Establishing the utility of multi-platform liquid biopsy by integrating the CSF methylome and proteome in CNS tumours.

BACKGROUND: Liquid biopsy represents a major development in cancer research, with significant translational potential. Similarly, it is increasingly recognized that multi-omic molecular approaches are a powerful avenue through which to understand complex and heterogeneous disease biology. We hypothesize that merging these two promising frontiers of cancer research will improve the discriminatory capacity of current models and allow for improved clinical utility. METHODS: We have compiled a cohort of patients with glioblastoma, brain metastasis, and primary central nervous system lymphoma. Cell-free methylated DNA immunoprecipitation (cfMeDIP) and shotgun proteomic profiling was obtained from the cerebrospinal fluid (CSF) of each patient and used to build tumour-specific classifiers. RESULTS: We show that the DNA methylation and protein profiles of cerebrospinal fluid can be integrated to fully discriminate lymphoma from its diagnostic counterparts with perfect AUC of 1 (95% confidence interval 1-1) and 100% specificity, significantly outperforming single-platform classifiers. CONCLUSIONS: We present the most specific and accurate CNS lymphoma classifier to date and demonstrates the synergistic capability of multi-platform liquid biopsies. This has far-reaching translational utility for patients with newly diagnosed intra-axial brain tumours.

[Relationship between macular hole cavity cross-sectional area and retinal blood flow density and its impact on retinal function in idiopathic macular holes].

Objective: To investigate the correlation between the cross-sectional area of the macular hole (MH) cavity and the blood flow density in different retinal layers, as well as the impact of cavity size on preoperative and postoperative retinal function in patients with idiopathic macular holes (IMH). Methods: A retrospective cohort study was conducted. Clinical data were collected from 18 patients (18 eyes) diagnosed with IMH who underwent vitrectomy combined with internal limiting membrane peeling at Shanxi Eye Hospital affiliated to Shanxi Medical University from August 2019 to December 2021. Visual acuity, best-corrected visual acuity (BCVA), spectral-domain optical coherence tomography (SD-OCT), optical coherence tomography angiography (OCTA), and multifocal electroretinography (mfERG) were performed preoperatively and at 1, 3, and 6 months postoperatively. Blood flow density data were collected for the overall retina, macular center fovea, and parafoveal regions, and the cavity cross-sectional area was measured using ImageJ software. Ten healthy subjects without retinal abnormalities who underwent mfERG at Shanxi Eye Hospital affiliated to Shanxi Medical University during the same period were included as a control group. One eye was randomly selected for each subject, with 5 left eyes and 5 right eyes. Statistical analysis was performed using Pearson correlation analysis, independent samples t-test, and repeated measures analysis of variance. Results: All 18 IMH patients had unilateral disease, including 3 males and 15 females, with an average age of (64.22±4.33) years and a duration of illness of 75 (38, 120) days. In the control group, consisting of 10 subjects, 4 were male and 6 were female, with an average age of (63.67±6.96) years. Preoperatively, all 18 affected eyes exhibited a macular hole cavity, which closed and disappeared one month postoperatively. The total cavity cross-sectional area was (4.84±2.28) mm2, with an average area of (0.27±0.13) mm2. The cavity cross-sectional area was positively correlated with the MH height (r=0.82, P<0.001), ellipsoid zone disruption diameter (r=0.74, P<0.001), and preoperative BCVA (r=0.62, P=0.006). The cavity cross-sectional area was positively correlated with the blood flow density of the superficial macular center fovea (r=0.47, P=0.049) and negatively correlated with the blood flow density of the deep retina (r=-0.50, P=0.033) and deep parafoveal blood flow density (r=-0.65, P=0.003). Compared to healthy eyes in the control group, IMH eyes showed decreased amplitudes of P1 and N1 waves in mfERG, prolonged P1 wave latency except at ring 1 (2.18° outward from the foveal center), and prolonged N1 wave latency except at ring 2 (7.46° outward from the foveal center) and 5 (29.75° outward from the foveal center) (all P<0.05). The cavity cross-sectional area was only related to the preoperative latency of the N1 wave at ring 2 (r=0.64, P=0.004) and had an effect on the changes in the preoperative and postoperative latency of P1 and N1 waves at ring 1 and 5 (F=4.94, 5.96; P=0.042, 0.027). Time changes had no effect on the preoperative and postoperative amplitudes and latencies of mfERG P1 and N1 waves (all P>0.05), but the interaction between cavity cross-sectional area and time had statistical significance for the amplitudes of P1 at ring 1 and N1 at ring 2(F=6.89, 3.76; P=0.003, 0.035). Conclusions: In patients with IMH, a larger macular hole cavity cross-sectional area is associated with lower blood flow density, particularly in the deep parafoveal region, poorer visual acuity, and decreased retinal function at 7.46° outward from the foveal center.

