Glucagon-like peptide I reduces postprandial glycemic excursions in IDDM.
Effects of human glucagon-like peptide I (GLP-I)(7-36)amide were examined in volunteers having insulin-dependent diabetes mellitus (IDDM) with residual C-peptide (CP) secretion (n = 8, 7 men and 1 woman; age, 31 +/- 1.4 years; body mass index, 24.7 +/- 0.7 kg/m2; duration of diabetes, 3.2 +/- 0.8 years; insulin dose, 0.41 +/- 0.05 U.kg-1.day-1; meal-stimulated CP, 1.0 +/- 0.2 nmol/l [means +/- SE]). After a mixed meal (Sustacal, 30 kJ/kg body wt), intravenous injection of GLP-I, 1.2 pmol.kg-1.min-1 through 120 min, virtually abolished increments of plasma glucose, CP, pancreatic polypeptide (PP), and glucagon concentrations, with no significant effect on plasma gastrin levels during the infusions. At reduced dosage (0.75 pmol.kg-1.min-1), GLP-I had lesser effects on plasma glucose and CP levels. On cessation of intravenous GLP-I infusions after the meals, plasma glucose, CP, PP, and glucagon concentrations rebounded toward control levels by 180 min, and the response of plasma gastrin was prolonged. These rebound responses are consistent with intestinal delivery of food retained in the stomach on escape from inhibition of gastric emptying by GLP-I. Infusion of 1.2 pmol.kg-1.min-1 GLP-I with 20 g glucose (10% dextrose in water) injected intravenously over 60 min enhanced plasma responses of immunoreactive CP; the mean incremental areas under concentration curves (0-60 min) increased sixfold, but the glycemic excursion was not affected. Thus, in CP-positive IDDM, pharmacological doses of GLP-I reduce glycemic excursions after meals by a mechanism(s) not dependent on stimulation of insulin secretion, presumably involving delayed gastric emptying.(ABSTRACT TRUNCATED AT 250 WORDS)


Classification of β-hairpin repeat proteins.
In recent years, a number of new protein structures that possess tandem repeats have emerged. Many of these proteins are comprised of tandem arrays of β-hairpins. Today, the amount and variety of the data on these β-hairpin repeat (BHR) structures have reached a level that requires detailed analysis and further classification. In this paper, we classified the BHR proteins, compared structures, sequences of repeat motifs, functions and distribution across the major taxonomic kingdoms of life and within organisms. As a result, we identified six different BHR folds in tandem repeat proteins of Class III (elongated structures) and one BHR fold (up-and-down β-barrel) in Class IV ("closed" structures). Our survey reveals the high incidence of the BHR proteins among bacteria and viruses and their possible relationship to the structures of amyloid fibrils. It indicates that BHR folds will be an attractive target for future structural studies, especially in the context of age-related amyloidosis and emerging infectious diseases. This work allowed us to update the RepeatsDB database, which contains annotated tandem repeat protein structures and to construct sequence profiles based on BHR structural alignments.


Cluster microvilli in coronary endothelium.
Examination of cardiac vascular endothelium with scanning electron microscopy, and transmission electron microscopy of previously scanned tissue, revealed several regions of the coronary venous system that contained 'cluster' microvilli. These consisted of 2-15 microvillous projections that emanated radially from a common base or were grouped into a fan-like arrangement. Although rare, these clusters, when present, were widely distributed over the endothelial cell surface.


[Drug-induced icterus].
From the clinical point of view, the classification of drug-induced liver damage into predictable, unpredictable and simulated, has proved useful (Popper and Greim 1973). Forms of jaundice can be predominantly cholestatic or predominantly hepatocellular and can closely imitate an obstructive syndrome or hepatitis. Mixed forms are also frequent.


Studying in the region of proximal learning reduces mind wandering.
Insofar as mind wandering has been linked to poor learning, finding ways to reduce the propensity to mind wander should have implications for improving learning. We investigated the possibility that studying materials at an appropriate level of difficulty with respect to the individual's capabilities-that is, studying in the region of proximal learning (RPL)-might reduce mind wandering. In Experiments 1 and 2, participants were probed for their attentional state while they studied blocks of English-Spanish word pairs that were (a) easy, (b) in the RPL, or (c) difficult. We found that studying materials in the RPL was associated with reduced mind wandering. Test performance on items studied while mind wandering was also poorer. In Experiment 3, we investigated the relation between differences in participants' mastery and mind wandering. We found that high performers mind wandered more when studying the easier word pairs, whereas low performers mind wandered more when studying the difficult items. These results indicate that the RPL is specific to the individual's level of mastery and that mind wandering occurs when people are outside that region.


[Clinical and electroencephalographic signs of side effects in patients after myelography using "amipaque"].
The authors present the results of own investigations on the occurrence of side effects following myelography with the contrast medium Amipaque administered usually by the lumbar route. Sixty patients aged 21-65 years with various diseases of the spinal cord and cauda equina were studied. Clinical and myelographic investigations were performed before and after myelography. In 6 out of 60 cases (10%) transient neurological disturbances developed including epileptic seizures, speech disturbances of the type of aphasia and dysartria, visual disturbances and twitching of lower extremities. Other symptoms and signs included: headaches, vomiting, collapse. One patient with cardiorespiratory failure died hours after myelography with evidence of increased symptoms of cardiorespiratory failure. EEG changes appeared after myelography in 2/3 of cases and persisted for up to 12 days. The authors call attention to the high proportion of neurological complications and EEG changes which must be taken into account when indications to myelography are considered. Particular caution is necessary in cases with coexistent cardiorespiratory failure.


The effects of dihydralazine, labetalol and magnesium sulphate on the isolated, perfused human placental cotyledon.
To assess the effects of dihydralazine, labetalol and magnesium sulphate on the vascular tone in the isolated, perfused human placental cotyledon. In vitro perfusion of the fetal compartment of isolated, human placental cotyledons. None of the drugs affected basal vascular tone. The thromboxane A2-mimic U46619 and endothelin-1 induced a concentration-dependent increment in perfusion pressure, while 5-hydroxytryptamine induced a variable increase, and norepinephrine induced a small, transient increase in perfusion pressure. After preconstriction with U46619, magnesium sulphate (1.5 x 10(-3) to 6 x 10(-3) mol/l) induced a decrease in perfusion pressure, while dihydralazine (10(-6) to 10(-4) mol/l) or labetalol (10(-7) to 10(-4) mol/l) enhanced the perfusion pressure. These effects of dihydralazine and labetalol were unaffected by treatment with indomethacin 10(-6) mol/l, but could be reversed by addition of magnesium sulphate 6 x 10(-3) mol/l. Labetalol 10(-6) to 10(-4) mol/l also caused an increase in the perfusion pressure induced by endothelin-1, but showed no effects after preconstriction with 5-hydroxytryptamine. Pretreatment with labetalol 10(-4) mol/l inhibited the transient increase in perfusion pressure induced by norepinephrine 3 x 10(-5) mol/l. The present data demonstrated that the commonly used vasodilating agents labetalol and dihydralazine do not produce vasodilatation in the human perfused cotyledon after vasoconstriction induced by agents of suggested importance for maintenance of fetal placental vascular tone, and that high concentrations of these drugs may even enhance vasoconstriction induced by thromboxane and endothelin-1 in this area. Magnesium sulphate may show the potential to reverse such unwanted effects of dihydralazine and labetalol.


Activated endothelial cells in culture suppress baroreceptors in the carotid sinus of dog.
Vascular stretch increases the activity of arterial baroreceptors along with the production and release of substances from the endothelium. We hypothesized that endothelial factors modulate the sensitivity of baroreceptors during increases in arterial pressure. Baroreceptor activity was recorded from single fibers innervating the isolated carotid sinus of dogs anesthetized with chloralose after removal of the endothelium (balloon denudation) and after replacing into the denuded sinus bovine aortic endothelial cells cultured on microcarrier beads. The endothelial cells were activated with either the calcium ionophore A23187 (2 microM) or bradykinin (10 microM). The threshold pressure (n = 7) determined with a slow ramp increase in static pressure averaged 73 +/- 7 (SEM) mm Hg during exposure to naked beads and was increased significantly (96 +/- 18 mm Hg; p less than 0.05) during exposure to endothelial cell cultures. During stepwise increases in pressure, activity (n = 6) averaged 14 +/- 5, 40 +/- 8, and 54 +/- 8 spikes/sec at 75, 125, and 175 mm Hg during exposure to naked beads and decreased significantly to 2 +/- 2, 30 +/- 11, and 35 +/- 12 spikes/sec at equivalent pressures during exposure to the cell cultures. The activity was restored after replacement of the cell cultures with naked beads. The suppressed activity was not caused by changes in carotid sinus diameter or strain (sonomicrometers) or by the chemical activators that were also added to the naked beads. The results indicate that chemically activated endothelial cells release an inhibitory factor that suppresses baroreceptor activity.


Nd:YAG laser photodisruption: an experimental investigation on shielding and multiple plasma formation.
In an ocular model plasmas were induced by Q-switched, commercially available Nd:YAG lasers that operated in low order or fundamental mode. Plasma shielding was measured and analysed in relation to energy input, number of plasmas and plasma evolution. Near breakdown threshold, attenuation or shielding of the laser beam resulting from absorption and scattering by the plasma is low and is characterized by high variability from shot to shot. More importantly, shielding does not increase when multiple plasmas are formed, possibly due to a competitive mechanism between these plasmas. Further evidence of this mechanism was obtained from measurement performed using a streak camera. The effectiveness of shielding is scarcely changed by an increase in the cone angle of the incident radiation. In the light of these experiments and other published data, safety guidelines for the clinical use of the Nd:YAG laser have been formulated.


Determination of amino acid tissue concentrations by microdialysis: method evaluation and relation to plasma values.
Microdialysis is an in vivo technique to monitor tissue concentrations of low molecular weight substances by means of a continuously perfused artificial capillary with a semipermeable membrane placed into the region of interest. The suitability of microdialysis to determine tissue concentrations of amino acids was evaluated in vitro by placing the catheter into Ringer buffer or into a plasma protein (50g/l) solution containing 32 different amino acids (150 micromol/l each). All amino acids tested crossed freely the microdialysis membrane with recoveries close to 100%. Microdialysis fluid was sampled from subcutaneous tissue of five newborns and amino acid content analysed. Total and non protein bound amino acids were determined in the patients plasma by acid precipitation or ultrafiltration, respectively. Mean subcutaneous tissue concentrations were lower as compared to plasma for taurine, serine, alanine, aspartate, glutamate and ornithine and higher for valine, isoleucine, leucine, methionine, phenylalanine, tyrosine and arginine, indicating net uptake or release of amino acids from subcutaneous tissue. Thus, microdialysis offers a convenient and minimal invasive way to study tissue amino acid composition and appears to be a promising analytical tool for the study of amino acid metabolism in vivo.


