Diffusion-weighted magnetic resonance imaging of the fetal brain in intrauterine growth restriction.

OBJECTIVE: Diffusion-weighted magnetic resonance imaging (DWI) is a sensitive method for assessing brain maturation and detecting brain lesions, providing apparent diffusion coefficient (ADC) values as a measure of water diffusion. Abnormal ADC values are seen in ischemic brain lesions, such as those associated with acute or chronic hypoxia. The aim of this study was to assess whether ADC values in the fetal brain were different in fetuses with severe intrauterine growth restriction (IUGR) compared with normal controls. METHODS: Brain magnetic resonance imaging (MRI) with single-shot axial DWI (b = 0 and b = 700 s/mm2 ) was performed in 30 fetuses with severe IUGR (estimated fetal weight < 3rd centile with absent or reversed umbilical artery Doppler flow) and in 24 normal controls of similar gestational age. Brain morphology and biometry were analyzed. ADC values were measured in frontal and occipital white matter, centrum semiovale, thalami, cerebellar hemisphere and pons. Frontal-occipital and frontal-cerebellar ADC ratios were calculated, and values were compared between IUGR fetuses and controls. RESULTS: There was no difference in gestational age at MRI between IUGR and control fetuses (IUGR, 30.2 ± 1.6 weeks vs controls, 30.7 ± 1.4 weeks). Fetal brain morphology and signals were normal in all fetuses. Brain dimensions (supratentorial ± infratentorial) were decreased (Z-score, < -2) in 20 (66.7%) IUGR fetuses. Compared with controls, IUGR fetuses had significantly lower ADC values in frontal white matter (1.97 ± 0.23 vs 2.17 ± 0.22 × 10-3 mm2 /s; P < 0.0001), thalami (1.04 ± 0.15 vs 1.13 ± 0.10 ×10-3 mm2 /s; P = 0.0002), centrum semiovale (1.86 ± 0.22 vs 1.97 ± 0.23 ×10-3 mm2 /s; P = 0.01) and pons (0.85 ± 0.19 vs 0.94 ± 0.12 ×10-3 mm2 /s; P = 0.043). IUGR fetuses had a lower frontal-occipital ADC ratio than did normal fetuses (1.00 ± 0.11 vs 1.08 ± 0.05; P = 0.003). CONCLUSIONS: ADC values in IUGR fetuses were significantly lower than in normal controls in the frontal white matter, thalami, centrum semiovale and pons, suggesting abnormal maturation in these regions. However, the prognostic value of these ADC changes is still unknown. Copyright © 2016 ISUOG. Published by John Wiley & Sons Ltd.

Safety and efficacy of one-stage total laparoscopic treatment of common bile duct stones in children.

BACKGROUND: The purpose of this study is to confirm the effectiveness of total laparoscopic treatment of common bile duct (CBD) stones in children. METHODS: All children who were treated in our department for cholelithiasis were reviewed from 1996 to 2013. Data collection focused on children with CBD stones, including age, sex, symptoms at diagnosis, hepatic and pancreatic blood tests results, US scan results, etiology, detailed surgical technique, operative time, length of hospital stay, complications, and stone-free status or not, at last follow-up. RESULTS: 551 children were treated for cholelithiasis and had undergone laparoscopic cholecystectomy. Among those, 36 children (6.5%) presented with CBD stones with a mean age at symptom onset of 10.4 years (min-max: 4 months-18 years). A majority of the patients presented with hemolytic disease (61%). In 55% of the cases, cholangiography alone or simple serum saline flush of the biliary tree was sufficient to obtain a stone-free CBD. Additional maneuvers with Dormia basket or Fogarty catheter led to 72% of success rate. In 9 cases (25%) of failure of the procedure, 6 patients underwent endoscopic retrograde cholangiopancreatography (ERCP) with endoscopic sphincterotomy (ES), 1 patient was re-operated at day1 for hemorrhage, and 2 patients were followed by US scan with spontaneous evacuation of CBD stones. Mean follow-up was of 2 years (min-max: 1 month-5 years). All patients were stone free at last clinical and radiological evaluation. CONCLUSION: A one-stage total laparoscopic treatment of common bile duct stones in children is a safe, feasible, reproducible, and efficient procedure in 72% of the cases. This rate could be upgraded by a combination of laparoscopic and endoscopic technique during the same anesthesia and preserving Oddi sphincter function. These minimal invasive techniques still need to be developed in children.

