bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Cooking Methods on the Nutritive Value and Lipid Oxidation of Two Cricket Species Consumed In Cameroon
Authors: Tenyang Noel , Tiencheu Bernard , Mamat Abazidi , Mawamba Ludovine Ateufack , Ponka Roger
Published: March 2021

Aims: The present study was performed in order to evaluate the effect cooking on the proximate composition and lipid oxidation of two cricket species (Hieroglyphus daganensis and Paracinema tricolor) commonly consumed in Cameroon.
Study Design: Paracinema tricolor and Hieroglyphus daganensis harvesting, cleaning, killing, cooking and drying, evaluation the effect cooking treatment on the nutritive value and lipid oxidation of these crickets species.
Place and Duration Study: University of Maroua, Cameroon and University of Yaounde I in Cameroon from November 2018 to July 2019.
Methodology: The two types of crickets after harvesting and transporting to the laboratory, were killed by freezing at -20°C for 24 hours. Common cooking methods (pan roasting and pan frying) were used to cook it. The raw, roasted and fried crickets were left at ambient temperature and then one part was transformed into powder using an electric grinder. Before analysis, the powders and the whole samples were stored at 4°C. The ash, lipid, protein, carbohydrates and crude fibber content were determined in powder samples. The oil was extracted from whole samples of raw, roasted and fried insects. Various chemical parameter (acid value, peroxide value, iodine value and thiobarbituric acid) were used to assess crickets’ lipid qualities during the cooking methods.
Results: The proximate analysis of raw samples revealed that the ash content of Paracinema tricolor and Hieroglyphus daganensis was 3.17% and 3.49% (% of dry weight) respectively. The lipid content was generally high ranging from 15.80% to 18.10% and the protein content ranged from 41.69% to 48.59%. The raw Paracinema tricolor compared to Hieroglyphus daganensis was found to have the higher contents of lipid, protein and energy.  All the cooking methods increased the lipid, protein and ash contents, as well as the calorie values of the two cricket’s species, while the moisture content was reduced. The free fatty acids and thiobarbituric acid reactive substances (TBARS) values of the two crickets’ oil were significantly increased after roasting and frying processing. Iodine value decreased.
Conclusion: From these results, it can be concluded that soaking combined with frying method appeared to be the best cooking method in the two species of crickets in terms of caloric content and lipid oxidative stability.
Effects of a Fraction of the Methanol Extract of the Seeds of Abrus Precatorius on Malondialdehyde and Antioxidant Levels of Alloxan-Induced Diabetic Rats
Authors: Iloanya Ebele Lauretta , Nwodo Okwesili Fred Chiletugo , Ezeigwe Obiajulu Christian , Alaebo Ogochukwu Prince , Enemchukwu Nnanyelugo Benneth , Aziagba Okwuchukwu Bibian , Okpani Ifeanyi Sunday
Published: March 2021

Introduction: The aim of this study is to evaluate the effect of fraction 2 (F2) of methanol extract of the seeds of Abrus precatorius on malondialdehyde and antioxidant level of alloxan-induced diabetic Wistar albino rats.

Materials and Methods: The methanol extract of the seeds of A. precatorius Linn Fabaceae was fractionated by Sephadex G15. Diabetes was induced by a single intraperitoneal administration of alloxan at a dose of 150 mg/kg bodyweight. The phytochemical analysis and the biochemical parameters were investigated using standard diagnostic methods.

Results: Both qualitative and quantitative analysis of F2 revealed the presence of high amounts of alkaloids (2000 ± 80 mg/100 g), flavonoids (158 ± 17.6 mg/100 g) and tannins (258 ± 45 mg/100 g) but low concentration of saponins (18.3 ± 2.43 mg/100 g). The malondialdehyde (MDA) level of all the test groups and the group treated with the standard drug decreased significantly (p<0.05) compared with that of untreated diabetic group. However, the catalase activity in all the test groups significantly increased (p<0.05) while the activity of superoxide dismutase increased significantly (p<0.05) in groups administered 20mg/kg of the fraction and the group pre-treated with 10mg/kg of the fraction compared with the diabetic untreated group. Similarly, the result revealed a significant increase (p<0.05) in non-enzymatic antioxidants such as reduced glutathione and vitamin E of the rats treated with graded doses of F2 while vitamin C showed significant (p<0.05) increase in normal control groups and pre-treated groups compared with the diabetic untreated. Studies on membrane stabilization using hypotonicity-induced red blood cell haemolysis revealed that the F2 of the methanol extract of the seeds of A. precatorius Linn inhibited haemolysis in a dose-dependent manner.

Conclusion: The results obtained from this study revealed that the F2 fraction of the methanol extract of A. precatorius contain important phytochemicals found in the Fraction 2 (F2) however showed improvement in antioxidant defence as well as has a stabilizing effect on the membrane.
Probiotic Potential and Genomic Evaluation of Lactic Acid Bacteria Isolated from Fermented Sorghum-Based Gruel
Authors: Popoola Oluwabukola Atinuke , Onilude Abiodun A. , Rasheed-Jada Hadiza , Nashiru Oyekanmi
Published: March 2021

