Plasma Separator for Encapsulating Fluorescent Probe in Erythrocytes

Using a plasma separator, it is possible to wash large volumes of whole blood free of plasma proteins in short periods. The novelty, however, is that the same apparatus can be used to reversibly hemolyze and reseal the red blood cells (RBCs) using hypotonic and hypertonic dialysates, respectively. This technique was assessed using the fluorescent exogenous agent uranin (fluorescein sodium). Results indicate that the encapsulation of the RBCs can be completed in under 2 h. The encapsulation efficiency of the RBCs was approximately 80% with a RBC recovery rate of 75%.     

Gastroesophageal reflux disease (GERD), extraesophageal reflux (EER) and recurrent chronic rhinosinusitis

Chronic polypoid rhinosinusitis (CRS) is a common disease, affecting approximately 16% of the adult population in the US every year. In addition to many well known predisposing factors, an association with reflux disease is hypothesized. Such an association might explain the recurrence of polyposis in the face of improved surgical techniques and postsurgical treatment of CRS. At present it is unclear whether extraesophageal reflux directly injures the sinus mucosa, whether gastroesophageal reflux leads to vagus-mediated neuroinflammatory changes, or whether both mechanisms occur separately or simultaneously. In patients suffering from recurrent CRS (n=20) and healthy volunteers (n=20), ambulatory 24 h two channel pH testing was performed. The number of reflux events, the fraction of the total time during which pH was below 4, and the reflux area index (RAI) were determined in the esophagus as well as in the hypopharynx. Patients with recurrent CRS had significantly more reflux events in the esophagus and the fraction of pH<4 and the RAI were increased up to 10-fold compared to healthy volunteers. In contrast to the esophagus, these differences were not observed in the hypopharynx. Recurrent CRS is often associated with GERD but not with EER. Recurrent disease or prolonged recovery after surgery should raise the suspicion of reflux disease as a possible triggering factor. Because GERD itself cannot be diagnosed by laryngoscopy, and because of the subjectivity of symptoms such as heartburn, the otolaryngologist should consider double-probe pH testing as the diagnostic procedure of choice.     

Mid-infrared spectroscopy on skin using a silver halide fibre probe in vivo

Background/aim: Mid-infrared spectroscopy is a versatile method for in vivo investigation of skin after topical treatment with skin care products.
Methods: FTIR-spectrometer (Bruker Optics) with a flexible silver halide fibre probe (Infrared Fiber Sensors).
Results: Absorbance spectra from 700 to 3000 cm⁻¹ have been recorded to gain information about proteins (amide-I and amide-II vibrations at 1650 and 1550 cm⁻¹), esters (1740 cm⁻¹), carboxylic acid (1710 cm⁻¹), polyalcohols (1050 cm⁻¹) and hydrocarbons (CH _n vibrations at 2800–3000 cm⁻¹).
Conclusions: Using the particular light guide, we were able to measure for the first time the effects of lip care products on lips directly. Furthermore, water binding and glycerol content of the skin could be determined simultaneously, as well as the replenishment of lipids by lipid-enriched bath oil.     

改进微波幅射器探头治疗宫颈糜烂疗效观察

目的：探讨改进的微波辐射器探头治疗宫颈糜烂的疗效。方法：将微波辐射器探头由双直线形、双针形、单针形等点线状改进成为“单舌形”和“双舌形”两种弧形平面探头，用于治疗宫颈糜烂。分析1999-2002年450例宫颈糜烂治疗资料，其中300例经微波治疗(A组)，150例经C02—激光治疗(B组)，随访1—3个月。结果：两组治疗后3个月内A组治愈率明显高于B组(P＜0．005)，总有效率A组明显高于B组(P＜0．025)。两组术中、术后不良反应比较：术中宫颈创面出血A组明显低于B组(P＜0．005)，术中腹痛、脱痂期出血量、术后局部感染率方面两组相似。结论：所改进的微波辐射器探头治疗宫颈糜烂治愈率高，操作安全，术后并发症少，医患乐于接受。     

非放射性HBV－DNA探针在乙型肝炎病毒检测中的应用

以地高辛甙元随机引物法标记ＨＢＶ－ＤＮＡ探针，以此探针检测慢性乙型肝炎患者的血清、肝组织，同时以ＥＬＩＳＡ法检测血清ＨＢｅＡｇ、ＨＢｃＡｂ。结果：血清ＮＢｅＡｇ阳性率２７％（１０／３７），血清ＨＢＶ－ＤＮＡ检出率５７．１％（２０／３５），两者有显著性差异。血清ＨＢｃＡｂ阳性率７８．４％（２９／３７），肝组织ＨＢＶ－ＤＮＡ检出率８３．８％（３１／３７），两者无显著性差异。血清与肝组织ＨＢＶ－ＤＮＡ检出率有显著性差异。提示：血清ＨＢＶ－ＤＮＡ检测是较ＨＢｅＡｇ更为准确客观反映血液带毒状况的指标。而准确反映肝脏带毒状况的指标是肝组织ＨＢＶ－ＤＮＡ检测。当ＨＢｅＡｇ阴转，血清ＨＢＶ－ＤＮＡ阴性而肝组织ＨＢＶ－ＤＮＡ阳性时，需注意肝硬化及肝癌的发生。     

