bluuebunny/crossref_metadata_2025 huggingface dataset:
In situdetection of activated Bruton’s tyrosine kinase in the Ig signaling complex by phosphopeptide-specific monoclonal antibodies
Authors: Nisitani Sazuku , Kato Roberta M. , Rawlings David J. , Witte Owen N. , Wahl Matthew I.
Published: July 2002

Bruton’s tyrosine kinase (Btk) is a critical transducer of signals originating from the B cell antigen receptor (BCR). Dosage, sequential phosphorylation, and protein interactions are interdependent mechanisms influencing Btk function. Phosphopeptide-specific mAbs recognizing two distinct phosphotyrosine modifications were used to quantify Btk activation by immunofluorescent techniques during B cell stimulation. In a population of cultured B cells stimulated by BCR crosslinking and analyzed by flow cytometry, transient phosphorylation of the regulatory Btk tyrosine residues (551Y and 223Y) was detected. The kinetics of phosphorylation of the residues were temporally distinct. Tyrosine 551, a transactivating substrate site for Src-family kinases, was maximally phosphorylated within ≈30 seconds of stimulation as monitored by flow cytometry. Tyrosine 223, an autophosphorylation site within the SH3 domain, was maximally phosphorylated at ≈5 minutes. Btk returned to a low tyrosine phosphorylation level within 30 minutes, despite persistent elevation of global tyrosine phosphorylation. Colocalization of activated Btk molecules with the crosslinked BCR signaling complex was observed to coincide with the period of maximal Btk tyrosine phosphorylation when stimulated B cells were analyzed with confocal microscopy. The results of thesein situtemporal and spatial analyses imply that Btk signaling occurs in the region of the Ig receptor signaling complex, suggesting a similar location for downstream targets of its activity.
Site-specific integration mediated by a hybrid adenovirus/adeno-associated virus vector
Authors: Recchia Alessandra , Parks Robin J. , Lamartina Stefania , Toniatti Carlo , Pieroni Luisa , Palombo Fabio , Ciliberto Gennaro , Graham Frank L. , Cortese Riccardo , La Monica Nicola , Colloca Stefano
Published: July 2002

Adenovirus (Ad) and adeno-associated virus (AAV) have attractive and complementary properties that can be exploited for gene transfer purposes. Ad vectors are probably the most efficient vehicles to deliver foreign genes both
in vitro
and
in vivo
. AAV exhibits the unique ability to establish latency by efficiently integrating at a specific locus of human chromosome 19 (AAVS1). Two viral elements are necessary for the integration at AAVS1: Rep68/78 and the inverted terminal repeats (AAV-ITRs). In this study, we report the development of two helper-dependent adenoviral (HD) vectors, one carrying the Rep78 gene, the other an AAV-ITR-flanked transgene. Although Rep proteins have been demonstrated to interfere with Ad replication, HD Rep78 vector was successfully amplified on serial passages in 293CRE4 cells with a yield of 50–100 transducing units per cell. DNA integration at the AAVS1 site also was demonstrated in hepatoma cells coinfected with the HD-expressing Rep78 and with the second HD vector carrying a transgene flanked by AAV-ITRs. The high transduction efficiency, large cloning capacity, and high titer of the HD, combined with the site-specific integration machinery provided by AAV-derived components, make the Ad/AAV hybrid viruses a promising vehicle for gene therapy.
Molecular evidence for a relationship between LINE-1 elements and X chromosome inactivation: The Lyon repeat hypothesis
Authors: Bailey Jeffrey A. , Carrel Laura , Chakravarti Aravinda , Eichler Evan E.
Published: July 2002

X inactivation is a chromosome-specific form of genetic regulation in which thousands of genes on one homologue become silenced early in female embryogenesis. Although many aspects of X inactivation are now understood, the spread of the X inactivation signal along the entire length of the chromosome remains enigmatic. Extending the Gartler–Riggs model [Gartler, S. M. & Riggs, A. D. (1983)
Annu. Rev. Genet.
17, 155–190], Lyon recently proposed [Lyon, M. F. (1998)
Cytogenet. Cell Genet.
80, 133–137] that a nonrandom organization of long interspersed element (LINE) repetitive sequences on the X chromosome might be responsible for its facultative heterochromatization. In this paper, we present data indicating that the LINE-1 (L1) composition of the human X chromosome is fundamentally distinct from that of human autosomes. The X chromosome is enriched 2-fold for L1 repetitive elements, with the greatest enrichment observed for a restricted subset of LINE-1 elements that were active <100 million years ago. Regional analysis of the X chromosome reveals that the most significant clustering of these elements is in Xq13–Xq21 (the center of X inactivation). Genomic segments harboring genes that escape inactivation are significantly reduced in L1 content compared with X chromosome segments containing genes subject to X inactivation, providing further support for the association between X inactivation and L1 content. These nonrandom properties of L1 distribution on the X chromosome provide strong evidence that L1 elements may serve as DNA signals to propagate X inactivation along the chromosome.
Nonreceptor tyrosine protein kinase pp60c-src in spatial learning: Synapse-specific changes in its gene expression, tyrosine phosphorylation, and protein–protein interactions
Authors: Zhao Weiqin , Cavallaro Sebastiano , Gusev Pavel , Alkon Daniel L.
Published: July 2002

