Evaluation of cell population data on the UniCel DxH 800 Coulter Cellular Analysis system as a screening for viral infection in children

Introduction: The utility of WBC cell population data (CPD) for the differential diagnosis of viral infection from normal control, bacterial infection, and tuberculosis in children was investigated. Methods: A data set of 602 total whole‐blood samples were analyzed on the DxH 800 System for complete blood cell count (CBC) with leukocyte differential from children with the following sample breakdown: 77 confirmed diagnoses of viral infections (Epstein–Barr virus; 30, influenza A; 19, rota virus; 11, other viruses;17), 54 normal control, 71 bacterial infection, 17 TB patients, and 383 with various diseases. The mean (MN) and standard deviation (SD) of the volume (V), conductivity (C), five light‐scatter measurements, and 14 calculated parameters were obtained for the leukocytes. Results: Using a combination of the CBC and CPD parameter values, a decision rule, composed of 21 parameters, for the screening of viral infection in children was developed. Using this decision rule, 74 of 77 (96.1%) viral infections, two of 54 (3.7%) normal samples, one of 17 (5.9%) TB, and six of 71 (8.5%) bacterial infection samples were identified. The sensitivity was 96.1%, and specificity for normal control was 96.3% with an overall specificity of 93.7%. Fifty‐nine samples of 383 samples (15.4%) collected from in‐patient children with various diseases without confirmation of viral infection were included in this decision rule. Conclusion: In conclusion, the implementation of leukocytes CPD parameters can be useful in the detection of viral infection in children.     

Detection of DNA point mutations with DNA mismatch repair enzymes

We have developed a simple and reliable procedure to screengene mutations using DNA mismatch repair (MR) specific mut Yenzyme of Escherichia coli and thymidine DNA glycosylase fromHeLa cells. The mut Y enzyme cleaves A of G/A mismatches inDNA duplex and thymidine glycosylase cleaves T at G/T mismatches.Previously, we showed the determination of G:C – T: Amutations in the N–ras gene in two human tumor sampleswith mut Y G/A MR enzyme. As low as 1–2% mutant DNAs ina sample of mutant and wild-type DNA can be detected with asynthetic DNA to create G/A mispairing for the assay. In thispaper, we simplify the assay, include G/T MR thymidine glycosylasefrom HeLa cells and evaluate the application for screening DNApoint mutations of p53 and ras genes. In this method, DNA fragmentsamplified from normal and mutated genes by polymerase chainreaction (PCR) were mixed and annealed to create DNA mismatchesfor cleavage by mismatch repair enzymes. The cleaved productsand the substrates were separated by gel electrophoresis anddetected by autoradiography. In theory, the enzymes that cutG/A or G/T mispairs will detect the mutations of G:C –A:T, A:T – G:C, G:C – T:A and T:A – G:C. Severalhuman tumor samples were examined for p53 or K-ras mutationswith G/A and G/T mismatch repair enzymes. The reliability ofmutation detection was evaluated by comparing the results withreported mutations or confirmed by DNA sequencing of the samePCR-amplified DNA fragments. Our data showed that, followingmismatch repair enzyme cleavage, all mutated DNA samples yieldedcleaved products with sizes as expected. In addition, our assayis able to characterize the nature of mutation by 5' end-labelingof ³²P on mutant or wild-type DNA fragments. The low background,reliability and the determination of the sites of mutationsas well as the types of DNA base changes indicate the advantagesof the method over other techniques in testing DNA mutants.     

Effects of cyclosporine on glucose metabolism

Cyclosporine may have deleterious effects on glucose metabolism. This study was designed to characterize more precisely cyclosporine-induced alterations in glucose homeostasis in a large animal model with hyperglycemic and euglycemic clamp studies in addition to simple bolus glucose (IVGTT) and insulin (IVITT) tolerance tests. In experiment 1, IVGTTs and hyperglycemic clamp studies were performed in eight ewes before and after 4 weeks of cyclosporine treatment. Studies were repeated 4 weeks after cessation of therapy. In experiment 2, IVITTs and euglycemic clamp studies were performed in seven ewes before and after 4 weeks of cyclosporine treatment. Fasting glucose and insulin levels were not affected by cyclosporine treatment. In experiment 1 cyclosporine did not alter IVGTTs; however, during sustained hyperglycemia, cyclosporine caused a 37% decrease in net glucose disposal (p less than 0.001) and a 39% decrease in plateau plasma insulin levels (p less than 0.05). In experiment 2 cyclosporine had no effect on IVITTs. Plateau insulin values in euglycemic clamp studies were lowered by 27% (p less than 0.05) after cyclosporine treatment. In addition, the metabolic clearance rate of insulin was increased by 25% (p less than 0.05), and the steady-state insulin clearance rate was increased by 16% (p less than 0.003). Measurements of insulin sensitivity were unchanged by cyclosporine. These experiments suggest that cyclosporine treatment results in impairment of sustained synthesis and secretion of insulin, increased insulin clearance, and unaltered insulin sensitivity.     

