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Background: Telehealthcare has the potential to provide care for long-term conditions that are increasingly prevalent, such as asthma. We conducted a systematic review of studies of telehealthcare interventions used for the treatment of asthma to determine whether such approaches to care are effective. Methods: We searched the Cochrane Airways Group Specialised Register of Trials, which is derived from systematic searches of bibliographic databases including CENTRAL (the Cochrane Central Register of Controlled Trials), MEDLINE, Embase, CINAHL (Cumulative Index to Nursing and Allied Health Literature) and PsycINFO, as well as other electronic resources. We also searched registers of ongoing and unpublished trials. We were interested in studies that measured the following outcomes: quality of life, number of visits to the emergency department and number of admissions to hospital. Two reviewers identified studies for inclusion in our meta-analysis. We extracted data and used fixedeffect modelling for the meta-analyses. Results: We identified 21 randomized controlled trials for inclusion in our analysis. The methods of telehealthcare intervention these studies investigated were the telephone and video- and Internet-based models of care. Meta-analysis did not show a clinically important improvement in patients’ quality of life, and there was no significant change in the number of visits to the emergency department over 12 months. There was a significant reduction in the number of patients admitted to hospital once or more over 12 months (risk ratio 0.25 [95% confidence interval 0.09 to 0.66]). Interpretation: We found no evidence of a clinically important impact on patients’ quality of life, but telehealthcare interventions do appear to have the potential to reduce the risk of admission to hospital, particularly for patients with severe asthma. Further research is required to clarify the cost-effectiveness of models of care based on telehealthcare.

BackgroundHealthcare systems internationally need to consider new models of care to cater for the increasing numbers of people with asthma. Telehealthcare interventions are increasingly being seen by policymakers as a potential means of delivering asthma care. We defined telehealthcare as being healthcare delivered from a distance, facilitated electronically and involving the exchange of information through the personalised interaction between a healthcare professional using their skills and judgement and the patient providing information.ObjectivesTo assess the effectiveness of telehealthcare interventions in people with asthma.Search strategyWe searched in the following databases: Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, CINAHL, AMED, and PsycINFO; this was supplemented by handsearching of respiratory journals. We also searched registers of ongoing and unpublished trials.Selection criteriaWe selected completed randomised controlled trials of telehealthcare initiatives aiming to improve asthma care.Data collection and analysisTwo review authors independently appraised studies for inclusion and extracted data and performed meta-analyses. We analysed dichotomous variables to produce an odds ratio (OR) and continuous variables to produce a mean difference.Main resultsWe included 21 trials in this review. The 21 included studies investigated a range of technologies aiming to support the provision of care from a distance. These included: telephone (n = 9); video-conferencing (n = 2); Internet (n = 2); other networked communications (n = 6); text Short Messaging Service (n = 1); or a combination of text and Internet (n = 1). Meta-analysis showed that these interventions did not result in clinically important improvements in asthma quality of life (minimum clinically important difference = 0.5): mean difference in Juniper's Asthma Quality of Life Questionnaire (AQLQ) 0.08 (95% CI 0.01 to 0.16). Telehealthcare for asthma resulted in a non-significant increase in the odds of emergency department visits over a 12-month period: OR 1.16 (95% CI 0.52 to 2.58). There was, however, a significant reduction in hospitalisations over a 12-month period: OR 0.21 (95% CI 0.07 to 0.61), the effect being most marked in people with more severe asthma managed predominantly in secondary care settings.Authors' conclusionsTelehealthcare interventions are unlikely to result in clinically relevant improvements in health outcomes in those with relatively mild asthma, but they may have a role in those with more severe disease who are at high risk of hospital admission. Further trials evaluating the effectiveness and cost-effectiveness of a range of telehealthcare interventions are needed.

In this paper, we develop a compositional denotational semantics for prioritized real-time distributed programming languages. One of the interesting features is that it extends the existing compositional theory proposed by Koymanset al (1988) for prioritized real-time languages preserving the compositionality of the semantics. The language permits users to define situations in which an action has priority over another action without the requirement of preassigning priorities to actions for partially ordering the alphabet of actions. These features are part of the languages such as Ada designed specifically keeping in view the needs of real-time embedded systems. Further, the approach does not have the restriction of other approaches such as prioritized internal moves can pre-empt unprioritized actions etc. Our notion of priority in the environment is based on the intuition that a low priority action can proceed only if the high priority action cannot proceed due to lack of the handshaking partner at that point of execution. In other words, if some action is possible corresponding to that environment at some point of execution then the action takes place without unnecessary waiting. The proposed semantic theory provides a clear distinction between the semantic model and the execution model — this has enabled us to fully ensure that there is no unnecessary waiting.

We have obtained 3.5 W of TEM00 output at 532 nm from a longitudinally diode-pumped intracavity-doubled Nd:YAG laser with a diode-to-green conversion efficiency of 14%. Using this laser, we obtained 0.8 W of UV radiation at 266 nm through resonant doubling.

We analyze self-starting in additive-pulse mode-locked lasers and find that dynamic gain saturation can play a determining role. A simple condition for self-starting is presented for comparison with experiments on different laser systems.

A simple model is presented to describe mode locking in a laser coupled to a nonlinear resonator. It reveals a new mechanism for pulse shortening and shows that shortening does not rely on dispersion in the auxiliary cavity. Experimental results are given to support the basic predictions of the model.

In order to characterize the atmospheric optical turbulence at Ali observatory, we have deployed multi-instruments, which are able to continuously monitor the optical turbulence for site evaluation. These instruments include the DIMM, MASS, Single Star SCIDAR and Polaris seeing monitor, and we also plan to install SNODAR and Micro-thermal sensors for the turbulence on surface layer by the end of this year. This configuration allows us to collect a substantial database and make cross-comparison of the results. We have successfully obtained the profiles of optical turbulence and wind speed with Single Star SCIDAR, as well as the key parameters for adaptive optics application, such as seeing, coherence time, and isoplanatic angle. The DIMM seeing measurements are also carried out simultaneously. The median seeing measured by the DIMM and SSS in 2013 is 0.69 and 0.79 arcsec, respectively.

