Expression of a bioactive bovine interleukin-12 using baculovirus.

Recombinant baculoviruses that express recombinant bovine interleukin-12 (rboIL-12) subunits, p35 and p40 subunits were constructed. A recombinant virus containing the p40 subunit gene expressed the p40 subunit as a 40kDa monomer and an 80kDa disulfide-linked homodimer in the infected insect cells and in the culture supernatant. The p35 subunit was expressed in a 30kDa monomer in the infected cells but not in the supernatant. Superinfection of both recombinant viruses into the cells in a spinner flask resulted in the formation of a 70kDa disulfide-bonded heterodimer detected in the supernatant by immunoblotting using anti-p40 and anti-p35 subunits antibodies. The superinfected culture supernatant showed induction of IFNgamma mRNA synthesis and IFNgamma production in bovine peripheral blood mononuclear cells. Thus, the bioactive rboIL-12 was produced in large scale using a baculovirus expression system.

A case of postpregnancy osteoporosis.

A puerperant woman, who was previously healthy and had no disease known to affect bone metabolism, experienced lower back pain and lumbar vertebral fractures during lactation. Both bone formation markers and resorption markers were markedly elevated. Bone mineral density of the lumbar spine as measured by dual energy X-ray absorptiometry was extremely low. She stopped lactation through the use of bromocriptine because of the large volume of milk secretion. After treatment with calcitonin injections and the use of a corset, her back pain gradually disappeared. This case appears to be postpregnancy osteoporosis.

Comparison of bone metabolic markers between maternal and cord blood.

We studied 41 normal pregnant women and their neonates in order to compare bone metabolism between them. We examined more specific bone formation markers (intact osteocalcin, bone-specific alkaline phosphatase) and a recently developed and more sensitive bone resorption marker (C-telopeptide of type I collagen; CTX) than previously available in maternal and umbilical cord venous blood taken at delivery. The concentrations of all markers of bone turnover, including CTX, in cord serum were significantly higher than those in maternal serum. There was no significant correlation between maternal and cord serum levels for any marker. These results indicate that fetal bone turnover is markedly enhanced compared with maternal bone turnover and is independent of maternal bone metabolism in late pregnancy.

Changes in urinary excretions of C-telopeptide and cross-linked N-telopeptide of type I collagen during pregnancy and puerperium.

Previously we reported an increase in bone resorption during pregnancy and lactation by measuring pyridinoline (Pyr) and deoxypyridinoline (D-Pyr). To further assess bone metabolism during peripuerperal periods, we measured the urinary excretions of C-telopeptide (CTX) and cross-linked N-telopeptide (NTX) of type I collagen, new markers of bone resorption. In addition to Pyr and D-Pyr, urinary CTX and NTX were measured by two ELISAs which recognize the corresponding peptide of type I collagen after urine samples were collected cross-sectionally from 230 women who consisted of 187 pregnants, 25 puerperants, and 18 age-matched nonpregnant women. Urinary CTX was also measured longitudinally from 10 pregnants at 5-9, 28-31 and 36-39 weeks of gestation and 1, 3, 6 months after parturition. Similar to the changes in Pyr and D-Pyr, the mean CTX and NTX values significantly increased in the 3rd trimester of pregnancy and remained high during puerperium compared with nonpregnant or early pregnant women (P<0.05). In a longitudinal study, the mean CTX value significantly increased in the 3rd trimester of pregnancy and at 1 month of puerperium compared with that in the early stage of pregnancy (P<0.05). These results further confirm our previous evidence that bone resorption is enhanced during the 3rd trimester of gestation and puerperium and suggest that urinary CTX and NTX measured by ELISA, which is more convenient than HPLC, are useful markers to assess bone resorption during peripuerperal periods.

Changes in bone mass as determined by ultrasound and biochemical markers of bone turnover during pregnancy and puerperium: a longitudinal study.

In a longitudinal study, we analyzed the speed of sound (SOS) and broadband ultrasound attenuation (BUA) of the os calcis as an index of bone mineral density (BMD) to define the effects of pregnancy and lactation on bone metabolism. We used an ultrasound bone densitometer and measured 6 biochemical markers of bone turnover in 18 healthy women throughout pregnancy and puerperium. The measurement of SOS and BUA by such an ultrasound device was clinically advantageous; not only is it radiation-free technology, but it also correlates highly with BMD measured by conventional X-ray bone densitometry. While a significant decrease in SOS was found in the 3rd trimester of pregnancy as compared with the early stage of pregnancy, there was no difference in both SOS and BUA between the breast-feeding women and the principally formula-feeding women during a 6-month period of puerperium. The analysis of biochemical markers revealed that both bone formation and bone resorption were elevated in the 3rd trimester of pregnancy as well as during puerperium, and that the breast-feeding women had significantly higher bone metabolism than the principally formula-feeding women. These results indicate that bone mass decreases as bone turnover itself is enhanced during pregnancy, while lactation does not substantially affect bone mass during at least 6 months of puerperium, although bone turnover is active.

Neutrophil activation after percutaneous transluminal coronary angioplasty.

