Analysis of recombinant human tumour necrosis factor-alpha-induced CD4 expression on human eosinophils.

We examined the hypothesis that one of the pro-inflammatory cytokines, tumour necrosis factor-alpha (TNF-alpha), could induce expression of the adhesion molecule CD4 on human eosinophils. We further examined the effector function of CD4 and the mechanisms regulating CD4 expression. Human eosinophils were cultured with various concentrations of recombinant human TNF-alpha (rhTNF-alpha) with or without various drugs for 24 hr. After culture, eosinophils were stained for CD4 using a monoclonal antibody and then analysed by flow cytometry. Eosinophil-derived neurotoxin (EDN) release as eosinophil degranulation was examined by cross-linking of CD4 on eosinophils. The rhTNF-alpha induced CD4 expression on human eosinophils in a dose- and time-dependent fashion; rhTNF-alpha-induced CD4 expression was significantly inhibited by 10(-6) M cycloheximide, 10(-8) M dexamethasone, or 10(-6) M herbimycin A. Recombinant human interferon-gamma inhibited rhTNF-alpha-induced CD4 expression in a dose-dependent manner. However, cross-linking of CD4 on eosinophils did not evoke EDN release, suggesting that newly expressed CD4 molecules on human eosinophils do not play any role in triggering degranulation. Our data indicate that TNF-alpha-induced CD4 expression on human eosinophils is dependent on protein synthesis and may be dependent on tyrosine kinase activity.     

Flavonoids such as luteolin, fisetin and apigenin are inhibitors of interleukin-4 and interleukin-13 production by activated human basophils

BACKGROUND: We have previously shown that fisetin, a flavonol, inhibits IL-4 and IL-13 synthesis by allergen- or anti-IgE-antibody-stimulated basophils. This time, we investigated the inhibition of IL-4 and IL-13 production by basophils by other flavonoids and attempted to determine the fundamental structure of flavonoids related to inhibition. We additionally investigated whether flavonoids suppress leukotriene C4 synthesis by basophils and IL-4 synthesis by T cells in response to anti-CD3 antibody. METHODS: Highly purified peripheral basophils were stimulated for 12 h with anti-IgE antibody alone or anti-IgE antibody plus IL-3 in the presence of various concentrations of 18 different kinds of flavones and flavonols. IL-4 and IL-13 concentrations in the supernatants were then measured. Leukotriene C4 synthesis was also measured after basophils were stimulated for 1 h in the presence of flavonoids. Regarding the inhibitory activity of flavonoids on IL-4 synthesis by T cells, peripheral blood mononuclear cells were cultured with flavonoids in anti-CD3-antibody-bound plates for 2 days. RESULTS: Luteolin, fisetin and apigenin were found to be the strongest inhibitors of both IL-4 and IL-13 production by basophils but did not affect leukotriene C4 synthesis. At higher concentrations, these flavonoids suppressed IL-4 production by T cells. Based on a hierarchy of inhibitory activity, the basic structure for IL-4 inhibition by basophils was determined. CONCLUSIONS: Due to the inhibitory activity of flavonoids on IL-4 and IL-13 synthesis, it can be expected that the intake of flavonoids, depending on the quantity and quality, may ameliorate allergic symptoms or prevent the onset of allergic diseases.     

Organ transplantation and laboratory tests

The media has recently been featuring organ transplantation from various viewpoints. Furthermore, Novel Prizes 1990 for Medical & Physiological fields were awarded to Drs. JE Murray and ED Thomas, both pioneers of clinical transplantation. Our topic has been timely indeed. This symposium mainly dealt with laboratory tests vs. various types of organ transplantation. In reality though, only kidney and bone marrow transplantations have been practiced in Japan; thus, Dr. I Yokoyama, University of Pittsburgh, discussed liver transplantation. First, Dr. K Uchida lectured on the recent advancement of immunosuppressive drugs and improvement in the clinical outcome of kidney transplantation. Serum creatinine determination is the only parameter for rejection besides renal biopsy. Drs. K Miyamura & Y Morishima discussed about PCR method to detect MRD (minimal residual diseases). There are positive relationships between the remaining leukemic cells and the relapse of leukemia even though the patients are in clinical remission. Dr. H Funada dealt with the importance of "sterile room treatment" for bone marrow transplantation. It protects patients from infection, minimizes GVHD and prolongs survival time after transplantation. Dr. Yokoyama stressed the importance of back-up system, i.e. drug-monitoring, coagulation tests, pathological examination, biochemical tests, blood transfusion services for successful liver transplantations. Dr. T Fukunishi discussed the importance of developing the organ donor and coordinator system to promote kidney transplantation from cadaver. He also dealt with virus antibody tests for selecting donors. All discusssions stressed on the importance of the 24-hour laboratory back-up system performing emergency tests but no specific laboratory test for organ transplantation was necessary.     

