Electrocardiogram analysis in Anderson-Fabry disease: a valuable tool for progressive phenotypic expression tracking.

Background: Electrocardiogram (ECG) has proven to be useful for early detection of cardiac involvement in Anderson-Fabry disease (AFD); however, little evidence is available on the association between ECG alterations and the progression of the disease. Aim and Methods: To perform a cross sectional comparison of ECG abnormalities throughout different left ventricular hypertrophy (LVH) severity subgroups, providing ECG patterns specific of the progressive AFD stages. 189 AFD patients from a multicenter cohort underwent comprehensive ECG analysis, echocardiography, and clinical evaluation. Results: The study cohort (39% males, median age 47 years, 68% classical AFD) was divided into 4 groups according to different degree of left ventricular (LV) thickness: group A ≤ 9 mm (n = 52, 28%); group B 10-14 mm (n = 76, 40%); group C 15-19 mm (n = 46, 24%); group D ≥ 20 mm (n = 15, 8%). The most frequent conduction delay was right bundle branch block (RBBB), incomplete in groups B and C (20%,22%) and complete RBBB in group D (54%, p < 0.001); none of the patients had left bundle branch block (LBBB). Left anterior fascicular block, LVH criteria, negative T waves, ST depression were more common in the advanced stages of the disease (p < 0.001). Summarizing our results, we suggested ECG patterns representative of the different AFD stages as assessed by the increases in LV thickness over time (Central Figure). Patients from group A showed mostly a normal ECG (77%) or minor anomalies like LVH criteria (8%) and delta wave/slurred QR onset + borderline PR (8%). Differently, patients from groups B and C exhibited more heterogeneous ECG patterns: LVH (17%; 7% respectively); LVH + LV strain (9%; 17%); incomplete RBBB + repolarization abnormalities (8%; 9%), more frequently associated with LVH criteria in group C than B (8%; 15%). Finally, patients from group D showed very peculiar ECG patterns, represented by complete RBBB + LVH and repolarization abnormalities (40%), sometimes associated with QRS fragmentation (13%). Conclusions: ECG is a sensitive tool for early identification and long-term monitoring of cardiac involvement in patients with AFD, providing "instantaneous pictures" along the natural history of AFD. Whether ECG changes may be associated with clinical events remains to be determined.

Art for improving skills in medical education: the validation of a scale for measuring the Visual Thinking Strategies method.

BACKGROUND: The skills needed in the medical and nursing field are considered both for the cognitive and the personal and interpersonal aspects. There are many studies that suggest using artistic practices and pedagogical methods such as Visual Thinking Strategies (VTS) or Artful Thinking in the medical education. The main aim of this research is to validate a grid to evaluate impact of art activities for improving skills in medical education sector. METHODS: The VTSkill grid was created by research group of Sapienza University, selecting the relevant dimension on the basis of literature analysis. To evaluate the validity and reliability, the grid was used in a quasi-experimental study involving the pediatric ward personnel, the nursing and medicine course students of Sapienza University of Rome. This analytic rubric was used to evaluate the written assessment form, composed by open-ended basic question related to the VTS method, administered in association with two images, a work of art and a clinical image. The Number of responders of the validation study was 105. RESULTS: Although obtained from a small sample, both construct validity and reliability analysis showed coherent and statistically significant results. On one hand, the construct validity results showed a relationship path consistent with the hypothesised one derived from previous literature, with relevant p-values (n = 78). On the other hand, the VTSkill reliability was first analysed through the inter-rater evaluation data. This reliability coefficient showed a high degree of convergence of judgments between different evaluators on both image data (n = 55), with statistically significant values ranging from good (r = .77) up to excellent for objectively observable items (r = 1). Similarly, the test-retest reliability coefficients calculated for both clinical and artwork image data resulted statistically significant (n = 95), although ranging from weak to adequate entity (up to r = .77). CONCLUSIONS: Taking into account the high degree of coherence and the stability of measurement of VTSkill in combination with its consistent construct validity, this study suggest the opportunity to implement this measurement tool to research the effect of VTS protocol in future investigations on the field. Therefore, the results of this study will constitute the basis to collect further evidences on how arts-based learning methods can contribute in medical education to improve skills suitable to the health professionals.

