Intra-amniotic infection and premature rupture of the membranes

Antiphospholid antibodies (APL) have a notable association with recurrent miscarriages, arterial and venous thrombosis and thrombocytopenia. Analysis of the potential pathogenic effects of such human antibodies has been hampered by the considerable difficulty in producing IgG as opposed to IgM monoclonal immunoglobulins. We have developed four human monoclonal IgG APL (LJ1, AH2, DA3 and UK4) by fusing the peripheral blood lymphocytes of three patients with SLE with a mouse human heteromyeloma cell line, CB-F7. These antibodies bind to a variety of anionic phospholipids, two (LJ1 and AH2) bind total histones but none binds to ssDNA or dsDNA. Binding to beta 2 GPI is non-specific. UK4 alone demonstrates lupus anticoagulant activity. All four have lambda light chains, two are IgG1 (AH2 and UK4) and two are IgG3 (LJ1 and DA3). These APL utilize VH genes present in the fetally restricted repertoire and multiple somatic mutations in the CDR suggest an antigen-driven process. In contrast, there is no restriction in V lambda gene usage and only one lambda chain is extensively mutated. Two clonally related hybridomas were isolated from a single patients. This supports the theory that clonal expansion is the mechanism whereby antigen selects high affinity mutations.     

Clinical evaluation--continuous real-time intra-arterial blood gas monitoring during anesthesia and surgery by fiber optic sensor

A clinical evaluation of the clinical utility, techniques of use, durability, accuracy, and potential complications of a newly available system for the continuous real-time, intra-arterial monitoring of arterial blood gas and acid base status (ABG) has been studied (Optex BioSentry System, Optex Biomedical, Incorporated, The Woodlands, Texas, U.S.A.). The system consists of three separate fiber optic channels with contained fluorescent and absorbant chemical analytes imbedded in a single probe and capable of insertion inside of a twenty gauge indwelling arterial catheter (The Optex Optode Sensor), along with an external monitor. The system was utilized during anesthesia and surgery in the care of five informed patients. Constant trend monitoring of all three variables was deemed satisfactory in four of the patients. The fifth sensor was damaged by a surgical assistant while in place and ceased to function. Comparison of Optode sensor readings with standard clinical laboratory ABG analysis was excellent in three uncomplicated patients (pH: bias -0.0183 pH units, precision: 0.0237 pH units; PCO2: bias 3.22 mmHg, precision 2.04 mmHg; PO2: bias -5.94 mmHg, precision 11.74 mmHg). Postoperative study suggested that discrepancies in a fourth patient may have been due to an incorrect 'offset' applied to the Optode sensor yielding a constant error.     

Differences in lipoprotein lipid concentration and composition modify the plasma distribution of cyclosporine

PURPOSE: The purpose of this study was to define the relationship between lipoprotein (LP) lipid concentration and composition and the distribution of cyclosporine (CSA) in human plasma. METHODS: 3H-CSA LP distribution was determined in normolipidemic human plasma that had been separated into different LP and lipoprotein-deficient plasma (LPDP) fractions by either affinity chromatography coupled with ultracentrifugation, density gradient ultracentrifugation or fast protein liquid chromatography. 3H-CSA LP distribution (at a concentration of 1000 ng/ml) was also determined in patient plasma samples with defined dyslipidemias. Furthermore, 3H-CSA LP distribution was determined in patient plasma samples of varying LP lipid concentrations. Following incubation, the plasma samples were separated into their LP and LPDP fractions by sequential phosphotungistic acid precipitation in the dyslipidemia studies and by density gradient ultracentrifugation in the specific lipid profile studies and assayed for CSA by radioactivity. Total plasma and lipoprotein cholesterol (TC), triglyceride (TG) and protein (TP) concentrations in each sample were determined by enzymatic assays. RESULTS: When the LP distribution of CSA was determined using three different LP separation techniques, the percent of CSA recovered in the LP-rich fraction was greater than 90% and the LP binding profiles were similar with most of the drug bound to plasma high-density (HDL) and low-density (LDL) lipoproteins. When 3H-CSA was incubated in dyslipidemic human plasma or specific patient plasma of varying LP lipid concentrations the following relationships were observed. As the very low-density (VLDL) and LDL cholesterol and triglyceride concentrations increased, the percent of CSA recovered within the VLDL and LDL fractions increased. The percent of CSA recovered within the HDL fraction significantly decreased as HDL triglyceride concentrations increased. The percent of CSA recovered in the LPDP fraction remained constant except in hypercholesterolemic/hypertriglyceridemic plasma where the percent of CSA recovered decreased. Furthermore, increases in VLDL and HDL TG/TC ratio resulted in a greater percentage of CSA recovered in VLDL but less in HDL. CONCLUSIONS: These findings suggest that changes in the total and plasma LP lipid concentration and composition influence the LP binding of CSA and may explain differences in the pharmacological activity and toxicity of CSA when administered to patients with different lipid profiles.     

Is fecundability associated with month of birth? An analysis of 19th and early 20th century family reconstitution data from The Netherlands

The relationship between fecundability and month of birth was investigated in a cohort of 1526 women who married between 1802 and 1929, using only women whose first marriage occurred before the age of 35 years. On the basis of their time to pregnancy (TTP, calculated as time between wedding and first birth minus gestational length), women were categorized into two groups: fecunds (TTP up to 12 months or prenuptial conceptions, n = 1348) and subfecunds (TTP >18 months, n = 118). By use of logistic regression, cosinor functions with a period of 1 year or 6 months and variable shift and amplitude were fitted through the monthly odds of subfecunds versus fecunds. The best fitting curve was unimodal, with a zenith in September (P = 0.13 for H0: no differences). Exclusion of childless women (n = 36, minimum follow-up 5 years) from the subfecunds led to a similar curve (P < 0.01), while childless women, as compared with fecunds, showed a birth distribution that was best represented with a bimodal curve with zeniths in January and July (P = 0.06). This study provides evidence for the existence of differences in fecundability by month of birth. The cause of this relationship is unclear, but may lie in a melatonin-dependent circannual variability of the quality of the oocyte.     

