Effects of milk replacer acidification and free-access feeding on early life feeding, oral, and lying behavior of dairy calves.

Acidification is a practical way of preserving the bacteriological quality of milk so that it can be fed to calves under free-access conditions. The objectives of this study were to evaluate how milk replacer acidification and free-access feeding affect dairy calf behavior during the first week of life. Sixteen Holstein male calves were purchased at birth and transported to the University of Guelph Kemptville Campus Dairy Education and Research Centre. Calves were randomly assigned to 1 of 4 milk feeding programs: (1) free-access (ad libitum) feeding of acidified milk replacer (22% crude protein and 17% fat, 150 g/L; FA); (2) restricted (6 L/d, 150 g/L) feeding of acidified milk replacer (RA); (3) free-access feeding of nonacidified milk replacer (FN); and (4) restricted feeding of nonacidified milk replacer (RN). Formic acid was used to acidify milk replacer to a target pH between 4.0 and 4.5. Video recordings of each calf at 1, 2, and 6 d were analyzed continuously over 24 h for all occurrences of each behavior in the ethogram. Feeding behavior observations were organized into sucking bouts, from which feeding behavior outcome variables were calculated. Calves consuming acidified milk replacer demonstrated more fragmented feeding patterns, characterized by more pauses within a sucking bout (FA, FN, RA, and RN calves = 12.4, 4.4, 13.7, and 11.9 pauses/bout, respectively) and longer sucking bout duration (FA, FN, RA, and RN calves = 8.8, 5.2, 9.3, and 8.1 min/bout, respectively), than calves fed nonacidified milk replacer. Restricted-fed calves tended to have longer sucking bouts and performed more within-bout sucks (FA, FN, RA, and RN calves = 10.7, 5.8, 13.5, and 14.1, respectively) and pauses than free-access calves. Acidification and free-access feeding did not affect lying duration. Calves assigned to the acidified feeding treatments tended to perform more grooming behavior than those fed nonacidified milk replacer (FA, FN, RA, and RN calves = 0.9, 0.5, 0.8, and 0.6 h/d, respectively). Free-access feeding did not affect grooming duration. The observed differences in feeding and grooming behavior suggest that acidification to a pH between 4.0 and 4.5 may have altered the palatability of milk replacer. Calves assigned to the acidified milk replacer feeding treatments did not, however, show avoidance toward this feedstuff during the first week of life.

Clinical trial on the effects of a free-access acidified milk replacer feeding program on the health and growth of dairy replacement heifers and veal calves.

The objectives of this study were to evaluate the effects of free-access acidified milk replacer feeding on the pre- and postweaning health of dairy and veal calves. Individually housed calves were systematically assigned at birth to 1 of 2 feeding programs: free-access feeding (ad libitum) of acidified milk replacer (ACD, n=249) or traditional restricted feeding (3L fed twice daily) of milk replacer (RES, n=249). Calves were fed milk replacer containing 24% crude protein and 18% fat. Acidified milk replacer was prepared to a target pH between 4.0 and 4.5 using formic acid. Calves were weaned off milk replacer at approximately 6wk of age. Weaning occurred over 5d, and during this weaning period, ACD calves had access to milk replacer for 12h/d and RES calves were offered only one feeding of milk replacer (3 L) daily. Calves were monitored daily for signs of disease. Fecal consistency scores were assigned each week from birth until weaning. A subset of calves was systematically selected for fecal sampling at 3 time points between 7 and 27d of age. Fecal samples were analyzed for enterotoxigenic Escherichia coli F5, Cryptosporidium parvum, rotavirus, and coronavirus. Hip width, hip height, body length, heart girth, and body weight were measured at birth and weaning. Postweaning body weight measurements were collected from the heifers at approximately 8mo of age. Postweaning body weight and carcass grading information was collected from the veal calves at slaughter once a live weight between 300 and 350kg had been achieved. The odds of ACD calves being treated for a preweaning disease event tended to be lower than that of the RES calves (1.2 vs. 5.2%, respectively). Preweaning mortality, postweaning disease treatment, and postweaning mortality did not differ between feeding treatments. The ACD feeding treatment supported greater preweaning average daily gain (0.59 vs. 0.43kg/d) and structural growth than RES feeding. Postweaning average daily gain and carcass characteristics were similar for ACD and RES calves. These results indicate that free-access acidified milk replacer feeding tended to support improved health, and greater body weight gain and structural growth during the preweaning period; these effects did not persist in the postweaning period. The growth advantage observed before weaning in the ACD calves likely disappeared due to the weaning methods used.