Systematic facial nerve monitoring in middle ear and mastoid surgeries: “Surgical dehiscence” and “electrical dehiscence”

OBJECTIVES: To evaluate and systemize intraoperative facial nerve monitoring (IOFNM) in middle ear and mastoid surgeries. STUDY DESIGN AND SETTING: A prospective study. METHODS: IOFNM was performed in 100 patients undergoing middle ear and mastoid surgeries. We checked "surgical dehiscence" under microscopes, and also estimated the minimal threshold of electric current needed to change the electromyography of facial muscles using Nerve Integrity Monitor (NIM)-2 (Xomed, Minneapolis, MN, USA). RESULTS: Forty-three percent of cases showed "surgical dehiscence" and responded to electric stimulation of 0.7 mA or less. "Electrical dehiscence" (相似文献   

Quantitative comparisons of antibody-binding sites of platelet glycoprotein IIb/IIIa in aplastic anemia and idiopathic thrombocytopenic purpura

The numbers of antibody-binding sites of platelet glycoprotein (GP) IIb/IIIa on circulating platelets were analyzed using 4 kinds of antibodies in 34 aplastic anemia (AA) patients, 20 idiopathic thrombocytopenic purpura (ITP) patients, and 14 normal controls. The numbers of antibody-binding sites of CD41, CD41a, CD41b, and CD61 on platelets of the AA patients were less than in the normal controls (p <0.001). In the ITP patients, the numbers of sites for CD41 and CD41a were less than in normal controls (p <0.05). There were significant positive correlations between CD41 and CD41a, CD41b, and CD61 in the 3 groups. There were significant negative correlations between CD41 and CD41b and between CD41a and CD41b in the normal controls, but not in the AA or ITP patients. In summary, the numbers of the 4 antibody-binding sites of GPIIb/IIIa on platelets of AA and ITP patients are different from those in normal controls. Measurements of the antibody-binding sites of GPIIb/IIIa are not necessary for the differential diagnosis of AA and ITP. However, the differences in correlations between the numbers of epitopes in AA and ITP patients suggest that the epitopes of GPIIb/IIIa are altered in these diseases.     

Comparative study of immunofluorescent antinuclear antibody test and line immunoassay detecting 15 specific autoantibodies in patients with systemic rheumatic disease

Based on the currently proposed algorithms, antibodies specificities (sp-ANAs) are identified mainly in samples positive for fluorescent antinuclear antibodies (FANA) screening tests. The purpose of the present study was to compare diagnostic performances of FANA and line immune assay (LIA) detecting 15 sp-ANAs in patients with systemic rheumatic diseases (SRD). In 948 sera from the patients with SRD (n = 590) and non-SRD (n = 358), we evaluated the fluorescent patterns and intensities in the FANA test, and compared the FANA results with sp-ANAs against nRNP, Sm, SS-A, Ro52, SS-B, Scl-70, PM/Scl, Jo-1, CENP B, PCNA, dsDNA, nucleosome, histone, ribosomal-P, and M2. The sensitivity and specificity was 75.9% and 52.5% of FANA test and 62.0% and 84.4% of sp-ANAs test for SRD detection. The overall agreement between FANA and sp-ANAs results was 69.2% (Kappa coefficient; 0.404). According to the clinical diagnosis, the levels of agreement varied from 33.3% to 83.1%. The positive predictive values of each FANA pattern for the detection of sp-ANAs were less than 50% except for the discrete speckled pattern (91.7%). The 1:100 intensity of FANA as well as the monoreactivity of LIA, anti-SSA(-)/anti-Ro52(+), or FANA(-)/sp-ANAs(+) was associated with non-SRD. Antibodies against ribosomal-P or PCNA were specific for systemic lupus eryhthematosus. This study highlights the need for careful interpretation of FANA test results to assess sp-ANAs and the application of sp-ANAs tests including less-common autoantibodies. In patients with clinical suspicion of SRD, screening with both FANA and sp-ANAs tests could improve diagnostic efficiency.     