Evaluation of a definition of pre-eclampsia.
To determine: 1. whether an alternative definition of gestational hypertension and pre-eclampsia stratifies women according to their risk of maternal and fetal complications; 2. whether pregnancy outcome in women with gestational hypertension differs in the presence or absence of '+' proteinuria; and 3. whether a blood pressure rise of > or = 30/15 mmHg during pregnancy is associated with adverse outcome in women who remain normotensive. Prospective, nested case-control study. Community based. Healthy, nulliparous women (n = 1496). Women recruited into a study investigating serum markers predictive of pre-eclampsia were classified as having gestational hypertension (systolic blood pressure > or = 140 mmHg with a rise of > or = 30 mmHg and/or diastolic blood pressure > or = 90 mmHg with a rise of > or = 15 mmHg) or pre-eclampsia (gestational hypertension plus proteinuria > or = 2+on dipstick or > 0.3 g/24 h). Maternal and fetal complications in gestational hypertension or pre-eclampsia were compared with a control group of 223 randomly selected normotensive women. The main outcome measures were severe maternal disease, preterm birth and small for gestational age infant. A stepwise increase in adverse maternal and fetal outcomes occurred in gestational hypertension (n = 117, 7.8%) and pre-eclampsia (n = 71, 4.8%). Severe maternal disease developed in 26.5% (21.4% severe hypertension alone, 5.1% multisystem disease) of women with gestational hypertension and 63.4% (21.1% severe hypertension alone, 42.3% multisystem disease) of women with pre-eclampsia (OR 4.8; 95% CI 2.4-9.5). Preterm birth and small for gestational age infants were more frequent in gestational hypertension (OR 1.7; 95% CI 0.5-5.4, and OR 2.0; 95% CI 1.0-3.7, respectively) and pre-eclampsia (OR 14.6; 95% CI 5.8-37.8, and OR 2.6; 95% CI 1.2-5.3) than in the normotensive group. Among women with gestational hypertension severe maternal disease was more common in women with '+' proteinuria (41.7%) than in those with no proteinuria (15.9%): OR 3.8; 95% CI 1.5-9.8. Pregnancies were uncomplicated in the 27% of normotensive women who had a rise of > or = 30 mmHg systolic blood pressure and/or > or = 15 mmHg rise in diastolic blood pressure. In the nulliparous population studied our definition of gestational hypertension and pre-eclampsia identified women at increasing risk of maternal and fetal complications. In gestational hypertension, the presence of proteinuria '+' was associated with a 3.8-fold increase in severe maternal disease. Normotensive women who have a rise in blood pressure > or = 30/15 mmHg had uncomplicated pregnancies.


Efficient magnetization reversal with noisy currents.
We propose to accelerate reversal of the ferromagnetic order parameter in spin valves by electronic noise. By solving the stochastic equations of motion we show that the current-induced magnetization switching time is drastically reduced by a modest level of externally generated current (voltage) noise. This also leads to a significantly lower power consumption for the switching process.


Pneumocystis carinii colonization in the absence of immunosuppression.
A prospective study was undertaken to evaluate the incidence and the course of Pneumocystis carinii colonization in immunocompetent patients with severe pulmonary diseases. A further perspective was to determine the diagnostic values of different detection methods. Bronchoalveolar lavage fluid samples from 77/838 adult HIV-negative patients were examined by Diff-Quik stain, direct immunofluorescence test and polymerase chain reaction. All Diff-Quik stains were negative, but direct immunofluorescence tests and polymerase chain reactions were positive in the samples of 5 patients. The normal number of granulocytes and CD4+T- lymphocytes (median 810 cells/microliters) and normal values of immunoglobulins proved the relative competence of the immune systems of the 77 patients. Although none of these patients received any agent effective against P. carinii, none developed a P. carinii pneumonia within a 120.5-d surveillance period. Nosocomial transmission could be excluded. As the colonization with P. carinii did not result in pneumonia in immunocompetent patients, clinically silent carriers have to be assumed. In non-AIDS patients, sensitive detection methods have to be used to identify colonized persons.


Genetic covariation of neuroticism with monoamine oxidase activity and smoking.
Variation in the personality trait of neuroticism is known to be affected by genetic influences, but despite a number of association studies, the genes involved have not yet been characterized. In a recent study of platelet monoamine oxidase in 1,551 twin subjects, we found a significant association between monoamine oxidase activity and scores on the Eysenck Personality Questionnaire neuroticism scale. Further analyses presented here indicate that both neuroticism and monoamine oxidase activity are associated with variation in smoking habits, and that adjusting for the effect of smoking strengthens the association between MAO and neuroticism. Analysis of the genetic and environmental sources of covariation between neuroticism, smoking, and monoamine oxidase activity show that approximately 8% of the genetic variance in neuroticism is due to the same additive genetic effects that contribute to variation in monoamine oxidase activity, suggesting that variation in neuroticism is associated in part with aspects of serotonin metabolism.


Equilibrium and kinetics of the allosteric transition of GroEL studied by solution X-ray scattering and fluorescence spectroscopy.
We have studied the ATP-induced allosteric structural transition of GroEL using small angle X-ray scattering and fluorescence spectroscopy in combination with a stopped-flow technique. With X-ray scattering one can clearly distinguish the three allosteric states of GroEL, and the kinetics of the transition of GroEL induced by 85 microM ATP have been observed directly by stopped-flow X-ray scattering for the first time. The rate constant has been found to be 3-5s(-1) at 5 degrees C, indicating that this process corresponds to the second phase of the ATP-induced kinetics of tryptophan-inserted GroEL measured by stopped-flow fluorescence. Based on the ATP concentration dependence of the fluorescence kinetics, we conclude that the first phase represents bimolecular non-cooperative binding of ATP to GroEL with a bimolecular rate constant of 5.8 x 10(5)M(-1)s(-1) at 25 degrees C. Considering the electrostatic repulsion between negatively charged GroEL (-18 of the net charge per monomer at pH 7.5) and ATP, the rate constant is consistent with a diffusion-controlled bimolecular process. The ATP-induced fluorescence kinetics (the first and second phases) at various ATP concentrations (< 400 microM) occur before ATP hydrolysis by GroEL takes place and are well explained by a kinetic allosteric model, which is a combination of the conventional transition state theory and the Monod-Wyman-Changeux model, and we have successfully evaluated the equilibrium and kinetic parameters of the allosteric transition, including the binding constant of ATP in the transition state of GroEL.


Antibodies against terminal galactosyl (alpha 1-3) galactose epitopes in systemic sclerosis (scleroderma).
Sera from 224 patients with systemic sclerosis (scleroderma) were analyzed for circulating antibodies against an antigenic determinant characterized by two molecules of galactose in alpha 1-3 linkage. About 45% of the patients were found to have values above the normal range. The mean antibody level was significantly higher than that found in normal subjects (p less than 0.001) or in patients with primary Raynaud's phenomenon who were included as controls. The mean level of anti-Gal antibodies correlated with the degree of skin and internal organ involvement, as well as with the presence of progression or inflammation. Furthermore, when patients with early onset disease were analyzed, high levels of anti-Gal antibodies were present in the subgroups characterized by evidence of progression or inflammation, whereas patients with stable disease did not differ from the controls. We conclude that humoral immunity against Gal alpha 1-3 Gal is an early feature of scleroderma, may be important for its pathogenesis, and may provide a more sensitive tool to detect disease activity.


Preprocessing of Holter ECGs for analysis of the dynamic interrelations between heart rate and ventricular repolarization duration variabilities.
The use of approved commercial digital Holter systems as equipment for data acquisition in cardiologic research presents several potential advantages. In fact, it may provide a standard data acquisition procedure that allows the scientists to concentrate on the data processing phase. As a consequence, it may contribute to the comparability of results among different laboratories. In general, some steps of preprocessing are needed before performing the final analysis on the data. In case of study on the variabilities of the heart rate and the duration of repolarization, preprocessing consists of beat-by-beat measurement time intervals in the 24-hour electrocardiograms. Further preprocessing steps may be required, depending on the particular analysis technique that will be applied to the data. In particular, the present study introduces an operator-free method of preprocessing Holter electrocardiograms for time series analysis of signals related to heart rate variability and variability in the duration of ventricular repolarization.


Follow-Up Results of Newborns after Hearing Screening at a Training and Research Hospital in Turkey.
This study aimed to present the follow-up results of newborns after universal newborn hearing screening at a Training and Research Hospital in İstanbul and to determine the ages of diagnosis, hearing aid fitting, and cochlear implantation in newborns with hearing loss. A total of 5985 newborns were screened between December 2009 and August 2011 using the transient evoked otoacoustic emission test as the first two steps and automated auditory brainstem response (ABR) test as the third step. Newborns who failed the screening tests were referred to a tertiary hospital for clinic ABR and were followed up at least for 2 years. Of 5985 newborns, 5116 (85.5%) completed the screening. Of 53 newborns who were referred to a tertiary hospital, 13 (0.25%) had a hearing impairment. The mean age of diagnosis, hearing aid fitting, and cochlear implantation were 6.1, 9.5, and 24.5 months, respectively. Among the risk factors for hearing impairment, neonatal intensive care (60%) and consanguineous marriage (50%) were the most common ones that were encountered. Our results were consistent with the national literature. Consanguineous marriage may be a risk factor for hearing impairment where it is commonly practiced because consanguineous marriage is significantly high in parents of deaf children. The ages of diagnosis and hearing aid fitting are still beyond the recommended ages by the Joint Committee on Infant Hearing.


Blood cell separation using amphiphilic copolymers containing N,N-dimethylacrylamide.
To develop leukocyte removal filters effective for whole blood, amphiphilic copolymers based on N,N-dimethylacrylamide were synthesized and evaluated as coating materials for poly(ethylene terephthalate) filters. The copolymers with methyl methacrylate or 2-hydroxypropyl methacrylate as a comonomer showed higher platelet permeation ratios (more than 90%) than that of the copolymer with n-butyl methacrylate, though the logarithmic reductions of leukocytes by these copolymers were less than four. An increase in the platelet permeation for whole blood tended to increase the leukocyte permeation. The permeation of both platelets and leukocytes increased with the amount of copolymer coated on the filters because of the change in the physical properties such as the average pore size, total surface area, and total pore volume of coated filters. These results confirm that both the chemical and physical properties of the filters play important roles to control the permeation behavior.