Spectral CT of carotid atherosclerotic plaque: comparison with histology.

OBJECTIVE: To distinguish components of vulnerable atherosclerotic plaque by imaging their energy response using spectral CT and comparing images with histology. METHODS: After spectroscopic calibration using phantoms of plaque surrogates, excised human carotid atherosclerotic plaques were imaged using MARS CT using a photon-processing detector with a silicon sensor layer and microfocus X-ray tube (50 kVp, 0.5 mA) at 38-µm voxel size. The plaques were imaged, sectioned and re-imaged using four threshold energies: 10, 16, 22 and 28 keV; then sequentially stained with modified Von Kossa, Perl's Prussian blue and Oil-Red O, and photographed. Relative Hounsfield units across the energies were entered into a linear algebraic material decomposition model to identify the unknown plaque components. RESULTS: Lipid, calcium, iron and water-like components of plaque have distinguishable energy responses to X-ray, visible on spectral CT images. CT images of the plaque surface correlated very well with histological photographs. Calcium deposits (>1,000 µm) in plaque are larger than iron deposits (<100 µm), but could not be distinguished from each other within the same voxel using the energy range available. CONCLUSIONS: Spectral CT displays energy information in image form at high spatial resolution, enhancing the intrinsic contrast of lipid, calcium and iron within atheroma. KEY POINTS: Spectral computed tomography offers new insights into tissue characterisation. Components of vulnerable atherosclerotic plaque are spectrally distinct with intrinsic contrast. Spectral CT of excised atherosclerotic plaques can display iron, calcium and lipid. Calcium deposits are larger than iron deposits in atheroma. Spectral CT may help in the non-invasive detection of vulnerable plaques.

Spectroscopic (multi-energy) CT distinguishes iodine and barium contrast material in MICE.

OBJECTIVE: Spectral CT differs from dual-energy CT by using a conventional X-ray tube and a photon-counting detector. We wished to produce 3D spectroscopic images of mice that distinguished calcium, iodine and barium. METHODS: We developed a desktop spectral CT, dubbed MARS, based around the Medipix2 photon-counting energy-discriminating detector. The single conventional X-ray tube operated at constant voltage (75 kVp) and constant current (150 microA). We anaesthetised with ketamine six black mice (C57BL/6). We introduced iodinated contrast material and barium sulphate into the vascular system, alimentary tract and respiratory tract as we euthanised them. The mice were preserved in resin and imaged at four detector energy levels from 12 keV to 42 keV to include the K-edges of iodine (33.0 keV) and barium (37.4 keV). Principal component analysis was applied to reconstructed images to identify components with independent energy response, then displayed in 2D and 3D. RESULTS: Iodinated and barium contrast material was spectrally distinct from soft tissue and bone in all six mice. Calcium, iodine and barium were displayed as separate channels on 3D colour images at <55 microm isotropic voxels. CONCLUSION: Spectral CT distinguishes contrast agents with K-edges only 4 keV apart. Multi-contrast imaging and molecular CT are potential future applications.

Reproducibility of sonographic assessment of fetal liver length in diabetic pregnancies.