Metabolic studies of HgCl2-induced acute renal failure in the rat

HgCl₂ was used to produce acute renal failure (ARF) in rats. It caused significant decreases (P < 0.001) in ATP, ADP, and total adenine nucleotide levels to 50% of controls at 48 hr with no change in AMP levels. Lactate increased to threefold control levels at 6 hr and remained significantly elevated (P < 0.001) at 48 hr. At 24 hr, there was widespread necrosis in the pars convoluta and pars recta. At 48 hr, necrosis was accompanied by some regeneration, and creatinine levels exceeded 5 mg/dl. Dithiothreitol (30.8 mg/kg, ip, 30 min after HgCl₂) partially ameliorated the functional lesion (creatinine levels ?3.6 mg/dl) and resulted in reduced necrosis in the pars convoluta, but did not significantly alter the metabolic pattern. The results indicate an early metabolic disturbance and are consistent with the idea that HgCl₂ exerts a direct action on the enzymes of the mitochondrial electron transport chain. The failure of dithiothreitol to alter the metabolic pattern of HgCl₂-induced ARF indicated that its protective effect was probably not mediated directly through the maintenance of adenine nucleotide levels.     

Metabolic studies of postischemic acute renal failure in the rat

Postischemic acute renal failure was induced by 1 hr of clamping of the renal vasculature. Adenine nucleotide (ATP, ADP, AMP) and lactate (Lac) levels were measured after 0, 0.25, 1, 6, 24, and 48 hr of reflow to determine the time necessary for recovery to control levels. After 1 hr of ischemia with no reflow, [ATP] was 18% and [Lac] was 10-fold control levels. Control levels were restored after 24 hr of reflow. Variable ischemic times (5, 15, 30, 60, 90, and 120 min) followed by (1) no reflow or (2) 24 hr of reflow were also studied. [ATP] decreased to 25 and 13% of controls after 5 and 120 min of ischemia, respectively, and [Lac] increased to 5- and 13-fold controls after 5 and 120 min. Five to ninety minutes of ischemia followed by 24 hr of reflow resulted in a trend toward restoration of ATP and Lac levels; whereas, 120 min of ischemia followed by 24 hr of reflow resulted in death. The results indicate that: (1) In vivo ischemia results in a drastic and rapid shift in the ATP-ADP-AMP equilibrium; (2) the absolute concentration of ATP is not a reliable criterion of cell viability, but the ability to resynthesize ATP may be determinant in the reversibility of the lesion; (3) 1 hr of ischemia is reversible with respect to restoration of [ATP] and [Lac], but 24 hr of reflow are needed for restoration; and (4) ischemia for 90 min results in a metabolic derangement which is partially reversible in that metabolite levels are partially restored after 24 hr of reflow. However, 90 min of vascular clamping is not functionally reversible since the majority of animals exhibit severe azotemia and do not survive.     

Management for the Children with Otitis Media with Effusion in the Tertiary Hospital

Objectives

Recently, new evidence-based recommendations have been introduced for diagnosing and managing otitis media with effusion (OME) in children. However, there are some difficulties to follow the general guidelines in the tertiary hospitals. The purpose is to evaluate the efficiency of antibiotics or antihistamines for treatment of children with OME in the tertiary hospital with a randomized prospective clinical study.

Methods

Eighty-four children with OME who had been diagnosed in the tertiary hospital were randomized to receive 5 different medications for 2 weeks. We prescribed antibiotics (amoxicillin-clavulanate syrup) in Group I (n=16), antibiotics/steroids (prednisolone) in Group II (n=18), antibiotics/antihistamines (ebastine) in Group III (n=15), antibiotics/steroids/antihistamines in Group IV (n=17), and mucolytics (ivy leaf extract) in Group V (n=17) for control. We followed-up children every 2 weeks and evaluated the state of OME at 3 months.

Results

Thirty six (42.9%) of 84 children were resolved within average 6.9 weeks after the treatments. Thirty-six (42.9%) were treated with ventilation tube insertion and 12 patients (14.3%) were observed. There was no difference in the resolution rates of OME among the five different protocols (P>0.05). There was no difference in the resolution rates among groups who used steroids, antihistamines, steroids and antihistamines, or other medications to manage 42 children with allergies (P>0.05).

Conclusion

In the tertiary hospital, the cure rate of children with OME was not as high as well-known, and antibiotics or anti-allergic medications were not more effective than control. We may, therefore, need any other guidelines which are different from the previous evidence-based recommendations, including early operation in the tertiary hospitals.