Phylogenetic analysis of a coxsackievirus A24 variant: the most recent worldwide pandemic was caused by progenies of a virus prevalent around 1981.

Nucleotide substitutions in the viral-encoded proteinase 3C (3Cpro) region (549 nucleotides) of the RNA genome of a coxsackievirus A24 variant (CA24v), one of the agents causing acute hemorrhagic conjunctivitis (AHC), were studied using 32 isolates collected from the Eastern hemisphere in 1970-1989. Based on regression analysis of nucleotide differences among isolates, the nucleotide substitution rate of CA24v 3Cpro was estimated to be 3.7 x 10(-3)/nucleotide/year. A phylogenetic tree constructed by the modified unweighted pair group method using arithmetic averages (UPGMA) indicated that CA24v had evolved from a common ancestor which appeared in one focal place in November 1963 +/- 21 months, about 7 years before the first isolation of CA24v in Singapore. The tree also revealed that all the recent epidemic isolates in 1985-1989 including Asian and Ghanian strains diverged from each other after 1981. This finding is consistent with the evidence that AHC due to CA24v had been confined to Southeast Asia and the Indian subcontinent until 1985, then suddenly and explosively spread to other areas where no CA24v isolations had been reported.     

Use of high-frequency ultrasound imaging to improve delineation of anterior uveal melanoma for proton irradiation

The aim of this study was to evaluate high-frequency ultrasound imaging (HFUI) as an aid in localizing anterior margins of tumours of the eye for proton therapy. Proton irradiation of ocular melanoma requires an accurate assessment of all tumour margins. The tumour is marked surgically by suturing to the sclera four or five tantalum rings on the borders of the tumour defined by transillumination. In order to evaluate the clinical usefulness of high-frequency ultrasound imaging, four and five rings were surgically placed in a patient with an iris/ciliary body melanoma and in a patient with ciliochoroidal melanoma using transillumination to localize the tumour margins. Subsequently margins were verified by HFUI. In the first patient, the distances between the rings and the limbus were measured using calipers during surgery and were compared with HFUI measurements and measurements from planning software. The distances were comparable within 0.5 mm. In the second patient the treatment was planned in two different ways using EYEPLAN software. In the first scenario the shape of the tumour and its relation to the rings were obtained from the surgeon's mapping, the fundus drawing using a transilluminating point light source, and the HFUI. In the second scenario, the shape of the tumour was deduced from the ring positions only. It was observed that the maximum difference between the tumour edge as seen on high-frequency ultrasound images and the rings was 2.6 mm. The tumour volume was underestimated by 39% when tumour shape was obtained from ring positions only. During the past year we have utilized HFUI in 18 patients having tumours involving the anterior segment of the eye, among which four were treated with proton therapy. In conclusion, we believe that high-frequency ultrasound imaging provides additional information with respect to the location of tumour margins in ciliary body and anterior uveal melanoma. Occult extension of the tumour within the ciliary body or posterior iris may not be appreciated by transillumination alone.     

Syndiotactic polymerization of styrene with cyclopentadienyltribenzyloxytitanium/methylaluminoxane catalyst

Syndiotactic polystyrene was efficiently prepared in the presence of a homogeneous catalyst composed of (η⁵-cyclopentadienyl)tribenzyloxytitanium and methylaluminoxane (MAO) as a cocatalyst. Influences of various polymerization conditions, e.g., [Ti]. [MAO], [St], temperature and the content of retained trimethylaluminium (TMA) in MAO on the catalytic activity, syndiotacticity and molecular weight of the polymer were studied. It was found that the retained TMA plays an important role in the polymerization. Lowering the retained TMA content in MAO decreases the activity of the catalyst remarkably. Addition of external alkylaluminium (TMA or triisobutylaluminium) into the catalyst system with MAO containing low amounts of retained TMA promotes styrene polymerization.     