Imbibition of Femtoliter-Scale DNA-Rich Aqueous Droplets into Porous Nylon Substrates by Molecular Printing.

This work presents the first reported imbibition mechanism of femtoliter (fL)-scale droplets produced by microchannel cantilever spotting (µCS) of DNA molecular inks into porous substrates (hydrophilic nylon). Differently from macroscopic or picoliter droplets, the downscaling to the fL-size leads to an imbibition process controlled by the subtle interplay of evaporation, spreading, viscosity, and capillarity, with gravitational forces being quasi-negligible. In particular, the minimization of droplet evaporation, surface tension, and viscosity allows for a reproducible droplet imbibition process. The dwell time on the nylon surface permits further tuning of the droplet lateral size, in accord with liquid ink diffusion mechanisms. The functionality of the printed DNA molecules is demonstrated at different imbibed oligonucleotide concentrations by hybridization with a fluorolabeled complementary sequence, resulting in a homogeneous coverage of DNA within the imbibed droplet. This study represents a first step toward the µCS-enabled fabrication of DNA-based biosensors and microarrays into porous substrates.

Inflammation and ventricular-vascular coupling in hypertensive patients with metabolic syndrome.

BACKGROUND AND AIMS: Metabolic syndrome (MetS) is currently considered to raise the risk for type 2 diabetes and cardiovascular events. It has been suggested that part of this risk excess may be due to a cluster of additional factors associated with MetS. We aimed to investigate the role of inflammation on the ventricular-vascular coupling in patients with MetS. METHODS AND RESULTS: We enrolled a total of 227 hypertensive patients (106 with MetS and 121 without MetS) matched for age and gender. Aortic pulse wave velocity (aPWV), intima-media thickness (IMT) and high sensitivity C-reactive protein (CRP) increased according to the number of MetS components. Patients with MetS showed increased aPWV (11.5 ± 3.7 vs. 10.3 ± 2.5 m/s, P = 0.03) compared with controls. In a model adjusted for age, sex, heart rate and mean blood pressure, aPWV resulted increased in patients with CKD (beta 1.29 m/s, 95%CI 0.61-1.96 m/s, P < 0.001) and MetS (beta 0.89 m/s, 95%CI 0.28-1.51 m/s, P = 0.005). After additional adjustment for CRP and IMT, the slope of aPWV was respectively reduced by 16% and 62%, suggesting that inflammation and intima-media thickening could contribute to aortic stiffening in patients with MetS. In these patients, aPWV was also associated with left-ventricular mass index (beta 0.79 g/m2.7, 95%CI 0.05-1.52 g/m2.7, P = 0.05). CONCLUSION: MetS is characterized by an inflammation-dependent acceleration in cardiovascular ageing. This pattern of pathophysiological abnormalities may contribute to amplify the burden of cardiovascular risk in patients with MetS.

Detecting familial hypercholesterolemia by serum lipid profile screening in a hospital setting: Clinical, genetic and atherosclerotic burden profile.

BACKGROUND AND AIMS: Familial hypercholesterolemia (FH) is underdiagnosed and public cholesterol screening may be useful to find new subjects. In this study, we aim to investigate the prevalence of FH patients in a hospital screening program and evaluate their atherosclerotic burden using intima-media thickness (IMT). METHODS AND RESULTS: We screened 1575 lipid profiles and included for genetic analysis adults with a low-density lipoprotein (LDL) cholesterol >190 mg/dL and triglycerides <200 mg/dL and first-degree child relatives with LDL cholesterol >160 mg/dL and triglycerides <200 mg/dL. The diagnosis of FH was presumed by Dutch Lipid Clinic Network (DLCN) criteria and confirmed by the presence of the genetic variant. Mean common carotid intima-media thickness (IMT) was assessed using consensus criteria. After confirming LDL cholesterol value and excluding secondary hypercholesterolemia, 56 subjects with a DLCN ≥4 performed genetic analysis. Of these, 26 had an FH genetic variant. The proportion of patients with a mutation having a DLCN score of 6-8 was 75%; in individuals with a DLCN score >8 it was 100%. Mean IMT was higher in FH patients compared to non FH (0.73 [0.61-0.83] vs 0.71 [0.60-0.75] mm, p < 0.01). Moreover, we detected two mutations not previously described. Finally, simple regression analysis showed a correlation of IMT with LDL cholesterol >190 mg/dL and corneal arcus (p < 0.01 and p < 0.001, respectively). CONCLUSIONS: A hospital screening was useful to detect FH subjects with increased atherosclerosis. Also, next-generation sequencing was able to detect new FH mutations.