OBJECTIVES: Assessing fetal liver size might be useful in monitoring the effectiveness of the treatment of diabetes in pregnancy. We aimed to assess the reproducibility of fetal liver-length measurement in pregnant women with diabetes. METHODS: From 3 April 2006 to 5 December 2006, we assessed intraobserver and interobserver variation of fetal liver-length measurements on 55 sonograms in 44 pregnant women with diabetes, 34 of whom had gestational diabetes. The mean maternal age was 33 years, the mean maternal weight was 92 kg and the mean body mass index (BMI) was 33.7 kg/m(-2). The effect of covariates BMI, gestational age and maternal age on the reproducibility of fetal liver length was assessed by calculating intraobserver SD ratios. We compared liver length with abdominal circumference and gestational age. Nine of 12 sonographers scanned, on average, six women (range, 3-12) as the first sonographer, and all 12 sonographers scanned, on average, four women (range, 1-10) as the second sonographer. The data were analyzed using a hierarchical linear model. RESULTS: Measurement of fetal liver length was reproducible. The intraobserver SD was 3.06 (95% CI, 2.68-3.59) mm; the interobserver SD was 2.17 (95% CI, 0.59-4.83) mm; the intraobserver correlation was 0.77 (95% CI, 0.63-0.87), and the interobserver correlation was 0.84 (95% CI, 0.51-0.99). The covariate effects were minimal, the SD for a 1-unit increase in the covariate was 1.06 for gestational age, 0.98 for BMI, and 0.97 for maternal age. CONCLUSIONS: Measurement of fetal liver length in the diabetic pregnancy is reproducible. It is worthy of further investigation as a parameter for monitoring the effectiveness of treatment of the diabetic pregnancy.

Sonographic estimation of fetal weight: comparison of bias, precision and consistency using 12 different formulae.

OBJECTIVES: To determine the major sources of error in ultrasonographic assessment of fetal weight and whether they have changed over the last decade. METHODS: We performed a prospective observational study in 1991 and again in 2000 of a mixed-risk pregnancy population, estimating fetal weight within 7 days of delivery. In 1991, the Rose and McCallum formula was used for 72 deliveries. Inter- and intraobserver agreement was assessed within this group. Bland-Altman measures of agreement from log data were calculated as ratios. We repeated the study in 2000 in 208 consecutive deliveries, comparing predicted and actual weights for 12 published equations using Bland-Altman and percentage error methods. We compared bias (mean percentage error), precision (SD percentage error), and their consistency across the weight ranges. RESULTS: 95% limits of agreement ranged from - 4.4% to + 3.3% for inter- and intraobserver estimates, but were - 18.0% to 24.0% for estimated and actual birth weight. There was no improvement in accuracy between 1991 and 2000. In 2000 only six of the 12 published formulae had overall bias within 7% and precision within 15%. There was greater bias and poorer precision in nearly all equations if the birth weight was < 1,000 g. CONCLUSIONS: Observer error is a relatively minor component of the error in estimating fetal weight; error due to the equation is a larger source of error. Improvements in ultrasound technology have not improved the accuracy of estimating fetal weight. Comparison of methods of estimating fetal weight requires statistical methods that can separate out bias, precision and consistency. Estimating fetal weight in the very low birth weight infant is subject to much greater error than it is in larger babies.

The ultrasound halo sign in angiolymphoid hyperplasia of the temporal artery.

An ultrasound halo sign surrounding the temporal artery is a well recognized feature associated with giant cell arteritis. We report a previously unreported case of this halo sign being present around the temporal artery due to angiolymphoid hyperplasia with eosinophilia (ALHE) in a young female patient.

Normal obstetric ultrasound reduces the risk of Down syndrome in fetuses of older mothers.

The objective of this study is to determine whether a normal fetal morphology ultrasound scan in women older than 35 years reduces the risk of aneuploidy. We reviewed the results of amniocentesis and second trimester sonogram in all women older than 35 years from 1991 to 1995. None had prior screening. We excluded fetuses with structural anomalies. We determined the sensitivity and specificity of minor markers in detecting Down syndrome and also determined the reduction in risk of a normal sonogram. Among the 2060 women older than 35 years giving birth during the study period, 16 (0.78%) delivered an infant with Down syndrome. Of the 16 fetuses, two had no prenatal testing or ultrasound, two had invasive testing but no second trimester sonogram, five had a normal sonogram and seven had one or more sonographic markers of Down syndrome. At least 17% of women older than 35 years did not participate in prenatal testing or ultrasound. Ultrasound detected Down syndrome with a sensitivity of 59% (95% confidence interval: 45-72%), a false-positive rate of 10.6% (9.4-11.8%) and a positive predictor value of 1 in 9. The likelihood of having normal karyotype if the sonogram was normal was 0.46 (0.31-0.61). In women older than 35 years, a normal second trimester sonogram reduces the risk of Down syndrome by more than 50%. At least 17% of women older than 35 years do not participate in prenatal testing or ultrasound.

