In vitro pharmacological characterization of standard and new lysophosphatidic acid receptor antagonists using dynamic mass redistribution assay.

Lysophosphatidic acid (LPA) is a bioactive phospholipid that acts as an agonist of six G protein-coupled receptors named LPA receptors (LPA1-6). LPA elicits diverse intracellular events and modulates several biological functions, including cell proliferation, migration, and invasion. Overactivation of the LPA-LPA receptor system is reported to be involved in several pathologies, including cancer, neuropathic pain, fibrotic diseases, atherosclerosis, and type 2 diabetes. Thus, LPA receptor modulators may be clinically relevant in numerous diseases, making the identification and pharmacodynamic characterization of new LPA receptor ligands of strong interest. In the present work, label-free dynamic mass redistribution (DMR) assay has been used to evaluate the pharmacological activity of some LPA1 and LPA2 standard antagonists at the recombinant human LPA1 and LPA2 receptors. These results are compared to those obtained in parallel experiments with the calcium mobilization assay. Additionally, the same experimental protocol has been used for the pharmacological characterization of the new compound CHI. KI 16425, RO 6842262, and BMS-986020 behaved as LPA1 inverse agonists in DMR experiments and as LPA1 antagonists in calcium mobilization assays. Amgen compound 35 behaved as an LPA2 antagonist, while Merck compound 20 from WO2012028243 was detected as an LPA2 inverse agonist using the DMR test. Of note, for all the compounds, similar potency values were estimated by DMR and calcium assay. The new compound CHI was found to be an LPA1 inverse agonist, but with potency lower than that of the standard compounds. In conclusion, we have demonstrated that DMR assay can be successfully used to characterize LPA1 and LPA2 ligands. Compared to the classical calcium mobilization assay, DMR offers some advantages, in particular allowing the identification of inverse agonists. Finally, in the frame of this study, a new LPA1 inverse agonist has been identified.

Response of central serous chorioretinopathy evaluated by multimodal retinal imaging.

PurposeTo identify predictive biomarkers of treatment outcomes by multimodal retinal imaging in patients affected by central serous chorioretinopathy (CSC).Patients and methodsIn this interventional non-randomized clinical study, 27 treatment-naive CSC patients were prospectively enrolled and treated with oral eplerenone for 5-13 weeks. Primary outcomes included presence of pathological findings on indocyaine green angiography (ICGA), structural optical coherence tomography (OCT) and OCT-angiography (OCT-A) at baseline associated with different response to the treatment.ResultsA total of 29 eyes of 27 patients (2 females, 25 males) met the inclusion criteria and were included in the study (mean age was 45±7 years). Mean CSC duration at baseline was 13.5±4.4 weeks. After a mean of 10.5 weeks of treatment, mean central macular thickness significantly reduced (P<0.001), and mean best-corrected visual acuity improved (P<0.001). Seventeen eyes (61%) demonstrated total reabsorption of subretinal fluid on structural OCT, five eyes (18%) presented a partial response to eplerenone therapy and six eyes (21%) showed no response. The complete response to the treatment was associated with absence of CNV at OCT-A and the presence of hotspot at ICGA (P<0.001 and P=0.002, respectively). None of eight eyes with CNV in OCT-A imaging had a complete response to eplerenone and none of three eyes without hotspot at ICGA showed a complete response to the treatment.ConclusionsMultimodal retinal imaging allowed us to propose predictive biomarkers (ie, absence of CNV on OCT-A and presence of hotspot on ICGA) for treatment outcomes.

[Hospital readmission after postpartum discharge of term newborns in two maternity wards in Stockholm and Marseille]. / Morbidité néonatale précoce après sortie de maternité : étude comparative entre deux maternités à Stockholm et Marseille.

