Effects of Topical Application of CHF6467, a Mutated Form of Human Nerve Growth Factor, on Skin Wound Healing in Diabetic Mice.

Nerve growth factor (NGF) is the protein responsible for the development and maintenance of sensory skin innervation. Given the role of appropriate innervation in skin healing, NGF has been indicated as a possible prohealing treatment in pathologic conditions characterized by nerve-ending loss, such as chronic ulcers in diabetes; however, its use as a therapeutic agent is limited by its hyperalgesic effect. We tested the effect of topical application of the nonalgogenic NGF derivative hNGFP61S/R100E in two models of skin ulcer induced in dbdb diabetic mice, investigating healing time, skin histology, reinnervation, and angiogenesis using morphologic and molecular approaches. We showed that the topical administration of CHF6467, a recombinant human NGF in which an amino acid substitution (R100E) abolished the hyperalgesic effect usually associated with NGF, accelerated skin repair in experimental wounds (full-excision and pressure-ulcer) induced in diabetic mice (dbdb). CHF6467-induced acceleration of wound healing was accompanied by increased re-epithelization, reinnervation, and revascularization as assessed by histology, immunohistochemistry, and image analysis. Bioinformatic analysis of differentially expressed genes and signaling pathways in the wound tissues showed that protein kinase B-mammalian target of rapamycin was the most regulated pathway. In spite of the transdermal absorption leading to measurable, dose-dependent increases in CHF6467 plasma levels, no systemic thermal or local mechanical hyperalgesia was observed in treated mice. When tested in vitro in human cell lines, CHF6467 stimulated keratinocyte and fibroblast proliferation and tube formation by endothelial cells. Collectively, these results support a possible use of CHF6467 as a prohealing agent in skin lesions in diabetes. SIGNIFICANCE STATEMENT: Topical application of CHF6467 accelerates reinnervation, neoangiogenesis, and wound healing in diabetic mice in both full-thickness skin-excision and pressure-ulcer models through the protein kinase B/mammalian target of rapamycin pathway and does not induce hyperalgesia.

A ring system detected around the Centaur (10199) Chariklo.

Hitherto, rings have been found exclusively around the four giant planets in the Solar System. Rings are natural laboratories in which to study dynamical processes analogous to those that take place during the formation of planetary systems and galaxies. Their presence also tells us about the origin and evolution of the body they encircle. Here we report observations of a multichord stellar occultation that revealed the presence of a ring system around (10199) Chariklo, which is a Centaur--that is, one of a class of small objects orbiting primarily between Jupiter and Neptune--with an equivalent radius of 124 ± 9 kilometres (ref. 2). There are two dense rings, with respective widths of about 7 and 3 kilometres, optical depths of 0.4 and 0.06, and orbital radii of 391 and 405 kilometres. The present orientation of the ring is consistent with an edge-on geometry in 2008, which provides a simple explanation for the dimming of the Chariklo system between 1997 and 2008, and for the gradual disappearance of ice and other absorption features in its spectrum over the same period. This implies that the rings are partly composed of water ice. They may be the remnants of a debris disk, possibly confined by embedded, kilometre-sized satellites.

From bench to bedside: in vitro and in vivo evaluation of a neonate-focused nebulized surfactant delivery strategy.

