Genetic and phenotypic relationships between immune defense, melanism and life-history traits at different temperatures and sexes in Tenebrio molitor.

Insect cuticle melanism is linked to a number of life-history traits, and a positive relationship is hypothesized between melanism and the strength of immune defense. In this study, the phenotypic and genetic relationships between cuticular melanization, innate immune defense, individual development time and body size were studied in the mealworm beetle (Tenebrio molitor) using three different temperatures with a half-sib breeding design. Both innate immune defense and cuticle darkness were higher in females than males, and a positive correlation between the traits was found at the lowest temperature. The effect of temperature on all the measured traits was strong, with encapsulation ability and development time decreasing and cuticle darkness increasing with a rise in temperature, and body size showing a curved response. The analysis showed a highly integrated system sensitive to environmental change involving physiological, morphological and life-history traits.

Assessment of the tibia using ultrasonic guided waves in pubertal girls.

The purpose of this study was to compare low frequency ultrasonic guided wave measurements with established ultrasound and bone density measurements in terms of their ability to characterize the tibia in pubertal girls. Subjects were 12-14-year-old girls ( n=106) who were participating in a calcium and vitamin D intervention study. A prototype low frequency pulse transmission device consisting of a uniaxial scanning mechanism and low frequency transducers orientated perpendicularly to the limb was used to measure two ultrasound velocities in the tibia. The first velocity, V1, was that of the first arriving signal, similar to that measured by existing commercial tibial ultrasound devices. The second velocity, V2, was that of a slower wave propagating at 1,500-2,000 m/s, which has been shown elsewhere to be consistent with the lowest order antisymmetric guided mode in the bone. In addition, commercial ultrasound devices (Omnisense, Sunlight Ltd.; QUS-2, Quidel Corp.) were used to measure the speed of sound (SOS) in the tibia and the radius and attenuation (BUA) in the calcaneus. Cortical bone cross-sectional area (CSA), mineral density (BMD) and cortical thickness (cTh) of the tibia were measured using pQCT, site-matched to the ultrasound measurements. Both V1 and V2 correlated significantly with cortical BMD and with cTh and CSA. On the other hand, tibial SOS correlated with BMD, but not with cTh and CSA. These results indicate that the prototype device using guided waves captures aspects of tibial cortical bone geometry in addition to bone density, thereby potentially offering increased diagnostic information compared to existing tibial ultrasound devices.

Comparison of media for enumeration of Clostridium perfringens from foods.

Many media have been developed to enumerate Clostridium perfringens from foods. In this study, six media [iron sulfite (IS) agar, tryptose sulfite cycloserine (TSC) agar, Shahidi Ferguson perfringens (SFP) agar, sulfite cycloserine azide (SCA), differential clostridial agar (DCA), and oleandomycin polymyxin sulfadiazine perfringens (OPSP) agar] were compared in a prestudy, of which four (IS, TSC, SCA, and DCA) were selected for an international collaborative trial. Recovery of 15 pure strains was tested in the prestudy and recovery of one strain from foodstuffs was tested in the collaborative trial. Results from the prestudy did reveal statistical difference of the media but recoveries on all media were within the microbiological limits (+/-30%) of IS, which was set as a reference medium. Recoveries on the media tested in the collaborative trial were statistically different as well, but these differences were of no microbiological-analytical relevance. Food matrices did not affect the recovery of C. perfringens in general. DCA and SCA, in particular, are labor-intensive to prepare and DCA frequently failed to produce black colonies; gray colonies were quite common. Since IS medium is nonselective, it was concluded that TSC was the most favorable medium for the enumeration of C. perfringens from foods.

Combination of cervical interleukin-6 and -8, phosphorylated insulin-like growth factor-binding protein-1 and transvaginal cervical ultrasonography in assessment of the risk of preterm birth.