Intrinsic measurement bias on computed tomography scout view is unpredictable: Computed tomography pelvimetry using a phantom.

Our aim was to determine the degree of bias in CT scanogram measurements. We obtained standard lateral and anteroposterior (AP) pelvimetry scanograms of a phantom pelvis after placing ball bearings or aluminium rods to mark bony landmarks. Computed tomography pelvimetry was carried out at the manufacturer-recommended table height on two commercial CT scanners and at 10-mm increments up to 50 mm above and below this height. The AP inlet, AP outlet, interspinous distance and transverse diameters were each measured three times for each scanogram. The true measurements were obtained directly from the disassembled phantom. Bias was defined as the difference between the CT measurement and the true measurement. Observer error was negligible. The transverse diameter was overestimated at high table positions and underestimated at low table positions on both scanners (+6 to -10 mm). After correcting for geometric distortion, up to 6 mm bias was still present. The point at which no bias occurred was different for each scanner and did not correspond to the manufacturers' recommended table height. The outlet was overestimated on both scanners by up to 5 mm. The true inlet measurement was overestimated by 1.2 mm. The interspinous distance was minimally underestimated on both scanners. The measurements on CT scanogram were underestimated or overestimated in an inconsistent and unpredictable fashion, varying from one type of measurement to another and from CT scanner to CT scanner. This has implications for the accuracy and clinical utility of measurements obtained from a CT scanogram.

Autocrine signalling through erbB receptors promotes constitutive activation of protein kinase B/Akt in breast cancer cell lines.

The protein kinase PKB/Akt plays a pivotal role in promoting cell survival and proliferation. This study investigated the regulation of PKB/Akt activity in breast cancer cells. In primary invasive breast cancers PKB/Akt exhibited elevated phosphorylation at regulatory site Ser473 in 80% of cases, using immunohistochemistry. The degree of phospho-PKB/Akt immunoreactivity was positively correlated with the extent of its nuclear accumulation. Moderate/strong staining was seen in 31% of the samples but was absent in tumour-associated normal breast epithelia. To examine the mechanisms of PKB/Akt activation, we studied its phosphorylation in a panel of breast cancer cell lines. PKB/Akt was constitutively phosphorylated on both regulatory sites (Thr308 and Ser473) in the absence of serum growth factors in 7 of 8 lines but not in two cell lines derived from normal breast epithelia. Further analysis revealed that constitutive PKB/Akt phosphorylation was associated with loss of PTEN phosphatase expression (CAL51, MDA-MB-468, BT549 cells) and constitutive activation of erbB2 (SKBR3, BT474 cells). In two further breast cancer lines (T47D and HS578T) PKB/Akt phosphorylation was dependent upon autocrine factors acting primary through the epidermal growth factor receptor (EGFR) and erbB2. Conditioned medium from HS578T cells stimulated EGFR-dependent PKB/Akt phosphorylation in normal breast cells. These results demonstrate that PKB/Akt is frequently activated in breast cancer through diverse mechanisms, including autocrine signalling via erbB receptors.

Protein kinase C-epsilon promotes adipogenic commitment and is essential for terminal differentiation of 3T3-F442A preadipocytes.

The role of protein kinase C (PKC) isoforms in the commitment of multipotent fibroblasts to the adipocyte lineage and in their terminal differentiation into mature adipocytes was investigated. Ectopic overexpression of PKC-epsilon, but not other PKC isoforms, committed multipotent NIH-3T3 cells to adipogenic differentiation in the presence of hormonal inducers. In committed 3T3-F442A preadipocytes, PKC-epsilon protein expression increased during the course of terminal differentiation and cell-permeable PKC-epsilon inhibitory peptides, which prevent interaction with RACK (receptor for activated C-kinase) proteins, severely inhibited differentiation. PKC-epsilon accumulated in the nuclei of 3T3-F442A cells shortly after induction of differentiation and exhibited a distinctive punctate speckling immunocytochemical staining pattern. The spatiotemporal aspects of PKC-epsilon localization and expression coincided with that of C/EBP-beta, a transcription factor critically involved in promoting the early phase of adipogenesis. Collectively, these results demonstrate a role for PKC-epsilon in both adipogenic commitment and preadipocyte terminal differentiation.

