Technical note: Use of a simplified equation for estimating glomerular filtration rate in beef cattle.

This study was performed to clarify whether a formula (Holstein equation) based on a single blood sample and the isotonic, nonionic, iodine contrast medium iodixanol in Holstein dairy cows can apply to the estimation of glomerular filtration rate (GFR) for beef cattle. To verify the application of iodixanol in beef cattle, instead of the standard tracer inulin, both agents were coadministered as a bolus intravenous injection to identical animals at doses of 10 mg of I/kg of BW and 30 mg/kg. Blood was collected 30, 60, 90, and 120 min after the injection, and the GFR was determined by the conventional multisample strategies. The GFR values from iodixanol were well consistent with those from inulin, and no effects of BW, age, or parity on GFR estimates were noted. However, the GFR in cattle weighing less than 300 kg, aged<1 yr old, largely fluctuated, presumably due to the rapid ruminal growth and dynamic changes in renal function at young adult ages. Using clinically healthy cattle and those with renal failure, the GFR values estimated from the Holstein equation were in good agreement with those by the multisample method using iodixanol (r=0.89, P=0.01). The results indicate that the simplified Holstein equation using iodixanol can be used for estimating the GFR of beef cattle in the same dose regimen as Holstein dairy cows, and provides a practical and ethical alternative.

Glomerular filtration rate in Holstein dairy cows estimated from a single blood sample using iodixanol.

The isotonic, nonionic, contrast medium iodixanol, as a test substance, was compared with the conventional glomerular filtration rate (GFR) tracer inulin to establish a simplified procedure for estimating the GFR in Holstein dairy cows. First, inulin and iodixanol were coadministered as a bolus intravenous injection to clinically healthy cows at 30 mg/kg and 10mg of I/kg of body weight, respectively, followed by blood collection for multisample strategies. Serum iodixanol and inulin concentrations were separately determined by using HPLC and colorimetry, respectively, and blood urea nitrogen and creatinine concentrations in sera were measured. In the multisample method, the GFR values estimated by iodixanol were consistent with those estimated by inulin. No effects of body weight, age, or parity on GFR estimates were noted with either protocol used. No difference was observed between the GFR values obtained from nonlactating and lactating cows, suggesting that no transfer of iodixanol to milk occurred. An equation for calculating the GFR in the single-sample method was derived from the injected dose, sampling time, serum concentration, and estimated volume of distribution based on data from the multisample method in clinically healthy cows and cows with reduced renal function. The GFR values estimated by the single-sample method were in good agreement with those calculated by using the multisample method. These results demonstrate that the single-sample method using iodixanol can be applied as an alternative procedure for screening GFR in dairy cows.

A single-blood-sample method using inulin for estimating feline glomerular filtration rate.

BACKGROUND: Application of a multisample method using inulin to estimate glomerular filtration rate (GFR) in cats is cumbersome. OBJECTIVES: To establish a simplified procedure to estimate GFR in cats, a single-blood-sample method using inulin was compared with a conventional 3-sample method. ANIMALS: Nine cats including 6 clinically healthy cats and 3 cats with spontaneous chronic kidney disease. METHODS: Retrospective study. Inulin was administered as an intravenous bolus at 50 mg/kg to cats, and blood was collected at 60, 90, and 120 minutes later for the 3-sample method. Serum inulin concentrations were colorimetrically determined by an autoanalyzer method. The GFR in the single-blood-sample method was calculated from the dose injected, serum concentration, sampling time, and estimated volume of distribution on the basis of the data of the 3-sample method. RESULTS: An excellent correlation was observed (r = 0.99, P = .0001) between GFR values estimated by the single-blood-sample and 3-sample methods. CONCLUSIONS AND CLINICAL IMPORTANCE: The single-blood-sample method using inulin provides a practicable and ethical alternative for estimating glomerular filtration rate in cats.

[Examination of soluble cell adhesion molecules in pregnancy induced hypertension].