c-src is a nonreceptor tyrosine protein kinase that is highly concentrated in synaptic regions, including synaptic vesicles and growth cones. Here, we report that the mRNA signal of pp60c-src is widely distributed in the rat brain with particularly high concentrations in the hippocampus. After spatial maze learning, up-regulation of c
-src
mRNA was observed in the CA3 region of the hippocampus, which was accompanied by increases in pp60c-src protein in hippocampal synaptosomal preparations. Training also triggered an increase in c-src protein tyrosine kinase activity that was correlated with its tyrosine dephosphorylation in the synaptic membrane fraction. After training, pp60c-src from hippocampus showed enhanced interactions with synaptic proteins such as synapsin I, synaptophysin, and the type 2
N
-methyl-
d
-aspartate receptor, as well as the cytoskeletal protein actin. The association of pp60c-src with insulin receptor in the synaptic membrane fraction, however, was temporally decreased after training. Furthermore,
in vitro
results showed that Ca
2+
and protein kinase C might be involved in the regulation of protein–protein interactions of pp60c-src. These results suggest, therefore, that pp60c-src participates in the regulation of hippocampal synaptic activity during learning and memory.
The role of antimicrobial peptides in animal defenses
Authors: Hancock Robert E. W. , Scott Monisha G.
Published: July 2002

It is becoming clear that the cationic antimicrobial peptides are an important component of the innate defenses of all species of life. Such peptides can be constitutively expressed or induced by bacteria or their products. The best peptides have good activities vs. a broad range of bacterial strains, including antibiotic-resistant isolates. They kill very rapidly, do not easily select resistant mutants, are synergistic with conventional antibiotics, other peptides, and lysozyme, and are able to kill bacteria in animal models. It is known that bacterial infections, especially when treated with antibiotics, can lead to the release of bacterial products such as lipopolysaccharide (LPS) and lipoteichoic acid, resulting in potentially lethal sepsis. In contrast to antibiotics, the peptides actually prevent cytokine induction by bacterial products in tissue culture and human blood, and they block the onset of sepsis in mouse models of endotoxemia. Consistent with this, transcriptional gene array experiments using a macrophage cell line demonstrated that a model peptide, CEMA, blocks the expression of many genes whose transcription was induced by LPS. The peptides do this in part by blocking LPS interaction with the serum protein LBP. In addition, CEMA itself has a direct effect on macrophage gene expression. Because cationic antimicrobial peptides are induced by LPS and are able to dampen the septic response of animal cells to LPS, we propose that, in addition to their role in direct and lysozyme-assisted killing of microbes, they have a role in feedback regulation of cytokine responses. We are currently developing variant peptides as therapeutics against antibiotic-resistant infections.
The RING-H2 finger protein APC11 and the E2 enzyme UBC4 are sufficient to ubiquitinate substrates of the anaphase-promoting complex
Authors: Gmachl Michael , Gieffers Christian , Podtelejnikov Alexandre V. , Mann Matthias , Peters Jan-Michael
Published: July 2002

The anaphase-promoting complex (APC) is a cell cycle-regulated ubiquitin-protein ligase that targets cyclin B, securin and other destruction box containing proteins for proteolysis. Nine APC subunits have been identified in vertebrates and eleven in yeast, but for none of them it is known how they contribute to the catalysis of ubiquitination reactions. Here we report the mass spectrometric identification of CDC26 and of the RING-H2 finger protein APC11 in the human APC. We have expressed these proteins and several other APC subunits in
Escherichia coli
and have tested their activities
in vitro
. We find that APC11 alone is sufficient to allow the synthesis of multiubiquitin chains in the presence of E1 and UBC4. These multiubiquitin chains are partly unanchored and partly bound to APC11 itself. APC11 and UBC4 are also able to ubiquitinate securin and cyclin B, but these reactions show a decreased dependency on the destruction box. The integrity of the putative zinc binding RING-H2 finger is required for the ability of APC11 to support ubiquitination reactions. These results suggest that APC11 and UBC4 catalyze the formation of isopeptide bonds in APC-mediated ubiquitination reactions.
Role of C-terminal domains of the G protein β subunit in the activation of effectors
Authors: Myung Chang-Seon , Garrison James C.
Published: July 2002

The prenyl group on the G protein γ subunit is an important determinant of protein–protein interactions between the βγ dimer and its targets, such as α subunits, receptors, and effectors. In an effort to identify domains of the β subunit important for the activation of effectors, we have prepared two types of mutants, one set in the domain suggested to form a hydrophobic prenyl-binding pocket for the γ subunit's prenyl group (prenyl pocket mutants) and the other set in a domain between Gly
306
and Gly
319
in the β propeller, which undergoes a conformational change when the dimer binds to phosducin (conformational change mutants). Recombinant baculoviruses for each set of mutants were prepared, and the nine mutant β subunits were overexpressed with either the γ
2
subunit (modified with geranylgeranyl) or the γ
2-L71S
subunit (γ
2
with altered
CAAX
sequence and modified with farnesyl). The purified dimers were tested for their ability to couple Gα
i1
to the A1 adenosine receptor and to activate phospholipase C-β or type II adenylyl cyclase. All dimers containing mutant β subunits were indistinguishable from wild-type β
1
γ
2
or β
1
γ
2-L71S
in coupling the receptor to Gα
i1
. The prenyl pocket mutants expressed with γ
2
were 10-fold less potent in activating phospholipase C-β and adenylyl cyclase than β
1
γ
2
and had similar activities to β
1
γ
2-L71S
. The conformational change mutants caused a 15- to 23-fold decrease in EC
50
values for activation of these two effectors. Overall, the results suggest that the sites in Gβ identified by these mutants are very important in the activation of effectors. Furthermore, the nature of the prenyl group on Gγ may play an important role in the conformational change leading to the activity of Gβγ on effectors.
Etiology of interepidemic periods of mosquito-borne disease
Authors: Hay Simon I. , Myers Monica F. , Burke Donald S. , Vaughn David W. , Endy Timothy , Ananda Nisalak , Shanks G. Dennis , Snow Robert W. , Rogers David J.
Published: July 2002

