bluuebunny/crossref_metadata_2025 huggingface dataset:
Pengembangan Model Pelibatan Orang Tua Dalam Meningkatkan Kreativitas Anak Melalui Pentas Kelas Pada Satuan Paud di Kabupaten Gorontalo
Authors: Sahara La Ode
Published: April 2020

Tujuan penelitian ini adalah untuk: (1) Mendeskripsikan kondisi objektif pelibatan orang tua dalam penyelenggaraan PAUD di Kabupaten Gorontalo. (2) Mendeskrisikan model konseptual pelibatan orang tua dalam meningkatkan kreativitas anak melalui pentas kelas akhir tahun pada satuan PAUD di Kabupaten Gorontalo. (3) Mendeskripsikan implementasi model pelibatan orang tua dalam meningkatkan kreativitas anak melalui pentas kelas akhir tahun pada satuan PAUD di Kabupaten Gorontalo. (4) Mendeskripsikan efektivitas model pelibatan orang tua dalam meningkatkan kreativitas anak melalui pentas kelas akhir tahun pada satuan PAUD di Kabupaten Gorontalo.Pendekatan yang digunakan dalam penelitian ini adalah pendekatan kualitatif dengan jenis penelitian deskriptif. Metode yang digunakan adalah metode research and development (R&D) yang meliputi kegiatan: (1) studi pendahuluan tentang kondisi objektif pelibatan orang tua dalam pendidikan, (2) pengembangan model pendampingan, (3) validasi ahli dan revisi, (4) implementasi model, dan (5) uji keefektifan model.Hasil penelitian pengembangan menunjukkan: (1) Kondisi awal pelibatan orang tua pada pendidikan anak umumnya masih rendah. Hal tersebut dikarenakan pelibatan orang tua belum dilaksanakan secara optimal. (2) Pengembangan model pelibatan orang tua dalam meningkatkan kreativitas anak melalui pentas akhir tahun dilaksanakan melalui perencanaan, pelaksanaan dan pendanaan, melakukan validasi pengembangan model dengan pakar Pendidikan nonformal, praktisi pendidikan nonformal dan teman sejawat, dan menyusun model akhir. (3) Implikasi model pendampingan dilakukan melalui kegiatan uji coba dan uji aplikasi model pelibatan. Keseluruhan kegiatan tersebut dilakukan melalui tahapan; input, proses, output dan outcome. (4) Pengembangan model pelibatan orang tua memberikan dampak pada peningkatan kreativitas dan pelaksanaan pentas akhir tahun. Temuan penelitian ini menunjukkan model pelibatan orang tua dalam meningkatkan kreativitas anak melalui pentas akhir tahun yang dikembangkan direkomendasikan kepada pihak-pihak terkait dalam meningkatkan pengelolaan PAUD di Provinsi Gorontalo.
Meningkatkan Motivasi Guru Menciptakan Media Pembelajaran Melalui Teknik Pendampingan di TK Tunas Harapan Kota Timur
Authors: Alhasni Salma
Published: April 2020

Masalah dalam penelitian tindakan kelas ini adalah Apakah dengan penerapan Teknik Pendampingan dapat meningkatkan motivasi guru menciptakan media pembelajaran di TK Tunas Harapan Kota Gorontalo?. Adapun tujuan penelitian ini adalah untuk meningkatan motivasi guru menciptakan media pembelajaran melalui penerapan Teknik Pendampingan di TK Tunas Harapan. Penelitian ini merupakan penelitian tindakan sekolah dengan metode analisis deskriptif yang meliputi siklus I dan siklus II, dengan langkah perencanaan, tindakan, observasi, dan refleksi. Hasil siklus I menunjukan dari 2 orang guru yang dilatih melalui proses pendampingan, diperoleh bahwa 1 orang atau 50% memiliki motivasi menciptakan media pembelajaran kemudian mengalami peningkatan pada siklus II yaitu dari 2 orang guru, diperoleh bahwa 100% telah memiliki motivasi menciptakan media pembelajaran bagi siswa TK. Dengan demikian indikator kinerja dikatakan berhasil. Simpulan dari penelitian ini adalah Teknik Pendampingan dapat meningkatkan motivasi guru menciptakan media pembelajaran di TK Tunas Harapan
Pengaruh Kemandirian Belajar Siswa Terhadap Prestasi Belajar di SDN 03 Limboto Barat Kabupaten Gorontalo
Authors: Uki Finartin , Ilham Asni
Published: April 2020

Permasalahan yang menjadi fokus penelitian ini adalah rendahnya prestasi belajar siswa SDN 03 limboto Barat. Penelitian ini dilaksanakan di SDN 03 Limboto Barat dengan tujuan  untuk mengetahui pengaruh kemandirian belajar siswa terhadap prestasi belajar. Penelitian ini menggunakan pendekatan kuantitatif. Teknik pengumpulan data yang digunakan adalah angket dan studi dokumen. Teknik analisis data yang digunakan ialah analisis regresi dan korelasi. Anggota populasi pada penelitian ini berjumlah 130 orang, sampel yang digunakan diambil dari populasi tersebut dengan menggunakan teknik Proportionate Sratified Random Sampling yaitu 34 siswa. Berdasarkan hasil penelitian diperoleh  bentuk persamaan regresi linier sederhana Ŷ = 34,96 + 0,58X sehingga dapat disimpulkan variabel X (Kemandirian Belajar) diikuti variabel Y (Prestasi belajar siswa). Kemudian dari hasil perhitungan koefisien korelasi diperoleh r= 0,852 dengan koefisien determinasi r2= 0,7259. Uji signifikan koefisien korelasi memperoleh hasil perhitungan diperoleh harga t hitung sebesar = 9,22 sedangkan dari daftar distribusi t pada taraf 5% diproleh t daftar = 1,69. Ternyata harga t hitung lebih besar dari t daftar, atau harga t hitung berada di luar daerah penerimaan Ho. Sehingga dapat disimpulkan bahwa Ho ditolak dan menerima Ha, dengan demikian dapat disimpulkan bahwa terdapat pengaruh antara kemandirian belajar dengan prestasi belajar siswa.
Role of Social Media in COVID-19 Pandemic
Authors: Hussain Wajahat
Published: April 2020

