Randomised study showed that recorded maternal voices reduced pain in preterm infants undergoing heel lance procedures in a neonatal intensive care unit.

AIM: Alleviating pain in neonates should be the goal of all caregivers. We evaluated whether recorded maternal voices were safe and effective in limiting pain in preterm infants undergoing heel lance procedures in the neonatal intensive care unit of an Italian children's hospital. METHODS: This prospective, controlled study took place from December 2013 to December 2015. We enrolled 40 preterm infants, born at a 26-34 weeks of gestation, at a corrected gestational age 29-36 weeks and randomised them to listen or not listen to a recording of their mother's voice during a painful, routine heel lance for blood collection. Changes in the infants' Premature Infant Pain Profile, heart rate, oxygen saturation and blood pressure during the procedure were compared by analysis of variance. Possible side effects, of apnoea, bradycardia, seizures and vomiting, were also recorded. RESULTS: Both groups showed a marked increase in PIPP scores and decrease in oxygen saturation during the procedure, but infants in the treatment group had significantly lower PIPP scores (p = 0.00002) and lower decreases in oxygen saturation (p = 0.0283). No significant side effects were observed. CONCLUSION: Using recorded maternal voices to limit pain in preterm infants undergoing heel lance procedures appeared safe and effective.

Tumor MGMT promoter hypermethylation changes over time limit temozolomide efficacy in a phase II trial for metastatic colorectal cancer.

BACKGROUND: Objective response to dacarbazine, the intravenous form of temozolomide (TMZ), in metastatic colorectal cancer (mCRC) is confined to tumors harboring O(6)-methylguanine-DNA-methyltransferase (MGMT) promoter hypermethylation. We conducted a phase II study of TMZ enriched by MGMT hypermethylation in archival tumor (AT), exploring dynamic of this biomarker in baseline tumor (BT) biopsy and plasma (liquid biopsy). PATIENTS AND METHODS: We screened 150 mCRC patients for MGMT hypermethylation with methylation-specific PCR on AT from FFPE specimens. Eligible patients (n = 29) underwent BT biopsy and then received TMZ 200 mg/m(2) days 1-5 q28 until progression. A Fleming single-stage design was used to determine whether progression-free survival (PFS) rate at 12 weeks would be ≥35% [H0 ≤ 15%, type I error = 0.059 (one-sided), power = 0.849]. Exploratory analyses included comparison between MGMT hypermethylation in AT and BT, and MGMT methylation testing by MethylBEAMing in solid (AT, BT) and LB with regard to tumor response. RESULTS: The PFS rate at 12 weeks was 10.3% [90% confidence interval (CI) 2.9-24.6]. Objective response rate was 3.4% (90% CI 0.2-15.3), disease control rate 48.3% (90% CI 32.0-64.8), median OS 6.2 months (95% CI 3.8-7.6), and median PFS 2.6 months (95% CI 1.4-2.7). We observed the absence of MGMT hypermethylation in BT in 62.7% of tumors. CONCLUSION: Treatment of mCRC with TMZ driven by MGMT promoter hypermethylation in AT samples did not provide meaningful PFS rate at 12 weeks. This biomarker changed from AT to BT, indicating that testing BT biopsy or plasma is needed for refined target selection.

Photo-activated raster scanning thermal imaging at sub-diffraction resolution.

Active thermal imaging is a valuable tool for the nondestructive characterization of the morphological properties and the functional state of biological tissues and synthetic materials. However, state-of-the-art techniques do not typically combine the required high spatial resolution over extended fields of view with the quantification of temperature variations. Here, we demonstrate quantitative far-infrared photo-thermal imaging at sub-diffraction resolution over millimeter-sized fields of view. Our approach combines the sample absorption of modulated raster-scanned laser light with the automated localization of the laser-induced temperature variations imaged by a thermal camera. With temperature increments ∼0.5-5 °C, we achieve a six-time gain with respect to our 350-µm diffraction-limited resolution with proof-of-principle experiments on synthetic samples. We finally demonstrate the biological relevance of sub-diffraction thermal imaging by retrieving temperature-based super-resolution maps of the distribution of Prussian blue nanocubes across explanted murine skin biopsies.

Protonation and conformational dynamics of GFP mutants by two-photon excitation fluorescence correlation spectroscopy.