Dengue viruses and malaria protozoa are of increasing global concern in public health. The diseases caused by these pathogens often show regular seasonal patterns in incidence because of the sensitivity of their mosquito vectors to climate. Between years in endemic areas, however, there can be further significant variation in case numbers for which public health systems are generally unprepared. There is an acute need for reliable predictions of within-year and between-year epidemic events. The prerequisite for developing any system of early warning is a detailed understanding of the factors involved in epidemic genesis. In this report we discuss the potential causes of the interepidemic periods in dengue hemorrhagic fever in Bangkok and ofPlasmodium falciparummalaria in a highland area of western Kenya. The alternative causes are distinguished by a retrospective analysis of two unique and contemporaneous 33-year time series of epidemiological and associated meteorological data recorded at these two sites. We conclude that intrinsic population dynamics offer the most parsimonious explanation for the observed interepidemic periods of disease in these locations.
The putative tumor suppressors EXT1 and EXT2 form a stable complex that accumulates in the Golgi apparatus and catalyzes the synthesis of heparan sulfate
Authors: McCormick Craig , Duncan Gillian , Goutsos K. Tina , Tufaro Frank
Published: July 2002

Hereditary multiple exostoses, a dominantly inherited genetic disorder characterized by multiple cartilaginous tumors, is caused by mutations in members of the
EXT
gene family,
EXT1
or
EXT2
. The proteins encoded by these genes, EXT1 and EXT2, are endoplasmic reticulum-localized type II transmembrane glycoproteins that possess or are tightly associated with glycosyltransferase activities involved in the polymerization of heparan sulfate. Here, by testing a cell line with a specific defect in EXT1 in
in vivo
and
in vitro
assays, we show that EXT2 does not harbor significant glycosyltransferase activity in the absence of EXT1. Instead, it appears that EXT1 and EXT2 form a hetero-oligomeric complex
in vivo
that leads to the accumulation of both proteins in the Golgi apparatus. Remarkably, the Golgi-localized EXT1/EXT2 complex possesses substantially higher glycosyltransferase activity than EXT1 or EXT2 alone, which suggests that the complex represents the biologically relevant form of the enzyme(s). These findings provide a rationale to explain how inherited mutations in either of the two
EXT
genes can cause loss of activity, resulting in hereditary multiple exostoses.
Enzymatic reduction of disulfide bonds in lysosomes: Characterization of a Gamma-interferon-inducible lysosomal thiol reductase (GILT)
Authors: Arunachalam Balasubramanian , Phan Uyen T. , Geuze Hans J. , Cresswell Peter
Published: July 2002

Proteins internalized into the endocytic pathway are usually degraded. Efficient proteolysis requires denaturation, induced by acidic conditions within lysosomes, and reduction of inter- and intrachain disulfide bonds. Cytosolic reduction is mediated enzymatically by thioredoxin, but the mechanism of lysosomal reduction is unknown. We describe here a lysosomal thiol reductase optimally active at low pH and capable of catalyzing disulfide bond reduction bothin vivoandin vitro. The active site, determined by mutagenesis, consists of a pair of cysteine residues separated by two amino acids, similar to other enzymes of the thioredoxin family. The enzyme is a soluble glycoprotein that is synthesized as a precursor. After delivery into the endosomal/lysosomal system by the mannose 6-phosphate receptor, N- and C-terminal prosequences are removed. The enzyme is expressed constitutively in antigen-presenting cells and induced by IFN-γ in other cell types, suggesting a potentially important role in antigen processing.
Characteristic length scale of the intermediate structure in zero-pressure-gradient boundary layer flow
Authors: Barenblatt G. I. , Chorin A. J. , Prostokishin V. M.
Published: July 2002

In a turbulent boundary layer over a smooth flat plate with zero pressure gradient, the intermediate structure between the viscous sublayer and the free stream consists of two layers: one adjacent to the viscous sublayer and one adjacent to the free stream. When the level of turbulence in the free stream is low, the boundary between the two layers is sharp, and both have a self-similar structure described by Reynolds-number-dependent scaling (power) laws. This structure introduces two length scales: one—the wall-region thickness—determined by the sharp boundary between the two intermediate layers and the second determined by the condition that the velocity distribution in the first intermediate layer be the one common to all wall-bounded flows and in particular coincide with the scaling law previously determined for pipe flows. Using recent experimental data, we determine both these length scales and show that they are close. Our results disagree with the classical model of the “wake region.”
Cultural Difference in Hand Preference: Evidence from India and Japan
Authors: Mandal Manas K. , Ida Yukihide , Harizuka Susumu , Upadhaya Neelam
Published: July 2002

Indian (N = 400) and Japanese (N = 502) adult subjects were examined for their hand preference on a 7‐point scale (1 = left always, 7 = right always) of the 32‐item Waterloo Handedness Questionnaire (Steenhuis & Bryden, 1989). Factor analyses of the data yielded a two‐factorial structure of hand preference: skilled and unskilled. Interaction of Culture × Hand preference indicated that Indians had more right‐hand preference for unskilled activities whereas Japanese subjects had more right‐hand preference for skilled activities. Further analyses revealed that the frequency of middle category responses was more common in Japanese subjects. Indian subjects were found to give more extreme responses for either right‐ or left‐hand preference. Findings are discussed in the light of cultural training given to the individuals in these two societies, which are essentially collective in nature.
“The Mythical Jew”: Antisemitism, Intellectuals, and Democracy in Post-Communist Romania
Authors: Rosenthal Denise
Published: July 2002

A mentally healthy human being can go insane if suddenly diagnosed with leprosy. Eugen Ionescu finds out that even the “Ionescu” name, an indisputable Romanian father, and the fact of being born Christian can do nothing, nothing, nothing to cover the curse of having Jewish blood in his veins. With resignation and sometimes with I don't know what sad and discouraged pride, we got used to this dear leprosy a long time ago.

