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Background: Increased levels of plasma nuclear and mitochondrial DNA have been reported in critically ill patients. We tested the hypothesis that plasma nuclear and mitochondrial DNA are substantially increased in acute bacterial meningitis and decrease after antimicrobial therapy, and that plasma nuclear and mitochondrial DNA levels can predict treatment outcomes., Methods: We examined serial plasma nuclear and mitochondrial DNA levels in 22 adult community-acquired bacterial meningitis (ACABM) patients. The plasma nuclear and mitochondrial DNA levels were also evaluated in 11 aseptic meningitis patients and 22 volunteer subjects during the study period., Results: All of the both bacterial and aseptic meningitis groups had a higher plasma DNA levels on admission as compared with those of volunteer groups. Levels of plasma nuclear and mitochondrial DNA in ACABM cases were significantly increased initially and substantially decreased thereafter. Both plasma nuclear DNA and plasma mitochondrial DNA levels at presentation are significantly negative correlate with modified Barthel Index (average) (r = -0.639, P = 0.004 and r = -0.551, P = 0.018) at 3 months after discharge (average), respectively, in this study. Both higher plasma nuclear (cutoff value of >169 ng/ml) and mitochondrial DNA levels (cutoff value of >58.9 ng/ml) at presentation were associated with poor outcome in ACABM patients., Conclusion: Based on our results, the higher plasma DNA levels were associated with a poorer outcome. Therefore, we look forward to more prospective multicenter investigations specifically to confirm the predictive value of plasma DNA levels in outcome prediction.

The clinical data and cerebrospinal fluid (CSF) 14-3-3-gamma protein detection of eight adult HIV-negative cryptococcal meningitis (CM) cases were examined. The eight cases included six males and two females aged 35-70 years (mean = 49.8 years). The duration between the onset of CM symptoms and the first CSF study ranged from 1 to 60 days. Initial neuroimaging study was abnormal in 87.5% (7/8) of the cases. All the eight had positive initial and subsequent follow-up CSF 14-3-3-gamma protein detection. The densitometric values of CSF 14-3-3-gamma protein were not correlated with either the CSF white blood cell counts or the therapeutic results. The therapeutic results showed that three cases died and five survived. Significant neurologic deficits were shown in 60% (3/5) of the survivors. This study revealed that HIV-negative CM patients have elevated CSF 14-3-3-gamma protein levels, and that this level is not changed with a short-term treatment.

The effects of Herpes simplex virus 1 (HSV-1) infection on five different types of oral cancerous cells (neck metastasis of gingival carcinoma (GNM) cells and tongue squamous cells of carcinoma (TSCCa) and non-cancerous cells (buccal mucosal fibroblasts (BF), gingival fibroblasts (GF), oral submucosal fibrosis cells (OSF)) and one type of non-oral cancerous cells (KB cells) were investigated. In HSV-1-infected cells the cell viability, CPE, viral antigens accumulation, caspase-3 activity, annexin V binding and DNA fragmentation were estimated. Three different forms or pathways of cell death were considered: apoptosis (the presence or rise of caspase-3 activity, DNA fragmentation and annexin V binding), slow cell death (the presence or rise of DNA fragmentation, the absence or decline of caspase-3 activity and annexin V binding), and necrosis (the absence of decline of caspase-3 activity, DNA fragmentation and annexin V binding). The viability of all cell types, except for KB cells, was reduced by the infection. CPE and viral antigens data demonstrated that all six types of cells could be infected with HSV-1. Upon HSV-1 infection there occurred (i) a classical apoptosis in GF cells, (ii) apoptosis in the early phase of infection and necrosis in the late phase of infection in GNM and TSCCa cells, (iii) slow cell death followed by necrosis in BF and OSF cells (however, these cells showed a different type of CPE), (iv) a classical slow cell death in KB cells. It is hypothesized that HSV-1 infection has a potential to induce several distinct pathways leading to cell death or several forms of cell death. Moreover, more than one pathway may be involved in the death of particular cell type. As HSV-1 was demonstrated to infect different oral and non-oral cells and cause different pathways or forms of cell death, the safety of using HSV-1 as a vector for gene therapy should be re-considered.

