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We report on the utility of forward-planned, 3-dimensional (3D), multiple-segment tangential fields for radiation treatment of patients with breast cancer. The technique accurately targets breast tissue and the tumor bed and reduces dose inhomogeneity in the target. By decreasing excess dose to the skin and lung, a concomitant boost to the tumor bed can be delivered during the initial treatment, thereby decreasing the overall treatment time by one week. More than 120 breast cancer patients have been treated with this breast conservation technique in our clinic. For each patient, a 3D treatment plan based upon breast and tumor bed volumes delineated on computed tomography (CT) was developed. Segmented tangent fields were iteratively created to reduce "hot spots" produced by traditional tangents. The tumor bed received a concomitant boost with additional conformal photon beams. The final tumor bed boost was delivered either with conformal photon beams or conventional electron beams. All patients received 45 Gy to the breast target, plus an additional 5 Gy to the surgical excision site, bringing the total dose to 50 Gy to the boost target volume in 25 fractions. The final boost to the excision site brought the total target dose to 60 Gy. With minimum follow-up of 4 months and median follow-up of 11 months, all patients have excellent cosmetic results. There has been minimal breast edema and minimal skin changes. There have been no local relapses to date. Forward planning of multi-segment fields is facilitated with 3D planning and multileaf collimation. The treatment technique offers improvement in target dose homogeneity and the ability to confidently concomitantly boost the excision site. The technique also offers the advantage for physics and therapy staff to develop familiarity with multiple segment fields, as a precursor to intensity-modulated radiation therapy (IMRT) techniques.

The binding of tumor necrosis factor alpha (TNF-alpha) to the type-1 TNF receptor (TNFRc1) plays an important role in inflammation. Despite the clinical success of biologics (antibodies, soluble receptors) for treating TNF-based autoimmune conditions, no potent small molecule antagonists have been developed. Our screening of chemical libraries revealed that N-alkyl 5-arylidene-2-thioxo-1,3-thiazolidin-4-ones were antagonists of this protein-protein interaction. After chemical optimization, we discovered IW927, which potently disrupted the binding of TNF-alpha to TNFRc1 (IC(50) = 50 nM) and also blocked TNF-stimulated phosphorylation of Ikappa-B in Ramos cells (IC(50) = 600 nM). This compound did not bind detectably to the related cytokine receptors TNFRc2 or CD40, and did not display any cytotoxicity at concentrations as high as 100 microM. Detailed evaluation of this and related molecules revealed that compounds in this class are "photochemically enhanced" inhibitors, in that they bind reversibly to the TNFRc1 with weak affinity (ca. 40-100 microM) and then covalently modify the receptor via a photochemical reaction. We obtained a crystal structure of IV703 (a close analog of IW927) bound to the TNFRc1. This structure clearly revealed that one of the aromatic rings of the inhibitor was covalently linked to the receptor through the main-chain nitrogen of Ala-62, a residue that has already been implicated in the binding of TNF-alpha to the TNFRc1. When combined with the fact that our inhibitors are reversible binders in light-excluded conditions, the results of the crystallography provide the basis for the rational design of nonphotoreactive inhibitors of the TNF-alpha-TNFRc1 interaction.

Heterologous proteins expressed on the surface of cells may be useful for eliciting therapeutic responses and engineering new extracellular properties. We examined factors that control the membrane targeting of alpha-fetoprotein (AFP) and a single-chain antibody (scFv). Chimeric proteins were targeted to the plasma membrane by employing the transmembrane domain (TM) and cytosolic tail of murine CD8O (B7-1), the TM of the human platelet-derived growth factor receptor (PDGFR), the glycosylphosphatidylinositol anchor encoded by the C-terminal extension of decay-accelerating factor (DAF), and the TM of the H1 subunit of the human asialoglycoprotein receptor (ASGPR). AFP chimeric proteins containing the B7, DAF, ASGPR, or PDGFR targeting domains displayed half-lives of 12.2, 3.8, 2.4, and 1.6 h, respectively. The newly synthesized B7 chimera was rapidly transported and remained on the cell surface. Glycosylphosphatidylinositol-anchored chimeras reached the surface more slowly and significant amounts were released into the culture medium. PDGFR TM chimeras were rapidly degraded, whereas ASGPR chimeras were retained in the endoplasmic reticulum (ER). The surface expression of both AFP and scFv chimeric proteins followed the order (highest to lowest) of B7 > DAF >> PDGFR. Introduction of a dimerization domain (hinge-CH(2)-CH(3) region of human IgG1) between scFv and TM dramatically reduced cleavage of the chimeric protein, increased surface expression, and produced biologically active scFv. Our results indicate that transgenes designed for the expression of active scFv on cells should incorporate a TM that does not undergo endocytosis, include an intact cytoplasmic domain, and possess a spacer to reduce cleavage and retain biological activity., (Copyright 2001 John Wiley & Sons, Inc.)

Although Candida rugosa utilizes a nonuniversal serine codon (CUG) for leucine, it is possible to express lipase genes (LIP) in heterologous systems. After replacing the 19 CUG codons in LIP4 with serine codons by site-directed mutagenesis, a recombinant LIP4 was functionally overexpressed in Pichia pastoris in this study. This recombinant glycosylated lipase was secreted into the culture medium with a high purity of 100 mg/liter in a culture broth. Purified recombinant LIP4 had a molecular mass of 60 kDa, showing a range similar to that of lipase in a commercial preparation. Since LIP4 has only a glycosylation site at position Asn-351, this position may also be the major glycosylation site in C. rugosa lipases. Although the thermal stability of recombinant LIP4 significantly increased from 52 to 58 degrees C after glycosylation, there were no significant differences in the catalytic properties of recombinant glycosylated lipase from P. pastoris and the unglycosylated one from Escherichia coil. These two recombinant LIP4s showed higher esterase activities toward long-chain ester (C16 and C18) and exhibited higher lipase activities toward unsaturated and long-chain lipids. In addition, LIP4 does not show interfacial activation as compared with LIP1 toward lipid substrates of tributyrin and triolein. These observations demonstrated that LIP4 shows distinguished catalytic activities with LIP1 in spite of their high sequence homology.