With these words, the Romanian–Jewish writer Mihail Sebastian expresses within his private diary some of the darkest moments of a World War II “transfigured” Romania, populated as they are by the gothic characters of legionaries, Nazis, and antisemitism. His death soon followed in 1945, when Romania was at the threshold of fascism and communism. However, with the discovery and the subsequent publishing of Sebastian's diary in 1996, and following 50 years of communist mystification of the Jewish Holocaust, the entire chaotic war atmosphere with the fascist affections of the Romanian intellectual elite was once again brought to light with all the flavor and scent of the dark past. In this entry from Sebastian's diary he speaks of his friend, Eugen Ionescu who, born of a French-related mother and a Romanian father, was living in Bucharest at that time. He would later become known to the world as Eugène Ionesco, the famous French playwright and author of the well-known playsThe Bald SopranoandThe Rhinoceros.The above quote from Sebastian's journal, predating the international fame of Ionesco, but already marking the end of Sebastian's career under fascism, remains a traumatizing testimony of the Jewish Kafkian torment as “guilt,” a deeply claustrophobic identity that many Eastern European Jewish intellectuals have learned to internalize. Beyond this symbolism, the publishing of Sebastian's diary in Romania unintentionally challenged an existent post-communist tendency of legitimizing inter-war fascist personalities within the framework of a general lack of knowledge about the Jewish Holocaust in both the communist and post-communist periods.
Early School Leaving and the Cultural Geography of High Schools [1]
Authors: Smyth John , Hattam Robert
Published: July 2002

Abstract Early school leaving is one of the most protracted educational problems around the world, but one of the least understood. Central to the issue itself, is the failure by the educational policy community to have ways of adequately ‘naming’ the problem. The study reported in this paper examines early school leaving from the position of 209 young Australians who had left school or who were at imminent risk of doing so. While acknowledging the considerable complexity of the decision making processes that lie behind this problem, this article provides a tentative theorising that traverses aspects of what we call the ‘cultural geography of the high school’ as a partial explanation of what is occurring. The question being pursued was how the culture of the school contributed to or interfered with early school leaving.
Naturally Occurring and Experimental Diabetes in Cynomolgus Monkeys: A Comparison of Carbohydrate and Lipid Metabolism and Islet Pathology
Authors: Wagner Janice D. , Cline J. Mark , Shadoan Melanie K. , Bullock Bill C. , Rankin Samuel E. , Cefalu William T.
Published: July 2002

Diabetes is a major health problem of increasing incidence in the United States. Diabetes research has been limited by lack of availability of good animal models, particularly for the study of comorbidities associated with diabetes. We investigated the use of cynomolgus monkeys as an animal model of both type 1 and type 2 diabetes and compared these naturally occurring diseases with streptozotocin-induced diabetes. Both type 1 diabetics and streptozotocin-induced diabetics present with sudden onset of hyperglycemia and are ketosis prone without exogenous insulin. Type 2 diabetics can have a very long period of moderate hyperglycemia and hypertriglyceridemia and only require exogenous insulin therapy if pancreatic islet reserves are depleted. Type 2 diabetes is preceded by a relatively long period of insulin resistance that is associated with obesity and dyslipidemia. As insulin resistance progresses, islet size and insulin content increases initially. However, with sustained periods of insulin resistance, islet amyloid polypeptide (IAPP) is deposited in islets and can replace normal islet architecture, resulting in an insulin-deficient state. Appearance of IAPP also occurs in human type 2 diabetics but not in conventional rodent models. Unlike type 2 diabetes, neither type 1 nor streptozotocin-induced diabetes is associated with IAPP. Rather, islets can appear normal histologically, but have decreased insulin secretion and immunostaining. Further, the amount of insulin present in the islet is correlated with plasma insulin levels following glucose challenge. Studies are ongoing to determine the pathogenic changes associated with the progression of diabetes and to find novel drug treatments for diabetics.
Heroin‐related deaths in regional New South Wales, 1992–96
Authors: DARKE SHANE , ROSS JOANNE
Published: July 2002

The coronial files of all 188 heroin‐related fatalities that occurred in regional New South Wales between 1992 and 1996 were inspected. There was a significant increase in fatalities, rising from 23 deaths in 1992 to 53 during 1996. The regions in which the most deaths occurred were Wollongong/Illawarra (43 deaths), Newcastle/Hunter (35) and the far north coast (25). The mean age of cases was 31.5 years and 83% were male, and there were no significant trends in demographic characteristics of cases over the study period. The median blood morphine concentration of cases was 0.39 mg/l (range 0.05–4.5 mg/l). Alcohol was detected in 50% of cases and benzodiazepines in 29%. There were large regional variations in toxicology results, with median blood morphine concentrations ranging from 0.25mg/l among south coast cases to 0.56mg/l in mid‐western New South Wales. Compared to Sydney metropolitan cases, regional cases had a higher median blood morphine concentration, were less likely to have cocaine detected, were more likely to have died in a home environment and to have been born in Australia.
Attitudes and beliefs of doctors prescribing methadone to addicts in the Westfalen‐Lippe region of Germany
Authors: GERLACH RALF , CAPLEHORN JOHN R.M.
Published: July 2002

