bluuebunny/crossref_metadata_2025 huggingface dataset:
Features of Rehabilitation Therapeutic Treatment of Patients who have come through Coronavirus Disease
Authors: Gimbatova Aishat Iskhakovna , Gabrielyan David Avakovich , Timokhina Tatiana , Kostikov Andrey Petrovich , Shekhovtsova Anastasiia Viktorovna , Sokolova Aleksandra Alekseevna
Published: June 2021

The purpose of the article is to consider the features of restorative therapeutic treatment of patients who have suffered a coronavirus infection. This disease can cause respiratory, physical and psychological dysfunction in patients. Therefore, restorative therapeutic treatment and rehabilitation are critical for both hospitalized and discharged patients with COVID-19.

The author believes that many patients who have a history of a previous coronavirus infection note that they experience various unpleasant symptoms even six months after receiving negative tests. So, such patients may experience weakness, shortness of breath, tachycardia, instability of mental states, and so on. At the same time, against the background of an active fight against the spread of COVID-19, measures related to the rehabilitation of convalescents are receding into the background, which is fundamentally wrong. The paper analyzes the literature devoted to the problem of rehabilitation treatment after a coronavirus infection, and also applies comparative and comparative research methods.

The author concludes that complications after COVID-19 can significantly worsen the quality of life of patients. For this reason, specialists need to identify the degree and level of complications and patients who have suffered a coronavirus infection as early as possible in order to prescribe appropriate medication or physiotherapy.
Effectiveness of Standard post-operative Care on Post-operative Outcome Evaluation among Women Undergone Lower Segment Cesarean Section in Selected Hospital: A Narrative Review
Authors: Kumari Prabha , Singh Maharaj , Sunita .
Published: June 2021

Background: Present study focuses on effectiveness of standard post-operative care on post-operative evaluation among women undergone lower segment cesarean section in selected hospitals. Standard post operative care basically stands for list of non-pharmacological or pharmacological measures, procedures and methods adopted by the investigator/care giver during post-operative period of women undergone lower segment cesarean section for the reduction of negative post-operative outcomes or complications and to fasten recovery. Present study encompasses articles on post-operative care in terms of one, two or more than two or three measures or interventions as post-operative care measures adapted to reduce post-operative complications like use of music therapy, foot-hand massage therapy, planned early ambulation, planned ambulation techniques etc. for post-operative complications and pain relief.

Objective: To identify various post-operative measures or care which can be included in standard post-operative care list and its effectiveness on post-operative outcome evaluation among women undergone Lower segment cesarean section in selected hospitals.

Methods: Articles were searched in Google scholar, Pub-med, Medline and research gate by using keywords post-operative care of cesarean section, post-operative outcomes of cesarean section and Cesarean mother.

Result: Review includes seven articles showing different non-pharmacological or pharmacological measures or techniques to overcome post-operative complications for early recovery and better outcomes. These measures can be included in standard post-operative care list for positive post-operative maternal and fetal outcomes. These measures include early planned ambulation, planned ambulation techniques, music therapy, Foot and hand massage therapy and use of certain analgesia for pain management in post cesarean women if needed, as per the patient’s condition.

Conclusion: Conclusion of the review revealed that there are various measures which can be standardized in post-operative outcome evaluation among women undergone lower segment cesarean section.
An Overview on the Biosynthetic Pathways and Medicinal Values of Secondary Metabolites
Authors: Armarkar Ashwini V. , Mahure Divyani S. , Pounikar Anita R. , Bhagat Rani T. , Nagare Dhanashri N.
Published: June 2021

Nature is a treasure hunt of novel molecules which are extensively used by humans for their medicinal values for millennia. Herbal medicines have emerged as life savers all over the world due to their multifunctional and diverse array of activities on the human body. Plants supply numerous bioactive molecules which are capable of treating disease conditions. Plants produce varieties of phytochemicals which are commonly recognized as primary and secondary metabolites. Secondary metabolites are inimitable resources for exploitation for pharmaceuticals, food additives e.t.c. They are often involved in the control of abiotic or biotic stress. They play distinctive roles in the defense mechanism of plants, act as insect repellant, regulate signaling pathways, and also exhibit varied range of pharmacological actions such as antibacterial, antioxidant, anticancer anti-diabetic and others. Secondary metabolites are derived from one or mixed pathways that give rise to formation of various compounds i.e., alkaloids, volatile oils, tannins, glycosides and resins etc. which have great importance in modern medicine. Isopentenyl diphosphate and Shikimic acid pathway serve as primary precursors for the synthesis some of types of secondary metabolites. The present review deals with the brief introduction, significance of secondary metabolites in the plants, biosynthetic pathways, and therapeutic importance of some commonly known secondary metabolites.
Prevalence of Urinary Tract Infection and Drug Resistance among Infants and Children in Pakistan
Authors: Sohail Muhammad , Shahid Maisam Ali , Sajjad Kumail , Ayub Muhammad , Akram Qasir , Alyas Sobia , Javed Yousma , Rana Muhammad Asim , Ali Qurban , Hafeez Muhammad Mansoor
Published: June 2021