This study was performed to determine whether or not the soluble-intercellular adhesion molecule-1 (sICAM-1), vascular cell adhesion molecule-1 (sVCAM-1) and endothelial leukocyte adhesion molecule-1 (sELAM-1) are sensitive markers of pregnancy induced hypertension (PIH). sICAM-1 concentrations were significantly higher (p < 0.05) in the mild PIH compared to non-pregnant women and normal pregnant groups. sVCAM-1 concentrations in the mild PIH group and the severe PIH group were significantly higher than the non-pregnant women group (p < 0.0001, p < 0.01, respectively) and the normal pregnant group (p < 0.0001, p < 0.0005, respectively). The concentrations of sELAM-1 in the mild PIH group were also significantly higher compared to normal pregnant group (p < 0.01). Our results suggest that soluble cell adhesion molecules may be useful markers detecting endothelial damage and dysfunction in patients with PIH.

Differences among acute, subacute, and chronic chorioamnionitis based on levels of inflammation-associated proteins in cord blood.

The serum concentration of inflammation-associated proteins and several complement components in the cord blood of 215 newborns with and without chorioamnionitis (CAM), who were delivered between 17 and 42 weeks of gestation, were measured. We investigated the relationship of levels of serum proteins to acute, subacute, and chronic CAM, and to subacute necrotizing funisitis (SNF). Complement components C3d, C3, and C4 levels increased in subacute CAM (P = 0. 0002, P = 0.0007, P = 0.0029, respectively), whereas factor B increased in each type of CAM (P = 0.0001, P = 0.0009, P = 0.0004, respectively). Among the immunoglobulins, IgG levels were unrelated to the presence or type of CAM, IgM levels increased in subacute CAM (P < 0.0001), and IgA levels increased in chronic CAM (P < 0.0001). Among the acute phase reactants (APR), haptoglobin and C-reactive protein (CRP) levels increased in acute (P < 0.0001, P = 0.0022, respectively) and chronic CAM (P = 0.0035, P = 0.0345, respectively), whereas orosomucoid levels increased in chronic CAM (P = 0.0003). IL-6 levels increased in acute (P = 0.0011) and subacute (P = 0. 0475) CAM. C3d (P = 0.0063), C3 (P = 0.0289), C4 (P = 0.0491), and IgM (P < 0.0001) levels were increased in SNF. These findings suggest that the histologic distinction of acute, subacute, and chronic CAM is a useful indicator of the inflammatory mediator status of the infants. The infants with SNF may have ended their initial active inflammatory states, but they still have subacute immune activation.

[A case of congenital complete heart block in a mother with anti-52 kD SS-A/Ro antibodies].

We report a case of congenital complete heart block (CCHB). A 38-year-old woman was admitted our hospital because of fetal bradycardia at 21 weeks 3 days of gestational age. She had no symptom of collagen disease. On admission, laboratory data showed positive anti-nuclear antibodies, anti-SS-A/Ro antibodies and anti-52 kD SS-A/Ro antibodies. But anti-60 kD SS-A/Ro and anti-SS-B/La antibodies were negative. Consequently anti-52 kD SS-A/Ro antibodies positive woman had an infant with CCHB. The baby was equipped with pacemaker at the age of 2 months. This report suggests that anti-52 kD SS-A/Ro antibodies may play an important role in the development of CCHB.

Effect of chorioamnionitis on the levels of serum proteins in the cord blood of premature infants.

OBJECTIVE: To determine the relationships among complements, other serum proteins (including acute-phase reactant), and the cytokine interleukin-6. DESIGN: Prospective observational study. SETTING: A perinatal center, children's hospital, and research institute in Osaka, Japan. PATIENTS: Two hundred fifteen newborn infants ranging from 17 to 42 weeks in gestational age. MAIN OUTCOME MEASURES: We measured the serum levels of several proteins and complements in the cord blood of neonates with and without chorioamnionitis by immunological assays. RESULTS: The levels of C3d, haptoglobin, interleukin-6, IgM, C-reactive protein, and IgA were not influenced by gestational age, and levels of C5, C1q, C2, albumin, C9, and IgG were not influenced by chorioamnionitis status. The levels of C3, C4, CH50, factor B, and orosomucoid were influenced by both factors. CONCLUSIONS: Our investigation of the mutual relation of the data suggests that the increase of cytokine interleukin-6 affects each other component. We suggest that, compared with the serum levels of proteins in neonates with chorioamnionitis, levels in neonates without chorioamnionitis are more similar to those in the normal fetus.