Purpose: To evaluate the miniature multileaf collimator (MMLC) as an alternative to traditional circular collimators for radiosurgery., Materials and Methods: 'Circular' fields were created with the Radionics MMLC (leaf width 3.53 mm at isocenter). Beam data, including tissue maximum ratios, output factors, penumbrae and isodose distributions of these fields were measured. These were compared to the Radionics circular collimators traditionally used for radiosurgery. The MMLC data were input to the XKnife Treatment Planning System. Treatment plans were completed and evaluated using both the MMLC 'circular' fields and the circular collimators., Results: MMLC fields using 3, 5, 7, 9, 11, and 13 leaves on each side of the Radionics MMLC were created to approximate circular fields. The TMRs are essentially identical to those of comparable-size circular collimators. Measured at isocenter at 5-cm depth for 6 MV, the 80-20% penumbra widths are comparable to circular collimators, but are increased by as much as 1 mm at the leaf intersections (steps) where scalloping occurs. Isodose distributions were matched to those of circular collimators with comparable 50% isodose widths. Treatment plans for the MMLC 'circular' fields with four arcs (totaling 360 degrees) are essentially identical to those of comparable circular collimators. Dose-volume histograms revealed clinically insignificant differences between the two in doses to the target, to the volume surrounding the target, and to adjacent critical normal tissues. There is very little discrepancy between the dose distribution calculated with the approximated MMLC fields and with those of simulated arcs with the actual MMLC fields., Conclusions: With the MMLC simulating circular fields, dose distributions may be obtained which are essentially identical to comparable-size circular collimators. The mechanical accuracy of the MMLC is as good as that of the circular collimators, and the leakage dose is less. The diameter of 'circular' fields is limited by the MMLC leaf width to 1 cm and greater in increments of 7 mm. Attention needs to be paid to mechanical collisions because the MMLC is bulkier than the circular collimators., (Copyright 2002 S. Karger AG, Basel)

Immunohistochemistry using a StAR peptide antiserum had previously revealed strong staining in rat and bovine adrenal medulla, suggesting the presence of a protein immunogenically related to StAR. Western blots of bovine medulla tissue homogenates showed the principal adrenal medullary immuno-reactive species to have a higher molecular weight (50 kDa) compared to StAR protein (30 kDa). Subcellular fractionation localised the 50 kDa species principally to the medulla cytosol. StAR peptide antiserum binding to both the 30 kDa and 50 kDa species could be specifically competed by the peptide antigen. These data suggest that the adrenal medullary immuno-reactive species and StAR protein are distinct entities, which share some features in common.

Previous studies using keratinocytes from epidermal growth factor receptor (EGFR)-deficient mice revealed that the EGFR is not required for papilloma formation initiated by a mutant rasHa gene, although the tumors that develop are very small (A. A. Dlugosz et aL, Cancer Res., 57: 3180-3188, 1997). The current study used a combination of bromodeoxyuridine pulse-chase, proliferating cell nuclear antigen distribution, and differentiation marker analysis to reveal the following: (a) the EGFR was required to maintain the proliferative population in the basal cell compartment of papillomas; (b) in the absence of EGFR, cycling tumor cells migrated into the suprabasal compartment and initiated the differentiation program prematurely; and (c) these changes were associated with cell cycle arrest. Further analysis of v-rasHa-transformed EGFR-deficient keratinocytes in vitro indicated that such cells migrated more on and attached less to extracellular matrix components. Together, these studies reveal that an essential function for the EGFR pathway in squamous tumors is to maintain a proliferative pool of basal cells and prevent premature terminal differentiation.

The Escherichia coli thioesterase I specifically catalyzes the deacylation of fatty acyl-CoA thioesters, especially those with long acyl groups (C(12)-C(18)). Single crystals of thioesterase I (E.C. 3. 1.2.2) from E. coli have been obtained using methoxypolyethylene glycol 5000 (PEG-MME 5K) as a precipitant at room temperature in 21 d. The crystals belong to the tetragonal space group P4(1)2(1)2 or its enantiomorph P4(3)2(1)2, with unit-cell parameters a = b = 50.85 (7), c = 171.5 (1) A. The crystals diffract to beyond 2.4 A resolution. There is one molecule of molecular weight 20.5 kDa in the asymmetric unit, with a solvent content of 55%.

