Influence of additives to the formulation of n.c.a. [¹¹C]PiB on sterile filter performance.

The influence of different additives (PEG 300, PEG 400, PG) to the product solution of [(11)C]PiB was investigated with regard to tracer retention for a number of commonly used sterile filters for aseptic manufacturing of PET-tracers. The effect of the amount of additive with regard to tracer retention and the resulting viscosity of the filtration solution was determined. Recommendations for the individual combinations of filters and amounts of additives suitable for the different filtration methods that are implemented in commercially available synthesis modules are given as well.

Intensity of parasitic mite infection decreases with hibernation duration of the host snail.

Temperature can be a limiting factor on parasite development. Riccardoella limacum, a haematophagous mite, lives in the mantle cavity of helicid land snails. The prevalence of infection by R. limacum in populations of the land snail Arianta arbustorum is highly variable (0-78%) in Switzerland. However, parasitic mites do not occur in host populations at altitudes of 1290 m or higher. It has been hypothesized that the host's hibernation period might be too long at high elevations for mites and their eggs to survive. To test this hypothesis, we experimentally infected snails and allowed them to hibernate at 4°C for periods of 4-7 months. Winter survival of host snails was negatively affected by R. limacum. The intensity of mite infection decreased with increasing hibernation duration. Another experiment with shorter recording intervals revealed that mites do not leave the host when it buries in the soil at the beginning of hibernation. The number of mites decreased after 24 days of hibernation, whereas the number of eggs attached to the lung tissue remained constant throughout hibernation. Thus, R. limacum survives the winter in the egg stage in the host. Low temperature at high altitudes may limit the occurrence of R. limacum.

Efficacy and safety of sirolimus and everolimus in heart transplant patients: a retrospective analysis.

BACKGROUND: Since its introduction as an immunosuppressant in the late 1990s, sirolimus (SRL) has been used to prevent rejections after heart transplantation (HTx) in the United States. An analogue, everolimus (ERL) has been mainly used in Europe. We performed a retrospective longitudinal single-center study to evaluate efficacy and side effects of SRL and ERL. PATIENTS AND METHODS: We analyzed 71 patients, 39 in the SRL and 32 in the ERL group. The following data were collected: Trough levels of SRL and ERL, biopsy-proven rejections, renal function, blood lipids, hematology, blood pressure, pulse rate, and side effects (via an anonymous questionnaire). Follow-up time was 6 months. No prisoners or organs from prisoners were used in the study. RESULTS: Introduction of SRL or ERL into therapy took place 44 or 42 months (average) after HTx. SRL and ERL were equally effective in preventing rejection (8/39 versus 6/32). Hemoglobin levels decreased slightly in the SRL group (nonsignificant). Leucocytes and thrombocyte levels decreased in both groups (P < .05 only in the ERL group). Creatinine levels remained unchanged. Cholesterol and triglyceride levels increased significantly in the SRL group. High-density lipoprotein levels increased significantly in the ERL group. Vital signs remained stable in both groups. Side effects (mainly edema, gastrointestinal symptoms and infections) were considerable and prompted discontinuation in 39% of all patients in both groups. Infections were more frequent in SRL (18/39 versus 12/32, nonsignificant). Calcineurin therapy could be reduced by 25% in SRL and 45% in ERL. CONCLUSION: The impact of SRL and ERL on laboratory values and rejection rates, as well as on clinical parameters, is similar with a slight advantage to ERL regarding lipids and rate of infections (not significant). Both SRL and ERL allow an important reduction of calcineurin-therapy; however, both drugs have considerable side effects, which often require discontinuation of therapy.

Writing kinematics and pen forces in writer's cramp: effects of task and clinical subtype.