Doctors who prescribe methadone to heroin addicts in the Westfalen‐Lippe region of Germany were surveyed in 1996 to develop and use German versions of existing Australian and American scales. Two scales were developed from 247 responses using confirmatory factor analysis and Cronbach's alpha. The 10‐item abstinence orientation (AO) scale measures strength of commitment to abstinence‐orientated treatment policies, and the five‐item disapproval of drug use (DDU) scale measures disapproval of illicit drug use. Half the respondents supported and a quarter strongly supported abstinence‐orientated policies, while a quarter supported a punitive societal response to illicit drug use. Scores on the two scales were highly and positively correlated; the stronger the support for abstinence‐orientated policies, the greater the disapproval of illicit drug use. The doctors' median score on a test of knowledge of the risks and benefits of methadone maintenance was six of a possible 10. While allowing for DDU scores, support for abstinence‐orientated policies was stronger among the less well‐informed doctors. The attitudes and beliefs and lack of knowledge identified in the survey are a cause for concern. Previous work has shown there were similar problems among methadone prescribers and staff of methadone units in Sydney, Australia and New York City, USA.
Gradual Dose Taper Following Chronic Buprenorphine
Authors: Becker Ami B. , Strain Eric C. , Bigelow George E. , Stitzer Maxine L. , Johnson Rolley E.
Published: July 2002

This paper describes the time course of withdrawal and relapse in opioid‐dependent volunteers (n = 8) who completed a gradual outpatient buprenorphine dose taper (28 days). Compliance with treatment was very high, as evidenced by clinic attendance (96–100%). Urinalysis showed that 6 of the 8 volunteers had relapsed to opiates by the end of the dose taper, even though reports of withdrawal were generally low. Relapse may have been triggered by a desire to re‐experience the drug's positive subjective effects, craving, or low motivation to remain drug‐free. A longer taper combined with an expanded range of treatments may improve prognosis.
Reduced sensory and motor nerve conduction velocities in moderate drinkers
Authors: D'Amour Monique L. , Brissette Suzanne , Lavoie Joel , Butterworth Roger F.
Published: July 2002

The mechanism responsible for peripheral nerve dysfunction in chronic alcoholism has not been fully elucidated either in terms of its relationship to the quantity of alcohol consumed or to nutritional status. As part of a series of studies to address these issues, the effects of moderate drinking (60–90 g ethanol per day) or heavy drinking (> 100 g ethanol per day) on peripheral nervous function and thiamine status was measured in 73 patients admitted to a detoxification unit. Electromyographic evaluation revealed significant reductions in median and ulnar sensory and motor nerve conduction velocities in both moderate drinkers (n = 30) and heavy drinkers (n = 43) compared to age‐matched controls. Twelve moderate drinkers and 25 heavy drinkers manifested clinical neurological signs of peripheral neuropathy. Thiamine deficiency, as revealed by erythrocyte transketolase activation assay, was detected in two moderate drinkers and seven heavy drinkers but was not significantly correlated with electromyographic alterations with the exception of ulnar nerves. These findings provide evidence for significant early peripheral nerve dysfunction in moderate drinkers and a possible contributory role of thiamine deficiency to the ulnar nerve conduction deficits. Whether deficits in other water‐soluble vitamins or a direct neurotoxic effect of ethanol are implicated in alcoholic peripheral neuropathy awaits further studies.
Effects of oral ethanol on end‐tidal CO2 tension in patients misusing alcohol
Authors: WOLFF C. B. , GARDNER W. N. , PETERS T. J.
Published: July 2002

This study examines the effect of oral ethanol ingestion on PetCO2 and other respiratory variables in alcohol‐misusing subjects. Twelve patients were given a loading dose of alcohol (0.270 g/kg) followed by a second dose (0.135 g/kg) half an hour later, increasing blood alcohol to a mean of 82.0 ± 10.3 (SEM) mg/100 ml at 1 hour. Five patients were classified as hazardous drinkers with evidence of mild alcohol dependence but no toxicity (Severity of Alcohol Dependency (SADQ) score < 15/60; serum carbohydrate‐deficient transferrin < 6%). These patients showed a significant fall in mean end‐tidal PCO2 (PetCO2) on alcohol (2.08 ± 0.61 mm Hg; p = 0.027) with slightly increased ventilation (1.76 ± 0.94 1 min−1, p = 0.14); responses similar to those previously reported for normal subjects. Seven patients had evidence of moderate alcohol dependency and/or recent use (SADQ > 15/60; CDT > 6%). In these patients P et CO 2 did not fall after alcohol challenge (mean change + 0.61 mm Hg; p > 0.30). There were significant correlations between the changes in P et CO 2 and CDT (n = 12; rs= 0.66, p = 0.019) and SADQ (n = 11; rs= 0.688, p = 0.019). The results show that PetCO2 control is abnormal in the alcohol dependency syndrome.
Phagocytosis of the protozoon Tetrahymena pyriformis as an endpoint in the estimation of cocaine salt and cocaine freebase toxicity
Authors: STEFANIDOU M. , ALEVISOPOULOS G. , MARAVELIAS C. , LOUTSIDIS C. , KOUTSELINIS A.
Published: July 2002