The consequences of early postpartum discharge (EPPD, within 2 days after birth) on newborn health remain debated. Early discharge has been associated with increased neonatal morbidity. However, neonatal re-hospitalization can be prevented by careful follow-up during the 1st week after birth. We compared the early neonatal hospitalization of term newborns over 2 years in two hospitals: Karolinska University Hospital in Stockholm (n=7300 births), which allowed early discharge from 6h after birth with specific neonatal follow-up, and Marseille University Hospital (AP-HM) (n=4385) where postpartum discharge was more conventional after 72 h. During the study period, the EPPD rate was 41% vs. 2% in Stockholm and Marseille, respectively (P<0.001). Hospital readmission was comparable (5.6‰ vs. 7‰, P=0.2). The leading cause associated with hospitalization was icterus in Stockholm (76% vs. 26%, P<0.001) and feeding difficulties in Marseille (17% vs. 48%, P<0.001). In conclusion, close neonatal follow-up during the 1st week of life associated with restricted maternal and neonatal eligibility criteria for EPPD are required to prevent early neonatal re-hospitalization.

Corneal transplantation activity over 7 years: changing trends for indications, patient demographics and surgical techniques from the Corneal Transplant Epidemiological Study (CORTES).

This study aims to examine evolving indications and changing trends for corneal transplantation in Italy. Corneal transplantations performed with donor tissues distributed by the Veneto Eye Bank Foundation between 2002 and 2008 were prospectively evaluated. Of the 13,173 keratoplasties performed on 11,337 patients, 10,742 (81.5%) were penetrating (PK), 1644 (12.5%) were anterior lamellar (ALK), and 787 (6.0%) were endothelial (EK). Keratoconus (42.5%), regraft (18.9%), and pseudophakic bullous keratopathy (PBK, 11.9%) were the leading indications for PK, with keratoconus (69.6%) and regraft (6.5%) showing higher indications for ALK, whereas pseudophakic bullous keratopathy (50.1%) and regraft (18.7%) were the major indications for EK. There was an overall decrease observed in corneal grafting for keratoconus (P = .0048) and an increase for PBK (P = .0653) and regrafting (P = .0137). These indications differed by age and gender. The number of keratoplasties over 7 years was stable (P = .2394), although the annual number of PKs declined by 34.0% (P = .0250), ALKs began to rise from 2005 (P = .0600), whereas EKs showed a huge growth, with their number tripling in 2007 and further doubling in 2008 (P = .0004). Leading indications for keratoplasty showed similar data that have been reported elsewhere for Western countries over the past few decades, albeit with a higher percentage of keratoconus. However, the overall number of keratoplasties for keratoconus was in decline, whereas regraft keratopathy and PKs increased due to the application of the newer surgical techniques for corneal grafting. This highlights an important shift in managing corneal diseases toward the application of selective and more conservative surgeries and changes in indications in corneal transplantation.

Discovery of a novel isoxazoline derivative of prednisolone endowed with a robust anti-inflammatory profile and suitable for topical pulmonary administration.

A novel glucocorticoids series of (GCs), 6α,9α-di-Fluoro 3-substituted C-16,17-isoxazolines was designed, synthesised and their structure-activity relationship was evaluated with glucocorticoid receptor (GR) binding studies together with GR nuclear translocation cell-based assays. This strategy, coupled with in silico modelling analysis, allowed for the identification of Cpd #15, an isoxazoline showing a sub-nanomolar inhibitory potency (IC50=0.84 nM) against TNFα-evoked IL-8 release in primary human airways smooth muscle cells. In Raw264.7 mouse macrophages, Cpd #15 inhibited LPS-induced NO release with a potency (IC50=6 nM)>10-fold higher with respect to Dexamethasone. Upon intratracheal (i.t.) administration, Cpd #15, at 0.1 µmol/kg significantly inhibited and at 1 µmol/kg fully counteracted eosinophilic infiltration in a model of allergen-induced pulmonary inflammation in rats. Moreover, Cpd #15 proved to be suitable for pulmonary topical administration given its sustained lung retention (t1/2=6.5h) and high pulmonary levels (>100-fold higher than plasma levels) upon intratracheal administration in rats. In summary, Cpd #15 displays a pharmacokinetic and pharmacodynamic profile suitable for topical treatment of conditions associated with pulmonary inflammation such as asthma and COPD.