Self-replicating systems.
Over the past 25 years, there has been a surge of development in research towards self-replication and self-replicating systems. The interest in these systems relates to one of the most fundamental questions posed in all fields of science - How did life on earth begin? Investigating how the self-replication process evolved may hold the key to understanding the emergence and evolution of living systems and, ultimately, gain a clear insight into the origin of life on earth. This introductory review aims to highlight the fundamental prerequisites of self-replication along with the important research that has been conducted over the past few decades.


The role of vascular endothelial growth factor leading to vascular leakage in children with dengue virus infection.
Increased vascular permeability is the main aetiology for hypovolaemic shock and circulatory failure in dengue haemorrhagic fever (DHF). To investigate the role of vascular endothelial growth factor (VEGF) in the pathogenesis of DHF. Serum samples from 41 patients [15 dengue fever (DF), 26 DHF] with serologically confirmed dengue virus infection during the febrile, toxic, convalescent stages and at follow-up were analysed for VEGF. Plasma samples from an additional 27 children (16 DF and 11 DHF) during the febrile, toxic stages and at 4-week follow-up and from eight healthy controls were analysed for VEGF. Serum and plasma VEGF levels were not elevated during the febrile or toxic stages of dengue virus infection and did not differ between patients with DF and DHF. Plasma leakage in patients with DHF cannot be explained by elevation of VEGF during the toxic stage of the illness.


Is health workforce sustainability in Australia and New Zealand a realistic policy goal?
This paper assesses what health workforce 'sustainability' might mean for Australia and New Zealand, given the policy direction set out in the World Health Organization draft code on international recruitment of health workers. The governments in both countries have in the past made policy statements about the desirability of health workforce 'self-sufficiency', but OECD data show that both have a high level of dependence on internationally recruited health professionals relative to most other OECD countries. The paper argues that if a target of 'self-sufficiency' or sustainability were to be based on meeting health workforce requirements from home based training, both Australia and New Zealand fall far short of this measure, and continue to be active recruiters. The paper stresses that there is no common agreed definition of what health workforce 'self-sufficiency', or 'sustainability' is in practice, and that without an agreed definition it will be difficult for policy-makers to move the debate on to reaching agreement and possibly setting measurable targets or timelines for achievement. The paper concludes that any policy decisions related to health workforce sustainability will also have to taken in the context of a wider community debate on what is required of a health system and how is it to be funded.


Survival after resection of stage II non-small cell lung cancer.
From 1973 to 1989, 214 patients with stage II non-small cell lung cancer were treated by resection and complete mediastinal lymph node dissection. There were 116 adenocarcinomas and 98 squamous cancers. There were 35 T1 N1 and 179 T2 N1 tumors. Whereas T1 tumors were mainly adenocarcinomas (83%), this difference was not apparent in T2 lesions. Regardless of histology, half of the patients had a single involved N1 lymph node. Lobectomy was performed in 68% of the patients, pneumonectomy in 31%, and wedge resection or segmentectomy in 1%. Lobectomy was sufficient to encompass all disease in 34 of 35 T1 N1 tumors. Only 48 patients (22%) received postoperative external irradiation and 11 patients (5%) received chemotherapy. The overall 5-year disease-free survival was 39%. The best survival rates were in patients who had a single node involved and tumors 3 cm or less in diameter (48%). The pattern of recurrence differed by histology. Local or regional recurrence was more frequent in patients with squamous carcinoma whereas distant metastases were more commonly seen in adenocarcinomas (87%) with brain as the most frequent site (adenocarcinoma, 52%; squamous, 34%). It is concluded that in stage II carcinomas, resection remains the treatment of choice, that mediastinal lymph node dissection provides the most accurate staging, and that the best adjuvant treatment to improve survival is yet to be determined.


Cytochrome P450 and associated monooxygenase activities in the rat and human spinal cord: induction, immunological characterization and immunocytochemical localization.
We have discovered cytochrome P450 and associated monooxygenase activities in microsomes prepared from spinal cord tissues from rats and a human. Cytochrome P450 levels and nicotinamide adenine dinucleotide phosphate cytochrome c reductase activities in microsomes from rat spinal cord were similar to those observed from the whole brain. However, certain monooxygenase activities were significantly lower in the rat spinal cord microsomes as compared to the corresponding activities observed in the whole brain. Cytochrome P450-mediated monooxygenase activities were also detectable in microsomes prepared from human spinal cord. Immunoblot analyses of rat and human spinal cord microsomes using antisera to various forms of hepatic cytochrome P450 namely (2B1 + 2B2), 1A1, 1A2 and 2E1 revealed the presence of immunologically similar forms. The spinal cord microsomes also cross-reacted with the antiserum to the phenobarbital-inducible form of rat brain cytochrome P450. Immunocytochemical stain was predominant in the gray horns of the rat spinal cord. At the cervical level, lamina 1 and 2 representing the substantia gelatinosa were intensely stained. In the ventral horns, lamina 7, 8 and 9 containing the large motor neurons were strongly labelled, while small neurons revealed variable staining. In the white matter, the glial cells were stained but the axons remained non-reactive.(ABSTRACT TRUNCATED AT 250 WORDS)


Synchronization and adaptive control of an array of linearly coupled reaction-diffusion neural networks with hybrid coupling.
In this paper, we propose a general array model of coupled reaction-diffusion neural networks with hybrid coupling, which is composed of spatial diffusion coupling and state coupling. By utilizing the Lyapunov functional method combined with the inequality techniques, a sufficient condition is given to ensure that the proposed network model is synchronized. In addition, when the external disturbances appear in the network, a criterion is obtained to guarantee the H∞ synchronization of the network. Moreover, some adaptive strategies to tune the coupling strengths among network nodes are designed for reaching synchronization and H∞ synchronization. Some criteria for synchronization and H∞ synchronization are derived by using the designed adaptive laws. Numerical simulations are presented finally to demonstrate the effectiveness of the obtained theoretical results.


Growth factor responsiveness declines during adulthood for human skin-derived cells.
Keratinocytes and fibroblasts were obtained from upper arm biopsies of young (22-27 years) and old (60-82 years) adult donors. Keratinocytes were grown in serum-free medium containing variable quantities of either epidermal growth factor (EGF) or a bovine hypothalamic extract known to contain keratinocyte growth factor (KGF). Fibroblasts were grown in serum-free medium containing variable quantities of EGF or insulin. Paired keratinocyte cultures were plated in serum-free medium containing 20% newborn keratinocyte-conditioned medium (NM) or 20% control conditioned medium (CM). Newborn foreskin keratinocytes were plated in 20% conditioned media derived from newborn, young adult or old adult keratinocyte cultures. In spite of large inter-donor variability, keratinocyte growth significantly decreased with age (0.05 greater than P greater than 0.01). Cell yield at 7 days showed an 8-fold increase for young adults over the KGF dose range treated, but only a 4-fold increase for old adults. Young adult cells in varying concentrations of EGF achieved 3-fold to 5-fold higher densities than old, although EGF was not stimulatory for either adult age group. Donor age-associated loss of growth factor responsiveness was confirmed with dermal fibroblasts derived from the same biopsies. Newborn but not adult keratinocytes were stimulated by NM, while newborn cells were not stimulated by either young or old adult conditioned media (YM or OM). An epidermal proliferation index, incorporating both donor cell yield and cell yield of newborn cells in donor conditioned medium, was significantly different (P less than 0.01) for newborn vs. young or old adult cells. Our findings confirm that a decreased proliferative capacity is measurable within adulthood, and suggest that this decrease may be due to a reduced ability to synthesize or respond to mitogens, including autocrine factors.


Hereditary protein C deficiency and portal-vein thrombosis.
Inherited defects of the natural coagulation inhibitors predispose patients to thrombosis. These disorders have similar clinical presentations with a strong family history of thrombosis, episodes of recurrent venous thromboembolism, beginning in early adulthood. We report a case of upper gastrointestinal bleeding in a patient with portal hypertension due to portal-vein thrombosis secondary to hereditary protein C deficiency, an association that has seldom been reported.


[Hepatosplenic sarcoidosis. Apropos of a case].
We present a case of hepatosplenic sarcoidosis without associated pulmonary affection. We have also revised the literature in this respect with special emphasis on the etiopathogenic aspects. The evolution and treatment followed during the period two liver biopsies were performed. The good response to treatment with steroids, clinically and histopathologically, are commented on.


Ethical issues in fetal therapy.
The introduction of routine fetal ultrasound and the technical improvements in ultrasound equipment have greatly increased our ability to diagnose fetal anomalies. As a consequence, congenital anomalies are diagnosed today earlier and in a greater number of patients than ever before. The development of fetal intervention and fetal surgery techniques, improved anesthesia methodology, and sophisticated perinatal care at the limits of viability, have now made prenatal management of some birth defects or fetal malformations a reality. The increasing number of indications for fetal therapy and the apparent desire of parents to seek out these procedures have raised concern regarding the ethical issues related to the therapy. While fetal therapy may have a huge impact on the prenatal management of some congenital birth defects and/or fetal malformations, because of the invasive nature of these procedures, the lack of sufficient data regarding long-term outcomes, and the medical/ethical uncertainties associated with some of these interventions there is cause for concern. This chapter aims to highlight some of the most important ethical considerations pertaining to fetal therapy, and to provide a conceptual ethical framework for a decision-making process to help in the choice of management options.


Opisthorchis viverrini : intensity and rates of infection in cyprinoid fish from an endemic focus in Northeast Thailand.
The infection rates and monthly variations in intensities of Opisthorchis viverrini infection in cyprinoid fish from an endemic focus in Northeast Thailand were investigated between April 1980 and March 1981. Out of six species of cyprinoid fish examined, four were found to harbour O. viverrini metacercariae. All four species of infected fish showed high rates of infection. Among the four species of O. viverrini infected fish, Cycloccheilicthys apagon and Puntius leiacanthus were found in abundance. The results indicate very little variation in the rates of infection throughout the year. On the other hand, intensities of infection varied from month to month, ranging from 8 to 88 metacercariae per fish for C. apagon and from 8 to 32 metacercariae per fish for P. leiacanthus. O. viverrini metacercariae were found in the fish muscle throughout the body; however, the body muscle harboured the highest density of metacercariae, followed by the dorsal fin, pectoral fin, pelvic fin and anal fin muscles respectively. In lieu of all these findings, it is therefore invalid to assume as indicated by the former workers that any potential intermediate fish host that does not have the O. viverrini metacercariae in its pectoral fin muscle in negative with opisthorchiasis.