In infancy most common and a serious disease is known as Urinary tract infection (UTI). The aim of the current study is to evaluate the frequency and antibiotic resistant among infant presenting with UTI in Lahore General hospital, Lahore-Pakistan. The current observational study comprised 100 (n=100) cases diagnosed with UTI. The study was conducted at Lahore General Hospital, Pakistan, between March 2019 and January 2020. Data were collected and recorded on specified proformas that had information on demographics, biochemical analysis, and complaints, as well as the antibiotic used and blood cultures. The most common gram-negative bacilli (GNB) causing UTI was E. coli constitute about 72% followed by Klebsiella (11%), Enterobacter (7%) and Enterococcus (4%). Most of the isolated pathogens are highly resistant to ampicillin, cotrimoxazole, and cephalexin (72%–95%), have intermediate sensitivity to third- generation cephalosporins, and high sensitivity to amikacin, nitrofurantoin and ciprofloxacin. Similarly, the other pathogens showed different degree of occurrence and resistance against empirically used antibiotics. The most prevalent UTI causative organism among children was E. coli. The insulated microorganisms were highly resistant to ampicillin, and highly sensitive to amikacin.
A Prospective Observational Study to Find out Prevalence of Polypharmacy, Co-Morbidities and Drug-Related Problems among Type 2 Diabetes Patients at a Tertiary Care Teaching Hospital
Authors: Hadia Rajesh , Thakkar Mihir , Shah Shivani , Patel Prati , Gohel Kushalkumar , Joshi Dhaval , Rathod Trupal , Lakhani Jitendra , Rajput Hemraj Singh , Maheshwari Rajesh
Published: June 2021

Background: Presently India, depicts 49% of the world’s diabetes burden, with an approximated 72 million cases in 2017. Comorbidities make type 2 Diabetes Mellitus (DM) management complicated for health care providers. Therefore, patients have been prescribed multiple drugs. As a result of polypharmacy, the chances of the occurrence of drug-related problems (DRPs) increase.

Objective: The present study aimed to find out the prevalence of comorbidities, polypharmacy, and drug-related problems among type 2 DM patients.

Methodology: A prospective observational study was conducted for six months among 110 patients having type 2 DM. Based on inclusion and exclusion criteria data were collected. All collected cases were analysed for the number of medications prescribed and comorbidities present. Prescriptions were further assessed to find out the drug-related problem.

Results: Comparing the gender proportionality, male represents 59% (n=65) and female represents 41% (n=45) of the study population. The present study evaluated that 91% (n=100) of patients were having at least one comorbid condition or complication along with diabetes, the most common being cardiovascular disorders. The average number of drugs prescribed is 7.86±7.83 per patient. Polypharmacy was observed in 84.54% (n=93) of the patients. A total of 180 DRPs were found among 110 patients.

Conclusion: The present study concluded that the increase in the number of co-morbidities significantly increases the burden of polypharmacy which can further lead to DRPs. Reducing the number of pills and the presence of a clinical pharmacist in prescription monitoring can help to alleviate the problems of polypharmacy and its consequences.
Modification in the Oncology Field during COVID-19 Pandemic
Authors: Turkistani Fatema , Sawad Aseel Bin
Published: June 2021

Background: COVID-19 outbreak impacted all healthcare specializations including the oncology field.

Objective: To discuss in detail the varied effects of COVID-19 on the oncology field.

Methods: For this narrative review, the researchers relied only on accredited and peer-reviewed resources. As such, the references of this paper were mostly taken from Google Scholar and online repositories, such as PubMed, which hosts the National Center for Biotechnology Information (NCBI), Science Direct, JSTOR, and The Lancet.

Results: COVID-19 and the necessity of physical distancing have significantly changed the practice in the oncology field. The use of telehealth is widely adopted, physicians are recommended to consider other forms of treatment, and decisions on immediate cancer treatment depend on the level of risk of progression with cancer care delay. Cancer treatment delay is causing the highest mortality rate for patients who are suffering from cancer of the bladder, lungs, ovary, stomach, and esophagus. COVID-19 has shifted the focus of healthcare professionals away from other life-threatening diseases, like cancer. In the United Kingdom, it has been projected that there is at least a 20% increase in the expected cancer-related mortality rate. This is due to abrupt changes in diagnosing and treating cancer patients, physical distancing protocols, economic downfall, as well as the public's behavior in opting for medical assistance.

Conclusion: With the spread of COVID-19, the situation has become more difficult for cancer patients. The mortality rate for cancer patients has worsened during COVID-19. Health professionals working in the oncology field are also devastated by COVID-19.
Model tests for tilting control of suction bucket foundation for offshore wind turbine with path points
Authors: Kim You-Seok , Lee Jong-Pil
Published: June 2021

In offshore wind farms, tilting control based on a single-basket suction bucket foundation is a significant problem. In a single-basket suction bucket foundation, the tilting control of the foundation is possible by arranging the cells inside and controlling the pressure of each cell. However, the pressure of each cell must be finely controlled. Studies on this topic have been conducted, but no specific tilting control method has been developed. This paper presents experimental model results for tilting control obtained during the installation of a suction bucket foundation consisting of three internal cells. Tilting control was performed by independently controlling the internal pressure of each cell. A 1:100 scale model was used, and the ground condition was sandy. Four cases of tilting control tests for the model foundation were used with multiple combinations of internal positive, negative, or both pressures of each cell. It was found that the tilting control was within 5° during the installation and operation stages. There was a tilting control limit for operation based on the model with only negative pressure; therefore, 5° tilting control was achieved by combining the positive pressure.
A study on the vertical bearing capacity according to the inclination angle of single batter piles and the relative density of sand under vertical loads
Authors: Yun Seong-Kyu , Kim Jiseong , Kang Minsu , Kang Gichun
Published: June 2021