A case of Beauveria bassiana keratitis confirmed by internal transcribed spacer and LSU rDNA D1-D2 sequencing.

We describe a case of fungal keratitis due to Beauveria bassiana in a farmer with Fuchs' dystrophy, treated with amphotericin B. Surgery with penetrating keratoplasty was necessary to resolve the lesions. Susceptibility testing and molecular sequencing permitted the identification and treatment of this rare aetiological agent of invasive fungal disease.

[Nuchal translucency and combined test: what are the implications in clinical practice?]. / La translucenza nucale e il test combinato: quali risvolti nella pratica clinica?

AIM: The aim of this study was to evaluate the performance of the combined test (nuchal translucency, NT) and maternal serum free-beta human chorionic gonadotropin (free beta-hCG) and pregnancy-associated plasma protein-A (PAPP-A), compared to the NT measurement alone, in fetal aneuploidy screening in the general population and in pregnant women aged 35 years and over. In addition, the association between increased NT and presence of cardiac defects in fetuses with normal karyotype was evaluated. METHODS: Screening at 11-14 weeks of gestation by NT measurement and combined test was carried out in 1521 pregnant women. The estimated risk for trisomy 21 and trisomy 13+18 was calculated (risk cut-off 1/300 and 1/750 respectively) and the outcomes was evaluated. RESULTS: Ten cases of trisomies (21 and 18) occurred, seven of which among the older group of pregnant women. The detection rate (DR) for the combined test was 80% in the general population and 85.7% in older pregnant women, which resulted higher rate than NT measurements alone. Detection rate of cardiac defects using NT measurements was 66.6%. CONCLUSION: The combined test is an effective screening for aneuploidies and reduces at 14% the need of invasive testing in the older obstetric population, detecting all the trisomies occurred in this group. The association between increased NT and cardiac defects is confirmed but it seems too weak to consider NT as a single screening strategy for these abnormalities.

Maternal plasma level of antimicrobial peptide LL37 is a major determinant factor of neonatal plasma LL37 level.

AIM: To determine cathelicidin antimicrobial peptide LL37subcellular distribution in cord neutrophils and normal plasma LL37 levels in mothers and neonates, relate them to delivery mode and relevant biochemical markers, including 25-OHvitamin D [25(OH)D] as this molecules increases cathelicidin gene expression. METHODS: A total of 115 infants were included, n = 68 with normal delivery and n = 47 with elective Caesarean section (C-section), a subset of these being 50 mother-infant pairs. Biomarkers were determined in maternal and cord blood. Subcellular peptide LL37 distribution was analysed with immunoelectron microscopy. RESULTS: Cord plasma LL37 levels were three-times higher after normal delivery compared with C-section. A highly significant correlation was observed between maternal and cord plasma LL37 levels, regardless of delivery mode. No relationship was found between LL37 and 25(OH)D levels. Neutrophils from cord blood after normal delivery contained 10-times more cytoplasmatic cathelicidin peptide compared with corresponding cells after C-section where a strict granular localization was found. CONCLUSION: These data are consistent with a placental transfer of LL37 and identifies maternal stores as the critical factor determining neonatal plasma LL37 level. An additional enhancement of neonatal cathelicidin mobilization and release is connected to normal delivery stress.

Human cathelicidin peptide LL37 inhibits both attachment capability and biofilm formation of Staphylococcus epidermidis.