The isoform-specific regulation of apoptosis by protein kinase C.

The process of apoptosis is regulated at several levels through phosphorylation by many different protein kinases. The protein kinase C (PKC) family, which comprises at least 10 isoforms with distinct means of regulation and tissue distribution patterns, have been shown to exert both inhibitory and stimulatory influences on apoptosis. This review details recent progress made in determining the roles played by individual PKC isoforms in the control of apoptosis, with reference to their target substrates and actions in different cell types. Although notable exceptions exist, the weight of evidence indicates that the alpha, beta, epsilon and atypical isoforms are anti-apoptotic in their action, whereas the delta and theta isoforms are usually involved in the promotion of apoptosis.

The parathyroid hormone-related protein receptor is expressed in breast cancer bone metastases and promotes autocrine proliferation in breast carcinoma cells.

Overproduction of parathyroid hormone-related protein (PTHRP) occurs in a high proportion of primary breast cancers (PBC) and is strongly implicated in their metastatic spread to bone. Although the PTHRP-receptor (PTHRP-R) is often coexpressed with PTHRP in PBC, its role in regulating breast cancer cell proliferation and metastases to bone remains unclear. The aims of this study were to determine the expression of the PTHRP-R in breast cancer bone metastases (BM) and to investigate the effects of PTHRP-R overexpression on breast cancer cell proliferation. PTHRP-R expression occurred in 85% (11 out of 13) of BM compared with 58% (39 out of 67) of PBC. Median expression was higher (P<0.05) in BM compared with PBC. PTHRP increased cAMP accumulation and DNA synthesis in MCF-7 cells stably overexpressing the PTHRP-R (MCF-7(WTR)) but not in MCF-7(VEC) control cells. The increase in DNA synthesis was mimicked by the cAMP pathway activator forskolin. The receptor antagonist PTHRP(7-34) reduced DNA synthesis in MCF-7(WTR) cells, but not MCF-7(VEC) cells, indicating that receptor overexpression promotes autocrine PTHRP activity. MCF-7(WTR) cells showed increased mitogenic responsiveness to fetal calf serum and reduced doubling times. PTHRP induced weak activation of ERK1 and ERK2 and potentiated their activation by serum growth factors. Collectively these results show that the PTHRP-R is frequently expressed in breast cancer BM and indicate that receptor overexpression drives proliferation via autocrine signals that are mediated via cAMP and ERK pathways.

ZD1839 (Iressa), a novel epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, potently inhibits the growth of EGFR-positive cancer cell lines with or without erbB2 overexpression.

Overexpression of the growth factor receptors EGFR and erbB2 occurs frequently in several human cancers and is associated with aggressive tumour behaviour and poor patient prognosis. We have investigated the effects of ZD1839 (Iressa), a novel EGFR tyrosine kinase inhibitor, on the growth, in vitro and in vivo, of human cancer cell lines expressing various levels of EGFR and erbB2. Proliferation of EGFR-overexpressing A431 and MDA-MB-231 cells in vitro was potently inhibited (50%-70%) by ZD1839 with half-maximally effective doses in the low nanomolar range. In parallel, ZD1839 blocked autophosphorylation of EGFR and prevented activation of PLC-gamma 1, ERK MAP kinases and PKB/Akt by EGF. It also inhibited proliferation in EGFR(+) cancer cell lines overexpressing erbB2 (SKBr3, SKOV3, BT474) by between 20% and 80%, effects which correlated with inhibition of EGF-dependent erbB2 phosphorylation and activation of ERK MAP kinase and PKB/Akt in SKOV3 cells. Oral administration of ZD1839 inhibited the growth of MDA-MB-231 and SKOV3 tumours, established as xenografts in athymic mice, by 71% and 32%, respectively. Growth inhibition coincided with reduced proliferation but no change in apoptotic index. Collectively, these results show that ZD1839, at the doses studied, is a potent inhibitor of proliferation not only in cells overexpressing EGFR but also in EGFR(+) cells that overexpress erbB2.