This study analyzed the increase in the compressive bearing capacity of single-sloped piles according to the relative density and inclination angle when the batter piles are subjected to a vertical load in non-adhesive sand. An experiment was conducted with inclination angles of 0, 10, 20, 30, and 40, and the relative density of sandy soil was divided into 31% (loose), 53% (medium), and 72% (dense) and compared with the results of the earlier research. As a result of the experiment, when the relative density of the ground was medium and dense, the bearing capacity was greater than that of the vertical pile (0°) at all angles. The tendency noted was that the bearing capacity of the pile increased from vertical to 20° and gradually decreased after 20°. The same tendency was also exhibited by loose sandy soil, but with less bearing capacity than the vertical pile (0°) except for 20°.
Seismic Performance Evaluation of Agricultural Reservoir Embankment through Shaking Table Tests
Authors: Lee Younghak , Ryu Junghyun , Yoon Bora , Heo Joon , Lee Dalwon
Published: June 2021

In this study, shaking table tests were performed to compare and analyze the acceleration response, displacement behavior, and pore-water pressure behavior of reservoirs with parapets installed to prevent overtopping of deteriorative homogeneous reservoirs. During the shaking table tests, the experimental conditions were divided into four cases considering the range and magnitude of seismic acceleration according to national standards. The vibration-type waveform (Gongen) and shock-type waveform (Minogawa) were applied as input waveforms. The acceleration amplification ratios of both vibration- and shock-wave types were the largest in the dam crest, and the amplification ratio decreased as the design earthquake acceleration increased. In addition, the horizontal displacement was maximum on the upstream slope, owing to the influence of seepage water, and the vertical displacement was maximum on the dam crest, owing to the self-weight effect of the parapet structure. A comparison of the waveform results indicates that the vibration-type waveform may exhibit a more significant effect on the embankment zone displacement than the shock-type waveform. However, when the safety standards for the horizontal displacement, settlement ratio, and excess pore-water pressure ratio were applied, the embankment was stable within the allowable range in both the shock-type and vibration-type waveforms. Therefore, the parapet structure is expected to influence the overflow resistance and stability of embankments positively.
A Study on the Improvement of the Disaster Relief Donation System
Authors: Sung Gihwan , Kim Yongsang , Yoo Sunwoong
Published: June 2021

This study discussed the need for restructuring the disaster relief fundraising system in South Korea. To that end, it examined the management system and characteristics of disaster relief donations. In addition, the study drew implications by comparing domestic and overseas disaster relief donation systems and analyzing important precedents related to disaster relief donations. The analysis revealed that the domestic disaster relief fundraising system is based on the classification of disasters into natural and social disasters, which the study contended is not suitable in the current scenario given the recent trend of large and complex disasters. Overseas disaster donation systems were not found to be based on such a dual classification. Moreover, it was confirmed that various forms of support were being provided to revitalize donations overseas. It may also be unconstitutional for donations to be based on such a classification because the system violates the basic spirit of the Constitution by limiting the right to freedom and pursuit of happiness of donors and charitable organizations participating in disaster relief funding. Historical changes in the donation law have changed the orientation from regulation to promotion of a donation system with a mature donation culture. In this context, the following proposals were made to improve the domestic disaster relief donation system. First, the study recommended the unification of the donation system for natural and social disasters in light of the occurrence of multiple disasters. Second, it advocated a transition away from the current system of monopolistic fund management in order to revitalize various disaster relief fundraising institutions.
Analysis of Patent Trends for Fire Protection Technology based on IoT and ICT Technology
Authors: Ryu Eunmi , Yoon Hyeonggu , Yeo Inhwan , Cho Kyungsuk
Published: June 2021

Technology for the 4th Industrial Revolution, including ICT, IoT, and big data, has been combined with various other technologies. As an example, these technologies combined with fire protection technology can help protect lives and minimize property damage due to fires. In this study, the patents for fire protection technology using IoT and ICT technologies were investigated from 1999 to 2018 in Korea, the United States, Japan, and Europe. The technology patents were analyzed for international patent status and technological competitiveness by country. Although there was a slow increase in the overall number of patents for the fire protection technology from 1999 to 2014, the number has increased rapidly since 2015. The analysis in this study was intended to afford basic data on existing research to understand the trend and impact of the incorporation of the technology for the 4th Industrial Revolution into extant technology.
Solubilized DNA‐dependent RNA polymerase from human placenta: A Mn2+‐dependent enzyme
Authors: Voigt H.-P. , Kaufmann R. , Matthaei H.
Published: July 2002

The 100‐fold purified RNA polymerase activity from human placenta is completely dependent upon added DNA. The enzyme is most active at 3 mM Mn2+ in the presence of 100 mM (NH4)2SO4. Denatured DNA is a better template than native DNA. α‐Amanitin completely inhibits the incorporation of 3H‐UMP, while rifampicin has no influence upon the enzymatic activity.
The observation of quasi‐molecular ions from a tiger snake venom component (Mr 13 309) using 252Cf‐plasma desorption mass spectrometry
Authors: Kamensky I. , Håkansson P. , Kjellberg J. , Sundqvist B. , Fohlman J. , Peterson P.A.
Published: July 2002

A method involving fast heavy‐ion bombardment of a solid sample called 252Cf‐plasma desorption mass spectrometry has been used to study of a non‐enzymatic, non‐toxic phospholipase homolog from Australian tiger snake (Notechis scutatus) venom. The protein consists of 119 amino acids in a single polypeptide chain cross‐linked by 7 disulfide bridges. The isotopically averaged molecular mass as determined by protein sequence analysis is 13 309 atomic mass units (amu). The mass distributions were studied by means of time‐of‐flight measurements. Quasi‐molecular ions associated to the molecule and its dimer were observed. The mass of the quasi‐molecular ion corresponding to the molecule was determined to be 13 285 ± 25 amu.
Interaction of polymerases with 2′‐deoxyuridine‐5′‐triphosphate spin‐labeled at the 5‐position
Authors: Warwick-Koochaki P.E. , Hakan A. , Bobst A.M.
Published: July 2002