Coronavirus disease 2019 (COVID-19) pandemic has produced a global health crisis that has had a deep impact on the way we perceive our world and everyday lives. Not only the spread rate of contagion and patterns of transmission endangered our sense of security, but the safety measures put in place to contain the spread of the virus also require social distancing by refraining from doing what is inherently human, which is to find comfort in the company of others. Within this context of physical threat, social and physical distancing, the role of the different mass media channels and social media in lives on individual, social and societal levels cannot be underestimated.
Universality of Social Health
Authors: Chakravorty Indranil
Published: April 2020

As you read the articles that I have had the pleasure to curate for this special edition of SUSHRUTA, published along with the #BAPIOAC19 conference in London this wet and frosty November, you will see glimmers of hope and aspirations of a broad-church of people that make up the health service. NHS professionals come from all over the world, bringing their cultures from distant shores that were once touched by the British Empire (now the Commonwealth) You will also witness the ambitious plans for future proofing what is arguably the best universal health system in the world. It is a time for 20/20 vision; we recognise not only the challenges that face the UK population and the NHS but we also envisage how the strengths of the diverse multi-skilled workforce come together in making the UK NHS star shine even brighter.
Pharmacist as an Integral Member of the Medical Team:
Authors: Hackett Aine
Published: April 2020

Pharmaceutical products account for the third largest spend for the National Health Service (NHS), with the bill in England alone, exceeding £17 billion per year 1 . The majority of patients admitted into hospital take medicines or will have done so at some point prior to their admission. So, predictably pharmacy and in particular the pharmacist in the NHS 10-year plan, is defined as having “an essential role to play” 2 .
A Critical Review on Haratala (An Arsenical Compound)
Authors: Wadnerwar Nilima N , Rajput Dhiraj Singh , Deshmukh Akhilesh A , Gaikwad Apurva
Published: April 2020

Background:

Haratala is described as the Dhatu visha and it contains arsenic and sulfur. Arsenic is a heavy metal that may lead to acute or chronic heavy metal toxicity. Although Haratala is effective and popular as Rasamanikya, it is not used as Rasayana or therapeutic drug in routine practice either in pure or in Bhasma form. It is used as a main drug or an auxiliary drug to prepare formulation.

Aim:

To focus on the various utilities of Haratala, so that it can be safely used in the clinical practice in a wide range of indications.

Review results:

In the literature available, it is observed that along with therapeutic utility of Haratala, Haratala Bhasma is used as Rasayana. But arsenic present in Haratala may cause toxicity if shodhana and marana of Haratala are not conducted properly. It is observed from the previous research that shodhana may enhance the synergistic effect of Haratala in cellular apoptosis for the treatment of leukemia. Marana may provide safer bioassimilability before their use in most of the formulations in which it may act as an antagonist and subside the toxicity of the formulations.

Conclusion:

Haratala Bhasma is indicated in various disease conditions. Hence, it can be used as Avasthika Rasayana. But during its use, safety of the drug should be ensured by following proper shodhana and marana procedures.

Clinical significance:

Studies should be conducted to observe its efficacy in healthy individuals as Rasayana and in patients of acute promyelocytic leukemia (APL) as an adjuvant drug.
A Comprehensive Review of Karavīra
Authors: Patel Vivek K , Acharya Rabinarayan , Patel Bhupesh R
Published: April 2020

Background:

Karavīra, though considered as Upviṣa, has been a part of Ayurvedic pharmacopoeia for treating many diseases. As a drug, karavīra and its various parts have been utilized in different dosage forms against various disease conditions through external and internal application. Classical texts are written at different time periods, since the origin of Āyurveda. These information have not been critically reviewed and published in a compiled format. A systematic review regarding the identification and use of this classical drug is the need of time.

Aim:

To have a comprehensive review on karavīra like classification, synonyms, pharmacological properties, actions, varieties, and formulations containing karavīra or its parts as an ingredient from different classical texts and to present them in a systematic manner.

Results:

After review, different opinions are found about the types of karavīra based on their flower color. Various synonyms are attributed to describe its morphological as well as pharmacological characters. Ayurvedic classics classified karavīra in varga based on various criteria. karavīra has been used as an ingredients in 222 formulations having external and internal uses. Nerium oleander L. and Thevetia peruviana (Pers.) K. Schum. are source plants of karavīra reported for their therapeutic value and various pharmacological activities. All parts of these plants are having toxic properties because of their phytoconstituents present in them.

Conclusion:

karavīra described in classical texts of Āyurveda by using various synonyms, their varieties, varga, their rasapancaka, karma, and a wide range of therapeutic importance in 222 formulations to combat 62 diseased conditions. Maximum formulations are indicated in the treatment of Kuṣṭha. Maximum dosage forms are Taila. External application of karavīra are found more as compared to internal application, which may be due to its poisonous character.
Observation on Single-plant Therapy (Ekala Dravya Chikitsa) among Folk Healers of Andaman and Nicobar Islands, India
Authors: Mane Santosh S , Sinimol Thekkekkootthumughath P , Masram Pravin R , Rath Chinmay , Susmitha Bonthu , Gaur Sugriv K , Srikanth Narayanam , Tripathi Ashish Kumar , Kamble Mayur Y , Rattna Joseph GV
Published: April 2020

Introduction:

The collection and documentation of folklore claims of the native population of Andaman and Nicobar Islands including tribes are very essential as these claims can pave the path for new drug discovery. The concept of treating a disease by using a single plant or utilizing it as health supplements are rapidly spreading all over the world and it is widely practiced by physicians.

Objectives:

This study sought to document the distinct plant species used in treatment by the native and tribes of Andaman and Nicobar Islands, India.

Materials and methods:

During the year 2013 to 2016, a total of 23 periodical surveys of 75 forest beats of Andaman and Nicobar Islands were conducted and 62 local traditional folk healers were interviewed as per the questionnaire based on ethnomedicinal, survey protocol.