OBJECTIVE: To determine the value of combinations of cervical interleukin-6 (IL-6), cervical interleukin-8 (IL-8), the phosphorylated isoform of insulin-like growth-factor binding protein-1 (IGFBP-1), and cervical ultrasonography in the prediction of preterm birth. DESIGN: Prospective follow up. SETTING: Oulu University Hospital maternity clinic from February 1997 to July 1998. POPULATION: Women with singleton pregnancies (n = 77), referred from outpatient clinics at 22-32 weeks of gestation with symptoms (uterine contractions) or signs (cervical change) of threatened preterm birth. Symptomless women (n = 78) matched for gestational age, parity and maternal age at recruitment were studied as a reference group. METHODS: A urine sample for bacterial culture was collected, and cervical swab samples for assays of interleukin-6 and -8 and phoshorylated IGFBP-1 were taken before digital cervical examination. A Pap smear for analysis of bacterial vaginosis and samples for analysis of chlamydia and streptococci were also obtained. Cervical measurements were made by transvaginal ultrasonography. The same sampling and cervical measurement were repeated twice at two-week intervals. The cutoff values of the markers were determined by receiver-operating characteristic curve analysis. MAIN OUTCOME MEASURE: Preterm birth (<37 weeks). RESULTS: The preterm birth (<37 weeks) rate for women in the study group was 16% (12/77). The cervical interleukin-6 cutoff value (61 ng/L) at first visit had a sensitivity of 73% and a specificity of 61% in predicting preterm birth, with a positive likelihood ratio (LR+ ) of 1.9 (95% CI 1.2-3.0). An ultrasonographically measured cervical index value of > 0.36 at recruitment predicted preterm birth in 25% (5/20) of the study group compared with 9% (5/54); LR+ 2.2 (95% CI 1.03-4.7). Cervical phosphorylated IGFBP-1 > 6.4 microg/L [LR+ 1.8 (95% CI 0.7-2.9)], interleukin-8 > 3739 ng/L [LR+ 1.4 (95% CI 0.9-2.4)], and ultrasonograpic cervical length < 29.3 mm [LR+ 2.7 (95% CI 0.8-9.7)] increased the risk of preterm birth. According to the logistic regression model, a combination of IL-6, and IL-8 and cervical index increased the specificity to 97%, but the sensitivity fell to 30% in detecting preterm birth. There was a significantly increased incidence of puerperal infections if phosphorylated IGFBP-1 concentrations were elevated (> 21.0 microg/L), 36% (4/11) compared with 4.6% (3/65), LR+ 6.7 (95% CI 2.7-17), the sensitivity being 67% (4/6) and the specificity 90% (63/70). Elevated phosphorylated IGFBP-1 concentrations (> 21.6 microg/L) were also associated with an increased risk of neonatal infections; LR+ 8.0 (95% CI 3.5-18). CONCLUSIONS: An increase in cervical IL-6 concentration and the ultrasonographically measured cervical index appear to be associated with preterm birth. A combination of these markers with measurement of cervical IL-8 appears to be the best predictor of preterm birth. Neither the sensitivity nor specificity of the tests used in this study are good enough to predict preterm birth for clinical decision making. Cervical phosphorylated IGFBP-1 seems to be a marker of puerperal and neonatal infectious morbidity in cases of threatened preterm delivery, suggesting early tissue degradation at the choriodecidual interface.

Insulin-like growth factor-binding protein-1 in cervical secretion as a predictor of preterm delivery.

