Low-temperature femtosecond-resolution transient absorption spectroscopy of large-scale symmetry mutants of bacterial reaction centers

Reaction centers isolated from three large-scale symmetry mutants sym0, sym2-1, and sym5-2 described in the previous article of this issue [Taguchi, A. K. W., Eastman, J. E., Gallo, D. M., Jr., Sheagley, E.. Xiao, W., & Woodbury, N. W. (1996) Biochemistry 35, 3175-3186] have been investigated by low-temperature ground state and ferntosecond-resolution transient absorption spectroscopy. All three of these large-scale symmetry mutants undergo electron transfer at 20 K. The mutants sym0 and sym5-2 have yields and dominant rates of charge separation comparable to wild type. However. the sym2-mutant shows a roughly 35%, quantum yield at this temperature, and the major kinetic component of the initial electron transfer is slower than wild type by nearly a factor of 100. The sym0 mutant showed substantial changes in the monomer bacteriochiorophyll ground state and transient spectra, and both sym0 sym2-1 showed changes in the bacteriopheophyll ground state and transient spectra. In particular, sym2-1 shows a small absorbance decrease in the region of the Qx band of the B side bacteriopheophytin which could be attributed to 10%-20% electron transfer along the B pathway.     

Canine excretory urogram: correlation with base-line measurements

Ten healthy, adult mongrel dogs were each given various dose levels of sodium iothalamate IV, and postinjection radiographs were made. Base-line plasma and urine osmolality, glomerular filtration rate, packed cell volume, plasma protein concentration, serum urea nitrogen concentration, serum creatinine concentration, and urine specific gravity were measured. The base-line values and the dose of contrast medium were compared statistically with linear and density measurements made from the post-injection radiographs. Base-line plasma osmolality was directly related to the pyelographic and nephrographic density. The dose of contrast medium directly influenced the kidney length, the kidney, pelvic, diverticular, and ureteral widths, and the renal and diverticular densities despite variations in base-line values within accepted limits.     

Tissue optical property measurements: overestimation of absorption coefficient with spectrophotometric techniques

We have cloned both human and murine complementary DNAs that are homologous to the Drosophila serine/threonine polo kinase and the recently cloned murine polo related kinase (PLK). Both the human and murine clones are about 2.1 kilobases with open reading frames of 1.8 kilobases, encoding proteins of 603 amino acids with a predicted size of 66 kilodaltons and an apparent size of 67 kilodaltons by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. During embryonic development of the mouse, the mRNA was expressed in all tissues examined, whereas in adult tissues, expression was limited to thymus and ovaries. All cell lines examined also expressed mRNAs of similar size. Microinjection of in vitro transcribed sense mRNA into serum-starved murine NIH3T3 cells induced tritiated thymidine incorporation, whereas microinjection of antisense RNA into growing NIH3T3 cells blocked tritiated thymidine incorporation. When PC12 rat cells were induced to differentiate with nerve growth factor, gene expression of PLK was greatly reduced. Together, these results suggest that PLK expression is restricted to, and is perhaps required by, proliferating cells.     

Mechanism of the alpha-complementation reaction of E. coli beta-galactosidase deduced from fluorescence correlation spectroscopy measurements

Glycoprotein K (gK) is involved in membrane fusion phenomena during infectious virus production and egress and is an important determinant for neurovirulence. To assess better the in vitro and in vivo roles of gK in virus replication, a recombinant virus was constructed expressing an engineered enhanced green fluorescent protein (EGFP) under the control of the human cytomegalovirus immediate early gene promoter (HCMV-IEP) inserted in place of the gK gene. The EGFP gene insertion was confirmed by diagnostic polymerase chain reaction (PCR), and the presence of the EGFP protein was detected by western immunoblot analysis using anti-GFP monoclonal antibody. Fluorescence microscopy revealed that virus infected cells emitted bright fluorescence when examined using filters for fluorescein. Fluorescence emission was detected as early as 4 h post-infection. Fluorescence intensity increased over time and was stable at late times after infection at which point viral plaques continued to emit bright green fluorescence. The amount of fluorescence emitted by virus infected Vero cells was monitored by fluorescence cytometry using a FACS cytometer. At an MOI of 3, all infected cells emitted strong green fluorescence as quantified by cytometry at 48 h post-infection. The deltagK-EGFP expressing recombinant virus will enable the determination of the role of gK in virus entry and egress as well as the role of gK in the molecular pathogenesis of herpes simplex virus type 1 (HSV-1).     

