Rapid and reliable genotyping of human platelet antigen (HPA)-1, -2, -3, -4, and -5 a/b and Gov a/b by melting curve analysis

BACKGROUND: The probability for occurrence of neonatal alloimmune thrombocytopenic purpura (NAITP) depends largely on the frequency of each individual phenotype in various populations. In caucasians, antibodies to human platelet antigen (HPA)-1a are the major cause of neonatal alloimmune thrombocytopenic purpura, whereas in the Japanese population, antibodies to HPA-4b is most frequently involved in NAITP. Conventional PCR techniques for platelet antigen genotyping rely on sequence-specific primers (SSPs) and detection by gel electrophoresis, a method which is laborious and time consuming. New PCR technology, measuring the match of a hybridization probe with its target and thereby allowing simultaneous detection of both alleles, provides an efficient tool for genotyping of the HPA systems. STUDY DESIGN AND METHODS: A total of 105 healthy blood donors were genotyped for HPA-1, -2, -3, -4, and -5 a/b and Gov a/b with new primers and probes designed for mutation detection by melting curve analysis (using LightCycler technology). Donor DNA was independently genotyped by an allele-specific assay, using SSPs, in a reference laboratory. RESULTS: There was full concordance between the two genotyping methods, and genotype frequencies were comparable with previous studies in caucasians. CONCLUSION: We present rapid and reliable detection systems for HPA-1, -2, -3, -4, and -5 a/b and Gov a/b based on mutation detection of both alleles simultaneously by melting curve analysis. As the Gov system has been reported to have similar frequency of involvement in alloimmune thrombocytopenia as HPA-5, the opportunity for genotyping should aid the diagnosis of such patients.     

Tongue movements in normal 2-, 3-, and 4-year-old children: a continuation study

In this study, tongue movements of normal 2-, 3-, and 4-year-old children were measured and compared. The 4-year-olds were also compared with 4-year-old children in a previous study (Schwartz J: Observation of Tongue and Jaw Movement in Normal Four- and Five-Year-Old Children, master's thesis. Washington University, School of Occupational Therapy, St. Louis, MO, 1982). Measures were taken on 56 children: 17 two-year-olds (8 females, 9 males), 19 three-year-olds (10 females, 9 males), and 20 four-year-olds (10 females, 10 males). Two different tongue positions were quantified: First, the position of the tongue as the food was presented to the child at the moment when the food was 5 cm away from the lips and second, the position of the tongue as the food was swallowed. Younger children held their tongues in a more forward position when the food was presented, although the predominant tongue position was behind the teeth in all three age groups. During swallowing there was also a progression, as age increased, from pursing the lips to puckering at the corners of the mouth. Many children kept their mouth open during swallowing. These results provide further normative data for the assessment of eating-impaired children.     

14-3-3sigma, a p53 regulator, suppresses tumor growth of nasopharyngeal carcinoma

The 14-3-3sigma gene product, up-regulated by p53 in response to DNA damage, is involved in cell-cycle checkpoint control and is a human cancer epithelial marker down-regulated in various tumors. However, its role and function have not been established in nasopharyngeal carcinoma (NPC), a tumor of epithelial origin. Recently, we found that 14-3-3sigma interacts with p53 in response to DNA damage and stabilizes the expression of p53. In addition, we also showed that overexpression of 14-3-3sigma inhibits oncogene-activated tumorigenicity. In the present study, we investigated the tumor-suppressive role of 14-3-3sigma in NPC cells. We found that there is a failure to up-regulate 14-3-3sigma in response to DNA damage in two NPC cell lines that have p53 mutation. We also found that 14-3-3sigma interacted with protein kinase B/Akt and negatively regulated the activity of Akt. Overexpression of 14-3-3sigma inhibited NPC cell growth and blocks DNA synthesis. Overexpression of 14-3-3sigma also led to inhibition of anchorage-independent growth of NPC cells. In addition, we found that 14-3-3sigma sensitized NPC cells to apoptosis induced by the chemotherapeutic agent 2-methoxyestradiol. Overexpression of 14-3-3sigma in both NPC cell lines reduced the tumor volume in nude mice, which could have significance for clinical application. These findings provide an insight into the roles of 14-3-3sigma in NPC and suggest that approaches that modulate 14-3-3sigma activity may be useful in the treatment of NPC.     