Homologous and heterologous neutralization antibody responses after immunization with Japanese encephalitis vaccine among Taiwan children

Because 21 immunized children (13%) among the 162 confirmed Japanese encephalitis (JE) cases during 1986–1991 occurred in Taiwan, we collected 320 serum samples from Taiwan children aged 15–31 and 27–44 months immediately before the 1st dose (n = 41) and 1–3 months after the 2nd dose (n = 78, 27 pairs), and immediately before (n = 58) and 1–3 months after the 3rd dose (n = 143,44 pairs) to determine neutralization antibody (Nt Ab) against the Nakayama (N) and Beijing-1 (B) strains and two Taiwan wild type JE viruses (JEV): CC-27 and CH-1392. Our Nt results showed that (1) B vaccine stimulated a better homologous Ab response than N vaccine for Nt Ab seropositivity rate (NASR), produced a higher level of Nt titer after the primary immunization [2 doses = 100% vs. 91%, geometric mean titer (GMT) = 115 vs. 22], had a greater booster effect (3 doses: 100% vs. 95%; GMT = 320 vs. 33), and showed a better capability to neutralize two local Taiwan JEV strains, particularly only after 3 doses (ave. NASR for B vs. N = 90% vs. 10%; and GMT for B vs. N = 154 vs. 1); (2) the two wild type JEV strains had different plaque morphology and antigenic variation and the CC-27 strain was not neutralized as well as the CH-1392 strain after 3 doses of vaccine (BBB or NNN or NNB); and (3) 30% of the children had lost JEV Nt Ab one year after the 2nd dose of N vaccine and natural infection with JE virus did occur among those children after immunization. In conclusion, (1) three doses of mouse-brain vaccine are the minimum requirement to protect children against the local Taiwan JEV; (2) the best strain for a JE vaccine depends on level of Nt Ab it induced, the molecular epidemiology and antigenic variation of the JEV in each local area; and (3) future vaccine must produce better B- and T-cell memory. © 1994 Wiley-Liss, Inc.     

c-kit Expression in small cell carcinoma of the urinary bladder: prognostic and therapeutic implications.

The prognosis for small cell carcinoma of the urinary bladder is poor, and strategies for improved therapy are needed. Targeted therapy against the c-kit proto-oncogene has been successful in the management of gastrointestinal stromal tumor. We investigated the expression of c-kit in 52 cases of small cell carcinoma of the urinary bladder. Specimens with more than 10% of cells demonstrating strong membrane staining were considered to have positive immunostaining for c-kit. c-kit expression was detected in 21 of 52 specimens from these patients. Among the 21 specimens, seven had less than 10% staining, and were considered to be negative. Nine had 11-50% staining, and five had more than 50% staining. Overall, 14 of 52 (27%) small cell carcinomas of the urinary bladder were positive for c-kit expression. During a median follow-up of 11 months, 60% of the patients died of bladder cancer. No association was found between c-kit expression and survival or other clinicopathologic parameters. Five-year cancer-specific survivals for c-kit-positive and c-kit-negative tumors were 9 and 15%, respectively (P=0.36). A significant proportion (27%) of small cell carcinomas of the urinary bladder expressed c-kit, suggesting that it may prove useful as a therapeutic target in small cell carcinoma of the urinary bladder.     

Electrophysiological neurodifferentiation of subventricular zone-derived precursor cells following stroke

Stroke in rodents is associated with increased neurogenesis and the migration of newborn neurons to sites of brain ischemia, where they may participate in repair and recovery. To determine if neurogenesis following stroke yields functional new neurons, we labeled neuronal precursors in the mouse subventricular zone (SVZ) with a lentivirus-green fluorescent protein vector, produced stroke by occluding the middle cerebral artery, and detected newborn neurons 8 weeks later by fluorescence microscopy. Patch-clamp studies on fluorescent neurons in the cortical region surrounding infarction showed tetrodotoxin-sensitive Na⁺ action potentials and spontaneous excitatory post-synaptic currents, suggesting that ischemia led to functional neurogenesis with synaptic integration. These findings support the hypothesis that enhancing endogenous neurogenesis after stroke might have therapeutic benefit.     

