Generation of iPSC lines derived from skin fibroblasts of two healthy controls using non-transmissible form of Sendai Virus.

We established two iPSC lines starting from skin fibroblasts of two healthy individuals using Sendai-virus-based technique. The obtained iPSCs were characterized showing same STR profile as starting fibroblasts, normal karyotype, loss of stemness vectors, expression of stemness markers, both through real-time PCR and immunofluorescence, (OCT4, SOX2, TRA-1-60, NANOG and SSEA4) and in vitro differentiation into three germ layers.

[Measurement of real personal noise attenuation using earplugs with the E-A-Rfit system]. / La misura del reale abbattimento soggettivo di rumore con uso di inserti auricolari tramite sistema E-A-Rfit.

BACKGROUND: The assessment of efficiency of hearing protection devices (HPDs), conducted above statutory limits, must be made using a standardized method while devices are worn; however, standardized and suitable laboratory conditions are difficult to encounter at the workplace. To overcome this problem, there are methods of measurement at the workplace such as "field-microphone-in-real-ear" (F-MIRE). OBJECTIVES: The study was concerned with the measurement of real noise attenuation using earplugs and a new evaluation system: we checked the difference between "real" attenuation (at workplace) and "theorical" attenuation (reproduced in the laboratory) as stated by the manufacturer. METHODS: We used the E-A-Rfit computerized method, which measures the loss of attenuation of earplugs in the ear, calculating the difference of sound pressure between an "outside" microphone and an "inside" one, in relation to the same earplug. The measurements at the workplace were carried out on eight subjects with good hearing levels (aged between 20 and 25 years), who were trained to wear the devices correctly. After the tests carried out with the E-A-Rfit system, which does not require a subjective answer, we obtained graphs and tables showing real noise attenuation. RESULTS/CONCLUSIONS: We propose a comparison between hearing threshold for frequency, personal attenuation rating (PAR) and single number rating (SNR, provided by manufacturer): a difference of 10 dB (PAR 27 db vs. SNR 37 dB) was clearly evident although dissimilar methods were used to obtain such values. The instrument is rapid, simple and objective to use and also allows personalized information and training for every worker.

Generation of an iPSC line from skin fibroblasts of a patient with Joubert syndrome carrying the homozygous loss of function variant c.787dupC in the AHI1 gene.

We produced an iPSC line from a patient with Joubert syndrome carrying the homozygous c.787dupC variant in the AHI1 gene. The iPSC line was obtained by reprogramming skin fibroblasts, mycoplasma-free, using Sendai-virus-based technique. Characterization of iPSCs showed the same Short Tandem Repeats profile than fibroblasts, normal karyotype, expression of staminal markers (OCT4, SOX2, SSEA4 and NANOG) and ability to differentiate into three germ layers in vitro.

[And yet there is a risk--do it yourself]. / Eppur c'è rischio--Il bricolage.

Our study has examined the valuation of exposure to dust, noise and hand-arm vibrations (HAVs) during "extra-professional" activities of do-it-yourself, like wood smoothing, considering a group of 8 heterogeneous volunteer subjects (not expert of the sector), to find any kind of variability among the subjects. The results have shown a moderate risk for dust exposure and a realer one for HAVs, also higher noise exposure levels when an aspiration system is added to the sander. It's important that the exposure time considered in this study is not comparable to professional time exposure, cause of the "domestic" feature of this activities. Moreover, data could be influenced by different use conditions, grip and material grounds. It's also significant that there are not controls, formation and information of the subjects about the health risks, as well as ipersusceptibility.

Characterization of signals for a Divertor Tokamak Test facility interferometer/polarimeter system.

