Low rates of antigen detection and virus isolation from the peripheral blood leukocytes of dengue fever patients

We evaluated direct fluorescent antibody (FA) testing of peripheral blood leukocytes (PBL) from patients in Puerto Rico with serologically and/or virologically confirmed dengue fever as a possible rapid diagnostic test and compared rates of dengue virus isolation from PBL with the rates from plasma or serum using the mosquito inoculation technique. Dengue antigen was detected in the PBL of only 1 of 19 patients with confirmed dengue. Virus was isolated from 3 of 19 PBL specimens and from 6 of 19 acute-phase serum or plasma samples. Four viruses were obtained from serum or plasma only and 1 isolate came from PBL only. We conclude that FA testing of PBL from dengue fever patients has little promise as a rapid diagnostic technique. Despite small numbers, our data suggest that virus isolation from PBL is less sensitive than that from serum or plasma. Our results differ considerably from those of previous studies of dengue hemorrhagic fever patients conducted in Thailand.     

Excellent outcome of stage II neuroblastoma is independent of residual disease and radiation therapy

The optimal management for patients with stage II neuroblastoma has not yet been established. In order to determine the impact of adding chemotherapy and/or radiation therapy to surgery, we reviewed by questionnaire 156 patients with stage II neuroblastoma treated by 28 Childrens Cancer Study Group (CCSG) institutions from 1978 to 1985. Survival and progression-free survival (PFS) were analyzed by life-table methods with respect to age at diagnosis, site and size of primary tumor, spinal cord involvement, extent of initial resection, and treatment in addition to surgery. The overall 5-year survival was 96%; the PFS was 90%, similar to previous CCSG studies. Age at diagnosis had a small impact on PFS, with 92% PFS for patients less than 2 years of age at diagnosis, and 84% for those greater than 2 (P = .10). The only site with an adverse outcome was the head and neck (n = 11), with a PFS of 68% compared with 93% for the remaining sites (P = .02). Size of primary and intraspinal extension of primary did not affect PFS. The extent of resection and subsequent treatment with radiation therapy and/or chemotherapy did not affect the PFS. The outcome for 75 patients treated with surgery alone (6-year PFS, 89%) was not significantly different from that of 66 patients receiving radiation therapy (6-year PFS, 94%). There was no significant difference between 40 patients with gross or microscopic residual disease treated with surgery alone (PFS, 92%) and 59 patients with residual disease who also received radiation (PFS, 90%). Five of seven patients who progressed after surgery alone have been salvaged with further therapy and are now free of disease. One survives with disease, so that the 6-year survival is 98% for those treated initially with surgery alone, compared with 95% for those receiving radiation therapy and/or chemotherapy. These data suggest that surgery alone, even if complete resection is not achieved, is sufficient initial therapy for stage II neuroblastoma. The data also identify another stage of neuroblastoma, in addition to stage IV-S, for which almost all patients have a favorable prognosis because their tumor may be biologically limited in growth.     

槐定碱的气相色谱测定法及其在兔体内的药代动力学

兔血浆中槐定碱的气相色谱测定条件:氮气、氢气和空气的流速分别为60,40和380ml/min。填充柱、进样室和检测室温度分别为190,280和300℃。内标物为槐胺碱,血浆中槐定碱浓度为3.0和10.0μg/ml时,测得回收率分别为103.6±2.4%(CV=2.34%)和101.5±6.5%(CV=6.35%)。线性范围为1～15/μg/ml。槐定碱在兔体内的药代动力学按二房室拟合,其混合参数值:α.0.0864 min^-1;β.0.00932 min^-1;Vc.1.19 L/kg;V_area.2.07 L/kg;V_dvv.1.99L/kg;Cl.19.3 ml/min·kg;MRT.103min。     

部分甲基化葡萄糖的制备

以葡萄糖、甲基葡萄糖苷或可溶性淀粉为原料,适当控制反应条件进行Haworth⁽¹⁾甲基化可得到一系列不同取代数目及不同取代位置的甲基化葡萄糖(苷)。经反相HPLC分离制备后得到纯品。各产物都用气相色谱、质谱作了结构鉴定。对甲基化反应条件和HPLC分离条件进行了讨论。此法适合于实验室小量制备单糖或其甲基糖苷的甲基化标准物,供糖缀合物结构鉴定之需。     

Oligonucleotide probes to the 16s ribosomal RNA: implications of sequence homology and secondary structure with particular reference to the oral species Prevotetla intermedia and Prevotetla nigrescens