UPDATED UNDERSTANDING OF THE MOLECULAR TARGETS OF RADIOIODINE IN DIFFERENTIATED THYROID CANCER.

Radioactive iodine (RAI) therapy is a mainstay adjuvant treatment for thyroid cancer. Administration of RAI therapy after total or near-total thyroidectomy has shown a survival advantage in numerous properly selected patients. However, the role of RAI therapy after reoperation for persistent or recurrent differentiated thyroid carcinomas (DTCs) is unclear. One reason may be the possible downregulation of the I- transport system after primary surgery. RAI is transported by the sodium iodide symporter (NIS), PENDRIN, anoctamin 1 (ANO1) and cystic fibrosis transmembrane conductance regulator (CFTR) and emits ß particles that destroy follicular cells. The identification of pathways of iodide (I-) transport has allowed use of the transport system to render tumours susceptible to RAI treatment via gene therapy. This review focuses on the effect of RAI therapy in follicular cell-derived thyroid cancers and offers potential novel targets that enable improved radioiodine uptake and thus an improved prognosis of thyroid cancer.

Quality of life after endoscopic skull base surgery with a nasoseptal flap: a systematic review.

BACKGROUND: The past two decades have seen a widespread adoption of endoscopic skull base surgery with the use of nasoseptal flaps. Attention has been diverted in recent times to evaluate the quality of life of these patients. OBJECTIVE: This systematic review aimed to evaluate the available literature to compare the quality of life after endoscopic skull base surgery with or without nasoseptal flaps. METHODS: This systemic review was conducted using PubMed, Embase and Cochrane Library databases for literature published after 2009. RESULTS: The majority of studies concluded that there was no statistically significant difference in the quality of life associated with the use of nasoseptal flaps. Post-operatively, more extensive surgery, peri-operative radiotherapy, smoking and younger age were associated with poorer quality of life. CONCLUSION: While the use of nasoseptal flaps can have negative effects on patients' quality of life in terms of sinonasal symptoms, this systematic review found no difference in quality of life associated with the use or non-use of a nasoseptal flap.

A prospective observational prevalence study of elevated HbA1c among elective surgical patients.

Type 2 Diabetes Mellitus (DM) is a chronic disease with high prevalence worldwide. Using glycated haemoglobin (HbA1c) as a surrogate for potential pre-DM and DM conditions, our primary objective was to determine the HbA1c epidemiology in non-cardiac elective surgical patients in Singapore. Our secondary aim was to identify risk factors associated with elevated HbA1c. We conducted a prospective, observational single-centre study in adult patients. HbA1c screening was performed. Patient demographics and comorbidities were recorded. Patients were divided into those with HbA1C ≤ 6.0% and HbA1C ≥ 6.1%. Regression analyses were performed to identify associated factors. Subgroup analysis was performed comparing patients with HbA1C ≥ 6.1% and HbA1C ≥ 8.0%. Of the 875 patients recruited, 182 (20.8%) had HbA1c ≥ 6.1%, of which 32 (3.7%) had HbA1c ≥ 8%. HbA1C ≥ 6.1% was associated with Indian ethnicity [1.07 (1.01-1.13), p = 0.023], BMI > 27.5 [1.07 (1.02-1.11), p = 0.002], higher preoperative random serum glucose [1.03 (1.02-1.04), p < 0.001], pre-existing diagnosis of DM [1.85 (1.75-1.96), p < 0.001] and prediabetes [1.44 (1.24-1.67), p < 0.001], and peripheral vascular disease [1.30 (1.10-1.54), p = 0.002]. HbA1c ≥ 8% had an additional association with age > 60 years [0.96 (0.93-0.99), p = 0.017]. The prevalence of elevated HbA1c is high among the surgical population. Targeted preoperative HbA1c screening for at-risk elective surgical patients reduces cost, allowing focused use of healthcare resources.