Observations:

One hundred and forty-one folklore claims related to single-herbal remedies were registered which comprises 103 medicinal plant species used to treat 47 different diseases. Maximum numbers of species, i.e., 13 are utilized to cure abdominal pain followed by 12 species for headache and 11 species for fever.

Discussion:

Among the 103 plant species, 85 species have been quoted in the texts of Ayurveda and the remaining 18 plants have been also defined as folk medicine in other literature of plant studies. However, this study highlights their utilization in common disease by the native and tribal folk healers of Andaman and Nicobar Islands.

Conclusion:

This study gives immense output on the use of the individual medicinal plant in varied systemic and local diseases from the folk healers of the Andaman and Nicobar Islands. Organized and constructive information incorporated in this article may aid in conducting further pharmacological, toxicological, and clinical studies on reported claims to confirm their therapeutic efficacy and utility.
Corporate Greenwashing and Canada Goose: Exploring the Legitimacy–Aesthetic Nexus
Authors: Gacek James
Published: April 2020

Public discourse on environmental responsibility and sustainability continues to pressure corporations, especially those that have been portrayed as key contributors of environmental harm. Greenwashing is a strategy that companies adopt to engage in symbolic communications with environmental issues without substantially addressing them in actions. This paper aims to raise awareness of corporate greenwashing, drawing attention to issues that progress the trend of individualized responsibility and consumption, while concealing the social and (eco)systemic issues in the process. By drawing on the case study of winter apparel company Canada Goose, this paper questions whether businesses can ‘go green’ in good faith, if corporate responsibility and environmental responsibility can ever be reconciled, and if there is considerable need to clarify the intended effects and unintended consequences of corporate greenwashing.
Improving the Penile Skin Adequacy and Appearance by Patching of the Preputial Skin Island Flap (PSIF) in Distal and Midshaft Hypospadias Associated with Severe Ventral Skin Deficiency: Experience in 224 Cases
Authors: Molaeian Mansour , Eskandari Farid , Raji Hojattollah , Ghavami-Adel Maryam , Mollaeian Arash
Published: April 2020

Objectives: Reconstruction of ventral penile skin, in midshaft and distal hypospadias is problematic in many cases of severe ventral dysplasia. So we plan to use the tailored preputial skin island flap (PSIF) to cover the ventral defect. Methods: In this prospective cohort study 224 boys with midshaft and distal hypospadias associated with sever ventral dysplasia were operated from March 2009 until January 2016. The large U-shaped bare area which exposed on the ventrum after release of curvature was quilted by the PSIF as a patch. In each case, the variable size of the flap was required. The prepuces without the predominant artery were excluded from the study due to poor vasculature. The patients followed for 8 months to 7years. Results: Flap ischemia developed in 7 initial cases (3.125%), all of which subsequently faded. Some degree of ischemic discoloration was encountered at margins of the flap (12 cases, 5.3%), which resolved spontaneously. In 5 out of 224 patients (2.23 %), the flap persisted as a hypertrophied area of skin in the long-term follow-up. All of them needed to be corrected surgically. Near normal appearing penile skin was accomplished in the majority of patients during the follow-up. However, lack of the median raphe in the ventrum was evident in all. Conclusions: Tailored preputial skin island flap, as a native tissue to cover the exposed large bare area on the ventrum in cases of the distal and mid hypospadias with severe ventral dysplasia may improve the appearance and adequacy of the penile skin. It provides room for full erection and improves the patient’ s perception of body image and self-esteem.
Microscale Mapping of Structure and Stress in Barium
Titanate
Authors: Howell Jane A. , Vaudin Mark D. , Friedman Lawrence H. , Cook Robert F.
Published: April 2020

Cross-correlation of electron backscatter diffraction (EBSD) patterns was used to
generate rotation, strain, and stress maps of single-crystal tetragonal barium titanate
(BaTiO3) containing isolated, small, sub-micrometer a domains separated from a c-domain
matrix by 90° domain boundaries. Spatial resolution of about 30 nm was demonstrated over
5 μm maps, with rotation and strain resolutions of approximately 10−4. The magnitudes of
surface strains and, especially, rotations peaked within and adjacent to isolated
domains at values of approximately 10−2, i.e., the tetragonal distortion of BaTiO3. The
conjugate stresses between a domains peaked at about 1 GPa, and principal stress
analysis suggested that stable microcrack formation in the c domain was possible. The
results clearly demonstrate the applicability of EBSD to advanced multilayer ceramic
capacitor reliability and strongly support the coupling between the electrical
performance and underlying mechanical state of BaTiO3-containing devices.
Antibodies to the calmodulin‐binding Ca2+‐transport ATPase from smooth muscle
Authors: Wuytack Frank , Schutter Greet De , Verbist Jan , Casteels Rik
Published: July 2002

Antibodies were raised against a calmodulin‐binding CaMg‐ATPase (Ca2+‐transport ATPase) from smooth muscle. The binding of these antibodies to a number of related Ca2+‐transport ATPases was studied. Antibodies to the calmodulin‐binding ATPase from porcine antrum (stomach) smooth muscle do not only bind to this CaMg‐ATPase, but also to the corresponding enzyme in porcine erythrocytes. However, they do not bind to the CaMg‐ATPase from sarcoplasmic reticulum of porcine skeletal muscle. The binding of these antibodies to the CaMg‐ATPase of smooth muscle, does not inhibit the enzyme activity.
Multi‐site phosphorylation in ox‐kidney branched‐chain 2‐oxoacid dehydrogenase complex
Authors: Lau Kim S. , Phillips Caroline E. , Randle Philip J.
Published: July 2002