BACKGROUND: Non-invasive delivery of nebulized surfactant has been a neonatology long-pursued goal. Nevertheless, the clinical efficacy of nebulized surfactant remains inconclusive, in part, due to the great technical challenges of depositing nebulized drugs in the lungs of preterm infants. The aim of this study was to investigate the feasibility of delivering nebulized surfactant (poractant alfa) in vitro and in vivo with an adapted, neonate-tailored aerosol delivery strategy. METHODS: Particle size distribution of undiluted poractant alfa aerosols generated by a customized eFlow-Neos nebulizer system was determined by laser diffraction. The theoretical nebulized surfactant lung dose was estimated in vitro in a clinical setting replica including a neonatal continuous positive airway pressure (CPAP) circuit, a cast of the upper airways of a preterm neonate, and a breath simulator programmed with the tidal breathing pattern of an infant with mild respiratory distress syndrome (RDS). A dose-response study with nebulized surfactant covering the 100-600 mg/kg nominal dose-range was conducted in RDS-modelling, lung-lavaged spontaneously-breathing rabbits managed with nasal CPAP. The effects of nebulized poractant alfa on arterial gas exchange and lung mechanics were assessed. Exogenous alveolar disaturated-phosphatidylcholine (DSPC) in the lungs was measured as a proxy of surfactant deposition efficacy. RESULTS: Laser diffraction studies demonstrated suitable aerosol characteristics for inhalation (mass median diameter, MMD = 3 µm). The mean surfactant lung dose determined in vitro was 13.7% ± 4.0 of the 200 mg/kg nominal dose. Nebulized surfactant delivered to spontaneously-breathing rabbits during nasal CPAP significantly improved arterial oxygenation compared to animals receiving CPAP only. Particularly, the groups of animals treated with 200 mg/kg and 400 mg/kg of nebulized poractant alfa achieved an equivalent pulmonary response in terms of oxygenation and lung mechanics as the group of animals treated with instilled surfactant (200 mg/kg). CONCLUSIONS: The customized eFlow-Neos vibrating-membrane nebulizer system efficiently generated respirable aerosols of undiluted poractant alfa. Nebulized surfactant delivered at doses of 200 mg/kg and 400 mg/kg elicited a pulmonary response equivalent to that observed after treatment with an intratracheal surfactant bolus of 200 mg/kg. This bench-characterized nebulized surfactant delivery strategy is now under evaluation in Phase II clinical trial (EUDRACT No.:2016-004547-36).

Functional and molecular evidence of myelin- and neuroprotection by thyroid hormone administration in experimental allergic encephalomyelitis.

AIMS: Recent data in mouse and rat demyelination models indicate that administration of thyroid hormone (TH) has a positive effect on the demyelination/remyelination balance. As axonal pathology has been recognized as an early neuropathological event in multiple sclerosis, and remyelination is considered a pre-eminent neuroprotective strategy, in this study we investigated whether TH administration improves nerve impulse propagation and protects axons. METHODS: We followed up the somatosensory evoked potentials (SEPs) in triiodothyronine (T3)-treated and untreated experimental allergic encephalomyelitis (EAE) Dark-Agouti female rats during the electrical stimulation of the tail nerve. T3 treatment started on the 10th day post immunization (DPI) and a pulse administration was continued until the end of the study (33 DPI). SEPs were recorded at baseline (8 DPI) and the day after each hormone/ vehicle administration. RESULTS: T3 treatment was associated with better outcome of clinical and neurophysiological parameters. SEPs latencies of the two groups behaved differently, being briefer and closer to control values (=faster impulse propagation) in T3-treated animals. The effect was evident on 24 DPI. In the same groups of animals, we also investigated axonal proteins, showing that T3 administration normalizes neurofilament immunoreactivity in the fasciculus gracilis and tau hyperphosphorylation in the lumbar spinal cord of EAE animals. No sign of plasma hyperthyroidism was found; moreover, the dysregulation of TH nuclear receptor expression observed in the spinal cord of EAE animals was corrected by T3 treatment. CONCLUSIONS: T3 supplementation results in myelin sheath protection, nerve conduction preservation and axon protection in this animal model of multiple sclerosis.

The Morfeo Study: A 1-year follow-up of complications of vegetative state in a dedicated facility.

OBJECTIVES: Vegetative State (VS) implies significant issues. The aim of the MORFEO study is to identify the most relevant complications in VS patients and to supply clinicians and policy-makers with data derived from the analysis of a cohort of patients treated in a dedicated long-term facility setting. METHODS: A cohort of 22 VS patients treated between 2003 and 2007 were enrolled and followed up for 1 year. The information recorded were: Disability Rating Scale (DRS), Levels of Cognitive Functioning (LCF), pressure sores, nutritional status, neurological complications, articular complications (passive range of motion-ROM), deep-vein thrombosis and infections. The Kolmogorov-Smirnov test was used to verify the normal distribution of the variables. The indicators of complications were analysed with the Friedman test (continuous variables) and with the Cochran Q test (dichotomous variables). RESULTS: DRS and LCF values showed no significant variation. The number of pressure sores decreased. The nutritional status remained satisfying. The ROM worsened in lower limb joints; a trend (p = ns) towards an improved range was observed in shoulders and elbows. Fifteen infections were recorded. CONCLUSIONS: The data that proved significant suggest a minimum set of quality-of-care indicators in VS patients: pressure sores follow-up, nutritional status, ROM and incidence of infections.

