Poly[hexa-aqua-copper(II) [di-µ(3)-sulfato-disodiate(I)]].

The title compound, {[Cu(H(2)O)(6)][Na(2)(SO(4))(2)]}(n), has been prepared under mild hydro-thermal conditions and has been structurally characterized. It exhibits a structure in which the inorganic frameworks are three-dimensional, participating in extensive hydrogen bonding. Copper occupies a special position (). The Na atom is coordinated by five O atoms of four sulfates [Na-O distances are between 2.825 (3) and 2.983 (3) Å]. The four O atoms of the sulfate ligand are coordinated to four Na atoms, the sulfate ligands coordinating in a chelating/bridging tetra-dentate mode.

An interpolated Markov model polishes Gibbs sampling's ability in detecting regulatory elements.

Microarray techniques provide new methods to find coregulated genes based on their coexpression profiles. Under the assumption that coregulated genes share cis acting regulatory elements, it is important to investigate the upstream sequences controlling the transcription of these genes. A modified Gibbs sampling algorithm with background interpolated Markov model (IMM) has been developed to detect regulatory elements in the upstream regions of translation start site of coexpressed genes. Simulated data are used to test our algorithm successfully. Results show that the improved Gibbs sampling has better performance in extracting less-conserved elements than algorithms with single nucleotide independent model and fixed higher-order Markov models. Then, upstream sequences of two clusters of coexpressed genes from Saccharomyces cerevisiae under diauxic shift conditions are analyzed, several putative motifs that may be involved in the pathway are found.

[Studies on technology of supercritical-CO2 fluid extraction for volatile oils and saikosaponins in Bupleurum chinense DC].

OBJECTIVE: To study the technology of supercritical-CO2 fluid extraction (SFE-CO2) for the volatile oils and saikosaponins in Bupleurum chinense. METHOD: Exploring the effects of pressure, temperature, extraction time, flow rate of CO2 and entrainers on the yield of the oils and saikosaponin-contained extracts; determining the optimum conditions for SFE-CO2; analyzing the oils by GC/MS and comparing the technology of SFE-CO2 with that of traditional steam distillation. RESULT: The optimum extraction conditions turned out to be--for volatile oils: pressure (EP) = 20 MPa, temperature (ET) = 30 degrees C, isolator I pressure (1P-I) = 12 MPa, temperature(1T-I) = 65 degrees C, isolator II pressure (1P-II) = 6 MPa, temperature (1T-II) = 40 degrees C, extraction time = 4 hours, and CO2 flow rate = 10-20 kg.(h.kg)-1 crude drug; for saikosaponins: EP = 30 MPa, ET = 65 degrees C, 1P I = 12 MPa, 1T I = 55 degrees C, 1P II = 6 MPa, 1T II = 43 degrees C, extraction time = 3 hours, entrainer = 60% ethanol, and CO2 flow rate = 20-25 kg.(h.kg)-1 crude drug. CONCLUSION: SFE-CO2 excels the traditional steam distillation in raising yield and reducing extraction time. The oils are composed of 22 constituents including caproaldehyde, and the saikosaponins can only be extracted with the help of entrainers under higher pressure and temperature.

Immunogenic potential of rgp120 from African HIV-1 subtype A.