Taste receptor cells express pH-sensitive leak K+ channels

Two-pore domain K+ channels encoded by genes KCNK1-17 (K2p1-17) play important roles in regulating cell excitability. We report here that rat taste receptor cells (TRCs) highly express TASK-2 (KCNK5; K2p5.1), and to a much lesser extent TALK-1 (KCNK16; K2p16.1) and TASK-1 (KCNK3; K2p3.1), and suggest potentially important roles for these channels in setting resting membrane potentials and in sour taste transduction. Whole cell recordings of isolated TRCs show that a leak K+ (Kleak) current in a subset of TRCs exhibited high sensitivity to acidic extracellular pH similar to reported properties of TASK-2 and TALK-1 channels. A drop in bath pH from 7.4 to 6 suppressed 90% of the current, resulting in membrane depolarization. K+ channel blockers, BaCl2, but not tetraethylammonium (TEA), inhibited the current. Interestingly, resting potentials of these TRCs averaged -70 mV, which closely correlated with the amplitude of the pH-sensitive Kleak, suggesting a dominant role of this conductance in setting resting potentials. RT-PCR assays followed by sequencing of PCR products showed that TASK-1, TASK-2, and a functionally similar channel, TALK-1, were expressed in all three types of lingual taste buds. To verify expression of TASK channels, we labeled taste tissue with antibodies against TASK-1, TASK-2, and TASK-3. Strong labeling was seen in some TRCs with antibody against TASK-2 but not TASK-1 and TASK-3. Consistent with the immunocytochemical staining, quantitative real-time PCR assays showed that the message for TASK-2 was expressed at significantly higher levels (10-100 times greater) than was TASK-1, TALK-1, or TASK-3. Thus several K2P channels, and in particular TASK-2, are expressed in rat TRCs, where they may contribute to the establishment of resting potentials and sour reception.     

Urea permeation and hydrolysis through hollow fiber dialyzer immobilized with urease

The surface of commercial polyacrylonitrile hollow fibers was hydrolyzed and covalently bonded with urease by using glutaraldehyde. Mini-modules assembled with these modified hollow fibers were then used to study the effect of concentration of glutaraldehyde, pH, and temperature on the catalysis of urea. The immobilized amount increased with the concentration of glutaraldehyde. However, urease immobilized with 5% glutaraldehyde had higher activity (0.38 micromol/min/mg-urease) than with other concentrations. The activity of the immobilized urease remained at above 0.32 micromol/min/mg-urease over wider applicable ranges of temperature (4-70 degreesC) and pH (5-8) compared to free urease. For comparison, the activity of immobilized urease was 0.35 micromol/min/mg-urease, while that for non-immobilized urease was 0.33 micromol/min/mg-urease at pH 7 and 20 degreesC. The removal of urea using urease-immobilized dialyzer was demonstrated with in-vitro dialysis and showed faster removing rate of urea than a regular dialyzer by 2.7 times.     

慢性乙肝患者杀伤性免疫细胞功能的研究

通过对４４例病毒性肝炎患者Ｔ细胞亚群，ＮＫ细胞活性与ＬＡＫ细胞活性的观察，探讨了在慢性乙型肝炎病毒复制与非复制状态下的杀伤性细胞活性。结果表明：在乙肝病毒的高复制状态下，ＣＤ８＾＋细胞数增加，ＣＤ４＾＋／ＣＤ８＾＋比例显著下降；ＮＫ细胞活性与ＬＡＫ细胞活性也明显低下，且在ＨＢｅＡｇ与ＨＢＶＤＮＡ阳性组中，ＮＫ活性与ＬＡＫ活性的改变与ＨＢｅＡｇ的Ｐ／Ｎ值变化呈显著负相关，而ＮＫ活性与ＬＡＫ活性变化则