Diagnostic value of immature myeloid information in early-onset bacterial infection in term and preterm neonates.

BACKGROUND: For quick detection of neonatal early-onset bacterial infection (EOBI) pro-inflammatory cytokines like Interleukin-6 (IL-6) and Interleukin-8 (IL-8) in combiantion with C-reactive Protein (CRP) have been used. Automated determination of immature myeloid information (IMI) seems to be an additional useful tool in the diagnosis of NBI. OBJECTIVE: To compare the diagnostic value of IMI, I/T-Ratio, plasma IL-6 and IL-8 levels and CRP in term and preterm neonates at time of clinical suspicion of EOBI. PATIENTS AND METHODS: 31 preterm and 123 term neonates with clinical and serological signs of EOBI were analysed. 91 preterm and 159 term neonates with risk factors but without proven EOBI served as non-infected controls. RESULTS: Neonates with EOBI showed significantly elevated IMI levels at time of first clinical suspicion of EOBI (Preterm: 1 028/µL (38-8 759) vs. 289/µL (6-3 126); Term: 1 268/µL (48-14 035) vs. 856/µL (19-5 735); p<0.05 respectively). I/T-Ratio, IL-6, IL-8 and CRP values were significantly higher in preterm and term neonates with EOBI (p<0.05). Sensitivity of IMI at a cut-off level of 650/µL was 84.2% [95%-CI: 74.0-91.6%] in preterm and 65.4% [95%-CI: 56.8-73.3%] in term infants. Specificity was 66.7% [95%-CI: 47.1-82.7%] and 53.9% [95%-CI: 43.8-63.7%], respectively. Combination of different infection parameters improved sensitivity up to 93.5% and specificity up to 98.9%. CONCLUSION: The diagnostic value of IMI in diagnosing EOBI in preterm and term neonates is not comparable to IL-6, IL-8 and CRP. Combination of IMI-Channel with IL-6, IL-8 or CRP improves their sensitivity, specificity and predictive value.

Treatment of neonatal abstinence syndrome in preterm and term infants.

OBJECTIVE: Neonatal abstinence syndrome (NAS) is treated with a variety of drug preparations. With the optional treatment of NAS with chloral hydrate, phenobarbital or morphine the cumulative drug consumption of the mentioned drugs, the length of hospital stay and treatment duration was evaluated in preterm and term neonates. METHODS: Retrospective, uncontrolled study which evaluates different therapies of neonatal abstinence syndrome (NAS) in preterm and term neonates. RESULTS: During the past 16 years data were obtained from medical records of 51 neonates with NAS; 9 preterm and 35 term neonates were evaluated and 7 were excluded because of incomplete data sets. 31 (72.1%) received a pharmacological treatment (6 preterm and 25 term neonates). Treatment started at 4.3 [3.3-5.3] d. Mean duration of treatment was 11.7 [6.6-16.7] d. In our study chloral hydrate (ch) and phenobarbital (pb) were first line medication escalated by the morphine (mp) solution. Mean cumulative dosage of ch was 643.5 [260.3-1 026.7] mg, of pb 53.2 [19.7-86.8] mg and of mp 4.22 [0-8.99] mg. CONCLUSION: Our study group showed similar treatment duration and length of hospital stay compared to other studies. The cumulative dose of mp was lower compared to most studies. This benefit resulted at the expense of a further medication with pb and ch. However, 6 of 9 preterm neonates needed significantly less pharmacological therapy compared to term neonates indicating less susceptibility of immature brain to abstinence of maternalo-pioids.

Proteinase-activated receptor-4 evoked colorectal analgesia in mice: an endogenously activated feed-back loop in visceral inflammatory pain.