Accelerometer-measured physical activity and sedentary behavior in Chinese children and adolescents: a systematic review and meta-analysis.

OBJECTIVE: To synthesize evidence on accelerometer-measured moderate-to-vigorous physical activity (MVPA) and sedentary behavior (SB) levels of Chinese children and adolescents. STUDY DESIGN: This is both a systematic review and meta-analysis study. METHODS: Online databases were searched for studies published from January 2009 up to February 2019. These studies reported accelerometer-measured daily minutes of MVPA and/or SB among Chinese children and adolescents. Random-effects meta-analysis was used to separately pool the time spent in MVPA and SB. RESULTS: Of 4754 records, 20 studies were included in the meta-analysis. Sample sizes ranged from 96 to 2163. The meta-analysis showed that Chinese children and adolescents spent 41.11 min/day in MVPA and 529.83 min/day in SB averagely. Boys spent more time in MVPA compared with girls (P = 0.01). Children accumulated more MVPA time than adolescents (P = 0.05), and children spent less time in SB than adolescents (P = 0.05). Unlike weekdays, SB was lower on weekends (P = 0.02). There were significant differences in children and adolescents' MVPA time in regions (P < 0.001). CONCLUSIONS: MVPA level in Chinese children and adolescents is well below international recommendations, and their SB level is very high.

Clinical and pathologic predictors of central lymph node metastasis in papillary thyroid microcarcinoma: a retrospective cohort study.

OBJECTIVE: To identify the clinical and pathological predictors of central lymph node metastasis (CLNM) in patients with clinically lymph node-negative papillary thyroid microcarcinoma (PTMC). MATERIALS AND METHODS: Data pertaining to 541 clinically lymph node-negative PTMC patients who underwent thyroid surgery at the Shanghai General Hospital between January 2010 and December 2013 were retrospectively analyzed. According to histopathological evidence of central lymph node involvement, patients were divided into central lymph node metastasis (CLNM)-positive and CLNM-negative groups; risk factors for CLNM were identified statistically. RESULTS: LNM was found in 148 (27.4%) patients. Gender (P = 0.002), age (P < 0.001), tumor size (P < 0.001), multifocality (P < 0.001), and extrathyroidal extension (P < 0.001) were significantly different between CLNM-positive and CLNM-negative groups. On multivariate analyses, male sex (odds ratio [OR] = 2.656), age <45 years (OR = 4.184), tumor size >0.575 cm (OR = 2.105), gross extrathyroidal extension (OR = 14.605) and multifocality (OR = 2.084) were independent risk factors for CLNM. Among patients who did not have any of these five risk factors, only 3.9% were found to have CLNM. CONCLUSIONS: A relatively high prevalence of CLNM was observed in patients with clinically lymph node-negative PTMC. CLNM was associated with male sex, younger age, larger tumor size, extrathyroidal extension and multifocal PTMC.

Source Identification of Human Biological Materials and Its Prospect in Forensic Science.

