Analysis and purification of phorbol esters using normal phase HPLC and photodiode-array detection

For the first time a normal-phase HPLC method using photodiode-array detection is described for the analysis and purification of phorbol esters. The use of the method is demonstrated with examples of 10 different tigliane and daphnane esters (TPA, DOPP, DOPPA, Sap A, Sap B, Sap C, Sap D, Thy A, Ro and Rx). Both analytical and semipreparative techniques were developed. The method has been used in the final purification of DOPP and Rx from plant extracts. The method can be employed in the areas of phytochemistry, biochemistry and pharmacology/toxicology, where small samples of the toxic materials are required for research.     

A pilot study of the marginal adaptation and surface morphology of glass-cermet cements

This study investigated changes in the marginal adaptation and surface morphology of Ketac-Silver and Chelon-Silver glass-current cements over time. Dispersalloy amalgam was used as a control. Contralateral pairs of carious primary molars were restored with the test materials and amalgam. Clinical evaluations were scheduled at 12, 18, and 24 months after placement. Gold-plated replicas of the restorations were observed with scanning electron microscopy. Fractures and cracks in the surface of the Dispersalloy and Chelon-Silver increased the surface roughness; however, the damage was superficial and self-limiting in the Dispersalloy restorations, while in Chelon-Silver the fractures caused the material to break down in layers. A substantial quantity of pores, usually smaller than 50 microns in diameter, were observed throughout the surface of the Chelon-Silver restorations. The pores in the surface of Ketac-Silver were fewer and smaller. The incidence of cavomarginal breakdown increased with time. Chelon-Silver restorations had a higher rate of cavomarginal breakdown than did Ketac-Silver and Dispersalloy restorations up to 18 months. However, there was no statistically significant difference in the marginal adaptation of the three groups at 24 months.     

The immunomodulating action of cyclosporin A in vitro and in vivo on the lymphocytes of patients with alveolitis

The effect of various concentrations of ofloxacin, ciprofloxacin and lomefloxacin on the morphokinetic parameters of the cellular elements of the lung tissue of intact animals (mice, guinea pigs and dogs) was studied under the conditions of the tissue culture. It was shown that in a concentration of 100 micrograms/ml and the exposure time of 24 hours the drugs had no effect on the mobility and structure of the lung tissue cellular elements. When the drugs were used in a concentration of 100 micrograms/ml the mobility rate of the lymphocytes and macrophages proved to by markedly retarded by the end of the 24th hour. In a concentration of 1000 micrograms/ml the drugs killed the cell culture. There were detected no specific differences in the effect of the fluoroquinolones on the cellular elements of the lung tissue of the intact animals.     

Agreement and clinical utility of 2 techniques for measuring cardiac output in patients with low cardiac output

BACKGROUND: The reliability of cardiac output obtained with the bolus technique is a problem. OBJECTIVES: To compare measurements of cardiac output measured with bolus and continuous techniques in patients with low cardiac output and to determine if measurements obtained with the continuous technique increased the number of subsequent clinical decisions. METHODS: In 60 intensive care patients, a nurse recorded a single continuous cardiac output measurement and then obtained the mean of 3 consecutive bolus determinations. The medical records of these 60 patients (experimental group) for the next 48 hours and of 60 other patients with regular or mixed venous oximetry catheters (control group) were reviewed to assess the occurrence of cardiac output events and the frequency of clinical decisions based on the events. RESULTS: Mean cardiac output was 4.46 L/min by the continuous technique and 5.20 L/min by the bolus technique (P = .011) for the experimental group. Median bias between the 2 types of measurements was -0.10 L/min (P = .79). Twenty-three of the pairs (38%) had an absolute percent difference greater than 15%. Of these, 18 (78%) had a higher bolus reading. Treatment decisions per 48 hours were 9.9 for the experimental group and 8.6 for the control group (P = .014). Median length of stay was 2 days less in the experimental group (P = .02), and mean highest cardiac output was 0.81 L/min higher (P = .009). CONCLUSIONS: Measurements of cardiac output determined with the continuous technique may be more precise than measurements determined with the bolus technique. Continuous cardiac output information increases the number of treatment decisions and actions that may shorten hospital length of stay.     

Assessing neuromuscular block at the larynx: the effect of change in resting cuff pressure and a comparison with video imaging in anesthetized humans

Neuromuscular block (NMB) at the larynx has been assessed by measuring the cuff pressure (CP) in an endotracheal tube (ETT) placed between the vocal cords. In this study, we evaluated the decrease in resting cuff pressure (RCP) after the administration of rocuronium and the effect of this decrease on the assessment of NMB, and we compared CP measurement with an alternative technique, video imaging (VI). In 20 patients, NMB was determined at the hand by mechanomyography and at the larynx initially by CP and subsequently by VI, recording images using a fiberoptic bronchoscope via a laryngeal mask. Train-of-four stimuli were applied at both sites. After baseline measurements, the ETT was replaced, and rocuronium was infused to achieve a steady-state 50% (n = 10) or 75% (n = 10) block at the hand. CP measurements were recorded before and after restoration of RCP to prerocuronium pressure, followed by further VI measurements. The mean RCP decreased from 21 +/- 4 to 12 +/- 5 mm Hg after rocuronium. At 50% block at the hand, the CP estimate of block at the larynx with reduced RCP was 62% +/- 18%, and that after restoring RCP was 29% +/- 13%; VI estimated 27% +/- 14% block. At 75% block at the hand, CP and VI estimated 52% +/- 11% and 46% +/- 9% block, respectively (RCP maintained). We conclude that RCP decreases after the administration of rocuronium, that restoring RCP significantly alters CP estimates of NMB, and that VI is in agreement with CP measurement if RCP is maintained at prerelaxant values. Implications: In this study, we show that a muscle relaxant-induced decrease in resting tension at the larynx may confound the assessment of neuromuscular block by cuff pressure measurement. The preliminary data suggest that video imaging may provide a suitable alternative to cuff pressure measurement to assess neuromuscular block at the larynx.     