Effect of interleukin-2 and selenium on the growth of squamous cell carcinoma cells.

OBJECTIVES: The objectives of this study were to determine whether the expression of the interleukin-2 (IL-2) receptors on squamous cell carcinoma cells can be enhanced in the presence of selenium (Se) and contribute to a greater retardation of tumor growth after locoregional therapy with IL-2. STUDY DESIGN: The growth of the cells was studied after in vitro or dietary supplementation with Se in a murine model. RESULTS: Treatment of established tumors in hosts supplemented with Se with peritumoral injections of IL-2 resulted in 50% reduction of tumor size, whereas treatment of early tumors resulted in 72.4% reduction. The effect was most likely related to a combination of enhanced immune responsiveness and enhanced IL-2 receptor expression on the tumor cells. CONCLUSIONS AND SIGNIFICANCE: The data suggested that local immunotherapy with IL-2 in hosts supplemented with Se may represent an effective modality of treatment for the prevention of recurrences at the site of conventionally treated primary tumors, including tumors that do not express IL-2 receptors.

Prognostic evaluation of Legg-Calvé-Perthes disease by MRI. Part I: the role of physeal involvement.

Twenty-eight unselected hips affected by Legg-Calvé-Perthes disease (LCPD) in 24 patients from 1987 to 1994 had magnetic resonance imaging (MRI) performed. The patients were retrospectively reviewed at a mean follow-up of 5.4 years. An interobserver blind analysis was made to establish the reliability of four MRI parameters: extent of epiphyseal necrosis (EXT), lateral extrusion of the femoral head (LAT), physeal involvement (PHY), and metaphyseal changes (MET). The interobserver analysis resulted in a good reliability for all MRI parameters (concordance, >80%; K index, >0.45). A statistical correlation study (Spearman test) was then done between each MRI parameter and the condition of the hips at follow-up evaluated by Stulberg class and a personal scored system (total score) of clinical-radiographic condition. All MRI parameters appeared well correlated to the Stulberg class and to the total score (S > 0.66; p < 0.05). PHY resulted the strongest correlated parameter (S = 0.84 for Stulberg class; S = 0.91 for total score). Finally our statistical correlation study demonstrates physis involvement to have a high predictive value in LCPD; therefore it can be assumed as the main risk factor in formulating prognosis.

Phosphatidylcholine makes specific activity of the purified Ca(2+)-ATPase from plasma membranes independent of enzyme concentration.

Ca(2+)-ATPase of plasma membranes (PMCA) was isolated from either human or pig red cells by calmodulin-affinity chromatography and supplemented with phosphatidylcholine (PC). The specific activity of the purified PMCA diluted in media with detergent (C(12)E(10)) was very low, and increased with the concentration of the enzyme along a curve that reached the maximum at 8 microg/ml with K(0.5)=1.2-2.5 microg/ml. Such behavior has been described and attributed to self-association of the enzyme (D. Kosk-Kosicka and T. Bzdega, J. Biol. Chem. 263 (1988) 18184-18189). After heat-inactivation, the PMCA was as effective an activator as the intact enzyme, increasing, to the maximum, the specific activity of diluted enzyme with K(0. 5)=2.2 microg/ml. The inactivated PMCA failed to increase the activity of concentrated enzyme, suggesting that activation did not depend on interaction of intact with denatured enzyme molecules. When enough PC was added to the reaction medium to make its final concentration 16-33 microg/ml, the specific activity of the PMCA was maximum and independent of enzyme concentration. Under these conditions, activation by calmodulin lowered to 10%. As a function of the concentration of pure PC, maximum specific activity was reached along a curve with K(0.5)=4 microg/ml. This curve was identical to that of activation at increasing enzyme concentration, suggesting that, in the latter case, activation could have depended on PC contributed to the assay medium by the enzyme. The results show that PC made the purified PMCA solubilized in detergent reach maximum activity at any concentration of the enzyme.