Tryptic [32P]phosphopeptides were prepared from [32P]phosphorylated ox‐kidney branched‐chain complex and analysed by high‐voltage paper electrophoresis at pH 1.9. In the maximally phosphorylated complex 3 tryptic [32P]phosphopeptides were identified (TA, TB, TC). R
F‐values relative to N
6‐dinitrophenyllysine were (mean ± SEM for 25 obs.): TA, 1.53 ± 0.03; TB, 1.07 ± 0.02; TC, 0.65 ± 0.01. Relative rates of phosphorylation were TA > TB > TC. Phosphorylation of TA reached a maximum when about 66% of the complex was inactivated. Phosphorylation of TB and TC was associated mainly with 66–95% inactivation of the complex.
Lack of ability of trypsin‐treated mitochondrial F1‐ATPase to bind the oligomycin‐sensitivity conferring protein (OSCP)
Authors: Hundal Torill , Norling Birgitta , Ernster Lars
Published: July 2002

Soluble beef‐heart mitochondrial F1‐ATPase modified in its α‐subunit by mild trypsin treatment (α′‐F1) can no longer bind oligomycin‐sensitivity conferring protein (OSCP) but is still capable of binding to F1‐depleted submitochondrial particles, giving rise to a maximally oligomycin‐sensitive ATPase, provided the particles contain their native complement of OSCP. When OSCP is removed from the particles, α′‐F1 can still bind to the particles, but added OSCP induces only a low degree of oligomycin sensitivity. The possible role of OSCP in the functional coupling of the catalytic (F1) and H+‐translocating (Fo) moieties of mitochondrial ATPase is discussed. The results suggest a functional similarity between the OSCP component of mitochondrial ATPase and the δ‐subunit of E. coli ATPase, which is in accordance with the structural homology recently found to exist between the two polypeptides.
Specific leukotriene formation by purified human eosinophils and neutrophils
Authors: Verhagen Jan , Bruynzeel Pieter L.B. , Koedam Johannes A. , Aryan Wassink G. , de Boer Martin , Terpstra Gerben K. , Kreukniet Johannes , Veldink Gerrit A. , Vliegenthart Johannes F.G.
Published: July 2002

Human granulocytes isolated from peripheral blood have been described to synthesize both LTB4 and LTC4 from arachidonic acid. We have observed that the amount of LTC4 produced by human granulocyte preparations is strongly dependent on the relative amount of eosinophils. To investigate a possibly significant difference in leukotriene synthesis of the eosinophilic and neutrophilic granulocytes, we developed a purification method to isolate both cell types from granulocytes obtained from the blood of healthy donors. Leukotrienes were generated by incubation of the purified cells with arachidonic acid, calcium ionophore A23187, calcium‐chloride and reduced glutathione. Surprisingly, eosinophils were found to produce almost exclusively the spasmogenic LTC4. In contrast, neutrophils produce almost exclusively the chemotactic LTB4, its ω‐hydroxylated metabolite 20‐hydroxy‐LTB4 and two non‐enzymically formed LTB4 isomers.
Cotransport of proline and Li+ in Escherichia coli
Authors: Tsuchiya Tomofusa , Yamane Yoshiko , Shiota Sumiko , Kawasaki Takashi
Published: July 2002

Uptake of Li+ induced by the addition of proline to a cell suspension of Escherichia coli was detected using an Li+‐selective electrode. This Li+ uptake was inhibited by L‐azetidine 2‐carboxylic acid, a competitive inhibitor of the proline transport system. Thus, direct evidence for Li+‐proline cotransport via the proline transport system was obtained. Kinetic parameters of the Li+ uptake were determined.
12‐O‐Tetradecanoylphorbol 13‐acetate stimulates inositol lipid phosphorylation in intact human platelets
Authors: de Chaffoy de Courcelles D. , Roevens P. , van Belle H.
Published: July 2002

Potyphosphoinositide
Phorbol ester
Protein kinase C
Binding properties of a novel calcium channel activating dihydropyridine in monolayer cultures of beating myocytes
Authors: Bellemann Peter
Published: July 2002

Binding characteristics of [3H]BAY K 8644, a new class of pharmacologically potent compounds, the calcium channel activating dihydropyridines (DHP), were demonstrated in cultured myocardial cells. [3H]BAY K 8644 exhibited reversible and saturable binding to myocytes, and specific binding was Ca2+‐dependent. The equilibrium dissociation constant, K
d, was 35.2 nM, and maximal binding capacity, B
max, was 1.07 pmol/mg protein. Binding of the 3H‐ligand was highly specific for various potently displacing DHP derivatives (either the calcium channel activating BAY K 8644, or the Ca2+ entry blockers of the nifedipine type) with inhibition constants (K
i values) in the nanomolar range. BAY K 8644, on the other hand, showed very low affinity to other receptors tested in brain and heart membranes. Displacement potency of BAY K 8644 correlated well with data of the functional pharmacology; e.g., the enhanced myocardial contractility. Results from competition studies using [3H]BAY K 8644 and [3H]nimodipine support the conclusion that both the channel activating and inhibiting DHP structures interact with the same specific receptor site that might be associated with the putative Ca2+‐channel.
Biosynthesis of oxytocin in the corpus luteum
Authors: Swann R.W. , O'Shaughnessy P.J. , Birkett S.D. , Wathes D.C. , Porter D.G. , Pickering B.T.
Published: July 2002

Oxytocin biosynthesis
Corpus luteum
Bovine luteal cell
Ovine luteal cell
Oxytocin precursor
Kainate‐induced uptake of calcium by synaptosomes from rat brain
Authors: Pastuszko Anna , Wilson David F.
Published: July 2002

Kainic acid induces a rapid increase in 45Ca2+ uptake by crude synaptosomal fractions isolated from rat brain. This enhanced Ca2+ permeability occurs with a half‐time of approx. 1 s, similar to the fast phase of depolarization‐induced calcium uptake. The depolarization‐induced uptake of calcium is inhibited 85% by 3 mM CoCl2 80% by 100μM quinacrine and 50% by 15μM trifluoperazine while these agents had little effect on the kainate‐induced uptake. It is proposed that kainate induces receptor‐mediated opening of a class of calcium channels with properties different from those of the voltage‐dependent channels.
Pertussis toxin inhibits chemotactic factor‐induced phospholipase C stimulation and lysosomal enzyme secretion in rabbit neutrophils
Authors: Bradford Peter G. , Rubin Ronald P.
Published: July 2002