CDKL5 knockout leads to altered inhibitory transmission in the cerebellum of adult mice.

Mutations in the X-linked cyclin-dependent kinase-like 5 gene (CDKL5) are associated to severe neurodevelopmental alterations including motor symptoms. In order to elucidate the neurobiological substrate of motor symptoms in CDKL5 syndrome, we investigated the motor function, GABA and glutamate pathways in the cerebellum of CDKL5 knockout female mice. Behavioural data indicate that CDKL5-KO mice displayed impaired motor coordination on the Rotarod test, and altered steps, as measured by the gait analysis using the CatWalk test. A higher reduction in spontaneous GABA efflux, than that in glutamate, was observed in CDKL5-KO mouse cerebellar synaptosomes, leading to a significant increase of spontaneous glutamate/GABA efflux ratio in these animals. On the contrary, there were no differences between groups in K(+) -evoked GABA and glutamate efflux. The anatomical analysis of cerebellar excitatory and inhibitory pathways showed a selective defect of the GABA-related marker GAD67 in the molecular layer in CDKL5-KO mice, while the glutamatergic marker VGLUT1 was unchanged in the same area. Fine cerebellar structural abnormalities such as a reduction of the inhibitory basket 'net' estimated volume and an increase of the pinceau estimated volume were also observed in CDKL5-KO mice. Finally, the BDNF mRNA expression level in the cerebellum, but not in the hippocampus, was reduced compared with WT animals. These data suggest that CDKL5 deletion during development more markedly impairs the establishment of a correct GABAergic cerebellar network than that of glutamatergic one, leading to the behavioural symptoms associated with CDKL5 mutation.

Uso de tarjetas de medicación en el manejo de estado epiléptico en un servicio de emergencias. Mejoras en prescripción, preparación y administración de drogas / Use of medication cards in the management of statu epilepticus at an emergency department. Improvements in drug prescription, preparation and administration

Introducción: Los errores de medicación (EM) causan una elevada morbimortalidad y generan costos innecesarios. El servicio de emergencias (SE) presenta un mayor riesgo de EM que otras áreas. El desarrollo de una herramienta que estandarice el uso de fármacos podría mejorar la seguridad y el proceso de medicación. Objetivos: Evaluar las mejoras en el proceso de medicación mediante el uso de tablas de medicación (TM) durante la atención del estado epiléptico (EE). Materiales y métodos: Se realizó un estudio de tipo antes y después no controlado. La intervención fue el desarrollo e implementación de TM. Se relevó in situ la prescripción, preparación y administración de fármacos incluidos en las TM durante segunda quincena de Octubre y mes de Noviembre 2016, previo a la implementación de la herramienta, y en el mismo período de 2017, luego de la inducción e implementación de las tablas. Se registraron los EM y se categorizaron de acuerdo a la etapa del proceso en que ocurrieron. Resultados: En el período pre-intervención se realizaron 14 registros, 86% (12) tenía al menos un error; 57% (8) errores en la etapa de prescripción, 57% (8) en la de preparación y 21% (3) en la de administración. En el período post-intervención se realizaron 17 registros, 12% (2) tenía por lo menos un EM. No se registraron errores en la fase de prescripción, hubo 12% (2) de errores de preparación y 6% (1) de administración. Conclusión: La implementación de las TM para la estandarización del uso de fármacos en EE resultó una medida muy positiva, mejorando la seguridad en el proceso de medicación (AU)