Purpose: To describe a method of creating an intraoperative dosimetric representation of the real-time ultrasound-guided prostate implant., Materials and Methods: An intraoperative dosimetry system (Multi Media Systems [MMS]) captures transverse ultrasound images after peripheral needles have been implanted in the prostate. The prostate contour and needle positions are outlined on the system. The volume of the prostate with needles in place is calculated. As seeds are deposited in the actual implant, the positions of the seeds are marked on the intraoperative system. Following implantation of the peripheral needles, the resulting isodose lines are displayed. The interior needles are inserted into the prostate, and these positions are captured on the system. As seeds are deposited through these needles into the prostate, their positions are captured on the planning system. When the implant is complete, the final dose coverage and dose volume histogram can be visualized., Results: Ten consecutive patients underwent iodine 125 implants using real-time intraoperative isodose generation. The ratio of the preneedle prostate volume to postneedle prostate volume ranged from 0.89 to 1.0 (median 0.97). The calculated dose delivered to 90% of the prostate volume from the dose volume histogram (D90) ranged from 146.5 to 194 Gy (median 174.75 Gy). The percentage of the prostate covered by 240 Gy ranged from 14.6% to 59% (median 40.75%)., Conclusion: Dosimetric representation of the real-time ultrasound-guided prostate implant can be achieved and demonstrates the efficacy of this brachytherapy technique.

Activation of cytotoxic T cells without MHC restriction was attempted by expressing single-chain antibodies (scFv) against CD3 on the surface of tumor cells. A chimeric protein consisting of a scFv of mAb 145.2C11, the hinge-CH2-CH3 region of human IgG1, and the transmembrane and cytosolic domains of murine CD80 formed disulfide-linked dimers on the plasma membrane of cells and specifically bound lymphocytes. Anti-CD3 scFv dimers expressed on the cell surface induced CD25 (IL-2 receptor alpha-chain) expression and proliferation of splenocytes. CT26 tumor cells engineered to express surface scFv dimers (CT26/2C11) also induced potent lymphocyte cytotoxicity with or without addition of exogenous IL-2. Splenocytes activated by CT26/2C11 cells also killed wild-type CT26 cells, indicating that activated splenocytes could kill bystander tumor cells. Immunization of BALB/c mice with irradiated CT26/2C11 cells did not protect against a lethal challenge of CT26 cells, suggesting that systemic immunity was not induced. However, the growth of CT26 tumors containing 50% CT26/2C11 cells was significantly retarded compared with CT26 tumors, whereas CT26/2C11 tumors did not grow in syngeneic mice. These results suggest that expression of anti-CD3 scFv dimers on tumors may form the basis for a novel therapeutic strategy for tumors that exhibit defects in antigen processing or presentation. Gene Therapy (2000) 7, 339-347.

The effects of tetrandrine, a blocker of voltage-dependent Ca(2+) channels, on ionic currents were investigated in an endothelial cell line (HUV-EC-C) originally derived from human umbilical vein. In whole-cell configuration, tetrandrine (0.5-50 microM) reversibly decreased the amplitude of K(+) outward currents. The IC(50) value of tetrandrine-induced decrease in outward current was 5 microM. The K(+) outward current in response to depolarizing voltage pulses was also inhibited by iberiotoxin (200 nM), yet not by glibenclamide (10 microM) or apamin (200 nM). The reduced amplitude of outward current by tetrandrine can be reversed by the further addition of Evans' blue (30 microM) or niflumic acid (30 microM). Thus, the tetrandrine-sensitive component of outward current is believed to be Ca(2+)-activated K(+) current. Pretreatment with thapsigargin (1 microM) or sodium nitroprusside (10 microM) for 5 h did not prevent tetrandrine-mediated inhibition of outward current. In outside-out configuration, bath application of tetrandrine (5 microM) did not change the single-channel conductance but significantly reduced the opening probability of large-conductance Ca(2+)-activated K(+) (BK(Ca)) channels. The tetrandrine-mediated decrease in the channel activity was independent on internal Ca(2+) concentration. Tetrandrine (5 microM) can also shift the activation curve of BK(Ca) channels to more positive potentials by approximately 20 mV. The change in the kinetic behavior of BK(Ca) channels caused by tetrandrine is due to a decrease in mean open time and an increase in mean closed time. The present study provides substantial evidence that tetrandrine is capable of suppressing the activity of BK(Ca) channels in endothelial cells. The direct inhibition of these channels by tetrandrine should contribute to its effect on the functional activities of endothelial cells.

A gene involved in quinate metabolism was cloned from Xanthomonas campestris pv. juglandis strain C5. The gene, qumA, located on a 4. 2-kb KpnI-EcoRV fragment in plasmid pQM38, conferred quinate metabolic activity to X. c. pv. celebensis. Tn3-spice insertional analyses further located the qumA gene on a region of about 3.0 kb within pQM38. Nucleotide sequencing of this 3.0-kb fragment reveals that the coding region of qumA is 2373 bp, the deduced amino acid sequence of which closely resembles a pyrrolo-quinoline quinone-dependent quinate dehydrogenase of Acinetobacter calcoaceticus. A 0.7 kb SalI-PstI fragment internal to qumA was used as a probe to hybridize against total genomic DNA from 43 pathovars of X. campestris. The fragment hybridized only to total genomic DNA from the four pathovars of DNA homology group 6, X. c. pv. celebensis, X. c. pv. corylina, X. c. pv. juglandis and X. c. pv. pruni, and from X. c. pv. carotae, which belongs to DNA homology group 5. This 0.7 kb fragment was also used as a probe to hybridize BamHI-digested total genomic DNAs from the four pathovars of DNA homology group 6 and X. c. pv. carotae. The restriction fragment length polymorphism pattern of DNA homology group 6 was different from that of X. c. pv. carotae. The probe hybridized to a 5.7-kb BamHI fragment in all four pathovars of group 6 and to a 6.1-kb BamHI fragment in three of four pathovars. It hybridized only to a 9. 9-kb BamHI fragment in X. c. pv. carotae. Quinate metabolism has previously been reported as a phenotypic property specific to X. campestris DNA homology group 6. Accordingly, a combination of the quinate metabolism phenotypic test and Southern hybridization using a qumA-derived probe will be very useful in the identification of pathovars in DNA homology group 6.

