Neutrophil defensins but not interleukin-6 in vaginal fluid after preterm premature rupture of membranes predict fetal/neonatal inflammation and infant neurological impairment.

OBJECTIVE: To determine whether neutrophil defensins (HNP1-3) and interleukin-6 (IL-6) in vaginal fluid after preterm premature rupture of membranes predict fetal inflammatory response syndrome (FIRS), neurological impairment or chorioamnionitis. DESIGN: Prospective study. SETTING: Tertiary referral university hospital. POPULATION: Forty-two patients with preterm premature rupture of membranes at <32 weeks. METHODS: Levels of HNP1-3 and IL-6 were measured in vaginal fluid obtained by swabs. Mann-Whitney U-test was used to compare HNP1-3 and IL-6 levels in groups with vs. without FIRS, infant death or neurological impairment, and chorioamnionitis (p<0.05 significant). Logistic regression was used to control for potential confounders. Diagnostic accuracies of HNP1-3 and IL-6 were determined by receiver operator characteristics analysis. MAIN OUTCOME MEASURES: Fetal inflammatory response syndrome was defined as neonatal inflammation within 72 hours postpartum. Neurological impairment was defined as motor and/or tone abnormalities at one year of corrected age. Chorioamnionitis was diagnosed histologically. RESULTS: Levels of HNP1-3, but not IL-6, were higher in 12 cases of FIRS (p=0.019 and p=0.256, respectively). Levels of HNP1-3, but not IL-6, were higher in 14 cases of infant death or neurological impairment (p=0.015 and p=0.100, respectively) and, when only survivors were analyzed, in nine cases of neurological impairment (p=0.030 and p=0.187, respectively). Levels of HNP1-3 and IL-6 were higher in 29 cases of chorioamnionitis (p=0.005 and p=0.003, respectively). The differences remained significant after adjustment for gestational age. Levels of HNP1-3 predicted FIRS, infant death or neurological impairment and chorioamnionitis with an area under the curve of 0.75, 0.79 and 0.78, respectively. CONCLUSIONS: Elevated vaginal fluid HNP1-3 and IL-6 levels are associated with histological chorioamnionitis. Elevated HNP1-3 can also identify FIRS and predict infant death or neurological impairment.

Highly improved metabolic stability and pharmacokinetics of indium-111-DOTA-gastrin conjugates for targeting of the gastrin receptor.

The development of metabolically stable radiolabeled gastrin analogues with suitable pharmacokinetics is a topic of recent research activity. These imaging vectors are of interest because the gastrin/CCK2 receptor is highly overexpressed in different tumors such as medullary thyroid cancer, neuroendocrine tumors, and SCLC. The drawback of current targeting agents is either their metabolic instability or their high kidney uptake. We present the synthesis and in vitro and in vivo evaluation of 11 (111)In-labeled DOTA-conjugated peptides that differ by their spacer between the peptide and the chelate. We introduced uncharged but hydrophilic spacers such as oligoethyleneglycol, serine, and glutamine. The affinity of all radiopeptides was high with IC(50) values between 0.5 and 4.8 nM. The improvement of human serum stability is 500-fold within this series of compounds. In addition the kidney uptake could be lowered distinctly and the tumor-to-kidney ratio improved almost 60-fold if compared with radiotracers having charged spacers such as glutamic acid.

Guidance on current good radiopharmacy practice (cGRPP) for the small-scale preparation of radiopharmaceuticals.

This guidance is meant as a guidance to Part B of the EANM "Guidelines on Good Radiopharmacy Practice (GRPP)" issued by the Radiopharmacy Committee of the EANM (see www.eanm.org ), covering the small-scale "in house" preparation of radiopharmaceuticals which are not kit procedures. The aim is to provide more detailed and practice-oriented guidance to those who are involved in the small-scale preparation of, for example, PET, therapeutic or other radiopharmaceuticals which are not intended for commercial purposes or distribution.

Neutrophil and monocyte CD64 indexes, lipopolysaccharide-binding protein, procalcitonin and C-reactive protein in sepsis of critically ill neonates and children.

OBJECTIVE: To compare the diagnostic accuracy of neutrophil and monocyte CD64 indexes (CD64in and CD64im) for sepsis in critically ill neonates and children with that of lipopolysaccharide-binding protein (LBP), procalcitonin (PCT) and C-reactive protein (CRP). DESIGN AND SETTING: Prospective, observational study in a level III multidisciplinary neonatal and pediatric intensive care unit (ICU). PATIENTS: Forty-six neonates and 36 children with systemic inflammatory response syndrome (SIRS) and suspected infection, classified into two groups: those with bacterial sepsis (microbiologically proven or clinical sepsis) and those without bacterial sepsis (infection not supported by subsequent clinical course, laboratory data and microbiological tests). INTERVENTIONS AND MEASUREMENTS: Flow cytometric CD64in and CD64im, serum LBP, PCT and CRP measurement on 2 consecutive days from admission to the ICU. RESULTS: There were 17 cases of bacterial sepsis in neonates and 24 cases of bacterial sepsis in children. All neonates and the majority of children were mechanically ventilated, and more than two-thirds of neonates with sepsis and one-third of children with sepsis needed inotropic/vasopressor drugs. The highest diagnostic accuracy for sepsis on the 1st day of suspected sepsis was achieved by LBP in neonates (0.86) and by CD64in in children (0.88) and 24 h later by CD64in in neonates (0.96) and children (0.98). CONCLUSIONS: Neutrophil CD64 index (CD64in) is the best individual marker for bacterial sepsis in children, while in neonates the highest diagnostic accuracy at the time of suspected sepsis was achieved by LBP and 24 h later by CD64in.