Problems associated with polynuclear aromatic hydrocarbon (PAH) contaminated site in environmental media have received increasing attention. To resolve such problems, innovative in situ methods are urgently required. This work investigated the feasibility of using surfactants to extract phenanthrene on spiked sand in a batch system. Phenanthrene was spiked into Ottawa sand to simulate contaminated soil. Six surfactants, Brij 30 (BR), Triton X-100 (TR), Tergitol NP-10 (TE), Igepal CA-720 (IG), sodium dodecyl sulfate (SDS) and hexadecyl trimethyl ammonium bromide (HTAB) were used. Adjusting the extraction time, mixing speed and surfactant concentration yielded the optimum extracting conditions. The concentration of phenanthrene was identified with HPLC. Under the experimental conditions, results indicated that those surfactants were highly promising on site remediation since the residual phenanthrene concentration was effectively reduced. The optimum operating conditions were obtained at 30 min, 125 rpm and surfactant concentrations in 4%.

We report a 38-year-old male patient with rare concurrent intracranial and intramedullary tuberculoma demonstrated by serial magnetic resonance image (MRI) and confirmed by a pathological study. The involvement of the middle cerebral artery is also shown by magnetic resonance angiography. These findings have a good correlation with the clinical features including cranial nerve involvement, stroke-like symptoms, and paraplegia.

The cranial computed tomography (CT) and magnetic resonance image (MRI) studies of three Japanese encephalitis (JE) patients, 24 to 37 years of age, are reported. The initial findings of CT study were limited but initial MRI studies revealed multiple lesions involving the brainstem, basal ganglia and bilateral thalami. Follow-up MRI studies showed small residual lesions only. The result shows that MRI can delineate and detect brain lesions better than CT in patients in the acute stage of JE. The locations of lesions in MRI study are noteworthy and have a good correlation with pathologic anatomic distribution. Therefore, MRI study is helpful in early diagnosis of JE.

In response to the need for rapid screening of combinatorial libraries to identify new lead compounds during drug discovery, we have developed an on-line combination of ultrafiltration and electrospray mass spectrometry, called pulsed ultrafiltration mass spectrometry, which facilitates the identification of solution-phase ligands in library mixtures that bind to solution-phase receptors. After ligands contained in a library mixture were bound to a macromolecular receptor, e.g., human serum albumin or calf intestine adenosine deaminase, the ligand-receptor complexes were purified by ultrafiltration and then dissociated using methanol to elute the ligands into the electrospray mass spectrometer for detection. Ligands with dissociation constants in the micromolar to nanomolar range were successfully bound, released, and detected using this method, including warfarin, salicylate, furosemide, and thyroxine binding to human serum albumin, and erythro-9-(2-hydroxy-3-nonyl)adenine binding to calf intestine adenosine deaminase. Repetitive bind- and-release experiments demonstrated that the receptor could be reused. Thus, pulsed ultrafiltration mass spectrometry was shown to provide a simple and powerful new method for the screening of combinatorial libraries in support of new drug discovery.

Background: Resection for hepatocellular carcinoma in patients with cirrhosis and impaired liver function is usually unjustified because of higher surgical risks and poorer long-term prognosis., Methods: A retrospective comparison of the background and resectional results of patients with cirrhosis and hepatocellular carcinoma was carried out between those with preoperative indocyanine green 15-minute retention rate of 20% or greater (group A, impaired function group, n = 36) and those with indocyanine green 15-minute retention rate of 10% or less (group B, normal function group, n = 34)., Results: The group A patients had significantly lower serum albumin level and higher serum bilirubin level, longer prothrombin time, higher incidence of associated esophageal varices, and poorer Child's classifications for cirrhosis. Although the tumor diameter in both groups was similar (A, 6.9 versus B, 7.1 cm; p = 0.94), the resected liver amount in group B was greater (227.4 versus 473.2 gm; p = 0.038) because of a greater extent of liver resection (p < 0.001) and a wider surgical margin (0.34 versus 1.85 cm; p < 0.0001). The amount of operative blood loss and blood transfusion, operative morbidity, and operative mortality were not different between the two groups. The pathologic characteristics and staging were also comparable. The 5-year disease-free and actuarial survival rates of groups A and B were 30.9% and 29.6% (p = 0.16) and 45.2% and 33.4% (p = 0.11), respectively., Conclusions: If the amount of resected nontumorous liver parenchyma could be reduced, resection of hepatocellular carcinoma in selected patients with cirrhosis and impaired liver function is still justified in spite of a narrow surgical margin.