Previous studies have shown that the African strains of HIV-1 mostly cluster with the subtypes A, C or D based on phylogenetic analysis of the ENV nucleotide sequences. In the present investigation we have examined the immunogenic potential of full length gp120 derived from the Ugandan HIV-1 subtype A isolate, AUG06c, using computer-based prediction methods and a plasmid-mediated immunization technique. Computer-assisted analysis of the amino acid residues identified 15 potential B-cell epitopes in gp120 of AUG06c. Despite marked variation in the primary sequences, these epitopes were shown to correspond well to analogous sites in gp120 derived from the subtype B reference clones, MN and IIIBBH10. The relative positions of the epitopes indicated that E9[V3], E14[C3] and E15[V5] correspond to the previously defined principal neutralizing determinant (PND) located in the V3 loop, the CD4 binding site and gp120 "immunodominant" region, respectively. Intramuscular inoculation of BALB/c mice with the ENV clones from AUG06c or from the subtype C clone, CUG045 elicited antibodies which react with the homologous but not with the heterologous PND peptide in ELISA. However, cocktail inoculation with the ENV plasmids from AUG06c and CUG045 elicited antibodies which reacted with both peptides. Antibody response to the other predicted epitopes of AUG06c was not as strong as the response to the PND peptide. The response of the mice to DNA-mediated immunization was further tested in a proliferation assay. Spleen cells derived from the immunized mice exhibited a strong proliferative response to homologous and heterologous PND peptides in [3H]thymidine incorporation assay. DNA-mediated immunization with rgp120 of AUG06c appears to elicit cellular immune response of relatively broad specificity.

Seroreactivity of analogous antigenic epitopes in glycoprotein 120 expressed in HIV-1 subtypes A, B, C, and D.

This article describes the impact of sequence variation on the distribution and seroreactivity of linear antigenic epitopes in gp120 encoded in new Ugandan HIV-1 clones from subtypes A, C, and D, and in North American clones from the B subtype. A region of the env gene encoding the C2 to V5 domains was PCR amplified from the lysates of peripheral blood leukocytes or from short-term cultured isolates. Computer-assisted analyses were conducted on the amino acid sequences to determine the distribution of surface structures in gp120. Despite marked sequence diversity, eight analogous epitopes were predicted for all clades of the virus analyzed. Synthetic peptides comprising the putative principal neutralizing determinant E2[V3], and other B cell epitopes E3[V3-V4], E4[V3-V4], E7[C3], and E8[V5], from a seroprevalent Ugandan isolate, AUG06c, were tested in ELISA for antigenicity with sera from Uganda, New York, and Thailand. Variable magnitudes of seroreactivity were observed for all of the peptides tested. However, a significantly higher degree of serum cross-reactivity was detected with the V3 loop peptide. ELISA reactivities of the same serum panel indicated that V3 loop peptides containing the apical residues GPGR (clones AUG06c and BRT3) or GPGQ (CUG045 and DUG044) were more antigenic and display extensive cross-reactivity as compared to analogous peptides comprising GLGQ (DUG23c), GQGQ (DUG042), or GPWG (BRT1). BETATURN analysis of the divergent V3 loop apical residues showed a good correlation of probable beta-turn occurrence with strong seroreactivity. These findings suggest that the major antigenic specificities in the divergent clades of HIV-1 are well conserved.(ABSTRACT TRUNCATED AT 250 WORDS)

Relative reactivity of the V3 loop PND of HIV-1 subtypes A, B, C, D, and F with sera from selected Ugandan localities.

Synthetic peptides comprising the predicted principal neutralizing determinant (PND) in new African and North American HIV-1 clones were tested in ELISA for reactivity with ninety six serum samples from asymptomatic donors in six selected localities in Uganda. Irrespective of the geographical origin of the samples, the majority of the test sera cross-reacted at high intensities with the peptides derived from the North American clone, BRT3.6 (Group B), the Ugandan clone, CUG045, (Group C), and the Romanian clone, FRMA (Group F). The frequency of reactivity of the peptides from BRT3.6, CUG045, and FRMA were within the ranges of 57-100%, 50-100%, and 57-100%, respectively, for the sera collected from these disparate localities. In contrast to these findings, the V3 peptides derived from the other Ugandan isolates showed a more restricted pattern of reactivity with the same serum samples: AUG06c (1-63%), DUG23c (2%), and DUG044 (38-87%). The results from ELISA inhibition assay indicated that the V3 peptide from BRT 3.6, CUG045, and FRMA express closely related antigenic specificities quite distinct from those in AUG06c and DUG044. The residues comprising the PND in BRT 3.6, CUG045, and FRMA appear to be well conserved in the HIV-1 subtypes prevalent in the selected Ugandan locales.