GFP mutants are known to display fluorescence flickering, a process that occurs in a wide time range. Because serine 65, threonine 203, glutamate 222, and histidine 148 have been indicated as key residues in determining the GFP fluorescence photodynamics, we have focused here on the role of histidine 148 and glutamate 222 by studying the fluorescence dynamics of GFPmut2 (S65A, V68L, and S72A GFP) and its H148G (Mut2G) and E222Q (Mut2Q) mutants. Two relaxation components are found in the fluorescence autocorrelation functions of GFPmut2: a 10-100 micros pH-dependent component and a 100-500 micros laser-power-dependent component. The comparison of these three mutants shows that the mutation of histidine 148 to glycine induces a 3-fold increase in the protonation rate, thereby indicating that the protonation-deprotonation of the chromophore occurs via a proton exchange with the solution mediated by the histidine 148 residue. The power-dependent but pH-independent relaxation mode, which is not affected by the E222Q and H148G mutations, is due to an excited-state process that is probably related to conformational rearrangements of the chromophore after the photoexcitation, more than to the chromophore excited-state proton transfer.

Human colostrum T lymphocytes and their effector cytokines actively aid the development of the newborn immune system.

Colostrum contains soluble and cellular components, the latter mainly T lymphocytes. We expanded in vitro colostrum T lymphocytes (CoTL) to evaluate phenotype and capability of cytokine production. We also considered paired cord blood T-lymphocytes (CBTL) representing the newborn "virgin" immune system. CoTL showed memory phenotype while CBTL expressed mainly naïve phenotype. CoTL included a balanced percentage of helper and cytotoxic subsets. We observed higher percentages of IL-2 (p=0.003) and IL-4 (p=0.027) producing cells by helper rather than by cytotoxic T lymphocytes. The greatest percentage of IFN-gamma producing cells was in cytotoxic cells (p=0.0048), while no difference was found for IL-10. Cord blood samples consisted of a statistically significant greater percentage of helper than cytotoxic cells (p<0.001), with a low percentage of cytokine producing cells, confirming the immaturity of the newborns immune system. CBTL percentage of IL-2 producing cells was higher for helper than cytotoxic subset (p<0.001). We observed a greater percentage of IFN-gamma (p=0.001), IL-4 (p=0.003) and IL-10 (p<0.001) producing cells by cytotoxic than helper T lymphocytes. CoTL demonstrated to protect the newborn through the mothers previous immune experience and to supply active cytokines, which can help the postnatal development of both T type 1/T type 2 response.

Severe neonatal renal failure after maternal use of angiotensin II type I receptor antagonists.

Sartans are selective type 1 angiotensin II receptor antagonists that are used for treatment of arterial hypertension. We report a case of severe renal failure required dialysis after the use of olmesartan in the last month of pregnancy. Exposure to sartans during the last period of gestation seems to be associated with high risk of congenital malformations. It is important to stress that the use of these drugs during pregnancy must be avoided, especially in the third trimester.

Author Correction: µMAPPS: a novel phasor approach to second harmonic analysis for in vitro-in vivo investigation of collagen microstructure.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Hexavalent vaccines: characteristics of available products and practical considerations from a panel of Italian experts.

Combination vaccines represent a valuable technological innovation in the field of infectious disease prevention and public health, because of their great health and economic value from the individual, societal, and healthcare system perspectives. In order to increase parents' and healthcare professionals' confidence in the vaccination programs and maintain their benefits to society, more information about the benefits of innovative vaccination tools such as combination vaccines is needed. Purpose of this work is an examination of available hexavalent vaccines, that protect against Diphtheria, Tetanus, Pertussis, Poliomyelitis, Hepatitis B and Haemophilus influenzae type b infections. From the epidemiological updates of vaccine preventable diseases to the vaccine development cycle, from the immunogenicity of antigenic components to the safety and co-administration with other vaccines, several aspects of available hexavalent vaccines are discussed and deepened. Also a number of practical considerations on schedules, age of employment, strategies for vaccination recovery, vaccination in at-risk births are issued, based on the recommendations of Italian Ministry of Health, Italian Society of Pharmacology (SIF), Italian Society for Pediatrics (SIP), Italian Federation of Family Paediatricians (FIMP) and Italian Society of Hygiene, Preventive Medicine and Public Health (SItI).

