Sequestration of muscarinic acetylcholine receptor m2 subtypes. Facilitation by G protein-coupled receptor kinase (GRK2) and attenuation by a dominant-negative mutant of GRK2

Sequestration of m2 receptors (muscarinic acetylcholine receptor m2 subtypes), which was assessed as loss of N-[3H]methylscopolamine ([3H]NMS) binding activity from the cell surface, was examined in COS 7 and BHK-21 cells that had been transfected with expression vectors encoding the m2 receptor and, independently, vectors encoding a G protein-coupled receptor kinase (GRK2) (beta-adrenergic receptor kinase 1) or a GRK2 dominant-negative mutant (DN-GRK2). The sequestration of m2 receptors became apparent when the cells were treated with 10(-5) M or higher concentrations of carbamylcholine. In this case, approximately 40% or 20-25% of the [3H]NMS binding sites on COS 7 or BHK-21 cells, respectively, were sequestered with a half-life of 15-25 min. In cells in which GRK2 was also expressed, the sequestration became apparent in the presence of 10(-7) M carbamylcholine. Approximately 40% of the [3H]NMS binding sites on both COS 7 and BHK-21 cells were sequestered in the presence of 10(-6) M or higher concentrations of carbamylcholine. When DN-GRK2 was expressed in COS 7 cells, the proportion of [3H]NMS binding sites sequestered in the presence of 10(-5) M or higher concentrations of carbamylcholine was reduced to 20-30%. These results indicate that the phosphorylation of m2 receptors by GRK2 facilitates their sequestration. These results are in contrast with the absence of a correlation between sequestration and the phosphorylation of beta-adrenergic receptors by the GRK2 and suggests that the consequences of phosphorylation by GRK2 are different for different receptors.     

Our new fragile world doesn't consist of developed and developing countries only

Health conditions have improved worldwide; therefore, the division into developed and developing countries no longer holds true. International organizations tend to divide countries into three groups. An increasing number of people are born in middle income countries where health conditions continue to improve. However, in a number of the least developed countries, mortality is on the rise in a spiral of economic stagnation, environmental problems, social misery, and ethnic/civil conflicts. This requires the medical assistance of international agencies from abroad. Since the 1960s there has been a drastic decline of child mortality in developing countries, especially in Asia. 1 billion people live in countries with child mortality under 20/1000, almost 3 billion live in countries with child mortality ranging 20-100/1000, and over 1 billion live in the least developed countries with child mortality over 100/1000. Unicef divides countries into developed, developing, and least developed countries, while the World Bank groups them as high-, medium-, and low-income countries. Thailand's child mortality is the same as that of Russia, while Cuba has a lower rate than Washington, D.C. On the other hand, Singapore is a developed high-income country with one of the world's healthiest populations. Stagnation and conflicts in the former socialist countries mean that many Asian and Latin American countries have better health status than some parts of Europe. Despite Africa's high mortality, its population growth is the highest in the world: in 20 years its population has doubled. Economic stagnation and the debt burden in many of these countries has resulted in ethnic conflicts and the collapse of social institutions: Somalia, southern Sudan, Rwanda, Liberia, Angola, perhaps Zaire, and Mozambique. The organization Physicians Beyond Borders is an example of extending humanitarian help and combatting social collapse in the least developed countries.     

Interleukin-6 cDNA transfected Lewis lung carcinoma cells show unaltered net tumour growth rate but cause weight loss and shortened survival in syngeneic mice

HuIL-6 cDNA, cloned into a neomycin resistant conferring expression vector, BMGNeo, was transfected into Lewis Lung Carcinoma (LLC) cells. LLC cells (5 x 10(6) ml-1) transfected with IL-6 cDNA (LLC-IL6) secreted IL-6 into the culture supernatant at a concentration of 9.9 ng ml-1 within 48 h. When 1,000,000 of untransfected LLC, BMGNeo vector transfected LLC (LLC-Neo) or LLC-IL6 cells were transplanted into C57BL/6 mice subcutaneously, the mean +/- s.d. of survival times of these mice were 33.3 +/- 9.7, 34.3 +/- 7.1 and 17.0 +/- 3.1 days, respectively. The survival time of LLC-IL6 cells transplanted mice was significantly shorter than that of LLC (P < 0.01) or LLC-Neo (P < 0.01) cells transplanted mice without a measurable difference of tumour size. Plasma concentration of IL-6 steadily increased in LLC-IL6 transplanted mice. Body weight and serum albumin were significantly lower in LLC-IL6 transplanted mice than in LLC transplanted mice. Mouse IL-1 alpha and mouse TNF-alpha were not detected in the plasma of LLC-IL6 transplanted mice. These data suggested that secretion of IL-6 from LLC cells was unable to alter net tumour growth rate but rather caused a state similar to cachexia without detectable increase of IL-1 alpha and TNF-alpha in the plasma. This state may be responsible for the shortened survival of LLC-IL6 tumour-bearing mice.     

A case of venous aneurysm

A 70-year-old woman was referred to our hospital with a complaint of a soft mass at the right elbow. Venography demonstrated that the mass was connected with vein. This mass was surgically removed and histologically a marked reduction of both elastic fibers and the adventitia were observed in the aneurysmal wall. Venous aneurysm (VA) is different form varicose veins by many features. It was considered that congenital fragility of the venous walls was associated with the onset of VA. We classified VA into type-I VA (cystic ectasia) and type II (fusiform ectasia).     