Pertussis toxin suppressed [32P]polyphosphoinositide breakdown and lysosomal enzyme secretion induced by fMet‐Leu‐Phe in rabbit neutrophils. Likewise, fMet‐Leu‐Phe‐ or leukotriene B4‐evoked (3H]inositol trisphosphate accumulation was inhibited by the toxin. These findings, taken together with evidence that pertussis toxin specifically causes inactivation of the guanine nucleotide binding protein (Ni), suggests that guanine nucleotide binding proteins may mediate coupling between calcium‐mobilising receptors and phospholipase C‐mediated reactions in rabbit neutrophils.
Stimulation of guanylate cyclase by atrial natriuretic factor in isolated human glomeruli
Authors: Ardaillou Nicole , Nivez Marie-Paule , Ardaillou Raymond
Published: July 2002

Atrial natriuretic factor
Isolated glomerulus
Guanylate cyclase
Cyclic GMP
Pertussis toxin selectively abolishes hormone induced lowering of cytosolic calcium in GH3 cells
Authors: Schlegel Werner , Wuarin François , Zbaren Claude , Wollheim Claes B. , Zahnd Gaston R.
Published: July 2002

Pertussis toxin
Cytosolic Ca2+

Somatostatin
Muscarinic cholinergic agonist
TRH
Pituitary
Three‐dimensional structure of renal Na,K‐ATPase determined by electron microscopy of membrane crystals
Authors: Herbert Hans , Skriver Elisabeth , Maunsbach Arvid B.
Published: July 2002

The three‐dimensional structure of Na,K‐ATPase was determined by electron microscopy and image processing. Tilt series of negatively stained membrane crystals were recorded. The projections were analyzed by Fourier methods and the data combined to a 3‐D model. The unit cell contains two rod‐shaped stain‐deficient regions interpreted as αβ‐protomers of Na,K‐ATPase. The rods are related by dyad axes oriented perpendicular to the membrane. Outside the lipid bilayer the rods contact different protein units on the two sides of the membrane.
Differentiation of [3H]phencyclidine and ( + )‐[3H]SKF‐10,047 binding sites in rat cerebral cortex
Authors: Goldman Mark E. , Jacobson Arthur E. , Rice Kenner C. , Paul Steven M.
Published: July 2002

SKF‐10,047
Sigma receptor
Phencyclidine
Phencyclidine receptor
Psychotomimetic activity
N‐α‐Tosyl‐L‐lysine chloromethyl ketone and N‐α‐tosyl‐L‐phenylalanine chloromethyl ketone inhibit protein kinase C
Authors: Solomon David H. , O'Brian Catherine A. , Bernard Weinstein I.
Published: July 2002

Protein kinase C
Tumor promotion
Protease inhibitor
The activation of protein kinase C by the polyphosphoinositides phosphatidylinositol 4,5‐diphosphate and phosphatidylinositol 4‐monophosphate
Authors: O'Brian Catherine A. , Arthur W.Lane , Weinstein I.Bernard
Published: July 2002

Protein kinase C(PKC) is a Ca2+‐ and phospholipid‐dependent protein kinase which can be activated by diacylglycerol, a product of polyphosphoinositide hydrolysis. In this report, we show that the polyphosphoinositides L‐α‐phosphatidylinositol 4‐monophosphate (PI 4P) and L‐α‐phosphatidylinositol 4,5‐diphosphate (PI 4,5DP) can serve as phospholipid cofactors of isolated rat brain PKC. The order of potency of the phosphoinositides in the activation of PKC, PI > PI 4P > PI 4,5DP, shows a negative correlation with the degree of acidity of the phospholipid head group, whether 1 mM Ca2+ or 200 nM TPA is present in the reaction assay mixture. Although the polyphosphoinositides are by themselves weaker activators of PKC than PI, small amounts of PI 4,5DP cause a two‐fold enhancement of PKC in the presence of Ca2+ and PI. While the endogenous phospholipid cofactors of PKC remain to be identified, these results suggest that the small amounts of polyphosphoinositides which are present in cell membranes may play a direct role in the activation of PKC in vivo, by serving as phospholipid cofactors of the enzyme.
Localization of mRNAs encoding the α‐subunits of signal‐transducing G‐proteins within rat brain and among peripheral tissues
Authors: Brann Mark R. , Collins Regina M. , Spiegel Allen
Published: July 2002

The sequence of the mRNAs which encode the α‐subunits of the signal‐transducing G‐proteins Gs, Go and two forms of Gi (termed Gi1 and Gi2) have recently been reported. Based on rat sequences we prepared oligodeoxynucleotide probes for measurement of these mRNAs in rat brain and peripheral tissues. The relative abundance of these mRNA species in brain was Gs>Go ∼ Gi2>Gi1. The Gs and Gi2 mRNAs had somewhat lower levels in heart, kidney and liver than in brain, and Go and Gi1 mRNAs were not detected in the peripheral tissues. Using in situ hybridization we localized each of these mRNAs within slices of the rat brain. The patterns of distribution of Gs and Gi2 mRNA were very similar, but very different from that of Go and Gi1 mRNA. These data illustrate that receptor‐effector coupling G‐proteins are regionally specialized in their expression. This regional specialization may reflect a selective coupling of individual G‐proteins with the various neurotransmitter receptors and effector pathways.
Identification of the lesion in the stimulatory GTP‐binding protein of the uncoupled S49 lymphoma
Authors: Rall Thomas , Harris Bruce A.
Published: July 2002

The stimulatory GTP‐binding protein (Gs) of the uncoupled mutant of S49 lymphoma cells is deficient in its ability to transduce hormonal signals from ligand‐bound β‐adrenergic receptors to the catalytic component of adenylate cyclase. In order to define the genetic defect in the Gs of uncoupled S49 cells, a complementary DNA clone encoding the α‐subunit of Gs was analyzed and the deduced primary structure of the defective subunit compared to that of the wild‐type subunit. A single nucleotide transversion was found that coded for a proline rather than an arginine at residue 389. The results indicate a domain of the α‐subunit of Gs that specifically interacts with hormone receptors.
Isolation of succinate dehydrogenase from Desulfobulbus elongatus, a propionate oxidizing, sulfate reducing bacterium
Authors: Samain E. , Patil D.S. , DerVartanian D.V. , Albagnac G. , LeGall J.
Published: July 2002