In magnetically confined fusion experiments, laser interferometer/polarimeter systems allow one to determine plasma density, give valuable information on the internal magnetic fields, and contribute to the evaluation of the plasma magnetic equilibrium and to the real-time estimation of the q profile to allow feedback configuration control. This work presents an analysis of the interferometric and polarimetric signals of a multi-chord far-infrared interferometer/polarimeter for the divertor tokamak test facility, the new tokamak device currently under construction in Italy. The polarimetric signals are calculated both with approximate formulas and by solving the equation describing the evolution of the laser beam polarization inside the plasma using the Mueller formalism. The latter method correctly accounts for crosstalk between Faraday rotation and the Cotton-Mouton effect. The impact of the plasma birefringence on the interferometric phase shift is also studied, and it is found that a perturbation of the interferometric phase shift is present also in the case of an initial fixed linear polarization of the probe laser beam.

Spontaneous in-the-bag intraocular lens luxation into the vitreous cavity: last-stage complication of pseudoexfoliative syndrome after phacoemulsification.

PURPOSE: We describe 8 cases of late spontaneous in-the-bag intraocular lens (IOL) luxation into the vitreous cavity, which occurred at the Department of Ophthalmology and Neurosurgery of the University of Siena between January and December 2006. METHODS: In this interventional case series, the medical records of all patients with posterior luxation of in-the-bag IOLs - who had undergone a pars plana vitrectomy with IOL removal and scleral fixation IOL implantation between January and December 2007 at the Department of Ophthalmology and Neurosurgery of Siena, Italy - were retrospectively reviewed. RESULTS: The final post- operative visual acuity was 20/30 or better in 6 patients, while myopic macular degeneration and total retinal detachment limited visual acuity in the remaining 2 patients. CONCLUSION: The high prevalence of pseudoexfoliation (PEX) in the patients who had been operated for cataract phacoemulsification in our department could explain the occurrence of 8 posterior luxations of in-the-bag IOLs in only 1 year. Our study suggests that for the next years we will expect an increase in occurrence of spontaneous in-the- bag IOL luxations in the vitreous cavity. This condition could represent the last stage of PEX syndrome.

Clinical operational tolerance after kidney transplantation: a short literature review.

The clinical era of solid organ transplantation started with a renal transplantation (RT) performed between identical twins in Boston in 1954. The patient did not receive any immunosuppression, thus representing the very first case of operational tolerance (Tol). However, more than half a century later, we must admit the inadequacy of our knowledge regarding such a fundamental aspect of transplant immunology, as demonstrated by the fact that Tol has never been achieved in an intention-to-treat protocol. Herein we aim to shortly review the worldwide experience on clinical operational Tol after RT. Thus far, reports on successful cases of Tol after RT have been anecdotal: the largest series included no more than 10 individuals. We will understand that Tol can develop even in the presence of either HLA mismatches or blood group incompatibility at baseline, in the presence of anti-HLA antibodies during follow-up, as well as in patients having experienced acute rejection. Despite the lack of robust evidence, the fact that Tol is often accidentally discovered by transplant physicians during follow-up in noncompliant patients justifies the hypothesis that the real number of Tol cases might be much higher than currently reported.

Incidence of urinary tract infections caused by germs resistant to antibiotics commonly used after renal transplantation.

BACKGROUND: The inadequate utilization of antibiotics is responsible for the development of urinary tract infections (UTI) after renal transplantation (RT), through the induction of resistance to the antibiotics themselves. The purpose of this study was to evaluate the incidence of resistance to cefotaxime (CEF) and trimethoprim/sulfamethoxazole (TMP-SMX), routinely used for surgical perioperative prophylaxis and prevention of Pneumocystis carinii, respectively. MATERIALS AND METHODS: We enrolled all adult patients having received an RT from 2001 to 2006 and having a minimum follow-up of 6 months. Urine cultures (UC) were routinely performed at every outpatient clinic control and whenever required by the onset of significant clinical signs/symptoms. UTI was diagnosed by the presence of a positive UC. The endpoint of the study was the emergence of bacterial strains resistant to either CEF or TMP/SMX. RESULTS: We recorded 169 UTI in 76 patients (38 men/38 women, 33%) over a mean follow-up of 779.9+/-523.3 days. Thirty-nine patients (51%) developed more than 1 UTI episode. When gram-negative bacteria were considered, 102/144 (70.8%) tests showed resistance to TMP/SMX, while data were available in about only 7 gram-positive infections (5/7, 71%). CEF was tested less frequently with 21/43 (49%) germs resistant to this molecule. CONCLUSIONS: The onset of bacterial resistance to either TMP/SMX or CEF is frequent after RT. A wiser stricter utilization of antibiotics is mandatory. Standard antibiotic protocols should be revised.