["Classic" vs autolubricant catheterization for endovesicular chemotherapy. Preliminary experience]. / Cateterismo "classico" vs autolubrificante per chemioterapia endovescicale. Esperienza preliminare.

AIM: The aim of this study was to evaluate the frequency of urinary tract infections (UTI) after catheterisation for instillation comparing two systems: the "classic" method and the catheterisation using a new autolubricant device: EasiCath Coloplast. METHODS: During the period of endovesical chemotherapy (between 4 and 48 weeks), 22 patients (6 females and 18 males) were studied, aged between 53 and 78 years old. We have performed 139 instillations using Nelaton Ch 14 or 12 type catheters lubricated with gel based on lidocaine, neomicyn and fluocinolone ("classic" method). Instead 135 patients have been treated with autolubricant devices according to the manufacturer's instructions. After 48 hours from instillation, a total of 274 catheterisation have been examined using urine tests and urine culture with antibiogram. We administered a 5-point visual analogic score to the patients weighing the post-instillation dysuria. RESULTS: With "classic" method UTI frequency is 7.19% (10/139). The most common pathogen has been E. coli (7/10). With autolubricant catheters UTI frequency is 2.96 (4/135). Klebsiella, Enterobacter, as well as E. coli (2/4) have been identified as pathogen. All patients with infections have been treated with targeted antibiotics based on the antibiogram. CONCLUSIONS: We have observed the people with autolubricant catheters left more comfort then those undergoing to the "classic" catheterisation. The frequency of post-catheterisation, dysuria was also reduced. Our data show that the new method is safer and easier to handle then the "classic" one. Moreover, common anaesthetic/antibiotic lubricant have important bacteriostatic effects that reduce the BCG viability.

Juvenile hyaline fibromatosis: a case report.

The authors present a 9-year-old girl with a juvenile hyaline fibromatosis observed before any treatment and after surgical releases. The disorder showed impressive clinical features both concerning the size of the dermal nodules and the degree of limitation of involved joints. Although the follow-up was short, we emphasize the great role surgery can play in reaching very satisfying results, both aesthetically and functionally.

Replacement of Val674 by Pro increases the sensitivity of the plasma membrane Ca2+ pump to inhibition by Mg2+.

A cDNA encoding a plasma membrane Ca2+ pump mutant V674P(ct120) was constructed and expressed in COS-1 cells. Immunoblots of transfected COS-1 membranes showed that the V674P(ct120) and the wild-type hPMCA4b(ct120) proteins were expressed at similar levels. The change of Val674 to Pro reduced the activity of the hPMCA4b(ct120) to an extent similar to that observed previously in the full-length Ca2+ pump (Adamo et al. (1995) J. Biol. Chem. 270, 30111-30114). Despite its lower activity, the apparent affinity for Ca2+ of the V674P(ct120) enzyme was at least as high as that of hPMCA4b(ct120), indicating that substitution of Val674 by Pro did not impair the interaction of the enzyme with Ca2+. The sensitivity of the V674P(ct120) enzyme to inhibition by vanadate was not significantly different from that of the hPMCA4b(ct120), supporting the idea that the mutation did not alter the equilibrium between E2-E1. The study of the Mg2+ dependency of the Ca2+ transport showed that the V674P(ct120) mutant reached maximum activation at 100 microM Mg2+ in contrast with 500 microM in the hPMCA4b(ct120). Furthermore, while at 2 mM Mg2+ the hPMCA4b(ct120) showed no sign of inhibition, the activity of the mutant decreased to less than 50% of the maximum activity observed at 100 microM Mg2+. These results indicate that the decrease in the activity observed upon substitution of Val674 by Pro was due to a higher sensitivity to Mg2+ as inhibitor.

Pre-steady-state kinetic study of the mechanism of inhibition of the plasma membrane Ca(2+)-ATPase by lanthanum.