Succinate dehydrogenase was purified from the particulate fraction of Desulfobulbus. The enzyme catalysed both fumarate reduction and succinate oxidation but the rate of fumarate reduction was 8‐times less than that of succinate oxidation. Quantitative analysis showed the presence of 1 mol of covalently bound flavin and 1 mol of cytochrome b per mol of succinate dehydrogenase. The enzyme contained three subunits with molecular mass 68.5, 27.5 and 22 kDa. EPR spectroscopy indicated the presence of at least two iron sulfur clusters. 2‐Heptyl‐4‐hydroxy‐quinoline‐N‐oxide inhibited the electron‐transfer between succinate dehydrogenase and a high redox potential cytochrome c
3 from Desulfobulbus elongatus.
On the role of N‐7‐mercaptoheptanoyl‐O‐phospho‐L‐threonine (component B) in the enzymatic reduction of methyl‐coenzyme M to methane
Authors: Ellermann J. , Kobelt A. , Pfaltz A. , Thauer R.K.
Published: July 2002

The reduction of methyl‐coenzyme M (CH3SCoM) to methane in methanogenic bacteria is dependent on component B (N‐7‐mercaptoheptanoyl‐O‐phospho‐L‐threonine, HSHTP). We report here that S‐methyl‐component B (N‐7‐(methylthio)heptanoyl‐O‐phospho‐L‐threonine, CH3SHTP) can substitute for neither CH3SCoM nor HSHTP in the methyl‐CoM reductase reaction. Rather, CH3SHTP proved to be an inhibitor competitive with HSHTP (apparent K
i=6 μM) and noncompetitive with CH3SCoM. These results make it very unlikely that HSHTP functions as a methyl group carrier. A role for HSHTP as direct electron donor for CH3SCoM reduction to CH4is proposed.
Inositol 1,3,4,5‐tetrakisphosphate increases the duration of the inositol 1,4,5‐trisphosphate‐mediated Ca2+ transient
Authors: Joseph Suresh K. , Hansen Carl A. , Williamson John R.
Published: July 2002

The effect of Ins 1,3,4,5‐P4 on the intracellular Ca2+ mobilization produced by Ins 1,4,5‐P, has been examined in permeabilized hepatocytes. Ins 1,3,4,5‐P4 did not affect the magnitude of the Ins 1,4,5‐P3‐mediated Ca2+ release but did inhibit re‐accumulation of the released Ca2+ back into intracellular stores. This effect was not mimicked by Ins 1,3,4‐P3. In hepatocytes, the re‐uptake phase of the response results from Ins 1,4,5‐P3 hydrolysis. Measurements using labeled substrates indicate that Ins 1,3,4,5‐P4 inhibits the hydrolysis of Ins 1,4,5‐P3 and vice versa. Since the removal of the 5‐phosphate on Ins 1,4,5‐P3 and Ins 1,3,4,5‐P4 is a common step in the disposal of both compounds, it is suggested that one of the biological effects of Ins 1,3,4,5‐P4 may be to slow hydrolysis of Ins 1,4,5‐P3 and thereby prolong the duration of a Ca2+ transient.
The presence of the 30 nm filament structure of chromatins in intact chicken erythrocytes observed by 31P NMR
Authors: Nishimoto Shozoh , Akutsu Hideo , Kyogoku Yoshimasa
Published: July 2002

A 31P NMR spectrum of chicken erythrocyte chromatins in vivo was obtained. The spectrum was characterized by examining the spectra of isolated nuclei and nondigested chromatins under various conditions. The results have shown that most chromatins in chicken erythrocyte nuclei assume the 30 nm filament structure as a second step of condensation of DNA in vivo.
Purification of a novel heat‐stable translational inhibitor from rabbit reticulocyte lysates
Authors: Gaitero F. , Limas G.G. , Mendez E. , de Haro C.
Published: July 2002

We have purified to apparent homogeneity a novel heat‐stable (HS) factor from postribosomal supernatants of rabbit reticulocyte lysates by heating for 10 min at 80°C, fractionation on Sephadex, anion‐exchange chromatography on QMA Accell, and gel filtration HPLC. The apparent molecular mass of HS is 500–1000 Da on the basis of its behaviour on gel filtration. Like a factor from bovine heart [(1982) Proc. Natl. Acad. Sci. USA 79, 3134–3137], the reticulocyte HS inhibits translation in hemin‐supplemented lysates with biphasic kinetics similar to hemin deficiency and promotes phosphorylation of the α‐subunit of the eukaryotic initiation factor eIF‐2. It is active at nanomolar concentrations. Reticulocyte HS appears to be neither a peptide nor an oligonucleotide since HS activity was insensitive to proteolytic or nucleolytic digestion.
Intracellular calcium release mediated by two muscarinic receptor subtypes
Authors: Neher Erwin , Marty Alain , Fukuda Kazuhiko , Kubo Tai , Numa Shosaku
Published: July 2002

Four subtypes of muscarinic acetylcholine receptor (mAChR) were stably expressed in neuroblastoma‐glioma hybrid cells (NG108‐15). By combining fluorescent indicator dye (fura‐2) studies with electrophysiological measurements it is shown that stimulation of mAChR I and mAChR III readily leads to release of calcium from intracellular stores and to associated conductance changes, whereas stimulation of mAChR II and mAChR IV exerts no such effect. Dose‐response curves describing the amplitude or the delay of the calcium rise induced by acetylcholine suggest that the apparent affinity of mAChR III for its agonist is higher by about one order of magnitude than that of mAChR I. Ionic substitution experiments and current fluctuation analysis indicate that calcium activates a K+‐specific conductance of ‘small’ single‐channel amplitude similar to the SK type [1]. Furthermore, an outward current (M current) suppressed by activation of mAChR I and mAChR III has a single‐channel amplitude corresponding to a conductance of approximately 3 pS.
Isolation and identification of hydroxyproline analogues of bradykinin in human urine
Authors: Kato Hisao , Matsumura Yasuhiro , Maeda Hirosi
Published: July 2002

