Quantification of bone marrow interstitial pH and calcium concentration by intravital ratiometric imaging.

The fate of hematopoietic stem cells (HSCs) can be directed by microenvironmental factors including extracellular calcium ion concentration ([Ca2+]e), but the local [Ca2+]e around individual HSCs in vivo remains unknown. Here we develop intravital ratiometric analyses to quantify the absolute pH and [Ca2+]e in the mouse calvarial bone marrow, taking into account the pH sensitivity of the calcium probe and the wavelength-dependent optical loss through bone. Unexpectedly, the mean [Ca2+]e in the bone marrow (1.0 ± 0.54 mM) is not significantly different from the blood serum, but the HSCs are found in locations with elevated local [Ca2+]e (1.5 ± 0.57 mM). With aging, a significant increase in [Ca2+]e is found in M-type cavities that exclusively support clonal expansion of activated HSCs. This work thus establishes a tool to investigate [Ca2+]e and pH in the HSC niche with high spatial resolution and can be broadly applied to other tissue types.

Development of the Soft X-ray AGM-AGS RIXS beamline at the Taiwan Photon Source.

We report on the development of a high-resolution and highly efficient beamline for soft X-ray resonant inelastic X-ray scattering (RIXS) located at the Taiwan Photon Source. This beamline adopts an optical design that uses an active grating monochromator (AGM) and an active grating spectrometer (AGS) to implement the energy compensation principle of grating dispersion. Active gratings are utilized to diminish defocus, coma and higher-order aberrations, as well as to decrease the slope errors caused by thermal deformation and optical polishing. The AGS is mounted on a rotatable granite platform to enable momentum-resolved RIXS measurements with scattering angles over a wide range. Several high-precision instruments developed in-house for this beamline are described briefly. The best energy resolution obtained from this AGM-AGS beamline was 12.4 meV at 530 eV, achieving a resolving power of 4.2 × 104, while the bandwidth of the incident soft X-rays was kept at 0.5 eV. To demonstrate the scientific impact of high-resolution RIXS, we present an example of momentum-resolved RIXS measurements on a high-temperature superconducting cuprate, i.e. La2-xSrxCuO4. The measurements reveal the A1g buckling phonons in superconducting cuprates, opening a new opportunity to investigate the coupling between these phonons and charge-density waves.

Combined modalities of resistance in an oxaliplatin-resistant human gastric cancer cell line with enhanced sensitivity to 5-fluorouracil.

To identify mechanisms underlying oxaliplatin resistance, a subline of the human gastric adenocarcinoma TSGH cell line, S3, was made resistant to oxaliplatin by continuous selection against increasing drug concentrations. Compared with the parental TSGH cells, the S3 subline showed 58-fold resistance to oxaliplatin; it also displayed 11-, 2-, and 4.7-fold resistance to cis-diammine-dichloroplatinum (II) (CDDP), copper sulphate, and arsenic trioxide, respectively. Interestingly, S3 cells were fourfold more susceptible to 5-fluorouracil-induced cytotoxicity due to downregulation of thymidylate synthase. Despite elevated glutathione levels in S3 cells, there was no alteration of resistant phenotype to oxaliplatin or CDDP when cells were co-treated with glutathione-depleting agent, l-buthionine-(S,R)-sulphoximine. Cellular CDDP and oxaliplatin accumulation was decreased in S3 cells. In addition, amounts of oxaliplatin- and CDDP-DNA adducts in S3 cells were about 15 and 40% of those seen with TSGH cells, respectively. Western blot analysis showed increased the expression level of copper transporter ATP7A in S3 cells compared with TSGH cells. Partial reversal of the resistance of S3 cells to oxaliplatin and CDDP was observed by treating cell with ATP7A-targeted siRNA oligonucleotides or P-type ATPase-inhibitor sodium orthovanadate. Besides, host reactivation assay revealed enhanced repair of oxaliplatin- or CDDP-damaged DNA in S3 cells compared with TSGH cells. Together, our results show that the mechanism responsible for oxaliplatin and CDDP resistance in S3 cells is the combination of increased DNA repair and overexpression of ATP7A. Downregulation of thymidylate synthase in S3 cells renders them more susceptible to 5-fluorouracil-induced cytotoxicity. These findings could pave ways for future efforts to overcome oxaliplatin resistance.