Lanthanides are known to be effective inhibitors of the PMCa(2+)-ATPase. The effects of LaCl3 on the partial reactions that take place during ATP hydrolysis by the calcium-dependent ATPase from plasma membrane (PMCa(2+)-ATPase) were studied at 37 degrees C on fragmented intact membranes from pig red cells by means of a rapid chemical quenching technique. LaCl3 added before phosphorylation (K0.5 = 2.8 +/- 0.2 microM) raised the kapp of the E2-->E1 transition from 14 +/- 2 to 23 +/- 4 s-1. The effect was independent of Ca2+ and Mg2+, as if La3+ substituted for Mg2+ and/or Ca2+ in accelerating the formation of E1 with higher efficiency. At non-limiting conditions, LaCl3 doubled the apparent concentration of E1 in the enzyme at rest with Ca2+ and Mg2+. LaCl3 during phosphorylation (K0.5 near 20 microM) lowered the vo of the reaction from 300 +/- 20 to 60 +/- 7 pmol/mg of protein/s, a close rate to that in the absence of Mg2+. This effect was reversed by Mg2+ (and not by Ca2+), and the K0.5 for Mg2+ as activator of the phosphorylation reaction increased linearly with the concentration of LaCl3, suggesting that La3+ slowed phosphorylation by displacing Mg2+ from the activation site(s). If added before phosphorylation, LaCl3 lowered the kapp for decomposition of EP to 0.8 +/- 0.1 s-1, a value which is characteristic of phosphoenzyme without Mg2+. The K0.5 for this effect was 0.9 +/- 0.5 microM LaCl3 and increased linearly with the concentration of Mg2+. If added after phosphorylation, LaCl3 did not change the kapp of 90 +/- 7 s-1 of decomposition of EP, suggesting that La3+ displaced Mg2+ from the site whose occupation accelerates the shifting of E1P to E2P. In medium with 0.5 mM MgCl2, 2 microM LaCl3 lowered rapidly the rate of steady-state hydrolysis of ATP by the PMCa(2+)-ATPase to a value close to the rate of decomposition of EP made in medium with LaCl3. Increasing MgCl2 to 10 mM protected the PMCa(2+)-ATPase against inhibition during the first 10 min of incubation. Results show that combination of La3+ to the Mg2+ (and Ca2+) site(s) in the unphosphorylated PMCa(2+)-ATPase accelerates the E2-->E1 transition and inhibits the shifting E1P--> E2P. Since with less apparent affinity La3+ slowed but did not impede phosphorylation, it seems that the sharp slowing of the rate of transformation of E1P into E2P by displacement of Mg2+ was the cause of the high-affinity inhibition of the PMCa(2+)-ATPase by La3+.

Deletion of amino acid residues 18-75 inactivates the plasma membrane Ca2+ pump.

A mutant of the plasma membrane Ca2+ pump hPMCA4b(d18-75)(ct120) containing a deletion of the N-terminal amino acid residues 18-75 and lacking the C-terminal 120 amino acid residues was expressed in COS-1 cells. The deletion in the N-terminal region did not significantly affect the level of expression of the Ca2+ pump. Tryptic digestion of the hPMCA4b(d18-75)(ct120) mutant resulted in the appearance of the same fragments obtained by proteolysis of the hPMCA4b(ct120) enzyme, suggesting that deletion of residues 18-75 neither impeded the insertion in the membrane nor extensively affected the folding of the mutant protein. The functional competence of the hPMCA4b(d18-75)(ct120) enzyme was examined by measuring the Ca2+ transport and the Ca2+ ATPase activity of COS-1 cell microsomes expressing the mutant protein. Both tests proved the mutant to be inactive. Under conditions in which hPMCA4b(ct120) becomes phosphorylated, hPMCA4b(d18-75)(ct120) was incapable of reacting with ATP and Ca2+ to form the phosphoenzyme. Taken together these results suggest that the segment of amino acids 18-75 is essential for the activity of the plasma membrane Ca2+ pump.

Eosinophilic granuloma of C4 causing spinal cord compression.

A rare case of vertebral eosinophilic granuloma (C4) causing spinal cord compression is reported. The clinical, histological and radiological features of this pathological entity are discussed. After surgery a complete neurological recovery was observed. The value, in selected cases, of surgical treatment with total removal of the tumour and reconstruction of the spine to ensure spinal stability and to prevent irreversible neurological deficit is emphasized.