Hydroxyproline (Hyp) analogues of bradykinin and lysyl‐bradykinin, in which the third residue of bradykinin, proline, is replaced by hydroxyproline, were isolated from human urine. Their amino acid sequences were confirmed by both amino acid and sequence analyses, and also by comparison of their chromatographic behavior with that of synthetic peptides. The possibility that Lys‐Ala3‐bradykinin, isolated by Mindroiu et al. [(1986) J. Biol. Chem. 261, 7407‐7411] from human urine, was actually Lys‐Hyp3‐bradykinin is discussed.
Monoclonal antibodies to type IV collagenase recognize a protein with limited sequence homology to interstitial collagenase and stromelysin
Authors: Höyhtyä Matti , Turpeenniemi-Hujanen Taina , Stetler-Stevenson William , Krutzsch Henry , Tryggvason Karl , Liotta Lance A.
Published: July 2002

Type IV collagenase is a metalloproteinase associated with metastatic tumor cells. It specifically cleaves the triple helical basement membrane (type IV) collagen molecule at a single site. Monoclonal antibodies which block the activity of the human type IV collagenase were developed and used to purify this antigen. The purified type IV collagenase was partially sequenced following cyanogen bromide and trypsin cleavage. The amino acid sequence of the human type IV collagenase fragments revealed a region homologous to the human interstitial collagenase and stromelysin. However, several sequences in type IV collagenase were identified which are distinct from the latter. Polyclonal antibodies were raised against a synthetic peptide derived from such a sequence. Following affinity purification, the antibodies recognized the denatured human type IV collagenase in Western immunoblotting. These data indicate that type IV collagenase is a distinct member of a general family of metalloproteinases.
Early reduction of cytochrome c in hypoxia
Authors: Chance B.
Published: July 2002

Shifts of the steady state of cytochromes a, a
3 and c at high pO2 values are cited as evidence of the low O2 affinity of cytochrome oxidase in vivo [(1971) Brain Res. 108, 143–154; (1985) Adv. Exp. Med. Biol. 191, 833–842; (1987) in: Int. Soc. Oxygen Transp. Tissue, Sapporo, p. 84]. Highly aerobic, small diploid yeast cells show <0.44% change of steady state in the interval prior to an abrupt reduction of cytochrome c. Thus, ‘pre‐reduction’ seems falsified as a physiological response of metabolizing yeast cells that are free of hemoglobin, do not aggregate and maintain a steady state. Pre‐reduction may be due to spectroscopic interference, for example hemoglobin deoxygenation, to oxygen gradients in aggregated cells and tissues or to non‐steady states of substrate and metabolic controls, as contrasted to an altered cytochrome oxidase oxygen affinity in vivo.
Stabilisation of cathepsin E by ATP
Authors: Thomas D.J. , Richards A.D. , Jupp R.A. , Ueno E. , Yamamoto K. , Samloff I.M. , Dunn B.M. , Kay J.
Published: July 2002

The hydrolysis of 3 distinct substrates by cathepsin E from human red blood cells and gastric mucosa was measured in the presence and absence of physiologically relevant concentrations of ATP. At pH values below about 5.0, the nucleotide was without effect. However, at pH 5.8, whereas cathepsin E was virtually inactive by itself, it was restored to full activity (k
cat) by ATP and the non‐hydrolysable methylene‐ATP analogue. At still higher pH values, k
cat progressively diminished but significant levels of cathepsin E activity were readily detectable at pH 7.0. The specificity of this stabilisation effect was examined.
Electrostimulation‐induced increases in fatty acid‐binding protein and myoglobin in rat fast‐twitch muscle and comparison with tissue levels in heart
Authors: Kaufmann Margit , Simoneau Jean-Aimé , Veerkamp Jacques H. , Pette Dirk
Published: July 2002

Chronic stimulation of rat fast‐twitch muscle increased the content of both fatty acid‐binding protein (FABP) and myoglobin. The increases in FABP, which reached values close to that of cardiac muscle, exceeded those in myoglobin and those in citrate synthase and 3‐hydroxyacyl‐CoA dehydrogenase activities.
α1‐Adrenergic stimulation causes Mg2+ release from perfused rat liver
Authors: Jakob Andrea , Becker Johannes , Schöttli Gertrud , Fritzsch Günter
Published: July 2002

The possibility that Mg2+ mobilization is stimulated in perfused liver by α1‐adrenergic agonists was studied by measuring Mg2+ release in response to 0.5 and 20μM phenylephrine. During preperfusion exogenous Mg2+ was added to the medium to give 1.2 mM. 5 min before starting the addition of phenylephrine the infusion of exogenous Mg2+ was stopped. Mg2+ in the perfusate leaving the liver was measured by atomic absorption spectroscopy. Analysis of the Mg2+ decay curves with two exponential models indicated that phenylephrine caused dose‐dependent Mg2+ release from perfused rat livers.
Induction of prostacyclin receptor expression in human erythroleukemia cells
Authors: Murray Rosemary , Furci Lucinda , FitzGerald Garret A.
Published: July 2002

We have identified both high‐affinity (KD = 36±3 nM) and low‐affinity (KD = 2.1±0.8 μM) prostacyclin (PGI2)‐receptor sites on human erythroleukemia (HEL) cells using the radiolabelled prostacyclin analogue, [3H]iloprost. The addition of the phorbol ester, TPA, to the culture medium caused a 5–10‐fold increase in the number of both the low‐ and the high‐affinity sites, without any change in their affinity constants. Iloprost stimulated HEL cell membrane adenylate cyclase activity 5‐fold. This stimulation was potentiated in the presence of GTP, indicating a conventional PGI2 receptor‐Gs‐adenylate cyclase system. HEL cells represent a source of prostacyclin receptor mRNA which may be of value in expression cloning of this receptor.
O‐ and N‐glycosylation lead to different molecular mass forms of human monocyte interleukin‐6
Authors: Gross Volker , Andus Tilo , Castell José , Vom Berg Daniela , Heinrich Peter C. , Gerok Wolfgang
Published: July 2002

