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Neuronal activity related to eye-hand coordination in the primate premotor cortex.

To test the functional implications of gaze signals that we previously reported in the dorsal premotor cortex (PMd), we trained two rhesus monkeys to point to visual targets presented on a touch screen while controlling their gaze orientation. Each monkey had to perform four different tasks. To initiate a trial, the monkey had to put his hand on a starting position at the center of the touch screen and fixate a fixation point. In one task, the animal had to make a reaching movement to a peripheral target randomly presented at one of eight possible locations on a circle while maintaining fixation at the center of this virtual circle (central fixation + reaching). In the second task, the monkey maintained fixation at the location of the upcoming peripheral target and, later, reached to that location. After a delay, the target was turned on and the monkey made a reaching arm movement (target fixation + reaching). In the third task, the monkey made a saccade to the target without any arm movement (saccade). Finally, in the fourth task, the monkey first made a saccade to the target, then reached to it after a delay (saccade + reaching). This design allowed us to examine the contribution of the oculomotor context to arm-related neuronal activity in PMd. We analyzed the effects of the task type on neuronal activity and found that many cells showed a task effect during the signal (26/60; 43%), set (16/49; 33%) and/or movement (15/54; 28%) epochs, depending on the oculomotor history. These findings, together with previously published data, suggest that PMd codes limb-movement direction in a gaze-dependent manner and may, thus, play an important role in the brain mechanisms of eye-hand coordination during visually guided reaching.

Analysis of Variance↗

Detection of succinylacetone and the use of its measurement in mass screening for hereditary tyrosinemia.

A technique designed to measure quantitatively succinylacetone (4,6-dioxoheptanoic acid) is presented. It essentially involves the inhibition of delta-aminolevulinate dehydratase (EC 4.2.1.24) by succinylacetone. Prior to their use in the assay, the samples are heated at 100 degrees C for 30 min in order to transform all succinylacetoacetate (3,5-dioxooctanedioic acid) to succinylacetone. By this transformation of the first abnormal metabolite specific to hereditary tyrosinemia to the second and last one, which is a powerful inhibitor of delta-aminolevulinate dehydratase, we determine in one sensitive assay the total amount of both. Succinylacetone was measured in sera and urines from 19 patients with hereditary tyrosinemia. All sera and urines contained succinylacetone at concentrations ranging, respectively, from 2 to 100 mumol/l and from 190 to 6000 mumol/g creatinine. The technique was also adapted to dried blood spots on paper and was used as a test complementary to blood tyrosine determination in mass screening for hereditary tyrosinemia. A total of 2412 samples having concentrations of 60 mg/l or more of tyrosine were assayed, and ten showed the presence of succinylacetone. These were all from newborns with hereditary tyrosinemia. The test has proven to virtually eliminate false positives, and, thereby, much clerical work and parental anxiety.

Amino Acid Metabolism, Inborn Errors↗

Transcription factors do it together: the hows and whys of studying protein-protein interactions.

Protein-protein interactions are intrinsic to virtually every cellular process. Recent breakthroughs in techniques to study protein-interaction and the availability of fully sequenced plant genomes have attracted many plant scientists to undertake the first steps in the field of protein interactions. High-throughput screening systems allow the discovery of protein functions. Even without performing laborious functional assays, in planta functional homologues and redundant proteins can be accurately predicted based on protein-interaction maps. Therefore, protein-protein-interaction screenings are an essential supplement to the current functional-genomics toolbox.

DNA-Binding Proteins↗

Antimalarial and antitumor evaluation of novel C-10 non-acetal dimers of 10beta-(2-hydroxyethyl)deoxoartemisinin.