Microgel assisted Lab-on-Fiber Optrode.

Precision medicine is continuously demanding for novel point of care systems, potentially exploitable also for in-vivo analysis. Biosensing probes based on Lab-On-Fiber Technology have been recently developed to meet these challenges. However, devices exploiting standard label-free approaches (based on ligand/target molecule interaction) suffer from low sensitivity in all cases where the detection of small molecules at low concentrations is needed. Here we report on a platform developed through the combination of Lab-On-Fiber probes with microgels, which are directly integrated onto the resonant plasmonic nanostructure realized on the fiber tip. In response to binding events, the microgel network concentrates the target molecule and amplifies the optical response, leading to remarkable sensitivity enhancement. Moreover, by acting on the microgel degrees of freedom such as concentration and operating temperature, it is possible to control the limit of detection, tune the working range as well as the response time of the probe. These unique characteristics pave the way for advanced label-free biosensing platforms, suitably reconfigurable depending on the specific application.

Optical fiber tip templating using direct focused ion beam milling.

We report on a method for integrating sub-wavelength resonant structures on top of optical fiber tip. Our fabrication technique is based on direct milling of the glass on the fiber facet by means of focused ion beam. The patterned fiber tip acts as a structured template for successive depositions of any responsive or functional overlay. The proposed method is validated by depositing on the patterned fiber a high refractive index material layer, to obtain a 'double-layer' photonic crystal slab supporting guided resonances, appearing as peaks in the reflection spectrum. Morphological and optical characterizations are performed to investigate the effects of the fabrication process. Our results show how undesired effects, intrinsic to the fabrication procedure should be taken into account in order to guarantee a successful development of the device. Moreover, to demonstrate the flexibility of our approach and the possibility to engineering the resonances, a thin layer of gold is also deposited on the fiber tip, giving rise to a hybrid photonic-plasmonic structure with a complementary spectral response and different optical field distribution at the resonant wavelengths. Overall, this work represents a significant step forward the consolidation of Lab-on-Fiber Technology.

SILS extraperitoneal radical prostatectomy.

AIM: This paper presents the surgical technique for video-laparoscopic extraperitoneal radical prostatectomy carried out through a single incision of 2.5 cm below the navel. METHODS: The technique involves the placement of a port of a new concept, the SILS-port, which has three channels through which you can insert instruments and camera. The study results were evaluated from the surgical, oncological and functional point of view. Ten patients undergoing prostatectomy were compared with the last 10 patients treated at our facilities with increasingly radical prostatectomy video-laparoscopic extraperitoneal, defined "conventional". RESULTS: There were no substantial differences between the two techniques, except for a modest lengthening of the operative time, at least initially. CONCLUSION: The results achieved with the adoption of this technique have shown its feasibility, security and undoubted advantages in terms of mini-invasiveness.

[Laparoscopic radical prostatectomy without ligation of the Santorini's venous plexus]. / Prostatectomia radicale video-laparoscopica senza legatura del plesso venoso del Santorini.