AIMS: The aim of this work was to investigate the possible effect of human cathelicidin antimicrobial peptide LL37 on biofilm formation of Staphylococcus epidermidis, a major causative agent of indwelling device-related infections. METHODS AND RESULTS: We performed initial attachment assay and biofilm formation solid surface assay in microtitre plates, as well as growth experiment in liquid medium using laboratory strain Staph. epidermidis ATCC35984. We found that already a low concentration of the peptide LL37 (1 mg l(-1)) significantly decreased both the attachment of bacteria to the surface and also the biofilm mass. No growth inhibition was observed even at 16 mg l(-1) concentration of LL37, indicating a direct effect of the peptide on biofilm production. CONCLUSIONS: As biofilm protects bacteria during infections in humans and allows their survival in a hostile environment, inhibition of biofilm formation by LL37 may have a key role to prevent bacterial colonization on indwelling devices. SIGNIFICANCE AND IMPACT OF THE STUDY: Our findings suggest that this host defence factor can be a potential candidate in prevention and treatment strategies of Staph. epidermidis infections in humans.

Gas tamponade and cyclocryotherapy of a chronic cyclodialysis cleft.

AIMS: To describe a minimally invasive technique to treat a chronic large cyclodialysis cleft that had failed to respond to medical therapy. METHODS: A 51-year-old man with a history of blunt trauma developed a unilateral chronic ocular hypotony. He was treated with topical atropine 1% for 3 months. 12 months later, the patient was referred to our glaucoma service for evaluation and treatment of persistent hypotony. Ultrasound biomicroscopy (UBM) displayed a cyclodialysis cleft extending from the 6 to 12 o'clock positions. B-scan echography revealed a peripheral choroidal effusion. A single bubble of 20% sulfur hexafluoride was injected into the vitreous cavity and transconjunctival cyclocryotherapy was performed. RESULTS: After gas absorption, intraocular pressure increased to 12 mm Hg and became steady during the follow-up. B-scan echography showed the disappearance of choroidal effusion, and UBM displayed a complete closure of the cyclodialysis cleft from the 6 to 8:30 o'clock positions and from the 9:30 to 12 o'clock positions. A small cleft extending from the 8:30 to the 9:30 positions remained after the treatment, but the distance between the scleral spur and the ciliary boby decreased, and the cleft was limited at the back due to the scar formation. CONCLUSION: Gas tamponade with cyclocryotherapy represents a minimally invasive technique that is worth considering for patients with cyclodialysis clefts associated with a shallow anterior chamber and that had failed to respond to medical therapy. This technique should be useful in cases of cyclodialysis clefts that are not amenable to treatment with more conservative efforts.

Comparison of Goldmann applanation tonometry and dynamic contour tonometry in healthy and glaucomatous eyes.

PURPOSE: To compare the measurements of intraocular pressure (IOP) with dynamic contour tonometry (DCT) and Goldmann applanation tonometry (GAT) in healthy and glaucomatous eyes and to evaluate the influence of corneal thickness (CCT), corneal curvature (CC), axial length (AL), and age on these tonometric alternatives. METHODS: Three hundred eyes of 100 healthy subjects, 100 patients with primary open angle glaucoma, and 100 patients with primary angle-closure glaucoma underwent IOP evaluation with DCT and GAT, and measurements of CCT, CC, and AL. Bland-Altman plots were used to evaluate the agreement between tonometers. Regression analysis was used to evaluate the influence of ocular structural factors on IOP measurements obtained with both tonometers. RESULTS: Bland-Altman plots indicated that the 95% limits of agreement between tonometers were -1.4 to 6.6 mmHg. DCT values measured 2.6+/-1.9 mmHg higher than GAT readings (P<0.001). The mean IOP difference between DCT and GAT (DeltaIOP) was higher in healthy than in glaucomatous eyes and the magnitude of difference increased with increasing CCT. A significant reduction of DeltaIOP with an increase in both CCT (P<0.001) and IOP values (P<0.001) was found. Regression analysis showed no effect of CC, AL, and age on both DCT and GAT readings. In contrast to GAT (P<0.001), DCT measurements were not influenced by CCT (P=0.43). CONCLUSIONS: IOP readings obtained by DCT were higher and less affected by CCT than those by GAT. The DeltaIOP was higher in healthy than in glaucomatous eyes and decreased in relation to increased CCT and IOP values.