Forty eight isolates were randomly isolated from fermented red sorghum (Sorghum bicolor) and white sorghum-based (Sorghum vulgare) gruel and evaluated for their probiotic abilities. Forty five out of the forty eight isolates were Gram-positive, catalase negative and non-sporulating, which were further evaluated.  Twenty out of the forty five isolates showed tolerance to acid with their ability to survive extreme pH 2.0, 3.0 and 4.0 respectively at different levels. These were further screened for their in vitro inhibition ability against selected test pathogens (Escherichia coli, Salmonella Typhimurium, Bacillus sp., Pseudomonas aeruginosa, and Staphylococcus aureus). Fourteen out of the twenty isolates had antimicrobial activity against the test pathogens at different levels. They were further screened for their antibiotic susceptibility pattern, all the fourteen isolates showed susceptibility to different antibiotics at different levels. These isolates were screened for their bile tolerance ability, all the fourteen isolates showed tolerance to the bile salt concentration (0.3%, 0.5% and 1%) at different levels respectively. The fourteen isolates were also screened for their gastric transit tolerance ability; all the fourteen isolates were able to tolerate gastric transit at 0 minute, 90 minutes and 180 minutes respectively. They were further screened for their in vitro hydrophobicity ability, only six out of the fourteen isolates showed percentage hydrophobicity of 40% and above. These six isolates were identified at the sub-species level using 16S RRNA Sequencing for isolates identification and construction of phylogeny. They were identified as Lactobacillus plantarum strain ST1, Lactobacillus brevis strain SC4, Lactobacillus casei strain 0108, Lactobacillus plantarum strain HASOB9a, Pediococcus pentosaceus strain SH 740 and Lactobacillus plantarum ZJ316 respectively. Out of the six identified isolates, the complete genome of Lactobacillus plantarum ZJ316 had been sequenced and was retrieved alongside with the complete genome data of four other species of Lactobacillus plantarum of industrial importance available in public domain, which was used for the mini comparative genome analysis in this research, using bioinformatics and visual analytics tools (Tableau Desktop). It was revealed that the genome size of Lactobacillus plantarum ZJ316 is second to that of Lactobacillus plantarum 16 and higher than that of the others whose genome had been completely sequenced.
Evalution of Human Amniotic Membrane in Extraction Socket for Faster Wound Healing
Authors: Kumar S. P. Indra , Fathima Afrose , Deepika S. , Aravind R. J. , Kavin T. , Priyadharshini Gayathri , Narendar R.
Published: March 2021

Background: Preservation of extraction site with grafting shall always produce better healing results. Various materials have been used for this process and each material has its unique benefits and properties. Few such materials are Simvastatin, Bone ceramic, oxidized cellulose, bone allograft, etc. In this given study, 10 patients were placed Human amniotic membrane in their extracted socket and the results were reviewed after 7days, 14 days, 21 days, 1 month, 3 month and 6 month.

Materials and Methods: The samples are voluntary and those patients with no clinical abnormalities were included. Human amniotic membrane is received from Tata Memorial Hospital, Mumbai.

Ten patients who reported to Department of OMFS were selected for the augmentation procedure. After extraction, the Human amniotic membrane was placed in the extracted socket and secured with sutures. Post-operative visits scheduled after 7days, 14 days, 21 days, 1 month, 3 month and 6 month intervals for analyzing the he.0aling property of amnion in the augmented extraction sockets.

Results: The results of this study have shown that Amniotic membrane has proven its effect, as it initiates faster wound healing of the extraction socket.

Conclusion: This article shall reveal the effectiveness of the Human amniotic membrane in rapid wound healing of the augmented extraction socket.
Personal Attribution of Millennial Generation: Contribution of Communication Style and Personality to Friendship Quality at BPK PENABUR Bandung
Authors: Rewindinar . , Marta Rustono Farady , Tjajadi Octavia Putri , Latukolan Jovid Jonathan
Published: March 2021

Technology plays a major role in the activities of the millennial generation and is a characteristic of this productive age generation. This study stems from the assumptions of the millennial generation who were present at a time of widespread technological development. The millennial generation has communication problems that are inherent in personality. This study questioned how far the contribution of communication style and personality to friendship quality, in this case, the factors that influence individual behavior. This study aimed to see the contribution between communication style and personality with friendship quality. This study used Attribution Theory which looks at the factors affecting the behavior of others, which are situational and personality factors. The study was conducted using a quantitative methodology with a survey method. The online survey was conducted on 114 participants consisting of teachers and employees at BPK PENABUR Bandung. The participants are the millennial generation who were born between the years of 1981-1995. Data analysis used multiple regression with P <.01. The results show a moderate correlation (r = .463) between communication styles and friendship quality. Also, it shows a moderate correlation (r = .514) between the personality types and friendship quality. The contribution of the communication style and personality variables to the friendship quality is 28.5%. This study describes communication styles and personality as fading personal factors in affecting the level of depth of relationships among the millennial generation.
Distributed Leadership in Greek Public Schools: Limits and Conditions
Authors: Gkoros Dimitrios
Published: March 2021

Aim: To know whether the legal and institutional framework of the educational structure and operation, therefore of the school leadership as follows, is properly formed in Greek educational system and what the model of distributed leadership plays in the Greek educational context.

Results: The organization of Greek formal education is based on the bureaucratic system. This centralized system of school organization leaves little room for action and initiative for principals and, consequently, for teachers. It is important that the personal vision of the principals is transmitted to the other members of the school, in possible sources of leadership, as a factor of change and inspiration for all, overcoming the abovementioned bureaucratic system but also for the system itself to change and become more diverse and flexible.