Source identification of human biological materials in crime scene plays an important role in reconstructing the crime process. Searching specific genetic markers to identify the source of different human biological materials is the emphasis and difficulty of the research work of legal medical experts in recent years. This paper reviews the genetic markers which are used for identifying the source of human biological materials and studied widely, such as DNA methylation, mRNA, microRNA, microflora and protein, etc. By comparing the principles and methods of source identification of human biological materials using different kinds of genetic markers, different source of human biological material owns suitable marker types and can be identified by detecting single genetic marker or combined multiple genetic markers. Though there is no uniform standard and method for identifying the source of human biological materials in forensic laboratories at present, the research and development of a series of mature and reliable methods for distinguishing different human biological materials play the role as forensic evidence which will be the future development direction.

Variation in composition and relative content of accumulated photopigments in a newly isolated Rhodobacter capsulatus strain XJ-1 in response to arsenic.

This study aimed to isolate and characterize a new arsenic (As)-tolerant bacterial strain (XJ-1) from the Halosol soil, to evaluate its As tolerance, and to examine the variation in composition and relative content of accumulated photosynthetic pigments in response to As. The experiments were performed with high-performance liquid chromatography (HPLC), inductively-coupled plasma mass spectrometry (ICP-MS), liquid chromatography/mass spectrometry (LC/MS), thin-layer chromatography (TLC) and grayscale intensity image analysis using Gel-Pro analyzer software. Strain XJ-1 was identified as Rhodobacter (R.) capsulatus based on 16S rRNA gene sequencing and physiological characteristics. Strain XJ-1 was able to grow when exposed to arsenite [As(III)] and arsenate [As(V)] under anaerobic-light conditions. The median effective concentrations (EC50) of As(III) and As(V) were 0.61 mM and 2.03 mM, respectively. Strain XJ-1 could reduce As(V) to As(III), but As(III) could not be transformed back to As(V) or other organic As compounds. Accumulation of bacteriochlorophylls and carotenoids in strain XJ-1 varied in the presence of 0.2-1.2 mM As(III) and 0-2.5 mM As(V). As exposure resulted in pronounced variation in compositions and contents of photosynthetic pigments, especially hydroxyspheroidene, bacteriophaeophytin, the ratio of tetrahydrogeranylgeranyl to phytylated BChl a, and the ratio of spheroidene to spheroidenone. This research highlights the adaptative response of R. capsulatus strain XJ-1 photosystems to environmental As, and demonstrates the potential of utilizing the sensitivity of its photosynthetic pigments to As(III) and As(V) for the biodetection of As in the environment.

Characterization of the capsid protein VP2 gene of a virulent strain NE/09 of porcine parvovirus isolated in China.

A virulent strain NE/09 of porcine parvovirus (PPV) was isolated from mummified swine fetus samples in China and its capsid protein VP2 gene was analyzed. The data showed that the VP2 gene encompassed an open reading frame of 1713 nucleotides with a composition of 684 adenine (39.9%), 387 cytosine (22.6%), 268 guanine (15.7%), and 374 thymine (21.8%). The VP2 gene encoded a 570aa-long protein which could be recognized by anti-PPV-VP2 monoclonal antibody 3C9. Sequence analysis revealed that the VP2 gene of PPV-NE/09 had close sequence similarity with the VP2 gene of other PPV strains. However, a 27 nucleotide region that encodes a glycine-rich domain at the N-terminal region of VP2 was deleted in the PPV-NE/09 strain. The PPV-NE/09 virions were purified by 1.39 g/ml CsCl density centrifugation and subjected to SDS-PAGE analysis. The results showed that only two major polypeptides VP1 and VP2 with molecular weights of 83 and 64kDa, respectively were observed, whereas VP3 with a molecular weight of 60 kDa was not present, in contrast to the PPV-NADL-2 strain. According to the phylogenetic relationship of the VP2 gene of PPV-NE/09, the isolate was a new mutant strain of PPV prevailing in China.

Xenobiotic transporters of the human organic anion transporting polypeptides (OATP) family.