De novo autoimmune hepatitis following liver transplantation: a case report.

De novo autoimmune hepatitis (AIH), a rare disorder first described in 1998, appears in patients with liver transplants due to autoimmune and nonautoimmune etiologies. De novo AIH occurs in 2.5% to 3.4% of allografts; children seem to have a predilection for this syndrome. We have present herein a case of a liver allograft recipient who developed chronic hepatitis associated with autoimmune features outlining the clinical course, liver histology, and response to treatment.

Risk factors that can influence kidney transplant outcome.

The survival and function of a kidney transplant are influenced by numerous immunological and nonimmunological factors. The aim of this study was to evaluate the role of a number of cadaveric donor parameters on transplanted kidney function, and in particular on the occurrence of delayed graft function (DGF) since DGF is one of the most important factors in long-term organ survival. This study looked at 143 patients who underwent kidney transplant of whom 32 displayed DGF. The creatinine levels in organ recipients, which were evaluated during a follow-up that ranged between 6 months and 4 years, were significantly higher among recipients who developed DGF after transplant (1.8 +/- 0.7 vs 1.4 +/- 0.4; P = .02). The following donor parameters were taken into consideration: history of diabetes and hypertension; creatinine levels; inotropie therapy; problems relating to hemodynamics (hypotension and/or cardiac arrest); and cold ischemia time. We observed that a donor history of hypertension (46.8% DGF vs 23.27% no DGF; P = .01) and high levels of donor creatinine prior to organ removal (1.9 +/- 1.2 mg/dL DGF vs 1.2 +/- 0.9 mg/dL no DGF; P = .007) were significant risk factors for DGF among kidney recipients. No significant differences were found for others factors between recipients with versus without DGF.

Conservative treatment of keratoconus by riboflavin-uva-induced cross-linking of corneal collagen: qualitative investigation.

PURPOSE: To assess corneal tissue modifications after riboflavin-UVA-induced cross-linking of corneal collagen in patients with progressive keratoconus as well as regeneration of epithelium and subepithelial nerve plexus by in vivo HRT II system confocal microscopy in humans. METHODS: Ten patients with progressive keratoconus were treated by riboflavin-UVA-induced cross-linking of corneal collagen, involving assessment of ultrastructural modifications of the corneal epithelium and subepithelial nerve plexus by HRT II system confocal microscopy. Treatment included instillation of 0.1% riboflavin-20% dextrane solution 5 minutes before UVA irradiation and every 5 minutes for a total of 30 minutes. Radiant energy was 3 mW/cm 2 or 5.4 Joule/cm 2 and the source was dual UVA (370 nm) light-emitting LED. The protocol included the operation followed by antibiotic medication and eye dressing with a soft therapeutic contact lens. Changes in epithelium and subepithelial and stromal nerve plexus were assessed by HRT II system confocal microscopy in vivo. RESULTS: After 5 days of soft contact lens wearing, corneal epithelium has a regular morphology and density. Disappearance of subepithelial stromal nerve fibers was observed in the central irradiated area where, 1 month after the operation, initial reinnervation was microscopically observed. No changes in nerve fibers were observed in the peripheral untreated with a clear lateral transition between the two areas. Six months after the operation, the anterior subepithelial stroma was recolonized by nerve fibers with restoration of corneal sensitivity. CONCLUSIONS: HRT II system confocal microscopy confirms corneal epithelium restore and re-innervation after riboflavin-UVA-induced collagen cross-linking directly in vivo in humans.

[Parathyroidectomy: bilateral exploration of the neck vs minimally invasive radioguided treatment]. / Paratiroidectomia: esplorazione bilaterale del collo versus trattamento radio-guidato mini-invasivo.