The biosynthesis and secretion of human interleukin‐6 (IL‐6) was studied in monocyte cultures stimulated with endotoxin. After labeling with [35S]methionine and immunoprecipitation with a specific antiserum one major (24 kDa) and four minor (27.5, 23.3, 22.5 and 21.8 kDa) molecular mass forms of IL‐6 could be found in the cells and media. Incubation of monocyte media with sialidase and subsequently with endo‐α‐N‐acetylgalactosaminidase, which cleaves Ga1(β1‐3)Gal‐NAc from serine or threonine, led to the formation of only two forms of IL‐6 with apparent molecular masses of 25 and 21.8 kDa. The latter had an electrophoretic mobility indistinguishable from that of 125I‐labeled recombinant human IL‐6. The results suggest that human monocyte IL‐6 carries O‐glycosidically bound carbohydrates with a Ga1(β1‐3)Gal‐NAc core to which only sialic acid is bound. Differences in O‐glycosylation are the major cause for the molecular heterogeneity of IL‐6. A small part of IL‐6 (27.5 kDa form) is in addition N‐glycosylated. Incubation of monocytes with tunicamycin and 1‐deoxymynnojirimycin and treatment of IL‐6 with endoglucosaminidase H suggested that the 27.5 kDa form of IL‐6 carries at least one N‐linked complex‐type oligosaccharide chain.
Platelet‐activating factor increases inositol phosphate production and cytosolic free Ca2+ concentrations in cultured rat Kupffer cells
Authors: Fisher Rory A. , Sharma Ram V. , Bhalla Ramesh C.
Published: July 2002

Platelet‐activating factor (PAF) stimulates glycogenolysis in perfused livers but not in isolated hepatocytes [(1984) J. Biol. Chem. 259, 8685–8688]. PAF‐induced glycogenolysis in liver is associated closely with a pronounced constriction of the hepatic vasculature [(1986) J. Biol. Chem. 261, 644–649]. These and other observations suggest that PAF stimulates glycogenolysis in liver indirectly by interactions with cells other than hepatocytes. We have evaluated effects of PAF on hepatic Kupffer cells, which regulate flow through the hepatic sinusoids. Application of PAF to [3H]inositol‐labeled Kupffer cells produced dose‐dependent increases in [3H]inositol phosphates with an EC50 value of 4 × 10−10 M. Increasesin inositol phosphate production in response to PAF were inhibited by a specific PAF receptor antagonist, SRI 63‐675 (2 × 10−7 M), and stimulus of protein kinase C, phorbol 12‐myristate 13‐acetate (1 × 10−7 M). Measurements of cytosolic free Ca2+ concentrations ([Ca2+]i) in single Kupffer cells loaded with Fura‐2 demonstrated that application of PAF (2 × 10−9 M) resulted in significant increases in [Ca2+]i. These observations lead us to propose that interactions of PAF with Kupffer cells may result in the hemodynamic and metabolic responses to PAF in liver.
Identification of psbI and psbL gene products in cyanobacterial photosystem II reaction center preparation
Authors: Ikeuchi Masahiko , Koike Hiroyuki , Inoue Yorinao
Published: July 2002

Photosystem II (PS II) reaction center complex of higher plants was revealed to contain a new low‐molecular‐mass component of 4.8 kDa encoded by a chloroplast gene, psbI [(1988) FEBS Lett. 241, 99–104], in addition to D1, D2 and two subunits of cytochrome b‐559. Here we isolated a PS II reaction center complex from the thermophilic cyanobacterium, Synechococcus vulcanus, by column chromatography using Triton X‐100 and urea, and attempted to identify the same gene product. SDS gel electrophoresis of the cyanobacterial reaction center complex resolved three protein bands of 8, 5.4 and 5 kDa in addition to D1, D2 and a partially remaining 47 kDa protein. N‐terminal sequencing revealed that the 5 kDa band consisted of two components: one, having an open N‐terminus, was homologous to the psbL gene product of higher plants, and the other, having a blocked N‐terminus, was homologous to the psbI gene product of higher plants. The 8 kDa and 5.4 kDa were the large and small subunits of cytochrome b‐559, respectively. Based on these results, we conclude that the subunit structure of cyanobacterial PS II reaction center complex is essentially the same as that of higher plants except that the cyanobacterial psbL product is more tightly associated with the reaction center.
Identification of a small intracellular region of the muscarinic m3 receptor as a determinant of selective coupling to PI turnover
Authors: Wess Jürgen , Brann Mark R. , Bonner Tom I.
Published: July 2002

Molecular cloning studies have demonstrated the existence of five different muscarinic receptors (ml‐m5). While ml, m3 and m5 strongly couple to stimulation of phosphoinositide (PI) hydrolysis, m2 and m4 are more efficiently linked to inhibition of adenylate cyclase. The sequences of m1–m5 ave a short segment at the N‐terminal portion of the putative third cytoplasmic loop (i3) which is highly conserved among ml, m3 and m5, but different from the sequence which is well conserved among m2 and m4. To study the role of this region in conferring coupling selectivity, we constructed cDNAs encoding chimeric receptors. Transient expression of these receptor hybrids in COS‐7 cells showed that a 17 amino acid segment at the N‐terminal portion of i3 is a major determinant of how efficiently the different muscarinic receptors are coupled to PI hydrolysis.
Correlation of some published amino acid sequences for photosystem I polypeptides to a 17 kDa LHCI pigment‐protein and to subunits III and IV of the core complex
Authors: Anandan Shivanthi , Vainstein Alexander , Thornber J.Philip
Published: July 2002

Photosynthesis; Photosystem I subunit; Light‐harvesting complex; Protein sequence; Pigment protein