Flow Cytometric Analysis of Cellular Changes in Mice after Intradermal Inoculation with a Liposome–Iscom Adjuvanted Vaccine

As it is not known what changes to leucocyte homeostasis are mandatory for effective adjuvant action, the biological relevance of systemic changes elicited by different vaccine formulations can only be interpreted in the context of the immunological outcomes. We used flow cytometry to quantify the changes in leucocyte subsets induced in mice intradermally immunized with SAMA4 (adjuvant group), outer membrane proteins (OMP) purified from Actinobacillus pleuropneumoniae (OMP antigen group), SAMA4 adjuvanted OMP (OMP vaccine group), or phosphate-buffered saline (PBS: control group). This approach allowed direct comparisons to be made between the effects of antigen, adjuvant or antigen–adjuvant complexes on immune effector cell populations. Antigens complexed with the liposome–iscom hybrid adjuvant, SAMA4, generated strong antibody responses and cytotoxic T-cell activity in animals immunized intradermally, reflecting remobilization and recruitment of specific cell populations. Splenomegaly, due to granulocytosis, monocytosis and megakaryocytosis, was most prominent in the OMP vaccine group. Histological examination of spleen sections confirmed that these changes were due primarily to splenic haematopoiesis. Circulating numbers of granulocytes and monocytes increased significantly (P < 0.05) in the blood of the OMP vaccine group, as did granulocyte numbers in the lungs (P < 0.05). No changes in T- and B-cell numbers were detected by flow cytometry in the spleens, lungs or blood over the 28-day period in any treatment group. Thymocyte numbers (predominantly CD4⁺CD8⁺ cells) in the OMP vaccine group fell by 95% within 3 days of immunization. Identical cellular responses were obtained when an innocuous antigen, ovalbumin, was complexed with SAMA4 instead of OMP, thus demonstrating that the adjuvant effects of SAMA4 were due to synergistic interaction between antigen and adjuvant and not due to the presence of toxic components. The association of strong adaptive immune responses with such complex changes in leucocyte homeostasis induced by complexing adjuvant and antigen suggested that the changes were important for effective vaccination and were not purely circumstantial.     

Chlorofluorocarbon CFCs, Potential Alternative HCFCs and HFCs, and Related Chlorinated Compounds: Mass Spectral Study, Part II

In a previous publication, mass spectrometry was applied to the study of several ions formed by dissociative ionization of ethanes, partially substituted with fluorine and chlorine, and to the determination of the fragmentation pattern of each related compound. The aim of the present investigation was to extend this mass spectral study (70 eV and low ionization energy) to a group of closely related analogs, hydrofluorocarbons (HCFCs), HFCs, chlorinated fluorocarbons, and chlorinated hydrocarbons—halogenated methane, ethane, and propane molecules—to make an attempt to predict the relative bond strength and then the behavior—metabolic pathway and tropospheric degradation—of new HCFCs and HFCs.     

以酶解法从中药及药渣制备β—葡萄糖的研究

本实验首次将工业纤维素酶应用于中药及药渣中，探讨利用工业纤维素酶使中药及药渣的纤维素酶解为β－葡萄糖；实验结果表明：在５０℃，ｐＨ４．５，底物浓度为１５％的条件下，４８小时可酶解得５％以上的β－葡萄糖。     

Increases in plasma beta-endorphin concentrations during exercise do not contribute to increases in heart rate following autonomic blockade in man.

1. Intrinsic heart rate (IHR: heart rate following autonomic blockade with atropine and propranolol) increases with exercise. The opioid antagonist naloxone has been shown to decrease IHR at rest, raising the possibility that increases in IHR with exercise are beta-endorphin related, since beta-endorphin concentrations have also been shown to rise during exercise. 2. We examined the effects of naloxone (10 mg) on IHR and plasma beta-endorphin levels during aerobic exercise in eight healthy, male subjects in a single blind, crossover study. 3. IHR increased with 25 min bicycling from 97.1 +/- 1.4 to 129.7 +/- 1.2 beats min-1 (mean +/- s.e. mean). This rise was not affected by administration of naloxone. 4. Plasma beta-endorphin concentration rose from 31.1 +/- 3.8 to 94.9 +/- 23.9 pg ml-1 after 25 min exercise. This exercise-induced rise in beta-endorphin concentration was further increased (P less than 0.05) in the presence of naloxone. 5. Our results confirm a rise in IHR and beta-endorphin concentrations with acute exercise but indicate that the changes in IHR are not endorphin-related.     

早期自体角膜缘移植术治疗碱烧伤

目的探讨角膜缘移植术在眼碱烧伤中的治疗意义。方法新西兰兔１４只，１．０ｍｏｌ／ＬＮａＯＨ溶液眼前节烧伤１分钟。其中７只兔于烧伤后即实施自体角膜缘移植术，另７只兔为对照。术后观察１个月，取其角膜进行光镜及扫描电镜观察。结果手术组角膜上皮愈合快，角膜新生血管及角膜炎细胞浸润均较对照组轻。结论早期自体角膜缘移植术治疗碱烧伤可加速角膜上皮愈合、抑制角膜新生血管形成、减轻角膜炎症反应及预防角膜溃疡的发生。     

Oral immunization of mice with attenuated Salmonella typhimurium aroA expressing a recombinant Mycoplasma hyopneumoniae antigen (NrdF).