AIM: In the therapy of primary hyperparathyroidism, the first surgical intervention, if efficacious, can remarkably reduce the incidence of persistence and relapses which are approximately about 5%. At present, the surgical approach of choice should involve the bilateral exploration of the neck. METHODS: In the light of the high sensibility (91%) and specificity (98.8%) in the localization of parathyroid adenomas obtained by the parathyroid 99mTc-MIBI scintigraphy, we submitted, prospective and at random, between January 2001 and July 2004, 69 patients with primary hyperparathyroidism, to a conventional surgical treatment (bilateral exploration of the neck: 35 patients) or minimally-invasive approach (minimally invasive radioguided parathyroidectomy: 34 patients). This method consists of the injection of 50 mCi of 99mTc Sestamibi 2 h before the operation and the execution of parathyroid scintigraphy. When the adenoma is evident, we perform an incision of about 4 cm in the neck, 2 cm over the jugulum and the surgical dissection is guided by a probe showing the emission of gamma rays. RESULTS: The parameters considered in order to compare the 2 groups, i.e. operating time, hospital stay and time of recovery were reduced in a significant way in the group submitted to the minimally invasive radioguided parathyroidectomy (MIRP). There were no complications in the 2 groups. In the follow-up we did not observe cases of persistence or relapses. CONCLUSIONS: Therefore, we can confirm that the minimally invasive radioguided parathyroidectomy is a safe and efficacious method as well as the bilateral exploration of the neck. Moreover, cost reduction may convince many surgeons to consider MIRP the <> in the management of primary hyperparathyroidism.

[Silicone occlusive sheeting vs silicone cushion for the treatment of hypertrophic and keloid scars. A prospective-randomized study]. / Bendaggio occlusivo al silicone vs cuscino al silicone nel trattamento delle cicatrici ipertrofiche e cheloidee. Studio prospettico-randomizzato.

BACKGROUND: Silicone gel and silicone occlusive sheeting are widely used at present for the treatment of hypertrophic and keloid scars. In recent studies the possibility was raised that static electricity generated by friction activated silicone sheeting could be the reason for this effect, and that it can, with time, cause involution of hypertrophic and keloid scars. Objective of this study was to test this hypothesis and to observe weather a continuous and also an increased negatively charged static electric field will shorten the treatment period. A silicone cushion was developed with the purpose of increasing a negative static-electric charge to accelerate the regression process. METHODS: From November 2001 to June 2002 we studied in a prospective randomized study, 72 patients with hypertrophic an keloid scars. The trial extended over a 8-month period. 37 patients underwent silicone occlusive sheeting, the remaining 35 patients underwent silicone cushion (Clinicel). RESULTS: Treatment with the silicone cushions yielded 74,2% cessation of itching and burning followed by pallor and flattening of the scar, some markedly so, over a few weeks to 5 months period. Additional 25,7% had their scars resolved in up to 8 months of treatment. Four patients (11,4%) who add recalcitrant scars with little response to the use of the silicone cushion were given intralesional corticosteroid injections, in addiction to the use of the cushion, resulting in a fairly rayed resolution of these scars over a period of 2 months. Treatment with the silicone occlusive sheeting yielded 52,3% itching and burning cessation followed by pallor and flattening of the scar, some markedly so, over a few weeks to 5 months period. Additional 22,1% had their scars resolved in up to 8 months of treatment. In conclusion by comparing the results of this trial using silicone cushions for the treatment of hypertrophic and keloid scars with those obtained using silicone gel or occlusive sheeting, a much faster response was demonstrated.

Paying Attention to Compassion Fatigue in Emergency Nurses.

Symptom recognition, institutional support, and healthy coping strategies are crucial.

Changes in the blood coagulation, fibrinolysis, and cytokine profile during laparoscopic and open cholecystectomy.