OBJECTIVE: Writer's Cramp (WC) is defined as a task-specific form of focal-hand-dystonia generating hypertonic muscle co-contractions resulting in impaired handwriting. Little is known about kinematic and dynamic characteristics in handwriting in the different subtypes of WC. METHODS: In this study, kinematic and force analyses were used to compare handwriting capacity of 14 simple, 13 dystonic WC-patients and 14 healthy subjects. The effect of task-complexity was investigated using a simple repetitive writing-task, writing pairs of letters, a sentence and copying a text. RESULTS: In general, patients showed significant deficits in kinematic and force parameters during writing, but no consistent differences between the two subtypes of WC were found. The complexity of writing material modulated writing parameters in all groups, but less complex material did not ameliorate the patients' deficits relative to control subjects. CONCLUSIONS: The similarity of deficits in patients with simple and dystonic WC does not support the concept of a unitary progression of deficits causing a switch from simple to dystonic WC. Dystonic WC seems to be characterized by a spread of symptoms independent of severity. Obviously, the deficits concern elementary aspects of writing and are not modulated by more complex aspects. SIGNIFICANCE: Quantification of writing deficits by simple and short phrases with kinematic and force parameters can substantially improve the characterization of WC.

Contact-based transmission models in terrestrial gastropod populations infected with parasitic mites.

Parasite transmission fundamentally affects the epidemiology of host-parasite systems, and is considered to be a key element in the epidemiological modelling of infectious diseases. Recent research has stressed the importance of detailed disease-specific variables involved in the transmission process. Riccardoella limacum is a hematophagous mite living in the mantle cavity of terrestrial gastropods. In this study, we experimentally examined whether the transmission success of R. limacum is affected by the contact frequency, parasite load and/or behaviour of the land snail Arianta arbustorum, a common host of R. limacum. In the experiment the transmission success was mainly affected by physical contacts among snails and slightly influenced by parasite intensity of the infected snail. Using these results we developed two different transmission models based on contact frequencies and transmission probability among host snails. As parameters for the models we used life-history data from three natural A. arbustorum populations with different population densities. Data on contact frequencies of video-recorded snail groups were used to fit the density response of the contact function, assuming either a linear relationship (model 1) or a second-degree polynomial relationship based on the ideal gas model of animal encounter (model 2). We calculated transmission coefficients (beta), basic reproductive ratios (R(0)) and host threshold population densities for parasite persistence in the three A. arbustorum populations. We found higher transmission coefficients (beta) and larger R(0)-values in model 1 than in model 2. Furthermore, the host population with the highest density showed larger R(0)-values (16.47-22.59) compared to populations with intermediate (2.71-7.45) or low population density (0.75-4.10). Host threshold population density for parasite persistence ranged from 0.35 to 2.72 snails per m(2). Our results show that the integration of the disease-relevant biology of the organisms concerned may improve models of host-parasite dynamics.

Novel microsatellite loci in the grass snake (Natrix natrix) and cross-amplification in the dice snake (Natrix tessellata).

Six novel polymorphic microsatellite loci are presented for the grass snake (Natrix natrix), a species with declining populations in many regions. The number of alleles per locus ranged from two to seven. Four dice snake (Natrix tessellata) microsatellites were polymorphic in the grass snake with three to four alleles. At two loci, the expected heterozygosity differed significantly from observed heterozygosity. Cross-amplification of the grass snake markers in the dice snake showed two polymorphic microsatellites with two and four alleles.

Experimental evidence for a new transmission route in a parasitic mite and its mucus-dependent orientation towards the host snail.

The route of transmission and host finding behaviour are fundamental components of a parasite's fitness. Riccardoella limacum, a haematophagous mite, lives in the mantle cavity of helicid land snails. To date it has been assumed that this parasitic mite is transmitted during courtship and mating of the host. Here we present experimental evidence for a new transmission route in the host snail Arianta arbustorum. Parasite-free snails were kept on soil on which previously infected host snails had been maintained for 6 weeks. R. limacum was successfully transmitted via soil without physical contact among hosts in 10 out of 22 (45.5%) cases. In a series of experiments we also examined the off-host locomotion of R. limacum on snail mucus and control substrates using an automated camera system. Parasitic mites showed a preference to move on fresh mucus. Our results support the hypothesis that R. limacum uses mucus trails to locate new hosts. These findings should be considered in commercial snail farming and when examining the epidemiology of wild populations.