Objectives: To review for acute abdominal pain (AAP), the diagnostic accuracies of combining decision tools (DTs) and doctors aided by DTs compared with those of unaided doctors. Also to evaluate the impact of providing doctors with an AAP DT on patient outcomes, clinical decisions and actions, what factors are likely to determine the usage rates and usability of a DT and the associated costs and likely cost-effectiveness of these DTs in routine use in the UK. Design: Electronic databases were searched up to 1 July 2003. Review methods: Data from each eligible study were extracted. Potential sources of heterogeneity were extracted for both questions. For the accuracy review, meta-analysis was conducted. Among studies comparing diagnostic accuracies of DTs with unaided doctors, error rate ratios provided estimates of the differences between the false-negative and false-positive rates of the DT and unaided doctors' performance. Pooled error rate ratios and 95% confidence intervals (CIs) for false-negative rates and false-positive rates were computed. Metaregression was used to explore heterogeneity. Results: Thirty-two studies from 27 articles, all based in secondary care, were eligible for the review of DT accuracies, while two were eligible for the review of the accuracy of hospital doctors aided by DTs. Sensitivities and specificities for DTs ranged from 53 to 99% and from 30 to 99%, respectively. Those for unaided doctors ranged from 64 to 93% and from 39 to 91%, respectively. Thirteen studies reported false-positive and false-negative rates for both DTs and unaided doctors, enabling a direct comparison of their performance. In random effects meta-analyses, DTs had significantly lower false-positive rates (error rate ratio 0.62, 95% CI 0.46 to 0.83) than unaided doctors. DTs may have higher false-negative rates than unaided doctors (error rate ratio 1.34, 95% CI 0.93 to 1.93). Significant heterogeneity was present. Two studies compared the diagnostic accuracies of doctors aided by DTs to unaided doctors. In a multiarm cluster randomised controlled trial (n = 5193), the diagnostic accuracy of doctors not given access to DTs was not significantly worse (sensitivity 28.4% and specificity 96.0%) than that of three groups of aided doctors (sensitivities of 42.4-47.9%, and specificities of 95.5-96.5%, respectively). In an uncontrolled before-and-after study (n = 1484), the sensitivities and specificities of aided and unaided doctors were 95.5% and 91.5% (p = 0.24) and 78.1% and 86.4% (p < 0.001), respectively. The metaregression of DTs showed that prospective test-set validation at the site of the tool's development was associated with considerably higher diagnostic accuracy than prospective test-set validation at an independent centre [relative diagnostic odds ratio (RDOR) 8.2; 95% CI 3.1 to 14.7]. It also showed that the earlier in the year the study was performed the higher the performance (RDOR 0.88, 0.83 to 0.92), that when developers evaluated their own DT there was better performance than when independent evaluators carried out the study (RDOR = 3.0, 1.3 to 6.8), and that there was no evidence of association between other quality indicators and DT accuracy. The one eligible study of the impact study review, a four-arm cluster randomised trial (n = 5193), showed that hospital admission rates of patients by doctors not allocated to a DT (42.8%) were significantly higher than those by doctors allocated to three combinations of decision support (34.2-38.5%) (p < 0.001). There was no evidence of a difference between perforation rates (p = 0.19) and negative laparotomy rates in the four trial arms (p = 0.46). Usage rates of DTs by doctors in accident and emergency departments ranged from 10 to 77% in the six studies that reported them. Possible determinants of usability include the reasoning method used, the number of items used and the output format. A deterministic cost-effectiveness comparison demonstrated that a paper checklist is likely to be 100-900 times more cost-effective than a computer-based DT under stated assumptions. Conclusions: With their significantly greater specificity and lower false-positive rates than doctors, DTs are potentially useful in confirming a diagnosis of acute appendicitis, but not in ruling it out. The clinical use of well-designed, condition-specific paper or computer-based structured checklists is promising as a way to improve impact on patient outcomes, subject to further research. © Queen's Printer and Controller of HMSO 2006. All rights reserved.

A novel laser tracking measuring system based on the principle of multi-lateration is developed. The system consists of a four-quadrant silicon photoelectric cell measuring the position of reflected beam from the target with sensitivity of 200mv/mm, a rotational tracking mirrror with a solid angle of 35 degrees driven by two motors and a new type of closed-loop control with digital PID and analog PID. The experiment shows that it works reliably with tracking speed 0.7m/s, and the standard deviation of measurement is 1.2μm.

Background: Arterial bifurcations are commonly the sites of developing atherosclerotic plaque that lead to arterial occlusions and plaque rupture (myocardial infarctions and strokes). Laser induced fluorescence (LIF) spectroscopy provides an effective nondestructive method supplying spectral information on extracellular matrix (ECM) protein composition, specifically collagen and elastin. Purpose: To investigate regional differences in the ECM proteins -- collagen I, III and elastin in unstable plaque by analyzing data from laser-induced fluorescence spectroscopy of human carotid endarterectomy specimens. Methods: Gels of ECM protein extracts (elastin, collagen types I & III) were measured as reference spectra and internal thoracic artery segments (extra tissue from bypass surgery) were used as tissue controls. Arterial segments and the endarterectomy specimens (n=21) were cut into 5mm cross-sectional rings. Ten fluorescence spectra per sampling area were then recorded at 5 sites per ring with argon laser excitation (357nm) with a penetration depth of 200 μm. Spectra were normalized to maximum intensity and analyzed using multiple regression analysis. Tissue rings were fixed in formalin (within 3 hours of surgery), sectioned and stained with H&E or Movat's Pentachrome for histological analysis. Spectroscopy data were correlated with immunohistology (staining for elastin, collagen types I, III and IV). Results: Quantitative fluorescence for the thoracic arteries revealed a dominant elastin component on the luminal side -- confirmed with immunohistology and known artery structure. Carotid endarterectomy specimens by comparison had a significant decrease in elastin signature and increased collagen type I and III. Arterial spectra were markedly different between the thoracic and carotid specimens. There was also a significant elevation (p

In his Editorials “Clinical trials and public trust” (3 Dec. 2004, p. [1649][1]) and “The old file-drawer problem” (23 July 2004, p. [451][2]), Donald Kennedy discusses the problem of poor-quality trials funded by pharmaceutical companies in the wake of the Vioxx controversy and the need for

A compact, reliable high power deep ultraviolet (UV) source should encounter much use in areas such as next generation ultra high density optical disk mastering, photo-lithography and material processing. To achieve this goal, an all solid-state approach is imperative. Commercial UV lasers are currently all gas lasers and therefore tend to be bulky and not very efficient. Recently, much work has been reported on UV sources using nonlinear frequency conversion. Previous approaches however were either not all solid-state [1-2] or were characterized by lower UV power [3-5].

OBJECTIVE: To estimate the economic burden of coronary heart disease in the UK using both direct and indirect costs. DESIGN AND SETTING: A prevalence based approach was used to assess coronary heart disease related costs from the societal perspective. PATIENTS: All UK residents in 1999 with coronary heart disease (ICD 9 codes 410-414 and ICD10 codes I20-I25). MAIN OUTCOME MEASURES: Direct health care costs were estimated from spending on prevention, accident and emergency care, hospital care, rehabilitation, and drug treatment. Direct non-health service costs were estimated from data on informal care. "Friction period" adjusted productivity costs were estimated using the human capital approach from lost earnings attributable to coronary heart disease related mortality and morbidity. The friction period is the period of employees' absence from work before the employer replaces them with other workers. Failure to adjust for this factor would overstate production loss. RESULTS: Coronary heart disease cost pound 1.73 billion to the UK health care system in 1999: pound 2.42 billion in informal care and pound 2.91 billion in friction period adjusted productivity loss; 24.1% of production losses were attributable to mortality and 75.9% to morbidity. The total annual cost of all coronary heart disease related burdens was pound 7.06 billion, the highest of all diseases in the UK for which comparable analyses have been done. CONCLUSIONS: Coronary heart disease is a leading public health problem in the UK in terms of the economic burden from disease. Cost estimates would be substantially understated if informal care/productivity costs were excluded.

Initial experiments of a passive polymeric TE-to-TM polarization converter that utilizes laterally offset upper and lower poling electrodes to induce a 45/spl deg/ optical axis in the core of a nonlinear optical polymer waveguide are reported. By optimizing the device length, we have demonstrated polarization conversion efficiencies greater than 95% in devices as short as 240 /spl mu/m with negligible insertion loss.