1. The organic anion transporting polypeptides (humans OATP; other species Oatp) belong to the SLCO gene superfamily of transporters and are twelve transmembrane domain glycoproteins expressed in various epithelial cells. Some OATPs/Oatps are expressed in a single organ, while others are expressed ubiquitously. 2. The functionally characterized members mediate sodium-independent transport of a variety of structurally independent, mainly amphipathic organic compounds, including bile salts, hormones and their conjugates, toxins, and various drugs. 3. This review summarizes the general features and the substrates of the eleven human OATPs. Furthermore, it reviews what is known about the mechanism of their multispecificity, their predicted structure, their role in drug-food interactions, and their role in cancer. 4. Finally, some open questions are raised that need to be addressed to advance OATP research in the near future.

Cyclosporin A pre-incubation attenuates hypoxia/reoxygenation-induced apoptosis in mesenchymal stem cells.

Although mesenchymal stem cells (MSCs) are being tested for cardiac repair, the majority of transplanted cells undergo apoptosis in the ischaemic heart because of the effects of ischaemia/reperfusion, poor blood supply and other pro-apoptotic factors. Several experimental and clinical studies have suggested that cyclosporin A (CsA) treatment reduces apoptosis in human endothelial cells and neurocytes. However, the effect of CsA on the apoptosis in MSCs is still unclear. In this study, we investigated whether CsA could inhibit hypoxia/ reoxygenation (H/R)-induced apoptosis in MSCs. MSCs pre-incubated with or without CsA were subjected to 6 h of hypoxia followed by 12 h of reoxygenation. Our data showed that pre-incubation with 0.5-5 microM CsA dose-dependently protected the MSCs from H/R injury, as evidenced by decreased apoptosis and increased cell viability. CsA inhibited the H/R-induced translocation of cytochrome c, increased bcl-2 expression and restored mitochondrial membrane potential. CsA also increased the expression of p-BAD. We propose that preincubation MSCs with CsA inhibits MSC apoptosis through the mitochondrial and BAD pathway.

Histone deacetylase (HDAC) inhibitor activation of p21WAF1 involves changes in promoter-associated proteins, including HDAC1.

Histone deacetylase (HDAC) inhibitors (HDACi) cause cancer cell growth arrest and/or apoptosis in vivo and in vitro. The HDACi suberoylanilide hydroxamic acid (SAHA) is in phase I/II clinical trials showing significant anticancer activity. Despite wide distribution of HDACs in chromatin, SAHA alters the expression of few genes in transformed cells. p21(WAF1) is one of the most commonly induced. SAHA does not alter the expression of p27(KIPI), an actively transcribed gene, or globin, a silent gene, in ARP-1 cells. Here we studied SAHA-induced changes in the p21(WAF1) promoter of ARP-1 cells to better understand the mechanism of HDACi gene activation. Within 1 h, SAHA caused modifications in acetylation and methylation of core histones and increased DNase I sensitivity and restriction enzyme accessibility in the p21(WAF1) promoter. These changes did not occur in the p27(KIPI) or epsilon-globin gene-related histones. The HDACi caused a marked decrease in HDAC1 and Myc and an increase in RNA polymerase II in proteins bound to the p21(WAF1) promoter. Thus, this study identifies effects of SAHA on p21(WAF1)-associated proteins that explain, at least in part, the selective effect of HDACi in altering gene expression.

Quantitative design of optimal analgesic combination of acetaminophen, caffeine, and butalbital.