High-performance liquid chromatography (HPLC)-electrospray mass spectrometry (LC-MS) was used to analyze vitamin A-active retinoids including retinoic acid, retinol, retinal, and retinyl acetate. Unlike previous LC-MS methods such as negative ion electron capture chemical ionization, no derivatization of retinoic acid was required. HPLC separations were carried out on a C30 reversed phase column with gradient elution using mobile phases containing water, methanol, and methyl-tert-butyl ether. Ammonium acetate (5 mM) was added to the mobile phase to facilitate ion pair formation during reversed phase HPLC of retinoic acid, and acetic acid (0.5% v/v) was added to the mobile phase to enhance protonation during LC-MS analysis of nonacidic retinoids. During negative ion electrospray, retinoic acid formed abundant deprotonated molecules, [M-H]-, of m/z 299 without significant fragmentation. Although retinol, retinal, and retinyl acetate did not ionize during negative ion electrospray, the positive ion electrospray mass spectra of these retinoids showed an abundant protonated molecule of m/z 285 for retinal and base peaks of m/z 269 corresponding to elimination of water or acetic acid from the protonated molecules of retinol or retinyl acetate, respectively. No ions from retinoic acid were detected during positive ion electrospray. Limits of detection for retinoic acid, retinal, retinol, and retinyl acetate were 23 pg, 1.0 ng, 0.5 ng, and 10 ng, respectively.

Fluorescence decay profiles of four oligonucleotide duplexes, [table: see text] ((+)- and (-)-trans-1) and [table: see text] ((+)- and (-)-trans-2), in which an exocyclic amino group of deoxyadenosine (A*) or deoxyguanosine (G*) has been alkylated by trans opening at C-10 of the epoxide group of either the (+)-(R,S,S,R)- or (-)-(S,R,R,S)-enantiomer of (+/-)-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10- tetrahydrobenzo[a]pyrene (BPDE in which the benzylic 7-hydroxy group and the epoxide oxygen are trans), exhibit more than one fluorescence lifetime. Decay profiles of the oligomers, measured at 15 degrees C with excitation and emission wavelengths of 335 and 400 nm, respectively, have been analyzed using a triple-exponential decay law. Results for (+)- and (-)-trans-1 and -2 have been compared with results for the modified, single-stranded oligonucleotides ((+)- and (-)-trans-SS-1, and (+)- and (-)-trans-SS-2) and for the cis and trans opened products formed on alkylation at the 6-amino group of 2'-deoxyadenosine 5'-phosphate by (+)-(R,S,S,R)-BPDE ((+)-trans- and (+)-cis-A). The profiles of (+)-trans- and (+)-cis-A are well represented by single-exponential decay laws with lifetimes of 86 and 110 +/- 3 ns, respectively. For the single- and double-stranded oligomer adducts, which exhibit at least three fluorescence lifetimes, two of the lifetimes are short (0.5-14 +/- 1 ns) and one is long (35-59 +/- 3 ns). The fluorescence lifetimes and the amplitudes of the long-lived components in the decay profiles of the double-stranded oligomer adducts are generally smaller than those for the corresponding single-stranded adducts. The data provide evidence that the double-stranded oligomer adducts exist as multiple conformations. Previously reported NMR results suggest that the short lifetime fluorescence components are due to major adduct conformations in which the pyrenyl group is intercalated ((+)- and (-)-trans-1) or lies in the minor groove ((+)- and (-)-trans-2). The observation of long lifetime fluorescence species for the double-stranded oligomers is consistent with the presence of minor conformations (approximately 1-5%) in which the double-stranded oligomer either is locally denatured or is a mixture of locally denatured double-stranded conformations and equilibrium concentrations of single-stranded oligomers.

Closed-circular, single-stranded viral DNAs are widely employed in DNA cloning and sequencing experiments. Because of their well-defined structure and sequence, closed-circular, single-stranded DNAs have also been used for ligand binding experiments and light scattering measurements. However, there is a high molecular weight impurity observed in light scattering experiments, which sometimes contaminates single-stranded DNA purified from phage that has been precipitated in polyethylene glycol, average molecular weight 8000, and purified by standard phenol-chloroform extraction. Three methods have been examined that remove this impurity from closed-circular, single-stranded M13mp19 DNA (SS M13 DNA). One employs a commercial peparation. This procedure yields pure but degraded SS M13 DNA, as shown by light scattering measurements and HPLC. Another employs a Whatman DE52 (diethylamino cellulose) column. This procedure yields intact DNA, but in poor yield (less than 20% of that obtained by phenol-chloroform extraction). The last was the most successful. This employs HPLC with a Waters AP-1 column with DEAE 8HR bedding. This procedure, which provides DNA in high yield (80%-90% column recovery) with an intact structure, is an efficient method for the isolation of high-purity, closed-circular, single-stranded viral DNA suitable for physical investigations and ligand binding measurements.

