Cristiano Ialongo

Diabetes as main risk factor in head and neck reconstructive surgery with free flaps.

The aim of our study was to demonstrate the role of certain risk factors in reconstructive head and neck surgery with free flaps. The data taken from the charts of all patients who received free flap for head and neck reconstruction in our department between January 2001 and December 2004 were analyzed. We evaluated the association of preexisting risk factors with the onset of surgical complications such as orocutaneous fistulae, flap infections, hematomas, thrombosis, and necrosis. One hundred and twenty-two free flaps have been used for the reconstruction of head and neck area in 118 patients. Preoperative risk factors included smoking habit (77 patients), alcohol use (6 patients), hypertension (9 patients), diabetes mellitus (8 patients), family history positive for vascular disorders (27 patients), and hypercholesterolemia/hypertriglyceridemia (5 patients). The percentage of full flap survival was 95.08%. Statistical analysis showed that diabetes mellitus (P < 0.01) is significantly associated with a negative prognosis for free flap reconstructive operation, whereas a smoking habit seemed to be at the verge of statistical significance. Therefore, our current practice is to prefer as much as possible the use of local flaps as opposed to free flaps in the reconstruction of head and neck defects in diabetic patients.

A liquid chromatography-tandem mass spectrometry method for the determination of 5-Fluorouracil degradation rate by intact peripheral blood mononuclear cells.

5-Fluorouracil (5-FU) is a major chemotherapy drug used for the treatment of tumors. It is catabolized mainly by dihydropyrimidine dehydrogenase, and patients with a complete or partial deficiency of dihydropyrimidine dehydrogenase activity are at risk of developing severe 5-FU-associated toxicity. The aim of this study was to demonstrate that intact peripheral blood mononuclear cells (PBMCs) can be an effective model to evaluate the degradation rate of 5-FU. We developed a sensitive and specific liquid chromatography-tandem mass spectrometry method to measure in vitro the rate of 5-FU degradation by intact PBMC. 5-FU degradation rate was determined by measuring the decrease of a fixed amount of 5-FU (10 μg/mL) added to a solution of PBMC, after 2 hours incubation, expressed as nanogram per milliliter of 5-FU degraded per minute × 106 cells. Freshly prepared intact PBMC can degrade efficiently in vitro-added 5-FU. The assay consists of 3 steps: (1) PBMC isolation from peripheral blood, (2) PBMC incubation with 5-FU in vitro for different times, and (3) determination of 5-FU amount to calculate the degradation rate. 5-FU was analyzed by a Q Trap 2000 triple quadrupole/ion trap mass spectrometer in the multiple-reaction-monitoring modes. The chromatographic separation was accomplished using a C18 column with a run time of 16 minutes. By analyzing samples from 39 patients with no 5-FU toxicity, the mean 5-FU degradation rate was 1.85 ± 0.50 ng·mL−1·min−1 × 106 cells. The assessment of a test to measure 5-FU degradation rate in PBMC of patients before 5-FU administration could represent a prescreening method for evaluating the possible toxicity of this drug as an aid to set up a personalized medicine approach for each patient.

Understanding the effect size and its measures

The evidence based medicine paradigm demands scientific reliability, but modern research seems to overlook it sometimes. The power analysis represents a way to show the meaningfulness of findings, regardless to the emphasized aspect of statistical significance. Within this statistical framework, the estimation of the effect size represents a means to show the relevance of the evidences produced through research. In this regard, this paper presents and discusses the main procedures to estimate the size of an effect with respect to the specific statistical test used for hypothesis testing. Thus, this work can be seen as an introduction and a guide for the reader interested in the use of effect size estimation for its scientific endeavour.

Iliac crest flap: donor site morbidity.

Introduction:Starting from the 1980s, with the advent of microsurgery, microvascular flaps are used for the reconstruction of wide and complex bone defects of the maxillomandibular district. Compared with conventional and implant-supported prostheses, the free flaps allow aesthetic-functional rehabilitations more adapt to answer to problems that these wide disablements involve. The anatomic characteristics of the crest flap make it one of the best available flap in the maxillomandibular bone reconstruction. Methods:The authors introduce a retrospective analysis of their own experience in the reconstruction of wide and complex bone defects of the maxillomandibular district. Specifically, the attention is focused on the use of the iliac crest flap. The surgical technique of flap preparation is discussed. Moreover, a review of results from international studies about the morbidity of the donor site is presented and compared with own experience. Result:As reported in the literature, the iliac crest flap donor site may encounter several complications. Among these, chronic pain, loss of regional sensibility or paresthesias, hematoma, seroma, walking troubles, unaesthetic scars, abdominal hernia, and loss of the normal bone profile of the hip. Discussion:At present, the use of the iliac crest free flap in the microvascular reconstruction of the complex deficits of the maxillomandibular district represents a well-established method in the experience of the maxillofacial surgeon. Several information about results obtained in the maxillomandibular rehabilitation are available from the literature; however, little attention has been addressed to complications and morbidity of the donor site. Such aspect will be discussed in this work.