To elucidate the origination and evolution as well as structure-function relationship of insulin and IGF-1, the gene of amphioxus insulin-like peptide (ILP), a common ancestor of insulin and IGF-1, was chemically synthesized and cloned into the expression vector pVT102-U. In the recombinant ILP, the C-terminus of B-domain and the N-terminus of A-domain of amphioxus ILP deduced from cDNA were linked together by a tripeptide, Ala-Ala-Lys, and its B28Arg residue was also replaced by Lys to facilitate its conversion to a double-chain form by Lys-C cleavage. The expression vector was transformed into yeast cells and the recombinant ILP was expressed efficiently. The purified single-chain and double-chain ILP was obtained by fermentation, purification and enzymatic cleavage. Molecular weight measurement and amino acid composition analysis showed that the primary structure of ILP was correct. Circular dichroism analysis showed that the secondary and tertiary structure of double-chain ILP was similar to that of insulin, while double-chain ILP had no measurable insulin activity in insulin receptor binding assay. Additionally, New Zealand Rabbits were immunized with single-chain ILP and high titre polyclonal antibody was obtained. This work was important for further investigating the molecular evolution of insulin and IGF-1 and the distribution and existence of ILP in amphioxus.

This glossary defines terms used in the comparatively young science of medical informatics. It is hoped that it will be of interest to both novices and professionals in the field.

Buprenorphine (BUP) is an oripavine analgesic that is beneficial in the maintenance treatment of opiate-dependent individuals. Although BUP has been studied extensively, relatively little is known about norbuprenorphine (norBUP), a major dealkylated metabolite of BUP. We now describe the binding of norBUP to opioid and nociceptin/orphanin FQ (ORL1) receptors, and its effects on [(35)S]guanosine-5'-O-(gamma-thio)triphosphate ([(35)S]GTP gamma S) binding mediated by opioid or ORL1 receptors and in the mouse acetic acid writhing test. Chinese hamster ovary cells stably transfected with each receptor were used for receptor binding and [(35)S]GTP gamma S binding. NorBUP exhibited high affinities for mu-, delta-, and kappa-opioid receptors with K(i) values in the nanomolar or subnanomolar range, comparable to those of BUP. NorBUP and BUP had low affinities for the ORL1 receptor with K(i) values in the micromolar range. In the [(35)S]GTP gamma S binding assay, norBUP displayed characteristics distinct from BUP. At the delta-receptor, norBUP was a potent full agonist, yet BUP had no agonist activity and antagonized actions of norBUP and DPDPE. At mu- and kappa-receptors, both norBUP and BUP were potent partial agonists, with norBUP having moderate efficacy and BUP having low efficacy. At the ORL1 receptor, norBUP was a full agonist with low potency, while BUP was a potent partial agonist. In the writhing test, BUP and norBUP both suppressed writhing in an efficacious and dose-dependent manner, giving A(50) values of 0.067 and 0.21 mg/kg, s.c., respectively. These results highlight the similarities and differences between BUP and norBUP, each of which may influence the unique pharmacological profile of BUP.

We propose a novel field‐effect semiconductor laser whose wavelength can be tuned by an electric field parallel to the growth direction of two tightly coupled quantum wells in the active region. We have demonstrated the concept by optically pumping a laser heterostructure whose active region consisted of two 50 A GaAs wells separated by a 20 A Ga0.77Al0.23As potential barrier, in which, at 5 K, we have achieved tuning of the stimulated emission by more than 7 nm.

Previously, we showed that the human kappa-opioid receptor (hkor) stably expressed in Chinese hamster ovary (CHO) cells underwent down-regulation after prolonged U50,488H treatment. In the present study, we determined the mechanisms underlying this process. U50, 488H caused a significant down-regulation of the hkor, although etorphine did not. Neither U50,488H nor etorphine caused down-regulation of the rat kappa-opioid receptor. Thus, similar to internalization, there are agonist and species differences in down-regulation of kappa-opioid receptors. Expression of the dominant negative mutants arrestin-2(319-418) or dynamin I-K44A significantly reduced U50,488H-induced down-regulation of the hkor. Coexpression of GRK2 or GRK2 and arrestin-2 permitted etorphine to induce down-regulation of the hkor, although expression of arrestin-2 or dynamin I alone did not. Expression of the dominant negative mutants rab5A-N133I or rab7-N125I blunted U50,488H-induced down-regulation. Pretreatment with lysosomal enzyme inhibitors [(2S, 3S)trans-epoxysuccinyl-L-leucylamido-3-methylbutane ethyl ester or chloroquine] or proteasome inhibitors (proteasome inhibitor I, MG-132, or lactacystin) decreased the extent of U50,488H-induced down-regulation. A combination of chloroquine and proteasome inhibitor I abolished U50,488H-induced down-regulation. These results indicate that U50,488H-induced down-regulation of the hkor involves GRK-, arrestin-2-, dynamin-, rab5-, and rab7-dependent mechanisms and receptors seem to be trafficked to lysosomes and proteasomes for degradation. Thus, U50,488H-induced internalization and down-regulation of the hkor share initial common mechanisms. To the best of our knowledge, these results represent the first report on the involvement of both rab5 and rab7 in agonist-induced down-regulation of a G protein-coupled receptor. In addition, this study is among the first to show the involvement of proteasomes in agonist-induced down-regulation of a G protein-coupled receptor.

Publication and other forms of selection biases pose a threat to the validity of meta-analysis. Funnel plots are usually used to detect such biases; asymmetrical plots are interpreted to suggest that biases are present. Using 198 published meta-analyses, we demonstrate that the shape of a funnel plot is largely determined by the arbitrary choice of the method to construct the plot. When a different definition of precision and/or effect measure were used, the conclusion about the shape of the plot was altered in 37 (86%) of the 43 meta-analyses with an asymmetrical plot suggesting selection bias. In the absence of a consensus on how the plot should be constructed, asymmetrical funnel plots should be interpreted cautiously. These findings also suggest that the discrepancies between large trials and corresponding meta-analyses and heterogeneity in meta-analyses may also be determined by how they are evaluated.

Opiates active at the mu-opiate receptor (MOR) produce antinociception, in part, through actions involving substance P (SP), a peptide present in both unmyelinated primary afferents and interneurons within the dorsal horn. We examined potential functional sites for interactions between SP and MOR by using dual electron microscopic immunocytochemical localization of antisera against SP and a sequence-specific antipeptide antibody against MOR in rat cervical spinal dorsal horn. The distribution was compared with that of the functionally analogous dorsal horn of the trigeminal nucleus caudalis. Many of the SP-immunoreactive terminals in the dorsal horn contacted dendrites that contain MOR (53% in trigeminal; 70% in cervical spinal cord). Conversely, within the cervical spinal dorsal horn 79% of the MOR-labeled dendrites that received any afferent input were contacted by at least one SP-containing axon or terminal. Although SP-immunoreactive dendrites were rare, many of these (48%) contained MOR, suggesting that the activity of SP-containing spinal interneurons may be regulated by MOR ligands. A few SP-labeled terminals also contained MOR (12% in trigeminal; 6% in cervical spinal cord). These data support the idea that MOR ligands produce antinociception primarily through modulation of postsynaptic second-order nociceptive neurons in the dorsal horns of spinal cord and spinal trigeminal nuclei, some of which contain SP. They also suggest, however, that in each region, MOR agonists can act presynaptically to control the release of SP and/or glutamate from afferent terminals. The post- and presynaptic MOR sites are likely to account for the potency of MOR agonists as analgesics.