Eight oligonucleotides based upon regions of the small subunit 165 ribosomal RNA gene sequences were analysed against a background of their position within the molecule and their two-dimensional structure to rationalise their use in recognising Prevotetla intermedia and Prevotetla nigrescens . The 41 clinical isolates from both oral and respiratory sites and two reference strains were subjected to DNA-DNA hybridisation and multilocus enzyme etectrophoresis to confirm their identity. Alignment of oligonucleotide probes designated I Bi-2 to I Bi-6 (for P. intermedia ) and 2Bi-2 (for P. nigrescens ) with the 165 rRNA suggested that these probes lacked specificity or were constructed from hypervariable regions. A 52– mer oligonucleotide (designated Bi) retiably detected both species. Because of the high degree of concordance between the I 65 rRNAs of both species, it was necessary to vary the stringency of hybridisation conditions for detection of both species. Thus probe I Bi-I recognised P. intermedia while I Bi-l detected both P. intermedia and P. nigrescens at low stringency. However, under conditions of high stringency only P. nigrescens was recognised by probe 2Bi-I. These probes were highly specific and did not hybridise with DNA from the closety retated P. corporis , nor other periodontal pathogens such as Fuso-bacterium nucleatum, Actinobacillus actinomycetemcomit-ans , Treponema denticola and several pigmented species such as Prevotetla metoninogenica, P. denticola, P. loescheii, Porphyramonas osaccharolytica, Py. endodontalis, Py. gin-givalis, Py . levii, and Py. macacae .     

Evaluation of the Shimada classification in advanced neuroblastoma with a special reference to the mitosis-karyorrhexis index: a report from the Childrens Cancer Study Group

Histopathology of 46 cases of Stage III and Stage IV neuroblastoma collected at the Childrens Cancer Study Group Pathology Center was reviewed independently by two pathologists to formulate essential steps in the determination of the mitosis-karyorrhexis index (MKI), to assess interobserver concordance, and to evaluate prognostic significance of the Shimada classification system. "Absolute" agreement (obtained after the review by both pathologists) or "consensus" agreement (obtained after the second review of cases with initial discrepancy) was achieved with respect to category of MKI in 93% of 40 stroma-poor neuroblastomas and prognostic subgrouping in 98% of the total cases. This classification system distinguished favorable from unfavorable prognostic subgroups significantly (P = 0.0002), and the prognostic effects were largely unaltered when adjusted for age and stage of the patients (P = 0.0005) in this series.     

Revised and updated nomenclature for highly pathogenic avian influenza A (H5N1) viruses

The divergence of the hemagglutinin gene of A/goose/Guangdong/1/1996‐lineage H5N1 viruses during 2011 and 2012 (807 new sequences collected through December 31, 2012) was analyzed by phylogenetic and p‐distance methods to define new clades using the pre‐established nomenclature system. Eight new clade designations were recommended based on division of clade 1·1 (Mekong River Delta), 2·1·3·2 (Indonesia), 2·2·2 (India/Bangladesh), 2·2·1·1 (Egypt/Israel), and 2·3·2·1 (Asia). A simplification to the previously defined criteria, which adds a letter rather than number to the right‐most digit of fifth‐order clades, was proposed to facilitate this and future updates.     

The identification of urinary bile alcohols by gas chromatography–mass spectrometry in patients with liver disease and in healthy individuals

Abstract. The neutral steroid fractions in the urine of eleven patients suffering from various forms of liver disease with cholestasis and of ten healthy individuals were studied by glass capillary gas chromatography-mass spectrometry. The steroid conjugates in urine were enzymatically solvolysed, the liberated steroids extracted and transformed into the trimethylsilylether for measurements.
The excretion rates of androstane and pregnane metabolites of patients with liver disease were far lower than those of healthy persons. The main compounds in the urine of the former were the bile alcohols 27 - nor -3α, 7α, 12α, 24, 25 - pentahydroxy - 5β - cholestane and 3α, 7α, 12α, 25, 26 - pentahydroxy - 5β - cholestane. Our data suggest a correlation between the excretion rates of these bile alcohols and the serum levels of bilirubin. While the excretion rate of the two bile alcohols in the urine of healthy individuals was approximately 0.24 mg/24 h (0.6 μmol/24 h) a patient with a serum bilirubin of 841 μmol/1 excreted 4 mg/24 h (9 μmol/24 h). The accumulation of bile alcohols described in this study possibly indicates alternative pathways of cholic acid formation in liver disease.