Four series of C-10 non-acetal dimers were prepared from key trioxane alcohol 10beta-(2-hydroxyethyl)deoxoartemisinin (9b). All of the dimers prepared displayed potent low nanomolar antimalarial activity versus the K1 and HB3 strains of Plasmodium falciparum. The most potent compound assayed was phosphate dimer 14a, which was greater than 50 times more potent than the parent drug artemisinin and about 15 times more potent than the clinically used acetal artemether. In contrast to their potent activity versus malaria parasites, virtually all of the dimers expressed poor anticancer activity apart from the trioxane phosphate ester dimers 14a and 14b, which expressed nanomolar growth inhibitory (GI50) values versus a range of cancer cell lines in the NCI 60 human cell line screen. Further detailed studies on these dimers in vitro in HL60 cells demonstrate that both phosphate ester dimers (14a and 14b) are more potent than the anticancer agent doxorubicin. Interestingly, phosphate ester monomers 9c and 9d, antimalarially active in the low nanomolar region versus P. falciparum, are inactive as anticancer agents even at concentrations in the millimolar region. This observation emphasizes the importance of two trioxane units for high antiproliferative activity, and we propose that the nature of the linker in dimers of this type plays a crucial role in imparting potent anticancer activity.

Acetals↗

[Magnetic resonance colonography: a new diagnostic tool].

Magnetic resonance colonography (MR colonography) is a three-dimensional imaging of the colon. Data postprocessing also allows for virtual colonoscopy, depicting the intraluminal morphology. Acquisition and postprocessing of high-resolution images require a state-of the-art modern MR scanner and a workstation. Regarding patient preparation, a bowel cleansing is necessary. The results of cancer screening have shown MR colonography to be superior to the traditional double-contrast barium enema. In inflammatory bowel disease, MR colonography can be used to assess disease activity, including spreading.

Colonic Neoplasms↗

[Human gingival fibroblast cultures for biological intra-oral material testing].

Cell cultures are useful screening methods for biocompatibility tests, as they provide information on the effects of test material on their metabolism. Primary cultures of human gingival fibroblasts play a special role, because they are organ-specific and because their metabolic state is virtually the same as that of in vivo cell material. Their characteristic growth pattern makes them excellent candidates for testing toxicity and stimulatory factors. Four methods were used for validating the data obtained. Possibilities of culturing these highly specialized cells are shown, techniques for assessing the cytotoxicity of test materials are presented and discussed.

Autoradiography↗

Endemic strongyloidiasis in northern Italy: clinical and immunological aspects.

One hundred and eighteen cases of S. stercoralis infection were detected by stool examination among 4203 patients seen over a period of three years at the Division of Infectious Diseases (IRCCS, San Matteo, University of Pavia) of a University Hospital in Northern Italy. Because chronic strongyloidiasis is often asymptomatic, a 3 per cent prevalence in this selected population indicates that the parasitosis may be much more frequent in the general population of the area. Parasite-specific humoral responses were studied in 48 patients. Strongyloides-specific IgG and IgA antibodies were found by ELISA in the serum of 40 (83.3 per cent) and 43 (89.5 per cent) patients, respectively, and specific IgE antibodies were detected by RAST in 39 (81.3 per cent). The specific IgG antigen recognition patterns of these patients were compared by immunoblotting with those of North and South American and Thai patients, and differences among the groups suggested the existence of separate geographic strains of S. stercoralis. This was further supported by the virtual absence in our patients of the larva currens, a common and highly characteristic sign of strongyloidiasis in subjects infected in southeast Asia. More awareness of the epidemiologic and clinical features of strongyloidiasis, with the use of serologic tests to screen candidates for immunosuppression, may result in the early detection of chronic uncomplicated strongyloidiasis and in the prevention of fatal opportunistic hyperinfections.

Adult↗

Identification of the second common Jewish Gaucher disease mutation makes possible population-based screening for the heterozygous state.