PARTICLE STIMULATED NUCLEATION AND THE FORMATION OF RSCRYSTALLIZATION TEXTURE IN Al-Mn ALLOY CONTAINING PARTICLES Ⅱ. Interaction Between Particles and Other Nucleation Sites and the Formation of Recrystallization Texture

本文报了Ａｌ－Ｍｎ合金形昧形变取向基体中粒子周围亚晶转动规律及对应的粒子促进形核晶粒的取向特征的基础上,进一步测定了处在晶界,立方带上的了周转新晶粒的取向分布。     

Role of paired basic residues in the expression of active recombinant galactosyltransferases from the bacterial pathogen Neisseria meningitidis

The lgtB gene encoding a beta-1,4-galactosyltransferase gene and the lgtC gene encoding an alpha-1,4-galactosyltransferase from the bacterial pathogen Neisseria meningitidis were cloned into an expression vector and overexpressed in Escherichia coli. Both genes expressed very well, but problems with C-terminal proteolysis were encountered with both proteins. The lgtC protein was initially isolated from extracts of recombinant E.coli as a truncated species that retained enzymatic activity, and was subsequently shown by mass spectrometry to be 19 residues shorter than the expected protein. A specific set of engineered C-terminal deletions was constructed to investigate their effect on the expression of lgtC. As many as 28 residues could be deleted with little effect on activity, and with the concomitant improvement of the overall expression up to fivefold over the full length protein. The lgtB protein was also proteolysed in extracts of normal E.coli strains into enzymatically inactive fragments lacking 28 or 41 C-terminal residues. This degradation could be prevented by expression in an ompT protease deficient strain of E.coli. The full length lgtB protein was not stable in soluble protein extracts from all recombinant strains, however a stable enzyme preparation could be achieved with the membrane fraction from cells of the ompT deficient strain expressing lgtB. Specific deletions of lgtB were also constructed, and 15 residues could be removed without loss of enzyme activity and also with the concomitant improvement of the overall expression up to twofold over the full length protein. Longer deletions produced protein but activity could not be detected in these recombinant strains. Examination of the glycosyltransferase sequences from a wide range of bacteria showed their C-terminal segments of approximately 50 amino acids frequently contained paired basic residues. Engineering of these segments may therefore be required as a general practice to produce these enzymes for use in the large scale chemi-enzymatic synthesis of carbohydrate-based therapeutics.      

IFN-tau: a novel subtype I IFN1. Structural characteristics, non-ubiquitous expression, structure-function relationships, a pregnancy hormonal embryonic signal and cross-species therapeutic potentialities

IFN-tau (IFN-tau) constitutes a new class of type I IFN which is not virus-inducible, unlike IFN-alpha and IFN-beta, but is constitutively produced by the trophectoderm of the ruminant conceptus during a very short period in early pregnancy. It plays a pivotal role in the mechanisms of maternal recognition of pregnancy in ruminants and it displays high antiviral and antiproliferative activities across species with a prominent lack of cytotoxicity at high concentrations in vitro in cell culture and possibly in vivo. It exhibits high antiretroviral activity against HIV and exhibits immunosuppressive activity in a multiple sclerosis model and reduces embryo and fetal mortality by stimulation of IL-10 production. In this review all the biochemical and para-hormonal properties of this novel IFN-tau are described in detail: structural characteristics of proteins and genes, trophoblast expression, regulation of its expression, structure of its gene promoter, its absence in human species and in non-ruminant animals, the evolution of the IFN-tau genes, its structure-function relationships with its three-dimensional structure, structural localization of biological activities, its lack of cytotoxicity and its receptor. Surprisingly, for an IFN, IFN-tau is also a pregnancy-embryonic signal with paracrine antiluteolytic activity. In order to maintain luteal progesterone secretion, IFN-tau inhibits PGF-2alpha pulsatile secretion and oxytocin uterine receptivity in early pregnancy. It is believed to suppress pulsatile release of endometrial PGF-2alpha by preventing oxytocin and estrogen receptor expression. Additionally, it directly regulates prostaglandin metabolism and possibly the PGE:PGF-2alpha ratio.     

Antibiotic therapy for seminal infection. Effect on antioxidant activity and T-helper cytokines

OBJECTIVE: To investigate the effect of antibiotic therapy on seminal infection. STUDY DESIGN: The seminal plasma of 50 men was evaluated in three groups: (1) men with seminal infection (20), (2) men with leukocytospermia only (18), and (3) men of proven fertility (12). The evaluation protocol included semen analysis, culture and antibiotic sensitivity test, total antioxidant activity, alpha-tocopherol and retinol, T-helper cytokines, IL-2, IL-8, IL-4 and antisperm antibodies. RESULTS: Sperm parameters were worse with seminal infection: 25 versus 84 million per milliliter for fertile men. Antioxidant activity, total alpha-tocopherol and retinol were reduced in leukocytospermia (P < .02, .01) and seminal infection (P < .01, .05) as compared to controls. Antisperm antibodies IL-2 and IL-8 were highly expressed, while IL-4 was low in men with leukocytospermia and bacteriospermia. Gram-negative organisms were more associated with expression of T-helper 1 cytokines than T-helper 2 cytokines. Antibiotic therapy significantly improved the sperm parameters, antioxidant activity and IL-4 but reduced IL-2 and IL-8 and had no effect on antisperm antibody titer. CONCLUSION: Antibiotic therapy improves sperm parameters by increasing antioxidant activity and IL-4 and by reducing IL-2 and IL-8.