BACKGROUND: Activation of proteinase-activated receptor-4 (PAR-4) from the colonic lumen has an antinociceptive effect to colorectal distension (CRD) in mice in basal conditions. We aimed to determine the functional localization of the responsible receptors and to test their role in two different hyperalgesia models. METHODS: Mice received PAR-4 activating peptide (PAR-4-AP, AYPGKF-NH(2)) or vehicle intraperitoneally (IP), and abdominal EMG response to CRD was measured. The next group received PAR-4-AP intracolonically (IC) with or without 2,4,6-triaminopyrimidine, a chemical tight junction blocker, before CRD. The SCID mice were used to test the role of lymphocytes in the antihyperalgesic effect. The effects of PAR-4-AP and PAR-4-antagonist (P4pal-10) were evaluated in water avoidance stress (WAS) model and low grade 2,4,6-trinitrobenzene sulfonic acid (TNBS) colitis. Spinal Fos protein expression was visualized by immunohistochemistry. KEY RESULTS: The antinociceptive effect of PAR-4-AP disappeared when was administrered IP, or with the blockade of colonic epithelial tight junctions, suggesting that PAR-4-AP needs to reach directly the nerve terminals in the colon. The CRD-induced spinal Fos overexpression was reduced by 43% by PAR-4-AP. The PAR-4-AP was antihyperalgesic in both hyperalgesia models and in mice with impaired lymphocytes. The PAR-4-antagonist significantly increased the TNBS, but not the WAS-induced colonic hyperalgesia. CONCLUSIONS & INFERENCES: The antinociceptive effect of PAR-4-AP depends on its penetration to the colonic mucosa. The PAR-4 activation is endogenously involved as a feedback loop to attenuate inflammatory colonic hyperalgesia to CRD.

Stable precursor fragments of vasoactive peptides in umbilical cord blood of term and preterm infants.

BACKGROUND: Though various neurohormonal systems are concurrently activated during birth, their biological effectors are not always easy to measure due to their short half-life in vivo, instability in biological samples, or very low concentrations. METHODS: Using a recently discovered chemiluminescence assay, we measured the stable precursor fragments mid-regional pro-atrial natriuretic peptide (MR-proANP), mid-regional pro-adrenomedullin (MR-proADM), C-terminal pro-endothelin-1 (CT-proET-1) and C-terminal pro-vasopressin (CT-proAVP or copeptin) immediately after birth in umbilical venous cord blood from 119 infants with a gestational age of 23-42 weeks and evaluated their possible functions. RESULTS: Cord blood levels of MR-proANP, MR- proADM, CT-proET-1, and CT-proAVP were considerably higher compared with normal adult levels. The CT-proAVP concentrations were 10-fold higher in term, and 70-fold higher in extremely preterm infants. MR-proANP showed 4-fold higher levels in term infants and 20-fold higher levels in extremely preterm infants. Levels of MR-proADM and CT-proET-1 were 2- to 3-fold higher in preterm and term infants. All four parameters showed significantly decreased values with increasing gestational age and a significant correlation between CT-proET-1 and MR-proADM. CONCLUSION: Our results indicate that vasoactive and natriuretic mediators play a functionally relevant role in circulatory transition from fetal to neonatal life.

alpha-Ketoglutarate (AKG) absorption from pig intestine and plasma pharmacokinetics.

To study the absorption, metabolism and kinetics, the AKG (in different concentrations) was administered intravenously, intra-portally, orally and directly into the ileum or duodenum of pigs, chronically fitted with portal and jugular catheters and T-shaped cannula at the duodenum and ileum. Additionally, this study was conducted to determine the influence of low pH, Fe(2+) or/and SO on AKG gut absorption and conversely FeSO(4) and FeSO(4)/AKG on Fe(2+) gut absorption. It is concluded that AKG was significantly better absorbed from the upper small intestine than from the distal sections. Furthermore, low pH, Fe(2+) and/or SO ions enhanced AKG absorption. The AKG administered to the portal vein was rapidly eliminated from the blood (half-life less than 5 min). The short lifetime for AKG is probably dependent on quick metabolism in the enteorcyetes and liver. However, the prolonged half-life can be related to its low AKG blood concentration. The Fe(2+) concentrations in blood increased after FeSO(4) and FeSO(4)/AKG duodenal infusion. The implication of above observations is important for practical application of the AKG in animal and human nutrition as well in medicine.

Occupational exposure to external ionising radiation in Poland, 1999.