Usefulness of "essential laboratory tests" for the establishment of the initial diagnosis in new outpatients]

"Essential laboratory tests" advocated by Japan Society of Clinical Pathology were simultaneously performed with the history taking of the present illness and the physical examination in 1,026 new patients visited the outpatient unit of Comprehensive Medicine, National Defense Medical College. We have analyzed the usefulness of the "essential laboratory tests" for the establishment of the initial diagnosis in evaluable 750 patients by comparing the diagnosis made only by the history taking and the physical examination (tentative initial diagnosis) with that included the results of these laboratory tests. The "essential laboratory tests" had contributed to remarkably increased incidences of metabolic and endocrine diseases, liver or biliary tract diseases, renal and urinary tract diseases and anemia after the application of these tests. The initial diagnoses of 61 patients were successfully established by the addition of the "essential laboratory tests" among 157 cases of which tentative initial diagnoses remained undetermined. These tests confirmed the tentative diagnoses in 78 patients, while 78 tentative diagnoses were negated and corrected after evaluation of the results of these tests. Furthermore, the diagnostic tests such as CRP and leukocyte count were useful for the estimation of the nature or degree of seriousness of the disease in 57 patients. In addition, other diseases not related to the patient's chief complaint could be detected in 238 patients (303 total number of diseases) by these tests. These results indicate the usefulness of the "essential laboratory tests" not only for the establishment of more accurate initial diagnosis but also for the screening of the "hidden" diseases such as hyperlipidemia and liver dysfunction.     

Effects of mechanical trauma of blood cells on dynamic viscoelasticity of blood during clotting

Effects of mechanical trauma of platelets and red blood cells on dynamic viscoelasticity of blood during clotting were examined. Two different methods were attempted to give the mechanical damage to blood cells. In one method, a flow apparatus consisting of a thin teflon tube and two sample reservoirs connecting to both ends of the tube was employed. After a blood sample repeated the shuttle flow many times through the tube, the dynamic rigidity modulus (G') and loss modulus (G") of the blood sample during clotting were measured. The mechanical trauma of platelets brought about the increase of G' and G" for clots of PRP and whole blood, in addition to shorter clotting time. Slight hemolysis of red blood cells (less than 0.5% of total hemolysis) only yielded shorter clotting time for whole blood. In other method, packed red blood cells were hemolyzed by exposing the ultrasonic wave (the amount of hemolysis was about 5% of total hemolysis). Addition of supernatant of hemolyzed red blood cells to PRP caused the platelet aggregation. Furthermore, a marked hemolysis brought about higher values of G' and G" for blood clot, in addition to shorter clotting time. These results suggest that the release of constituents from damaged red blood cells and alteration of cell surface due to mechanical damage would accelerate the coagulation sequence.     

Validation and reliability of a Japanese version of the Shoulder Pain and Disability Index: A cross-sectional study