Nicotine replacement therapy is probably the most effective of the many methods available to help people to stop smoking (Law & Tang, 1995). Four major forms of nicotine preparations are now available: chewing-gum, patch, inhaler and nasal spray. Chewing-gum and patch have been the most extensively studied in randomized controlled trials, but the four preparations probably have very similar effectiveness: 10–15% of people give up smoking as a result of the treatment (Tang et al., 1994). The effectiveness is highly dependent on the provision of instructions for use and the choice of smokers. Nicotine replacement therapy is also recommend for patients with coronary heart disease and pregnant women since the benefits of treatment well outweigh any harm it may induce. We review the evidence from randomized controlled trials and systematic reviews, raise some issues with regard to the use of nicotine replacement therapy and point out potential ways of increasing the benefit—harm ratio of this therapy.

The role of nucleoside diphosphate kinase (NDKP), which converts GDP to GTP, in the coupling of mu-opioid receptors to G protein was investigated in membranes of Chinese hamster ovary cells stably transfected with the cloned rat mu-opioid receptor (rmor). Endogenous NDPK activity in membranes was determined to be 0.60+/-0.02 micromol/mg protein/30 min UDP (at 10 mM), a competitive substrate of NDPK for GDP with no effect on guanine nucleotide binding to G proteins, reduced basal [35S]GTPgammaS binding and unmasked morphine-stimulated [35S]GTPgammaS binding to pertussis toxin-sensitive G proteins, indicating that [35S]GTPgammaS binding to NDPK accounts for part of its high basal binding. UDP increased the extent of morphine-induced increase in [35S]GTPgammaS binding in the presence of GDP, most likely by reducing basal binding and inhibiting conversion of GDP to GTP. ATP greatly reduced morphine-induced increase in [35S]GTPgammaS binding, whereas AMP-PCP (adenylyl-(beta,gamma-methylene)-diphosphoate tetralithium salt), which cannot serve as the phosphate donor for NDPK, did not, demonstrating that effects of ATP is mediated by the NDPK product GTP. In addition, GDP and ATP increased the Kd and lowered the Bmax of the agonist [3H]DAMGO ([D-Ala2,N-Me-Phe4,Gly5ol]-Enkephalin) for the mu-opioid receptor and GDP alone increased Kd, most likely through their conversion to GTP by NDPK. Addition of exogenous NDPK enhanced the inhibitory effects of GDP and combined GDP and ATP on [3H]DAMGO binding. Thus, NDPK appears to play a role in modulating signal transduction of and agonist binding to mu-opioid receptors.

Objective: The objectives were to estimate the prevalence and correlates of physical abuse-related outcomes in the family setting in Hong Kong’s adolescent population. Method: A cross-sectional study design was used. A randomly selected sample of 3,355 secondary school students in Kwai Tsing District of Hong Kong was surveyed. The response rate was 98%. Results: The prevalence rates of corporal punishment, being beaten by parents for no apparent reason, being beaten to injury by family members in the past 3 months and any one of the above three were 4.9% (95% CI , 4.2% to 5.6%), 2.0% (95% CI , 1.5% to 2.5%), 1.1% (95% CI , .98% to 1.2%) and 6.6% (95% CI , 5.7% to 7.5%), respectively. Students who had experienced the above physical abuse-related outcomes were at a significant disadvantage for a wide range of morbidity indicators, including self-perceived bad health, anxiety and stress, somatic illnesses (such as asthma and epigastric pain), injuries and accidents, and hospitalization. They were more likely to have poor familial relations and coping skills, and to take up habits which potentially put their health at risk, such as smoking, alcohol consumption, and fighting with others. Conclusions: Our prevalence estimates of physical abuse in the family setting for a student population in Hong Kong is an improvement over previous local estimates of physical abuse occurrence, which were mainly based on case notifications and clinical samples. The results also show that the abused adolescents are growing up in an environment filled with physical, psychological, and familial adversities.

To study the mechanism of delaying senility by using the principle of replenishing Qi and promoting blood circulation(RQPBC).The following determinations were carried out in the natural-senile animals which have Qi deficiency and blood stasis Syndrome(24 month old rat) after the treatment. Hemorheological indexes, brain noradrenaline (NE), adrenocorticotropine(ACTH), beta endorphin, cortisol, estradiol, testosterone, interleukin-II (IL-2), tumour necrosis factor (TNF) in the blood, and rat spleen's lymphocyte transformation and induction of IL-2 and the activity of natural killer cells (NKC).The senile rat's above indexes decreased obviously (control: young rats), and by using this principle, all indexes were improved respectively. But these indexes in the young rats cannot be changed by applying this principle.The results indicated that this principle could improve senile body's regulation of neuro-immuno endocrinologic network and keep homeostasis.

Agonist-promoted internalization of some G protein-coupled receptors has been shown to mediate receptor desensitization, resensitization, and down-regulation. In this study, we investigated whether opioids induced internalization of the human and rat kappa opioid receptors stably expressed in Chinese hamster ovary cells, the potential mechanisms involved in this process and its possible role in activation of mitogen-activated protein (MAP) kinase. Exposure of the human kappa receptor to the agonists U50,488H, U69,593, ethylketocyclazocine, or tifluadom, but not etorphine, promoted receptor internalization. However, none of these agonists induced significant internalization of the rat kappa opioid receptor. U50, 488H-induced human kappa receptor internalization was time- and concentration-dependent, with 30-40% of the receptors internalized following a 30-min exposure to 1 microM U50,488H. Agonist removal resulted in the receptors gradually returning to the cell surface over a 60-min period. The antagonist naloxone blocked U50, 488H-induced internalization without affecting internalization itself, while pretreatment with pertussis toxin had no effect on U50, 488H-induced internalization. In contrast, incubation with sucrose (0.4-0.8 M) significantly reduced U50,488H-induced internalization of the kappa receptor. While co-expression of the wild type GRK2, beta-arrestin, or dynamin I had no effect on kappa receptor internalization, co-expression of the dominant negative mutants GRK2-K220R, beta-arrestin (319-418), or dynamin I-K44A significantly inhibited receptor internalization. Whether receptor internalization is critical for MAP kinase activation was next investigated. Co-expression of dominant negative mutants of beta-arrestin or dynamin I, which greatly reduced U50,488H-induced internalization, did not affect MAP kinase activation by the agonist. In addition, etorphine, which did not promote human kappa receptor internalization, was able to fully activate MAP kinase. Moreover, U50,488H or etorphine stimulation of the rat kappa receptor, which did not undergo internalization, also effectively activated MAP kinase. Thus, U50,488H-induced internalization of the human kappa opioid receptor in Chinese hamster ovary cells occurs via a GRK-, beta-arrestin-, and dynamin I-dependent process that likely involves clathrin-coated pits. In addition, internalization of the kappa receptor is not required for activation of MAP kinase.

