐receptors was affected in aged rats, [3H]‐8‐cyclopentyl‐1,3‐dimethylxanthine binding was studied in hippocampal membranes prepared from rats of different ages. Whereas the Bmax value was significantly lower in 2‐year‐old rats than in younger counterparts, the dissociation constant (Kd) was not affected by aging, indicating that the density rather than the affinity of adenosine receptors was altered. Endogenous adenosine levels present in the extracellular space were also measured in the superfusate by high performance liquid chromatography (HPLC) coupled with ultraviolet detection, and an age‐related increase in the adenosine level was found.

Insummary, our results indicate that during aging the level of adenosine in the extracellular fluid is increased in the hippocampus. There is a downregulation and reduced responsiveness of presynaptic adenosine A1‐receptors, and it seems likely that these changes are due to the enhanced adenosine level in the extracellular space.
Inter-rater reliability of ICD-10 in Chinese patients
Authors: Pang AHT , Ungvari GS , Wong CK , Leung T
Published: July 2002

In an attempt to assess the universal applicability of the International Classification of Disease (ICD-10), two psychiatrists from different socio-cultural backgrounds and training independently performed a chart review of 238 Chinese patients. Inter-rater reliability figures were comparable to those found in the WHO-coordinated ICD-10 field trials. The results suggest that ICD-10 has good ‘universality’ in routine clinical practice.
Online Women's Networks
Authors: Kannengießer Sigrid
Published: July 2020

The entry describes the phenomenon of online women's networks and sketches out key findings of the research field in gender media studies which analyzes how women and women's organizations use online media for networking, mobilization, and articulation. Different forms of online women's networks are taken into account: online women's networks which are established spontaneously through online networking sites such as Twitter and Facebook because of a special event or incident as well as women's organizations which organize and communicate via online media are considered. With the internet, women and women's organizations acquired new ways of connecting, mobilizing, and articulating their interests and demands. But at the same time mediated communication enabling the networking among women and women's organizations produces unequal power relations within the networks and between the networks and their environments.
Pro‐Ana and Thinspiration
Authors: Ging Debbie
Published: July 2020

Pro‐anorexia is a collective response to the eating disorder anorexia, which emerged on the internet in the early 1990s. Thinspiration refers to a similar—though more mainstream—phenomenon, whereby social media network users share dieting and weight‐loss tips and images. People with bulimia and EDNOS (eating disorders not otherwise specified) are also included in this community, with the former frequently referred to as “Mia.” It is estimated that 90–95% of the sufferers of anorexia and bulimia are women. The pro‐ana community typically rejects the medical diagnosis of eating disorders as an illness, arguing that food restriction and weight loss are a lifestyle choice and part of their identity.
Slash
Authors: Busse Kristina
Published: July 2020

Slash describes creative fan works featuring same‐sex romantic and sexual relationships, often expanding on homosocial bonds in the source texts. Broadly, it can be used as a general descriptive genre of homoerotic transformative art, but more commonly it means a specific historical, social, and economic category connected to media fandom, the creative primarily female fan communities of live‐action Western TV shows from the 1960s onward. Slash fan fiction has been a central focus of fan and audience studies: its primarily female readers and writers as well as its explicit homoerotic prose defies easy explanations of women's genres and media receptions. Moreover, its often‐aggressive alternate readings of popular media texts exemplify reception models of resistant readings. Following greater visibility of queer characters and relationships on screen, slash continues to be popular especially in its exploration of non‐normative sexualities and sexual identities.