Design and development of virtual reality based perceptual-motor rehabilitation scenarios.

Virtual reality technology may provide new options for conducting perceptual-motor assessment within simulated 3D environments for persons with a wide range of disabilities. This paper outlines our work developing a series of game-like VR scenarios to assess and rehabilitate eye-hand coordination, range of motion and other relevant perceptual-motor activities. Our efforts have focused on building engaging game-based stereoscopic graphic scenarios that allow clients to participate in perceptual-motor rehabilitation by interacting with 3D stimuli within a full 360-degree space using a head mounted display or by way of a "face-forward" format using 3D projection displays. Exploratory work using multiple video sensors to detect and track 3D body motion, identify body postures and quantify motor performance is also described.

Stroke: who's counting what?

INTRODUCTION: Patients with stroke are often selected for epidemiological reporting and research using ICD-9-CM (ICD-9) diagnostic codes. This study addresses the accuracy of these codes in identifying patients with stroke. METHODS: A sample of 279 patients with new stroke and 392 non-stroke (no evidence of new stroke) patients were identified by medical record review from 11 Veterans Affairs Medical Centers. Administrative records containing ICD-9-CM diagnoses were matched with this sample. Coding sensitivity and specificity were determined using individual ICD-9 codes and two coding algorithms. RESULTS: Significant variation was found in the accuracy of cerebrovascular ICD-9-CM codes in identifying patients diagnosed with stroke. Two coding algorithms were identified with the following performance statistics based on the sampled populations: 1) 91-percent sensitivity, 40-percent specificity; and 2) 54-percent sensitivity, 87-percent specificity. DISCUSSION/CONCLUSIONS: Variability in identifying patients with stroke using ICD-9 codes has been reported in the literature and confirmed. Two coding algorithms for maximizing sensitivity or specificity are proposed. Caution is urged when using ICD-9-coded administrative data to identify patients with stroke.

Endodontic treatment in taurodontism with Klinefelter's syndrome: A case report.

Taurodontism occurs either as an isolated, singular trait or in association with syndromes and with some ectodermal anomalies. Successful endodontic treatment of taurodontism has rarely been reported. This article describes the successful treatment of a case of taurodontism with 5 canals. In addition, with the suggestion of taurodontism, as well as the clinical features of the patient, a tentative diagnosis of Klinefelter's syndrome was made. This diagnosis was proven after a chromosomal study. The discovery of taurodontism should alert the dentist that there may be associated systemic problems.

Dens invaginatus in the maxillary lateral incisor: treatment of 3 cases.

Nonsurgical treatment of 3 maxillary lateral incisors with dens invaginatus is reported. Invaginated teeth present technical difficulties with respect to their management because of complicated canal morphology. In the first case reported, apical abscess with incomplete root formation was found at the time of examination. Calcium hydroxide apexification provided favorable conditions for root closure and obturation. The other 2 cases were treated by conventional root canal treatment. Follow-up examinations showed successful clinical results, with osseous healing evident radiographically.

Outcomes of stroke patients in Medicare fee for service and managed care.