In this paper the influences of liquid surface vapor injection on various parameters of turbulent boundary layer flow field are given. In the case of low boiling point coolant , such as water , the decrease of liquid surface normal velocity gradient ( 3u/3y) ,. and increase of displacement thickness due to vaporization injection at liquid surface in the region of transpiration nosetip are unlikely to cause a separation of turbulent boundary layer. The correction quantity of pressure distribution caused by the increase of displacement thickness is not big, the approximation of boundary layer P«*P(x) is still tenable. We have given a reasonable definition of displacement thickness of boundary layer under the condition of mass injection.

In this study, we examined whether the human kappa opioid receptor stably expressed in Chinese hamster ovary cells underwent desensitization and down-regulation after prolonged exposure to the agonist (-)U50,488H. Pretreatment with (-)U50,488H led to a reduction in the magnitude of increase in [35S]GTPgammaS binding by the subsequent application of (-)U50,488H. The extent of desensitization was related to duration of exposure and (-)U50,488H concentration. Pretreatment with (-)U50,488H also reduced the potency of (-)U50,488H in inhibiting forskolin-stimulated adenylate cyclase. In membranes of (-)U50,488H-pretreated cells, the affinity of (-)U50,488H was lower than that in the untreated control, and GTPgammaS had no effect on (-)U50,488H affinity, consistent with the notion of uncoupling of the receptor-G protein complex by (-)U50, 488H treatment. Down-regulation of the kappa opioid receptor also occurred on exposure to (-)U50,488H. Higher (-)U50,488H concentrations and/or longer incubation periods were required for down-regulation than for desensitization. The degree of down-regulation depended on the agonist concentration and incubation time. (-)U50,488H-induced desensitization and down-regulation were blocked by naloxone. (+)U50,488H, an inactive stereoisomer, did not cause desensitization or down-regulation. These results indicate that both processes were receptor-mediated. After incubation with (-)U50,488H and removal of (-)U50,488H, both (-)U50,488H-induced [35S]GTPgammaS binding and receptor number returned to the control level, which indicates that both processes were reversible. Thus, desensitization and down-regulation of the kappa opioid receptor occur after agonist exposure and represent two different adaptation mechanisms.

This study is a retrospective analysis of the clinical data of 67 very low birth weight infants (VLBWI) with symptomatic patent ductus arteriosus (PDA); all were admitted to four neonatal intensive care units (NICU) from January 1, 1996 through December 31, 1996. The mean gestational age was 27.9 +/- 2.4 weeks, the mean birth weight was 1078 +/- 193 g. Fifty-six infants (83.6%) had RDS, and 53 infants (79%) received artificial surfactant. The NICU at a regional hospital used CVD scoreor =3 as the sole criteria and the remaining three NICUs used color Doppler echocardiogram to confirm a symptomatic PDA and to treat it; sometimes LA/AOor = 1.3 was used as the criteria for indomethacin treatment. Two of these three hospitals sometimes used the pulsed Doppler echocardiogram as well as color Doppler examination as the treatment criteria. Seven infants (10.5%) had contraindication for indomethacin treatment; four of them closed after conservative treatment, and another three were subjected to surgical ligation. Of the remaining 60 infants, 3 were treated with oral indomethacin and 57 were treated with intravenous indomethacin. The mean age when initial treatment given was 3.8 +/- 1.5 days (range, 8 hours approximately 20 days). Among them 10 (16.7%) were within 24 hours after birth, 25 (41.7%) were between 24 and 48 hours, and 25 (41.7%) were beyond 48 hours. The dosage of indomethacin was 0.2 mg/kg per dose intravenously every 12 to 24 hours for three doses as a full course, if not contraindicated. The mean dose of indomethacin was 2.8 +/- 1.5; 10 infants (16.7%) received 1 dose, 15 (25%) received 2 doses, 27 (45%) received 3 doses, 3 (5%) received 4 doses and 5 (8.3%) received 6 doses. Among them, 51 infants (85%) PDA closed (including 2 treated with oral indomethacin), 9 (15%) failed to close and 6 of them received surgical ligation (including 1 treated with oral indomethacin). The complications associated with indomethacin treatment were hypoglycemia (52%), decreased urine output (42%) and gastrointestinal hemorrhage (32%). The infants with RDS had an earlier mean age of initial treatment than non-RDS infants (3.3 +/- 2.5 vs. 7.6 +/- 5.6 days; p0.05), and also had a higher closure rate (89% vs. 57%; p0.05). There was a closure rate of 85% in this multicenter retrospective analysis. Even though the infants received only one or two doses, they still had a good chance of ductal closure (21/25, 84%). To minimize the complications associated with indomethacin treatment in VLBWI, the protocol of indomethacin treatment should be re-evaluated.

Mutational analysis of opioid receptors supports the hypothesis that dissimilar receptor domains contribute to the binding affinity of different ligands. To determine whether enantiomeric ligands can serve to distinguish between different binding pockets (which focuses the analysis on asymmetric structural factors while avoiding confounding changes in physiochemical characteristics), we analyzed the binding of the 3-methylfentanyl congeners RTI-4614-4 [(+/-)-cis-N-[1-(2-hydroxy-2-phenylethyl)-3-methyl-4-piperidyl]-N- phenylpropanamide HCl)], its four stereoisomers [(2S,3R,4S)-1a, (2R,3R,4S)-1b, (2R,3S,4R)-1c, and (2S,3S,4R)-1d], and other mu agonists with cloned rat mu opioid receptors stably expressed in HEK-293 cells and mu/kappa receptor chimeras. Chimera III (kappa[aminoacids 1-141]/mu[aminoacids 151-398]), chimera IV (mu[aminoacids 1-150]/kappa[aminoacids 142-380]), and chimera XII (kappa[aminoacids 1-262]/mu[aminoacids 269-398]) bound [(125)I]IOXY (6beta-iodo-3,14-dihydoxy-17-cyclopropylmethyl-4,5alpha++ +-epoxymorphinan) with high affinities. The Ki values of 1a, 1b, 1c, and 1d at the wild-type mu receptor were 0.55 nM, 0.66 nM, 124 nM, and 59.2 nM, respectively. When the region from the N terminal to the start of the transmembrane helix 3 (TMH3) of the mu receptor was substituted by that of the kappa receptor (chimera III), the Ki value of 1b was increased (relative to the mu receptor) 590-fold compared to a 73-fold increase for 1a. When this portion of the kappa receptor was replaced by that of the mu receptor (chimera IV), the loss of affinity was not as great: 11.7-fold for 1a and 58.5-fold for 1b. Replacement of the middle of the third intracellular loop and third extracellular loop (e3) of the kappa receptor with that of the mu receptor (chimera XII) lowered (relative to their Ki values at the kappa receptor) the Ki values of [D-Ala2,D-Leu5]enkephalin and [D-Ala2-MePhe4,Gly-ol5]enkephalin to a much greater extent than the Ki values of the isomers. The kappa/chimera XII shift was greater for isomers 1c and 1d than for 1b and 1a. Viewed collectively, these data suggest that the region from the N terminal to the start of the TMH3 of the mu opioid receptor determines the binding affinity of RTI-4614-4 and its isomers and that the e3 loop also plays a major role in determining the binding affinity of mu agonist peptides. These data also show that the stereoisomers of RTI-4614-4 probably bind to different domains of the mu receptor and suggest that manipulation of stereochemistry may be a useful tool for designing domain-specific ligands.