Targeting of proteins to the plasma membrane of cells may be useful for vaccine development, tissue engineering, genetic research, bioseparations, and disease treatment. The ability of different transmembrane domains (TM) to direct a reporter protein (human alpha-feto protein, AFP) to the surface of mammalian cells was examined. High surface expression was achieved with chimeric proteins composed of AFP and the TM and cytosolic tail of murine B7-1 (AFP-B7) as well as with AFP containing a GPI-anchor from decay-accelerating factor (AFP-DAF). Lower surface expression of AFP was observed when the TM of human platelet-derived growth factor receptor or the human asialoglycoprotein receptor H1 subunit were employed. Introduction of the hinge-CH2-CH3 region of human IgG (gamma1 domain) between AFP and TM allowed efficient formation of disulfide-linked dimers. Surface expression of AFP-gamma1-B7 dimers was impaired compared to AFP-B7 whereas AFP-gamma1-DAF dimers were efficiently targeted to the surface. Accumulation of chimeric proteins on the cell surface did not correlate with the level of protein expression. This study demonstrates that high levels of monomeric and dimeric proteins can be targeted to the cell membrane of mammalian cells by proper selection of TM., (Copyright 1999 John Wiley & Sons, Inc.)

Isoeugenolol (1.0, 3.0, 5.0 mg/kg, i.v.) produced a dose-dependent bradycardia and a decrease in blood pressure in anesthetized Wistar rats. Isoeugenolol inhibited the tachycardia effects induced by (-)isoproterenol, but had no blocking effect on the arterial pressor responses induced by (-)phenylephrine. In isolated guinea pig tissues, isoeugenolol antagonized (-)isoproterenol-induced positive inotropic and chronotropic effects on the atria and tracheal relaxations in a concentration-dependent manner. The apparent pA2 values for isoeugenolol on right atria, left atria and trachea were 7.63+/-0.03, 7.89+/-0.12 and 6.12+/-0.05, respectively, indicating that isoeugenolol was a highly selective beta1-adrenoceptor blocker. On the other hand, isoeugenolol produced a mild direct cardiac depression at high concentration and was without intrinsic sympathomimetic activity (ISA). In isolated rat thoracic aorta, isoeugenolol relaxed more potently the contractions induced by (-)phenylephrine (10 microM) and 5-HT (10 microM) than those by high K+ (75 mM). In isolated guinea pig trachea, isoeugenolol attenuated the carbachol (1 microM)-con-tracted trachea more significantly than those contracted with high K+. Furthermore, the binding characteristics of isoeugenolol and various beta-adrenoceptor antagonists were evaluated in [3H]CGP-12177 binding to rat ventricle, lung and interscapular brown adipose tissue (IBAT) membranes. The -log IC50 values of isoeugenolol for predominate beta1-, beta2- and beta3-adrenergic receptor sites were 5.82+/-0.09, 4.74+/-0.05 and 4.73+/-0.12, respectively. In conclusion, isoeugenolol was found to be a highly selective beta1-adrenoceptor antagonist with tracheal and vascular smooth muscle relaxant activities, but was devoid of alpha-adrenoceptor-blocking action.

Escherichia coli has seldom been reported to cause pseudobacteremia. The investigation of an outbreak of amoxycillin-clavulanic acid-resistant E. coli pseudobacteremia is described. Seventeen cases occurred over a five-day period. The source of the E. coli was traced to the blood culture specimen of a patient (index patient) with genuine bacteremia as a result of urinary tract infection. The other 16 case-patients had pseudobacteremia which was found to be the result of cross-contamination during subculture of blood specimens. The E. coli strain was carried over from the culture bottle of the index patient, through the contaminated gloved hands of a technician to the culture bottles of the other 16 cases. Although the pseudobacteremia occurred over a five-day period, they all resulted from cross-contamination during blood culture processing within one day. An early outbreak investigation was prompted by the unusual finding of amoxycillin-clavulanic acid resistance in the case E. coli isolates in a short period. The relatedness of the E. coli strains from the 17 cases was confirmed by arbitrary-primed polymerase chain reaction. Clinicians should be alerted to the possibility of a blood E. coli isolate being a contaminant despite its predominant role as a true pathogen.

In this study, the aorta vasorelaxant, coronary calcitonin gene-related peptide (CGRP) releasing, and atrial contractility effects of glyceryl nonivamide (GLNVA) were investigated in guinea pigs. In the isolated thoracic aorta, although GLNVA (0.01-50 microM) concentration dependently induced endothelium-independent relaxations and relaxed phenylephrine-(1.0 microM) induced contractions, it failed to relax 80 mM KCI-induced contractions. The GLNVA (1.0 microM) relaxation response in the aorta was significantly inhibited by tetraethylammonium (2.5-10 mM) or ouabain (5.0 microM) and was attenuated by increased extracellular potassium gradient (10-30 mM). Glibenclamide (0.01-10 microM) dose dependently antagonized the GLNVA relaxant effect. In the isolated perfused guinea pig heart, GLNVA (0.1-10 microM) increased CGRP-like immunoreactivity outflow from coronary circulation in a concentration-dependent manner. In the isolated right and left guinea pig atria, GLNVA (0.01-10 microM) produced concentration-dependent positive inotropic and chronotropic effects, but these effects were inhibited by pretreatments with ruthenium red (1.0 microM), capsazepine (10 microM), human calcitonin-gene-related peptide (CGRP(8-37)) (1.0 microM) and sensory neuron denervation, respectively. Based on these findings, we suggest that CGRP may be released by GLNVA from cardiovascular sensory neuron, and it then activates CGRP receptors on the coronary artery and atrium. The GLNVA-induced vasorelaxant effect in the vascular smooth muscle of the aorta is due to CGRP release associated K+ channel opening, and this effect eliminates capsaicin-derived excitability-associated K+ channel blocking activities.