Immunohistochemical metallothionein expression in thymoma: correlation with histological types and cellular origin

In radioimmunotherapy, the long circulation times of antibody radioconjugates correlate with high relative radiation doses to nontumor tissues. Tumor/normal tissue ratios can be significantly improved by using targeting molecules with shorter circulation times. IFabs are multimers of VH-CH1-linker-VK-CK monomers. The lack of the Fc region in IFabs should lead to circulation times that are shorter than those of IgG molecules. The monomers assemble into disulfide-bond-stabilized multimers, 90% of which are 100 kDa dimers (IFab2). IFab2s should not be rapidly eliminated through kidney filtration because their molecular weight is above the threshold for renal passage. We report the first experimental in vivo tests for 125I-IFab radioconjugates derived from a humanized version of the anti-breast mucin monoclonal antibody BrE-3. Biodistributions are reported for athymic nude mice carrying human mammary tumor MX-1 xenografts. The T1/2 beta's for the different tissues ranged from 13.3 h for blood to 19.9 h for tumor. Therefore, IFab radioconjugates cleared the body with a rate comparable to that of F(ab')2 fragments. Except for stomach, tumor/nontumor dose ratios were significantly better for IFabs than for the parent antibody (BrE-3)4 days after injection.     

Intracranial tuberculomas: MRI signal intensity correlation with histopathology and localised proton spectroscopy

In seven cases of intracranial tuberculomas showing different signal intensities on MRI (five characteristic and two nonspecific), detailed histopathological examination was performed to look for number of macrophages, fibrosis, gliosis, degree of inflammatory cellular infiltrate, and type of caseation. The granulomas showing more macrophages, fibrosis and gliosis appeared hypointense on T2-weighted images. Tuberculomas showing minimal macrophages, marked cellular infiltration, and minimal fibrosis appeared hyperintense on T2-weighted images. Lesions showing similar signal intensity on T2-weighted images showed variation in the amount of macrophages, cellular infiltrates, maturity and fibrosis. Trace element estimation was done (iron, copper, and magnesium) in two of these lesions appearing hypointense on T2 and two normal brain samples; these were significantly lower in tuberculoma compared to normal brain tissue. Localised proton spectroscopy was performed in two hypointense lesions which showed marked increase in peaks in the region of mobile lipids (1.28 ppm) compared to normal brain parenchyma. It is concluded that the signal intensity of the lesions is dependent on the number of macrophages, fibrosis and cellular infiltrates. In addition increased lipid contents in the tuberculoma also contribute to the hypointensity on T2-weighted images.     

火焰原子吸收光谱法测定矿石中锑的不确定度评定

针对火焰原子吸收光谱法测定矿石样品中锑的分析测定过程和数学模型,找出了影响测量不确定度的主要来源,对各不确定度分量进行了评定,并计算得到了合成标准不确定度和扩展不确定度,给出了相应的测定结果表示法。     

Radiographic assessment of hyperinflation: correlation with objective chest radiographic measurements and mechanical ventilator parameters