2′‐Deoxyuridine‐5′‐triphosphate spin‐labeled at the 5‐position with N‐[1‐oxyl‐2,2,6,6‐tetramethyl‐4‐piperidinyl]‐O‐ was found to be an inhibitor of some DNA and RNA polymerases including avian myeloblastosis virus reverse transcriptase. Furthermore, the spin‐labeled nucleotide was found to be incorporated internally into polydeoxythymidylic acid via reverse transcriptase to an extent of 1.0 spin‐labeled base per 103 bases. The incorporation, monitored by electron spin resonance, is analogous to some other nucleotide inhibitors of polymerases, and the results indicate that it may be feasible to obtain sequence specific, spin‐labeled DNA, enzymatically.
Changes induced by hydrazine in optical spectra of cytochrome oxidase
Authors: Markossian Kira A. , Paitian Norair A. , Nalbandyan Robert M.
Published: July 2002

Under aerobic conditions hydrazine reduces haem a of cardiac cytochrome oxidase and brings about the formation in optical spectra of a new band at 845 nm, whereas under anaerobic conditions hydrazine reduces both haems, a and a
3, as well as EPR‐detectable copper, CuA, and results in the band at 845 nm. The formation of this band was sensitive to inhibitors of oxygen binding. It is suggested that the band at 845 nm reflects changes induced by hydrazine in CuB which in the resting enzyme is EPR‐undetectable.
Proton magnetic resonance studies of 7 Fe ferredoxins
Authors: Nagayama K. , Ohmori D. , Imai T. , Oshima T.
Published: July 2002

Two types of iron‐sulfur clusters, [3 Fe–3 S] and [4 Fe–4 S], were identified by 1H‐NMR in ferredoxins from Thermus thermophilus, Mycobacterium smegmatis and Pseudomonas ovalis. The [4 Fe–4 S] clusters always showed the redox couples which had potentials lower than that of the [3 Fe–3 S] clusters.
Carbocyanine dyes used as fluorescent triplet probes for measuring slow rotational diffusion of lipids in membranes
Authors: Johnson Pauline , Garland Peter B.
Published: July 2002

A method for measuring the slow rotational diffusion of lipids or lipid domains in membranes has been developed. It covers the time range from 20μs–5 ms, and has a sensitivity > 5 × 104 molecules of probe. The method uses acyl‐substituted carbocyanine dyes as fluorescent triplet probes and a laser‐microscope combination for excitation and measurement of the triplet state. Rotation rates in dimyristoylphosphatidylcholine vesicles were sensitive to the liquid‐to‐gel transition. Slow rotations with relaxation times of about 100μs were detected at the transition temperature region.
The ionic channel of the nicotinic acetylcholine receptor is unable to differentiate between the optical antipodes of perhydrohistrionicotoxin
Authors: Spivak C.E. , Maleque M.A. , Takahashi K. , Brossi A. , Albuquerque E.X.
Published: July 2002

The enantiomers of perhydrohistrionicotoxin were studied in their effects on endplate currents recorded at the junctional region of sartorius muscles of Rana pipiens. The two optical antipodes progressively decreased the peak amplitude of the endplate currents and were indistinguishable from each other at all times. The enantiomers shortened equally the time constants for endplate current decay, but did not alter their voltage sensitivities. Although perhydrohistrionicotoxin contains 4 chiral centers, complete steric inversion does not alter its effects on the acetylcholine receptor—ion channel complex. By contrast the recognition site of the AcChR is extremely sensitive to any change in the chirality of agonists.
Dynamic structure of metallothionein
Authors: Vašák Milan , Berger Christine , Kägi Jeremias H.R.
Published: July 2002

Molecular sieve chromatography of rabbit liver metallothionein at different electrolyte concentrations revealed that this protein undergoes an increase in Stokes radius from 1.50 to 1.78 nm when the ionic strength is lowered from 0.5 to 0.015 indicating a change in molecular shape and/or hydration. The variation in ionic strength also affects the far‐UV circular dichroism of metallothionein reflecting a conformational transition in the protein. The effects are attributed to changes in intramolecular repulsion between the strongly negatively charged metal‐thiolate clusters of the protein. It is suggested that metallothionein exists in at least two interchangeable conformational states which differ in hydrodynamic properties and whose equilibrium concentrations are determined by the electrostatic free energy of the system.
Does casomorphin have a functional role?
Authors: Petrilli Pasquale , Picone Delia , Caporale Carlo , Addeo Francesco , Auricchio Salvatore , Marino Gennaro
Published: July 2002

Degradation of buffalo β‐casein by various physiological enzymes was studied. Digestion with gastric and pancreatic proteases plus leucine aminopeptidase did not release casomorphins but a putative precursor (procasomorphin) which was further digested by brush border peptidases into peptides differing from casomorphins.
High‐pressure enzyme kinetics
Authors: Chernyak V.Ya. , Drachev V.A. , Nikolaeva N.G. , Chernyak B.V.
Published: July 2002

A newly designed optical cell allows an enzyme reaction to be started under high pressure and makes it possible to begin measurement of the reaction rate after a ‘dead time’ no longer than 1–2 s. This device was used to study the kinetics of lactate dehydrogenase reaction at 1 kbar. At this pressure lactate dehydrogenase from rabbit muscle exhibited a rapid deactivation in the presence of NADH if pyruvate was absent. After addition of pyruvate the reaction was initiated and proceeded at a constant rate, i.e., without loss of enzyme activity. It is suggested that pyruvate markedly increases the association constant of this tetrameric enzyme.
Two‐dimensional Fourier transform 1H NMR studies of ribosomal protein E‐L30
Authors: van de Ven F.J.M. , de Bruin S.H. , Hilbers C.W.
Published: July 2002