From 1967 to 1992, 39 patients with urogenital rhabdomyosarcoma were admitted to our hospital. Of them 30 were male and 8 female and their age ranged from 4 months to 14 years (32 were less than 5 years). Primary tumor was located in bladder or/and prostate in 26, paratesticule 9, vagina 3, and penis 1. Among the 31 patients receiving treatment, 23 were followed up. There were 15 (65%) survivors, including 13 who were free from tumor for over 4 years. The eldest two patients were 20 and 21 years of age. Two patients with vaginal tumor and 4 with bladder tumor preserved their organs with normal function. Biopsy were done only in 8 patients. In recent years, conservative surgery combined with chemotherapy for urogenital rhabdomyosarcoma has been justified.

A DNA fragment containing Epstein-Barr virus (EBV) terminal fragment sequence was obtained from a genomic library of nasopharyngeal carcinoma (NPC). One of the clones (clone 1510) contained the gene encoding latent membrane protein (LMP). Sequence analysis revealed that this gene had 95% homology with the LMP sequence of the B95-8 strain. Among the sequence variations, there was a change from G to T at nucleotide position 169,426, resulting in the loss of an XhoI site in exon 1 of the LMP gene. A pair of primers bracketing the XhoI site were designed to synthesize the EBV DNA fragment from nucleotides 169,081-169,577 by using the polymerase chain reaction (PCR) method. The PCR products were then subject to XhoI digestion and to DNA sequencing analysis. This restriction enzyme site polymorphism along with the sequence variations were also observed in 50 biopsy tissues as well as in the throat washings of 6 out of 20 healthy individuals that we examined, indicating that the EBV strain predominantly existing in these biopsy tissues was different from strains of B95-8, Jijoye or nude mouse passaged cells (C15) with an African origin, but closely resembled other nude mouse passaged CAO cells which were originally derived from China. Balb/c 3T3 cells carrying this NPC-LMP gene showed a transformed cell morphology and were tumorigenic in nude mice. The relationship between this unique type of EBV and NPC has yet to be established.

In a stratified multi-stage random sample, 1600 persons of 60 years and older in Shanghai, People's Republic of China, were investigated for periodontal conditions using the CPITN method. Periodontal conditions seemed to deteriorate steadily but rather slowly with increasing age. In the older age groups, there was a trend for an increased proportion of both subjects and sextants scoring CPITN code 4 (deep pockets). The increase in tooth loss with increasing age was more clearly visible: increasing percentages of edentulousness and decreasing mean numbers of teeth remaining in dentate subjects.

The rheological properties of whole human blood exhibit thixotropic behavior at low shear rates up to about ten reciprocal seconds (1). The accepted cause of this shear rate-dependent and time-dependent behavior is the progressive breakdown of rouleaux into individual red cells. Huang developed a rheological equation which incorporates the kinetics of rouleau breakdown in his models (2). This five-parameter equation was used successfully to represent the hysteresis loop and the torque-decay curve of whole human blood. Numerical values of these five thixotropic parameters, which characterize the rheological behavior of the blood from apparently healthy human subjects, were established (3). In this communication, we examined the effect of hematocrit on each of the above mentioned parameters. The results show that the following parameters will increase their values with an increase in hematocrit: the yield stress, Newtonian contribution of viscosity, non-Newtonian contribution of viscosity, apparent viscosity and the equilibrium value of the structural parameter which indicates the relative amount of rouleaux in blood. Mathematical equations were developed to give the relationship between parameters and hematocrit. Two other thixotropic parameters, viz. the kinetic rate constant of rouleaux breakdown into individual red cells and the order of the breakdown reaction, were found to be independent of the hematocrit. It is consistent with reaction kinetic theory that the rate constant and the order of reaction are independent of the concentration of reactants.

The steady state non-Newtonian viscosity of whole human blood has been widely studied as a function of the shear rate; and used to characterize the blood in various pathological disorders. In our previous studies, we demonstrated that blood is a thixotropic fluid. Its time-dependency and shear rate dependency of rheological behavior can be represented by an equation developed by Huang. Parameters of the equation can be used for the characterization of an individual's blood. They provide information, such as the kinetic rate constant of breakdown of RBC rouleaux to individual erythrocytes and the relative amount of rouleau formation in the dynamic equilibrium between rouleaux and individual erythrocytes. In this communication, the thixotropic parameters from blood samples of fifteen apparently healthy human subjects were investigated. When compared to the use of apparent viscosity values for the correlation with a pathological disorder, thixotropic parameters are preferable. The mean values of thixotropic parameters obtained from apparently healthy human subjects provide a base for comparison with the same parameters as obtained from blood samples of patients with certain pathological disorders involving the circulation.