Conclusion: In our opinion the detachment from the traditional, hierarchical forms of leadership and the transition to the division of responsibilities and the decentralization of power is imperative. Despite the typical structure of any education system, we believe that schools maintain some degree of autonomy that they can use to create a power distribution network, even informally.
Evaluation of Vitamin Contents, Antioxidant and Antimicrobial Activities of Different Leaf Extracts of Taraxacum officinale (Dandelion)
Authors: Daniel I. E. , Mathew K. N. , John P. L.
Published: March 2021

Methanol and ethyl acetate leaf extracts of Taraxacum officinale (dandelion) were evaluated for phytochemical compounds, vitamins, antioxidant and antimicrobial activities. Phytochemical compounds and vitamins were determined using standard procedures while antioxidant activity was determined using 2, 2’-diphenyl-1-picrylhydrazyl (DPPH) and Ferric reducing/antioxidant power (FRAP) Assay.  Antimicrobial activity against various pathogenic bacteria and fungi were screened using disc diffusion method. The results indicated that the bioactive compounds (total phenol, flavonoids, saponins, tannins and alkaloids) determined quantitatively were present in appreciable concentration in both extracts. The result also showed that both extracts contain a variety of vitamins (A, B complex, C and E), with vitamins C and A having the highest concentration while the B-vitamins (B1, B2 and B3) and vitamin E were present in moderate concentrations. Both extracts showed significant scavenging and reducing ability comparable to the reference antioxidant, ascorbic acid in a dose dependent manner, with methanol exhibiting the highest scavenging and reducing capacity. The antimicrobial activity of both extracts showed appreciable broad spectrum activity against the pathogenic bacteria and fungi strains tested at various concentrations. Methanol extract was found to be most effective compared to ethyl acetate extract. These results indicated that the leaf extracts of dandelion possess antioxidant and antibacterial activity against the tested bacteria possibly due to the presence of bioactive compounds and other nutrients.
Detection of INVA Gene and Cytotoxin of Salmonella entertidis in Food Samples Using Molecular Methods
Authors: Almas Maha , Tahir Usaal , Zameer Mariam , Shafiq Muhammad Imtiaz , Farrukh Syeda Yumna , Zahra Nureen , Mazhar Maham , Gill Sajjad Ali , Hadi Faheem , Muhammad Tahir , Ali Qurban , Malik Arif
Published: March 2021

Salmonella entertidis is a foodborne pathogen that causes various diseases in human beings worldwide. The toxin of Salmonella can cause infectious diseases. In this research project, Salmonella was detected through various microbial, biochemical and molecular tests in diverse food samples collected from highly populated, moderately populated and less populated areas of Lahore, Pakistan. Enriched cultures of all food samples such as apples, tomatoes, yogurt and mayonnaise was streaked on violet-red bile glucose agar, Simmon’s citrate agar and eosin-methylene blue agar (EMB).  Salmonella isolates were screened for the presence of toxin encoding gene through PCR. 27% apples, 19% tomatoes, 5% mayonnaise and 7% yogurt were found to be positive for INVA genes (invasion protein genes). In medical and pharmaceutical point of views the INVA gene can also help to develop specific medicines against salmonella. The cytotoxin that is protein in nature was confirmed by SDS PAGE in mayonnaise samples. This study illustrates that foods of highly populated areas are reservoir for Salmonella entertidis in Pakistan. There is need to develop specific drugs, precautionary measures to control salmonella and its disease.
Analysis of the Range of Medicines used in the Treatment of Skin Diseases
Authors: Akhtaev Ibragim Khalidovich , Romantseva Elizaveta Vitalievna , Vasiliev Danila , Kulish Anastasia , Altemirova Mata Musaevna , Goncharov Vitaly V.
Published: March 2021

The purpose of this article is to analyze the range of medicines that are used in the treatment of skin diseases. This topic is currently relevant, since the analysis of retail sales allows you to assess the financial condition of a pharmacy organization, identify unclaimed drugs, develop measures to stimulate sales, as well as study customer demand and develop a strategy for product movement. Accordingly, conducting a study of the main indicators of the assortment of this group of drugs will make it possible to expand their representation on the shelves of pharmacies, as well as to study the demand for them from potential consumers.

During the analysis, the following data were obtained. The range under study was represented in the pharmacy by 22 items. The analysis of dosage forms of medicines intended for the treatment of skin diseases showed that liquid dosage forms accounted for 22.73%; solid dosage forms - 4.55%; soft dosage forms – 59.08%; gaseous dosage forms -13.64%. When analyzing the drugs, it was revealed that the largest number in the assortment is synthetic drugs - 68.18%; the share of phytopreparations was 31.82%. Analysis of the producing countries showed that most of the drugs presented to the pharmacy by domestic producers (54,53%), in the second place drugs manufactured in India -18,17%; third place was shared by countries: Poland, Croatia, Belgium, Germany, Czech Republic, USA -4,55%. The analysis of price categories of pharmaceuticals for the treatment of skin diseases revealed that in the price category from 100 to 200 RUB presents 22,73% of drugs; in the category from 200 to 300 rubles-49,89%; from 300 to 400 rubles from 33.73%; from 400 to 700 rubles - on 4,55%.
Medication Errors and Pharmacist Intervention at Government Hospital of Hyderabad, Pakistan
Authors: Saleem Raheela , Dayo Abdullah , Ghoto Muhammad Ali
Published: March 2021

Background: Medication errors are of primary concern in hospitals of Pakistan but only few are reported. Prevalence and type of medication errors are essential for estimating the appropriateness of therapy. Pharmacist is key in improving therapy effectiveness and decreasing medication errors.

Objective: To assess the prevalence, type of medication errors and pharmacist interventions.

Methodology: An interventional study on prescriptions of inpatients was conducted at Liaquat University of Medical and Health Sciences (LUMHS) Hyderabad. Five hundred and fifteen prescriptions were collected randomly from medicine, gynecology and obstetrics and urology department during one year from 01-09-2017 to 31-08-2018. Medications errors were identified by Pharmacist by comparing the prescriptions with standard of British National Formulary (BNF) (edition 73) and informing physician for resolving medication errors. Data was analyzed with statistical package for social sciences version 25.0.