Two‐dimensional Fourier transform 1H NMR spectra of ribosomal protein L30 of Escherichia coli MRE 600 were recorded at 500 MHz both in H2O and in 2H2O. From the available data we infer (i) the existence of one or more hydrophobic domains in the molecule, (ii) at least two helical regions and (iii) the presence of an antiparallel β‐sheet involving two threonines.
Relationships between the effects of redox potential, α‐Thenoyltrifluoroacetone and malonate on O2− and H2O2 generation by submitochondrial particles in the presence of succinate and antimycin
Authors: Ksenzenko Marina , Konstantinov Alexander A. , Khomutov Gennadiy B. , Tikhonov Alexander N. , Ruuge E.K.
Published: July 2002

The rate of the antimycin‐induced H2O2 and O2
XXXgeneration in beef heart submitochondrial particles is maximal at the [succinate]/[fumarate] ratio of 1:5 and decays at both higher and lower redox potentials. Succinate dehydrogenase inhibitors, such as TTFA or malonate, stimulate active oxygen production in the presence of excess succinate but are inhibitory at E
h, values more positive than the optimal. The modulation of O2
xxx and H2O2 generation by these inhibitors can be explained by their effects on the steady‐state redox potential(s) of the component(s) of mitochondrial site 2.
Chymotrypsin modified with polyethylene glycol catalyzes peptide synthesis reaction in benzene
Authors: Matsushima Ayako , Okada Masato , Inada Yuji
Published: July 2002

Chymotrypsin was modified in the zymogen form with 2,4‐bis(O‐methoxypolyethylene glycol)‐6‐chloro‐s‐triazine (activated PEG2), followed by activation with trypsin. The modified enzyme was soluble in benzene and retained its enzymic activity. Acid‐amide bond formation by the modified enzyme proceeded efficiently in benzene: N‐benzoyltyrosine butylamide was made from N‐benzoyl‐L‐tyrosine ethyl ester and n‐butyl‐amine, and benzoyltyrosine(oligo)phenylalanine ethyl esters were formed from N‐benzoyl‐L‐tyrosine ethyl ester and L‐phenylalanine ethyl ester.
The rate constant for ATP hydrolysis by polymerized actin
Authors: Pollard Thomas D. , Weeds Alan G.
Published: July 2002

Polymerized actin hydrolyzes bound ATP in a reaction that depends on the concentration of polymerized ATP‐actin, not on the rate of incorporation of ATP‐actin into the polymer. The apparent first order rate constant is about 0.07 s−1 at 21°C in 50 mM KCl with 1 mM MgCl2 or CaCl2.
N‐terminal amino acid sequence of an insect neurohormone, melanization and reddish coloration hormone (MRCH): heterogeneity and sequence homology with human insulin‐like growth factor II
Authors: Matsumoto Shogo , Isogai Akira , Suzuki Akinori
Published: July 2002

Amino acid sequence
Insect neurohormone
Insulin‐like growth factor‐II
Sequence homology
Amino terminal heterogeneity
Cuticular melanization
Binding specificities of the lectins PNA, WGA and UEA I to polyvinylchloride‐adsorbed glycosphingolipids
Authors: Molin Kent , Fredman Pam , Svennerholm Lars
Published: July 2002

The binding specificities of the lectins PNA (peanut agglutinin), WGA (wheat germ agglutinin), and UEA I (Ulex europeus agglutinin I) against glycosphingolipids were investigated using an enzyme‐linked immunosorbent assay (ELISA), utilizing the biotin‐avidin system for detection of bound lectin. PNA showed the highest affinity to GA1, but also bound, though less strongly, to GM1 and GD1b. WGA bound to 3'‐nLM1 and 6'‐nLM1, the former twice as strongly as the latter, but not to any sialic acid containing glycolipid of the gangliotetraose series. UEA I showed a high affinity for the Lea glycolipid which has an α 1‐4 linked fucose but not for the glycolipids with αl‐3 or α 1–2 linked fucose. Interestingly, 3'‐nLM1 and nLA1, glycolipids lacking fucose, also bound UEA I. The results show that lectins should be used with caution for establishing terminal sugar sequences in glycosphingolipids.
Role of Mg2+ in the ribozyme system
Authors: Toh Hiroyuki , Imamura Akira , Kanda Katsuya
Published: July 2002

Recently, several kinds of catalytic activities have been identified from L‐19 IVS RNA or ‘ribozyme’. In the catalytic process of ribozyme, Mg2+ is required. We investigated the role of Mg2+ by molecular orbital calculation. Based on the results of the calculation, a model for the self‐splicing reaction was constructed.
A zebrafish homologue of the murine Hox‐2.1 gene
Authors: Njølstad Pål R. , Molven Anders , Fjose Anders
Published: July 2002

Homeobox‐containing sequences were isolated from a genomic library of zebrafish (Brachydanio rerio). A λ clone containing two homeobox cross‐hybridizing regions was characterized. DNA sequencing of one of these regions (ZF‐21) revealed that it contains a homeobox closely related to the Antennapedia class of Drosophila homeobox sequences. Moreover, the deduced amino acid sequence of the C‐terminal end (81 residues including the homeobox) is identical to the corresponding part of the murine Hox‐2.1 protein. Similar to Hox‐2.1, a ZF‐21 derived transcript of 2.3 kb is present in embryos at the somite forming stages.
Apomyoglobin forms a micellar complex with phospholipid at low pH
Authors: Woo Lee Jong , Kim Hyoungman
Published: July 2002