Gaucher disease is an autosomal recessive glycolipid storage disease characterized by a deficiency of glucocerebrosidase. The disease is most common in persons of Ashkenazi Jewish ancestry and the most common mutation, accounting for about 75% of the mutant alleles in this population, is known to be an A----G substitution at cDNA nucleotide (nt) 1226. Screening for this disease has not been possible because nearly 25% of the mutant alleles had not been identified, but linkage analysis led to the suggestion that most of these could be accounted for by a single mutation. We now report the discovery of this mutation. The insertion of a single nucleotide, a second guanine at cDNA nt 84 (the 84GG mutation), has been detected in the 5' coding region of the glucocerebrosidase gene. The amount of mRNA produced is shown to be normal but since the frameshift produced early termination, no translation product is seen. This finding is consistent with the virtual absence of antigen found in patients carrying this mutation. The 84GG mutation accounts for most of the previously unidentified Gaucher disease mutations in Jewish patients. The common Jewish mutation at nt 1226, the 84GG mutation, and the less-common mutation at nt 1448 accounted for 95% of all of the Gaucher disease-producing alleles in 71 Jewish patients. This now makes it possible to screen for heterozygotes on a DNA level with a relatively low risk of missing couples at risk for producing infants with Gaucher disease.

Age Factors↗

(TG)n uncovers a sex-specific hybridization pattern in cattle.

Screening of a bovine genomic library with the human minisatellite 33.6 probe uncovered a family of clones that, when used to probe Southern blots of bovine genomic DNA digested with the restriction enzyme HaeIII or MboI, revealed sexually dimorphic, but otherwise virtually monomorphic, patterns among the larger DNA fragments to which they hybridized. Characterization of one of these clones revealed that it contains different minisatellite sequences. The sexual dimorphism hybridization pattern observed with this clone was found to be due to multiple copies of two tandemly interspersed repeats: the simple sequence (TG)n and a previously undescribed 29-bp sequence. Both repeats appear to share many genomic loci including autosomal loci. In contrast, Southern analysis of AluI- or HinfI-digested bovine DNA with the (TG)n repeat used as a probe yielded substantial polymorphism. These results show that (i) different minisatellites can be found in a cluster, (ii) both simple and more complex repeated sequences other than the simple quaternary (GATA)n repeat can be sexually dimorphic, and (iii) simple repeats can reveal substantial polymorphism.

Animals↗

Stability of sex differences in cognition in advanced old age: the role of education and attrition.

We examined whether patterns of sex differences on tasks of perceptual speed, episodic memory, verbal fluency, and verbal knowledge are maintained during advanced old age. Using incomplete 13-year longitudinal data from participants in the Berlin Aging Study screened for dementia (N = 368; M = 83 years; range 70-100 years at baseline assessment), we estimated sex-specific age trajectories of cognitive change and explored the contributing role of education and attrition. We found that women and men declined virtually in parallel, with no evidence of differential change. After we controlled for age cohort-related differences in education, women outperformed men on tasks in the four cognitive domains. Findings also provide initial evidence that sex differences might be masked by differential patterns of sample attrition.

Aged↗

Targeting performance of three large-scale, nutrition-oriented social programs in Central America and Mexico.

This study evaluated whether three nutrition-oriented programs in Central America and Mexico have been successful in targeting those households most vulnerable to undernutrition and poverty. For each country, nationally representative data sets were used to estimate cutoff points dividing the population into 10 equal-sized groups according to child anthropometric measurements (age-standardized height) and household income (per capita household expenditures). Households meeting eligibility criteria were then assessed using special baseline surveys or national data obtained before implementation of the program. Children in these households were classified according to national deciles of height-for-age, and households were classified according to expenditure deciles. In spite of markedly differing targeting strategies, each of the programs was well targeted, with 45% (Honduras and Mexico) and 52% (Nicaragua) of children in eligible households coming from the lowest two deciles of the national distributions, and virtually none from the upper two deciles. Similar results pertained to household income. These experiences demonstrate that vulnerable households can be targeted relatively straightforwardly, and that the need to do this does not in every case imply household-level income screening.

Adult↗

PCR amplification, cloning, and construction of HIV-1 infectious molecular clones from virtually full-length HIV-1 genomes.