CONTEXT: Increasing numbers of Medicare beneficiaries have been enrolling in health maintenance organizations (HMOs) because HMO participation reduces out-of-pocket expenses, and the federal government views HMOs as a way to contain Medicare costs. However, results comparing outcomes and quality of care in HMOs vs fee for service (FFS) have been mixed, and outcomes after stroke have not been adequately assessed. OBJECTIVE: To compare discharge destinations and survival rates following stroke in Medicare HMOs with similar FFS settings. DESIGN: An observational study for 2 groups evaluating stroke patients' discharge destinations and survival times from the date of hospital admission. SETTING: A total of 19 HMOs were selected from 12 states. The FFS sample was drawn from the same geographic areas. PATIENTS: The sample included 402 HMO patients from 71 hospitals and 408 FFS patients from 60 hospitals. PROCESS AND OUTCOME MEASURES: Data were abstracted from medical records on demographics, clinical characteristics of stroke, comorbid illnesses, and discharge destinations following hospitalization. Data on survival were obtained from Medicare files and included 25 to 37 months of follow-up (median, 30.4 months, HMO; 31.1 months, FFS) from the date of hospital admission. RESULTS: There were 109 patients who died during the hospitalization (49 HMO, 12.2%; 60 FFS, 14.7%), and a total of 410 patients had died by the end of follow-up (191 HMO, 47.5%; 219 FFS, 53.7%). Approximately one fourth of both groups had do-not-resuscitate orders (HMO, 25.4%; FFS, 27.9%; P=.68). After controlling for age, marital status, and characteristics of dependency at discharge, HMO patients were more likely than FFS patients to be sent to nursing homes (HMO, 41.8%; FFS, 27.9%; P=.001) and less likely to be discharged to rehabilitation hospitals or units (HMO, 16.2%; FFS, 23.4%; P=.03). At follow-up, no significant differences in relative risk of dying were found between HMO and FFS groups (relative risk, 0.96; 95% confidence interval, 0.73-1.26; P=.77). CONCLUSIONS: Patients in Medicare HMOs who experience strokes are more likely to be discharged to nursing homes and less likely to go to rehabilitation facilities following the acute event. However, they have similar survival patterns compared with comparable patients in FFS settings after adjusting for other factors.

Lead acetate mutagenicity and mutational spectrum in the hypoxanthine guanine phosphoribosyltransferase gene of Chinese hamster ovary K1 cells.

The molecular nature of lead-induced mutations was examined in this study to more thoroughly understand lead mutagenesis. Chinese hamster ovary K1 cells were exposed to 0.5-3 mM lead acetate for 24 h. The median lethal dose (LD50) value was 1.5 mM, and the hypoxanthine guanine phosphoribosyltransferase (HPRT) mutant frequency increased linearly as lead concentrations were raised from 0.5 to 1.5 mM. We also amplified the HPRT cDNAs of 56 independent lead-induced mutants by reverse transcriptase-polymerase chain reaction (PCR). Forty-two mutant cDNAs were successfully amplified: 36 mutants had transcripts of normal or slightly smaller than normal size, and six mutants had large deletions. The other 14 mutants whose HPRT cDNA could not be amplified were subjected to genomic-DNA PCR analysis. All of those mutants had one or more exons missing from their genomic HPRT DNA. DNA sequencing of mutant cDNAs showed that 22 had single-base substitutions, four had small alterations, 10 had single-exon deletions, and six were missing two or three exons. Furthermore, DNA sequencing of the HPRT intron-exon boundaries in eight splice mutants revealed that all of them had single-base substitutions in their genomic DNA. G.C base substitutions occurred 3.3-fold more frequently than A.T base substitutions. Similar frequencies were observed for G.C-->A.T, G.C-->T.A, and G.C-->C.G mutations. These results suggest that G.C base pairs may be the primary target sites for lead mutagenesis.

Transcripts of metallothionein genes in Arabidopsis thaliana.

Using oligonucleotides with highly conserved cysteine-rich codons as probes, we isolated from a lambda YES Arabidopsis thaliana cDNA library two different sequences, each encodes an unique protein. These proteins, hereby designated as AtMT-q (AtMT-2) and AtMT-k (AtMT-1), are characteristic of metallothionein, carrying 13 and 14 cysteines in a total of 45 and 81 amino acids, respectively.

Nitrendipine in human plasma and breast milk.