PIP: In Uganda, health workers collected serum samples from 96 asymptomatic HIV-1 infected blood donors in Ishaka and Mbarara (southwest), Kisenyi and Kampala (central), and Lugazi and Jinja (east) so researchers working in a biochemistry laboratory at The City University of New York could describe the relative reactivity of the V3 loop from HIV-1 subtypes A, B, C, D, and F, as well as study the antigenic relationships within the PND encoded in the divergent HIV-1 subtypes. Regardless of geographic origin, the V3 peptides from most of the sera (at least 50%) collected in Uganda cross-reacted at high frequencies with the peptides derived from the novel North American clone (BRT3.6), the Ugandan clone (CUG045), and the Romanian clone (FRMA). The frequency of reactivity of these peptides with the test sera ranged from 57% to 100% for BRT3.6, from 50% to 100% for CUG045, and from 57% to 100% for FRMA. The V3 peptides from other Ugandan isolates (AUG06c, DUG044, and DUG23c) were less reactive with the same serum samples than BRT3.6, CUG045, and FRMA: 1-63%, 38-87%, and 2%, respectively. This finding suggests that the antigenic determinants expressed in AUG06c, DUG044, and DUG23c may not represent the PND encoded in most HIV-1 strains afflicting the Ugandan communities. The V3 peptides from BRT3.6, CUG045, and FRMA express closely related antigenic specificities altogether different from those in AUG06c and DUG044. The HIV-1 subtypes present in the selected Ugandan sites appear to effectively conserve the residues making up the PND in BRT3.6, CUG045, and FRMA.

Microcomputer system for automatic identification of the Cryptococcus neoformans and its clinical application.

In this study, microcomputer image processing and pattern recognition technology, and the knowledge of morphology and optical characteristics of Cryptococcus neoformans were used for identification of Cryptococcus neoformans. Four groups of mice were lethally infected with standard strain, Wuhan strain, American B-2643 strain and Var. Shanghainesis of the Cryptococcus neoformans. The samples collected included mice brain, lung, kidney, liver, small intestine tissue and were observed under a light microscope. More than 600 images of the fungus were input into a microcomputer. A system of computer for automatic identification of the Cryptococcus neoformans was developed. The technique involved image preprocessing, image segmenting, coding of line-length on the edge, curve fitting, extracting of image feature, building of image library and feature data bank etc.. And then, 768 images of the clinical samples and other fungus samples whose morphological features tend to be confused with Cryptococcus neoformans were input into microcomputer and subjected to automatic identification. The Cryptococcus neoformans was accurately identified within 15 min, and the consistency rate with results of routine culture was 98%.

Effects of 2-camphanone on canine portal vein blood flow and rat smooth muscle.

2-Camphanone is under clinical evaluation for alleviation of hemorrhoidal bleeding and inflammation. Reduced portal venous blood flow may distend, whereas improved portal venous blood flow may alleviate, hemorrhoidal vein distention. The effects of 2-camphanone on canine portal venous blood flow were investigated using pulsed Doppler flow techniques and on the spontaneous contractions of the isolated rat portal vein. Both intravenous (0.06, 0.2, and 0.6 mg/kg) and transdermal (6 mg/dog on the thigh) administration of 2-camphanone to dogs anesthetized with pentobarbital sodium increased portal venous flow velocity by 20%-30% without affecting femoral arterial blood flow, heart rate, or arterial blood pressure compared with vehicle-treated animals. Transdermal administration of 0.6, 2, and 6 mg of 2-camphanone, in a volume of 0.1 mL, to rats decreased the spontaneous contractions of the isolated rat portal vein in vitro. The data suggest that 2-camphanone exhibits a relatively selective effect on portal venous smooth muscle to reduce venous congestion and increase blood flow velocity. 2-Camphanone may be useful in the treatment not only of hemorrhoids, but also of esophageal reflux and portal hypertension.

Rotavirus infection of the oropharynx and respiratory tract in young children.