AIM: To quantitatively seek an optimal analgesic combination of acetaminophen (Ace), butalbital (Bul), and caffeine (Caf), and to characterize the pharmacodynamics of interaction among the three drugs. METHODS: The models of acute inflammatory pain in carrageenin-injected rats were applied to measure the vocalization threshold to paw pressure. Six groups with different ratios and doses were set to seek an optimal combination of Ace, Caf, and Bul, analyzed by the weighted modification method. Based on the ratio and doses in the optimal combination, four continuous doses were set to analyze the interactions of therapeutic effects by the reflection method. The interaction of the acute toxicity was evaluated by the parameter method. RESULTS: According to the degree of importance to the combined analgesic effect, Ace > Caf > Bul; Ace showed a significant dose-response relationship, whereas in Caf and Bul, this relationship was not apparent. A new combination was obtained by the theoretical analysis and confirmed further by experimentation. Namely, at a ratio of 8.6:1:1.5 Ace + Caf + Bul (240 + 28 + 42 mg/kg, ig) was an optimal combination. Both Caf and Bul had a synergism to Ace, but Caf was a stronger synergist in contrast to Bul. Such synergism increased the therapeutic effects in the range of Ace 153.6 - 300 mg/kg combined with Caf 17.9 - 35 mg/kg and Bul 26.8 -5 2.5 mg/kg (8.6:1:1.5). However, the dose of Ace + Caf + Bul at 300 + 35 + 52.5 mg/kg resulted in sedation in rats. The peak latency was approximately 1 h for all four continuous doses, but the peak amplitude was dose-related, and the duration of the therapeutic effect was less than 2 h. The acute toxicity of the three drugs in combination remained the same. CONCLUSION: Ace + Caf + Bul at a dose of (240 + 28 + 42) mg/kg (ig) results in an optimal combination. The therapeutic window of the combination is located in the range of Ace (153.6 - 240 mg/kg)+Caf (17.9 - 28 mg/kg)+Bul (26.8 - 42 mg/kg) (8.6:1:1.5). Caf has a stronger synergism with Ace than Bul.

Acetylation of a specific promoter nucleosome accompanies activation of the epsilon-globin gene by beta-globin locus control region HS2.

On stably replicating episomes, transcriptional activation of the epsilon-globin promoter by the beta-globin locus control region HS2 enhancer is correlated with an increase in nuclease sensitivity which is limited to the TATA-proximal nucleosome (N1). To elucidate what underlies this increase in nuclease sensitivity and the link between chromatin modification and gene expression, we examined the nucleoprotein composition and histone acetylation status of transcriptionally active and inactive promoters. Micrococcal nuclease digestion of active promoters in nuclei released few nucleosome-like nucleoprotein complexes containing N1 sequences in comparison to results with inactive promoters. We also observed that N1 DNA fragments from active promoters are of a subnucleosomal length. Nevertheless, chromatin immunoprecipitation experiments indicate that histones H3 and H4 are present on N1 sequences from active promoters, with H3 being dramatically hyperacetylated compared with that from inactive promoters and vector sequences. Strikingly, H3 in the adjacent upstream nucleosome (N2) does not appear to be differentially acetylated in active and inactive promoters, indicating that the nucleosome modification of the promoter that accompanies transactivation by HS2 is highly directed and specific. However, global acetylation of histones in vivo by trichostatin A did not activate transcription in the absence of HS2, suggesting that HS2 contributes additional activities necessary for transactivation. N1 sequences from active promoters also contain reduced levels of linker histone H1. The detection of a protected subnucleosomal sized N1 DNA fragment and the recovery of N1 DNA sequences in immunoprecipitations using anti-acetylated H3 and H4 antibodies argue that N1 is present, but in an altered conformation, in the active promoters.

Function of GATA transcription factors in hydroxyurea-induced HEL cells.

HEL cells, a human erythroleukemia cell line, mainly express the fetal (gamma) globin gene and trace amount of the embryonic (epsilon) globin gene, but not adult (beta) globin gene. Here we show that hydroxyurea (HU) can induce HEL cells to express adult (beta) globin gene and lead these cells to terminal differentiation. Results showed in Gel mobility shift assays that GATA factors could specifically bind to the regulatory elements of human beta-globin gene, including the proximal regulatory element (the beta-promoter) and the distal regulatory elements (the DNase I hypersensitive sites in the LCR, HS2-HS4 core sequences). However, the DNA binding patterns of GATA factors were quite different between HU-induced and uninduced HEL cells. Western-blot analysis of nuclear extracts from both the uninduced and HU-induced HEL cells revealed that the level of GATA-2 transcription factor decreased, whereas the level of GATA-1 transcription factor increased following the time of hydroxyurea induction. Furthermore, using RT-PCR analysis the expression of human beta-globin gene in HU-induced HEL cells could be blocked again when HEL cells were incubated in the presence of antisense oligonucleotides for hGATA-1, suggesting that the upregulation of hGATA-1 transcription factor might be critical for the expression of human beta-globin gene in HU-induced HEL cells.