Syndromic approach for determination of reproductive tract infections among adolescent girls.

The present study shows overall prevalence (64%) of reproductive tract infection among adolescent girls, based on self-perceived symptoms. Mean age of respondents were found to be 17.8 +/- 0.82 years and mean age at marriage and mean age at first pregnancy were 17.2 years and 17.5 years respectively; 35.35% of girls in the present study were married. In addition, no significant difference was observed between unmarried (60.10%) and married (71.17%) reproductive tract infection groups. Moreover, no significant association was present in prevalence of reproductive tract infection between the Muslim (67%) and the Hindu (60%). Highest prevalence (84.06%) of reproductive tract infection was observed among illiterate girls and with improvement of educational status there was decrease in the prevalence and the association was found highly significant. Significantly, higher prevalence (72%) was observed among members of family size 7 and above.

Coevolution of male and female reproductive traits in a simultaneously hermaphroditic land snail.

Inter- and intraspecific studies in gonochoristic animals reveal a covariation between sperm characteristics and the size of the female reproductive tract, indicating a rapid evolutionary divergence, which is consistent with the theory of post-copulatory sexual selection. Simultaneous hermaphrodites differ from species with separate sexes (gonochorists) in that they possess both functional male and female reproductive organs at the same time. We investigated whether in hermaphroditic animals intraspecific variation in reproductive traits results from divergent coevolution, by quantifying the variation in male and female traits among six natural populations of the snail Arianta arbustorum and examining the covariation in interacting traits. There was a significant among-population variation in spermatophore volume, number of sperm transferred and sperm length, as well as in volume of the sperm storage organ (spermatheca) and number of tubules, but not in spermatheca length. We found a positive association between sperm number transferred and spermatheca volume. This result suggests that the same post-copulatory mechanisms as in gonochorists drive the correlated evolution of reproductive characters in hermaphrodites.

An epidemiological study on risk factors of diabetes mellitus among patients attending a medical college hospital in Kolkata, West Bengal.

Two hundred and thirty cases of diabetes mellitus were selected in a case-control fashion by applying systemic random sampling technique taking 20% of the patients attending the diabetic clinic of Calcutta National Medical College during the period from January to June, 2003 to find out association and risks of some socio-economic and demographic factors with the occurrence of diabetes mellitus. Controls were selected from the patients admitted in the department of surgery in the same period. Factors like physical activity, socio-economic status, residential status, family history and obesity have shown statistically significant association with diabetes. For physical activity the estimated relative risk is maximum in case of sedentary worker where the odd's ratio was 9.4. So far other factors are concerned the estimated relative risk of diabetes was 5.1 in case of urban population, 6.2 in case of positive family history, 2.5 in case of upper and upper middle classes, and 2.4 in case of obesity in comparison to their respective reference category.

Mitochondrial differentiation in a polymorphic land snail: evidence for Pleistocene survival within the boundaries of permafrost.

The genetic differentiation of populations having colonized formerly unsuitable habitats after the Pleistocene glaciations depends to a great extent on the speed of expansion. Slow dispersers maintain their refugial diversity whereas fast dispersal leads to a reduction of diversity in the newly colonized areas. During the Pleistocene, almost the entire current range of the land snail Arianta arbustorum has repeatedly been covered with ice or been subjected to permafrost. Owing to the low potential for dispersal of land snails, slow (re)colonization of the wide range from southern refugia can be excluded. Alternatively, fast, passive dispersal from southern refugia or survival in and expansion from multiple refugia within the area subjected to permafrost may account for the current distribution. To distinguish between these scenarios we reconstructed a phylogeography based on the sequences of a fragment of the cytochrome oxidase I from 133 individuals collected at 45 localities and analysed the molecular variance. Seventy-five haplotypes were found that diverged on average at 7.52% of positions. This high degree of diversity suggests that A. arbustorum is an old species in which the population structure, isolation and the hermaphroditic nature have reduced the probability of lineage extinction. The genetic structure was highly significant with the highest variance partition found among regions. Geographic distance and mitochondrial differentiation were not congruent. Lineages had overlapping ranges. The clear genetic differentiation and the patchy pattern of haplotype distribution suggest that colonization of formerly unsuitable habitats was mainly achieved from multiple populations from within the permafrost area.