Probable blind spot in the International Diabetes Federation definition of metabolic syndrome.
The International Diabetes Federation (IDF) proposed a novel definition of the metabolic syndrome (MS) in 2005, which designated central obesity as mandatory. The new National Cholesterol Education Program (NCEP) version, announced by the American Heart Association and National Heart Lung and Blood Institute in October 2005, did not favor any of the five components. We set out to compare the cardiovascular profiles of patients cross-defined by these two definitions to shed light on the differential meanings of the two. We analyzed data from 2608 non-institutionalized adults (> or =19 years old) in the National Nutrition and Health Survey in Taiwan, who had complete data for the five MS defining components. Both definitions adopted lower cut-points for fasting glucose and race-specific cut-points for waist circumference. Under the IDF's and new NCEP's definitions, the MS prevalence was 6.2% and 11.6% in men and 12.6% and 16.5% in women, respectively. Although the two definitions had high agreement, IDF failed to pick up approximately 4% to 5% of people with more than three MS component disorders but a waist circumference less than the cut-point. Subjects whose physical conditions only satisfied NCEP's definition had similar or worse metabolic profiles than those whose conditions satisfied both IDF's definition and the new NCEP's definition. The IDF definition would fail to identify a portion of people who have more than three MS component disorders but a small waistline. Further research and discussion are needed on whether and how to implement the IDF's definition.


Vaccines against dangerous pathogens.
Dangerous pathogens are defined by the UK Health and Safety Executive's advisory committee as category 3 (those which cause severe human disease for which prophylaxis or therapy is usually available) or category 4 (as for category 3, but for which prophylaxis or therapy is not available). Research and development of vaccines for such pathogens is challenging, due to the safety constraints in the manipulation of these pathogens. This chapter discusses the various approaches which can be taken to develop candidate vaccines for these pathogens, including the potential impact of genome sequencing on shortening the time required for R&D. For these pathogens, a direct test of the efficacy of the candidate vaccines in man is not ethical and, therefore, particular emphasis is placed on the demonstration of efficacy in animal models. Emphasis is also placed on the derivation of surrogate markers of efficacy and a demonstration that these correlate with protection in the animal model.


Glycosylation mutants of dengue virus NS1 protein.
The non-structural glycoprotein NS1 of dengue virus type 2 contains sites for N-linked glycosylation at Asn-130 and Asn-207. NS1 synthesized in infected cells is glycosylated at both locations. We have now examined the dimerization and secretion of NS1 lacking one or both of these sites by transient expression of mutagenized cDNA inserted into a simian virus 40-based vector. Immunoblotting and radioimmunoprecipitation were used to detect NS1 associated with transfected cells and in the extracellular medium. Elimination of one or both glycosylation sites did not abolish dimerization and secretion of NS1. However, NS1 lacking Asn-207 showed reduced dimer stability and secretion. Treatment of secreted NS1 with endoglycosidase H demonstrated that complex glycans were attached at Asn-130 and high-mannose glycans at Asn-207.


Spontaneous arrhythmia detected on ambulatory electrocardiographic recording lacks precision in predicting inducibility of ventricular tachycardia during electrophysiologic study.
This study investigates the relation of spontaneous ventricular arrhythmia on ambulatory electrocardiographic (ECG) monitoring to the subsequent inducibility of ventricular tachycardia during programmed electrical stimulation. Eighty patients (65 men, 15 women), whose mean age was 58 years, presented with one of the following: sustained ventricular tachycardia (n = 54); sudden death requiring resuscitation (n = 4); ventricular fibrillation (n = 11); or syncope thought to be of cardiac origin (n = 11). All patients had 24 hour ambulatory electrocardiograms and programmed electrical stimulation while receiving no antiarrhythmic therapy. Programmed electrical stimulation resulted in inducible sustained ventricular tachycardia (defined as a rate of greater than or equal to 120 beats/min for greater than or equal to 1 minute or requiring intervention) in 53 of the 80 patients. There was no measure of frequency or complexity of spontaneous arrhythmia detected on ambulatory ECG that could identify the degree of subsequent ventricular tachycardia inducibility during programmed electrical stimulation. In fact, 25% of patients who had inducible sustained ventricular tachycardia had little or no spontaneous arrhythmia on ambulatory ECG. Furthermore, of the 53 patients with inducible sustained ventricular tachycardia, 28 and 55% had no couplets or nonsustained ventricular tachycardia, respectively, during ambulatory monitoring. The combination of a clinical presentation of sustained ventricular tachycardia, confirmed coronary artery disease and a left ventricular ejection fraction of less than 30% had a better positive predictive value than did any ambulatory ECG criterion in predicting the inducibility of sustained ventricular tachycardia.


Characterized Brillouin scattering in silica optical fiber tapers based on Brillouin optical correlation domain analysis.
This paper demonstrates stimulated Brillouin scattering (SBS) characterization in silica optical fiber tapers drawn from commercial single mode optical fibers by hydrogen flame. They have different waist diameters downscaled from 5 μm to 42 μm. The fully-distributed SBS measurement along the fiber tapers is implemented by Brillouin optical correlation domain analysis technique with millimeter spatial resolution. It is found that the Brillouin frequency shift (BFS) in the waist of all fiber tapers is approximately the same (i.e., ~11.17 GHz at 1550 nm). However, the BFS is gradually reduced and the Brillouin gain decreases from the waist to the untapered zone in each fiber taper.


Anaphylaxis after eating Italian pizza containing buckwheat as the hidden food allergen.
A 20-year-old woman developed anaphylaxis after eating pizza on 4 different occasions in 2 restaurants. Both restaurants made their pizza dough with a mixture of wheat and buckwheat flours. A prick-to-prick test with buckwheat flour was positive. Skin prick tests and specific immunoglobulin E responses to soybean and peanut were weakly positive while the response to buckwheat was negative. We ruled out a pathogenic role for peanut and soybean because the patient usually eats both with no signs of allergic reaction. Double-blind, placebo-controlled food challenges with buckwheat flour were positive after the administration of a cumulative dose of 2.3 g of the culprit flour. To our knowledge, our report describes the first case of anaphylaxis after intake of buckwheat flour as the hidden allergen in pizza dough.


Variations in mineralization affect the stress and strain distributions in cortical and trabecular bone.
The mechanical properties of bone depend largely on its degree and distribution of mineralization. The present study analyzes the effect of an inhomogeneous distribution of mineralization on the stress and strain distributions in the human mandibular condyle during static clenching. A condyle was scanned with a micro-CT scanner to create a finite element model. For every voxel the degree of mineralization (DMB) was determined from the micro-CT scan. The Young's moduli of the elements were calculated from the DMB using constant, linear, and cubic relations, respectively. Stresses, strains, and displacements in cortical and trabecular bone, as well as the condylar deformation (extension along the antero-posterion axis) and compliance were compared. Over 90% of the bone mineral was located in the cortical bone. The DMB showed large variations in both cortical bone (mean: 884, SD: 111 mg/cm(3)) and trabecular bone (mean: 738, SD: 101 mg/cm(3)). Variations of the stresses and the strains were small in cortical bone, but large in trabecular bone. In the cortical bone an inhomogeneous mineral distribution increased the stresses and the strains. In the trabecular bone, however, it decreased the stresses and increased the strains. Furthermore, the condylar compliance remained relatively constant, but the condylar deformation doubled. It was concluded that neglect of the inhomogeneity of the mineral distribution results in a large underestimation of the stresses and strains of possibly more than 50%. The stiffness of trabecular bone strongly influences the condylar deformation. Vice versa, the condylar deformation largely determines the magnitude of the strains in the trabecular bone.


Comparison of non-invasive approaches to red marrow dosimetry for radiolabelled monoclonal antibodies.
Red marrow is usually the dose-limiting organ during radioimmunotherapy. Several non-invasive approaches to calculate the red marrow dose have been proposed. We compared four approaches to analyse the differences in calculated red marrow doses. The data were obtained from immunoscintigraphy of two antibodies with different red marrow kinetics [iodine-131-16.88 IgM and indium-111-OV-TL-3 F(ab')2]. The approaches are based on, respectively, homogeneously distributed activity in the body, a red marrow-blood activity concentration ratio of 0.3, scintigraphic quantification, and a combination of the second and third approaches. This fourth approach may be more adequate because of its independence from the chosen antibody. In addition, the influence of activity accumulation in liver, kidneys or cancellous bone on red marrow dose was studied. The calculated red marrow dose varied between 0.14 and 0.42 mGy/MBq for 111In-OV-TL-3 and between 0.13 and 0.68 mGy/MBq for 131I-16.88. If the radiopharmaceutical shows high affinity for cancellous bone or another organ situated near the red marrow, the activity in these organs must be included in dose calculations. This study shows a large variation in calculated red marrow dose and selection of the definitive non-invasive approach awaits validation.


The essential GTPase RbgA (YlqF) is required for 50S ribosome assembly in Bacillus subtilis.
In this paper the essential GTPase YlqF is shown to participate in the biogenesis of the 50S ribosomal subunit in Bacillus subtilis. Cells depleted of YlqF displayed gene expression profiles and nucleoid morphologies that were consistent with a function for YlqF in translation. In addition, YlqF is evolutionarily linked to two eukaryotic GTPases, Nog2p and Nug1p, that are involved in the biogenesis and the nuclear export of the 60S ribosomal subunit. Analysis of ribosomes from cells depleted of YlqF demonstrated that the formation of 70S ribosomes was greatly reduced and the large subunit sedimented at 45S. Cells grown with varying depleted levels of YlqF, yielding doubling times ranging from 38 min to 150 min, all displayed the 45S intermediate. Purified YlqF-His(6) protein associates with the 45S intermediate, but not the mature 50S subunit in vitro. Analysis of proteins from the 45S intermediate indicated that ribosomal protein L16, which is added late during in vitro Escherichia coli 50S ribosome biogenesis, was missing from the 45S intermediate. These results support a model in which YlqF participates in the formation of active 70S ribosomes in the cell by functioning in a late step of 50S subunit biogenesis. Based on these results we propose to rename the ylqF gene rbgA (ribosome biogenesis GTPase A).