BACKGROUND: The goal of our work is to demonstrate how, in the approach to laparoscopic radical prostatectomy, the ligation of Santorini?s dorsal venous complex is not necessary. The retrospective evaluation of video-laparoscopich radical prostatectomy performed at UOC of Jesi with this technical device, showed an increased blood loss compared to a technique that provides for the ligation of Santorini's plexus and, overall, reduced blood loss compared with the average radical open prostatectomy. MATERIAL AND METHODS: 50 video-laparoscopic extraperitoneal radical prostatectomies were evaluated, which were performed with the same technique and by the same operator. The technique involves the non-ligation of Santorini's venous plexus. Surgical, oncological, functional outcomes were considered and in particular blood loss. RESULTS: The results are consistent with those reported in literature, and without ligation of the Santorini's venous plexus , there has been no increase in blood loss intra-and / or post-operatively, or in the percentage of patients with hemotransfusion. CONCLUSION: The video-laparoscopic radical prostatectomy, although a not yet codified and standardized technique for the treatment of organ-confined prostate cancer, did show oncological and functional results basically l similar to open-sky technique. But compared to this, it has certainly demonstrated a lower blood loss and a reduction of morbidity. The non ligation of Santorini's venous plexus, always necessary during open prostatectomy, in our experience is not necessary in video-laparoscopy.

Diagnostic value of PRND gene expression profiles in astrocytomas: relationship to tumor grades of malignancy.

Doppel (PRND) is a paralogue of the mammalian prion (PRNP) gene. It is abundant in testis and, unlike PRNP, it is expressed at low levels in the adult central nervous system (CNS). Besides, doppel overexpression correlates with some prion-disease pathological features, such as ataxia and death of cerebellar neurons. Recently, ectopic expression of doppel was found in two different tumor types, specifically in glial and haematological cancers. In order to address clinical important issues, PRND mRNA expression was investigated in a panel of 111 astrocytoma tissue samples, histologically classified according to the World Health Organization (WHO) criteria (6 grade I pilocytic astrocytomas, 15 grade II low-grade astrocytomas, 26 grade III anaplastic astrocytomas and 64 grade IV glioblastoma multiforme). Real-time PRND gene expression profiling, after normalisation with GAPDH, revealed large differences between low (WHO I and II) and high grade (III and IV) of malignancy (P<0.001). Extensive differences in PRND gene expression were also found within each grade of malignancy, suggesting that PRND mRNA quantitation might be useful to distinguish astrocytoma subtypes, and important in disease stratification and in the assessment of specific treatment strategies.

[Bilateral and monolateral dolichoarteriopathies (Kinking, Coiling, Tortuosity) of the carotid arteries and atherosclerotic disease. An ultrasonographic study]. / Dolicoarteriopati (Kinking, Coiling, Tortuosità) delle arterie carotidi bilaterali e monolatrali e malattia aterosclerotica. Studio ultrasonografico.

BACKGROUND: The aim of this study, performed in outpatients, is to evaluate, using echo color Doppler examination, atherosclerotic lesions of extracranial carotid arteries in subjects with bilateral, and monolateral dolichoarteriopathies. METHODS: A total of 316 subjects, 77 with bilateral and 239 with monolateral dolichoarteriopathies, were studied. Patients were divided into: 79 subjects without cardiovascular risk factors and 237 subjects with cardiovascular risk factors (98 with arterial hypertension alone, 85 with arterial hypertension associated with other cardiovascular risk factors, 54 with hypercholesterolemia, diabetes, cigarette smoking, alone or associated). Myointimal thickness, as a mean of 5 measurements at common carotid level, 2-3 cm from flow-divider and plaques as a focal thickness =/> 2 mm, were evaluated. RESULTS: In 77 patients with bilateral dolichoarteriopathies, compared to 239 patients with monolateral dolichoarteriopathies, there were no differences in myointimal thickening, and in the prevalence of carotid plaques; no difference was observed between sexes. Both in subjects with and without cardiovascular risk factors myointimal thickness and carotid plaques didn't show any significant differences in bilateral compared with monolateral dolichoarteriopathies. CONCLUSIONS: In conclusion, bilateral, compared with monolateral, dolichoarteriopathies of the carotids, do not seem to be a consequence of atherosclerotic lesions.

A 2D Echo color-Doppler study of the extracranial carotid arteries in borderline arterial isolated systolic hypertension.