Neurological and neuropsychological examination and outcome after use of an intra-aortic filter device during cardiac surgery.

Cerebral embolization of particles after cardiac surgery is frequently associated with neurological deficits. Aortic crossclamp manipulation seems to be the most significant cause of emboli release during cardiac surgery. The goal of this study was to demonstrate whether the use of an intra-aortic filter device has an effect on the magnet resonance imaging (MRI) and functional neurological outcome. Twenty-four patients undergoing cardiosurgical procedures using cardiopulmonary bypass (CPB) were selected: coronary artery bypass graft (CABG) surgery (n = 17), aortic valve replacement (AVR) surgery (n = 4) or combined procedures (n = 3). Patients were evaluated by diffusion weighted MRI of the brain, neurological examination and neuropsychological assessment regarding alertness as well as divided and selective attention before and five to seven days after surgery. The patients were divided into two groups. In group I, 12 patients received a filter through a modified 24 F arterial cannula immediately before the aortic crossclamp was released. Filters remained in the aorta until CPB was discontinued. Intraoperatively, bilateral middle cerebral artery transcranial Doppler (TCD) was monitored at baseline, at the beginning of CPB, at a timepoint when the aorta was crossclamped, when the filter was inserted and while the crossclamp was switched to partial clamping until the CPB was discontinued. TCD was used for detection of microembolic signals (MES). The captured material in the filter was examined histologically. Twelve patients served as controls without aortic filtration (group II). The MRI of the brain did not show any diffusion alterations in either group before or after surgery. No patient developed a focal neurological deficit or stroke. Intraoperative quantitative MES detection revealed a four to tenfold increase in patients of group I compared with group II (5-6 versus 0.5-1 MES/min) during the filter dwell time. There was no consistent pattern regarding the neurobehavioural sequelae. Filters showed arteriosclerotic debris in 75% of the patients. The use of the intra-aortic filter device did not show a positive effect on neurological, neuroradiographical and neuropsychological outcomes. The increase of the MES rate in group I patients may be due to microbubbles generated as microcavitations by the filter or the aortic filter cannula. The intra-aortic filter was able to capture atheromatous material in 75% of the patients.

A study on awareness of AIDS among school students and teachers of higher secondary schools in north Calcutta.

Higher Secondary School students and their teachers were studied to assess the knowledge about AIDS and attitude towards AIDS patients. Only 13.5% senior school students and 16.2% teachers had clear knowledge regarding AIDS--its general aspects, transmission and prevention. Girls had higher and clear knowledge than boys. 45.8% of girls, 38.8% of boys students and 20.3% of teachers had positive attitudes towards nursing an AIDS case. It is suggested that schools have to device ways to open up more effective communication with students in relation to education on sex and AIDS. Training on AIDS should be emphasized on school teachers who on their turn can teach the students in a correct way about AIDS.

Coverage of Maternal Care Services in the state of West Bengal.