Apomyoglobin was found to break down phosphatidylcholine vesicles to form a micellar complex at pH 4. Size reduction during this process was observed by measuring light scattering at 400 nm and by passing the complex through a gel‐filtration column. Micellar complex formation was confirmed by electron microscopy.
Exon‐intron organization, expression, and chromosomal localization of the human motilin gene
Authors: Yano H. , Seino Y. , Fujita J. , Yamada Y. , Inagaki N. , Takeda J. , Bell G.I. , Eddy R.L. , Fan Y.-S. , Byers M.G. , Shows T.B. , Imura H.
Published: July 2002

The human motilin gene has been isolated and characterized. The gene spans about 9 kilobase pairs (kb) and the 0.7 kb motilin mRNA is encoded by five exons. The 22‐amino‐acid motilin sequence is encoded by exons 2 and 3. The human motilin gene was mapped to the p21.2→p21.3 region of chromosome 6 by hybridization of the cloned cDNA to DNAs from a panel of reduced human‐mouse somatic cell hybrids and by in situ hybridization to human prometaphase chromosomes. RNA blotting using RNA prepared from various regions of the human gastrointestinal tract revealed high levels of motilin mRNA in duodenum and lower levels in the antrum of the stomach; motilin mRNA could not be detected by this procedure in the esophagus, cardia of the stomach, descending colon or gallbladder.
Evidence for interactions of acyl carrier protein with glycerol‐3‐phosphate acyltransferase, an inner membrane protein of Escherichia coli
Authors: Bayan Nicolas , Thérisod Hélène
Published: July 2002

We [(1989) FEBS Lett., in press] have previously shown that membrane vesicles from Escherichia coli contain protein‐binding sites for the acyl carrier protein (ACP). We report now that membrane vesicles prepared from a strain amplified for glycerol‐3‐phosphate acyltransferase (GPAT) contain a higher number of ACP‐binding sites than the membrane vesicles prepared from a wild type strain. In addition, we show that GPAT is retained specifically on an ACP‐Sepharose affinity column and that [3H]ACP binds to the enzyme solubilized by detergent. We conclude that GPAT, an inner membrane protein which catalyses the transesterification of a fatty acyl group from acyl coenzyme A or acyl ACP to glycerol‐3‐phosphate, possesses a binding site for ACP.
Direct sequencing of affinity‐captured amplified human DNA application to the detection of apolipoprotein E polymorphism
Authors: Syvänen Ann-Christine , Aalto-Setälä Katriina , Kontula Kimmo , Söderlund Hans
Published: July 2002

We describe a method for the direct sequencing of DNA amplified by the polymerase chain reaction (PCR). Biotin is introduced into one strand of the amplified DNA using a 5'‐biotinylated PCR primer. The synthezised fragment is captured on an avidin‐matrix and rendered single stranded, whereafter the nucleotide sequence of the immobilized strand is determined by the chain termination method. The method involves few and simple operations and is thus applicable to the analysis of human genes for routine diagnostic purposes. Here we applied the method for determination of the three‐allelic polymorphism of the apolipoprotein E (apo E) gene. We were able to correctly identify the alleles in both homozygous and heterozygous samples.
Synapsin I from human brain
Authors: Severin S.E. , Moskvitina E.L. , Bykova E.V. , Lutzenko S.V. , Shvets V.I.
Published: July 2002

Synapsin I has been isolated from human brain by a rapid and efficient purification technique, and its phosphorylation by human brain Ca2+, phospholipid‐dependent protein kinase (protein kinase C) has been studied. The inhibitory effect of calmodulin on this process has been demonstrated. It is also found that non‐esterified fatty acids and acidic phospholipids are inhibitory for synapsin I phosphorylation by Ca2+, calmodulin‐dependent protein kinase II.
N‐Terminal sequence similarities between components of the multicatalytic proteinase complex
Authors: Lilley Kathryn S. , Davison Matthew D. , Rivett A.Jennifer
Published: July 2002

The multicatalytic proteinase complex is a high molecular weight nonlysosomal proteinase which is composed of many different types of subunit. As part of a study of the possible relationships between subunits, polypeptides derived from the multicatalytic proteinase from rat liver have been subjected to N‐tenninal amino acid sequence analysis. Although several of the subunits are blocked at their N‐termini, sequences have been obtained for 7 of the polypeptides. Each of the 7 sequences is unique but they show considerable sequence similarity, suggesting that the proteins are encoded by members of the same gene family.
Electron microscopy of native and artificial methylreductase high‐molecular‐weight complexes in strain Gö 1 and Methanococcus voltae
Authors: Hoppert M. , Mayer F.
Published: July 2002

The preparation of inside‐out vesicles from methanogenic bacteria with protein cell walls was improved with regard to the preservation of structure and localization of membrane‐bound proteins. Complexes similar to the methanoreductosome in the methanogenic bacterium Gö 1 [1] were also found attached to the inner aspect of the cytoplasmic membrane of Methanococcus voltae. Methanoreductosomes were purified from crude extracts of Gö 1‐cells by affinity chromatography. Under specific conditions at high protein concentrations methyl‐CoM‐methylreductase molecules isolated from Gö 1‐cells could be reassociated to spherical complexes of various sizes, with an appearance similar to that of methanoreductosomes isolated from strain Gö 1.
Analysis of β‐tubulin sequences reveals highly conserved, coordinated amino acid substitutions
Authors: Burns Roy G. , Surridge Christopher
Published: July 2002