The development of mixtures of highly processive and high-fidelity thermostable DNA polymerases has enabled the routine recovery of DNA sequences in excess of 25 kb generated by polymerase chain reaction. This powerful tool has been instrumental in the ability to recover virtually full-length HIV-1 proviral DNA as a single, contiguous fragment. Such fragments allow for the clean interpretation of the genomic organization of HIV-1 provirus, as they are not confounded by molecular mosaicism that accrues to overlapping subgenomic amplification strategies. We detail here a robust procedure to produce virtually full-length, single contiguous 9.2-kb HIV-1 amplimers whose full-length infectious potential is reconstituted upon cloning into long terminal repeat-replacement vectors. Large numbers of HIV-1 proviral clones can now be quickly generated and screened to identify the fraction of the viral quasispecies with the highest capacity for viral replication. The methods used to construct long terminal repeat-replacement vectors, amplify HIV-1 provirus, reconstitute full-length provirus, and recover viral stocks will be illustrated using a circulating recombinant form 1 (CRF01_AE, formerly known as subtype E) primary isolate.

Cloning, Molecular↗

[Facial telethermography in the diagnosis of carotid insufficiency in the extracranial sector].

The usefulness of facial telethermography in the diagnosis of circulatory insufficiency owing to occlusion of the extracranial vessels is discussed. The method has proved of great assistance in the screening of patients with vascular diseases, and in the choice of a subsequent contrastographic examination. It gave a significant number of positive findings in cases of insufficiency of the internal carotid artery. The number of false positives and false negatives was virtually negligible.

Arterial Occlusive Diseases↗

Hyperhomocysteinaemia: a newly recognized risk factor for vascular disease.

In the past decade significant progress has been made in understanding of hyperhomocysteinaemia and its association with the proneness to premature development of vascular disease. Pooled data from a large number of studies demonstrate that mild hyperhomocysteinaemia after standardized methionine loading is present in 21% of patients with coronary artery disease, in 24% of patients with cerebrovascular disease, and in 32% of patients with peripheral vascular disease. A relative risk of 13.0 (95% confidence interval 5.9-28.1) of vascular disease at relatively young age can be calculated in subjects with such abnormal response to methionine loading. Pathological homocysteine levels are affected by genetic defects in homocysteine metabolism which have still not been completely clarified and which are more complex than originally supposed. Furthermore, a variety of non-genetic determinants such as deficiency of folate or vitamin B12 has to be taken into account. Mild hyperhomocysteinaemia can be reduced to normal in virtually all cases by simple and safe treatment with vitamin B6, folic acid, and betaine, each of which is involved in methionine metabolism. A clinically beneficial effect of such an intervention, which is currently under investigation, could make large-scale screening mandatory for this risk factor.

Arteriosclerosis↗

A comparison of meconium, maternal urine and neonatal urine for detection of maternal drug use during pregnancy.

A large scale drug screening study was done to determine the prevalence of drug use in a large metropolitan, obstetric population. Meconium and first voided urine, as well as maternal urine were collected from 423 consecutive deliveries. Urine samples and methanolic extracts of meconium were initially screened by Enzyme Multiplied Immunoassay Technique (EMIT) and then confirmed by Gas Chromatography/Mass Spectrometry (GC/MS). Analysis of cocaine metabolite as benzoylecogonine, cannabinoid as carboxy-THC, codeine, morphine and methadone were included in the study. The positive rate for benzoylecgonine was virtually identical for meconium, maternal urine and neonatal urine (12%). Analysis of meconium was found to be more reliable than analysis of maternal or neonatal urine for the detection of benzoylecgonine. Meconium did not appear to offer an advantage over maternal or neonatal urine for detection of cannabinoid, codeine, morphine, or methadone.

Adult↗

Stroke incidence, prevalence, and survival: secular trends in Rochester, Minnesota, through 1989.