Applications of DNA Flow Cytometry and Fluorescence In situ Hybridization Using a Chromosome-specific DNA Probe on Paraffin-embedded Tissue Sections of Primary Malignant Melanomas

We have applied DNA flow cytometric analysis to paraffin-embedded tissue sections of primary malignant melanomas. Conventionally, flow cytometric analysis of paraffin-embedded tissue sections has been done by the method of Hedley et al. We added ultrasound treatment to the method of Hedley et al. and a lower value of coefficient of variation was shown. Furthermore, a new technique, fluorescence in situ hybridization with a chromosome-specific repetitive DNA probe, was used for the analysis of chromosomal numerical aberrations in the same paraffin-embedded tissue sections. The DNA flow cytometric analysis showed that in 8 cases six primary malignant melanomas were of the aneuploid pattern and two cases of lentigo maligna (melamona in situ) were of the diploid pattern. By fluorescence in situ hybridization, the two cases with the diploid pattern had spots/nucleus of 1.28 and 1.12, and those with the aneuploid pattern had spots/nucleus from 2.01 to 2.27. Only one nodular melanoma in an aneuploid case showed spots/nucleus of 1.71. These data indicate that fluorescence in situ hybridization with chromosome-specific repetitive DNA probes can serve as a cytogenetic tool for the analysis of interphase nuclei of solid human tumors and may be useful for the study of tumor cell heterogeneity.     

Orthogonal solid-phase synthesis of a monobiotinylated analog of Neuropeptide Y

Analogs of Neuropeptide Y (NPY) were synthesized with conventional Boc/benzyl protective group strategy. Instead of Asn⁷ in the native scquence, Boc-Lys(Alloc)-OH was incorporated. At the end of the synthesis the Alloc group was selectively removed by palladium-catalyzed hydrostannolysis and biotin coupled to the e-amino group of Lys⁷. After cleavage and characterization with plasma desorption mass spectrometry the N^e-7-biotinyl-[Lys⁷]-NPY and the nonbiotinylated analog [Lys⁷]-NPY were investigated as ligands to the NPY receptor from rat cerebral cortex. Both analogs were found to be high affinity ligands to the NPY receptor and bound with essentially the same affinity as unmodified NPY.     

Hemoglobin constant spring defined by specific oligonucleotide hybridization and hemoglobin D Punjab (beta 121----Gln) in a Batak Indonesian family

A Batak Indonesian from North Sumatra with hemoglobin (Hb) D Punjab (alpha 2 beta 2 121----Gln) and hemoglobin Constant Spring (Hb CoSp) is described. The 24-year-old man did not have clinical symptoms, and his hematological indices were normal. However, he had a persistent slight elevation of fetal hemoglobin level. His mother and his brother were heterozygous for Hb D Punjab; his father had Hb CoSp trait. A sister did not have any abnormal hemoglobin. To show the exact molecular defect leading to the synthesis of Hb CoSp in this family, genomic DNA from the father was analyzed by hybridization with synthetic oligonucleotides. Genomic DNA was digested with Sst I and Hind III producing a 1.05-kb fragment from the 3' end segment of the alpha 2-globin gene, including the termination codon. Two nonadecamers were synthesized to serve as probes: one, entirely homologous to the normal 3' end of alpha 2A-globin gene sequence, including the termination codon TAA, the other different from it by a replacement of the T in the termination codon TAA with C, changing it to CAA, the codon for the amino acid glutamine. DNA from normal controls gave a positive signal with the normal alpha 2TAA oligonucleotide probe but negative with the alpha 2 CAA probe. The father of propositus who had Hb CoSp trait gave a positive signal with the normal alpha 2TAA oligonucleotide probe as well as with the alpha 2CAA oligonucleotide probe, showing him to be heterozygous for the alpha 2CAA-globin gene. This result shows that the Hb CoSp in the Batak family is indeed due to a replacement of T by C in the TAA termination codon of the alpha 2-globin gene changing it to CAA the condon for glutamine. This explains the resulting readthrough of the untranslated sequence of the mRNA.     

DNA探针的高效标记和哺乳动物细胞转化子的检测

本实验采用随机引物法标记探针DNA.探针DNA加热变性成单链,以单链DNA为模板,六聚脱氧核苷酸为引物,在DNA多聚酶I大片段的作用下进行放射性标记.标记探针的比放射性达10~8cpm/μg DNA以上.放射性底物的掺入率为60%.用该方法标记的人高度重复顺序探针,可以检出微微克水平的阳性分子.与人HeLa S_3细胞DNA转化的小鼠LTK~+细胞DNA打点杂交呈阳性,与未经转化的小鼠LTK~-细胞DNA杂交为阴性,证明小鼠转化细胞中存在HeLa细胞DNA,DNA转化是成功的.