To assess the disposition of the dihydropyridine calcium antagonist, nitrendipine, in lactating mothers, we studied three breast-feeding women to determine simultaneous plasma and breast milk concentrations of nitrendipine and its inactive pyridine analog metabolite after both a single 10 mg oral dose and 5 days of continuous therapy (20 mg per day). Nitrendipine was excreted in breast milk at peak concentrations ranging from 4.3 to 6.5 ng/ml 1-2 h after acute dosing while its inactive pyridine metabolite ranged from 6.9 to 11.9 ng.ml-1. After 5 days of dosing, Cmax remained in the same range and the breast milk/whole plasma concentration ratio for nitrendipine was 0.2 to 0.5. On the fourth day of continuous dosing, average concentrations of nitrendipine from 24-h collections of the milk were 1.1 to 3.8 ng.ml-1. Thus, nitrendipine and its metabolite are excreted in very low concentrations in human breast milk. Based on a maternal dose of 20 mg daily, a newborn infant would ingest an average of 1.7 micrograms of nitrendipine per day, or a relative dose of 0.095%.

The effects of age and race on nitrendipine pharmacokinetics and pharmacodynamics.

The pharmacokinetics and pharmacodynamics of a single 20-mg dose of nitrendipine (NTP) were studied in four groups of subjects (n = 9 per group). Group 1 were young white normotensive males, Group 2 were elderly white hypertensive males, Group 3 were black hypertensive males aged 40-55 years, and Group 4 were white hypertensive males aged 40-55 years. All other medications were withdrawn prior to NTP dosing. NTP was given in the morning 1 h after breakfast. Plasma samples for NTP assay were collected at predetermined times up to 48 h after dosing. Blood pressure was monitored before dosing, and at 0.5, 1, 3, 5, 12, and 24 h postdose. Pharmacokinetic parameters were found to be dependent on age. The area under the curve for Groups 1, 2, 3, and 4 was 50.5 +/- 19.4, 186 +/- 120, 107 +/- 49, and 88 +/- 43 ng h/ml, respectively (p less than 0.05). Corresponding values of elimination half-life were 9.9 +/- 1.3, 20 +/- 6.5, 13.3 +/- 6.1, and 15.9 +/- 8.0 h (p less than 0.05). Both diastolic and systolic blood pressures were significantly reduced from the baseline value in Groups 2, 3, and 4, with diastolic pressure remaining significantly lower than baseline at 24 h postdose. Based on the increased plasma levels and slower elimination of NTP in the elderly, as well as the measured blood pressure lowering, once daily dosing of NTP may be appropriate in some patients.

Gas and liquid chromatographic analyses of nimodipine calcium antagonist in blood plasma and cerebrospinal fluid.

Gas (GC) and liquid chromatographic (LC) assay procedures were developed for analysis of nimodipine (1,4-dihydropyridine calcium antagonist, BAY e 9736) in blood plasma at low nanogram concentration levels. To avoid decomposition during gas chromatography, nimodipine was oxidized to nimodipine pyridine (P) analogue before it was chromatographed on the OV-17 column and quantitated using an electron-capture detector. In contrast, the LC procedure involved chromatographic separation and quantitation of the underivatized nimodipine and of the endogenous P analogue using a 3-micron Spherisorb ODS column and UV detection. The same plasma extract and three alternative internal standards were used for both assays. Taking into consideration the fact that the GC assay result includes endogenous P analogue as well as nimodipine, good correlation between GC and LC assay data was obtained. Comparison of the results observed with the two procedures confirmed the accuracy of each procedure and provided an alternative when one of the assay results was subject to patient plasma constituent interference. The LC assay was also used for analysis of the demethylated metabolites of nimodipine. To detect sub-nanogram concentrations of nimodipine in cerebrospinal fluid a combined LC-GC procedure using an LC clean-up step and a GC quantitation step was also developed. The above GC and LC procedures were used to obtain preliminary pharmacokinetic data.