Esterification of polyunsaturated fatty acids by various forms of immobilized lipase from Rhizomucor miehei.

Ethyl esterification specificity of a lipase from Rhizomucor miehei for polyunsaturated fatty acids (PUFA) was compared at 1 and 100 mM to study molecular recognition of PUFA. The chemical shift of methylene adjacent to carboxyl groups in the nuclear magnetic resonance spectrum of docosahexaenoic acid (DHA) in ethanol moved to a lower magnetic field as the concentration of DHA increased, suggesting that the degree of dissociation of DHA decreased. Specificity constants or apparent second-order rate constants (Vmax/Km or catalytic power) for 1 mM esterification by immobilized lipases were higher than the native lipase. Immobilized hydrophobic carrier of low mass transfer resistance for the esterification substrate may improve maximal velocity and affinity for the substrate. Higher specificity constants for 1 mM substrates were observed using immobilized lipases fixed on an anion exchange resin with glutaraldehyde and on a cation exchange carrier with carbodiimide. Activity yields measured with 1 mM PUFA substrate were high. For the substrates at a concentration of 100 mM, higher specific constants with these bifunctional reagents were not observed but higher activity yields were found.

STAT1 is involved in signal transduction in the EPO induced HEL cells.

Erythropoietin (EPO) is the major regulator of mammalian erythropoisis, which stimulates the growth and differentiation of hematopoietic cells through interaction with its receptor (EPO-R). Here we use HEL cells (a human erythro-leukemia cell line) as a model to elucidate the pathway of signal transduction in the EPO-induced HEL cells. Our data show that the EPOR (EPO receptor) on the surface of HEL cells interacts with the Janus tyrosine protein kinase (Jak2) to transduce intracellular signals through phosphorylation of cytoplasmic proteins in EPO-treated HEL cells. Both STAT1 and STAT5 in this cell line are tyrosine-phosphorylated and translocated to nucleus following the binding of EPO to HEL cells. Furthermore, the binding of both STAT1 and STAT5 proteins to specific DNA elements (SIE and PIE elements) is revealed in an EPO-dependent manner. Our data demonstrate that the pathway of signal transduction following the binding of EPO to HEL cells is similar to immature erythroid cell from the spleen of mice infected with anemia strain of Friend virus.

Mechano-optical waveguide on-off intensity modulator.

High-extinction on-off modulators are essential for channel selection in integrated-optical sensor arrays. We report a standard SiON-technology-based electrostatically driven integrated mechano-optical waveguide on-off intensity modulator. On-off modulation is achieved by movement of an absorbing element into and out of the evanescent field of the guided mode. An extinction ratio of >37 dB at an actuation voltage of <30 V was achieved in a 6 mm x 4 mm device for a wavelength of 632.8 nm. Full wafer-scale fabrication is made possible by use of chemical mechanical polishing and aligned wafer bonding.

The apoptosis of HEL cells induced by hydroxyurea.

Hydroxyurea has been used to synchronize cultured cells to S-phase and used to treat patients with sickle-cell anemia. Recently, we found that hydroxyurea can induce the apoptosis of HEL (human erythroleukemia) cells. The induced HEL cells showed ultrastructurally chromatin condensation with regular crescents at the nuclear edges and apoptotic bodies. However, the cells of K562, another human erythroleukemia cell line, did not show such morphological changes. Under fluoroscope, the HEL cells after induction often displayed a clear reduction in nuclear diameter and nuclear chromatin cleavage and condensation and the presence of nuclear ring and apoptotic bodies. Analysis with flow cytometry showed that the percentage of apoptotic cells is about 30-40% after HEL cells were induced by hydroxyurea for 3 days. DNA ladder can be observed by electrophoretic analysis.