The effect of pneumatic trabeculoplasty on intraocular pressure: the results of a 6-month, open-label, multicenter study.

PURPOSE: To evaluate the effects of pneumatic trabeculoplasty (PNT) in ocular hypertension and glaucoma subjects. METHODS: A total of 63 consecutive subjects, either treated (79%) or untreated (21%), with intraocular pressure (IOP) between 20 and 25 mmHg were enrolled; the eye with higher IOP (or, in case of identical IOP, worse visual field) was treated with PNT, with the fellow eye used as control. Subjects underwent a baseline evaluation the day before treatment, two PNT treatments at day 0 and 7, visits at day 1, 8, 14, and at each month until the end of the study, which lasted 6 months. Safety was addressed at all visits; an IOP curve (at 8 and 10 AM, 2 and 4 PM) was obtained at baseline and during monthly visits. RESULTS: In PNT eyes, baseline IOP was 22.2-/+1.6 mmHg. Following PNT a statistically significant reduction of IOP occurred at all visits (p<0.0001), with a mean decrease ranging from -2.7-/+2.5 (-11.9-/+10.8%) to -3.6-/+2.6 mmHg (-16.0-/+11.6%); mean reduction was 12.8-/+11.5%. Although IOP diminished also in the control eyes after baseline (p<0.05), the change in IOP was significantly higher in PNT group at each visit (p<0.05). Mild side effects were experienced by 76% of subjects and they all resolved without sequelae. CONCLUSIONS: The results suggest the effect of this procedure in reducing IOP in glaucoma and ocular hypertensive subjects.

Simultaneous bilateral retinal detachment following coronary artery bypass graft: case report.

PURPOSE: To present an unusual case of simultaneous bilateral retinal detachment (RD) following a coronary artery bypass graft in a patient with acute myocardial infarction (AMI). METHODS: A 78-year-old man was first seen for bilateral sudden visual loss after surgical treatment of AMI. The patient underwent ultrasound biomicroscopy (UBM) and ocular B-scan echographic examination. RESULTS: The ocular assessment showed a bilateral seclusion of the pupil with bombe of the iris, an anterior chamber without cells or flare, and hypotonia. The evaluation of the visual acuity revealed no light perception in the right eye (RE) and uncertain light perception in the left eye (LE). The UBM analysis of the anterior segment confirmed the presence of bilateral pupillary block due to the seclusion of the pupil and a peripheral serous choroidal detachment involving the RE. The echographic B-scan analysis of the posterior segment showed a bilateral closed funnel-shaped RD and confirmed the presence of the peripheral flat serous choroidal detachment in RE. CONCLUSIONS: The cause for simultaneous bilateral RD remained unclear. It may have been a consequence of a persistent choroidal detachment with multiple swelling and 'kissing' of retinal surface. The increased venous pressure caused by congestive heart failure due to AMI could have caused a bilateral uveal effusion. Alternatively, the absence of retinal tears, the presence of a closed funnel-shaped morphology, and seclusion of the pupils allowed us to suspect an exudative pathogenetic mechanism due to a previous unrecognized ocular inflammatory state.

An unusual case of pneumoperitoneum: nephrocolic fistula due to a giant renal staghorn calculus.

Gastrointestinal perforations usually lead to pneumoperitoneum and peritonitis. Rarely, if ever described, a complete giant staghorn renal stone might cause a nephrocolic fistula with sigmoid impaction and perforation similar to gallstone ileus. Few nephrointestinal fistulae have been described in the literature and none of them were presented as an acute abdomen with pneumoperitoneum and pneumoretroperitoneum. To our knowledge, this is the only case showing CT and radiographic findings of a pathology not yet described in the literature. We named the sigmoid perforation by a renal stone ileus "Lorenzi's syndrome" after the physician who hypothesized this rare differential diagnosis based only on history and clinical examination.