Intracerebral microdialysis was used to measure changes in the extracellular level of substance P (SP) released from the periaqueductal gray (PAG) and the preoptic anterior hypothalamus (POAH) of freely moving Sprague-Dawley rats after noxious cold stimulation. Artificial cerebrospinal fluid was perfused into the dialysis probe in the PAG or POAH and samples were collected every 30 min for 4 hr. SP-like immunoreactivity in the samples was measured by radioimmunoassay. In the PAG, SP base-line release was 0.43 +/- 0.08 fmol/fraction. SP release was increased to 1.3 +/- 0.4 fmol/fraction during the first collection period after noxious cold. Pretreatment with the selective mu opioid receptor agonist PL017 (0.8-3.4 nmol) or the kappa opioid receptor agonist dynorphin A1-17 (4.6-9.2 nmol), administered into the PAG by microinjection, produced dose-related inhibition of the cold-evoked SP release. Naloxone (10 mg/kg s.c.) administration 10 min before these opioid agonists reduced the inhibition of SP release. In the POAH, SP base-line release was 0.45 +/- 0.06 fmol/fraction and noxious cold did not cause any significant change in SP release. Microdialysis of SP (271 fmol-271 pmol/microl/min, for 30 min) into the PAG, but not the POAH, induced dose-related analgesia (35-68% MPA) in the cold-water tail-flick test. However, microdialysis of SP into the POAH or PAG failed to induce any significant change in body temperature. These data suggest that 1) SP released from the PAG acts as a neuromodulator to transmit nociceptive information; 2) opioid receptor agonists can suppress this information by inhibiting SP release; 3) SP evoked by noxious cold may have a role in triggering the antinociceptive function of the PAG; and 4) SP does not appear to act as a neuromodulator for thermoregulatory responses in the POAH.

Activation of kappa receptors inhibits adenylate cyclase, enhances K+ conductance and reduces Ca++ conductance via pertussis toxin-sensitive G proteins. We recently cloned a human kappa opioid receptor and stably expressed it in Chinese hamster ovary (CHO) cells. In this study, the effects of activation of the human kappa receptor by agonists on [35S]GTPgammaS binding to CHO cell membranes were examined. The presence of GDP and Mg++ was essential for the kappa agonist (-)-U50,488H-induced increase in [35S]GTPgammaS binding to be observed and the optimal concentration was 3 microM and 5 mM, respectively. The presence of 100 mM Na+ was necessary to produce the maximal signal-to-background ratio. (-)U50,488H-induced increase in [35S]GTPgammaS binding was time- and tissue concentration-dependent. (-)U50,488H increased [35S]GTPgammaS binding in a dose-dependent manner with an EC50 of 3.1 nM. (+)-U50,488H had no effect, which indicates that this effect is stereospecific. Naloxone (1 microM) or norbinaltorphimine (10 nM) shifted the dose-response curve of (-)-U50,488H to the right by 100-fold. These results indicate that enhancement of [35S]GTPgammaS binding by (-)-U50,488H is a kappa receptor-mediated event. Pretreatment of the cells with pertussis toxin, but not cholera toxin, abolished the (-)-U50,488H-induced increase in [35S]GTPgammaS binding, which indicates the involvement of Gi and/or Go proteins. [35S]GTPgammaS binding induced by (-)-U50,488H had a Kd value of 0.34 +/- 0.08 nM and a Bmax value of 431 +/- 29 fmol/mg protein. The rank order of potencies of opioid ligands tested in stimulating [35S]GTPgammaS binding was dynorphin A 1-17(+/-)-ethylketocyclazocinebeta-funaltrexamine, (-)-U50,488H, tifluadomnalorphinepentazocine, nalbuphinebuprenorphine. Dynorphin A 1-17, (+/-)-ethylketocyclazocine, (-)-U50,488H, tifluadom and beta-funaltrexamine were full agonists, but nalorphine and pentazocine were partial agonists producing maximal responses of 68% and 23% of those of full agonists, respectively. Nalbuphine and buprenorphine had low levels of agonist activities. Norbinaltorphimine and naloxone were antagonists devoid of activities. Enhancement of [35S]GTPgammaS binding by kappa agonists provides a simple functional measure for receptor activation and can be used for determination of potencies and efficacies of opioid ligands at the kappa receptor.

A new fluorescence polarization (FP) immunoassay was developed for the COBAS INTEGRA to quantitate lidocaine in serum and sodium heparin derived plasma. The COBAS INTEGRA, a random and continuous access clinical analyzer, performs the new lidocaine assay at a throughput of 300 tests per hour. Comparison studies with the Abbott TDx Lidocaine assay yielded a correlation coefficient of 0.99 and a regression value of INTEGRA = -0.01 + 0.95 TDx, n = 69. The assay was found to be linear throughout the concentration range of 0 to 10 micrograms/ml using CASCO STANDARDS's DOCUMENT TDM 1 Linearity Test Set and the COBAS-FP TDM Calibration Verification Test Set. The new lidocaine assay exhibited good precision as determined by the National Committee for Clinical Laboratory Standards (NCCLS) protocol EP5-T2 with total percent coefficient of variation (%CV) values of less than 4 percent across the assay range. Interference was less than 10 percent for abnormal levels of total protein (2.1 to 10 g/dl), lipid (up to 2200 mg/dl triglyceride), hemoglobin (up to 10 g/dl), and icteric samples (up to 17.5 mg/dl bilirubin). Finally, the standard curve was stable for greater than 25 weeks which was attributable to the on-board reagent cooling and a unique cassette closure system.

The practice of Traditional Chinese Medicine (TCM) is largely unregulated in Hong Kong. Yet, as previous studies have shown, a sizable segment of the population consults TCM practitioners for health problems. This paper uses health care utilization data from a telephone health survey of 847 adult subjects in Tai Po District who had suffered from acute illness in the past month, to examine the profile of TCM users in the District. Women, older residents, unemployed workers, low skill laborers, current smokers and subjects dissatisfied with the quality of private sector clinics were significantly more likely to consult TCM practitioners.

Initial experiments of a passive polymeric TE-to-TM polarization converter that utilizes laterally offset upper and lower poling electrodes to induce a 45° optical axis in the core of a nonlinear optical polymer waveguide are reported. By optimizing the device length, we have demonstrated polarization conversion efficiencies greater than 95% in devices as short as 240 um with negligible insertion loss.

Activation of the mu opioid receptor (MOR) by morphine within the caudal nucleus of the solitary tract (NTS) is known to mediate both cardiorespiratory and gastrointestinal responses. Leu5-enkephalin (LE), a potential endogenous ligand for MOR, is also present within neurons in this region. To determine the cellular sites for the visceral effects of MOR ligands, including LE, we used immunogold-silver and immunoperoxidase methods for light and electron microscopic localization of antisera against MOR (carboxyl terminal domain) and LE in the caudal NTS of rat brain. Light microscopy of coronal sections through the NTS at the level of the area postrema showed MOR-like immunoreactivity (MOR-LI) and LE labeling in punctate processes located within the subpostremal, dorsomedial and medial subnuclei. Electron microscopy of sections through the medial NTS at this level showed gold-silver particles identifying MOR-LI prominently distributed to the cytoplasmic side of the plasma membranes of axons and terminals. MOR labeled terminals formed mostly symmetric (inhibitory-type) synapses but sometimes showed multiple asymmetric junctions, characteristic of excitatory visceral afferents. MOR-LI was also present along extrasynaptic plasma membranes of dendrites receiving afferent input from unlabeled and LE-labeled terminals. We conclude that MOR ligands, possibly including LE, can act at extrasynaptic MORs on the plasma membranes of axons and dendrites in the caudal NTS to modulate the presynaptic release and postsynaptic responses of neurons. These are likely to include local inhibitory neurons and both gastric and cardiorespiratory afferents known to terminate in the subnuclei with the most intense MOR-LI.