[Association between unplanned pregnancy and adolescence onset asthma].
The association between asthma and pregnancy has been documented previously. The relationship between unplanned pregnancy and onset asthma in adolescence has not been studied. To determine the association between unplanned pregnancy and adolescence onset asthma. A cross-sectional study was done gathering information about asthma, unplanned pregnancy, family atopy and active or passive smoking in 3,130 adolescents aged 13-19 years. Asthma diagnosis was established through a selfquestionnaire based on the International Study of Asthma and Allergy in Childhood. Odds ratios for asthma were determined using logistic regression model and chi-squared test. Mean age of the participants was 16.37 ± 1.93 years. The prevalence of active smoking was 16.1%, of passive smoking 40% and of family atopy 9.8%. From the pregnant adolescents (785), 59.5% reported had planned not to have a child before pregnancy. Prevalence of wheezing during the year prior to the study was 9.3% and of wheezing during the pregnancy 2%. The prevalence of adolescence onset asthma was 5.4%. The age of initiation of asthma in the adolescence was 14.75 ± 1.60 years. The analysis showed that unplanned pregnancy has a slight risk for the development of asthma during adolescence. (Crude OR=1.03; CI 95% 1.02-1.05; p=0.000). Unplanned pregnancy, family atopy, active smoking and smoking friends are associated with the onset-asthma in the adolescence.


Cannabis and driving: a new perspective.
Cannabis and driving is an emerging injury-prevention concern. The incidence of driving while affected by cannabis is rising in parallel with increased cannabis use in the community. Younger drivers are at particular risk. Improvements in research methodology, technology and laboratory testing methods have occurred in the last 10 years. These cast doubt on earlier results and conclusions. Studies now show that cannabis has a significant impairing effect on driving when used alone and that this effect is exaggerated when combined with alcohol. Of particular concern is the presence of cannabis as the sole psychoactive drug in an increasing number of road fatalities and the lack of any structural response to this problem. A review of testing methods, laboratory and real driving studies, and recent epidemiological studies is presented. Suggestions for methods of further data collection and future public policy are made.


Whole genome analysis of an MDR Beijing/W strain of Mycobacterium tuberculosis with large genomic deletions associated with resistance to isoniazid.
Mycobacterium tuberculosis (M.tb) is one of the most prevalent bacterial pathogens in the world. With geographical wide spread and hypervirulence, Beijing/W family is the most successful M.tb lineage. China is a country of high tuberculosis (TB) and high multiple drug-resistant TB (MDR-TB) burden, and the Beijing/W family strains take the largest share of MDR strains. To study the genetic basis of Beijing/W family strains' virulence and drug resistance, we performed the whole genome sequencing of M.tb strain W146, a clinical Beijing/W genotype MDR isolated from Wuxi, Jiangsu province, China. Compared with genome sequence of M.tb strain H37Rv, we found that strain W146 lacks three large fragments and the missing of furA-katG operon confers isoniazid resistance. Besides the missing of furA-katG operon, strain W146 harbored almost all known drug resistance-associated mutations. Comparison analysis of single nucleotide polymorphisms (SNPs) and indels between strain W146 and Beijing/W genotype strains and non-Beijing/W genotype strains revealed that strain W146 possessed some unique mutations, which may be related to drug resistance, transmission and pathogenicity. These findings will help to understand the large sequence polymorphisms (LSPs) and the transmission and drug resistance related genetic characteristics of the Beijing/W genotype of M.tb.


Case report: Neuronal migration disorder associated with chromosome 15q13.3 duplication in a boy with autism and seizures.
Neuronal migration disorders are a group of disorders that cause structural brain abnormalities and varying degrees of neurocognitive impairment, resulting from abnormal neuronal migration during brain development. There are several mutations that have been associated with these disorders. Here the case of a 4-year-old autistic boy is presented, who was found to have evidence of a neuronal migration disorder on magnetic resonance imaging (MRI) during a workup for seizures. Genetic testing did not reveal any of the gene mutations known to be associated with neuronal migration disorders but did reveal a microduplication at chromosome 15q13.3, a locus that has been previously associated with autism, cognitive impairment, and seizures. Although the concurrent presence of the genetic and structural abnormalities does not necessarily imply causality, the simultaneous independent occurrence of both conditions is certainly unusual. It is possible that there may be an association between this duplication syndrome and aberrant neuronal migration.


Complete degradation of butyl benzyl phthalate by a defined bacterial consortium: role of individual isolates in the assimilation pathway.
Two bacterial strains, in consortium, were isolated by enrichment techniques from municipal waste-contaminated soil, which utilized butyl benzyl phthalate (BBP) as the sole carbon source. One of the isolates was identified as Arthrobacter sp. strain WY and the other one as Acinetobacter sp. strain FW based on the morphological, nutritional and biochemical characteristics and 16S rRNA sequence analysis. Various metabolites of BBP engendered by Arthrobacter sp. strain WY were isolated and identified by a combination of chromatographic and spectrophotometric analyses, which revealed a pathway involving monobutylphthalate (MBuP), monobenzyl phthalate (MBzP), phthalic acid and protocatechuic acid. The protocatechuic acid in turn was processed by ortho-cleavage dioxygenase to form beta-carboxy-cis,cis-muconate, ultimately leading to the TCA cycle. The Arthrobacter sp. strain WY could not utilize the hydrolyzed alcohols of BBP. On the other hand, the Acinetobacter sp. strain FW, which by itself could not utilize BBP as the sole carbon source, is capable of utilizing the hydrolyzed alcohols of BBP. Benzyl alcohol was found to be metabolized by the Acinetobacter sp. strain FW via benzaldehyde, benzoic acid and catechol. Catechol was further degraded by ortho-cleavage dioxygenase to cis,cis-muconic acid and subsequently to muconolactone leading to beta-ketoadipate pathway. Moreover, the Acinetobacter sp. strain FW metabolized 1-butanol through butyraldehyde and butyric acid leading to the tricarboxylic acid cycle via beta-oxidation pathway. This is the first report on the complete degradation of BBP by a defined consortium describing the role of its individual constituents in the BBP assimilation pathway.


A review of methods for measuring microplastics in aquatic environments.
An increasing number of reports have been published concerning microplastic (MP) pollution in aquatic environments. Methods used in these studies continue to be updated and lack standardization, so that an up-to-date review pertaining methods for MP research is needed. This critical review examines the analytical methods, including sampling, identification, and quantitation, for MP research. Samples are generally collected from water, sediment, and biota gastrointestinal tract. Manta nets or trawls are prevalently used in surface water sampling, while direct shoveling or box-corer grab are commonly applied in sediment sampling. Microplastics in biota are generally obtained by dissecting organisms and separating livers, gills, and guts. Density separation is frequently chosen to separate MPs from sample matrices. Chemical digestion can dissolve other organic materials and isolate MPs for further identification. Visual sorting should be combined with chemical composition analysis to better identify the polymer type. Pyrolysis or thermal decomposition gas chromatography coupled with mass spectrometry, Fourier transform infrared spectroscopy, and Raman spectroscopy are currently the main technologies for MP identification. Units prevalently used to express MP abundance in water, sediment, and biota are "particles per m3," "particles per m2," and "particles per individual," respectively. As MP abundances often varied with the methods used, we recommend that analytical protocols of MPs should better be standardized and optimized. Despite the important progress in analysis of MPs, detection technologies for identifying nano-sized plastic particles are still lacking, and therefore should be developed swiftly.


[Our evaluation of clinical usefulness of continuous extradural measurement of intracranial pressure in patients with cranio-cerebral injuries].
Continuous measurements of intracranial, pressure were conducted by the extradural method in 40 patients with various types of skull trauma. A correlation was demonstrated between the value of this pressure and the clinical condition of the patient only with high values of the pressure, in which nearly always wave-like pressure changes were observed. The occurrence of wave-like changes was a more reliable sign of threatening cranial hypertension than the value of intracranial pressure. Continuous measurement of pressure during intravenous administration of glycerol demonstrated that a significant fall of pressure preceded the regression of clinical signs of intracranial hypertension.


SCORE risk scale as a prognostic factor after sudden sensorineural hearing loss.
Sudden sensorineural hearing loss (SSHL) is an otologic urgency whose treatment is still controversial. Its etiology remains largely unknown in most cases and predicting its prognosis is still a challenge. Cardiovascular risk factors (CVRF) have been implicated in the etiopathogenesis of this entity. Application of the SCORE (Systematic Coronary Risk Evaluation) risk scale in patients with SSHL and evaluation of its potential prognostic value in recovery in patients with CVRF. Prospective analysis of patients with SSHL admitted for protocol treatment including intravenous corticosteroid therapy associated to weekly intratympanic injection in the event of therapeutic failure or severe hearing loss at admission. Demographic, audiometric, clinical and imaging data were assessed. The SCORE risk scale was applied and the audiometric recovery was compared among different risk groups. Our overall complete and partial recovery rates were 35.9% and 26%, respectively. More than a half of our patients had at least one CVRF. Of these, overweight/obesity, hyperlipidemia and hypertension were the most common. In our sample, patients with CVRF and higher SCORE risk presented higher PTA at admission and also worse hearing outcome, although these results were not statically significant. This preliminary study could not confirm the validity for SCORE scale for cardiovascular risk assessment in predicting audiometric recovery in patients with SSHL with multiple comorbidities. Further research with larger samples are needed to elucidate the etiology of SSHL and the exact role of cardiovascular risk factors in the pathophysiology of SSHL. 4.


Mycoplasmas and 'diphtheroids' in rheumatoid arthritis.
The failure to isolate conventional mycoplasmas from 88 synovial membranes and 119 synovial fluids from patients with proven rheumatoid arthritis using a variety of culture media and techniques agrees with the results of recent workers and suggests that these organisms are unlikely to play a role in the aetiology of the disease. In contrast, 'diphtheroid' organisms were isolated from 27 to 30% of rheumatoid specimens, but not from cultures of non-rheumatoid material. A significantly higher incidence of raised agglutination titres to C. acnes and to a 'diptheroid' isolated from a patient with rheumatoid arthritis in sera from patients with rheumatoid arthritis supports the conclusion that these organisms are present in the joints in such patients, though their role in this disease has yet to be established.