Platelet alloantigens HPA-1, -2, -3, -5 and -6b in Finns

SUMMARY. The human platelet alloantigens HPA-1, -2, -3, -5 and -6b in the Finnish population were determined using allele specific restriction analysis (PCR-ASRA) for HPA-1, -2, -3 and -5 and monoclonal antibody immobilized platelet antigen (MAIPA) assay for HPA-1, -3a, -5b and -6b. No discrepancies were observed between the results obtained with the PCR-method and those obtained serologically. The gene frequencies obtained from 200 unrelated Finns were 0.86 and 0.14 for HPA-1a and -1b, 0.91 and 0.09 for HPA-2a and -2b, 0.59 and 0.41 for HPA.3a and -3b and 0.95 and 0.05 for HPA-5a and -5b. The frequency of the HPA-5b allele (10%) is lower in Finns than in Central- or South-European populations (20–30%). The HPA-1, -2 and -3 frequencies did not deviate from those observed in other European populations. The rare HPA-6b antigen was observed in three of 127 individuals from south-eastern Finland (2.4%), which suggests that the frequency of this allele in Finland is higher than previously thought.     

Caspases -2, -3, -6, and -9, but not caspase-1, are activated in sepsis-induced thymocyte apoptosis

Sepsis induces extensive lymphocyte cell death that may contribute to immune depression and morbidity/mortality in the disorder. bcl-2 is a member of a new class of oncogenes that prevents cell death from an array of noxious stimuli. Transgenic mice that overexpress BCL-2 in T lymphocytes are resistant to sepsis-induced T cell apoptosis, and mortality was decreased in sepsis. The purpose of this study was to identify key initiator and executioner "caspases" involved in sepsis-induced lymphocyte apoptosis and to determine if BCL-2 acts prior to caspase activation. Thymi were removed 5-22 h post-cecal ligation and puncture (CLP) or sham surgery. Apoptosis was evaluated in thymocytes by annexin-V FITC labeling and flow cytometry. Caspase-1 activity was determined by western blot analysis of the procaspase protein and p20 subunit of the activated caspase; activities of caspases -2, -6, and -9 were determined by colorimetric assays using specific substrates conjugated to a color reporter molecule. Caspase-3 activity was determined both by western blot and by a fluorogenic assay in which a fluorescent compound was generated. Thymocytes from CLP mice had markedly increased apoptosis and activation of caspases -2, -3, -6, and -9 in comparison with thymocytes of sham-operated mice. Caspase-1 was not activated. BCL-2 prevented sepsis-induced thymocyte apoptosis and inhibited activation of all caspases. We conclude that sepsis causes activation of multiple caspases and that BCL-2 acts upstream as an inhibitor of caspase activation. The pattern of caspase activation suggests a mitochondrial mediated pathway.     