Simplified analysis for thermal protection of the self-contatined Adaptive Transpiration (SCAT) nosetip is provided and the simplified calculation formulas of thermal protection are deduced. The calculation examples of thallium, indium, tin and water coolants are given. The deduced formulae in this paper are simple and convenient. An engineering calculation method of nosetip thermal protection design is obtained in this paper. It can be used in prediction before the ground simulation experiment.

We describe a 0.4W average power at maximum, frequency-quintupled Q-switched Nd:YAG laser at a repetition rate of 7 kHz, which is a potential light source for next generation microlithography. Calculated results for the conversion efficiencies considering pump depletion will be discussed. Our results allow to foresee further scaling up 213 nm power up to the 1W level by increasing the fundamental power.© (1996) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

High power deep ultraviolet (UV) radiation has attracted much attention in areas such as photo-lithography, micro fabrication, material processing and ultra high density optical disk mastering. We report progress in improving the quality of Czochralski-grown (beta) -BaB2O4 (BBO) which is essential for generating high average output power in the deep ultraviolet regime. We obtained 1.5 W of cw 266 nm output using an external resonant doubler, 1.3 W of 213 nm output from a flashlamp pumped Q-switched Nd:YAG laser and more than 0.4 W of 213 nm radiation from a commercial 6 W diode-pumped high repetition rate Q-switched Nd:YAG laser using our melt-grown BBO crystals for the nonlinear frequency conversion. Using the cw 266 nm output speckle-free 0.25 micrometers photolithography and micro-fabrication was demonstrated.© (1996) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

Although laser angioplasty has been demonstrated to be effective for the treatment of long, complex coronary arterial atherosclerotic stenoses, there is an associated risk of acute arterial spasm, dissection, and perforation as well as a significant restenosis rate. It has been postulated that the use of lasers emitting at wavelengths designed for radiation absorption by water would decrease local tissue trauma.We have examined the use of a Nd:YAG laser designed to emit at 1.44 microns, an absorption peak for water, and compared the results of laser ablation at 1.06 microns, 1.32 microns, and 1.44 microns wavelengths. Nd:YAG laser angioplasty was performed in the abdominal aorta of White Leghorn roosters. Acute and chronic vascular trauma was assessed by contrast angiography and histological analysis.There was a significant decrease in early mortality with 1.44 microns laser ablation. This decreased mortality after 1.44 microns ablation was associated with a decrease in vascular spasm, perforation, and thermal damage. Atherosclerotic plaque development at follow up was decreased with 1.44 microns ablation but this was not significant.1.44 microns laser ablation decreases early vascular trauma and mortality and may decrease subsequent atherosclerotic plaque development.

To investigate the interaction between the mu opioid receptor and its ligands, we compared the binding of mu-selective ligands to two mu/kappa chimeric opioid receptors and to mu and kappa receptors. The two chimeras were constructed from cloned rat mu and kappa receptors in which a segment from the middle of the third intracellular loop to the C terminus was exchanged. When this portion of the kappa receptor was replaced by that of the mu receptor, affinities of mu selective agonists, DAMGO (Tyr-D-Ala-Gly-NMePhe-Gly-ol), PL017 (Tyr-Pro-NMePhe-D-Pro-NH2), sufentanil, and morphine, were greatly increased as compared to those for the kappa receptor. Conversely, when this region of the mu receptor was substituted by that of the kappa receptor, affinities for these agonists were substantially decreased as compared with those of the mu receptor. Unlike selective agonists, the mu-selective antagonist, CTAP (D-Phe-Cys-Tyr-D-Trp-Arg-Thr-penicillamine-Thr-NH2), displayed a low affinity for both chimeric receptors, similar to that of the kappa receptor. Thus, the region from the middle of the third intracellular loop to the C terminus of the mu receptor is important for the binding of selective agonists. Conversely, the determinants for selective binding of the antagonist CTAP reside in a more extended region of the receptor.

Effects of 24 h pretreatment with intracerebroventricular (icv) beta-funaltrexamine (beta-FNA) on brain opioid receptor binding in rats were examined under various conditions. Agonist binding to mu and delta opioid receptors (with [3H][[cap]dAla2,MePhe4,Gly-ol5]enkephalin (DAMGO)[3H][D-Pen2, D-Pen5]enkephalin (DPDPE), respectively) was performed under three different conditions: i) pretreatment of membranes with GDP and Na+ and binding in the presence of Mg++ in Tris-HCI buffer containing EGTA and leupeptin for 1.5 to 3 h; ii) binding in Tris-HCI buffer containing bacitracin, leupeptin, chymostatin and bestatin for 3 to 4 h; iii) binding in Tris-HCI buffer containing EGTA and leupeptin for 45 min. Condition i was shown to convert opioid receptors to a high affinity state for agonists. beta-FNA (2, 6 or 20 nmol) significantly reduced 1 nM [3H]DAMGO binding in the whole brain with i but not with ii. With iii, 20 nmol beta-FNA reduced [3H]DAMGO binding, but not 2 or 6 nmol. Saturation experiments with i showed that the reduction in [3H]DAMGO binding after 6 or 20 nmol beta-FNA was due to a decrease in Bmax and an increase in KD. For delta binding, there was no significant change in [3H]DPDPE (2 nM) binding with i after 2, 6 or 20 nmol beta-FNA. Thus, under i, icv beta-FNA reduced [3H]DAMGO binding significantly without affecting [3H]DPDPE binding. In addition, mu binding was also conducted with 1 nM [3H]naloxone under three different conditions: iv) in the presence of Na+ and GDP; v), in the presence of Na+, Gpp(NH)p and Mg++; vi) in the presence of Na+. Both iv and v were shown to shift opioid receptors to a low affinity state for agonists. beta-FNA (20 nmol) significantly decreased 1 nM [3H]naloxone binding under each of the three conditions. Competitive inhibition of 1 nM [3H]naloxone binding by DAMGO in the presence of Na+ and GDP showed that receptors existed in a single low affinity state for DAMGO, and that icv beta-FNA caused a reduction in Bmax without affecting the KD of DAMGO. In summary, when all the receptors were converted to a high agonist affinity state i or a low agonist affinity state iv, the changes in mu binding induced by beta-FNA could be revealed with agonist binding. Additionally, changes in mu binding induced by beta-FNA could be detected with [3H]naloxone, which always displayed high affinity regardless of agonist affinity states, under each of the three conditions (iv, v and vi).