Results: Out of 515 inpatients prescriptions, male patients were 248 (48.2%) and female were 267 (51.8%) with male to female ratio of 1:1.08 and mean age of 37.7 ± 11.6 (18-60) years. 427 medication errors were reported in 303 (59.4%) prescriptions including; omission error 64 (15.0%), unauthorized drug error 41 (9.6%), wrong dose error 67 (15.7%), wrong time error 74 (17.3%), wrong dosage form error 27 (6.3%), wrong route error 23 (5.4%), wrong drug preparation 44 (10.3%), wrong rate error 56 (13.1%) and incorrect administration technique 31 (7.3%). 172 (40.3%) interventions of pharmacists were accepted by physicians.

Conclusion: Rate of medication errors in inpatients prescriptions are high with possible errors of dose, time, omission, rate, drug preparation and others. Pharmacist effectively intervening the prescriptions and playing active role in decreasing medication errors.
Chemical shifts of aromatic protons in protein NMR spectra
Authors: Redfield Christina , Hoch Jeffrey C. , Dobson Christopher M.
Published: July 2002

Experimental secondary shifts have been compared with calculated ring current shifts for the aromatic protons of lysozyme. The root mean square differences were calculated for all possible permutations of the assignment scheme. The experimentally determined assignment scheme was found to give the best agreement with the ring current shift calculations. Similar results were obtained in the case of bovine pancreatic trypsin inhibitor and cytochrome c. The use of the degree of correlation between experimental and predicted NMR parameters as a basis for making assignments is examined.
Amino acid sequence characterization of mammalian vimentin, the mesenchymal intermediate filament protein
Authors: Geisler Norbert , Plessmann Uwe , Weber Klaus
Published: July 2002

The amino‐terminal 98 residues of porcine vimentin have been determined by amino acid sequence studies. Extensive overlap is seen with the corresponding region of the carboxyterminal 448 residues of hamster vimentin predicted from DNA sequence studies, which left the very amino‐terminal region unknown. The combined data show that contrary to gel electrophoretic results, mammalian vimentin contains only about 467 residues, and that species‐specific drift occurs mainly in the amino‐terminal non‐α‐helical array. The results are discussed parallel to emerging concepts on intermediate filament protein diversity.
A new method of the measurement of the electrically neutral fluxes of cations through lipid bilayer membranes induced by Men+/nH+‐exchangers
Authors: Antonenko Yury N. , Yaguzhinsky Lev S.
Published: July 2002

Electrically neutral ionophores (nigericin, monencin) incorporated into a planar bilayer lipid membrane (BLM) bring about hydrogen ion gradient formation in the unstirred layers of BLM if a metal ion gradient on the membrane is prepared. Under these conditions a diffusion potential of a hydrogen ion is generated after addition of a protonophore. Cation selectivity of nigericin, monencin and A23187 has been studied by means of electrical potential measurements in the presence of a protonophore and Me
n+/nH+‐exchangers mentioned above. The data on cation selectivity are in a good agreement with the well known results of the direct measurements of metal ion fluxes. This shows that the effect of generation of the potential on BLM in the presence of a protonophore and a Me
n+/nH+‐exchanger can be used for the estimation of electrically neutral ion fluxes through BLM.
Complete amino acid sequence of human seminal plasma β‐inhibin
Authors: Seidah N.G. , Arbatti N.J. , Rochemont J. , Sheth A.R. , Chrétien M.
Published: July 2002

The complete sequence of a 94 amino acid human seminal plasma polypeptide exhibiting inhibin‐like activity is presented. This molecule, called β‐inhibin, selectively and specifically suppresses the release of pituitary FSH in vivo as well as in vitro. It does not affect the secretion of LH. Such a novel acidic protein contains a very basic C‐terminal segment which is easily cleaved by mild tryptic digestion. It is predicted that the FSH inhibiting activity may reside within this region of the molecule. This would imply a post Gin‐Arg cleavage to release the basic C‐terminal active moiety.
Mitomycin C‐induced deoxyribose degradation inhibited by superoxide dismutase
Authors: Gutteridge John M.C. , Quinlan Gregory J. , Wilkins Stephanie
Published: July 2002

Mitomycin C stimulates deoxyribose degradation with the release of thiobarbituric acid‐reactive material under conditions of low oxygen concentration. This damage is inhibited by scavengers of the hydroxyl radical, iron chelators and the specific proteins catalase and superoxide dismutase. The reactive radical species appears to arise from a Fenton‐type sequence in which iron is reduced by the mitomycin C semiquinone radical.
Carbon tetrachloride‐induced lipid peroxidation dependent on an ethanol‐inducible form of rabbit liver microsomal cytochrome P‐450
Authors: Johansson Inger , Ingelman-Sundberg Magnus
Published: July 2002

Treatment of rats with ethanol or rabbits with either imidazole or pyrazole, agents known to induce the ethanol‐inducible form of liver microsomal cytochrome P‐450 (P‐450 LMeb), caused, compared to controls, 3–25‐fold enhanced rates of CCl4‐dependent lipid peroxidation or chloroform production in isolated liver microsomes. No significant differences were seen when the rate of CCl4‐dependent lipid peroxidation was expressed relative to the amount of P‐450 LMeb in the various types of microsomal preparations. In reconstituted membranous systems, this type of P‐450 was a 100‐fold more effective catalyst of CCl4 metabolism than either of the cytochromes P‐450 LM2 or P‐450 LM4. It is proposed that the induction of this isozyme provides the explanation on a molecular level for the synergism seen of ethanol on CCl4‐dependent hepatotoxicity.
A new vitamin E analogue more active than α‐tocopherol in the rat curative myopathy bioassay
Authors: Ingold K.U. , Burton G.W. , Foster D.O. , Zuker M. , Hughes L. , Lacelle S. , Lusztyk E. , Slaby M.
Published: July 2002