Significance of chorioamnionitis.

In preterm deliveries, we have reported a high incidence (30-50%) of histologic chorioamnionitis (CAM) in the placenta. There is little evidence about the effects of CAM on preterm infants. We investigated the levels of complements and cytokines in the cord blood, the pathological nature of the placenta, the L/S ratio of gastric and tracheal aspirate of each preterm infant at birth, and assessed the biological effects of CAM on them. CAM stimulates the immunological system by cytokine production (IL6 and IL8) and complement activation in the fetus. It has been suggested that CAM may be one of the factor accelerating fetal maturation of the immunological system such as complement activation and immunoglobulin production, and of surfactant synthesis in the lung. On the contrary, CAM may damage the structures along the lining cells in the airway by accumulating polymorphonuclear cells of the infants with Wilson-Mikity syndrome.

[Restriction endonuclease analysis of Chlamydia trachomatis DNA].

Twelve serovar type strains (C, D, E, F, G, H, I, J, K, L1, L2 and L3) and eight isolates (D, 4; E, 2; K, 2) of C. trachomatis were examined by restriction endonuclease analysis (REA) of DNA extracted from the elementary bodies. No difference was observed in DNA patterns of three serotypes (L1, L2 and L3) of the biovar LGV when they were digested with EcoRI and analysed by electrophoresis in a 0.6% agarose gel. The genital strains of the biovar trachoma (serovars D-K) showed similar EcoRI patterns with or without detectable differences. Serovar C of the biovar trachoma differed from the biovar LGV and the genital strains. Comparative analysis of DNAs digested with EcoRI, BamHI, HindIII, SalI and NcoI revealed that C. trachomatis isolates belonging to serovars D and K, but not E, could be subdivided into different genome types. These results suggest that DNA cleavage pattern analysis is useful for epidemiological, clinical, and taxonomic studies of C. trachomatis.

[Distribution of serovars of Chlamydia trachomatis isolates in Japan].

To study the serovar distribution of C. trachomatis in Japan, a total of 85 genital C. trachomatis isolates from male and female patients attending the clinics were examined by the microimmunofluorescence test using immune sera of the isolates produced in mice. Of these isolates, 34 (40.0%) were typed D or E, and 19 (22.4%) were typed G or F. The serovars of the remaining 32 isolates were B, H, I, J, and K, and the proportions of these serovars were from 8.2 to 3.5%. Thus, two thirds of C. trachomatis isolates in this country were found to fall into only four serovars, namely, D, E, G, and F, and, therefore, the epidemiology of C. trachomatis infection in Japan seems to be similar to that of other countries in North America and Europe. The relative distribution of serovars of C. trachomatis isolates from male patients and female patients somewhat differed. Serovars D, E, and G, F were isolated in the same ratio from male patients, while the isolation ratio of the former serovars was three times or more higher than the latter serovars in female patients. No isolate typed serovar K was found in male patients, while 15% of isolates from female patients were typed this serovar.

Complement function and the synthesis of lung surfactant may be a regulation which preterm infants have in common.

The levels of CH50, the complement components, C1q, C4, and C3, C3 degradation fragments, and factor B were determined in the cord blood of 128 newborn infants. The levels of C3, C4, and C3d3 (an index of chronic in vivo complement activation) were clearly lower in the 28 infants with respiratory distress syndrome (RDS) than in the infants with other lung diseases or with normal lungs. CH50 and factor B levels were also low in RDS. Levels of C1q and other serum components in RDS infants were similar to the average levels in other infants without RDS at a corresponding gestational age. Lung surfactant is synthesized in alveolar type 2 cells, in which the complement components C4, C3, and factor B, but not C1q, have been reported to be synthesized. It seems possible that common factors regulate the synthesis of some complement components and surfactant.

[Evaluation of in vitro activity of rokitamycin against Chlamydia trachomatis].