Vertebrate β‐tubulins have been classified into six classes on the basis of their C‐terminal sequences [(1987) J. Cell Biol. 105, 1707‐1720]. In particular, the sequences starting at residue 430 differ between isotypes of the same animal but are conserved between species. We extend this analysis and show that there are three ‘hot spots’, at residues 35, 55–57 and 124 which exhibit intra‐species heterogeneity but inter‐species conservation. There is a remarkable correlation between the identity of these residues and the C‐terminal sequences, and suggests that the vertebrate β‐tubulins fall into three broad types. This correlation extends to those non‐vertebrate organisms which have the Type 1 C‐terminal sequence. We propose that these three ‘hot spots’ and the C‐terminal peptide interact in the tertiary structure. We have also noted that the C‐terminal peptide almost always contains a single phenylalanine or tyrosine residue, and that there is a strong correlation between this residue and the amino acids at positions 217/218, in both the vertebrate and non‐vertebrate sequences. We propose that the C‐terminal aromatic amino acid interacts with residues 217/218 in the tertiary structure. Analysis of conditions which stabilise microtubules and/or lower the steady state critical concentration strongly suggests that these two sets of coordinated amino acid substitutions are directly involved in effecting the conformational change associated with GTP hydrolysis which results in dynamic instability. We propose that there is an interaction between the highly acidic sequence between residue 430 and the aromatic amino acid (termed peptide A) and conserved basic amino acids located close to the ‘hot spots’. We suggest that this interaction is altered in response to the assembly‐dependent GTP hydrolysis, with the consequential increase in the subunit dissociation rate constant.
Cyclic nucleotide‐activated channels in the frog olfactory receptor plasma membrane
Authors: Kolesnikov S.S. , Zhainazarov A.B. , Kosolapov A.V.
Published: July 2002

Patch clamp technique was used to record cyclic nucleotide‐dependent current of the frog olfactory receptor cell plasma membrane. Data obtained indicate that the channels passing this current are permeable to Ca2+ or Mg2+ and moderately selective for monovalent cations according to the sequence Li+, Na+, K+ > Rb+ > Cs+ and are effectively blocked by 1‐cis‐diltiazem and 3′,4′‐dichlorobenzamil. The conductance of single cyclic nucleotide‐gated channels in solutions with low Ca2+ and Mg2+ content is about 19 pS. The results demonstrate that cyclic nucleotide‐activated channels of olfactory receptor cells are virtually identical to photoreceptor ones.
Crystallographic investigations of the tryptophan‐derived cofactor in the quinoprotein methylamine dehydrogenase
Authors: Chen Longyin , Mathews F.Scott , Davidson Victor L. , Huizinga Eric G. , Vellieux Frederic M.D. , Duine Johannis A. , Hol Wim G.J.
Published: July 2002

A model of tryptophan tryptophylquinone (TTQ), recently proposed by Mclntire et al. (Science (1991) 252, 817‐824) to be the prosthetic group of the quinoprotein methylamine dehydrogenase, has been compared with electron density maps of this dehydrogenase from Thiobacillus versutus and Paracoccus denitrificans. The comparison shows that the TTQ model can be neatly accommodated, providing strong supportive evidence that TTQ is indeed the cofactor for this group of quinoproteins.
Site‐directed mutagenesis of bovine pancreatic ribonuclease: Lysine‐41 and aspartate‐121
Authors: Trautwein Katrin , Holliger Philipp , Stackhouse Joseph , Benner Steven A.
Published: July 2002

Chemical modification studies suggest that two residues of bovine pancreatic ribonuclease A (RNase A), Lys‐41 and Asp‐121, are important for catalysis. Three mutants of RNase A have been prepared, two point mutants with Lys‐41 altered to Arg‐41 and Asp‐121 altered to Glu‐121, and a double mutant where both residues are altered. The Lys‐41 Arg mutant has ca. 2% the calatylic activity (kcalKm
) of the native protein, while the Asp‐121 Glu mutant has ca. 17% the catalytic activity of the native protein. The double mutant has catalytic activity comparable to the Lys‐41 Arg mutant.
Identification of glycolipid receptors for Helicobacter pylori by TLC‐immunostaining
Authors: Saitoh Toshihito , Natomi Hitomi , Zhao Weilu , Okuzumi Kastuko , Sugano Kentaro , Iwamori Masao , Nagai Yoshitaka
Published: July 2002

Helicobacter pylori has been identified as a causative agent in active chronic gastritis. The receptor for this bacteria, however, is not known. It is likely that the receptor molecules may be glycosphingolipids* as shown in the cases of other bacteria. We explored this possibility by a thin‐layer chromatography (TLC)‐immunostaining method. Among glycosphingolipids extracted from human gastric mucosa, intact Helicobacter pylori specifically bound to PSO3‐GalCer and II3NeuAc‐LacCer, whereas no specific binding to neutral glycosphingolipids, which share the same ceramide moiety with PSO3‐GalCer or II3NeuAc‐LacCer, was demonstrated. Sonicated bacteria could still bind to II3NeuAc‐LacCer with comparable affinity. In contrast, the binding of bacteria of PSO3‐GalCer was greatly diminished upon sonication. These results suggest that each of the oligosaccharide moieties of II3NeuAc‐LarCer and PSO3‐GalCer may be specifically recognized by different ligand molecules of Helicobacter pylori.
Treatment of intact hepatocytes with synthetic diacyl glycerols mimics the ability of glucagon to cause the desensitization of adenylate cyclase
Authors: Newlands Christine , Houslay Miles D.
Published: July 2002