Vitamin E owes its biological effects to its antioxidant activity. Kinetic and mechanistic studies on phenolic antioxidants in vitro have led us to design and synthesize all‐rac‐2,4,6,7‐tetramethyl‐2‐(4',8',12'‐trimethyltridecyl)‐5‐hydroxy‐3,4‐dihydrobenzofuran, 3. In the rat curative myopathy bioassay the acetate of this compound has 1.5–1.9 times the bioactivity of all‐rac‐α‐tocopherol acetate. This represents the first time that a rationally designed synthetic ‘vitamin’ has been found to have more activity in vivo than the corresponding natural vitamin.
Secondary structure prediction of human salivary proline‐rich proteins
Authors: Cid Hilda , Vargas Veronica , Bunster Marta , Bustos Sergio
Published: July 2002

Conformations associated with secondary structure in human salivary proline‐rich proteins A (PRPA), C (PRPC), P‐D and P‐E were predicted by analysis of their respective hydrophobicity profiles by computer programming. Structurally, PRPA and PRPC would present a globular head and a tail that consists of type 310 polyproline helices. P‐D and P‐E would be fibrilar molecules with helical zones of the polyproline 310 type. Alternatively for PRPA and PRPC, the head and tail would form one globular domain with the tail folding upon itself at places where random coils occur.
Repeating sequence homologies in the p36 target protein of retroviral protein kinases and lipocortin the p37 inhibitor of phospholipase A2
Authors: Weber Klaus , Johnsson Nils
Published: July 2002

Ca2+

(Intestine)
Lipid
Membrane protein
Phospholipase
Tyrosine phosphate
Single‐channel activity in sea urchin sperm revealed by the patch‐clamp technique
Authors: Guerrero A. , Sánchez J.A. , Darszon A.
Published: July 2002

Ionic fluxes are deeply involved in the response of spermatozoa to the egg. Using the patch‐clamp technique, we show for the first time single ion channel activity in sea urchin spermatozoa and spermatozoa heads. Due to their small size gigaseals were obtained in suspended cells by applying suction through the pipette. The rate of gigaseal formation was very low and improved to 6% (n = 1145) when flagella were detached from sperm. Current‐voltage curves created from single‐channel events showed conductances of approx. 65 and 170 pS, suggesting the presence of two types of channels. At least one appears to be a K+ channel.
Construction of a series of several self‐cleaving RNA duplexes using synthetic 21‐mers
Authors: Koizumi Makoto , Iwai Shigenori , Ohtsuka Eiko
Published: July 2002

Two fragments (21‐mers) containing consensus sequences for the self‐cleavage domain in transcripts of satellite DNA of the newt were chemically synthesized and found to be cleaved in the presence of Mg2+. The cleaved product contained the 3′‐terminal 2′,3′‐cyclic phosphate. Twenty‐five combinations of partially double‐stranded 21‐mer RNA which contained different bases within the consensus sequences and at the cleavage sites were tested for self‐cleavage. It seemed that guanosine 3′‐phosphate was not susceptible to transesterification at the cleavage site.
The tissue dependent expression of hamster P‐glycoprotein genes
Authors: Baas Frank , Borst Piet
Published: July 2002

Using a sensitive RNase protection assay, we have measured the levels of P‐glycoprotein mRNA in fourteen different Chinese hamster tissues and in three cell lines to determine whether a relationship exists between the level of P‐glycoprotein expression and the occurrence of primary and acquired multidrug resistance (MDR) in cancer. P‐Glycoprotein mRNA was detected in all tissues, suggesting that the protein is a normal constituent of the cell. High levels of P‐glycoprotein mRNA were found in oesophagus, testis and uterus. Intermediate levels in brain, lung and ovary, and a very low level in the adrenal gland, bladder, bone marrow, heart, kidney, liver and spleen. There is no obvious correlation between this expression pattern and the occurrence of primary or acquired MDR.
The prostate‐specific antigen gene and the human glandular kallikrein‐1 gene are tandemly located on chromosome 19
Authors: Riegman P.H.J. , Vlietstra R.J. , Klaassen P. , van der Korput J.A.G.M. , van Kessel A.Geurts , Romijn J.C. , Trapman J.
Published: July 2002

Using a prostate‐specific antigen cDNA as a hybridization probe, clones containing the kallikrein genes encoding prostate‐specific antigen, human glandular kallikrein‐1 and pancreas/kidney kallikrein were isolated from a human genomic library. Clones containing the prostate‐specific antigen gene and the human glandular kallikrein‐1 gene overlap and span a region of about 36 kb. The two genes are aligned in a head to tail orientation at a mutual distance of 12 kb. Southern blot analysis of DNA from a panel of human‐hamster hybrid cells with specific probes revealed the genes to be situated on chromosome 19. Assuming that the pancreas/kidney kallikrein gene is located in the same cluster, the distance to the prostate‐specific antigen gene and the human glandular kallikrein gene must be at least 15 kb.
Biosynthesis of cyclic 2,3‐diphosphoglycerate
Authors: Lehmacher Anselm , Vogt Anne-Barbara , Hensel Reinhard
Published: July 2002

Starting from 2‐phosphoglycerate the biosynthesis of cDPG comprises two steps: (i) the phosphorylation of 2‐phosphoglycerate to 2,3‐diphosphoglycerate and (ii) the intramolecular cyclization to cyclic 2,3‐diphosphoglycerate. The involved enzymes, 2‐phosphoglycerate kinase and cyclic 2,3diphosphoglycerate synthetase, were purified form Methanothermus fervidus. Their molecular and catalytic properties were characterized.
A novel approach to prediction of the 3‐dimensional structures of protein backbones by neural networks
Authors: Bohr H. , Bohr J. , Brunak S. , J. Cotterill R.M , Fredholm H. , Lautrup B. , Petersen S.B.
Published: July 2002