In 1999 about 6208 radiation workers from 389 departments were monitored by CLOR in Poland. The distribution of annual personal doses shows that 85% of controlled workers received doses below the MDL (0.4 mSv) and about 97% controlled workers received doses below 5 mSv. Doses higher than 50 mSv were received by three operators of industrial radiography units. The radiation workers under control are divided into four main work sectors: nuclear industry, research and education, medicine, and general industry. The average annual dose for all workers in each particular sector was 0.22 mSv, 0.22 mSv, 0.30 mSv and 0.80 mSv, respectively. The average annual dose for the entire monitored population was 0.47 mSv. The average annual dose in each particular sector for number of workers receiving E > 0, i.e. Hp(10) > or = 0.4 mSv, amounted to 1.78 mSv, 2.03 mSv, 1.88 mSv and 4.85 mSv, respectively. The average annual dose for the full number of workers receiving E > 0 was 3.21 mSv. This paper contains the detailed analysis of occupational exposure. The distributions of annual occupational exposure in different work sectors are also given.

A high resolution PIXE measurement for blood plasma ultrafiltrate. Application to loosely bound copper.

A new technique for investigation of elemental concentrations in subfractions of blood plasma is presented. The method is composed of the ultrafiltration of plasma in the presence of ethylenediaminetetraacetic acid (EDTA) and the measurement of the elemental composition by proton-induced X-ray emission (PIXE). The blood samples were collected from both healthy persons and patients suffering from breast cancer. The main emphasis in this study was on the determination of loosely bound copper (Cu) in plasma subfractions containing substances with molecular mass under 10,000, but zinc (Zn) and iron (Fe) contents of these fractions were also determined. The detection limits obtained with this method for Cu, An and Fe were approximately 10 ppb (wet wt).

Early breast cancer, diet, and plasma copper fractions.

In a study of diet and early breast cancer, blood plasma copper has been analyzed by Proton Induced X-ray Emission analysis as both total copper (P-Cu) and that ultrafiltrable from plasma (P-edu-Cu) through membranes with a cut-off at molecular mass 10,000 after equilibration with disodium ethylene diamine tetraacetic acid (EDTA) at 4 degrees C. Ceruloplasmin (P-cer) was also measured using nephelometry of anticeruloplasmin monoclonal antibody-ceruloplasmin complexes. Dietary copper intake per day (D-Cu) was assessed over a five-day dietary record period and calculated from dietary components using a computer program. P-edu-Cu correlated significantly with both D-Cu and ceruloplasmin while P-Cu correlated only with ceruloplasmin. Further, ceruloplasmin did not significantly correlate to D-Cu. Hence, P-edu-Cu better reflects copper status than do P-cer or P-Cu as it relates to both the major copper enzyme in plasma and to daily copper intake. This may be important in drawing conclusions about the significance of copper in disease states where copper fractions other than ceruloplasmin may be most important owing, for example, to oxidative properties. Categorization as cancer or normal, by copper parameters (D-Cu, P-edu-Cu, P-Cu, P-cer), was studied in multiple correlation. In particular, the ratio P-cer/P-Cu and the ratio P-edu-Cu/D-Cu were significantly related to disease. Irrespective of age (pre- and post-menopausal), highly significant differences between normals and early stage breast cancer patients were seen with p < 0.0001 to p < 0.01. The precise role played by plasma and dietary copper fractions deserves continued attention in view of the present and earlier results in cancer studies.

Effects of preparation methods of biomedical samples on PIXE measurement sensitivity.

Instrumental sample preparation techniques for PIXE measurements of human blood serum and bovine muscle were investigated. The techniques studied were direct measurement of a drop, drying on a foil and on an Al cup, lyophilization on a foil, pellet pressing of dried material, microtome sectioning and biopsy preparation. The same serum and muscle were used in each procedure allowing the comparison of detection limits between different methods. The most beneficial technique for both samples was lyophilization and pellet pressing.

Proton-induced X-ray emission analysis of atherosclerotic plaques of the carotid bifurcation.