Oropharyngeal aspirates were obtained from 89 infants hospitalized with respiratory illnesses accompanied or not by diarrhea and 33 control patients without the diseases. Rotavirus was detected from 25 of these patients by immunocytology, isolation of the virus in cultures of MA104 cells, or both. None of the control patients gave a positive result. The infection involves squamous cells and globlet cells probably originating from the oropharynx, and ciliated columnar epithelial cells from the respiratory tract. The virus from 2 specimens was propagated by repeatedly passaging in the cultures and found to have characteristic morphology of rotavirus. The electrophoretic patterns of the viral RNA extracted from them are closely similar to those obtained with the rotavirus genome extracted from the stool of the same patients. Repeated stool specimens were also obtained, and sera were paired from some of these subjects. All but one of the patients who gave a positive virology for their aspirates also showed a significant rise in the titres of common group A rotavirus antibody, neutralizing antibody against one or more of serotypes of rotavirus, or both. Patients who excreted rotavirus in their stools were younger and had significantly lower titres of rotavirus antibodies in their acute sera, than those who shedded the virus in the oropharynx but did not excrete the virus in repeated stool specimens. The prevalence of rotavirus in the oropharyngeal aspirates from these patients surpassed that of adenovirus, respiratory syncytial virus, influenza virus, and herpes simplex virus combined.

SKF-525A does not inhibit release of endothelium-derived relaxing factor from rat thoracic aorta and dog mesenteric and femoral artery.

SKF-525A (proadifen) inhibits endothelium-dependent relaxations induced by acetylcholine, arachidonic acid and the calcium ionophore A23187. This suggests that SKF-525A is an inhibitor of endothelium-derived relaxing factor (EDRF) and that EDRF may be a product of arachidonic acid metabolism formed via a cytochrome P-450-dependent pathway or that EDRF release is dependent on cytochrome P-450. We tested this postulate using both isolated rings of rat thoracic aorta and dog mesenteric and femoral artery and the perfusion-superfusion bioassay. Rings of rat thoracic aorta and dog mesenteric and femoral artery with intact endothelium were precontracted with an EC50 concentration of norepinephrine (0.1 nmol/l) or U46619 (0.05 mumol/l) and the relaxation to acetylcholine (ACh), bradykinin, adenosine triphosphate (ATP) or nitroglycerin (GTN) were obtained before, 30 min after addition of, and 30 min after washout of SKF-525A (50 mumol/l). SKF-525A inhibited ACh-induced endothelium-dependent relaxation of rat aortic rings and endothelium-dependent relaxation of the dog mesenteric and femoral artery produced by ACh and ATP, but did not affect relaxation to bradykinin or GTN. The inhibitory effect on SKF-525A on ACh and ATP-induced relaxation was partially reversed upon its washout from the muscle chamber. Pretreatment of the blood vessels with ibuprofen (1 mumol/l) did not attenuate SKF-525A-mediated inhibition of the relaxations to any agonist. Selective exposure of dog femoral artery (donor) to SKF-525A (50 mumol/l) for 60 min did not affect the relaxation responses of endothelium-rubbed coronary artery (bioassay tissue) to basal EDRF nor to the effluent from donor tissues stimulated with ACh (10-1,000 pmol), ATP (1-100 nmol) or bradykinin (3-100 pmol). The results show that SKF-525A exhibited a reversible inhibition of endothelium-dependent relaxation by a smooth muscle mechanism unrelated to the generation of EDRF from endothelium.

[Studies on the chemical constituents of Excoecaria cochinchinensis Lour. var. viridis Merr].

Eight compounds have been obtained from the root and stem of Excoecaria cochinchinensis var. viridis growing in Tonghai county of Yunnan province. According to their spectroscopic analyses and physicochemical constants, they have been identified as: shikimic acid, 1-cyclohexene-1-carboxylic acid-5-hydroxy-3,4-isopropylidene-dioxy, oxy-bis(5-methylene-2-furaldehyde), beta-sitosterol, tetracosanoic acid, palmic acid, steric acid and hentriacontane.