Fos and serotonin immunoreactivity in the raphe nuclei of the cat during carbachol-induced active sleep: a double-labeling study

The microinjection of carbachol into the nucleus pontis oralis produces a state which is polygraphically and behaviorally similar to active sleep (rapid eye movement sleep). In the present study, using double-labeling techniques for serotonin and the protein product of c-fos (Fos), we sought to examine whether immunocytochemically identified serotonergic neurons of the raphe nuclei of the cat were activated, as indicated by their expression of c-fos, during this pharmacologically-induced behavioral state (active sleep-carbachol). Compared with control cats, which were injected with saline, active sleep-carbachol cats exhibited a significantly greater number of c-fos-expressing neurons in the raphe dorsalis, magnus and pallidus. Whereas most of the c-fos-expressing neurons in the raphe dorsalis were small, those in the raphe magnus were medium-sized and in the raphe pallidus they were small and medium-sized. The mean number of serotonergic neurons that expressed c-fos (i.e. double-labeled cells) was similar in control and active sleep-carbachol cats. These data indicate that there is an increased number of non-serotonergic, c-fos-expressing neurons in the raphe dorsalis, magnus and pallidus during the carbachol-induced state.(ABSTRACT TRUNCATED AT 250 WORDS)     

Mapping and characterization of novel (CAG)n repeat cDNAs from adult human brain derived by the oligo capture method

BACKGROUND: The role of chest CT scan in the assessment of patients with hemoptysis is uncertain. AIM: To evaluate the usefulness of CT scan in patients with non massive hemoptysis. PATIENTS AND METHODS: Ninety six patients, 60 male, aged 23 to 76 years old, who presented with hemoptysis to an University Hospital, were studied. All patients were studied with a chest radiograph, a fiberoptic bronchoscopy and a high resolution CT scan. RESULTS: The final causes of hemoptysis were bronchiectasis in 27 cases, bronchogenic carcinoma in 24 cases and lung infections in nine. The source of bleeding was not identified in 18 patients (19%). CT scan clarified abnormalities seen in the chest radiograph in 30 patients (31%) and provided new diagnostic information in 13 (14%). CT scan correctly localized the source of bleeding found by fiberoptic bronchoscopy in 35 of 43 patients (81%), whereas chest radiograph did so in 27 (77%). All patients with bronchogenic carcinoma were identified by chest radiograph or bronchoscopy. Twenty of the 27 patients with bronchiectasis had radiological abnormalities in the chest radiograph. In only two patients, with lung metastases and non conclusive chest radiograph and bronchoscopy, CT scan provided information that significantly modified clinical management. CONCLUSIONS: CT scan was useful to stage patients with bronchogenic carcinoma and to assess the extension of bronchiectasis, but its impact in the management and clinical evolution of patients was limited. Therefore we do not recommend the routine use of CT scan in the assessment of patients with hemoptysis.     

Analysis of lipidation of recombinant Lyme vaccine protein (rOspA) by electrospray mass spectroscopy

Lipidation at the N-terminal of recombinant outer surface protein A (rOspA) is an important determinant of its immunogenicity. Lipidation patterns of rOspA can be sensitive to processing environments and storage conditions. In order to assure product consistency and stability. it is essential to characterize and monitor lipidation patterns of rOspA through its life-cycle. Electrospray mass spectroscopy combined with maximum entropy calculation was employed to analyze the lipidation of rOspA. The results revealed that more than 90% of protein is a tri-lipidated rOspA and the remainder is di-lipidated. It was demonstrated that the method is both sensitive and quantitative and has the potential to be used for routine quality control and stability testing.     

Structural analyses of gp45 sliding clamp interactions during assembly of the bacteriophage T4 DNA polymerase holoenzyme. II. The Gp44/62 clamp loader interacts with a single defined face of the sliding clamp ring

In the preceding paper (Latham, G. J., Bacheller, D. J., Pietroni, P. , and von Hippel, P. H. (1997) J. Biol. Chem. 272, 31677-31684), we demonstrated that the T4 gp44/62-ATP clamp loader binds to the C-terminal face of the gp45 sliding clamp. Here we extend these results by exploring the structural relationship between the gp43 polymerase and the gp45 sliding clamp. Using fluorescence intensity and polarization techniques, as well as photo-cross-linking methods, we present evidence that gp43, like gp44/62, binds to the C-terminal face of gp45. In addition, we show that g43 binds to the gp45 clamp in two distinct interaction modes, depending on the presence or absence of template-primer DNA. When template-primer DNA is present, gp43 binds tightly to gp45 to form the highly processive DNA polymerase holoenzyme. Gp43 also binds to gp45 in the absence of template-primer DNA, but this interaction is more than 100 times weaker than gp43-gp45 binding on DNA. Specific interactions between gp43 and the C-terminal face of gp45 are maintained in both modes of binding. These results underscore the pivotal role of template-primer DNA in modulating the strength of protein-protein interactions during DNA synthesis and provide additional insight into the structural requirements of the replication process.