BACKGROUND: The aim of this study was to evaluate the prevalence of atherosclerotic carotid lesions in isolated systolic borderline arterial hypertension by 2D echo color-Doppler and effect of night-time pressure fall by ambulatory blood pressure monitoring. METHODS: Outpatients from January 1992 to December 1998 were examined. One hundred and twenty normotensive control subjects and 99 isolated systolic borderline untreated hypertensives were studied, based on blood pressure fall were divided into dippers, with nocturnal systolic and/or diastolic blood pressure fall of >10%, and non dippers. Subjects with ischemic heart disease, valvulopathies, heart failure, renal insufficiency, cerebrovasculopathies, hypercholesterolemia (total cholesterol >200 mg/dl) and diabetes. Normotensives and hypertensives were homogenous for cardiovascular risk factors. A thickness of =/> 0.95 mm, calculated as a mean of 5 measurements of the common carotid artery, 2-3 cm from the carotid bifurcation, was considered a sign of myointimal thickening, and the plaque as a focal thickening of =/> 2 mm, based on echogenic characteristics and site. RESULTS: Compared to normotensives, isolated systolic borderline hypertensives, showed carotid arteries with an intima-media thickening (p=0.002) with one or more plaques (p=0.0001) much more frequently, while normal carotid arteries (p=0.0001) were less frequent. In normotensives, like in hypertensives, the prevalence of vasal lesions was not significantly different in dippers compared with non dippers. Plaques were most often localized at level of the common carotid and lesions were hard. CONCLUSIONS: The conclusions is drawn that isolated systolic hypertension is the sign of major vascular atherosclerotic lesions. The night-time pressure fall does not affect the importance of the lesions.

Tolerance to musculoskeletal allografts with transient lymphocyte chimerism in miniature swine.

BACKGROUND: Although transplantation of musculoskeletal allografts in humans is technically feasible, the adverse effects of long-term immunosuppression subject the patient to high risks for correcting a non-life-threatening condition. Achieving immunologic tolerance to musculoskeletal allografts, without the need for chronic immunosuppression, could expand the clinical application of limb tissue allografting. Tolerance to musculoskeletal allografts has been accomplished previously in miniature swine in our laboratory. Although stable, mixed chimerism has been suggested as the mechanism underlying long-term tolerance in a rat limb model, the mechanism of this tolerance induction has not been established. This report explores the possible relationship between hematopoietic chimerism and tolerance to musculoskeletal allografts in swine. METHODS: Twelve miniature swine underwent vascularized musculoskeletal allograft transplantation from histocompatibility complex (MHC) matched, minor antigen-mismatched donors. Eight animals received a 12-day coprse of cyclosporine, one of which was excluded due to subtherapeutic levels. Four recipients were not immunosuppressed. Serial biopsies to assess graft viability and flow cytometry to assess chimerism were performed. Donor and third-party skin grafts were placed on recipients with surviving allografts greater than 100 days to validate tolerance. RESULTS: Both groups developed early peripheral chimerism, but this chimerism became undetectable by postoperative day 19 in the cyclosporine group and by day 13 in the control group. Animals receiving cyclosporine developed permanent tolerance to their allografts, whereas those not receiving cyclosporine rejected their allografts in 6-9 weeks. Animals demonstrating tolerance to their bone allografts also demonstrated prolonged donor skin graft survival. CONCLUSIONS: Induction of tolerance to musculoskeletal allografts can be achieved in the MHC matched swine. Although hematopoietic chimerism is present in the immediate postoperative period, persistent, long-term chimerism does not seem to be necessary for maintenance of such tolerance.

Mixed chimerism and tolerance without whole body irradiation in a large animal model.