Three‐dimensional structures of protein backbones have been predicted using neural networks. A feed forward neural network was trained on a class of functionally, but not structurally, homologous proteins, using backpropagation learning. The network generated tertiary structure information in the form of binary distance constraints for the Cα
atoms in the protein backbone. The binary distance between two Cα
atoms was 0 if the distance between them was less than a certain threshold distance, and 1 otherwise. The distance constraints predicted by the trained neural network were utilized to generate a folded conformation of the protein backbone, using a steepest descent minimization approach.
Molecular cloning and sequence determination of four different cDNA species coding for α‐subunits of G proteins from Xenopus laevis oocytes
Authors: Olate Juan , Martinez Sixta , Purcell Patricia , Jorquera Hugo , Codina Juan , Bimbaumer Lutz , Allende Jorge E.
Published: July 2002

A cDNA library preprared from Xenopus laevis oocytes in λgt10 was screened with a mixture of three oligonucleotide probes designed to detect sequences found in different mammalian genes coding for a‐subunits of G‐proteins. In addition to a clone coding for a Gαo‐type subunit previously reported [(1989) FEBS Lett. 244, 188‐192] four additional clones have been found coding for different Gα protein subunits. By comparison with mammalian α‐subunits, these oocyte cDNAs correspond to two closely related Gas‐la, to a Gαi‐1 and to a Gαi‐3 species. The derived amino acid sequences showed that both Gαs species contain 379 residues, corresponding to the short species without the serine residue and with a calculated M
r of 42720. The Gαi‐1 gene encodes a 354 amino acid protein with an M
r, of 39000 and the Gαi‐3 encodes an incomplete open reading frame of 345 residues, lacking the first 9 amino acid residues at the NH2, terminus. All these Gα‐subunits showed high identity with their respective mammalian counterparts (75–80%), indicating a great degree of conservation through the evolution and the important cellular regulatory function that they play.
Selection of prosomes and prosomal RNA by immobilized viral RNAs
Authors: Horsch Andrea , Köhler Kurt , Ellwart-Tschürz Marlies , Schmid Hans-Peter
Published: July 2002

Viral messengers were used to select and purify prosomes and prosomal RNA from subribosomal fractions of HeLa cells and mouse erythroblasts. Adenovirus mRNA immobilized on oligo(dT)‐cellulose and tobacco mosaic virus RNA (TMV) sedimenting in sucrose gradients associated strongly with prosomes at high salt conditions forming intermolecular RNA‐RNA hybrids between prosomal RNA and viral RNA. Hybrid selection of small cytoplasmic RNAs with immobilized TMV‐RNA revealed a RNA species migrating at the same position as prosomal RNA. The possible existence of a box‐like sequence involved in hybridization will be discussed.
Sequence conservation in the α and β subunits of pyruvate dehydrogenase and its similarity to branched‐chain α‐keto acid dehydrogenase
Authors: Wexler Isaiah D. , Hemalatha Sullia G. , Patel Mulchand S.
Published: July 2002

Amino acid sequence comparison of 8α and 6β subunits of the α‐keto acid dehydrogenase (E1) component of the pyruvate dehydrogenase complex and branched‐chain α‐keto acid dehydrogenase complex from multiple species was performed by computer analysis. In addition to 2 previously recognized regions of homology in the α subunit, a 3rd region of extensive homology was identified in E1α, and may be one of the sites involved in subunit interaction. E1β contains 4 regions of extensive homology. Region 1 contains 10 amino acids that are homologous to a 10‐amino acid stretch in Escherichia coli E1. Regions 2 and 3 have sequence homologies with other dehydrogenases suggesting that these regions may be involved in catalysis.
Induction of heat‐shock proteins by glutamine The ‘feeding effect’
Authors: Cai Jie-Wei , Hughes Christine S. , Shen Jun-Wen , Subjeck John R.
Published: July 2002

Subconfluent, log‐phase Chinese hamster ovary cells induced the major heat‐shock proteins (hsp) when cells were refed, 40 hours after seeding. This method of inducing heat‐shock proteins was also obtained by refeeding with fresh serum‐free media, but not with media with a long shelf life or with media prepared without glutamine. It was observed that addition of glutamine alone to cultures at 40 hours post‐seeding induced heat‐shock proteins. Addition of ammonium chloride, however, had no discernible effect on heat‐shock protein synthesis. Northern blot analysis indicated that this phenomenon reflected an increase in the levels of message for the constitutive/inducible member of the hsp 70 family, but not the non‐constitutive member. To determine the effect of this induction on heat sensitivity, unfed and ‘heat‐shock‐induced’ refed cultures were heated at 45°C. No significant difference in cell survival was observed. Therefore glutamine is the necessary ingredient required for the induction of heat‐shock proteins and this method of inducing heat‐shock proteins does not alter heat sensitivity.
A new cyclopyrophosphate as a bacterial antistressor?
Authors: Ostrovsky Dmitry , Shipanova Irina , Sibeldina Lily , Shashkov Alexandr , Kharatian Elena , Malyarova Irina , Tantsyrev Georgy
Published: July 2002