BackgroundThe Shoulder Pain and Disability Index (SPADI) is a simple disease specific questionnaire that is used to evaluate the impact of shoulder disorders. The purpose of this study was to translate the SPADI into Japanese (SPADI-Jp) and evaluate its reliability and validity in Japanese patients with shoulder disorders.MethodsCross-cultural adaptation of the SPADI was performed according to international guidelines. A total of 100 patients with shoulder disorders participated in this study. Each participant was asked to finish the SPADI-Jp, Disability of Arm, Shoulder and Hand (DASH), and the Short-Form 36 (SF-36) at the initial visit. Thirty-four patients repeated the SPADI-Jp to assess the test–retest reliability. The test–retest reliability was quantified using the interclass correlation coefficient (ICC), while Cronbach's alpha was calculated to assess the internal consistency. The construct validity was assessed using Spearman's rank correlation coefficients.ResultsInternal consistency in the SPADI-Jp was very high (0.969), as measured by the Cronbach's alpha. The ICC of the SPADI-Jp was 0.930. There was a strong, positive correlation between the DASH and the SPADI-Jp (r = 0.837, p < 0.001). The SPADI-Jp was significantly correlated with most of the SF-36 subscales. The correlations of the SPADI-Jp with physical subscales of the SF-36 were stronger than those with the other subscales.ConclusionsWe demonstrated that the SPADI-Jp is a reliable and valid self-assessment tool. Because cross-cultural adaptation, validation, and reliability of the disease-specific questionnaire for shoulder pain and disability have not been evaluated in Japan, the SPADI-Jp can be useful for evaluating such patients in the Japanese population.     

Acoustic evaluation of postoperative hoarseness in adult patients

The present study was performed to evaluate postoperative hoarseness quantitatively by means of acoustic wave form analysis. Pitch and amplitude perturbation (PPQ, and APQ), and normalized noise energy (NNE) were measured along with the frequency characteristics in 51 adult patients undergoing elective surgery. The normal values for these acoustic parameters were less than 0.5%, less than 2.0%, and less than -10 dB, respectively. Vowel sound "E" was recorded and evaluated before the induction of anesthesia and on the morning of the day after the surgery. PPQ increased from 0.39% to 1.00% (P less than 0.05), APQ increased from 3.34% to 6.62% (P less than 0.05), and NNE increased from -9.19 dB to -4.74 dB (P less than 0.05). Eighteen percent of the patients showed abnormal values in all parameters preoperatively, but 45% of the patients postoperatively (P less than 0.05). These findings suggest that even the short term intubation resulted in the postoperative hoarseness, and this method is a useful and non-invasive bed-side test to evaluate postoperative hoarseness quantitatively.     

Heterozygote effects in dreher mice

The dreher mutation (gene symbol: dr) is an autosomal recessive mutation located on chromosome 1 of the mouse. Homozygous dreher mice (dr/dr) are ataxic, have a white belly spot, a short-tail, inner ear and skeletal malformations, and a variety of CNS abnormalities. Recently in our dreher colony (the drsst-J allele on a B6C3Fe background), we noticed mice with one or more white belly spots typical of drsst-J/drsst-J mice but which were non-ataxic and had a normal tail length; wild-type mice (+/+) of the same genetic background do not have simialr belly spots. Results of three breeding experiments indicate that a new mutation had not occurred, but rather that the spotted, non-ataxic mice are heterozygous dreher mice (drsst-J/+). Histological examination showed that drsst-J/+ mice have abnormalities in the hippocampal formation that are qualitatively similar to those found in drsst-J/drsst-J mice. Most frequently there is an increase in the number of pyramidal cells in CA3 and a marked thickening of the pyramidal cell layer. In contrast to dreher homozygotes the cerebellum appears to have a normal foliation pattern and no discernible laminar abnormalities. Thus, both breeding experiments and histological examination indicate that drsst-J is semidominant. We speculate that drsst-J is a "loss of function" mutation, but, in any event, the presence of phenotypic abnormalities in drsst-J/+ mice may be useful in identifying the primary developmental defect in dreher mice.     

Proffered opinion forum medical decision making, impact on laboratory tests

Raising medical expenditure is a universal problem. The Japanese government started taking measures not to increase medical expenses in the recent years. Laboratory tests are no exception. Therefore we as clinical pathologists have reevaluated laboratory tests from the primary care situations to more advanced stages of clinical medicine. As the first step, we formed a subcommittee to effective by utilize laboratory tests and agreed to establish the "essential lab tests". Essential lab tests are made of qualitative bed side tests (type I) and slightly advanced tests (type II) and these tests are to be done on every patient at their first clinic visit. These tests are also to be performed parallel to physical examination and history-taking. The results revealed 50% and 90% positivities on the type I and type II tests, which indicate rather effective means for making an initial impression.