Cells of the ciliated protozoon Tetrahymena pyriformis strain W, grown in a peptone‐yeast medium, usually contain many phagocytic vacuoles. The phagocytic activity of this protozoon was studied in vivo using heat‐inactivated yeast stained with carmine after exposing the cultures for 1 hour to different doses of cocaine hydrochloride or cocaine freebase (crack) (0.5, 1 or 2 mg/100 ml of protozoan culture).The number of vacuoles formed indicated the phagocytic activity. Cocaine hydrochloride and crack caused a decrease of the phagocytic activity of the protozoon (p < 0.05) when compared to the control cultures. Furthermore, the two chemical forms of cocaine, salt and free‐base respectively, caused quantitatively different effects on the phagocytic activity. Crack produced an extensive decrease in phagocytosis, compared to equal concentrations of cocaine hydrochloride. These results suggest a possible relationship between cocaine abuse and the suppression of phagocytosis that may contribute to the impairment of immunity in drug misusers.
Book and Media Reviews
Authors: COOK CHRIS
Published: July 2002

Book reviewed in this article:

Book reviews in this column will primar ily be of titles focusing completely, or in part, on biolog ical aspects of addiction. However, significant titles of general relevance to the addictions field will also be included, even if they are not “biological”, as will titles of general methodological and clinical relevance, even if they are not on “addictions”. Similar considerations will apply to other media (software, audio tapes and CDs, videos, etc). However, specific “addictions” software applications seem to be relatively uncommon and, as these items are rarely reviewed elsewhere, we will endeavour to include reviews of some of the older programmes that are still useful, as well as new titles that appear. I would appreciate suggestions of any items suitable for reviews, but especially software and other media of specific relevance to the addictions. Please contact: Professor Chris Cook, Kent Institute of Medicine and Health Sciences, University of Kent at Canterbur y, Kent CT2 7NR, UK. Email: c.c.h.cook@ukc.ac.uk

Problem Drinking 3rd Edition NICK HEATHER & IAN ROBERTSON

Preventing Drunk Driving E. R. SHORE & J. R. FERRARI (Eds)

IRMA Addictions ‐ cannabis and cognitive functioning NADIA SOLIWIJ

Opium and the People. Opiate use and drug control policy in nineteenth and early twentieth century England Revised edition VIRGINIA BERRIDGE

Oxidative Stress in Skeletal Muscle A. Z. REZNICK (Ed.)

Laboratory Methods for the Detection of Mutations and Polymorphisms in DNA GRAHAM R. TAYLOR (Ed.)

The Scientist as Consultant—building new career opportunities C. J. SINDERMANN & T. K. SAWYER

Software Atlas of the Human Brain (Deluxe Edition) JURGEN K. MAI, JOSEPH ASSHEUER & GEORGE PAXINOS

The Human Brain—an introduction to the human nervous system MARION HALL & DAVID ROBINSON
Active versus passive cleft‐type speech characteristics
Authors: Harding Anne , Grunwell Pamela
Published: July 2002

Cleft palate speech is generally described in terms of nasal resonance, nasal emission and compensatory articulations. A longitudinal study of children at different stages of surgical treatment revealed a distinction between passive and active cleft‐type speech characteristics whereby passive characteristics were thought to be the product of structural abnormality or dysfunction and active characteristics were specific articulatory gestures replacing intended consonants. Passive and active patterns of articulation are described and defined in the context of three longitudinal studies of subjects who were at various stages of two different surgical regimes: five bilateral cleft lip and palate (BCLP) subjects aged 1;6‐4;6, 12 mixed unilateral cleft lip and palate (UCLP) and BCLP subjects aged 4;6‐7;6 and nine mixed UCLP and BCLP subjects aged 9;0‐11;0. Reference is also made to data from 12 mixed cleft‐type subjects aged 13;0 who had been treated with different surgical timing regimes. Comparison is made between the incidence of active versus passive processes in relation to oral structure. At age 4;6 speech samples taken from BCLP subjects who had been treated with 1‐stage versus 2‐stage palate repair all evidenced both active and passive processes. The lack of differentiation in speech results irrespective of their current surgical status, i.e. completely repaired palates versus residual cleft of the hard palate, was unexpected. Cleft‐type processes in completely repaired subjects might be accounted for by the inevitable anterior defect following repair of a bilateral cleft. Older subjects with structural defects also evidenced more clefttype processes. The relevance of distinguishing between active and passive processes is underlined by consideration of the effects of structural changes following surgery. The effect of surgery on seven subjects' speech is discussed using the active/passive distinction. Active cleft‐type characteristics did not change as a direct result of surgery, whereas passive characteristics were largely eliminated following surgery. A specific distinction is made between active and passive nasal fricatives, with the implication that active nasal fricatives may not be affected by surgical intervention, whereas passive nasal fricatives may be eliminated by surgery. Accurate distinction between active and passive patterns of articulation may serve to identify those cleft‐type speech error patterns most likely to respond to surgical intervention. Indications from this study are that active cleft‐type characteristics require destabilization in a course of speech and lan guage therapy before the potential benefits of surgery can be properly assessed. An analytical protocol for the interpretation of speech samples is presented and some therapy strategies are proposed for active and passive processes.
Role of the ADP/ATP‐Antiporter in Fatty Acid‐Induced Uncoupling of Ca2+‐Loaded Rat Liver Mitochondria
Authors: Bodrova M. E. , Dedukhova V. I. , Samartsev V. N. , Mokhova E. N.
Published: July 2002

We show that Ca2+
Scavenger Receptors and Modified Lipoproteins: Fatal Attractions?
Authors: Zingg Jean‐Marc , Ricciarelli Roberta , Azzi Angelo
Published: July 2002