Introduction: Medication errors (ME) are associated with high morbidity mortality and lead to unnecessary costs. The risk of ME is higher at the emergency department (ED) than in other areas. Developing a tool that standardizes drug use may improve safety and medication processes. Objectives: To evaluate improvements in the medication process by using medication cards (MCs) during status epilepticus (SE) care. Materials and methods: An uncontrolled before-and-after study was conducted. The intervention was the development and implementation of MCs. The in situ prescription, preparation, and administration of drugs included in the MCs was recorded during the second half of October and November 2016, prior to the implementation of the tool, and in the same period of 2017, after the introduction and implementation of the MCs. ME were recorded and categorized according to the stage of the process in which they occurred. Results: In the pre-intervention period 14 episodes were recorded; in 86% (12) at least one error occurred; 57% (8) were ME in the prescription stage, 57% (8) were ME in the preparation stage, and 21% (3) were ME in the administration stage. In the post-intervention period 17 errors were recorded, in 12% (2) at least one ME occurred. No errors were recorded in the prescription stage, 12% (2) were preparation errors, and 6% (1) administration errors. Conclusion: The implementation of MCs for the standardization of medications used in the RU was successful, improving safety in the medication process (AU)

CHF5074 (CSP-1103) induces microglia alternative activation in plaque-free Tg2576 mice and primary glial cultures exposed to beta-amyloid.

Activation of microglia associated with neuroinflammation and loss of phagocytic activity is considered to play a prominent role in the pathogenesis of Alzheimer's disease (AD). CHF5074 (CSP-1103) has been shown to improve cognition and reduce brain inflammation in patients with mild cognitive impairment (MCI). CHF5074 was also found to reverse impairments in recognition memory and improve hippocampal long-term potentiation when administered to plaque-free Tg2576 mice (5-month-old) for 4 weeks. Though, no investigation has focused on the consequence of CHF5074 treatment on microglia polarization yet. In this study we evaluated the effect of CHF5074 administration (375 ppm in the diet) to 5-month-old Tg2576 mice on the expression of pro-inflammatory (M1) genes, Interleukin 1 beta (IL-1ß), Tumor Necrosis Factor alpha (TNFα) and inducible Nitric Oxide Synthase (iNOS), and anti-inflammatory/phagocytic (M2) markers Mannose Receptor type C 1 (MRC1/CD206), Triggering Receptor Expressed on Myeloid cells 2 (TREM2) and Chitinase 3-like 3 (Ym1). No changes of pro-inflammatory gene transcription but a reduced expression of MRC1/CD206, TREM2 and Ym1 were detected in the hippocampus of young Tg2576 mice receiving normal diet, when compared to wild-type littermates. CHF5074 did not affect the pro-inflammatory transcription but significantly increased the expression of MRC1/CD206 and Ym1. CHF5074 effects appeared to be hippocampus-specific, as the M2 transcripts were only slightly modified in the cerebral cortex. In primary cultures of mouse astrocyte-microglia, CHF5074 totally suppressed the expression of TNF-α, IL-1ß and iNOS induced by 10 µM ß-amyloid1-42 (Aß42). Moreover, CHF5074 significantly increased the expression of anti-inflammatory/phagocytic markers MRC1/CD206 and TREM2, reduced by the Aß42 application alone. The effect of CHF5074 was not reproduced by ibuprofen (3 µM or 500 µM) or R-flurbiprofen (3 µM or 100 µM), as both compounds limited the pro-inflammatory gene expression but did not modify the anti-inflammatory/phagocytic transcription. These data show that CHF5074 specifically drives the expression of microglia M2 markers either in young Tg2576 hippocampus or in primary astrocyte-microglia cultures, suggesting its potential therapeutic efficacy as microglial modulator in the early phase of AD.

N-nitrosoproline excretion in the presence and absence of gastric disease.