BACKGROUND: The aim of the study was to evaluate whether the phosphorylated isoforms of insulin-like growth factor-binding protein-1 (IGFBP-1), a protein produced by the decidua, can be detected in cervical secretions of pregnant women with preterm uterine contractions, and whether their presence predicts an increased risk of preterm delivery. METHODS: A prospective analysis of sixty-three women who presented with preterm labor but intact fetal membranes at weeks 22-36+6 days of gestation at the Antenatal clinic at the Department of Obstetrics and Gynecology, Helsinki University Central Hospital. Phosphorylated IGFBP-1 (phIGFBP-1) was measured in cervical swab samples obtained at presentation, using an immunoenzymometric assay. The values > or =10 microg/L were considered as positive. In addition, 58 asymptomatic women at the same gestational stage were studied as controls. Multiple logistic regression was applied to control for confounding variables and to obtain adjusted odds ratios. RESULTS: The concentration of phIGFBP-1 in cervical samples ranged from undetectable to 95 microg/L. In 17 of the 63 (27%) women with preterm labor it was > or =10 microg/L. Seven of these 17 (41%) women with a positive phIGFBP-1 result delivered preterm, all before 35 weeks of gestation. Among the women with preterm labor and a negative phIGFBP-1 result, three of the 46 (7%) delivered before 37 weeks of gestation (adjusted OR 24, 95% CI 1.2-487), but all after 35 weeks of gestation. In the asymptomatic control population three out of 58 (5%) women had a positive cervical phIGFBP-1 test result but none delivered preterm. Among the controls with a negative cervical phIGFBP-1 test result (55 of 58, 95%), one woman delivered preterm (1 of 55, 2%). CONCLUSIONS: Pregnant women who are in preterm labor with intact fetal membranes and who have a positive phIGFBP-1 test result in cervical secretion have an increased risk of preterm delivery.

The effect of refrigeration on the osmolality and pH of nonpreserved artificial tears containing carboxymethylcellulose.

INTRODUCTION: Nonpreserved artificial tears (NPAT) are a recommended treatment for dry eye. The manufacturers' instructions state to discard the container after initial opening and use. Some clinicians advocate the use and storage of NPAT in a zip-lock bag in a refrigerator for up to 12 h. The purpose of this study was to evaluate whether refrigeration of opened NPAT over a 12-h period had any effect on the pH or osmolality. METHODS: Forty individual carboxymethylcellulose NPAT samples were used in this study. The initial osmolality and pH of each sample were measured with a vapor pressure osmometer and electronic pH meter. The samples were refrigerated (4 degrees C) in closed zip-lock plastic bags for 12 h. After storage, the pH and osmolality of the samples were measured. The data were statistically analyzed for significant differences using a paired t-test. RESULTS: The mean initial pH and osmolality before refrigeration were 6.46 pH units and 304.10 mmol/kg, respectively. After refrigeration, the mean pH was 6.44 units, and mean osmolality was 305.87 mmol/kg. Paired t-tests revealed a nonsignificant difference (p > 0.05) for both pH and osmolality. CONCLUSION: Refrigeration of opened carboxymethylcellulose NPAT stored in closed zip-lock plastic bags does not have a significant effect on the osmolality or pH of the solution. Storage of NPAT containing carboxymethylcellulose is an acceptable practice with regards to stability of pH and osmolality.

Phosphorylated isoforms of insulin-like growth factor binding protein-1 in the cervix as a predictor of cervical ripeness.

OBJECTIVE: To study the isoforms of insulin-like growth factor binding protein-1 (IGFBP-1) in cervical secretion and to evaluate whether their assessment could serve in prediction of cervical ripeness at term. METHODS: We measured the concentrations of IGFBP-1 in cervical swab samples of 64 women scheduled for labor induction by amniotomy or cervical ripening with prostaglandin E2 gel. Two immunoenzymometric assays were used: a previously described assay 1, which detects the nonphosphorylated and lesser phosphorylated isoforms, and a novel assay 2, which detects the lesser and highly phosphorylated isoforms of IGFBP-1. A set of 39 amniotic fluid (AF) samples also was analyzed to compare the phosphorylation status of IGFBP-1 in cervical secretion with that in AF. RESULTS: In all cervical samples, IGFBP-1 concentration was higher by assay 2 than by assay 1, whereas in all AF samples, the results were the opposite. Initially, the median IGFBP-1 concentration in the ripe cervices (Bishop scores 6 or greater; n = 29) was approximately four times as high as that in the unripe cervices (Bishop scores 5 or less; n = 35). The cervical IGFBP-1 concentrations increased eight-fold in 6 hours after the first application of PGE2. CONCLUSION: Phosphorylated isoforms of IGFBP-1, different from those in AF, are present in the cervical secretion of women with intact fetal membranes and reflect cervical ripeness. A bedside test for those IGFBP-1 isoforms might help in predicting amenability for labor induction.