Effects of glyceryl nonivamide (GLNVA), a nonpungent analogue of capsaicin, on spinal substance P release, blood pressure and renal function were investigated in rats. GLNVA was administered intrathecally or intracerebroventricularly, and renal clearance function was measured. The addition of GLNVA at concentrations of 2, 20 and 200 mumol/l during in vivo spinal superfusion resulted in concentration-dependent increases in the release of substance P. Capsaicin pretreatment (50 mg/kg/day, s.c. for 2 weeks) blunted the GLNVA-induced substance P release, suggesting a stimulation of GLNVA on the spinal terminals of capsaicin-sensitive primary afferent nerves to release substance P. Intracerebroventricular administration of GLNVA at concentrations of 20, 40 and 80 nmol/l evoked a concentration-dependent depressor response. Despite the hypotensive effect, GLNVA caused significant increases in glomerular filtration rate, urine flow, absolute and fractional excretion of sodium and potassium, osmolar clearance and free water reabsorption rate. These data suggest that central administration of GLNVA stimulates spinal substance P release, reduces blood pressure but enhances renal excretion of water and electrolytes due to an increased filtered load and a decreased tubular reabsorption.

1. The beta-adrenoceptor blocking and calcitonin gene-related peptide (CGRP)-releasing properties of capsinolol (N-[4-(2-hydroxy-3 (isopropylamino) propoxy)-3-methoxybenzyl]-nonanamide), derived from nonivamide, were investigated under in vivo and in vitro conditions. 2. Capsinolol (0.1, 0.5, 1.0 mg kg-1, i.v.), as well as (+/-)-propranolol, produced a dose-dependent bradycardia response and a temporary pressor action in urethane-anaesthetized normotensive Wistar rats. These cardiovascular effects were different from the vagus reflex and parasympathetic efferent effects shown by capsaicin (0.1 mg kg-1, i.v.) in the rat. 3. Capsinolol (1.0 mg kg-1) inhibited the tachycardia effects induced by (-)-isoprenaline, but had no blocking effect on the arterial pressor responses induced by (-)-phenylephrine. The findings suggest that capsinolol possesses beta-adrenoceptor blocking activity, but it has no alpha-adrenoceptor blocking activity. 4. In guinea-pig isolated tissues, capsinolol (10(-8) to 10(-6) M) antagonized (-)-isoprenaline-induced positive chronotropic and inotropic effects of the atria and tracheal relaxation responses in a concentration-dependent manner. The parallel shift to the right of the concentration-response curve of (-)-isoprenaline suggests capsinolol is a beta-adrenoceptor competitive antagonist. 5. Capsinolol (10(-5) to 10(-4) M) exhibited a positive cardiotonic effect that was not inhibited by (+/-)-propranolol and reserpine, but was inhibited by capsazepine (10(-6) M) and CGRP8-37 (10(-6) M). This effect was independent of intrinsic sympathomimetic effects. 6. An immunoassay of released CGRP from guinea-pig isolated perfused heart indicated that capsinolol increases the release of CGRP and thus produces positive cardiotonic effects. 7. In conclusion, capsinolol is a non-selective beta-adrenoceptor antagonist with capsaicin-like cardiotonic properties unrelated to traditional intrinsic sympathomimetic effects. It is suggested that capsinolol causes CGRP release from cardiac sensory neurones via a non-adrenergic mechanism and then activates CGRP receptors on cardiac muscle.

The electrophysiological effects of capsaicin in rat atrial myocytes were examined. Measurement of contractile force was done in rat left atria. Whole-cell patch-clamp technique was primarily used to study the change in membrane potential and ionic currents. Capsaicin produced an initial rise and a sustained increase in contractile force in rat left atria. Capsaicin (10 μM) caused a significant prolongation of atrial action potential. In voltage-clamp experiments, capsaicin (1-100 μM) caused the reversible reduction in the amplitude of transient outward (I(TO)) and late outward (I(L)) K(+) currents in concentration- and voltage-dependent manners. The time course for inactivation of I(TO) was changed to the biexponential process after the application of capsaicin. Capsaicin failed to cause any significant shift in quasi-steady-state inactivation curve of I(TO). The EC(50) values for the inhibitory effects of capsaicin on I(TO) and I(L) were 5 and 20 μM, respectively. Capsaicin also suppressed the amplitude of acetylcholine- or adenosine-induced K(+) current, i.e., I(K(ACh,Ado)). The EC(50) value for capsaicin-mediated inhibition of I(K(ACh,Ado)) is 50 μM. The present findings suggest that in isolated rat atria, during capsaicin exposure, the capsaicin-mediated inhibition of these K(+) channels is one of the ionic mechanisms underlying the positive inotropic and chronotropic actions.