Lipoproteins modified by oxidation, glycation, alkylation, and nitration are generated by oxidative stress during inflammation, diabetes, and inadequate supply of dietary antioxidants. A family of genes, the scavenger receptors, recognizes and internalizes modified lipoproteins, making them susceptible to degradation. Clearance of modified lipoproteins by scavenger receptors occurs mainly in macrophages, dendritic cells, and Kupffer cells of the liver. However, scavenger receptor expression also occurs in other cells, such as endothelial cells, aortic smooth muscle cells, neuronal cells, and keratinocytes. Thus, the local clearance of oxidized low‐density lipoprotein and the resolution of inflammatory processes may rely in part on the expression of scavenger receptors in “nonprofessional” phagocytes. Uptake of oxidized low‐density lipoprotein, without an efficient machinery to degrade them and uncontrolled expression of scavenger receptors, may lead to cellular deregulation, apoptosis, and formation of foam cells. Diseases accompanied by oxidation of lipoproteins, such as atherosclerosis, Alzheimer disease, glomerulosclerosis, ataxia with vitamin E deficiency, and possibly age‐dependent lipofuscin deposition, may share a common pathogenetic feature. This review will focus on foam cell formation, mainly within the atherosclerotic lesion, and the possible involvement of aberrant regulation of the scavenger receptor genes. To date, the regulatory mechanisms at the basis of scavenger receptor gene expression and their roles in atherosclerosis and other diseases are not well established. Knowledge on this subject could lead to a better understanding of the pathogenesis, prevention, and therapy of these diseases.
Genomic Organization of the Serine Protease Light Chain of Mouse Complement Factor I Gene
Authors: Jiang Ning , Molina Hector
Published: July 2002

A portion of the mouse complement factor I (mCFI) gene encoding for the mCFI light chain was cloned from a mouse 129/SVJ1 bacterial artificial chromosome library. It contains five exons and four introns. The intron sizes are remarkably different from the human homolog. Several polymorphisms were found in exon 13. One polymorphism was in the coding region, which causes a threonine in the Balb/c mCFI to be replaced by an isoleucine in the 129/SVJ1 mCFI. The other two polymorphisms are located in the 3′ untranslated region. The organization of the serine protease domain in mCFI is similar to that of trypsin but very different from that of the other complement serine proteases.
Enzymes Are Open Systems
Authors: Zahradník Frantisek Jan
Published: July 2002

The exchange of mass, energy, and information with the environment typical of open systems can also be found in enzymes. An enzyme is able to receive information on substrate concentration in an arbitrary concentration range. This actually follows from the elementary solution of Michaelis‐Menten kinetics. The Michaelis constant can be seen as a relation between the decreasing parameter of a zero‐order reaction and the increasing parameter of a first‐order reaction over a certain time interval. This excludes the state of zero‐order kinetics and consequently the state of zero content of concentration information.
The Nitrogen Source‐Dependent 126‐kDa Protein From Synechococcus PCC 7942 Plasmalemma: A Trimer of the NrtA Nitrate‐Binding Protein
Authors: Fresneau Chantal , Zinovieva Maria , Blanot Didier , Batelier GÉrard , Arrio Bernard
Published: July 2002

Expression of a 126‐kDa protein in the plasmalemma (cytoplasmic membrane) from Synechococcus PCC 7942 is dependent on the nitrogen source. Polyclonal antibody raised against the NrtA protein reacted with the 126‐kDa protein. Two peptide sequences from the 126‐kDa protein were retrieved in NrtA. FPLC purification of plasmalemma solubilised in Triton X‐100 gave a fraction consisting mainly of a 126‐kDa component (72.6%), as shown by sedimentation velocity. Equilibrium sedimentation of the same fraction gave evidence of the existence of an oligomeric structure (Ka = 2×10³ and 2.9 subunits). Thus, the 126‐kDa protein is considered as a trimeric arrangement of the 45‐kDa protein in the plasmalemma.
Targeting of an Intestinal Apical Endosomal Protein to Endosomes in Nonpolarized Cells
Authors: Wilson Jean M. , Colton Tamara L.
Published: July 2002

Polarized cells such as epithelial cells and neurons have distinct endosomal compartments associated with different plasma membrane domains. The endosomes of the neuronal cell body and the basolateral cytoplasm of epithelial cells are thought to perform cellular “housekeeping” functions such as the uptake of nutrients and metabolites, while the endosomes in the apical cytoplasm or axons are thought to be specialized for the sorting and transcytosis of cell type–specific ligands and receptors. However, it is not known if nonpolarized cells such as fibroblasts contain a specialized endosomal compartment analogous to the specialized endosomes found in neurons and epithelia. We have expressed a protein that is normally found in the apical early endosomes of developing intestinal epithelial cells in normal rat kidney fibroblasts. This apical endosomal marker, called endotubin, is targeted to early endosomes in transfected fibroblasts, and is present in peripheral as well as perinuclear endosomes. The peripheral endosomes that contain endotubin appear to exclude transferrin, fluid phase markers, and the mannose-6-phosphate receptor, although in the perinuclear region colocalization of endotubin and these markers is present. In addition, endotubin positive structures do not tubulate in response to brefeldin A and instead redistribute to a diffuse perinuclear location. Since this endosomal compartment has many of the characteristics of an apical or axonal endosomal compartment, our results indicate that nonpolarized cells also contain a specialized early endosomal compartment.
Characterization of the Adaptor-related Protein Complex, AP-3
Authors: Simpson Fiona , Peden Andrew A. , Christopoulou Lina , Robinson Margaret S.
Published: July 2002