Back to the future: the human protein index (HPI) and the agenda for post-proteomic biology.

The effort to produce an index of all human proteins (the human protein index, or HPI) began twenty years ago, before the initiation of the human genome program. Because DNA sequencing technology is inherently simpler and more scalable than protein analytical technology, and because the finiteness of genomes invited a spirit of rapid conquest, the notion of genome sequencing has displaced that of protein databases in the minds of most molecular biologists for the last decade. However, now that the human genome sequence is nearing completion, a major realignment is under way that brings proteins back to the center of biological thinking. Using an influx of new and improved protein technologies--from mass spectrometry to re-engineered two-dimensional (2-D) gel systems, the original objectives of the HPI have been expanded and the time frame for its execution radically shortened. Several additional large scale technology efforts flowing from the HPI are also described.

Studies of epidermal growth factor receptor inhibition in breast cancer.

Until recently, there has been little knowledge on the growth control of oestrogen receptor (ER)-negative ductal carcinoma in situ (DCIS) and invasive breast cancer. The recent development of DCIS models, such as transgenic mice, cell-line xenograft models and, importantly, in vivo human DCIS xenograft models has facilitated the investigation and understanding of the control of growth of early pre-invasive breast lesions. Recent studies have shown that ER-negative DCIS, unlike ER-positive DCIS, is hormone independent and does not respond to anti-oestrogen treatment. Moreover, DCIS of the comedo type utilises type I tyrosine kinase growth factors, such as epidermal growth factor receptor (EGFR) and c-erbB-2, in receptor signalling for growth. New data underscore the importance of EGFR as the major modulating growth factor receptor in the control of proliferation in the breast. Pre-clinical studies performed on human DCIS xenografts in nude mice suggest a potential role for EGFR tyrosine kinase inhibitors (EGFR-TKIs). More specifically, ZD1839, a novel orally active and selective EGFR-TKI, has been shown to produce a response in DCIS through a decrease in epithelial proliferation. These findings have enhanced our knowledge of signal transduction pathways in cancer and indicate that tyrosine kinase blockade of EGFR has potential for the treatment and chemoprevention of DCIS. It is hoped that further advances in this area and evaluation of EGFR-TKIs in Phase II/III clinical trials will allow their therapeutic potential as anticancer agents to be appreciated.

Outcome of primary vesicoureteric reflux detected following fetal renal pelvic dilatation.

OBJECTIVE: Postnatal investigation of mild degrees of fetal hydronephrosis has allowed subsequent detection of infants with vesicoureteric reflux (VUR). This study was designed to provide short to medium term information on such infants who had primary VUR, the rates of renal damage and progression over time, the risk factors for such damage and to compare the characteristics of those who had mild dilatation of the fetal renal pelvis (4-9 mm) with those who had moderate-severe dilatation (> or = 10 mm). METHODOLOGY: Since June 1989, infants whose antenatal sonography had identified a fetal renal pelvis with an anteroposterior diameter of > 4 mm were investigated postnatally with renal ultrasonography and micturating cystourethrogram (MCU), and placed on antimicrobial prophylaxis. Those with VUR received 99mTc-dimercaptosuccinic acid (DMSA) scintigraphy. Infants were followed until discharge based on resolution of VUR, surgery, or low grade VUR. A 5.5 year cohort between June 1989 and December 1994 formed the study population. A review of notes and clinical review (if still under follow up) was undertaken. Vesicoureteric reflux on MCU was regraded according to the International Classification, and reflux nephropathy on DMSA scans was regraded according to criteria proposed by Goldraich. Regression analysis was used to assess risk factors for renal damage. RESULTS: There were 69 infants (37 girls, 32 boys) who were identified with primary VUR, with 37/69 having bilateral reflux. Eight had a urinary tract infection during the follow-up period. There was a broad distribution of grades of reflux detected (Grades I-3, Grades II-23, Grades III-19, Grades IV - 17, Grades V-7). 99m-Tc-dimercaptosuccinic acid scans on 57/69 (83%) demonstrated renal damage in eight infants (14%). This was predominantly global contraction of function. No progression of renal damage was seen over 2-7 years. Regression analysis showed a strong association between Grades IV, V reflux and the presence of renal damage (P < 0.001). Review of the degrees of fetal renal pelvic dilatation showed that 60/69 infants were detected because of mild (4-9 mm) dilatation. The majority (43/60) had lower grades of reflux (Grades I, II, 3), but there was no obvious cut-off between 4 and 9 mm that could predict high grade VUR (Grades IV, V). CONCLUSIONS: The use of 4 mm to define an abnormal fetal renal pelvis allows a much larger group of infants with high grade primary VUR to be detected than if a higher cut-off measurement is used. Although it also detects many more infants with low grade primary VUR, there is no obvious cut-off point at which this effect predominates. Progressive renal damage was not seen in follow up of up to 7 years of age. Renal damage on DMSA scanning in this group is almost exclusively a pattern of global contraction of function. The presence of high-grade VUR appears to be the only important factor in predicting the presence of renal damage.