The ultra-short-acting and vasorelaxant beta 1-adrenoceptor blocking activities of vasomolol, a guaiacoxypropanolamine derivative of vanillic acid ethyl ester, were studied. Vasomolol (0.5, 1.0, 3.0 mg/kg intravenously, i.v.) produced a dose-dependent bradycardia response and demonstrated particularly a hypotensive action with an ultra-short-acting property in pentobarbital-anesthetized normotensive rats. Vasomolol's steady state of beta-blockade was attained < or = 10 min after initial infusion, and a rapid recovery from blockade occurred after discontinuation of the infusion, although intravenous infusion of vasomolol (300 micrograms/kg/min) could not inhibit pressor responses induced by (-)phenylephrine (10 micrograms/kg i.v.). In isolated rat thoracic aorta, vasomolol (1-10 microM) inhibited vascular smooth muscle contractions induced by both (-)phenylephrine (10(-5) M) and high K+ (75 mM) concentration dependently. This inhibitory effect of vasomolol was more sensitive on K(+)-induced than on (-)phenylephrine-induced contractions, suggesting that the block of Ca2+ influx may involve the major mechanism of vasorelaxation. In isolated guinea pig tissues, vasomolol (0.01-10 microM) antagonized the (-)isoproterenol (ISO)-induced positive inotropic and chronotropic effects of the atria and tracheal relaxation responses in a concentration-dependent manner. The parallel shift to the right of the concentration-response curve of (-)ISO suggested that vasomolol was a beta-adrenoceptor competitive antagonist. The effect of vasomolol was more potent on atria than on tracheal tissues, indicating that it possesses beta 1-adrenoceptor selectivity. In addition, vasomolol did not show intrinsic sympathomimetic activity (ISA). Moreover, the binding characteristics of vasomolol were evaluated in [3H]dihydroalprenolol ([3H]DHA) binding to porcine ventricular membranes. Vasomolol was an ultra-short-acting and highly selective beta 1-adrenoceptor antagonist with vasorelaxant activity and is devoid of ISA.

Objectives: To study the safety, tolerability of and immune response to an inactivated hepatitis A vaccine in healthy Chinese adults., Design: Seronegative subjects were randomized to receive formalin-inactivated, alum-adjuvanted hepatitis A vaccine (25 units, MSD) at 0, 24 weeks (group I) or 0, 2, 24 weeks (group II)., Subjects: Healthy Chinese adults aged 18 years or above were recruited. All subjects were staff of a regional hospital and primary care clinic in Hong Kong., Main Outcome Measures: Side-effects, tolerance, seroconversion rate and geometric mean titre (GMT) of the two groups were noted and compared., Results: Thirty-two and 28 people were recruited into group I and group II, respectively. Antibody against hepatitis A developed in 100% of the recipients 1 month after the last dose of either regimen, with a GMT of around 450 IU L-1. High antibody levels were maintained at the end of one year. There was no statistically significant difference in the seroconversion rates and GMTs between the two regimens of vaccination. All subjects in group I seroconverted at 24 weeks after one single dose of the vaccine. No adverse effects were reported except for complaints of some mild local and constitutional symptoms., Conclusions: The inactivated hepatitis A vaccine is highly immunogenic, safe and well-tolerated.

Objective: To study the clinical and epidemiological characteristics of the first 100 reported AIDS cases in Hong Kong., Patients and Methods: The case records of the first 100 AIDS cases reported to the Department of Health were retrieved. An analysis was made on their demographic data, clinical and immunologic profile, treatment received as well as mortality., Results: The AIDS patients were reported over a ten-year period, from February 1985 to March 1994. The majority of them were male, Chinese and between the age of 20 and 49. More than 80% acquired HIV via sexual contact. The most common primary AIDS- defining illness was Pneumocystis carinii pneumonia (PCP), which occurred in 46 patients. Eight episodes of Penicillium marneffei infection have been recorded. Fifty-nine patients have received antiretroviral therapy whereas PCP prophylaxis was given to 47 patients. The mean CD4 count at HIV and AIDS diagnosis was 394/microliters and 121 /microliters respectively. Sixty-seven patients were known to have died at the time of the analysis. PCP was the most common cause of death, accounting for over one-fifth of the cases. The median survival after AIDS diagnosis was less than 2 months for cases reported in 1985-1987 and over 15 months for those of 1990-1991., Conclusion: Pneumocystis carinii pneumonia was the most common initial AIDS-defining disease and cause of death in Hong Kong. Survival of the patients has gradually improved over the past few years.

Sharing of contaminated injection equipment accounts for the rapid spread of the human immunodeficiency virus (HIV) among injecting drug users (IDUs). The profile of drug addiction practice among inmates of the Shek Kwu Chau Drug Rehabilitation Centre in Hong Kong was studied. Registers on all the new admissions to the Centre during a two-year period between 1990 and 1992 were reviewed. Of the 3129 drug users studied, 68.7% were aged between 21 and 40; 84.8% were IDUs and heroin was the commonest drug of addition. Nearly 70% of the IDUs had never shared injection equipment with others. There were significantly more young addicts (< or = 30 years old) who had shared needles compared with the older ones (31.2% vs 26.8%, P < 0.05). Those with addiction time > 6 months were more likely to have shared needles than the new ones. Only 19% of the drug users accepted HIV testing at their first admission. Factors speculated for the low needle-sharing rates among IDUs in Hong Kong and the low HIV prevalence in the IDU population are discussed. It is of utmost importance to monitor continuously such a high-risk behaviour pattern so as to design appropriate intervention strategies to stop the transmission of HIV.