99mTc-labelled rituximab, a new non-Hodgkin's lymphoma imaging agent: first clinical experience.

OBJECTIVE: This study was performed to explore the possibility of using Tc-rituximab as an imaging agent to assess expression of CD20 antigen in patients with B-cell non-Hodgkin's lymphoma (NHL) before (radio) immunotherapy, for staging and subsequent evaluation of remission of NHL. METHODS: Rituximab was purified from Mabthera and photoactivated by ultraviolet light. The irradiated solution was aliquoted and labelled with pertechnetate. The effectiveness of the labelling method was evaluated by determination of the number of free thiol groups per photoreduced antibody, radiochemical purity determination and in-vitro stability. Immunoreactivity of Tc-rituximab was assessed on Ramos cells using a direct binding assay. Ten patients (age 31-70 years, mean 50 years) were included, nine with CD20 B-cell NHL and one with CD20-NHL. Whole-body and single photon emission computed tomography images were taken 1, 3, 6 and 20 h postinjection of Tc-rituximab. Scintigraphic results were compared with computerized tomography (CT) findings. RESULTS: In all cases radiochemical purity over 95% was observed with preserved affinity for CD20 antigen. In all patients expected activity was seen in the blood pool, liver, kidneys and spleen. Pathological, moderately to markedly increased Tc-rituximab activity was seen in all but one CT-confirmed NHL involved sites 6 and 20 h postinjection. In one patient, increased activity of Tc-rituximab was additionally seen in one region not seen on CT. In three patients increased accumulation was seen in bone marrow. CONCLUSION: Tc-rituximab is a promising imaging agent suitable for assessing expression of CD20 in patients with NHL before (radio) immunotherapy.

Colloidal 90Y used for radiation synovectomy decreases the viability of chondrocytes in human cadaveric samples.

OBJECTIVE: To analyse the early effect of colloidal 90Y beta irradiation, as used for radiation synovectomy (RSV), on the viability of human chondrocytes in an ex-vivo model. METHODS: Twenty osteochondral plugs (6 mm in diameter) were procured from femoral condyles of an adult male donor and stored in normal saline at 4 degrees C. The cartilage surfaces of 10 plugs were contaminated with colloidal 90Y citrate corresponding to the standard knee RSV dose (185 MBq) matched for the sample size (430 kBq). The remaining 10 plugs served as controls. At days 1, 2, 3, 6 and 13, two osteochondral plugs from each group were stained for viability with live/dead probes and scanned under a confocal laser microscope. The ratios of viable (green channel) and non-viable (red channel) pixels were acquired in four cartilage depth regions and statistically analysed with a regression model. RESULTS: The irradiation did not significantly alter the viable/non-viable pixel ratio during the first 2 days, but longer exposures led to a significant and time progressive reduction from 8.7% (day 3) to 12.5% (day 13). The ratio was less affected deeper in the cartilage, where it increased about 1% for every 100 microm from the surface. CONCLUSIONS: Surface exposure of human cadaveric cartilage to a therapeutic dose of colloidal 90Y decreased chondrocyte viability, expressed as the viable/non-viable pixel ratio, in the early post-irradiation period. The findings established in the ex-vivo simulation may reflect the changes in knee cartilage occurring after RSV therapy.

Activity or mass concentration of bone-specific alkaline phosphatase as a marker of bone formation.

BACKGROUND: Recently published data identified bone-specific alkaline phosphatase (BALP) as a good marker of bone formation in different bone diseases and osteoporosis. Two methods are available for BALP determination: one measures enzyme activity, the other its mass concentration. We compared results for BALP activity and its mass concentration in a group of 88 healthy pre- and postmenopausal women to identify which is a more useful marker for detecting early menopausal bone remodelling changes. METHODS: We measured BALP activity and BALP mass concentration in relation to femoral neck (FN) and lumbar spine (LS) bone mineral density (BMD) and some other widely used bone markers: osteocalcin (OC), procollagen type I N-terminal propeptide (PINP) and serum C-terminal telopeptide cross-links of type I collagen (CTx) in serum samples from 50 premenopausal (age 45.9+/-4.6 years) and 38 postmenopausal (age 54.4+/-4.5 years) women. RESULTS: Healthy postmenopausal women exhibited 34.2% (p<0.01) and 27.3% (p=0.000) higher levels of BALP activity and mass concentration than premenopausal women, respectively. At the same time, FN and LS BMD were not significantly different between the groups. CTx values were significantly higher in postmenopausal women (p=0.018), while OC and PINP were not. We observed significant correlation between BALP activity and mass concentration (r=0.85, p<0.01). The correlation between BALP activity and FN BMD or LS BMD was insignificant. BALP mass correlated significantly with LS BMD (r=-0.370, p=0.033) but not with FN BMD. As expected, we proved a significant positive correlation for both BALP methods with the other bone markers measured in our study. CONCLUSIONS: Postmenopausal women have slightly higher bone turnover. Since LS and FN BMD were not significantly lower in our group of healthy postmenopausal women, but BALP and CTx were markedly higher, we suggest that measurements of BALP and CTx might be useful as early markers of higher bone turnover. Finally, our results did not show any differences between the clinical utility of BALP activity and BALP mass concentration measurements.