The R1.1 cell line has been shown to express kappa-opioid receptors on the cell surface. Our analysis shows that the R1.1 cell line exhibits a CD4NEG CD8NEG CD3LOW CD25LOW cell surface phenotype, characteristic of thymocytes in one of the early stages of differentiation. We have developed reverse transcriptase polymerase chain reaction (RT-PCR) conditions that permit the detection of mRNA coding for the kappa-receptor. Using cell fractionation techniques we have isolated CD4NEG CD8NEG thymocytes, and analysis by RT-PCR shows that these primary immature thymocytes also express the kappa-opioid receptor. We hypothesize that the expression of kappa-opioid receptor may be a marker which is characteristic of immature T development.

This study was to identify specific regions in kappa opioid receptors that accounted for binding selectivity of kappa ligands. Six chimeric mu/kappa receptors were constructed from cloned rat kappa and mu opioid receptors and transiently expressed in COS-1 cells. All six chimeric mu/kappa receptors bound [3H] diprenorphine with high affinities, indicating that these chimeras retain opioid receptor conformation. Binding affinities of three peptide ligands (dynorphin A, alpha-neo-endorphin, and dynorphin B) and three nonpeptide ligands (norbinaltorphimine, U50,488H, and U69,593) for chimeras were determined and compared to those for mu and kappa opioid receptors. The second extracellular loop and the adjoining C-terminal portion of the fourth transmembrane helix were essential for the high affinity binding of dynorphin A, alpha-neo-endorphin, and dynorphin B to the kappa receptor. The third extracellular loop and the sixth and seventh transmembrane helices played an important role in determining the selectivity of nor-binaltorphimine for the kappa over the mu receptor. U50,488H and U69,593 appeared to require the whole kappa receptor except the second extracellular loop to attain high affinity binding. Thus, the kappa opioid receptor has differential binding domains for peptide and non-peptide ligands.

Intrasplenic metastasis of ovarian malignancies are uncommon. From 1986 to 1992 only 12 such cases were diagnosed, being 1.2% of the total number of patients with ovarian malignancies treated in tumor hospital during the same period. Of the 12 metastasis, 6 were poorly differentiated. Diagnosis was made through imaging techniques or surgery. The median interval from the diagnosis of the primary disease to intrasplenic involvement was 28 months. The metastatic lesions, presented themselves as single or multiple nodes within the cortex of the spleen. Clinical features were not remarkable. It was thought that the occurrence of intrasplenic involvement might be due to greater tumor bulk, poor differentiation and decreased host immuno-competence caused by chemotherapy. Most of the intrasplenic metastasis was found after liver metastasis or simultaneously with the latter. The main route of the tumor spread might be via blood vessels. Sole intrasplenic metastasis can be treated by surgery with a temporarily good result. Poor prognosis and high mortality characterize intrasplenic metastasis. The median survival was only 7 months.

For analysing the injurious mechanism of Trichosanthin (TCS) on trophoblast cells, cytotrophoblast cells were separated from placental villi in human early pregnancy and cultured on millipore filters coated with collagen in dual-environmental culture chambers. After 7-10 days culture the cells grew into confluent monolayer. A lot of multinucleated giant cells (synthetial like) could be found under light microscopy. Cytotrophoblast cells and the giant one were characterized as epithelial type by indirect immuno-fluorescent staining with anti-keratin. Because the procedure of separation of trophoblast cells is laborious and the choriocarcinoma cells (JAR) were as sensitive as trophoblast cells to trichosanthin, so the choriocarcinoma cells were used instead of the trophoblast cells in the later experiments. The internalization and distribution of TCS conjugated to 15 nm (in diameter) gold particles were examined. Electron microscopy showed that the TCS-gold particles were bound to the cell membrane and entered via coated pit and then internalized into coated vesicles (endosomes) within 30-60 minutes after treatment. Nevertheless, a number of free TCS-gold particles entered into the cell membrane nonspecifically. In the series of 60-120 minutes treatment, the TCS-gold particles presented in the multivesicular bodies. It is worthy to emphasize that the TCS-gold particles entered the cytosol from the endosomes and distributed nearby the rough endoplasmic reticulum (RER) and ribosomes within 120-180 minutes. In the meanwhile, the cells showed pathological changes markedly. With the same method, human liver carcinoma cells (BE 7402), kidney cells of African green monkey (CV-1) and rat embryonic liver epithelial cells (LW 13) were treated by TCS-gold particles as control. However, no particles had been found on the cell surface or in the cytosol within 60-120 minutes (see Table 1). In addition, trophoblast cells were treated by BSA-gold particles and transferrin-gold particles separately there still no particles could be found. (See Table 2). It is more interesting that TCS-Hepama-1-gold particles internalized into the choriocarcinoma cells with the same manner as TCS-gold entered, and it could not be affected by pretreatment with Hepama-1 an hour. But, TCS-Hepama-1-gold particles bound to the microvilli of human liver carcinoma cells, and it can be inhibited competitively by pretreatment with Hepama-1 for an hour (see Table 3). These results indicated consistently that trichosanthin possesses a high affinity to the membrane of trophoblast cells and choriocarcinoma cells, and the internalization of this plant toxic protein into its target cells characterized by receptor-mediated endocytosis.(ABSTRACT TRUNCATED AT 400 WORDS)

We previously showed that under defined conditions beta-[3H]funaltrexamine (beta-[3H]FNA) covalently labeled mu-opioid receptors with high specificity in bovine striatal membranes. beta-[3H]FNA-labeled mu-opioid receptors migrated as a broad band with a molecular mass range of 68-97 kDa. It is controversial whether beta-FNA binds irreversibly to mu-opioid receptors in other species. In this study, we demonstrated that beta-[3H]FNA also labeled mu-opioid receptors with high specificity in brain membranes of the guinea pig, rat, and mouse. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography revealed that in each species beta-[3H]FNA specifically bound to a protein in which labeling was greatly reduced by naloxone. These labeled receptors had broad molecular mass ranges, and the molecular masses were different among these species, in the order of cowguinea pigratmouse. Membranes were subjected to solubilization with 2% Triton X-100 and wheat germ lectin (WGL) affinity chromatography. N-Acetylglucosamine eluted a peak of radioactivity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography showed that in all four species the mu receptor was the only protein labeled with beta-[3H]FNA in the WGL eluate. The molecular masses of labeled mu-opioid receptors were 70-88 kDa (median, 77 kDa) for the cow, 66-80 kDa (median, 72 kDa) for the guinea pig, 60-75 kDa (median, 67 kDa) for the rat, and 60-72 kDa (median, 66 kDa) for the mouse. In addition, we investigated the nature of the carbohydrate moieties linked to the receptor protein and whether the species variation in the molecular mass was due to variable degrees of glycosylation. The bovine WGL eluate was treated with various glycosidases. Neuraminidase treatment decreased the receptor molecular mass by 6-7 kDa, whereas alpha-mannosidase had no effect. Removal of N-linked carbohydrates at asparagine residues by peptide-N4-[N-acetyl-beta-glucosaminyl]asparagine amidase (N-Glycanase) resulted in a much sharper specifically labelled protein band of 43 kDa. These results indicate that mu-opioid receptors are heavily glycosylated and the major carbohydrate moieties are of the complex type, N-linked to asparagine. After the WGL eluates for the four species were treated with N-Glycanase, the labeled receptors became much sharper bands with very similar molecular masses, i.e., 43 kDa for the cow and guinea pig, 39 kDa for the rat, and and 40 kDa for the mouse.(ABSTRACT TRUNCATED AT 400 WORDS)