Evaluation of the Coverage of Maternal Care Services was conducted during the year 1999 in the entire State of West Bengal, Urban poor localities of randomly selected three Municipal Corporation areas and 'high-risk' urban wards or villages of West Bengal that had reported AFP cases in 1998. It was observed that mothers who had three or more antenatal check-ups varied between 54% to 82% in different study areas. Tetanus Toxoid coverage varied between 83.5% to 93.4% being lowest in high-risk areas. Only a very small section of mothers (12.7% to 23.7%) consumed 100 or more Iron and Folic acid tablets during pregnancy. 'Danger Signs' of pregnancy were informed to only 21.1% to 38.2% of the pregnant mothers. Proportion of home deliveries varied between 16.7% (Calcutta) to as high as 72.7% (high-risk areas). Deliveries conducted by untrained personnel were 60.8% in high risk areas, 38.1% in State clusters and between 14.2% to 29.4% in the three urban areas. Vast majority of mothers (78.8% to 88.4%) received no post-natal check-up.

Plastidic metabolite transporters and their physiological functions in the inducible crassulacean acid metabolism plant Mesembryanthemum crystallinum.

The inducible crassulacean acid metabolism (CAM) plant Mesembryanthemum crystallinum accumulates malic acid during the night and converts it to starch during the day via a pathway that, because it is located in different subcellular compartments, depends on specific metabolite transport across membranes. The chloroplast glucose transporter (pGlcT) and three members of the phosphate translocator (PT) family were isolated. After induction of CAM, transcript amounts of the phosphoenolpyruvate (PEP) phosphate translocator (PPT) and the glucose-6-phosphate (Glc6P) phosphate translocator (GPT) genes were increased drastically, while triose phosphate (TP) phosphate translocator (TPT) and the pGlcT transcripts remained unchanged. PPT- and GPT-specific transcripts and transporter activities exhibited a pronounced diurnal variation, displaying the highest amplitude in the light. pGlcT transcripts were elevated towards the end of the light period and at the beginning of the dark period. These findings, combined with diurnal variations of enzyme activities and metabolite contents, helped to elucidate the roles of the PPT, GPT, TPT and pGlcT in CAM. The main function of the PPT is the daytime export from the stroma of PEP generated by pyruvate orthophosphate:dikinase (PPDK). The increased transport activity of GPT in the light suggests a higher requirement for Glc6P import for starch synthesis rather than starch mobilization. Most likely, Glc6P rather than 3-phosphoglycerate or triose phosphates is the main substrate for daytime starch biosynthesis in M. crystallinum plants in which CAM has been induced (CAM-induced), similar to non-green plastids. In the dark, starch is mobilized both phosphorylytically and amylolytically and the products are exported by the GPT, TPT and pGlcT. The transport activities of all three phosphate translocators and the transcript amounts of the pGlcT adapt to changing transport requirements in order to maintain high metabolic fluxes during the diurnal CAM cycle.

A novel type of mutation in the cysteine rich domain of the RET receptor causes ligand independent activation.

Multiple endocrine neoplasia type 2A (MEN 2A) is a dominantly inherited cancer syndrome, which involves the triad of MTC, pheochromocytoma, and hyperparathyroidism. Missense mutations in one of six cysteine codons in the extracellular cysteine-rich domain of the RET proto-oncogene predispose to this disease. These mutations cause ligand-independent constitutive activation of the tyrosine kinase receptor by the formation of disulfide-bonded homodimers. We examined a different type of mutation, which results in an additional cysteine in the cysteine rich domain. A duplication of 9 bp in the first case resulted in an insertion of three amino acids between codon 633 and 634. In the second case a 12 bp duplication in exon 11 results in four additional amino acids between codon 634 and 635. Here we demonstrate that an additional cysteine causes a ligand independent dimerization of the RET receptor in transfected NIH3T3 cells, which results in an activation of the intracellular tyrosine kinase.

Biotin and biocytin uptake into cultured primary calf brain microvessel endothelial cells of the blood-brain barrier.