N-nitrosoproline (NPRO) excretion, an indicator of endogenous nitrosation, was measured in a group of hospital inpatients who were identified by endoscopy and gastric biopsy as either having gastric lesions or having healthy stomachs. NPRO was assayed in background 24-hour urine samples and samples collected after loading doses of nitrate and L-proline. The presence of gastric lesions was associated with altered gastric pH and concomitant changes in gastric juice nitrate and nitrite concentration. Gastric juice pH increased with increasing severity of gastric disease (P = 0.031) and patients with normal stomachs had a lower gastric pH than those with chronic atrophic gastritis (CAG) (3.0 vs. 6.5, P = 0.017). The changes in gastric juice nitrate concentration were in the reverse direction (P = 0.002 for trend) with normal patients having higher mean levels than CAG patients (12.7 vs. 5.5 micrograms/ml, P less than 0.0001). Nitrite concentration increased with severity of gastric disease but the results were not significant (normal, 82.9 vs. CAG, 223.4 ng/ml, P = 0.069). No association was found between the presence of gastric lesions and increased urinary NPRO excretion. Mutagenic activity was not detected in any of the gastric juice samples.

Uncommon intrathoracic extrapulmonary tumor: primary hemangiopericytoma.

We report an unusual case of primary intrathoracic extrapulmonary hemangiopericytoma. Despite the large size and rapid growth of the tumor, no histological sign of malignancy was present. Tumor cells immunostained positively only to vimentin.

Toxic injury to rat gut musculature following intraperitoneal administration of 2-t-butyl-4-methoxyphenol.

The 100-fold increase in toxicity of intraperitoneal (i.p.) rather than orally administered 2-t-butyl-4-methoxyphenol (BHA) is adduced to the depressive effect which this compound exerts on the contractility of the gut musculature. A structure/activity relation study shows the t-butyl group on the benzene ring as being the major determinant of i.p. BHA toxicity. Contractile activity, elicited by field electrical stimulation, acetylcholine or Ba2+, of the ileum longitudinal muscle preparation from BHA-treated rats was greatly reduced 30 min after i.p. injection, and almost absent during the subsequent 48 h. Electron-microscope examination of ileum longitudinal muscle also showed partial destruction of cell membranes 4 h after BHA administration with subsequent mitochondrial swelling and destruction of cristae, myofibrillar fragmentation and cell necrosis. Comparable suppression of contractile activity and morphological damage were observed in BHA or t-butylbenzene incubated ileum segments where longitudinal smooth muscle contractility was irreversibly depressed in a time- and dose-dependent manner. These convergent findings point to the toxic effect of i.p. BHA on gut musculature with consequent impairment of intestinal transit.

Early diagnosis of adult respiratory distress syndrome: an experimental model of complement-mediated pulmonary injury.

This article presents an experimental model of pulmonary injury resembling subclinical human adult respiratory distress syndrome (ARDS) mediated by leukocytes. The activation of complement was prolonged by an intraperitoneal injection of a suspension of zymosan in paraffin. The first step in the development of the disease was an accumulation of polymorphonuclear leukocytes in the lungs. No significant changes were observed on chest radiographs or computed tomograms and the pathophysiologic changes were only minimal in spite of the characteristic structural changes. The criteria currently used for the diagnosis of ARDS are not sensitive enough to detect the subclinical phase of the disease.

[A case of macroamylasemia]. / Un caso di macroamilasemia.

An observed case of macroamylasaemia is presented with a description of the identification technique employed. As in cases reported in the literature, antiglobulin sera did not precipitate the complexes. This would appear to exclude the possibility of the immunity-related aetiopathogenesis the patient's immunological and electrophoretic patterns suggested.

[The origin of the arteria sigmoidea ima]. / Sull'insorgenza dell'a. sigmoidea ima.

The authors report the results of their observations on the lowest sigmoid artery, carried out on 92 anatomical specimens coming from surgical operations on the sigmoid and rectum. The specimens were injected and radiographed immediately after their removal . The observations were often preceded by pre-operative angiographies performed for diagnostic reasons. The AA. document all the variational patterns of this artery. They confirm the poor functional value of this artery more known by the old surgeons than by the anatomists.

[Surgical treatment of carcinoma of the pancreas]. / Sul trattamento chirurgico del carcinoma del pancreas.