Outcome of infants from mothers with anti-SSA/Ro antibodies.

OBJECTIVE: To evaluate the incidence of electrocardiographic and laboratory abnormalities in neonates born from mothers with connective tissue disease and positive for anti-SSA/Ro antibodies. STUDY DESIGN: Electrocardiogram, blood cell counts, liver and renal function tests prospectively obtained from 51 infants born from anti-SSA/Ro-positive mothers with connective tissue disease were compared with those obtained from 50 control infants born from mothers with anti-extractable nuclear antigen (ENA)-negative connective tissue disease. One infant with congenital complete heart block was excluded from analysis. RESULTS: No infant showed sinus bradycardia. A first-degree atrioventricular block at birth was observed in five study group and no control group infants, P=0.023. Atrioventricular blocks spontaneously reverted or remained stable during the first year of life. Mean corrected QT value of infants born from anti-SSA/Ro-positive mothers was slightly prolonged as compared with the control group (0.404+/-0.03 s vs 0.395+/-0.02 s; P=0.060). CONCLUSIONS: Infants exposed to anti-SSA/Ro antibodies had a significantly higher prevalence of first-degree atrioventricular block. At variance with previous studies, we observed a low frequency of hematologic abnormalities and no cases of hepatobiliary disease.

µMAPPS: a novel phasor approach to second harmonic analysis for in vitro-in vivo investigation of collagen microstructure.

Second Harmonic Generation (SHG) is a label-free imaging method used to monitor collagen organization in tissues. Due to its sensitivity to the incident polarization, it provides microstructural information otherwise unreachable by other intensity based imaging methods. We develop and test a Microscopic Multiparametric Analysis by Phasor projection of Polarization-dependent SHG (µMAPPS) that maps the features of the collagen architecture in tissues at the micrometer scale. µMAPPS retrieves pixel-by-pixel the collagen fibrils anisotropy and orientation by operating directly on two coupled phasor spaces, avoiding direct fitting of the polarization dependent SHG signal. We apply µMAPPS to fixed tissue sections and to the study of the collagen microscopic organization in tumors ex-vivo and in-vivo. We develop a clustering algorithm to automatically group pixels with similar microstructural features. µMAPPS can perform fast analyses of tissues and opens to future applications for in-situ diagnosis of pathologies and diseases that could assist histo-pathological evaluation.

A randomised, double blind, placebo controlled trial of the effect of theophylline in prevention of vasomotor nephropathy in very preterm neonates with respiratory distress syndrome.

BACKGROUND: Vasomotor nephropathy is a common renal dysfunction in very preterm neonates. OBJECTIVE: To determine whether theophylline could prevent vasomotor nephropathy in very preterm infants with respiratory distress syndrome. METHODS: A randomised, double blind, placebo controlled trial of 50 preterm infants of gestational age < or = 32 weeks needing assisted ventilation. Infants received an intravenous dose of theophylline (1 mg/kg) or placebo for three days. The 24 hour urine volume was measured daily. On days 2, 5, and 11, blood samples and 12 hour urine collections were analysed for electrolytes, creatinine, and urea. RESULTS: On day 1, urine output was significantly higher in the theophylline (2.4 (0.9) ml/kg/h) than the placebo (1.6 (1.0) ml/kg/h; p = 0.023) group (values are mean (SD)). The incidence of oligoanuria was significantly lower in the theophylline treated (5%) than the placebo (33%) group. Twenty four hours after the first administration of theophylline/placebo, serum creatinine concentration was significantly lower in the theophylline (0.76 (0.23) mg/dl) than the placebo (1.0 (0.41) mg/dl; p = 0.025) group. On day 5 an increase in serum creatinine was observed in both groups. On day 11 a significant reduction in serum creatinine was observed, compared with day 5, with no difference between the two groups. CONCLUSION: The results suggest that, in very preterm infants with respiratory distress syndrome, early theophylline administration improves renal function during the first two days of life.

Effect of passive smoking on thyroid function in infants.