Incubation of intact hepatocytes with either of the synthetic diacyl glycerols 1‐oleoyl‐2‐acetyl glycerol (OAG) or dihexanoyl glycerol (DHG) caused the transient uncoupling of the ability of glucagon to stimulate adenylate cyclase in membranes prepared from those cells. No change occurred in either the activity of the catalytic unit of adenylate cyclase or the coupling of G5 to adenylate cyclase. Diacyl glycerol action appeared to mimic glucagon‐mediated desensitization of adenylate cyclase, suggesting that protein kinase C activation may provide the molecular trigger for glucagon desensitization.
Detection of the lipid‐linked precursor oligosaccharide of N‐linked protein glycosylation in Drosophila melanogaster
Authors: Parker G.F. , Williams P.J. , Butters T.D. , Roberts D.B.
Published: July 2002

The presence of a glycan of the same molecular size as the lipid linked precursor oligosaccharide (Glc3Man9GlcNAc2) of the N‐linked protein glycosylation pathway in mammalian cells has been detected in a glycolipid fraction of cultured Drosophila melanogaster cells. Oligosaccharide sequencing studies were consistent with the existence of a glucosylated high mannose containing structure, which may be the common precursor for N‐linked protein glycosylation in insect cells.
Stimulation of glucose transport in L6 muscle cells by long‐term intermittent stretch—relaxation
Authors: Mitsumoto Yasuhide , Downey Gregory P. , Klip Amira
Published: July 2002

Skeletal muscle stretch increases resting metabolism and causes hypertrophy. We have examined the effect of mechanical stretch in vitro on glucose transport activity and transporter contents in L6 muscle cells. Long‐term (24–48 h) stretch—relaxation (25% maximal elongation at 30 cycles per min) of cell monolayers significantly increased glucose uptake by 1.6‐ to 2‐fold in myotubes but not in myoblasts. The presence of serum was required for the stretch—relaxation induced increase in glucose uptake. Cycloheximide inhibited the mechanical stimulation of glucose uptake, and the latter response was not additive to the stimulatory effect of long‐term exposure to insulin. GLUT1 and GLUT4 glucose transporter contents were not changed in total cell membranes from mechanically stimulated cells relative to controls. These results indicate that mechanical stimulation through passive stretch may be an important regulation of nutrient uptake in fetal myotubes independent of innervation.
Ca2+‐dependent translocation of cytosolic proteins to isolated granule subpopulations and plasma membrane from human neutrophils
Authors: Borregaard Niels , Kjeldsen Lars , Lollike Karsten , Sengeløv Henrik
Published: July 2002

In order to identify cytosolic proteins involved in control of granule exocytosis in human neutrophils, subcellular fractions enriched in each of the 3 major granule subsets were incubated with cytosol from neutrophils in the presence or absence of Ca2+. After washing, proteins were eluted from the organelles by EGTA, Annexins I, II, IV and VI were found to bind to all organelles studied. In addition, a 28‐kDa protein was found to bind exclusively to plasma membranes and secretory vesicles, the most readily exocytosed organelle of neutrophils. Ca2+‐dependent association of cytosolic proteins to different granule subsets may control differential exocytosis of granules.
Engineering firefly luciferase as an indicator of cyclic AMP‐dependent protein kinase in living cells
Authors: Sala-Newby Graciela , Campbell Anthony K.
Published: July 2002

A bioluminescent indicator for protein kinase A has been developed by mutating V217 in firefly (Photinus pyralis) luciferase to R, and the C‐terminal peroxisomal signal removed by PCR. The cDNA for normal and the RRFS mutant luciferase were inserted into pSV7d and expressed in COS‐7 cells. Transient expression in approximately 5% of cells was confirmed by extraction of active luciferase, light emission from cells in the presence of luciferin, and immuno‐localisation. The cyclic‐AMP analogue, 8‐(4‐chlorophenylthio)‐cyclic AMP caused a 5–10% decrease in light emission within 4 min in COS cells expressing the RRFS mutant, but not in cells expressing normal luciferase. This provides for the first time an indicator for detecting and quantifying protein kinase A activation in living cells.
Non‐equivalent natures of the coordinated imidazole rings of cytochrome c3 from D. vulgaris Miyazaki F as studied by 1H NMR
Authors: Akutsu Hideo , Hirasawa Masaaki
Published: July 2002

All of the C2 proton signals of the coordinated histidine residues in the 1H NMR spectrum of cytochrome c
3 from D. vulgaris Miyuzaki F were assigned by specific deuteration. They appeared at extremely high fields and scattered in a wide range from −4 to −22 ppm. This clearly shows that the chemical properties of the imidazole groups are quite different from one another. The extremely high‐field shift of the C2 signal indicates that some of them must carry the imidazolate‐like nature to some extent. This might be responsible for the extremely low redox potentials of the four hemes. On changing temperature, most of them showed Curie‐type change. All of the C2 signals showed a small p2H dependence in the range of p2 4.8–10.0.
Characterization of a new leiurotoxin I‐like scorpion toxin
Authors: Zerrouk H. , Mansuelle P. , Benslimane A. , Rochat H. , Martin-Eauclaire M.F.
Published: July 2002

Three novel peptide inhibitors of the SKCa channels were purified to homogeneity from the venom of the scorpion Androctonus mauretanicus mauretanicus using one step of RP‐HPLC and competition assays with [125I]apamin to rat brain synaptosomes. POi, PO2 and PO5 have K
0.5 of 100,100 and 0.02 nM, respectively, for the apamin binding site. The sequence of PO5 was established and compared to that of other scorpion toxins active on K
+ channels: it contains 31 residues and has a free carboxyl end. It shares sequence similarity with apamin and leiurotoxin I.