Mixed hematopoietic chimerism may provide a treatment for patients with nonmalignant hematologic diseases, and may tolerize patients to organ allografts without requiring chronic immunosuppression. However, the toxicity of the usual conditioning regimens has limited the clinical applicability of this approach. These regimens generally include some level of whole body irradiation (WBI), which is thought to facilitate engraftment either by making room for donor hematopoietic stem cells or by providing sufficient host immunosuppression to enable donor cells to engraft. Here, we have established mixed chimerism across both minor and major histocompatibility barriers in swine, by using high doses of peripheral blood stem cells in the absence of WBI. After mixed chimerism was established, swine leukocyte antigen-matched (SLA-matched) donor skin grafts were tolerated and maintained for a prolonged period, whereas third-party SLA-matched skin was rejected promptly. Donor-matched kidney allografts were also accepted without additional immunosuppression. Because of its low toxicity, this approach has potential for a wide range of clinical applications. Our data may indicate that niches for engrafting stem cells are filled by mass action and that WBI, which serves to empty some of these niches, can be omitted if the donor inoculum is sufficiently large and if adequate host T-cell depletion is achieved before transplant.

Cytokine-mobilized peripheral blood progenitor cells for allogeneic reconstitution of miniature swine.

BACKGROUND: Because of the relative ease of acquisition, increased yield, and improved engraftment characteristics, mobilized peripheral blood progenitor (stem) cells (PBSCs) have recently become the preferred source for hematopoietic stem cell transplantation. In our laboratory, procurement of a megadose of PBSCs is necessary for on-going studies evaluating non-myelosuppressive transplant regimens for the induction of mixed chimerism and allograft tolerance. To exploit hematopoietic growth factor synergy, we have sought to combine growth factors with proven utility to improve PBSC mobilization and maximize our PBSC procurement through an automated collection procedure. METHODS: Mobilization characteristics of PBSCs were determined in 2-5-month-old miniature swine. Animals received either swine recombinant stem cell factor (pSCF, 100 microg/kg) and swine recombinant interleukin 3 (pIL-3, 100 microg/kg), administered intramuscularly for 8 days, or pSCF, pIL-3, and human recombinant granulocyte-colony stimulating factor (hG-CSF), at 10 microg/kg. Leukapheresis was performed beginning on day 5 of cytokine treatment and continued daily for 3 days. RESULTS: Collection of PBSCs from cytokine-mobilized animals via an automated leukapheresis procedure demonstrated a 10-fold increase in the number of total nucleated cells (TNC) (20-30 x 10(10) TNC) compared to bone marrow harvesting (2-3 x 10(10) total TNC). A more rapid rise in white blood cells (WBCs) was seen after administration of all three cytokines compared to pSCF and pIL-3 alone. An increase in colony-forming unit granulocyte-macrophage frequency measured daily from peripheral blood during cytokine treatment, was seen with the addition of hG-CSF to pSCF/pIL-3 correlating well with the rise in WBCs. Similarly, the addition of hG-CSF demonstrated a notable increase in the median progenitor cell yield from the 3-day leukapheresis procedure. Cytokine-mobilized PBSCs were capable of hematopoietic reconstitution. PBSCs mobilized with pSCF/pIL-3 were infused into an SLA-matched recipient conditioned with cyclophosphamide (50 mg/kg) and total body irradiation 1150 cGy. Neutrophil and platelet engraftment occurred on days 5 and 7, respectively, with minimal evidence of graft-versus-host disease. Complete donor chimerism has been demonstrated 331 days after transplant. CONCLUSIONS: Our preliminary results show that in this well-defined miniature swine model, recombinant swine cytokine combinations (pSCF, pIL-3 with or without hG-CSF) successfully mobilize a high yield of progenitor cells for allogeneic transplantation. Furthermore, these cytokine-mobilized PBSCs demonstrate the potential to reconstitute hematopoiesis and provide long-term engraftment in miniature swine.

Thymic transplantation across an MHC class I barrier in swine.