In a number of bacteria an unusual glycosyl pyrophosphate (31P NMR signal chemical shift at about −15 ppm) was detected when the cells were subjected to oxidative stress. This substance from Brevibacterium ammoniagenes has now been identified as 2‐methyl‐butan‐1,2,3,4‐tetraol‐2,4‐cyclopyrophosphate, which is accumulated in the cell under certain conditions in concentrations of about 50 mM. It is now suggested that this compound is the long sought after bacterial antistressor.
Ordered phosphorylation of a duplicated minimal recognition motif for cAMP‐dependent protein kinase present in cardiac troponin I
Authors: Mittmann K. , Jaquet K. , Heilmeyer L.M.G.
Published: July 2002

Cardiac troponin I contains two adjacent serines in sequence after three arginine residues thus making up a minimally duplicated recognition motif for cAMP‐dependent protein kinase. In a synthetic peptide, PVRRRSSANY, the two serine residues are phosphorylated sequentially with the intermediate formation of a monophosphorylated species according to the following reaction sequence: Peptide k
1|→ Peptide‐P k
2|→ Peptide‐P2. The calculated rate constants are: k
1 = 0.435 min−1 and k
2 = 0.034 min−1. Sequence analyses of the monophosphopeptide and its tryptic fragments show that the predominant monophosphoform carries phosphate at the second serine.
Parathyroid hormone‐related peptide can regulate the growth of human lung cancer cells, and may form part of an autocrine TGF‐α loop
Authors: Burton Paul B.J. , Knight Derek E.
Published: July 2002

Parathyroid hormone‐related peptide (PTHrP) and transforming growth factor‐α (TGF‐α) were found to stimulate proliferation of human lung cancer cells (BEN‐57). TGF‐α stimulated PTHrP secretion from these cells. The polyclonal antisera raised against PTHrP significantly inhibited the growth of BEN‐57 cells, and also the proliferation induced by TGF‐α. Treatment of cells for up to 10 days with either a PTHrP receptor antagonist (PTHrP(7–34)) or PTHrP antiserum significantly inhibited the subsequent growth of these cells. We suggest that PTHrl may be a component of a complex autocrine loop involving TGF‐α.
Effect of pancreatic type phospholipase A2 on isolated porcine cerebral arteries via its specific binding sites
Authors: Nakajima Masatoshi , Hanasaki Kohji , Ueda Motohiko , Arita Hitoshi
Published: July 2002

The addition of porcine pancreatic group I phospholipase A2 (PLA2‐I) produced a transient contraction followed by a relaxation in helical strips of porcine cerebral arteries. Its ED10 value (2.3 nM) was almost identical to the K
4 value(3.9 nM) calculated from the specific binding of 125I‐labeled porcine PLA2‐I in cultured porcine cerebral arterial smooth muscle cells. Type‐specific action of PLA2s and homologous desensitization strongly implicated the involvement of PLA2‐I‐specific sites in the response. The transient contraction was abolished by treatment with indomethacin as well as by the removal of endothelium, indicating the dependence of vasoconstrictor prostoglandins synthesized by PLA2‐I in endothelium. The PLA2‐I‐induced relaxation response was also observed in bovine and cat cerebral arteries, thus providing a new aspect of PLA2‐I as a vasoactive substance.
1H NMR study on the conformation of bacitracin A in aqueous solution
Authors: Kobayashi Naohiro , Takenouchi Takato , Endo Satoshi , Munekata Eisuke
Published: July 2002

The conformation of bacitracin A, a widely used cyclic dodecapeptide antibiotic in aqueous solution, has been investigated using 500 MHz 1H NMR and molecular modeling. Findings revealed that a region (residues 1–6) is folded over the cyclic ring, resulting in metal coordination sites, a thiazoline ring, and Glu4 and His10 being proximate to each other.
Induction of thromboxane synthase and prostaglandin endoperoxide synthase mRNAs in human erythroleukemia cells by phorbol ester
Authors: Ihara Hayato , Yokoyama Chieko , Miyata Atsuro , Kosaka Tetsuya , Nüsing Rolf , Ullrich Volker , Tanabe Tadashi
Published: July 2002

The effects of 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA) on the mRNA levels of two enzymes, thromboxane synthase (TXS) and prostaglandin endoperoxide synthase (PES), responsible for the synthesis of thromboxane A2 from arachidonic acid, were studied in human crythroleukemia (HEL) cells by RNA blot analysis. TPA induced both TXS and PES mRNAs in HEL cells in a dose‐dependent manner at 36 h. The half‐maximal and maximal effects for the induction of both mRNAs ware at approximately 3×10−9 M and at 10−8 M, respectively. TXS and PES mRNA levels increased in a time‐dependent fashion by TPA, and reached to 7‐ and 3.5‐fold of the control, respectively after 48 h of TPA treatment. These results suggest that expression of TXS and PES genes in HEL cells were simultaneously stimulated by TPA.
Comparison of ubiquinol and cytochrome c terminal oxidases
Authors: Musser Siegfried M. , Stowell Michael H.B. , Chan Sunney I.
Published: July 2002

There have been numerous instances in the recent literature where the properties of ubiquinol and cytochrome c terminal oxidases are compared. Here we specifically examine the cytochrome bo
3‐type ubiquinol oxidase from Escherichia coli and the cytochrome aa
3‐type cytochrome c oxidases. A second redox‐active copper site (CuA) is present only in the cytochrome c oxidases and the physiological electron donors for the two enzymes are different (ubiquinol‐8 vs. ferrocytochrome c). In our opinion, these differences are significant and most likely indicate that distinct turnover mechanisms are operative in the two enzymes.
Mass spectrometric evidence for a disulfide bond in aequorin regeneration
Authors: Ohmiya Yoshihiro , Kurono Sadamu , Ohashi Mamoru , Fagan Thomas F. , Tsuji Frederick I.
Published: July 2002