OBJECTIVE: To evaluate the effect of passive smoking on thyroid function in infants. DESIGN AND METHODS: Cord serum tri-iodiothyronine (T3), free T3 (fT3), thyroxine (T4), free T4 (fT4), TSH, thyroxine binding globulin (TBG), thyroglobulin (TG) and cord plasma thiocyanate were measured at birth, and serum TG and thiocyanate after 1 year of life, in 18 infants born from parents who did not smoke (group A), in 18 infants with a father who smoked (group B) and in 18 infants with parents both being smokers (group C). RESULTS: No significant differences were observed in cord serum concentrations of T3, fT3, T4, fT4, TSH and TBG among the three groups. Median (range) TG concentrations (ng/ml) were 30.2 (5.0-102.0), 56.3 (20.5-208.0) and 76.0 (26.0-199.0) at birth (P=0.009 for groups A and B compared; P=0.0002 for groups A and C compared), and 14.9 (5.4-32.0), 19.5 (10.0-57.5) and 20.0 (14.0-40.7) at 1 year (P=0.017 for groups A and C compared), in the three groups respectively, and thiocyanate concentrations (mmol/l) were 3.3 (0.0-51.4), 12.9 (0.0-122.2) and 27.8 (3.3-184.5) at birth (P=0.015 for groups A and C compared), and 3.1 (0.0-32.7), 6.0 (0.0-47.3) and 20.3 (0.0-230.8) at 1 year (P=0.01 for groups A and C compared) in the three groups respectively. CONCLUSIONS: TG and thiocyanate concentrations at birth and at 1 year of age in infants of smoking parents are greater than in infants with non-smoking parents. These results indicate that the change in thyroid function as evaluated by serum TG concentrations observed at birth can persist at least for 1 year if the exposure to passive smoking from both parents is continued. Increased TG concentrations may be due to a direct effect of thiocyanate on the thyroid gland.

Single molecule studies by means of the two-photon fluorescence distribution.

We have characterized a commercial confocal scanning head for the detection of single molecule fluorescence by two-photon excitation. We have verified that the distribution of the fluorescence emitted by dyes and labeled proteins on glass substrates is discrete with quanta proportional to a common reference signal. We describe and test a simple and quantitative tool to discriminate between single molecules and molecular aggregates on single snapshots based on the analysis of the intensity distribution. We have verified the square dependence of the fluorescence intensity vs. the excitation power, suggesting that no appreciable saturation and fast photo-damage of the chromophores takes place at the excitation power employed here.

Two-photon microscopy and spectroscopy based on a compact confocal scanning head.

We have combined a confocal laser scanning head modified for TPE (two-photon excitation) microscopy with some spectroscopic modules to study single molecules and molecular aggregates. The behavior of the TPE microscope unit has been characterized by means of point spread function measurements and of the demonstration of its micropatterning abilities. One-photon and two-photon mode can be simply accomplished by switching from a mono-mode optical fiber (one-photon) coupled to conventional laser sources to an optical module that allows IR laser beam (two-photon/TPE) delivery to the confocal laser scanning head. We have then described the characterization of the two-photon microscope for spectroscopic applications: fluorescence correlation, lifetime and fluorescence polarization anisotropy measurements. We describe the measurement of the response of the two-photon microscope to the light polarization and discuss fluorescence polarization anisotropy measurements on Rhodamine 6G as a function of the viscosity and on a globular protein, the Beta-lactoglobulin B labeled with Alexa 532 at very high dilutions. The average rotational and translational diffusion coefficients measured with fluorescence polarization anisotropy and fluorescence correlation methods are in good agreement with the protein size, therefore validating the use of the microscope for two-photon spectroscopy on biomolecules.

Influence of ligands on the fluorescence polarisation anisotropy of ethidium bound to DNA.