Efficacy of the symmetric, patient-oriented, rigid, three-dimensional, active (SPoRT) concept of bracing for scoliosis: a prospective study of the Sforzesco versus Lyon brace.

AIM: Bracing is considered to be effective in the treatment of adolescent idiopathic scoliosis. The concept prevailing today includes an asymmetrical construction, mainly using a mechanical three-point system. We developed the new Sforzesco brace, based on the SPoRT concept (Symmetric, Patient-oriented, Rigid, Three-dimensional, active). The aim of this study is to verify the results of this concept and brace, compared to three-point classical systems. METHODS: We performed a prospective, pair-controlled study. It was possible to match 15 patients, out of the first 18 consecutively treated with the Sforzesco brace (Group SPoRT), with previous patients treated with the Lyon brace (Group LY). SPoRT included 14.2+/-1.7 year old patients, with 47+/-7 degrees Cobb worst curves, and 11+/-4 degrees Bunnell; LY included 13.6+/-1.6 year old patients, with 43+/-7 degrees Cobb and 12+/-5 degrees Bunnell. The brace had to be worn 23 h per day and patients were evaluated after 6 months both clinically and radiographically (without the brace). Appropriate statistics were used. RESULTS: At the baseline there were slight differences between the groups, with SPoRT worse than LY. All radiographic and clinical parameters decreased significantly with treatment in both groups, apart from thoracic Cobb degrees in LY. SPoRT had better results than LY (P<0.05) radiographically (worst curve -10+/-5 degrees vs -5+/-7 degrees, all curves -8+/-7 degrees vs -6+/-7 degrees), for sagittal profile (distance from plumbline: T12 -6+/-9 mm vs +2+/-8 and L3 -7+/-12 vs 0+/-10) and aesthetics of the shoulders (9 improved and 6 unchanged vs 5 and 8) and waists (10 improved and 5 unchanged vs 5 and 8). Finally, clinical results in terms of patient recovery were better in SPoRT than LY (12 improved and 3 unchanged vs 8 and 5). CONCLUSION: When a new treatment is introduced, it is not possible to wait years (end of therapy) before verifying its utility, and in scoliosis bracing a short term study already gives very important clues. This study confirms the immediate efficacy of brace treatment (even in such high degree cases) whatever the brace concept used, with only 2 (out of 30) progressed curves. In SPoRT we had no progressions, and obtained a statistically significant 80% better radiographic results than LY in the worst scoliosis curve, and 40% in the average of all curves, as well as improved aesthetics and sagittal profile (that is crucial in scoliosis bracing). The Sforzesco brace should be used, and the SPoRT concept explored in the long term to verify if the classical three-point system should be totally (or partially) abandoned.

Influence of pancreas transplantation alone on native renal function.

Pancreas transplantation alone (PTA) has become an accepted treatment of nonuremic diabetic patients, when the risks of secondary complications of diabetes mellitus are greater than those of the surgical procedure and the posttransplant immunosuppression. As a decrease in native renal function is expected, we followed this parameter among patients who underwent PTA. From January 1997 through January 2005, we performed 69 PTA in 66 patients. All patients showed glucose hyperlability with hypoglycemic unawareness, or two or more diabetic complications as well as creatinine clearance (CrCl) > or = 45 mL/min. Immunosuppression was based on tacrolimus, mycophenolate mofetil and prednisone. Twenty-four hour CrCl were performed after all successful PTA. We divided patients in two groups according to the pretransplant CrCl: group 1, CrCl < or = 70 mL/min (n = 20) and group 2, CrCl > 70 mL/min (n = 25). The data were analyzed using Student's t-test (P < or = .05 was considered significant). Twenty-one patients were excluded from the analysis because of death (n = 5) or graft loss (n = 8) during the first year or follow-up shorter than 1 year (n = 8). The mean value of CrCl decreased 28.8% (85.0 +/- 31 versus 60.5 +/- 36 mL/min; P < .001). There was also a 39.3% reduction among group 1 subjects (P = .003), including 10 who displayed CrCl < or = 30 mL/min. There was also a 24.4% reduction among group 2 (P = .008), but no patient developed end-stage renal disease. In conclusion, native renal function decreased significantly after PTA, but was well tolerated among patients with CrCl > 70 mL/min. Patients with CrCl < 70 mL/min show a significant risk of worsened renal function.