Evaluation of the in vitro activity of rokitamycin (RKM) against Chlamydia trachomatis in cycloheximide treated HeLa 229 cells and McCoy cells by comparing with five drugs including doxycycline (DOXY), minocycline (MINO), ofloxacin (OFLX), ampicillin (ABPC) and erythromycin (EM) with regard to assaying minimal inhibitory concentrations (MICs), minimal lethal concentrations (MLCs), and by yield reduction assays: 1) direct treatment of Chlamydial organisms with various concentrations of antibiotics before inoculation, 2) pre-treatment of host cell (HeLa 229) with the antibiotics before they are infected and 3) treatment of already infected cultures (48 hrs after infection) with antibiotics. The yield of Chlamydia was determined by both assaying the infectivity of Chlamydia and/or Chlamydiazyme value (from Abott Co Ltd USA). It was found that similar MIC was obtained among the drugs tested (except EM) in both HeLa 229 cell and McCoy cell assay system. The MLC of RKM (0.3 micrograms/ml) was the same as that of OFLX and was significantly lower than that of other drugs tested. When Chlamydial organisms and the host cells were treated with various concentrations (25-0.1 micrograms/ml) of the drug, the infectivity and the growth of Chlamydia was noteworthily decreased with RKM treatment. Infectivity of Chlamydia in an already infected cultures also decreased with RKM treatment within 24 hours when comparing the value of the control. In other drugs treatment, 96 hours or more hours were required for obtaining the same infectivity as RKM.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of amnionitis on the complement system of preterm infants.

The development of the complement system was studied by quantitation of total hemolytic complement activity (CH50), C1q, C3, C4, and C3 split product (C3d) in cord plasma of nine human fetuses (17-22 weeks of gestation), 110 preterm (24-36 weeks of gestation) and 30 term neonates. The complement levels were analyzed in relation to various illnesses of preterm infants. Histological examination of the placenta revealed a higher incidence of amnionitis in the placenta of less than 34 gestational weeks. In cases without amnionitis, there were significant correlations between complement levels and gestational age. In cases with amnionitis, the complement system was activated even in infants of less than 28 weeks gestation. The complement levels correlated with the extent of the inflammation in the placentas and umbilical cords except for C1q. In infants with Wilson-Mikity syndrome, complement levels other than C1q were significantly elevated in comparison with those of infants with respiratory distress syndrome. In the group of preterm infants without amnionitis, no differences were found between infants with intrauterine growth retardation and those with growth appropriate for gestational age.

Complement activation in fetuses: assessment by the levels of complement components and split products in cord blood.

The whole complement activity (CH50) and concentrations of components (C1q, C3, and C4) and C3 split product (C3d) were determined in the cord blood of 172 newborns between 16 and 42 weeks of gestation. Histological examination of their placentas revealed that the extent of complement activation was significantly higher in those with some degree of inflammation in the amnion than in those with normal amnion. The correlation between the complement levels and gestational age was higher in those with no amnionitis. In some cases the level of C3d at an early gestational age was as high as that in adults. The C3 activation system seems to be well developed even before week 25 of gestation. Among the complement components, C3d was the most sensitive indicator of placenta inflammation. As placenta inflammation may reflect intrauterine infection, the measurement of C3d levels may be of diagnostic value.

Determination of complement breakdown fragments C3d and its subfragments in health and disease.

The breakdown products of the third component of complement in approximately 400 samples were measured by rocket immunoelectrophoresis and two-dimensional electrophoresis using the method of Brandslund et al [3]. It was confirmed that the measurement of the C3d level provides useful information on increased C3 consumption irrespective of the synthetic rate. Furthermore, three subfragments with C3d but without C3c antigenicity were distinguished, which were designated as C3d1, C3d2, and C3d3. The subfragment C3d3 which migrated to the most anodal side was a predominant component in the plasma from patients with autoimmune diseases. Little C3d3 subfragment was detected in normal plasma and in normal sera incubated in vitro for 24 hr. Even in the normal sera converted completely in vitro which contained little intact C3, only a limited amount of C3d3 was detected. In the plasma from postsurgical patients in whom activation of the complement system was considered to be in an acute phase, C3d3 was detected, but the C3d2 level was higher than the C3d3 level. In the plasma from patients with systemic lupus erythematosus having the normal C3d level, C3d3 was a major fragment. It is predicted that the preponderant presence of C3d3 in plasma could be the result of chronic continuous complement activation by immune complexes.