The application of capsaicin (1 microM) produced a minor relaxant effect in endothelium-denuded rat aortae. However, capsaicin caused a greater relaxation of blood vessels precontracted with high K+ or phenylephrine. The effects of capsaicin on the ionic currents were also examined in A7r5 vascular smooth muscle cells. The tight-seal whole-cell voltage clamp technique was used. Capsaicin inhibited the Ba2+ inward current (IBa) through the voltage-dependent L-type Ca2+ channel in a concentration-dependent fashion, whereas calcitonin gene-related peptide and phenylephrine produced a minor increase in IBa. Capsaicin did not alter the overall shape of current-voltage relationship of IBa. However, capsaicin (3 microM) shifted the quasi-steady-state inactivation curve of IBa to more negative membrane potential by about 5 mV. These effects of capsaicin on IBa were reversible. In addition, capsaicin had inhibitory effects on voltage dependent K+ currents. These results suggest that inhibition of the voltage-dependent L-type Ca2+ channel is involved in the capsaicin-induced relaxation of the vascular smooth muscle, whereas capsaicin-induced inhibition of voltage-dependent K+ channels might produce an increase in cell excitability.

The effectiveness of a 2.5 micrograms dose of the hepatitis B vaccine (B-Hepavac II) was compared with that of 5 micrograms in 587 low-risk neonates and 777 preschool children of age 3-8 years. The vaccines were administered at months 0, 1 and 3, with postvaccination serology tested at months 4 and 12. The seroconversion rates of the 2.5 microgram recipients (newborn: 93.5%; preschool children: 97.4%) are comparable with the 5 micrograms group (newborn: 95.7%; preschool children: 98.7%). The seroconversion rates of the newborns are, however, significantly lower in the 2.5 micrograms group if positive response is taken as a titre > 10 IU l-1, instead of > 0 IU l-1. The older children, on the other hand, achieved a higher seroconversion rate and geometric mean titre (GMT) when compared with the newborns. irrespective of the dose received.

Background: To determine the spectrum of opportunistic infections in patients with human immunodeficiency virus (HIV-1) infection in Hong Kong., Methods: A retrospective study of 214 HIV-1-infected patients, seen between December 1984 and December 1993 in a specialist clinic for HIV/AIDS:, Results: A majority (94%) of the patients in the cohort were male; 84% had acquired HIV via sexual contacts. Two-thirds were ethnic Chinese. Ninety-two (43%) had developed AIDS, and 54(25%) had presented with other non AIDS-defining opportunistic infections during the study period. The primary AIDS defining illnesses of 80 patients were infections: Pneumocystis carinii pneumonia (50%), extrapulmonary tuberculosis (10%) and cytomegalovirus (CMV) disease (8%). Opportunistic infections among Chinese and non-Chinese were similar in spectrum, though higher frequencies of infection with CMV, Mycobacterium avium intracellulare and tuberculosis were seen among Chinese, whereas the opposite was true for Pneumocystis carinii pneumonia, toxoplasmosis and cryptosporidiosis. Disseminated Penicillium marneffei infection was another significant disease for HIV-positive patients. Common non AIDS-defining opportunistic infections included herpes zoster, oral candidiasis, herpes simplex infection and genital/anal wart. The median CD4 count at HIV diagnosis for AIDS patients was much lower than non-AIDS patients (147 vs 546/ul). Survival of deceased AIDS patients was poor, with a median of only five months. Survival has however, apparently, improved over the recent years., Conclusions: In Hong Kong, Pneumocystis carinii pneumonia remained the most common primary AIDS event, while Penicillium marneffei was emerging as another significant cause of major infection. Herpes zoster and oral candidiasis were the two most frequently encountered minor opportunistic infections.

We induced lipid peroxidation in rat heart mitochondria with FeSO4 and compared the inhibitory effect of various flavan-3-ol tannins on it. These tannins were purified from Chinese tea (Camellia sinensis). Oxygen consumption and malondialdehyde formation were used to quantitate the amount of lipid peroxidation. The free radical scavenger activity of tannins was then measured with a diphenyl-p-picrylhydrazyl method. These tannins significantly inhibited lipid peroxidation at micromolar concentration. Their potencies were higher than that of Trolox, a water soluble analogue of vitamin E. Since epicatechin-3-O-gallate, epigallocatechin-3-O-gallate and gallocatechin-3-O-gallate were more potent than other flavan-3-ol tannins in these assays, we considered that a galloyl group in 3-O-position increased the scavenger activity of flavan-3-ol tannins as well as their potency in inhibiting lipid peroxidation.

Purpose: The acute morbidity of doses of 64.8-75.6 Gy and preliminary observations of late complications and tumor response using 3-dimensional conformal radiation therapy in carcinoma of the prostate are assessed., Methods and Materials: 123 patients (Stage A2-12, B1-17, B2-43, C-51) were irradiated to the prostate and seminal vesicles using a 3-dimensional conformal radiation therapy technique. The median follow-up time was 15.2 months. The minimum tumor dose was 64.8-66.6 Gy in 49 patients, 70.2 Gy in 46, and 75.6 Gy in 28. Toxicity was scored according to the Radiation Therapy Oncology Group morbidity grading system., Results: This technique of 3-dimensional conformal radiation therapy was well-tolerated with minimal acute morbidity. Only 32% of patients had grade 2 or 3 acute morbidity requiring short-term medication for relief of urinary symptoms or diarrhea. Only one patient (0.8%) has so far developed a severe (grade 4) late complication. Serum prostate specific antigen concentrations normalized in 67% of patients (64/96) within 1-14 months (median 4.5 months) after treatment and were progressively decreasing at last measurement in an additional 22% (21/96). Abnormal rising prostate specific antigen levels were observed in 15 patients, 11 of whom have already developed other evidence of relapsing disease., Conclusion: Acute toxicity for the doses tested with this 3-dimensional conformal radiation therapy technique is reduced compared to traditional treatment techniques, and the initial tumor response as assessed by prostate specific antigen measurement is highly encouraging with prostate specific antigen levels returning to normal in the majority of patients. Based on these results, a further increase of the dose to 81 Gy has been implemented in accordance with the schema of an ongoing Phase I dose-escalation study.