Ocular tolerability of timolol in Gelrite in young glaucoma patients.

BACKGROUND: Although initial patient response to timolol maleate in Gelrite (Timoptic XE) has been generally favorable in clinical settings, anecdotal reports of blurred vision warrant further investigation comparing the tolerability of this new formulation with that of timolol maleate in conventional solution. METHODS: A prospective, double-masked, randomized, cross-over study was performed using 28 normal volunteers who had no evidence of ocular disease, including glaucoma. Mean subject age was 32.5 years. Subjects were randomized to receive one drop in each eye of either timolol maleate in solution (TS) 0.5 percent or timolol maleate in Gelrite (TXE) 0.5 percent. Subjects rated each medication, using a visual analog scale, at the following time intervals: prior to drug instillation, immediately following instillation, 1 min, and at 15 min, 30 min, 1 hour, 2 hours, and 4 hours. RESULTS: Wilcoxon Matched Paris Test showed statistically significant differences at 1 minute for tearing (p = 0.003), blurred vision (p = 0.001) and drug acceptance (p = 0.028) in favor of TS. At 15 minutes and thereafter, however, none of the differences between groups was significant. Visual acuities were unaffected by either TS or TXE. CONCLUSIONS: These data suggest that TXE may impair visual function more than TS, but these effects are briefly and transitory. TXE appears to be a clinically acceptable drug delivery vehicle for use during the young glaucoma patient's waking hours.

Evaluation of a rapid strip test for insulin-like growth factor binding protein-1 in the diagnosis of ruptured fetal membranes.

We evaluated the clinical usefulness of a new bedside test (PROM TEST) for insulin-like growth factor binding protein-1 (IGFBP-1) in the detection of ruptured fetal membranes (ROM). Cervicovaginal secretion was sampled between 15 and 37 weeks of gestation from asymptomatic women with apparently intact membranes and from women with clinically confirmed ROM, as well as from symptomatic women with suspected ROM based on history. IGFBP-1 in samples was detected with a dipstick based on immunochromatography. The test result was positive in 100% of cases with unequivocal ROM and in 5.3% of cases with apparently intact membranes. Furthermore, the PROM TEST was positive in 64 of 181 patients evaluated for suspected ROM based on history, but in whom the diagnosis could not be clinically confirmed at the initial evaluation. Fifty of the 64 women (78.1%) were delivered prematurely (< 37 weeks). Five of the 117 PROM-negative patients had elective cesarean section for reasons unrelated to ROM before 37 weeks and 10 of the remaining 112 patients (8.9%) had preterm delivery. Women with equivocal ROM and a positive test result had a 6.9-fold increased relative risk (95% confidence interval 4.2-11.4) of preterm delivery compared with women who had a negative result at the time of evaluation. Multiple logistic regression including PROM TEST result, contractions, vaginal bleeding and cervical changes indicated that a positive PROM TEST result was an independent predictor of preterm delivery (P = 0.0001). In summary, a positive PROM TEST result identifies ROM with high sensitivity and a negative result effectively excludes those with intact membranes. In patients with suspected but clinically unconfirmed ROM, the positive test result is associated with increased risk of preterm delivery, suggesting that microruptures of fetal membranes can also be detected by the PROM TEST.

Aging is associated with decreased suppression of insulin-like growth factor binding protein-1 by insulin.