Enteric conversion after bladder drained pancreas transplantation experience of 14 cases.

The method of exocrine diversion in pancreas allograft continues to be controversial due to the advantages versus disadvantages of bladder versus enteric techniques. Bladder drainage (BD) exposes the patient to urological and metabolic problems that may require conversion to enteric drainage (ED). The purpose of this study was to review our initial experience of conversion from BD to ED for patients who underwent pancreas transplantation originally with bladder diversion. Among 114 pancreas transplantation performed with BD, from January 1996 to April 2003, 60 were simultaneous pancreas-kidney transplantation (SPKT), 35 were pancreas transplantation alone (PA), and 19 were pancreas after kidney transplantations (PAK). Twenty-three (20.2%) cases were excluded due to early death of the patient or the graft, yielding an analyses of 91 patients. Enteric conversion (EC) was performed in 14 (15.4%) patients with a mean follow-up of 15.7 months (range, 3-51 months) after transplantation including 8 (8.8%) SPKT, 4 (4.4%) PAK, and 2 (2.2%) PA. No surgical morbidity or mortality was observed related to EC. All patients had complete resolution of the initial problem with preservation of pancreatic function. EC represents an easy, safe procedure with low morbidity and mortality rates, representing the option of choice for patients with persistent urological or metabolic disturbances.

The newborn infant is protected by an innate antimicrobial barrier: peptide antibiotics are present in the skin and vernix caseosa.

BACKGROUND: Peptide antibiotics are part of the surface defences against microbial intruders. However, the presence and significance of these innate immune effectors in the skin barrier of the newborn infant have not yet been appreciated. Erythema toxicum neonatorum is an inflammatory skin reaction of unknown aetiology and significance, commonly present in the healthy newborn infant. OBJECTIVES: As peptide antibiotics are upregulated in inflammatory skin disorders, we hypothesized that this also could be the case in erythema toxicum. We also investigated if the vernix caseosa, a cream-like white substance present on the skin of the infant at birth, might contribute to host defences. METHODS: The presence of the human antibacterial peptide LL-37 was investigated by immunohistochemistry and confocal imaging of skin biopsies from four 1-day-old infants with an erythema toxicum rash and four matched newborns without the rash. In addition, we analysed the expression of LL-37 and human beta defensin-1, an antibacterial peptide of epithelial origin, by reverse transcriptase-polymerase chain reaction. Finally, we screened for antibacterial components in vernix material obtained from six healthy newborns by inhibition zone assays. RESULTS: All biopsies from the lesions of erythema toxicum showed a dense, nodular infiltrate with numerous LL-37-expressing cells located in the dermal layer and a clear localization of the peptide within CD15-expressing neutrophils, EG2-expressing eosinophils and CD1a-expressing dendritic cells. LL-37 was also found to be located in CD1a-expressing Langerhans cells and a positive staining for the peptide was seen throughout the whole epidermal layer, both in infants with and without the rash. Skin samples from infants with the rash of erythema toxicum showed a constitutive expression of human beta defensin-1, while the expression of LL-37 seemed to be induced. Furthermore, LL-37 and lysozyme were detected in the protein fractions derived from the vernix caseosa, and these fractions exhibited a clear antibacterial activity. CONCLUSIONS: Peptide antibiotics are present in the vernix caseosa and in the skin of the healthy newborn infant, indicating effective innate immune protection already during fetal and neonatal life.