The trace elements of both calcified atherosclerotic plaques and plaque-free vessel walls of the carotid bifurcation from 31 autopsies were investigated using the proton-induced X-ray emission (PIXE) method. The trace elements studied were phosphorus (P), calcium (Ca), chrome (Cr), iron (Fe), copper (Cu), zinc (Zn), lead (Pb), selenium (Se), bromine (Br), strontium (Sr), and rubidium (Rb). All samples contained Fe and Zn. Mercury (Hg) was not detected in any of the samples studied. All plaque-free samples contained Cu and almost all Br and Ca, none Sr. All calcified atherosclerotic plaques contained Ca and almost all Br and Sr. The relative levels of Ca were higher in the calcified plaques than in the plaque-free vessel walls. The relative value of Ca in calcified and uncalcified samples was greatest in the group who had died because of cardiovascular disorders and smallest in the group who had died from other causes. There was a strong positive correlation between the Ca and Sr of the plaque samples and between the P and Br of the plaque-free samples.

Evidence for increased non-ceruloplasmin copper in early-stage human breast cancer serum.

We measured total serum copper and ceruloplasmin levels in pre- and postmenopausal Stage I and II breast cancer (BC) patients and omnivorous and vegetarian controls. The omnivorous groups included 14 premenopausal women [33 +/- 6 (SD) yrs] and 11 postmenopausal women (57 +/- 5 yrs), and the vegetarian groups were comprised of 12 premenopausal subjects (34 +/- 7 yrs) and 11 postmenopausal subjects (59 +/- 5 yrs). There were 13 premenopausal BC patients (39 +/- 7 yrs) and 10 postmenopausal BC patients (66 +/- 6 yrs). Fasting serum samples were taken on three consecutive days, typically four times in the year. Serum ceruloplasmin levels (g/l) were measured by nephelometry utilizing monoclonal antiserum, and total serum copper levels (mumol/l) were determined by proton-induced X-ray emission analysis. Premenopausal patients had higher serum copper levels than their controls (mean 18.7 vs. 16.6, p less than 0.03). For ceruloplasmin, the postmenopausal BC patients had significantly lower levels than pooled postmenopausal controls (0.309 vs. 0.370, p less than 0.001). The copper-to-ceruloplasmin ratio was significantly higher in the pooled cancer groups than in the pooled control groups (3.69 vs. 3.21, p less than 0.001), with similar patterns in both pre- and postmenopausal classes. This high serum copper-to-ceruloplasmin ratio in BC patients may reflect disordered copper metabolism in this disease, which could also have implications for the origin of, or the response to, the cancer process.

Trace element studies on whole human cerebrospinal fluid with external beam PIXE.

External beam PIXE analysis with a 2.4 MeV proton beam was used to determine the concentrations of K, Ca, Fe, Cu, Zn and Br in cerebrospinal fluid from patients having various disorders. The obtained total concentration ranges K 34,000-1,079,000, Ca 5300-81,300, Fe 40-1030, Cu 20-1650, Zn 15-1250 and Br 400-43,000 micrograms/kg are compared with the values given in the literature. In certain patients there were very high CSF bromine levels, but this was shown to be the result of taking medications presented as bromide salts. The possibility of using the method in clinical practice for CSF analysis is considered. The new method of preparing self-supporting films of the samples was used. This method was further optimized by investigating in detail the use of EDTA as a homogenizer.

A new rapid method for quantification of nitrogen in human serum employing the 14N(p,p' gamma)14N reaction: application to human pregnancy.

The total nitrogen concentrations in dried serum from 54 pregnant women and 17 newborn babies were determined by a new application of the 14N(p,p' gamma)14N reaction resonance at 3.9 MeV. The samples were bombarded in a He atmosphere by 4.1 MeV protons from a tandem Van de Graaff accelerator. The mean dry-weight nitrogen concentration in serum sampled during early pregnancy (6-12 weeks; 13.9 g per 100 g +/- 5.4%) was significantly higher than that in serum sampled during late pregnancy (38-42 weeks: 13.0 g per 100 g +/- 3.9%, p less than 0.001) and than that in serum taken from the umbilical cord (13.3 g per 100 g +/- 4.6%, p less than 0.01). The nitrogen levels measured using this rapid nuclear technique, applied for the first time to human serum analysis, agree well with parallel Kjeldahl analyses.