Thymic tissue transplantation has been performed previously in adult mice to induce donor-specific tolerance across allogeneic and xenogeneic barriers. We have now attempted to extend this technique to a large animal preclinical model and describe here our initial studies of allogeneic thymic transplantation in miniature swine. Two miniature swine were thymectomized before thymic tissue transplantation, and two remained euthymic. Donor thymic tissue was harvested from SLA class I-mismatched juvenile pigs and placed into recipient sternocephalicus muscle, kidney capsule, and omentum. A 12-day course of cyclosporin A was started on the day of transplantation. Allogeneic thymic engraftment could only be achieved in euthymic and not in thymectomized miniature swine using this treatment regimen. Both nonthymectomized animals showed good graft development, with evidence of thymopoiesis, as indicated by positive CD1 and host-type SLA class I immunoperoxidase staining of immature graft-infiltrating cells. Both animals also demonstrated donor-specific T cell hyporesponsiveness, as measured by MLR and cell-mediated lympholysis. The thymic grafts continued to develop despite the appearance of high levels of anti-donor specific cytotoxic IgG Abs. Thus, thymic tissue transplanted across an SLA class I barrier can engraft and support host thymopoiesis in euthymic miniature swine. The presence of the host thymus was required for engraftment. These data support the potential of thymic transplantation as part of a regimen to induce donor-specific tolerance to xenogeneic organ grafts.

Peripheral blood progenitor cell mobilization and leukapheresis in pigs.

BACKGROUND AND PURPOSE: The pig is being investigated as an organ donor for humans. Induction of immunologic tolerance to pig tissues in primates would overcome the major immunologic barriers to xenotransplantation. A proven method of inducing tolerance to allografts is by the induction of mixed hematopoietic chimerism by bone marrow transplantation. We are therefore investigating induction of mixed hematopoietic chimerism in the pig-to-baboon model. METHODS: To obtain large numbers of pig hematopoietic cells, leukapheresis was used to collect blood cell products in miniature swine (n = 5) after progenitor cell mobilization by use of a course of hematopoietic growth factors (cytokines), consisting of porcine interleukin 3, porcine stem cell factor, and human granulocyte colony-stimulating factor. RESULTS: Cytokine therapy and leukapheresis were well tolerated. Cytokine therapy increased the total white blood cell count and allowed large numbers of leukocytes (60 x 10(10)) to be obtained by apheresis, of which approximately 0.1% were granulocyte-erythrocyte-monocyte-megakaryocyte colony-forming units (CFU-GEMMs), which are considered to be representative of hematopoietic progenitors with multi-lineage potential. CONCLUSIONS: The combination of cytokine therapy and leukapheresis enables hematopoietic progenitor cells to be obtained safely from miniature swine.

[Simplification in locating and dissecting the levator muscle of the upper eyelid in surgery for ptosis]. / Simplification du repérage et de la dissection du releveur de la paupière supérieure dans la chirurgie du ptosis.

BACKGROUND: While repairing eyelid ptosis by aponeurotic resection by anterior approach, the risk of damaging the levator complex and the conjunctiva is significant. In order to simplify the dissection between Müller's muscle and the underneath conjunctiva, we use a modification of the usual surgical technique. METHODS: Before the skin incision, the eversion of the upper eyelid allows to dissect the conjunctiva from the Müller's muscle under direct visual control, starting from the upper tarsal margin. A silicone band is then passed through the so created horizontal subconjunctival tunnel. The upper eyelid can be physiologically replaced, and the levator muscle aponeurosis exposed. The two ends of the band are then pulled on surface through two lateral incision performed close to the upper tarsal edge. Now the band plays the role of a useful landmark: every tissue above the band is levator complex; when stretched downwards, it points the upper edge of the tarsal plate. We operated by this technique 24 eyes, affected of acquired or congenital ptosis. Fourteen eyelids had already undergone ptosis surgery elsewhere. RESULTS: We achieved good-to-excellent results in all cases, without any important postoperative complications. CONCLUSIONS: The proposed manoeuvre makes easier the dissection of the inner aspect of the levator complex, because of the material control. Therefore it minimises the tissue trauma and the postoperative complications, particularly in complicated cases characterised by scarring and fibrosis.

Rhegmatogenous retinal detachment with posteriorly located break treated with temporary buckle. A case report.

A rhegmatogenous retinal detachment with a break 19 mm from the limbus in the nasal quadrant was treated with a temporary balloon buckle. This technique was applied to avoid the surgical trauma of vitreous surgery. The operation was successful and the procedure appears to be valid for buckling posterior breaks.