Premorbid adjustment: a phenotype highlighting a distinction rather than an overlap between schizophrenia and bipolar disorder.
Premorbid adjustment (PMA) in schizophrenia (SZ) has been widely studied and shown to be worse in individuals who develop SZ as compared to controls. It has been proposed as a predictor of clinical presentation and outcome, and may delineate a specific SZ phenotype for genetic and other biological studies. Research into PMA in BD has been scarce and inconclusive. The authors compared PMA in individuals suffering from BD with that of healthy controls and investigated whether levels of PMA in BD patients correlate with specific phenotypic features. The authors investigated 344 BD patients and 137 population-based controls. Retrospective PMA assessment was performed using the Premorbid Adjustment Scale (PAS). An overall score as well as sub-scores for age ranges and functional domains were obtained. Patients had a better overall PAS score than controls and outperformed controls during early and late adolescence. They scored significantly better than controls in the functional domains "sociability and withdrawal" and "adaptation to school". No differences were observed for the other subscales and there were no differences between groups during childhood. No association was observed between PMA and any of the phenotype characteristics investigated. In the largest study to date on PMA in BD, PMA was shown to be better in bipolar patients than in healthy controls. PMA in BD is not a simple proxy for commonly studied phenotypic markers of severity. PMA emerges as a phenotype in its own right, and highlights an aspect of disparity rather than overlap between SZ and BD.


[A new direction in the development of Russian silver amalgams. I. The physical research].
A new in principle, ecologically safe and low-waste technology for preparation of an alloy and a formula of several silver alloys containing different levels of silver (OCTA-43, 70 and 80) have been developed. The absence of a alpha-2 phase in amalgams prepared from these alloys is their principally new feature. Physical properties of the new alloys were studied and compared with the known Russian and foreign analogs. Amalgam prepared from CCTA-43 alloy is characterized by optimal characteristics. By their physical characteristics the new compositions are superior to traditional and silver alloys manufactured in Russia at present, and amalgam prepared from OCTA-43 alloy is not inferior to the best foreign analogs.


Infectious hazards in the clinical laboratory: a program to protect laboratory personnel.
The increasing risk of exposure to blood-borne pathogens in the health care setting makes the development of effective infection control programs in the laboratory workplace critical. Central to such programs is the concept of universal precautions. The program described here relates the level of protection or precaution to the potential danger for infection, given the laboratory workstation and task which is to be performed. Four Levels of Protection are described. Implementation of this program requires that each workstation and procedure in each laboratory section be reviewed by the laboratory director and supervisory personnel for risk of exposure. Implementation additionally requires that provisions be made for both the initial and continuing education of laboratory employees. Laboratory directors and supervisors should also monitor the program to ensure compliance. There will certainly be situations unique to individual institutions or laboratory settings that may require precautions or policies over and above those described by universal precautions. Laboratory policies will not gain acceptance if they are developed and implemented without the advice and cooperation of the hospital medical staff. Employee acceptance of infection control policies will be greater if actual development and implementation actively involves the laboratory personnel who will practice them. The program described here is but one approach to the problem. Employers and laboratory directors must understand that it is their responsibility to develop a program that provides appropriate safeguards for workers who may be exposed to infectious agents in the laboratory workplace and to ensure that employees are properly trained and educated in the proper use and application of those safeguards.


Comparison of two forms of long-term potentiation in single hippocampal neurons.
In invertebrate nervous systems, some long-lasting increases in synaptic efficacy result from changes in the presynaptic cell. In the vertebrate nervous system, the best understood long-lasting change in synaptic strength is long-term potentiation (LTP) in the CA1 region of the hippocampus. Here the process is initiated postsynaptically, but the site of the persistent change is unresolved. Single CA3 hippocampal pyramidal cells receive excitatory inputs from associational-commissural fibers and from the mossy fibers of dentate granule cells and both pathways exhibit LTP. Although the induction of associational-commissural LTP requires in the postsynaptic cell N-methyl-D-aspartate (NMDA) receptor activation, membrane depolarization, and a rise in calcium, mossy fiber LTP does not. Paired-pulse facilitation, which is an index of increased transmitter release, is unaltered during associational-commissural LTP but is reduced during mossy fiber LTP. Thus, both the induction and the persistent change may be presynaptic in mossy fiber LTP but not in associational-commissural LTP.


A novel method of preparation of small intestinal brush border membrane vesicles by polyethylene glycol precipitation.
A novel method of brush border membrane vesicle (BBMV) preparation from the small intestinal mucosa using polyethylene glycol (PEG) precipitation has been presented. This preparation is compared with calcium-precipitated BBMVs in marker enzyme enrichment, contamination by other subcellular membranes, transport of glucose, and lipid composition. PEG-precipitated BBMVs are comparable with calcium-precipitated membranes in all parameters except lipid composition and thiol content. PEG-precipitated membranes have more phosphatidylcholine and phosphatidylethanolamine and less lysophosphatidylcholine and lysophosphatidylethanolamine as compared to calcium-precipitated membranes. Diacylglycerol and triacylglycerol content are also high in PEG-precipitated membranes. Alteration in lipid composition indicate the possible activation of lipase and phospholipase by calcium during BBMV preparation, which is not seen in PEG precipitation. Thiol content is almost double in PEG-precipitated membranes as compared to calcium-precipitated membranes. These results indicate that PEG can be used for the preparation of BBMVs in native form from the intestine without any alteration in their structural components, and these membranes show comparable transport activity.


BMAS policy statements in some controversial areas of acupuncture practice.
The safe and competent practice of acupuncture requires the practitioner to conform to a number of basic principles. These include minimising the risk of transmission of infection, maintaining sufficient knowledge of anatomy to guide safe needling, and seeking an orthodox medical diagnosis before embarking on treatment. Beyond these basic principles, there are certain circumstances in practice that generate regular debate amongst practitioners. This article details advice on a limited selection of such circumstances. This advice has been drawn from a set of policy statements originally drafted to facilitate clarity and consistency within the teaching of Western medical acupuncture provided by the BMAS. It is not comprehensive, but covers the areas where there were the greatest differences in approach among the teaching staff. The original policy statements were also incorporated into the BMAS Code of Practice. By using the guidance in this article, in addition to applying sound clinical judgement and a knowledge of relevant anatomy, we hope that practitioners will minimise both the theoretical risks and the reported serious adverse events related to acupuncture, yet be able to practise unencumbered by illogical restrictions.


Quantitative assessment of the interplay between DNA elasticity and cooperative binding of ligands.
Binding of ligands to DNA can be studied by measuring the change of the persistence length of the complex formed, in single-molecule assays. We propose a methodology for persistence length data analysis based on a quenched disorder statistical model and describing the binding isotherm by a Hill-type equation. We obtain an expression for the effective persistence length as a function of the total ligand concentration, which we apply to our data of the DNA-cationic β-cyclodextrin and to the DNA-HU protein data available in the literature, determining the values of the local persistence lengths, the dissociation constant, and the degree of cooperativity for each set of data. In both cases the persistence length behaves nonmonotonically as a function of ligand concentration and based on the results obtained we discuss some physical aspects of the interplay between DNA elasticity and cooperative binding of ligands.


Risk Analysis for Pathological Changes in Pulmonary Parenchyma Based on Lung Computed Tomography Images.
The purpose of this study is to design a content-based medical image retrieval system, which helps excavate and assess pathological change of pulmonary parenchyma for risks analysis. A data set including lung computed tomography images obtained from 115 patients who experienced pathological changes in pulmonary parenchyma is used. Using morphological theory, images are preprocessed and decomposed into groups of pixel blocks (words), which construct vocabulary. A latent Dirichlet allocation (LDA) model is constructed to assess each image for risk analysis with the method of leave-one-out cross-validation. The precision and recall rate are used as the performance assessment criteria. The LDA model generates a relevance rank of retrieval results from high to low. From the top 50 images, precision of identical tissue is 0.76 ± 0.031 and precision of each attribute of pulmonary parenchyma range from 0.776 ± 0.043 to 0.984 ± 0.008. The study results demonstrate that the proposed LDA model is conductive to lung computed tomography image retrieval and has reliable efficacy on risk analysis about pathological changes of pulmonary parenchyma.


Retinotopic order is surprisingly good within cell columns in the cat's lateral suprasylvian cortex.
The retinotopic map in the striate-recipient region of the cat's lateral suprasylvian cortex (referred to here as the lateral suprasylvian area (LS)) has generally been described as quite disorderly. The disorder is commonly attributed to receptive field scatter within cell columns, reflecting the very large size of receptive fields. However, scatter within columns has never been investigated. In the experiments reported here, we examined the receptive field scatter of cells in columns, and also the scatter of a limited sample of their afferents arising from areas 17 and 18. To measure post-synaptic receptive field scatter, electrode penetrations were made parallel to columns in LS, with the electrode approaching from the medial side, traversing the suprasylvian gyrus and emerging into the suprasylvian sulcus. In all 13 such penetrations, receptive fields were clustered together despite their large size. Their centers were scattered over a region that occupied on average less than 20% of the largest field in the column. In contrast, in columns in areas 17 and 18 receptive field centers reportedly are dispersed over regions about equal to the largest of the fields (Hubel and Wiesel 1962, 1965, 1974). The scatter of afferents' receptive fields was assessed anatomically by measuring the overlap between patches of different anterograde tracers in LS. These patches represented terminal labeling from two adjacent or overlapping tracer injections in area 17. While a large degree of overlap would be predicted if afferents have substantial scatter, we found the overlap to be small unless the two injection sites themselves were highly overlapping. Scatter in afferents' receptive fields was measured more directly by physiological recording. In previous experiments, cells in LS were silenced by the local injection of kainic acid, and responses were recorded from axon terminals arising from areas 17 and 18 (Sherk 1989). We examined the receptive field scatter in three penetrations made approximately normal to the cortical surface. Scatter was modest, much less than predicted by the size of post-synaptic receptive fields. Because the degree of receptive field scatter for postsynaptic cells in LS was similar to that of inputs from areas 17 and 18, the scatter of these inputs might be entirely responsible for that seen postsynaptically. Postsynaptic receptive field scatter, on the other hand, was too small to explain the reported disorder in the map in LS.