Selenium and other elements in human maternal and umbilical serum, as determined simultaneously by proton-induced X-ray emission.

Using PIXE (proton-induced X-ray emission), we simultaneously determined the concentrations of Se, Ca, Fe, Cu, Zn, Br, and Pb in blood serum from 56 pregnant women, 25 healthy controls, and 31 others with twin pregnancy or some complicating condition (diabetes, hypertension, epilepsy, hepatosis gravidarum, pre-eclampsia, small baby), and in cord-blood serum from 21 newborns. Pellets, pressed from the serum samples after addition of yttrium as an internal standard, mixing, and evaporating at 30 degrees C with or without reduced pressure (less than 1 kPa), were bombarded by 2.2 MeV protons from a Van de Graaff accelerator in the air and the induced X-rays collected by a Ge(Li) detector. Relative to mean Se values for early six- to 12-week pregnancy (0.045 ppm), those for 35-42 week pregnancy (0.028 ppm) were low (p less than 0.001). Umbilical cord blood serum showed even lower values (0.016 ppm, p less than 0.001)--findings in harmony with the incidence pattern of Keshan cardiomyopathy. Pb crossed the placenta; values for cord serum were not significantly different from those in pregnancy serum. Cu, Zn, Fe, and Ca showed the significant expected patterns in the different groups. Compared with the late-pregnancy controls, Fe was high in mothers of small-birth-weight babies (1.70 ppm, p less than 0.02). Br was high in pre-eclampsia (3.59 ppm, p less than 0.05) and mothers with twins (3.61 ppm, p less than 0.05).

In vitro measurement of phosphorus in small bone biopsies using 14 MeV neutron activation analysis.

In studying diseases of bone such as osteoporosis and renal osteodystrophy it is important to have an index of change in bone mineral content. For this purpose we describe a method for measuring the phosphorus content of 100--150 mg bone biopsies. Femoral and iliac crest biopsies from 6 cadavers were each paired with a standard bone of similar size and irradiated with 14 MeV neutrons for 10 minutes. A special rotor ensured the same activating flux across both sample and standard bone which were both wrapped in aluminium foil. The 28Al generated from bone phosphorus (31P(n,alpha)28Al) and the 27Mg generated from the aluminium foils (27Al(n,p)27Mg), which undergo beta-decay have useful gamma rays at 1779 keV and 844 keV respectively. These gamma rays were counted using a 110 cm3 Ge(Li) detector and a rotating frame permitting four 30 s counts of sample and standard alternately (interval for rotation 10 seconds) in identical geometries. Using this system the phosphorus content of the biopsies could be determined with a coefficient of variation of 4.6% using the standard bone as reference and 6% using aluminium foil as reference. In dense bone the P/dry weight ratio was 30--50% greater than in spongy bone. The clinical value of the method is discussed.

Elemental analysis of bone mineral by backscattering of alpha particles.

Backscattering of 2 MeV alpha particles has been applied for the first time to studies of the inorganic substances of human bone. Bone samples taken from the femoral shaft (dense bone) and iliac crest (spongy bone) of 18 cadavers were analysed for the calcium, phosphorus and oxygen content, after lyophilisation and heating for 1.5 h at 500 degrees C to remove water and organic material. The backscattering method produces a spectrum in which all the elements present in the sample except hydrogen can be seen at once. The reproducibility of the measurements was less than 2%. No previous studies were found in the literature of the measurement of elemental oxygen in bone samples. Ca/P ratios varied from 1.97 to 2.47, Ca/O ratios from 0.76 to 0.91 and P/O ratios from 0.34 to 0.41, the latter two being about 10--15% lower than expected if bone mineral were hydroxyapatite (Ca/O = 0.96 and P/O = 0.44). The results are consistent with previous work on Ca/P ratios but the low Ca/O and P/O ratios could not be explained without postulation of crystal or hydroxyl water in bone mineral. Support for the explanation was gained by differential gravimetric and thermal analysis.