Pre-steady-state kinetic study of the effects of K+ on the partial reactions of the catalytic cycle of the plasma membrane Ca(2+)-ATPase.

The effects of 100 mM K+ on the partial reactions that take place during ATP hydrolysis on the calcium ion-dependent ATPase from plasma membrane (PM-Ca(2+)-ATPase) were studied at 37 degrees C on fragmented intact membranes from pig red cells by means of a rapid chemical quenching technique. At 10 microM [gamma-32P]ATP plus non-limiting concentrations of Ca2+ and Mg2+, K+ increased the k(app) of formation by 140% to 84 11 s-1 and the steady-state level of phosphoenzyme (EP) by 25% to 3.4 0.17 pmol/mg of protein. If added together with [gamma-32P]ATP at the beginning of phosphorylation, K+ was much less effective than if added earlier, indicating that it did not act on the phosphorylation reaction. Measurements of the E2 --> E1 transition by phosphorylation showed that in medium with Ca2+ and Mg2+, K+ increased the k(app) of the transition by 55% to 14 3 s-1 and the apparent concentration of E1 by 45%, suggesting that this may be the cause of the increased rate of phosphorylation observed in enzyme preincubated with K+. The presence of K+ did not change the slow decay of EP without Mg2+ but activated the decay of EP made with Mg2+, increasing its k(app) by 60% to 91 12 s-1. In contrast with observations made during phosphorylation, if added at the beginning of dephosphorylation K+ was fully effective in favouring decomposition of EP made in medium containing no K+. In the presence of either 3mM ATP or 3 mM ATP plus calmodulin, which activate hydrolysis of CaE2P, the effect of K+ on dephosphorylation was conserved. Because the sites for K+ are intracellular and the concentration of K+ in normal red cells is above 100 mM, the effects described here must be taken into account to describe the catalytic cycle of the PM-Ca(2+)-ATPase under physiological conditions.

New rationale in management of tibial agenesis type II: a maturity review of its functional, psychological, and economic value.

It is now 3 years since we published a new surgical approach to type II tibial agenesis. All three patients have reached the end of growth, and we have been able to conduct a maturity review of the functional, psychological, and economic value of this method of treatment. All three patients were very satisfied with the treatment they had received, although it was long and often arduous. We have attempted to compare the costs of primary amputation without complication in the first year of life, followed by supply of at least 10 prostheses, until age 18 years, with that of the reconstructive technique, which is liable to require five operations and considerable periods of inpatients and outpatient care until age of 18 years.

Gluteoperineal gangrene and sciatic nerve palsy after umbilical vessel injection.

Thirteen patients with gluteoperineal necrosis of the skin and subcutaneous tissue without muscle involvement associated with palsy of the peroneal branch of the sciatic nerve were examined at the authors' institution. This syndrome was a complication after umbilical vessel catheterization for injection of medications mainly for respiratory distress and apnea. Skin and subcutaneous damaged tissues required debridement without sedation or anesthesia. Local antibiotic application and ultraviolet rays helped heal the lesions, leaving a star-like scar that did not cause cosmetic disfigurement or fistulae formation. For the residual peroneal nerve palsy with involvement of peroneal, toe extensor, and anterior tibialis musculature, the therapeutic approach initially consisted of physical therapy to the involved muscles of the foot. Surgery was required only to correct the deformity of the foot secondary to muscular deficits. In this series, the final results of the muscles involved secondary to nerve palsy were good in 4 patients, fair in 6, and poor in 3. No excellent result was noted.

Effects of magnesium plus vanadate on partial reactions of the Ca(2+)-ATPase from human red cell membranes.

Under conditions in which pretreatment with Mg2+ plus vanadate activate the Ca(2+)-ATPase, the initial rate of phosphorylation of the enzyme increased from 141 to 259 pmol/mg protein per s while the steady-state level of phosphoenzyme lowered from 1.9 to 1.1 pmol/mg protein. The drop in phosphoenzyme level was caused by incubation and washing during treatment rather than by vanadate. The data allowed to estimate a turnover number for the enzyme that raised by 170% after pretreatment. The results show that the activation of the Ca(2+)-ATPase by Mg2+ plus vanadate is due to changes in the kinetic properties of the enzyme.