Pathogen-free screening of bacteria-specific hybridomas for selecting high-quality monoclonal antibodies against pathogen bacteria as illustrated for Legionella pneumophila.
Antibodies are potent biological tools increasingly used as detection, diagnostic and therapeutic reagents. Many technological advances have optimized and facilitated production and screening of monoclonal antibodies. We report here an original method to screen for antibodies targeting biosafety level 2 or 3 pathogens without the fastidious handling inherent to pathogen use. A double ELISA screening was performed using as coated antigen transformed Escherichia coli expressing at its surface a protein specific to the pathogenic bacteria versus control untransformed E. coli. This method was applied to Legionella, using the surface-exposed Mip protein (macrophage infectivity potentiator). This screening proved to be an excellent means of selecting mAbs that bind Legionella pneumophila 1 surface-exposed Mip protein. This method also appears more biologically relevant than screening using the recombinant Mip protein alone and less tedious than a test performed directly on Legionella bacteria. We obtained 21 mAbs that bind strongly to L. pneumophila serogroups 1 to 13, and we validated their use in a rapid ELISA (performed in 4.5 h) and an immunochromatographic test (20 min).


Urogenital paraneurons in several mammals.
Amines and/or peptide-producing cells, deserving to be called paraneurons, were demonstrated in the urethro-prostatic complex of the man, rabbit, buffalo and sheep and in the uterine horns of the pig, horse and mouse, by means of histochemical, immunohistochemical, immunofluorescent and double labeling immunofluorescent techniques. In particular, the urethro-prostatic complex of the sheep contains cells producing serotonin, chromogranin A and somatostatin. Often the amine and the "marker"-protein were colocalized in the same cells. Chromogranin A- and somatostatin-containing cells were found in the uterine horns of the pig. Serotonin probably plays a role in the contraction/relaxation of the musculature of the male urogenital tract, thus regulating the urinary and spermatic flow. The function of somatostatin cells in the urogenital tract is unknown. The presence and direction of their cytoplasmic processes lead us to hypothesize a double endocrine/paracrine modality of action.


Formation of semi-compound C-type starch granule in high-amylose rice developed by antisense RNA inhibition of starch-branching enzyme.
Cereal starch granules with high-amylose and resistant starch (RS) always show irregular morphology and special crystalline structure, but their formation during grain development is not yet clear. In our previous studies, we had generated a transgenic rice line (TRS) enriched with amylose and RS, which contained semi-compound starch showing a C-type crystalline structure. In this study, the formation of semi-compound C-type starch granule during TRS endosperm development was carefully investigated with light, scanning electron, and transmission electron microscopes and X-ray powder diffraction. The results showed that the TRS starch subgranules, each with a central hilum, were individually initiated in amyloplast and showed an A-type crystal at the early stage of starch granule development, which was similar to that in its wild type. However, with the endosperm development, the amylose content in TRS endosperm starch increased and the B-type starch crystal was deposited in the periphery of subgranules; then, the adjacent subgranules fused together and finally formed a continuous outer layer band surrounding the entire circumference of the starch granule. Accordingly, a mechanistic model for the formation of semi-compound C-type starch granules is proposed.


Protective effects of plasma alpha-tocopherols on the risk of inorganic arsenic-related urothelial carcinoma.
Arsenic plays an important role in producing oxidative stress in cultured cells. To investigate the interaction between high oxidative stress and low arsenic methylation capacity on arsenic carcinogenesis, a case-control study was conducted to evaluate the relationship among the indices of oxidative stress, such as urinary 8-hydroxydeoxyquanine (8-OHdG), as well as plasma micronutrients and urinary arsenic profiles on urothelial carcinoma (UC) risk. Urinary 8-OHdG was measured using high-sensitivity enzyme-linked immunosorbent assay kits. The urinary arsenic species were analyzed using high-performance liquid chromatography and hydride generator-atomic absorption spectrometry. Plasma micronutrient levels were analyzed using reversed-phase high-performance liquid chromatography. The present study showed a significant protective effect of plasma alpha-tocopherol on UC risk. Plasma alpha-tocopherol levels were significantly inversely related to urinary total arsenic concentrations and inorganic arsenic percentage (InAs%), and significantly positively related to dimethylarsinic acid percentage (DMA%). There were no correlations between plasma micronutrients and urinary 8-OHdG. Study participants with lower alpha-tocopherol and higher urinary total arsenic, higher InAs%, higher MMA%, and lower DMA% had a higher UC risk than those with higher alpha-tocopherol and lower urinary total arsenic, lower InAs%, lower MMA%, and higher DMA%. These results suggest that plasma alpha-tocopherol might modify the risk of inorganic arsenic-related UC.


[Clinical profiles, pathologic spectrum and management of atrial septal defect in patients aged 50 or over: how should it be treated?].
Clinical and hemodynamic spectrum of secundum type atrial septal defect (ASD) in patients aged 50 or over was studied. Analysis of 20 elderly patients (mean age: 73.6 years) medically treated in Tokyo Metropolitan Geriatric Hospital revealed that in the majority of patients symptoms were mild and shunt flow ratio was relatively small, accompanied by mildly elevated pulmonary arterial pressure. Autopsy studies in six patients suggested small ASD was compatible with normal life span. Analysis of 289 cases aged 50 or over, collected from 42 medical institutions, showed that 26.9% of them were asymptomatic (NYHA functional class I) and 31.3% of them had Qp/Qs ratio of less than 2.0, 20.2% of them had normal or nearly normal pulmonary arterial pressures. Only 16.6% of total cases had been treated surgically. Pulmonary hypertension and congestive heart failure inevitably limited the life span of patients with ASD, so it is clearly reasonable to close the defect before these complications develop. But in view of relatively large proportions of asymptomatic patients and/or patients with normal pulmonary arterial pressure suggestive of small ASD, conservative management may also be appropriate even in those patients aged 50 or over.


Pro-inflammatory and pro-oxidant properties of the HIV protein Tat in a microglial cell line: attenuation by 17 beta-estradiol.
Microglia are activated in humans following infection with human immunodeficiency virus (HIV), and brain inflammation is thought to be involved in neuronal injury and dysfunction during HIV infection. Numerous studies indicate a role for the HIV regulatory protein Tat in HIV-related inflammatory and neurodegenerative processes, although the specific effects of Tat on microglial activation, and the signal transduction mechanisms thereof, have not been elucidated. In the present study, we document the effects of Tat on microglial activation and characterize the signal transduction pathways responsible for Tat's pro-inflammatory effects. Application of Tat to N9 microglial cells increased multiple parameters of microglial activation, including superoxide production, phagocytosis, nitric oxide release and TNF alpha release. Tat also caused activation of both p42/p44 mitogen activated protein kinase (MAPK) and NF kappa B pathways. Inhibitor studies revealed that Tat-induced NF kappa B activation was responsible for increased nitrite release, while MAPK activation mediated superoxide release, TNF alpha release, and phagocytosis. Lastly, pre-treatment of microglial cells with physiological concentrations of 17 beta-estradiol suppressed Tat-mediated microglial activation by interfering with Tat-induced MAPK activation. Together, these data elucidate specific components of the microglial response to Tat and suggest that Tat could contribute to the neuropathology associated with HIV infection through microglial promulgation of oxidative stress.


Complete or a partial sheet of deep temporal fascial graft as a radix graft for radix augmentation.
Autogenous fascia, including the temporalis fascia, has a particular role in rhinoplasty as radix and dorsal onlay grafts. The main purpose of this study is to achieve desired appearance of facial harmony using either a complete or partial sheet of deep temporal fascia as a radix graft in the patients with required permanent radix augmentation. This is a prospective study of 15 patients who referred to Gandi Day Clinic, Tehran, Iran, between 2005 and 2006 and underwent rhinoplasty including nasal radix augmentation using deep temporalis fascia through external rhinoplasty approach. The patients with hump noses had low radix and required radix augmentation. Full sheet facial graft was used in 13 patients. Partial sheet facial graft was used in two patients. As a donor site problem, minor hematoma was observed in one patient. Temporary hair loss in incision line was another problem which were seen in five. As for the recipient site, no infection, persistent erythema of nasal skin extrusion, visibility, displacement of graft materials or irregularity of graft contour were observed postoperatively. Resorption rate was approximately 30% for grafted fascia. Using fascia grafts in radix point, the revision rate was minimal. In order to augment the radix, if 4 mm augmentation is needed, it should be considered that full sheet fascia graft alone would be insufficient. If more than 4 mm of augmentation is needed, 30% overcorrection would manage well the permanent radix augmentation without any problem.


Pemphigus-like antibodies in bullous pemphigoid.
A patient had clinical, histopathologic, and direct immunofluorescent findings compatible with a diagnosis of bullous pemphigoid. In addition, the patient's serum contained pemphigus-like antibodies with different antigenic specificities than true pemphigus antibodies. Absorption of the patient's serum with AB substance failed to block the intercellular substance staining but allowed the demonstration of a basement membrane zone reactive antibody as well. Atypical cases such as this should not cast doubt on the usefulness of immunofluorescence in the diagnosis of bullous skin diseases.


Kidney transplantation in patients suffering from hereditary complete complement C4 deficiency.
Hereditary complete C4 deficiency (C4def) is a very rare condition that predisposes to immune complex disease and end-stage renal failure. Whether such patients should undergo renal transplantation is debated. The clinical outcome of five transplantations in three C4def patients is described. The first patient lost one allograft after 6 years because of chronic allograft rejection. Back on dialysis, he suffered from meningitis caused by Neisseria menigitidis and Aspergillus. One year after a second transplantation under alemtuzumab induction, he developed fulminant Kaposi's sarcoma and died. His sister is now 6 years post-transplantation without complications. The third patient lost his first graft after 3 years because of chronic allograft nephropathy and recurrence of glomerulonephritis. He has now been living with a second graft for over 9 years. He suffered from pneumonia, a generalized varicella infection and Hemophilis parainfluenzae bronchitis. Patients with complete C4def are at increased risk for infection after kidney transplantation. Under certain precautions and with judicious use of immunosuppression, good long-term results are achievable.


Ultimobranchial gland structure in some species of squamata.
Calotes versicolor has single gland on the left side in neck region, just anterior to heart, while Ptyas mucosus and Bungarus caeruleus have 2 equal-sized glands located in the thyro-thymic region. The gland was spheroidal and measured 0.5 to 1.5 mm in diameter. Size of the gland varied individually in relation to the body size. The gland was composed of cell masses and follicles, and was well vascularized. Histology of the gland showed marked individual variation. Ciliated and goblet cells were present.