Tryptic digests of purified recombinant apoaequorin were analyzed, before and after reduction with DTT, by fast atom bombardment mass spectrometry. The results showed that apoaequorin contains a disulfide bond between Cys145 and Cys152 and that the reduction of this bond is involved in the regeneration of aequorin.
In vitro activation of pro‐cathepsin B by three serine proteinases: leucocyte elastase, cathepsin G, and the urokinase‐type plasminogen activator
Authors: Dalet-Fumeron Veronique , Guinec Nathalie , Pagano Maurice
Published: July 2002

In vitro activation of pro‐cathepsin B purified from ascitic fluid of ovarian carcinomas by serine proteinases was studied. Both elastase and cathepsin G from human leucocytes were found to be activators, on the basis of generation of cathepsin B activity and processing of the precursor. These results represent a new cooperative pathway between cancer cells and host cells. The urokinase‐type plasminogen activator activated pro‐cathepsin B faster than leucocyte proteinases. A new relationship is emerging between the cysteine proteinases and the plasmin‐activation system. Both pathways suggest an important role of cathepsin B in the proteolytic cascade associated with tumour invasion.
Transcriptional regulation of the isocitrate lyase encoding gene in Saccharomyces cerevisiae
Authors: Fernandez Ernestina , Fernandez Maria , Moreno Fernando , Rodicio Rosaura
Published: July 2002

In this work, we studied the transcriptional regulation of isocitrate lyase synthesis. In Northern blot analyses we first showed that the steady‐state ICL1 mRNA levels depend on the carbon source used for growth. In addition, we determined the kinetics of transcriptional repression upon a shift of ethanol‐grown cells to glucose and of the induction when cells were transferred from glucose to ethanol. By deletion analyses as well as by studying the influence on expression of different fragments cloned into the heterologous CYC1 promoter lacking its own UAS sequences, we defined UAS and URS elements in the ICL1 promoter. A region mediating the control by CAT3, a gene also involved in the control of expression of other genes subject to carbon catabolite repression, was found to overlap with one of these UAS elements.
A synthetic model of collagen structure taken from bovine macrophage scavenger receptor
Authors: Tanaka Toshiki , Wada Yayoi , Nakamura Haruki , Doi Takefumi , Imanishi Takeshi , Kodama Tatsuhiko
Published: July 2002

A putative collagen structure from macrophage scavenger receptors binds to a wide range of ligands. In order to elucidate the ligand's binding mode, this collagen structure was constructed using short peptides. This was accomplished by the reaction of a tri‐bromoacetylated branched peptide with a purified unprotected 25‐residue peptide, which contained Cys, 4 repeats of the triplet, Gly‐Pro‐Hyp, and 12 residues from the bovine macrophage scavenger receptor (residues 332 to 343). The three identical 25‐residue peptides are linked at the N‐terminus. CD and NMR spectra of the N‐terminus cross‐linked tripeptide show that it forms a collagen structure below 10°C and an extended structure at high temperature with a midpoint of 20° C.
The functional effects of mutations Thr673→ Asp and Ser702→ Asp at the Pro‐directed kinase phosophorylation sites in the C‐terminus of chicken gizzard caldesmon
Authors: Redwood Charles S. , Marston Steven B. , Gusev Nikolai B.
Published: July 2002

We expressed the C‐terminal 99 amino acids of chicken gizzard caldesmon (658C) and two point mutants in which the preferred phosphorylation sites of MAP kinase and p34cdc2kinase, Ser702and Thr673were altered to aspartic acid. The T673D mutant was indistinguishable from 658C but S702D was not phosphorylated by MAP kinase, was significantly less potent as an inhibitor of actin‐tropomyosin activation of myosin MgATPase, and bound less actin‐tropomyosin at low concentrations. Thus Ser702is involved in the tropomyosin‐dependent inhibitory mechanism of caldesmon, and its phosphorylation by MAP kinase or p34cdc2kinase could modulate caldesmon function.
In vitro expression and activity of lycopene cyclase and β‐carotene hydroxylase from Erwinia herbicola
Authors: Hundle Bhupinder S. , O'Brien David A. , Beyer Peter , Kiemig Hans , Hearst John E.
Published: July 2002

The cyclisation of lycopene to β‐carotene and the hydroxylation of β‐carotene to zeaxanthin are common enzymatic steps in the biosynthesis of carotenoids in a wide range of bacteria, fungi, and plants. We have individually expressed in E. coli the two genes coding for these enzymatic steps in Erwinia herbicola. The cyclase and hydroxylase enzymes have apparent molecular weights of 43 kDa and 22 kDa, respectively, as determined by SDS‐PAGE. Hydroxylase in vitro activity was obtained only in the cytoplasmic fraction. Cyclase also demonstrated enzyme activity in a crude cell‐free lysate, although to a lesser extent.
The N‐ethylmaleimide‐sensitive fusion protein (NSF) is preferentially expressed in the nervous system
Authors: Püschel Andreas W. , O'Connor Vincent , Betz Heinrich
Published: July 2002

NSF and SNAPs (soluble NSF attachment proteins), originally identified as cytosolic components of intracellular vesicular transport mechanisms, have recently been implicated in Ca2+‐triggered neurotransmitter release from synaptic terminals. Here, we have investigated the temporal and spatial expression pattern of the rodent NSF and SNAP genes. A single transcript of 4.5 kb is highly expressed in rat brain, whereas only minor amounts of NSF mRNA are found in liver, kidney, heart, lung and skeletal muscle. In situ hybridisation revealed NSF transcripts as early as embryonic day 10 preferentially in the nervous system of mouse embryos. In the adult brain NSF is widely expressed with particularly high levels in the hippocampus. An identical expression profile was observed for α/β‐SNAP. Our data are consistent with a central function of NSF and SNAPs in neurotransmission.