Ferroelectric performance of nylons 6-12, 10-12, 11-12, and 12-12

Nylons have great potential for electrical applications requiring high polarizability and low dielectric loss. Recently, the narrow single hysteresis loop with relaxor ferroelectricity and the double hysteresis loop due to antiferroelectricity have been reported in nylon random copolymers, terpolymers, and common even-numbered nylons. Although several studies of ferroelectric nylons have been reported, even–even-numbered and odd–even-numbered nylons have not been sufficiently explored. Here, the ferroelectricity of spin-coated even–even-numbered and odd–even-numbered nylons was investigated. A series of even–even-numbered nylons, including nylons 6-12, 10-12, and 12-12, and an odd–even-numbered nylon, nylon 11-12, were polymerized with 1,10-dodecanedicarboxylic acid (12) and four aliphatic diamines with various methylene units, 1,6-hexanediamine (6), 1,10-decanediamine (10), 1,11-undecanediamine (11), and 1,12-dodecanediamine (12). The obtained nylon polymers were spin coated and then subjected to melt-quenching or thermal annealing followed by quenching. From the X-ray diffraction and the electrical hysteresis loop data, the correlation between the ferroelectricity and the crystal parameters of crystallinity and crystallite size of the γ crystal phase was investigated. Furthermore, the free volume of the nylon samples was estimated to correlate with the ferroelectricity. Temperature-dependent ferroelectricity was investigated for nylon 10-12. At a high temperature, the nylon samples showed a narrow polarization–electric field hysteresis loop and a rhombus-shaped polarization current–electric field hysteresis loop due to the relaxor ferroelectricity. This behaviour was caused by electrically rotating the nanodomains with weakened hydrogen bonds at higher temperatures.

Nylons have great potential for electrical applications requiring high polarizability and low dielectric loss.     

Antidiarrheal and central nervous system activities of SC-27166 (2-[3 - 5 - methyl - 1, 3, 4 - oxadiazol - 2 - yl) - 3, 3 - diphenylpropyl] - 2 - azabicyclo [2.2.2]octane), a new antidiarrheal agent, resulting from binding to opiate receptor sites of brain and myenteric plexus.

Pharmacological studies were performed to investigate the interaction of SC-27166 (2-[3-(5-methyl-1,3,4--oxadiazol-2-yl)-3,3-diphenylpropyl]-2-azabicyclo[2.2.2]octane), a new antidiarrheal agent, with opiate receptor sites in vitro and in vivo. Morphine, loperamide and SC-27166 inhibited the binding of [3H]naloxone to homogenates of guinea-pig brain and myenteric plexus and the inhibition was diminished in the presence of 100 mM Na+. Unlike that of morphine and [3H]naloxone itself, the binding of loperamide and SC-27166 was complex and Scatchard plots indicated the presence of low and high affinity sites for both compounds. Morphine, loperamide and SC-27166 inhibited the contractions of electrically driven longitudinal muscle from guinea-pig ileum and naloxone antagonized these effects. In the anesthetized dog, i.v. administration of morphine and SC-27166 enhanced the contractile activity of circular muscle in proximal and distal duodenum and distal ileum but duodenal longitudinal muscle was relaxed; these effects were completely reversed by subsequent administration of naloxone. In the rat, p.o. administration of loperamide and SC-27166 inhibited intestinal propulsion at doses considerably lower than were necessary to produce activity in the hot plate test; this specificity of action was not seen with morphine. In the rat, p.o. administration of loperamide and SC-27166 inhibited diarrhea at doses considerably lower than were necessary to produce withdrawal symptoms. The authors concluded that both loperamide and SC-27166 are specific antidiarrheal agents that produce both their central and antidiarrheal effects by binding to opiate receptor sites.     

超短波对脑血管病可溶性自细胞介素α受体及肿瘤坏死因子α、干扰素α的影响

目的 探讨超短波对脑血管病患者可溶性白细胞介素２受体（ＳＩＬ－ ２Ｒ）、肿瘤坏死因子α（ＴＮＦ－α）、干扰素α（ＩＦＮ－α）的影响。方法　　４０例脑血管病患者随机配对分成治疗组、对照组各２０例。两组均接受常规药物治疗,治疗组另外增加胸部超短波治疗作用于胸腺,治疗前后分别测定外周血清ＳＩＬ－２Ｒ、ＴＮＦ－α、ＩＦＮ－α含量变化。结果治疗组治疗后ＳＩＬ－２Ｒ、ＴＮＦ－α均减少（ｔ＝２．９８、３．９２,Ｐ＜０．０１）;ＩＦＮ－α增加（ｔ＝３．８６,Ｐ＜０．０１）。结论　　超短波治疗能提高患者机体的总体免疫平衡状态。