Cultural competence: a conceptual framework for teaching and learning.
The need to address cultural and ethnic diversity issues in medical education as a means to improve the quality of care for all has been widely emphasised. Cultural competence has been suggested as an instrument with which to deal with diversity issues. However, the implementation of culturally competent curricula appears to be difficult. We believe the development of curricula would profit from a framework that provides a practical translation of abstract educational objectives and that is related to competencies underlying the medical curriculum in general. This paper proposes such a framework. The framework illustrates the following cultural competencies: knowledge of epidemiology and the differential effects of treatment in various ethnic groups; awareness of how culture shapes individual behaviour and thinking; awareness of the social context in which specific ethnic groups live; awareness of one's own prejudices and tendency to stereotype; ability to transfer information in a way the patient can understand and to use external help (e.g. interpreters) when needed, and ability to adapt to new situations flexibly and creatively. The framework indicates important aspects in taking care of an ethnically diverse patient population. It shows that there are more dimensions to delivering high-quality care than merely the cultural. Most cultural competencies emphasise a specific aspect of a generic competency that is of extra importance when dealing with patients from different ethnic groups. We hope our framework contributes to the further development of cultural competency in medical curricula.


Evaluation of MR angiography at 7.0 Tesla MRI using birdcage radio frequency coils with end caps.
The purpose of this study was to evaluate the feasibility of MR angiography (MRA) at 7.0 Tesla (T) using optimized birdcage (BC) coils with simple end cap configurations. Shielded 16-rung high-pass BC coils were built with identical geometry and compared with different sizes and locations of end caps. To determine whether the end cap configuration was effective, the signal intensity profiles along the superior-inferior (S-I) direction were analyzed in phantom and in vivo human experiments. The effects were also investigated in two-dimensional (2D) and three-dimensional (3D) time-of-flight (TOF) MRA experiments. The signal intensity profiles showed that B1 homogeneity at the service end, that is, the end cap side, was improved as the diameter of the end caps increased and the end cap became closer to the coil end ring. The results of 2D and 3D TOF experiments showed the best improvement of vessel visibility at the BC coil with an 80% end cap, when compared with BC coils with other end cap sizes or without an end cap. In conclusion, the BC coil with an end cap was effective for improving S-I directional homogeneity and suitable for MRA applications, especially at ultrahigh field MRI, such as 7.0T.


Voltage-dependent calcium channels.
Voltage-activated calcium channels can be divided into two subgroups based on their activation threshold, low-voltage-activated (LVA) and high-voltage-activated (HVA). Auxiliary subunits of the HVA calcium channels contribute significantly to biophysical properties of the channels. We have cloned and characterized members of two families of auxiliary subunits: alpha2delta and gamma. Two new alpha2delta subunits, alpha2delta-2 and alpha2delta-3, regulate all classes of HVA calcium channels. While the ubiquitous alpha2delta-2 modulates both neuronal and non-neuronal channels with similar efficiency, the alpha2delta-3 subunit regulates Ca(v)2.3 channels more effectively. Furthermore, alpha2delta-2 may modulate the LVA Ca(v)3.1 channel. Four new gamma subunits, gamma-2, gamma-3, gamma-4 and gamma-5, were characterized. The gamma-2 subunit modulated both the non-neuronal Ca(v)1.2 channel and the neuronal Ca(v)2.1 channel. The gamma-4 subunit affected only the Ca(v)2.1 channel. The gamma-5 subunit may be a regulatory subunit of the LVA Ca(v)3.1 channel. The Ca(v)1.2 channel is a major target for treatment of cardiovascular diseases. We have mapped the interaction site for clinically important channel blockers - dihydropyridines (DHPs) - and analysed the underlying inhibition mechanism. High-affinity inhibition is characterized by interaction with inactivated state of the channel. Its structural determinants are amino acids of the IVS6 segment, with smaller contribution of the IS6 segment, which contributes to voltage-dependence of DHP inhibition. Removal of amino acids responsible for the high-affinity inhibition revealed a low-affinity open channel block, in which amino acids of the IIIS5 and IIIS6 segments take part. Experiments with a permanently charged DHP suggested that there is another low-affinity interaction site on the alpha(1) subunit. We have cloned and characterized murine neuronal LVA Ca(v)3.1 channel. The channel has high sensitivity to the organic blocker mibefradil, moderate sensitivity to phenytoin, and low sensitivity to ethosuximide, amiloride and valproat. The channel is insensitive to tetrodotoxin and DHPs. The inorganic blockers Ni2+ and Cd2+ are moderately effective compared to La3+. The current through the Ca(v)3.1 channel inactivates faster with Ba2+ compared to Ca2+. Molecular determinants of fast inactivation are located in amino side of the intracellular carboxy terminus. The voltage dependence of charge movement is very shallow compared to the voltage dependence of current activation. Transfer of 30 % of charge correlates with activation of 70 % of measurable macroscopic current. Prolonged depolarization does not immobilize charge movement of the Ca(v)3.1 channel.


Impaired fetal blood gas status in polyhydramnios and its relation to raised amniotic pressure.
A substantial proportion of perinatal losses in polyhydramnios occur as unexplained normally formed stillbirths. In order to investigate the relationship between fetal condition and raised amniotic pressure (AP), fetal blood gas and acid-base status were determined together with AP in 22 pregnancies with polyhydramnios. At fetal blood sampling, 8 (36%) had a venous pH value and 16 (73%) a pO2 value below the reference range. Both fetal pH and pO2 were significantly negatively correlated with the degree of elevation in AP (y = 7.43 - 0.036x, r = 0.56, p = 0.006, where y = pH and x = AP z score, and y = -1.6 - 0.48x, r = 0.54, p = 0.01, where y = pO2 z score, respectively). Although some of these fetuses were hydropic, had congenital anomalies, or were from multiple pregnancies, univariate and multiple logistic regression analyses indicated that the above associations could not be accounted for by these potentially confounding variables. This work suggests that abnormal fetal blood gas status in human pregnancies with poly-hydramnios is associated with elevated AP.


Identification and characterization of novel low-density lipoprotein receptor mutations of familial hypercholesterolaemia patients in Taiwan.
Familial hypercholesterolaemia (FH) is an autosomal dominant disease associated with a very high risk of coronary vascular disease. The study objective was to identify patients with FH in Taiwan and characterize novel mutations. Fifty-one patients with suspected FH living in Taiwan were screened for mutations in both the low-density lipoprotein (LDL) receptor and the apolipoprotein (apoB) genes using the multiplex polymerase chain reaction and exon-by-exon DNA sequencing technique. Functional consequences on LDL receptor activity were characterized in vitro for novel mutations and family pedigree was also analyzed. Thirteen different functional mutations in the LDL receptor gene and one mutation in the apoB gene were found in 21 patients. Among the 13 mutations in the LDL receptor gene, 10 were single-point missense mutations, one was a two-point mutation in the same allele, one was a non-sense mutation and one was a frame-shift mutation. There were three novel mutations, including two missense mutations (M510K and W512R) and one frame-shift mutation (1953 delTA mutation). The characterization of missense M510K retained 36.2% of the activity of the normal receptor. Conversely, frame-shift 1953 delTA and missense W512R led to defective proteins, with only 0-6% of normal receptor activity. The study identified 13 LDL receptor gene mutations and characterized three novel mutations causing FH in Taiwan. This facilitated a better understanding of FH among the Chinese population and may enable diagnosis of FH at the molecular level at a presymptomatic, early age.


Seroepidemiology of hepatitis B virus, hepatitis D virus, and human immunodeficiency virus infections among parenteral drug abusers in southern Taiwan.
A total of 390 parenteral drug abusers (PDAs) at the Kaohsiung Municipal Narcotics Abstention Institute were examined for markers of hepatitis B virus (HBV), hepatitis D virus (HDV), and human immunodeficiency virus (HIV). All sera were tested for hepatitis B surface antigen (HBsAg), surface antibody (anti-HBs), and core antibody (anti-HBc) by radioimmunoassay (RIA) and for antibody to HIV (anti-HIV) by enzyme-linked immunosorbent assay (ELISA). Hepatitis B e antigen (HBeAg) and antibody to HDV (anti-HDV) were also tested for HBsAg-positive serum samples. Although the HBsAg-positive rate (22.1%) among PDAs was similar to that of the general population in southern Taiwan, the HBV infection rate (99.2%) and the anti-HDV-positive rate (78.5%) among HBsAg-positive subjects were significantly higher than those of the general population in southern Taiwan (P less than 0.0001). None of the PDAs studied were positive for anti-HIV. The levels of serum glutamic oxaloacetic transaminase (SGOT) and serum glutamic pyruvic transaminase (SGPT) among PDAs were significantly higher than those of the general population in southern Taiwan (P less than 0.0001). The more frequent the institutionalisation, the higher the infection rates with HBV and HDV and elevated levels of SGOT and SGPT. Horizontal transmission through parenteral drug abuse may be considered a possible reason for the significantly higher rates of HBV and HDV among parenteral drug abusers.


Semifluorinated alkane based systems for enhanced corneal penetration of poorly soluble drugs.
Semifluorinated alkanes (SFAs) are amphiphilic liquids that can dissolve hydrophobic drugs to form clear solutions. This study evaluated the potential of two SFAs to act as vehicle for topical ocular drug delivery. After confirming ocular safety, an ex vivo corneal penetration model was developed to determine drug distribution and corneal bioavailability. Hydrophobic dye distribution in the different corneal layers was visualised under a confocal microscope. Corneal bioavailability of cyclosporine A (CsA) dissolved in perfluorobutylpentane (F4H5) or perfluorohexyloctane (F6H8) was compared to commercially available CsA ophthalmic emulsions, Restasis® and Ikervis®. Precorneal residence of the four test vehicles containing the hydrophobic dye was also compared using an ex vivo corneal tissue model. Preferential accumulation of the hydrophobic dye in the corneal epithelium was observed with higher amounts detectable when delivered via the SFAs compared to Restasis or Ikervis. A significant improvement in corneal CsA penetration was observed after application of a single dose of 0.05% CsA in F4H5 and F6H8 when compared to Restasis with the area under curve over 4 h (AUC(0-4h)) being at least 8-fold greater for both SFAs (p < .0001). Moreover, the AUC(0-4h) of 0.1% CsA in F4H5 was almost 5-fold greater than Ikervis (p < .0001). Finally, the precorneal residence time of both SFA solutions was significantly longer than that of the commercial emulsions with the AUC(0-60min) being 2- to 11-fold greater. This study demonstrated that SFAs can significantly improve the local bioavailability of hydrophobic drugs by increasing corneal penetration as well as prolonging precorneal residence. They therefore offer a promising new platform for topical drug delivery to the eye.