We determined whether aging influences circulating insulin-like growth factor-binding protein-1 (IGFBP-1) concentrations and, if so, whether this effect is explained by altered regulation of IGFBP-1 by insulin. Fasting levels of glucose, insulin, and IGFBP-1 were measured in 94 healthy volunteers, ages 24-93 yr. To determine the effect of aging on insulin-induced suppression of IGFBP-1, an oral glucose tolerance test (OGTT) was performed in 10 older (72-92 yr) and 10 younger (24-58 yr) nonobese subjects, matched for sex and body mass index. For all ages combined, the mean glucose level (+/- SE) averaged 4.9 +/- 0.1 mmol/L, and there was no significant change with aging. Fasting insulin and IGFBP-1 concentrations increased with advancing age (r = 0.37, P < 0.001 for age vs. insulin and r = 0.47, P < 0.001 for age vs. IGFBP-1). However, there was no correlation between insulin and IGFBP-1 concentrations. In multiple linear regression analysis, the age-related increase in IGFBP-1 was independent of body mass index. During the OGTT, the mean insulin concentration was significantly higher in the older group compared with the younger group (P < 0.001). Serum IGFBP-1 concentrations were higher in the fasting state as well as during the OGTT, and the mean percent decrease of IGFBP-1 below baseline was significantly smaller in the older compared to the younger subjects at 3 h (35 +/- 5% vs. 55 +/- 2%, P < 0.01). We conclude that aging is associated with decreased suppression of serum IGFBP-1 by insulin, as demonstrated by 1) lack of the inverse correlation between fasting insulin and IGFBP-1 seen in young adults; 2) concurrent elevation of fasting insulin and IGFBP-1 concentrations; and 3) a blunted decrease in serum IGFBP-1 during an OGTT.

Relationship between carbohydrate metabolism and serum insulin-like growth factor system in postmenopausal women: comparison of endometrial cancer patients with healthy controls.

Insulin is a major regulator of circulating insulin-like growth factor (IGF)-binding protein-1 (IGFBP-1), suppressing the hepatic production of IGFBP-1. Postmenopausal age, obesity, hypertension, and impaired glucose tolerance, which are known risk factors for endometrial cancer, are all associated with hyperinsulinemia and insulin resistance. In this study, we investigated the relationship among serum insulin, glucose, insulin-like growth factors (IGF-I and IGF-II), and IGFBP-, -2, and -3 in 32 nondiabetic postmenopausal women with endometrial cancer and in 18 healthy controls. The mean fasting levels of glucose and insulin were higher, whereas the mean basal IGF-I, IGF-II, and IGFBP-3 levels were lower in the endometrial cancer patients than in the healthy control subjects. The mean fasting IGFBP-1 and IGFBP-2 levels did not differ between the groups, and no correlation was found between fasting insulin and IGFBP-1 concentrations or between insulin and IGFBP-2 concentrations in either of the study groups. During an oral glucose tolerance test, the mean glucose levels at 1 and 3 h as well as the mean insulin level at 3 h were significantly higher in the endometrial cancer patients than in the controls, and the area under the glucose curve was larger in the first group. An oral glucose load resulted in a similar fall in serum IGFBP-1 levels in endometrial cancer patients and controls (51% and 55% at 3 h). When the cancer patients were divided into two subgroups according to the body mass index (kilograms per m2), the obese group had higher glucose and insulin indices than the nonobese group. No difference was found by the same measures in healthy controls. The fasting serum IGFBP-1 levels tended to be lower in the obese than in the normal weight subjects, but the difference did not reach statistical significance. In summary, these results provide preliminary evidence that the inverse relation between fasting insulin and IGFBP-1, well established in children and young adults, disappears in elderly women, although short term suppression by insulin still occurs. Further, our data indicate that in addition to carbohydrate metabolism, postmenopausal women with endometrial cancer have alterations in their circulating IGF system compared to controls.

Measurement of insulin-like growth factor binding protein-1 in cervical/vaginal secretions: comparison with the ROM-check Membrane Immunoassay in the diagnosis of ruptured fetal membranes.