We investigated whether percutaneous transluminal coronary angioplasty (PTCA) would induce neutrophil activation in patients with coronary artery disease. Blood samples were taken from the coronary sinus in 14 patients who underwent PTCA and in 9 control subjects who underwent coronary arteriography (CAG). Flow cytometry was used to measure membrane surface expression of beta 2 integrin (CD11b) and the generation of hydrogen peroxide in neutrophils after ex vivo phorbol myristate acetate stimulation by 2,'7'-dichlorofluorescein. Neutrophil elastase was measured by an immunoenzymatic method. Surface expression of CD11b increased significantly, approximately twofold, after PTCA but not after CAG. Mean fluorescence intensity of 2',7'-dichlorofluorescein in stimulated neutrophils decreased significantly after PTCA, suggesting a previous in vivo activation, but not after CAG. Neutrophil elastase increased significantly after PTCA but not after CAG. These data indicate that PTCA induces neutrophil activation and suggest that neutrophils may contribute to the ischemic injury.

New noninvasive assessment of regional left ventricular function by digital subtraction angiography without the use of contrast medium.

We have developed a new noninvasive method to evaluate regional left ventricular (LV) function by digital subtraction angiography (DSA) without the use of contrast medium. DSA images of the left ventricle with and without contrast medium were obtained from 35 patients with anterior myocardial infarction (MI) and from 35 control subjects. Using an image-processing computer, regional LV time-density curves were constructed for one cardiac cycle. Regional LV time-density curves obtained from DSA without the use of contrast medium presented a pattern similar to those from intravenous DSA. The amplitude of regional LV time-density curves in patients with MI decreased along with increasing severity of regional wall motion abnormality assessed by conventional left ventriculography. In attempting semi-quantitative evaluation by DSA without the use of contrast medium, the regional wall motion index (RWI) in the 6 segments of the left ventricle was calculated by normalizing segmental density changes to the maximal segmental density changes. When compared with control subjects, patients with MI have significantly lower RWIs in the anterolateral and apical regions. RWI showed a good correlation with the regional ejection fraction (REF) obtained from intravenous contrast DSA (r = 0.83). RWI decreased with increasing severity of regional wall motion abnormality by qualitative analysis in conventional left ventriculography, being consistent with REF. The diagnostic accuracy of RWI therefore seemed to be comparable to that of REF derived from intravenous contrast DSA. These results indicate that computerized analysis of DSA without the use of contrast medium is a valuable noninvasive method for semi-quantitative assessment of regional LV function.

Relationship between coronary risk factor and arteriographic feature of coronary atherosclerosis.

Coronary arteriography was performed in 1,029 consecutive patients with ischemic heart disease and the relationship between the arteriographic features of coronary atherosclerosis and coronary risk factors was analyzed by case control studies. Patients were divided into four groups according to coronary arteriographic findings. Patients with normal or near normal coronary arteriograms (Group I) showed a high prevalence of smoking habit and a higher value of serum uric acid compared with the control group, so smoking and hyperuricemia were considered to be the risk factors for coronary atherosclerosis in patients of group. Four selected variables: smoking, hyperuricemia, hypertension and hyperlipidemia, were identified to be risk factors for the patients with minor plaques in the coronary arteries (Group II). As in Group I, smoking and hyperuricemia had a close relationship to solitary tight plaque in a branch of the coronary artery (Group III). Multiple tight stenoses in the coronary arteries (Group IV) correlated closely with smoking, hyperuricemia, hypertension, hyperlipidemia and diabetes mellitus. Thus, there were many strong risk factors for patients with diffuse, extended coronary atherosclerosis (Group II and Group IV), while only two factors, smoking and hyperuricemia, were considered to be risk factors for the patients with near normal coronary arteries ies or a solitary plaque in a branch of the coronary artery. These findings suggest that the role of the coronary risk factors on the pathogenesis of coronary atherosclerosis is not uniform but variable depending on the morphologic variability of the coronary atherosclerosis and on the pathophysiology of the ischemic heart disease.

[Pharmacokinetic and clinical studies of cefuzonam in obstetric and gynecologic infections].

Cefuzonam (CZON, L-105), a new semisynthetic cephem antibiotic, was studied in the field of obstetrics and gynecology, pharmacokinetically and clinically. Following is a summary of the results. 1. Concentrations of CZON in serum and genital organs following intravenous drip infusion of 1 g of the drug over 60 minutes were measured. Samples were taken during 40 to 190 minutes after the end of the infusion. Ratios of concentrations of the drug transferred into genital organs to the concentration in the cubital venous serum were 0.5 to 1.5 in many cases. Levels of the drug in the genital organs examined well exceeded MICs for common causative organisms in obstetric and gynecologic infections. 2. Therapeutic effects of 1 g or 2 g of CZON by drip infusion twice daily were evaluated in 8 patients. Overall clinical efficacy was 62.5% (5/8). No side effects or abnormal laboratory findings due to the drug were noted. The results suggest that CZON is a useful antibiotic for obstetric and gynecologic infections.