Mycoplasma hyopneumoniae is the etiological agent of porcine enzootic pneumonia, a commercially expensive respiratory disease of swine. Salmonella typhimurium SL3261 was used as a live carrier of plasmid pKF1, which encodes a 15-kDa recombinant M. hyopneumoniae protein. This expressed recombinant protein consists of the carboxy-terminal 11 kDa of a 42-kDa M. hyopneumoniae NrdF ribonucleotide reductase R2 subunit protein. Rabbit anti-15-kDa serum was able to inhibit the growth of viable M. hyopneumoniae J in vitro. When used as a live oral vaccine, S. typhimurium SL3261(pKF1) induced a significant secretory immunoglobulin A immune response in the lungs of mice orally immunized against the M. hyopneumoniae antigen. Utilization of live oral vaccines expressing potentially protective M. hyopneumoniae proteins, such as the NrdF antigen, which can stimulate a lung mucosal response against surface-accessible proteins may provide a cost-effective alternative to the present control strategies used for porcine enzootic pneumonia.     

The effect of pre-vertebroplasty tumor ablation using laser-induced thermotherapy on biomechanical stability and cement fill in the metastatic spine

A biomechanical study comparing simulated lytic vertebral metastases treated with laser-induced thermotherapy (LITT) and vertebroplasty versus vertebroplasty alone. To investigate the effect of tumor ablation using LITT prior to vertebroplasty on biomechanical stability and cement fill patterns in a standardized model of spinal metastatic disease. Vertebroplasty in the metastatic spine is aimed at reducing pain, but is associated with risk of cement extravasation in up to 10%. Six pairs of fresh-frozen cadaveric thoracolumbar spinal motion segments were tested in axial compression intact, with simulated metastases and following percutaneous vertebroplasty with or without LITT. Canal narrowing under load, pattern of cement fill, load to failure, and LITT temperature and pressure generation were collected. In all LITT specimens, cement filled the defect without extravasation. The canal extravasation rate was 33% in specimens treated without LITT. LITT and vertebroplasty yielded a trend toward improved posterior wall stability (P = 0.095) as compared to vertebroplasty alone. Moderate rises in temperature and minimal pressure generation was seen during LITT. In this model, elimination of tumor by LITT, facilitates cement fill, enhances biomechanical stability and reduces the risk of cement extravasation.     

Usefulness of contrast material in MR of patients with neurofibromatosis type 1.

PURPOSEOur objective was to determine the usefulness of routine administration of contrast material in brain MR imaging for the evaluation of areas of probable myelin vacuolization and neoplasms in patients with neurofibromatosis type 1 (NF-1).METHODSWe retrospectively reviewed 112 consecutive contrast-enhanced brain MR studies obtained over a period of 7 years in 109 symptomatic and asymptomatic patients compiled from two institutional NF-1 data bases. MR studies were analyzed for areas of probable myelin vacuolization, with attention to degree of enhancement and its impact on lesion detection and characterization. Usefulness of contrast material was graded as 0 = not useful, 1+ = somewhat useful, and 2+ = useful.RESULTSOf 112 studies, 45% (n = 49) were normal. In the remaining 63 studies, 88 regions of probable myelin vacuolization and 52 tumors were identified. Enhancement was not observed in any regions of probable myelin vacuolization. Enhancement was present in 31% of tumors, and, of these, was found to be useful in 44%, somewhat useful in 12%, and not useful in 44%. For enhancing tumors, contrast agent was useful for lesion detection in 19% and for lesion characterization in 25%.CONCLUSIONContrast administration is useful in baseline MR studies to maximize tumor detection and characterization, to add confidence to the diagnosis of benign probable myelin vacuolization, and to document stability of neoplasms on follow-up examinations.     

Anti-tumor effect of tumor necrosis factor and its induction of tumor variant of MBT-2 transitional cell carcinoma of the bladder

The effect of tumor necrosis factor (TNF), a potential anti-tumor agent, was assessed both in vivo and in vitro against MBT-2 transitional cell carcinoma of the bladder in C3H/HeHa mice. Systemic administration of either single or multiple doses of TNF into tumor bearing animals resulted in partial tumor regression and had a consistent but transient anti-tumor effect. Compared to control, untreated tumor bearing animals, TNF-treated tumor bearing mice had significantly smaller tumor volumes and slower tumor growth rates over the period of 12 days following TNF inoculation. No significant difference in tumor volumes and tumor growth rates between controls and TNF-inoculated mice was observed from day 12 to day 21 after TNF treatment. Assessment of TNF cytotoxicity against in vitro MBT-2 cell line using 3[H]-thymidine proliferation assay showed significant sensitivity of MBT-2 cells to treatment with TNF. A "Variant" MBT-2 cell line was derived by sequential culturing of the original MBT-2 cells in the presence of progressively higher concentrations of TNF in culture medium. Although the significant growth suppression on the MBT-2 tumor appears to be transient, further studies are warranted which may elucidate the immunologic and biologic behavior of TNF and this transplantable animal tumor.     

β-Methyl-15-p-iodophenylpentadecanoic acid metabolism and kinetics in the isolated rat heart

The use of 15-p-iodophenyl--methyl-pentadecanoic acid (Me-IPPA) as an indicator of long chain fatty acid (LCFA) utilization in nuclear medicine studies was evaluated in the isolated, perfused, working rat heart. Time courses of radioctivity (residue curves) were obtained following bolus injections of both Me-IPPA and its straight chain counterpart 15-p-iodophenyl-pentadecanoic acid (IPPA). IPPA kinetics clearly indicated flow independent impairment of fatty acid oxidation caused by the carnitine palmitoyltransferase I inhibitor 2[5(4-chlorophenyl)pentyl]oxirane-2-carboxylate (POCA). In contrast, Me-IPPA kinetics were insenstive to changes in fatty acid oxidation rate and net utilization of long chain fatty acid. Analysis of radiolabeled species in coronary effluent and heart homogenates showed the methylated fatty acid to be readily incorporated into complex lipids but a poor substrate for oxidation. POCA did not significatly alter metabolism of the tracer, suggesting that the tracer is poorly metabolized beyond Me-IPPA-CoA in the oxidative pathway.