BACKGROUND: It has long been known that a hypercoagulability state develops after surgery. A surge in circulating cytokine levels is also commonly found in the postoperative period. These cytokines have all been shown to be capable of inducing a hypercoagulability state. Recently laparoscopic cholecystectomy (LC) has been introduced, and its advantages over the open procedure seem related to the reduced surgical trauma. LC is associated with a diminished acute-phase response compared with the open procedure. Our present knowledge on the influence of laparoscopic upon coagulation and fibrinolysis is incomplete and based on a few studies. METHODS: The aim of this prospective, nonrandomized study was to investigate hemostatic system alterations in patients who undergo open and laparoscopic cholecystectomy. In addition we also measured the plasma cytokine profile to explore any relationship between changes in plasma cytokine levels and postoperative coagulation profile. Between September 1999 and April 2002, 71 patients were nonrandomly assigned to open (group 1) or laparoscopic cholecystectomy (group 2). All patients from group 1 were operated by a surgical team different from ours, who prefers the OC procedure. The patients with acute cholecystitis were excluded. Prothrombin fragment 1.2 (F1.2), thrombin-antithrombin (TAT), fibrinogen, soluble fibrin, antithrombin III (AT), protein C, plasminogen, and D-dimer levels were measured at baseline and at 1, 24, 48, and 72 h postoperatively. Serial serum levels of IL-1beta and IL-6 were measured by colorimetric enzyme-linked immunosorbent assay (ELISA). RESULTS: Plasma levels of F1.2, TAT, fibrinogen, soluble fibrin, and D-dimer increased significantly in group 1. Plasma levels of AT, protein C, and plasminogen decreased in both groups. In the OC group, the serum IL-3 and IL-6 levels began to significantly increased as early as 1 h from the beginning of the operation, revealing a peak at the sixth hour. When IL-6 and IL-1 levels were markedly elevated also, F1.2, fibrinogen, and soluble fibrin levels were increased. CONCLUSIONS: Only mild hypercoagulability was observed in patients who had undergone laparoscopic cholecystectomy. The cytokine surge was correlated with hypercoagulability. There was in fact a positive correlation between IL-6 level and hypercoagulability. The correlation between cytokine levels and coagulation activation may be related to the type of surgery performed. Further studies are required to investigate these issues.

A comparison of serum interleukin-6 concentrations in patients treated by cholecystectomy via laparotomy or laparoscopy.

BACKGROUND/AIMS: Interleukin-6 (IL-6), a multifunctional cytokine, is expressed by various cells after many stimuli. This cytokine release is related, among other things, to the extent of the surgically-induced trauma. Laparoscopic cholecystectomy (LC) is a so-called "mini-invasive" surgical procedure and, on the basis of this consideration, the aim of the present prospective non-randomized study, is to examine (a) whether the IL-6 is modified and how, in patients after LC compared to patients undergoing open cholecystectomy (OC), (b) whether these findings are indicative of an increased risk to develop infectious complications and whether they are therefore clinically significant. METHODOLOGY: Circulating IL-6 level was measured using a random access chemiluminescense-immunoassay system in 71 patients before the operation (time 0) and 1, 2, 3, 6, 24 and 48 hours after the beginning of the operation. Thirty-five patients underwent OC and 36 LC. RESULTS: The increase in the serum IL-6 during LC was found to be significantly smaller than that during OC and resulted in a smaller extent of postoperative elevations for C-reactive protein. We recorded three cases (8.5%) of postoperative infections in the "open" group and IL-6 concentration normalized only 6 days after surgery. CONCLUSIONS: An increase in the serum IL-6 level during LC is lower in comparison to OC and results in lower postoperative elevation in C-reactive protein. Laparoscopic surgery, associated with a small skin incision and the avoidance of open laparotomy, can thus minimize surgical stress, and provide more favorable postoperative conditions for patients. Indeed excessive and prolonged post-injury elevations are associated with increased morbidity.

Pulsed vs continuous light accelerated corneal collagen crosslinking: in vivo qualitative investigation by confocal microscopy and corneal OCT.