Morphological and functional polymorphism within clonal thyroid nodules

Thirty-nine thyroid nodules, removed because of recent growth, were analyzed morphologically by serial histological sections for the classical histomorphological hallmarks of follicular cell replication and for immunohistochemically demonstrable overexpression of the growth-associated ras-gene product p21ras. Clonal analysis was performed using the highly informative probe M27 beta that detects polymorphisms on the locus DXS255 of the X-chromosome. Twenty-four nodules were of clonal and 15 nodules were of poly-clonal origin. Only 3 out of the 24 clonal nodules were histomorphologically uniform. In all others, the structural hallmarks of active growth and the P21ras growth-marker expression were remarkably heterogeneous throughout the tumors. There were no histomorphological characteristics distinguishing these clonal tumors from polyclonal nodules. Even if a clonal thyroid tumor may be originally homogeneous in respect to the parameters studied here, mechanisms must exist that create wide heterogeneity of growth and of morphogenetic potential among the individual follicular cells during further expansion of the nodule. Thus, clonal nodules are much more common in nodular goiters than hitherto assumed on grounds of the classical morphological criteria. The diagnosis of a true monoclonal nodule can no longer rely on morphological and functional criteria alone but requires molecular or cytogenetic analysis of clonality.     

Drug therapy of hemiballism. Review of the literature and personal observations

Hemiballism is a rare hyperkinetic disorder which is characterized by irregular vigorous rotatory movements of the limbs of one side of the body. The disabling and exhausting movements with their high risk of severe injuries require an acute drug treatment. However, there are no common guidelines for the treatment of this disorder. Therefore we reviewed the available literature to establish a concept for a rational symptomatic drug treatment. Furthermore, own observations in 6 patients with hemiballism of different origins are reported.     

Solar/UV-induced photocatalytic degradation of three commercial textile dyes.

The photocatalytic degradation of three commercial textile dyes with different structure has been investigated using TiO(2) (Degussa P25) photocatalyst in aqueous solution under solar irradiation. Experiments were conducted to optimise various parameters viz. amount of catalyst, concentration of dye, pH and solar light intensity. Degradation of all the dyes were examined by using chemical oxygen demand (COD) method. The degradation efficiency of the three dyes is as follows: Reactive Yellow 17(RY17) > Reactive Red 2(RR2) > Reactive Blue 4 (RB4), respectively. The experimental results indicate that TiO(2) (Degussa P25) is the best catalyst in comparison with other commercial photocatalysts such as, TiO(2) (Merck), ZnO, ZrO(2), WO(3) and CdS. Though the UV irradiation can efficiently degrade the dyes, naturally abundant solar irradiation is also very effective in the mineralisation of dyes. The comparison between thin-film coating and aqueous slurry method reveals that slurry method is more efficient than coating but the problems of leaching and the requirement of separation can be avoided by using coating technique. These observations indicate that all the three dyes could be degraded completely at different time intervals. Hence, it may be a viable technique for the safe disposal of textile wastewater into the water streams.     

An eight‐node hybrid‐stress solid‐shell element for geometric non‐linear analysis of elastic shells

This paper presents eight‐node solid‐shell elements for geometric non‐linear analysis of elastic shells. To subdue shear, trapezoidal and thickness locking, the assumed natural strain method and an ad hoc modified generalized laminate stiffness matrix are employed. A selectively reduced integrated element is formulated with its membrane and bending shear strain components taken to be constant and equal to the ones evaluated at the element centroid. With the generalized stresses arising from the modified generalized laminate stiffness matrix assumed to be independent from the ones obtained from the displacement, an extended Hellinger–Reissner functional can be derived. By choosing the assumed generalized stresses similar to the assumed stresses of a previous solid element, a hybrid‐stress solid‐shell element is formulated. Commonly employed geometric non‐linear homogeneous and laminated shell problems are attempted and our results are close to those of other state‐of‐the‐art elements. Moreover, the hybrid‐stress element converges more readily than the selectively reduced integrated element in all benchmark problems. Copyright © 2002 John Wiley & Sons, Ltd.     

Simple concepts for ion source improvement

We report on improvements in the overall intensity of a sputter ion source that evolved originally from an NEC MCSNICS. Beam output increases benefit both AMS measurements and nuclear physics experiments using low natural abundance beams. In particular, minor changes in source geometry suggested by a combination of electrostatic calculations and simple design principles have yielded increases in extracted negative ion intensity of nearly a factor of 4.     

Creating and Justifying Research and Development Value: Scope,Scale, Skill and Social Networking of R&D

In this paper we describe a framework for analysing the creation and justification of Research & Development. The 4S framework is developed for analysing the scope, scale, skills and social network aspects of Research & Development value. The framework is based on social system theory, a process contingency model, and recent Research & Development metrics. We present a first empirical assessment based on a workshop using the 4S framework for leveraging Research & Development. Results that assist in the assessment of value creation utilising R & D within networks are very relevant in high tech industries. The multi–dimensional process approach of this framework seems promising for understanding and managing R&D value creation, but needs further operationalisation. Case studies are described and a Dutch network on leveraging R&D has been initiated.