Backscattering of radiation incident on a solid medium has been treated with some success as a reflection phenomenon. The concept of albedo has been used in the literature to deal with this problem. A Monte Carlo program has been used in this study to simulate the generation of bremsstrahlung beams from medical linear accelerators. The accuracy of the calculated spectra have been checked indirectly by calculating TPRs and comparing them with measured values. The dose albedos for 60Co and 137Cs beams normally incident on concrete have been calculated and compared with experimentally determined values in the literature. The generated energy spectra are then used to calculate the albedo factors for 4-, 10-, and 18-MV bremsstrahlung x-ray beams normally incident on concrete, iron, and lead. The number albedo decreases with increase in the atomic number of the reflecting medium. However, for high-energy photons, the dose albedo increases with increase in the atomic number of the medium. This is an important factor that needs to be considered in the design of shielding. The calculated energy spectra can be used in other problems and the albedo factors can be applied in practical shielding design for medical linear accelerators.

We have used a mouse model to study spinal cord injury following single doses (12 to 75 Gy) of radiation. The spinal cord (T9,10-L4,5) of C3Hf/Sed//Kam mice was irradiated and response graded using four levels of neurological change. Findings were: (a) the doses required to paralyze 50% of animals (ED50) were 19.79, 20.77, and 21.85 Gy for mild, partial, and complete paralysis, respectively, as measured 200-360 days after radiation. (b) Most damage was progressive but it was not necessarily so; after doses up to 28 Gy recovery was occasionally seen. (c) Latency depended on the dose and the level of damage. Following doses around the ED50, paralysis occurred between 180 to 300 days. (d) There were significant fluctuations in the dose-latency relationship at doses less than 35 Gy. Latency may be not a reliable endpoint to compare biological effects of radiation in this dose range. (e) The radiosensitivity of mouse spinal cord was similar to that reported for rats. (f) Histologically, demyelination was the dominant lesion in the paralyzed animals. We conclude that the mouse is a good animal model to study radiation damage to the spinal cord, at least when a 2.2 cm length is irradiated.

In many parts of human body, air-filled spaces are components of normal anatomical structures, such as nasopharynx and paranasal sinuses, and the inclusion of these spaces in radiation treatment fields may create inhomogeneous dose distribution, due to the loss of scattering electron equilibrium, the lack of backscatter and the alteration in primary beam attenuation. Accurate determination or measurement of dose in the region of irregular air spaces has been experimentally difficult. However, tumors growing on the surface of the air cavities do cause serious concern by conscientious radiation therapists and physicist about the possibility of underdose. Using the models of simulated air cavities, combined air cavity and air channel, and taking measurement with a small window parallel plate ionization chamber in solid phantoms, it is found that the air spaces in the human body will cause underdose or overdose at the exit air space surface for 10MV X-ray and Co-60 r-ray. Surface dose were evaluated by a gain/loss factors (G/L) which was defined as the ratio of exit air space surface dose and the dose measured in a same but homogeneous phantom. The G/L factors are greater than 1.0 (overdose) where the air spaces are much smaller than radiation field sizes and are less than 1.0 (underdose) in extreme situations where the air spaces are larger than radiation field sizes and where the scattered volume of cavity wall in the treatment fields in also small. In addition, these effects of overdose and underdose are enhanced by increasing air space thickness and higher photon energy. We suggest that the radiation therapists and clinical physicists be cautious in applying these useful guidelines for treatment planning.

The Superheated Drop Detector (SDD) is a new kind of neutron detector based on the same principle as that of the bubble chamber, except the superheated material is in drop form suspended by a gel or polymer as first described by Apfel (U.S. Patent 4,143,274). Previously, we have developed a theoretical model for the prediction of the threshold neutron energy to nucleate bubbles in our superheated materials and a model for the calculation of the energy dependent response function of SDD. The results of the theoretical calculations agree with the experimental measurements reasonably well. We found the measured response curve of one of our SDDs follows the same trend as the ideal ICRP dose equivalent response curve within 40% for neutrons of energy above 100 keV and within a factor of 10 below 100 keV. Therefore, it is possible to make a rem-response Superheated Drop Detector. We have further calibrated our SDD with 252Cf neutron sources (both bare and D2O moderated). The results are in reasonable agreement with our theoretical predictions, which proves that our model is valid not only for monoenergetic neutrons but also for neutrons from a radioactive source. The principles of operation of our detector, which can be used in neutron dosimetry, area monitoring, and neutron spectrometry, are discussed.

For decades, the high dose rate sources of Co-60 and Ir-192 have been used in remote controlled afterloading brachytherapy. A small volume ion chamber was used to measure the dose rate in varying distances from the sources. The entire dose distribution around the source was tabulated and the data were used to calculate the treatment time for individualized patient setting. A solid diode detector was used to plot the relative distance dose and the symmetrical dose distribution of the sources. Thermoluminescence dosimetry (TLD) was used for spot-checking of the dose. The dose distribution in the clinically useful range of 2-5 cm was carefully estimated. When normalized at 2.0 cm from the source, the dose rate at 5.0 cm was 17.1% for the Co-60 source and 18.4% for the Ir-192 source. It is clearly demonstrated that Ir-192 offers no dosimetric advantages, which is contrary to what used to be believed by some clinicians. The parameters of the computer treatment planning system have to be corrected and then adopted into clinical application.