We have recently shown that two proteins related to two of the adaptor subunits of clathrincoated vesicles, p47 (μ3) and β-NAP (β3B), are part of an adaptor-like complex not associated with clathrin (Simpson, F., N.A. Bright, M.A. West, L.S. Newman, R.B. Darnell, and M.S. Robinson, 1996. J. Cell Biol. 133:749–760). In the present study we have searched the EST database and have identified, cloned, and sequenced a ubiquitously expressed homologue of β-NAP, β3A, as well as homologues of the α/γ and σ adaptor subunits, δ and σ3, which are also ubiquitously expressed. Antibodies raised against recombinant δ and σ3 show that they are the other two subunits of the adaptor-like complex. We are calling this complex AP-3, a name that has also been used for the neuronalspecific phosphoprotein AP180, but we feel that it is a more appropriate designation for an adaptor-related heterotetramer. Immunofluorescence using anti-δ antibodies reveals that the AP-3 complex is associated with the Golgi region of the cell as well as with more peripheral structures. These peripheral structures show only limited colocalization with endosomal markers and may correspond to a postTGN biosynthetic compartment. The δ subunit is closely related to the protein product of the Drosophila garnet gene, which when mutated results in reduced pigmentation of the eyes and other tissues. Because pigment granules are believed to be similar to lysosomes, this suggests either that the AP-3 complex may be directly involved in trafficking to lysosomes or alternatively that it may be involved in another pathway, but that missorting in that pathway may indirectly lead to defects in pigment granules.
Conservation of the Centromere/Kinetochore Protein ZW10
Authors: Starr Daniel A. , Williams Byron C. , Li Zexiao , Etemad-Moghadam Bijan , Dawe R. Kelly , Goldberg Michael L.
Published: July 2002

Mutations in the essential Drosophila melanogaster gene zw10 disrupt chromosome segregation, producing chromosomes that lag at the metaphase plate during anaphase of mitosis and both meiotic divisions. Recent evidence suggests that the product of this gene, DmZW10, acts at the kinetochore as part of a tension-sensing checkpoint at anaphase onset. DmZW10 displays an intriguing cell cycle–dependent intracellular distribution, apparently moving from the centromere/kinetochore at prometaphase to kinetochore microtubules at metaphase, and back to the centromere/kinetochore at anaphase (Williams, B.C., M. Gatti, and M.L. Goldberg. 1996. J. Cell Biol. 134:1127-1140).

We have identified ZW10-related proteins from widely diverse species with divergent centromere structures, including several Drosophilids, Caenorhabditis elegans, Arabidopsis thaliana, Mus musculus, and humans. Antibodies against the human ZW10 protein display a cell cycle–dependent staining pattern in HeLa cells strikingly similar to that previously observed for DmZW10 in dividing Drosophila cells. Injections of C. elegans ZW10 antisense RNA phenocopies important aspects of the mutant phenotype in Drosophila: these include a strong decrease in brood size, suggesting defects in meiosis or germline mitosis, a high percentage of lethality among the embryos that are produced, and the appearance of chromatin bridges at anaphase. These results indicate that at least some aspects of the functional role of the ZW10 protein in ensuring proper chromosome segregation are conserved across large evolutionary distances.
Stu2p: A Microtubule-Binding Protein that Is an Essential Component of the Yeast Spindle Pole Body
Authors: Wang Peijing Jeremy , Huffaker Tim C.
Published: July 2002

Previously we isolated tub2-423, a cold-sensitive allele of the Saccharomyces cerevisiae gene encoding β-tubulin that confers a defect in mitotic spindle function. In an attempt to identify additional proteins that are important for spindle function, we screened for suppressors of the cold sensitivity of tub2-423 and obtained two alleles of a novel gene, STU2. STU2 is an essential gene and encodes a protein whose sequence is similar to proteins identified in a variety of organisms. Stu2p localizes primarily to the spindle pole body (SPB) and to a lesser extent along spindle microtubules. Localization to the SPB is not dependent on the presence of microtubules, indicating that Stu2p is an integral component of the SPB. Stu2p also binds microtubules in vitro. We have localized the microtubule-binding domain of Stu2p to a highly basic 100-amino acid region. This region contains two imperfect repeats; both repeats appear to contribute to microtubule binding to similar extents. These results suggest that Stu2p may play a role in the attachment, organization, and/or dynamics of microtubule ends at the SPB.
Processivity of the Motor Protein Kinesin Requires Two Heads
Authors: Hancock William O. , Howard Jonathon
Published: July 2002

A single kinesin molecule can move for hundreds of steps along a microtubule without dissociating. One hypothesis to account for this processive movement is that the binding of kinesin's two heads is coordinated so that at least one head is always bound to the microtubule. To test this hypothesis, the motility of a full-length single-headed kinesin heterodimer was examined in the in vitro microtubule gliding assay. As the surface density of single-headed kinesin was lowered, there was a steep fall both in the rate at which microtubules landed and moved over the surface, and in the distance that microtubules moved, indicating that individual single-headed kinesin motors are not processive and that some four to six single-headed kinesin molecules are necessary and sufficient to move a microtubule continuously. At high ATP concentration, individual single-headed kinesin molecules detached from microtubules very slowly (at a rate less than one per second), 100-fold slower than the detachment during two-headed motility. This slow detachment directly supports a coordinated, hand-over-hand model in which the rapid detachment of one head in the dimer is contingent on the binding of the second head.