The uptake of biotin and the closely related biocytin was characterized in primary cultures of calf brain microvessel endothelial (CBME) cells. Biotin uptake was found to be Na(+)-gradient dependent and independent of changes in the membrane potential. Concentration dependence revealed a single saturation mechanism with a K(m) of 47 microM and a V(max) of 101 pmol/min/mg. Inhibition studies demonstrated dependence on metabolic energy and the necessity for a free carboxyl group for transport activity. The anticonvulsants primidone and carbamazepine had no inhibitory effect. Biotin uptake into CBME cells is a secondary active, electroneutral, saturable and specific process. Biocytin which accumulates in biotinidase deficiency, a human congenital disorder, did not inhibit biotin uptake and was not transported into these cells. The presence of human serum with normal biotinidase activity significantly reduced biotin uptake by about 50%. Further, added biocytin was hydrolyzed to biotin, which accumulated intracellularly but to a lesser extent than added free biotin. Biotin uptake after addition of plasma of biotinidase-deficient patients was not different from that in the presence of normal serum. These results indicate that the absence of biotinidase activity in serum does not reduce blood-brain barrier transport of biotin.

Consumption rates and food preferences of slugs in a calcareous grassland under current and future CO2 conditions.

This study explored consumption of a generalist herbivore feeding on leaf tissue of various plant species of a calcareous grassland, and tested whether consumption levels and preferences changed when plants were exposed to 5 years of in situ CO2 enrichment. The first part of this experiment tested whether the consumption patterns of slugs (Deroceras reticulatum) observed in single-species feeding tests were altered when slugs were given a choice of food sources. Overall consumption increased 270% when slugs were given a choice, and they preferred having a choice of food sources more than they preferred having any one food source. Surprisingly, slugs consumed fewer legumes and grasses and more non-leguminous forbs when given a choice. In the second part of this experiment, feeding behaviors of slugs in response to elevated CO2 were investigated by feeding them leaves of two legumes, one grass, and a non-leguminous forb (Trifolium medium, Lotus corniculatus, Bromus erectus, and Sanguisorba minor, respectively) in two or four species combinations. In the leguminous species mix, the non-leguminous species mix, and the combined mix (legumes and non-legumes), neither overall consumption by herbivores nor species preference was significantly altered by long-term CO2 enrichment. In the combined species mix, slugs preferred legumes to non-legumes (P=0.012) and exhibited a weak functional group preference shift from non-legumes to legumes (P=0.089) in response to CO2 enrichment. This is the first time such a shift has been observed, and provides evidence that there may be multiple herbivore responses to rising atmospheric CO2 concentrations. Numerous single-species feeding tests using insects have shown that consumption by herbivores may increase when herbivores are fed plants grown in enriched CO2 atmospheres. This study clearly demonstrates the limited applicability of non-choice feeding trials to generalist herbivores in species-rich communities.

Sperm allocation in the simultaneously hermaphroditic land snail Arianta arbustorum.

To test the idea that individuals of the simultaneously hermaphroditic land snail Arianta arbustorum can control the number of spermatozoa in their spermatophores, we investigated whether they differentially release sperm to virgin or nonvirgin partners with respect to the potential risk of sperm competition in a given mating. The number of sperm transferred ranged from 802 620 to 3 968 800 (X= 2 185 100; N=91), but was related neither to the mating history of the partner nor to copulation duration. This indicates that individuals of A. arbustorum are not able to adjust sperm expenditure to the mating history of the partner. Furthermore, the number of sperm transferred was correlated neither with the size of the donor nor with the size of the recipient. It has been proposed that the sexual conflict between the two genders in simultaneous hermaphrodites could be resolved by gamete trading. Theory predicts that sperm trading should occur in hermaphrodites in which the female role controls fertilization, for example in gastropods with a gametolytic gland and/or sperm storage such as A. arbustorum. To see whether sperm trading occurs, we also examined whether individuals of A. arbustorum adjust the number of sperm they release to the number they receive from their mating partner. There was a high degree of reciprocity in spermatophore transfer: in 45 of the 46 mating pairs investigated both partners delivered a spermatophore that contained spermatozoa. The numbers of sperm transferred by the two mating partners were not correlated, however. This indicates that sperm trading does not occur in this simultaneously hermaphroditic land snail. Copyright 1998 The Association for the Study of Animal Behaviour.