PURPOSE: To assess qualitative corneal changes and penetration of pulsed and continuous light accelerated crosslinking by in vivo confocal microscopy and corneal OCT. METHODS: A total of 20 patients affected from progressive keratoconus were enrolled in the study. Ten eyes of 10 patients underwent an epithelium-off pulsed-light accelerated corneal collagen crosslinking (PL-ACXL) by the KXL UV-A source (Avedro Inc.) with 8 min (1 s on/1 s off) of UV-A exposure at 30 mW/cm(2) and energy dose of 7.2 J/cm(2); 10 eyes of 10 patients underwent an epithelium-off continuous-light accelerated corneal collagen crosslinking (CL-ACXL) at 30 mW/cm(2) for 4 min. Riboflavin 0.1% dextran-free plus hydroxyl-propyl-methylcellulose solution (VibeX Rapid, Avedro Inc.) was used for a 10-min corneal soaking. Treated eyes were examined by in vivo scanning laser confocal analysis and spectral anterior segment OCT at 1, 3, and 6 months. RESULTS: Epithelial stratification and nerves regeneration improved in time, being complete at month 6 in both groups without endothelial damage. Keratocyte apoptosis in PL-ACXL was estimated at a mean depth of ∼200 µm, whereas an uneven demarcation line was detectable by confocal microscopy at a mean depth of 160 µm in CL-ACXL. CONCLUSION: In vivo confocal microscopy and corneal OCT allowed a precise qualitative analysis of the cornea after epithelium-off PL-ACXL and CL-ACXL treatments. Apoptotic effect was higher in pulsed than in continuous light treatments, exceeding 200 µm in corneal stroma. According to different morphological data, the clinical efficacy of ACXL needs to be determined in a long-term follow-up and large cohort of patients.

A microwave interferometer for small and tenuous plasma density measurements.

The non-intrusive density measurement of the thin plasma produced by a mini-helicon space thruster (HPH.com project) is a challenge, due to the broad density range (between 10(16) m(-3) and 10(19) m(-3)) and the small size of the plasma source (2 cm of diameter). A microwave interferometer has been developed for this purpose. Due to the small size of plasma, the probing beam wavelength must be small (λ = 4 mm), thus a very high sensitivity interferometer is required in order to observe the lower density values. A low noise digital phase detector with a phase noise of 0.02° has been used, corresponding to a density of 0.5 × 10(16) m(-3).

Interlamellar cohesion after corneal crosslinking using riboflavin and ultraviolet A light.

AIMS: Collagen crosslinking treatment of progressive keratoconus using the photosensitiser riboflavin and ultraviolet A light of 370 nm wavelength has been shown to increase significantly the tensile strength of corneal collagen by about 300%. In keratoconus, interlamellar and interfibrillar slippage have been proposed as pathogenetic mechanisms. Therefore, the aim of this study was to assess the impact of collagen crosslinking on the interlamellar cohesive force. METHODS: 72 post mortem porcine eyes were divided into six different treatment groups: the untreated control group, the standard crosslinking group, the hypo-osmolar crosslinking group, the stromal swelling group, the formaldehyde group and the α-amylase group. An anterior 9×4 mm strip of 400 µm thickness was prepared using a lamellar rotating microkeratome. For interlamellar cohesive force measurements a splitting plane was created at 50% depth. Force-distance profiles were recorded using a microcomputer-controlled biomaterial testing machine. RESULTS: The mean interlamellar cohesive force was 0.24 N/mm in the untreated control group, 0.26 N/mm in the standard crosslinking group, 0.25 N/mm in the hypo-osmolar crosslinking group, 0.23 N/mm in hydrated corneas, 0.27 N/mm in the formaldehyde group without statistically significant difference. Only the values of the α-amylase group were statistically significantly lowered by 31.5% to 0.16 N/mm. CONCLUSIONS: Surprisingly, corneal crosslinking does not increase the interlamellar cohesive force. In the α-amylase group the cohesive force was mainly decreased because of the digestion of proteoglycans. Crosslinking seems to stabilise only inter- and intrafibrillar, but not interlamellar cohesion.