Insulin-like growth factor binding protein-1 (IGFBP-1) is a major protein in amniotic fluid. In this study, we evaluated the diagnostic potential of IGFBP-1 measurement in cervical/vaginal secretions as an indicator of ruptured fetal membranes. Data were also compared with those obtained by the ROM-check Membrane Immunoassay (Adeza Biochemical, Sunnyvale, California) that is based on the detection of fetal fibronectin in vaginal fluid. In women with intact membranes and not in labor, the concentration of IGFBP-1 in specimens obtained from the cervix and immersed in 0.5 ml of assay buffer ranged from < 0.5 to 90 micrograms/l, whereas in specimens obtained less than 8 h after spontaneous or artificial rupture of membranes it varied between 175 and 20,000 micrograms/l, the median being 1,900 micrograms/l. The values greater than 100 micrograms/l were interpreted as containing amniotic fluid. The IGFBP-1 measurement and the ROM-check Membrane Immunoassay were carried out parallel in the vaginal swab specimens obtained from 54 pregnant women from 1 h to 1 week after the rupture. Twenty-four women had ROM confirmed from 1 h to 1 week earlier. In this group of patients, IGFBP-1 concentration > 100 micrograms/l had a sensitivity of 75% and a specificity of 97% in diagnosis of ROM. The corresponding numbers for a positive ROM-check were 92% and 80%. The positive predictive value was 95% for the IGFBP-1 measurement compared to 79% for the ROM-check test.(ABSTRACT TRUNCATED AT 250 WORDS)

Rapid determination of C-reactive protein by enzyme immunoassay using two monoclonal antibodies.

Several monoclonal antibodies for human C-reactive protein (CRP) were characterized, and two antibodies binding to separate domains were used to construct a rapid and simple immunoenzymometric assay for CRP. The assay consists of a single 15 min immunological reaction during which CRP forms a complex with a peroxidase-labelled antibody and with another antibody attached to the test-tube wall. The immobilized complex is detected by a 3 min colour reaction using peroxidase substrate. The quantitative measuring range of the assay is 0.04-5 mg/l, and no hook occurs at five-fold higher values. The sensitivity of the method allows reliable determination of low CRP levels, eg. in paediatric samples. The values obtained with the present assay correlated well with turbidimetric results.

A monoclonal antibody-based immunoradiometric assay for low molecular weight insulin-like growth factor binding protein/placental protein 12.

We describe a sensitive immunoradiometric assay for insulin-like growth factor binding protein/Placental Protein12 (IGF-BP/PP12) using monoclonal antibodies. This assay has a detection limit of 0.25 micrograms/l IGF-BP/PP12. Parallel dose response curves were obtained with purified IGF-BP/PP12, amniotic fluid, decidual cytosol extract, and serum. The assay is reproducible (intra-assay variation 4.3-8.2% and interassay variation 9.7-11.1%) and fast (less than 5 hours). A crossreactivity of less than 0.01% for all other proteins tested reflects high specificity. Using this method the mean serum IGF-BP/PP12 concentration in healthy women was 5.2 micrograms/l. During pregnancy, the mean IGF-BP/PP12 at 7-11 weeks was 43.3 micrograms/l, and at 36-40 weeks 121 micrograms/l. After early pregnancy termination the serum IGF-BP/PP12 decreased rapidly reaching a mean level of 8 micrograms/l within 4 days.

Monoclonal antibodies to the 27-34K insulin-like growth factor binding protein.

Monoclonal antibodies were prepared against the 27-34K insulin-like growth factor (IGF)-binding protein purified from human placenta/decidua and designated placental protein 12 (PP12). Four different antibodies were characterized. Each recognized the major band at 32K on immunoblots of the purified PP12 preparation and amniotic fluid. In liquid phase RIA, IGF-I did not affect the binding of [125I] PP12 to one antibody (Mab 6303), it slightly increased the binding to two antibodies (Mab 6301 and 6304), and it slightly decreased the binding to one antibody (Mab 6302). All antibodies immunoprecipitated the cross-linked PP12-[125I] IGF-I complex, but Mab 6302 considerably less effectively than the others. Preincubation of PP12 with Mab 6302 completely inhibited the binding of [125I] IGF-I to PP12, whereas preincubation with Mab 6303 had no effect, and Mab 6301 as well as Mab 6304 increased it. These results suggest that Mab 6302 binds to an epitope at or near to the IGF-binding site, whereas the other antibodies react at other sites of the PP12 molecule. Conformational changes in PP12 probably account for the IGF-I-induced increase in the binding of Mabs 6301 and 6304 to [125I] PP12, and vice versa, for Mabs 6301- and 6304-induced increase in the binding of [125I] IGF-I to PP12.