Personal experience acquired in the use of radical and palliative techniques in the treatment of 18 cases of carcinoma of the pancreas and 1 of carcinoma of the ampulla hepatopancreatica is reported. The results show that duodenocephalo-pancreasectomy is a sound procedure, particularly in the absence of lymph node invasion. Operative mortality is low if the surgeon is experienced. In the even of serious and prolonged cholostasis, the operation can be preceded by a biliary shunt. According to the Authors there are not yet valid reasons as to the survival to prefer total pancreasectomy instead the partial pancreasectomy because of the high mortality of the former. Early diagnosis and the recognition of precancerous conditions still represent determining factors as far as operability and hence survival are concerned. The efficacy of chemotherapy is uncertain at present.

[Spontaneous bilio-digestive fistula of lithiasic origin]. / In tema di fistole bilio-digestive spontanee a genesi litiasica.

Personal experience about 11 cases of spontaneous biliary enteric fistula caused by lithiasic disease operated in the General Surgical Clinic of the University of Siena from 1969 to 1988, is reported. The epidemiology, pathogenesis, symptomatology, indications for operation and various operating techniques performed are discussed. In the gallstone intestinal obstruction, the authors prefer to resolve immediately the obstruction, deferring to a later moment the specific operation for the treatment of the biliary enteric fistula and the lithiasic disease.

[Acute volvulus of the sigmoid colon]. / Considerazioni sul volvolo acuto del sigma.

Seventeen cases of acute volvulus of the sigmoid colon are presented. Comparison with other series showed that the mortality rate for fixation with or without a shunt are more or less the same. Excellent results were obtained with the Paul Mikulicz method. An intention to use this method in all cases in the future, irrespective of the patient's local and general condition, is expressed.

[Antibiotic prophylaxis with ++vancomycin in corrective surgery with alloplastic material]. / Antibioticoprofilassi con vancomicina nella chirurgia riparativa con materiale alloplastico.

The authors report a controlled series of 40 patients treated for recidivating inguinal hernia and laparocele with the implant of alloplastic material. Short-term prophylaxis with vancomycin was performed in all cases. The antibiotic was found to be well tolerated at the doses used and this was associated with a good clinical success rate. The use of vancomycin in prophylaxis for this type of surgery is considered a rational choice and the use of short-term prophylaxis reduces the risks linked to possible collateral effects.

[Gasless laparoscopic cholecystectomy. Our experience with 130 cases compared with 450 cases treated with the CO2 technique]. / Colecistectomia laparoscopica gasless. La nostra esperienza su 130 casi confrontati con 450 casi trattati con la tecnica con CO2.

Alongside the technique based on the creation of an abdominal cavity for surgery following the introduction of gas (usually CO2) into the peritoneal cavity, a new method has been developed. This involves the use of an atraumatic mechanical lifting device connected to the same abdominal wall (gasless laparoscopy). The authors report a technique that uses an inflatable cushion inserted into the abdomen through a periumbilical incision. The cushion is connected to an external motorized hydraulic jack fixed to the operating table, fitted with an electric motor and friction gear. Between May 1991 and June 1998, 580 patients underwent laparoscopic cholecystectomy. Since December 1995 a total of 130 patients have undergone surgery using gasless laparoscopy. Shoulder pain and pain in the upper abdominal quadrant were no longer reported; pain was present in 70% of the patients operated using the CO2 technique. There was also a marked reduction in the anesthesiological risks, above all in elderly patients with cardiopulmonary insufficiency. Surgical manoeuvres are made easier owing to the possibility of using traditional surgical instruments. Washing and continuous aspiration allow a good control of intraoperative hemostasis, and reduce the phenomenon of lens misting without the risk of losing pneumoperitoneum. Less visibility of the surgical field was reported, particularly in obese patients, above all because of the reduced diaphragmatic distension and the lack of displacement of the intestinal loops. In the authors' opinion the gasless technique is suitable above all in patients affected by cardiopulmonary disorders in whom hypercapnia might represent a significant operating risk.

Clawed toes deformity due to spasticity of the extensor digitorum muscle. A case report.

Spasticity is a characteristic and early component of the lesions of the pyramidal tract of demyelinising diseases such as multiple sclerosis, and is included in the chief neurological scales to evaluate this disease. We report the case of a woman with a clawed toes deformity caused by spasticity of the extensor digitorum (longus and brevis) muscles.