BACKGROUND: Pulmonary barotrauma is a potentially fatal complication of positive pressure ventilation. We previously found that barotrauma occurred in patients with radiographic hyperinflation, but few objective data define the relationships among hyperinflation, objective chest radiograph (CXR) measurements, ventilator parameters, and development of barotrauma. OBJECTIVES: We sought (1) to assess the relationships among hyperinflation, objective CXR findings, mechanical ventilator parameters, and development of barotrauma. (2) To compare radiographic hyperinflation, ventilator parameters, and incidence of barotrauma in a current group of ICU patients with historical control subjects. SETTING: Medical and surgical ICU patients in a university hospital. DESIGN: Prospective blinded observational study; comparison of current series with historical control subjects. METHODS: One hundred two prospectively enrolled mechanically ventilated medical and surgical ICU patients each received portable supine CXRs that were reviewed independently by three radiologists who made objective measurements and subjectively determined the likelihood of hyperinflation. Ventilator parameters were recorded at the bedside at the time each CXR was obtained. CXR measurements and ventilator parameters were then related to the development of barotrauma during the course of ventilation and compared with findings of a prospective study at our institution 1 year earlier. RESULTS: Radiographically recognizable hyperinflation occurred in 18 of 102 mechanically ventilated ICU patients (18%) and correlated with lung length (24.7 vs 19.8 cm; p<0.05) and the anterior rib number that intersects the hemidiaphragm (5.4 vs 4.7; p<0.05). Patients with hyperinflation were ventilated at higher tidal volume per kilogram (VT/kg) (11.0 vs 9.4; p=0.0081), but peak airway pressure, plateau pressure, and positive end-expiratory pressure were similar. There were significant decreases in VT (810 vs 739 mL; p=0.015) and VT/kg (11.0 vs 10.1 mL/kg; p<0.001) in these mechanically ventilated ICU patients in comparison to hospital control subjects evaluated during the previous year. Paralleling these changes was a decrease in the frequency of CXR hyperinflation (p=0.003) and the incidence of ventilator-associated barotrauma (6.5% vs 0.98%; p=0.048). CONCLUSIONS: Ventilation at higher VT/kg is associated with a higher incidence of CXR hyperinflation. Radiographic hyperinflation is associated with lung length > or =24.7 cm and visualization of the sixth anterior rib. Patients with hyperinflation may be at greater risk for developing barotrauma or volutrauma. Ventilatory strategies utilizing lower volumes are associated with a lower incidence of such trauma in the current sample as compared with historical control subjects.     

Vaccination of racing greyhounds: effects on humoral and cellular immunity

Greyhound kennel owners frequently employ multiple vaccination schedules in an attempt to reduce financial losses incurred as a result of infectious diseases. In order to determine the effects of multiple vaccination schedules on the immune system of racing greyhounds, three litters of greyhound pups raised in laboratory conditions were divided into two groups and subjected to either a maximum or a minimum vaccination schedule. Blood samples were collected biweekly for 6 months beginning at 2 weeks of age and analyzed to establish 'baseline' values for the lymphatic system of greyhounds. Lymphocyte transformation, total and differential leukocyte counts, and flow cytometry were used to evaluate cellular immunity. Humoral immunity was evaluated using serum neutralization and hemagglutination inhibition tests. Proliferation of peripheral blood lymphocytes in response to the mitogen concanavalin A (Con A) was higher for the maximum vaccination groups. The frequency distribution of circulating CD4 and IgG labeled lymphocytes was higher in the minimum vaccination groups. A significant treatment by time interaction in CD4, IgG, and IgM labeled cells was observed, This interaction, however, was not significant at any point in time for CD4 and IgG labeled cells. The percentage of lymphocytes expressing surface IgM was significantly higher in the minimum vaccination groups at 10 and 14 weeks of age. No significant differences were detected in humoral immunity between the maximum and minimum groups of each litter. Results of this study indicate that maximum vaccination schedules do not appear to be more effective or more immunosuppressive than minimum vaccination schedules.     

原子吸收法快速测定金精矿中的银

采用王水+H2O2+NH4Cl+HF溶样体系封闭溶样法对金精矿进行预处理,制得样品溶液后以原子吸收法测定银;其测定范围1000 g/t,RSD为4.5%,测定60个样品仅需3 h。该方法具有简单快速、成本低、对环境友好等特点,其分析结果可满足选冶工艺生产的要求。     

The effects of food on methotrexate absorption

OBJECTIVE: To determine the effects of food on methotrexate (MTX) absorption in patients receiving MTX for the treatment of rheumatoid arthritis (RA). METHODS: Standard pharmacokinetic variables were determined in patients with RA after their usual maintenance dose of MTX, under fasting conditions and after they ate a standard breakfast. RESULTS: No significant differences in area under the serum concentration versus time curve, maximal MTX concentration after dosing (Cmax), time to Cmax), bioavailability, urinary MTX, renal clearance of MTX, or creatinine clearance were observed between the 2 dosing conditions. CONCLUSION: We observed no significant effect of food on MTX absorption or bioavailability. Patients may consume MTX without regard to meals.