Cerebellar vermis diameter at cranial sonography for assessing gestational age in low-birth-weight infants.

BACKGROUND: Clinical assessment of gestational age for very-low-birth-weight infants is often inaccurate. Survival rates are more dependent on gestational age than on the birth weight. OBJECTIVE: To assess whether cerebellar vermis diameter might predict gestational age in infants under 2,000 g and/or under 32 weeks' gestation. MATERIALS AND METHODS: We carried out a retrospective review of the hard-copy images of midline sagittal views of the cerebellum obtained at cranial sonography, performed via the anterior or posterior fontanelle, in 518 infants admitted to a regional neonatal intensive care unit between June 1991 and November 1996. The vermis diameter was measured from the base of the fourth ventricle to the junction of folium and tuber vermis. We generated regression equations for estimating gestational age from vermis diameter, and from vermis diameter and birth weight, for the 86 infants of known gestational age (less than 32 weeks), with birth weight under 2,000 g and who had scans carried out within 1 week of birth. RESULTS: Measurement of cerebellar vermis diameter alone allowed prediction of gestational age to +/- 1.53 weeks using a 68 % prediction interval, or +/- 3.0 weeks using a 95 % prediction interval. Gender was not significant in the regression analysis. CONCLUSION: Cerebellar vermis diameter predicts gestational age with slightly more precision than the new Ballard score.

Growth hormone-dependent differentiation of 3T3-F442A preadipocytes requires Janus kinase/signal transducer and activator of transcription but not mitogen-activated protein kinase or p70 S6 kinase signaling.

The signals mediating growth hormone (GH)-dependent differentiation of 3T3-F442A preadipocytes under serum-free conditions have been studied. GH priming of cells was required before the induction of terminal differentiation by a combination of epidermal growth factor, tri-iodothyronine, and insulin. Cellular depletion of Janus kinase-2 (JAK-2) using antisense oligodeoxynucleotides (ODNs) prevented GH-stimulated JAK-2 and signal transducer and activator of transcription (STAT)-5 tyrosine phosphorylation and severely attenuated the ability of GH to promote differentiation. Although p42(MAPK)/p44(MAPK) mitogen-activated protein kinases were activated during GH priming, treatment of cells with PD 098059, which prevented activation of these kinases, did not block GH priming. However, antisense ODN-mediated depletion of mitogen-activated protein kinases from the cells showed that their expression was necessary for terminal differentiation. Similarly, although p70(s6k) was activated during GH priming, pretreatment of cells with rapamycin, which prevented the activation of p70(s6k), had no effect on GH priming. However, rapamycin did partially block epidermal growth factor, tri-iodothyronine, and insulin-stimulated terminal differentiation. By contrast, cellular depletion of STAT-5 with antisense ODNs completely abolished the ability of GH to promote differentiation. These results indicate that JAK-2, acting specifically via STAT-5, is necessary for GH-dependent differentiation of 3T3-F442A preadipocytes. Activation of p42(MAPK)/p44(MAPK) and p70(s6k) is not essential for the promotion of differentiation by GH, although these signals are required for GH-independent terminal differentiation.