BACKGROUND AND PURPOSE: Trends in stroke incidence and mortality are available from few sites worldwide. The unique nature of the medical care in patients from Rochester, Minn, has allowed the ascertainment of virtually all cases of stroke for community residents for the period dating from 1955. This study updates incidence rates, stroke mortality, radiological imaging availability, and prevalence for stroke and subtypes of stroke though 1989 and evaluates trends since 1955. METHODS: Medical records of all residents of Rochester with potential diagnosis of stroke during the 5-year period through December 31, 1989 were screened to determine whether the case met the criteria for stroke. The type of stroke was determined with the use of all available data. Average annual age- and sex-adjusted incidence rates for stroke and various types of stroke were calculated for 5-year periods from 1955 to 1989. Prevalence of stroke was also determined. Poisson regression was used to better clarify the dependence of stroke incidence on multiple variables. Cox proportional hazards modeling evaluated trends in short-term and long-term stroke survival. RESULTS: We found that 496 incidence cases of stroke occurred among Rochester residents during 1985 to 1989. The annual age- and sex-adjusted stroke incidence rates was 145 per 100 000 population, which was virtually unchanged from 1980 to 1984 and 13% higher than the rate determined in 1975 to 1979. Increasing incidence rates compared with the 1970s were noted in all groups aged older than 54 years and in both sexes. Survival after cerebral infarction was dependent on age and calendar year when trends dating from 1955 were evaluated. The annual incidence rate of intraparenchymal hemorrhage was twice that of subarachnoid hemorrhage. CONCLUSIONS: Incidence rates for stroke have remained at a high rate than those determined in the 1970s. Although the occurrence of untreated hypertension has been reported to be stable or decreasing, the incidence rate of stroke is higher than those noted during the decline in stroke incidence during the 1960s and 1970s. In addition to the impact of radiological imaging, increased contribution of patients with ischemic heart disease, and the possible detection of milder cases of stroke, other factors, which are undefined, may be contributing to the increased stroke incidence rates detected over the last 10 years.

Adult↗

Direct imaging of explosives.

Any technique that can detect nitrogen concentrations can screen for concealed explosives. However, such a technique would have to be insensitive to metal, both encasing and incidental. If images of the nitrogen concentrations could be captured, then, since form follows function, a robust screening technology could be developed. However these images would have to be sensitive to the surface densities at or below that of the nitrogen contained in buried anti-personnel mines or of the SEMTEX that brought down Pan Am 103, approximately 200 g. Although the ability to image in three-dimensions would somewhat reduce false positives, capturing collateral images of carbon and oxygen would virtually assure that nitrogenous non-explosive material like fertilizer, Melmac dinnerware, and salami could be eliminated. We are developing such an instrument, the Nitrogen Camera, which has met experimentally these criteria with the exception of providing oxygen images, which awaits the availability of a sufficiently energetic light source. Our Nitrogen Camera technique uses an electron accelerator to produce photonuclear reactions whose unique decays it registers. Clearly if our Nitrogen Camera is made mobile, it could be effective in detecting buried mines, either in an active battlefield situation or in the clearing of abandoned military munitions. Combat operations require that a swathe the width of an armored vehicle, 5 miles deep, be screened in an hour, which is within our camera's scanning speed. Detecting abandoned munitions is technically easier as it is free from the onerous speed requirement. We describe here our Nitrogen Camera and show its 180 pixel intensity images of elemental nitrogen in a 200 g mine simulant and in a 125 g stick of SEMTEX. We also report on our progress in creating a lorry transportable 70 MeV electron racetrack microtron, the principal enabling technology that will allow our Nitrogen Camera to be deployed in the field.

Explosions↗

Preparticipation orthopedic screening evaluation.

OBJECTIVE: To review evidence-based support for the preparticipation orthopedic evaluation. DATA SOURCES/METHODS: Articles were reviewed that dealt with the sensitivity, specificity, and predictive value of the components of the standardized preparticipation orthopedic evaluation. In addition, studies describing musculoskeletal conditions/findings predictive of future injuries were sought through a PubMed search. RESULTS: The sensitivity of the evaluation questionnaire appears to be adequate, exceeding 90% in some studies. There is little or no published information documenting that the physical examination (1) approaches the questionnaire in either sensitivity or specificity or (2) identifies elements of value based on their association with future injuries or reinjuries. There are no readily discernible elements even in an expanded examination that are documented as being predictive of future problems. CONCLUSIONS: The current questionnaire and examination appear to fulfill adequately both legal and institutional requirements. Practitioners should be aware of the absence of data linking virtually any of the findings on the examination to either an increase or a decrease in the likelihood of future injuries. There is no evidence that increasing the scope of the examination would enhance its predictive value.

Evidence-Based Medicine↗