The decay of the fluorescence polarisation anisotropy (FPA) of the ethidium-DNA complex has been measured by multifrequency phase fluorometry, in order to study the perturbations induced by the presence of different ligands on the torsional dynamics of DNA, a moderately flexible polymer that undergoes bending (flexure of the helix axis) and torsional (twisting of base pairs) motions. Two probes have been used together with ethidium: an intercalator, chloroquine, and a minor groove binding dye: hoechst 33258. Chloroquine is found to substantially modify the DNA torsional dynamics both in linear and in circularly closed DNAs only at high binding ratios, in agreement with previous reports [Wu et al. Biochem. 27 (1988) 8128]. The effective elastic constant becomes approximately three times larger when the dye/base pairs binding ratio is higher than 0.14. The minor groove ligand hoechst 33258, on the other hand, greatly increases the effective elastic constant to the point that at dye/base pairs ratios larger than 0.5, the effective elastic constant becomes stiffer by several orders of magnitude, suggesting a progressive hindering of internal motions. The results reported here show that DNA torsions are more effectively influenced by groove-binding molecules than by intercalators and it is expected that the large perturbation of the former ligand may be useful when describing the change in the dynamical properties induced by DNA binding proteins. FPA in the frequency domain, the technique adopted throughout this work, has proved to be very sensitive to changes of the elastic constant that describes DNA torsional dynamics. Several computer simulations performed in order to predict the FPA time decay of intercalated ethidium have led to good agreement with the observed results.

Conformation of intercalated DNA plasmids investigated by circular dichroism and dynamic light scattering.

Two DNA plasmids, pEGF and pACL29, intercalated with ethidium bromide (EB), have been examined by circular dichroism (CD) and dynamic light scattering (DLS). CD and DLS data show significant changes when the EB/DNA (phosphates) ratio reaches a value of r approximately equal to 0.13. The translational and rotational diffusion coefficients, predicted assuming that plasmids can be described by a string of beads, and the CD spectrum, suggest that a transition from an interwound to a toroidal conformation is likely to occur.

Primitive sinonasal malignant mucosal melanoma: description of a case treated with radiotherapy (0-7-21 regimen).

Mucosal malignant melanoma (MMM) of the nasal cavity and paranasal sinuses is rare and has a poor prognosis. The rarity of MMM of the ethmoid is also noteworthy and primary treatment has been, till now, surgery and/or radiotherapy (RT). Here we report a case of MMM arising in the ethmoid of a caucasian man and treated with hypofractionated RT. A 32 year-old man presented with symptoms of cephalalgia, obstruction and nasal bleeding; a computed tomography (CT) showed a large mass that involved ethmoid, left orbit and roof of the nasal cavity. After biopsy, a MMM was found. Chest radiography demonstrated the presence of multiple lung metastases and still the patient was submitted to palliative radiotherapy according to 0-7-21 regimen with a total dose of 24Gy/3 fraction/21dd, dose per fraction 8Gy. The patient was asymptomatic one month after the end of RT and three months later a CT demonstrated a partial remission. The patient died 17 months after the initiation of RT for disseminated disease, without clinical signs of tumoral regrowth in the irradiated site. This case confirms the efficacy and the safety of 0-7-21 RT regimen; the absence of symptoms after 17 months and the poor prognosis encourage the use of RT as primary treatment for MMM of the head and neck.

Autoimmune haemolytic anaemia in a newborn infant.

The case is reported of an infant with autoimmune haemolytic anaemia of perinatal onset. Combined treatment with steroids and cyclosporin was necessary to improve haemolysis and reduce the high transfusion requirements. Treatment was discontinued at 13 months of age. The child was healthy at the follow up at 24 and 36 months of age.

Increased renal echogenicity in the neonate.

We report data on newborn infants with increased renal echogenicity observed at the Division of Neonatal Intensive Care of Pavia during a five-year period. Review of 1600 abdominal ultrasonic evaluations revealed 103 newborn infants (56 females and 47 males, with birth weight from 560 to 3700 g and gestational age from 25 to 42 weeks) whose kidneys showed increased echogenicity. Three patients with infantile polycystic kidney disease, two with renal candidiasis, three with dysplastic kidney and two with renal vein thrombosis showed diffuse hyperechogenicity. Three patients with hemolytic-uremic syndrome showed cortical hyperechogenicity. Increased medullary echogenicity was observed in 90 infants with renal disease secondary to perinatal asphyxia. In 76 of these patients the evaluation of renal echogenicity and the renal function improved, while in the remaining 14 newborns the renal alteration persisted until death.

MR findings in thyroglossal duct cysts: report of two cases.

Two patients with thyroglossal duct cysts have been studied with CT and MR. The typical CT feature of these cystic upper-neck lesions are depicted in literature, conversely MR findings are not well known. The homogeneous high intensity on T1-weighted images, higher than simple cyst or fluid, is the most typical feature of the thyroglossal cyst.