Quantitative profiling of oxylipins through comprehensive LC-MS/MS analysis of Fusarium verticillioides and maize kernels

Fusarium verticillioides is one of the most important fungal pathogens causing ear and stalk rot in maize, even if frequently asymptomatic, producing a harmful series of compounds named fumonisins. Plant and fungal oxylipins play a crucial role in determining the outcome of the interaction between the pathogen and its host. Moreover, oxylipins result as signals able to modulate the secondary metabolism in fungi. In keeping with this, a novel, quantitative LC-MS/MS method was designed to quantify up to 17 different oxylipins produced by F. verticillioides and maize kernels. By applying this method, we were able to quantify oxylipin production in vitro – F. verticillioides grown into Czapek–Dox/yeast extract medium amended with 0.2% w/v of cracked maize – and in vivo, i.e. during its growth on detached mature maize ears. This study pinpoints the role of oxylipins in a plant pathogen such as F. verticillioides and sets up a novel tool aimed at understanding the role oxylipins play in mycotoxigenic pathogens during their interactions with respective hosts.

Phlebotomy, a bridge between laboratory and patient

The evidence-based paradigm has changed and evolved medical practice. Phlebotomy, which dates back to the age of ancient Greece, has gained experience through the evolution of medicine becoming a fundamental diagnostic tool. Nowadays it connects the patient with the clinical laboratory dimension building up a bridge. However, more often there is a gap between laboratory and phlebotomist that causes misunderstandings and burdens on patient safety. Therefore, the scope of this review is delivering a view of modern phlebotomy to “bridge” patient and laboratory. In this regard the paper describes devices, tools and procedures in the light of the most recent scientific findings, also discussing their impact on both quality of blood testing and patient safety. It also addresses the issues concerning medical aspect of venipuncture, like the practical approach to the superficial veins anatomy, as well as the management of the patient’s compliance with the blood draw. Thereby, the clinical, technical and practical issues are treated with the same relevance throughout the entire paper.

A clinical and biochemical analysis in the differential diagnosis of idiopathic normal pressure hydrocephalus

Introduction Idiopathic normal pressure hydrocephalus (iNPH) can be misdiagnosed with other neurodegenerative diseases, especially in the early disease stages. Considering the opportunity of the shunt surgery, iNPH should be diagnosed with accuracy. Here, we evaluate the utility of CSF biomarkers and their relationship with clinical features in the diagnosis of iNPH. Methods We performed a multivariate analysis of the CSF levels of Aβ42, t-tau, and p-tau collected from four groups of patients: 14 iNPH, 14 progressive supranuclear palsy (PSP), 14 Alzheimer’s disease (AD), 14 controls (CTL). Diagnostic accuracy of biomarkers was determined by the receiver operating characteristic curve analysis. Statistical correlation was calculated between each CSF biomarker and single clinical items of iNPH. Results Aβ42 levels in iNPH were lower than controls, although not as low as in AD. Likewise, CSF t-tau and p-tau were lower in iNPH than in controls. Of interest, t-tau and p-tau were higher in AD than in controls and hence both t-tau and p-tau were significantly lower in iNPH than in AD. No differences were found between iNPH and PSP. CSF biomarkers levels did not correlate to clinical features of iNPH, whereas two significant correlations emerged within clinical parameters: cognitive impairment was related to gait difficulties, while ventricular enlargement correlated with continence disturbances. Conclusion Measurement of CSF biomarker levels may be helpful in the differential diagnosis between iNPH and AD but not between iNPH and PSP. Both Aβ42 and tau levels appear unrelated to main clinical features of iNPH.

Impact of preanalytical handling and timing for peripheral blood mononuclear cells isolation and RNA studies: the experience of the Interinstitutional Multidisciplinary BioBank (BioBIM).

Multicenter studies and biobanking projects require blood transportation from the participating center to a central collection or diagnostic laboratory. The impact of time delays between venous blood collection and peripheral blood mononuclear cells (PBMC) isolation prior to RNA extraction may affect the quality and quantity of isolated nucleic acids for genomic applications. Thus, standard operating procedure (SOP) optimization for the treatment of biological samples before RNA extraction is crucial in a biological repository. In order to define SOPs for whole blood preservation prior to RNA extraction, we sought to determine whether different blood storage times (0, 3, 6, 10, 24, and 30 hours) prior to PBMCs isolation and storage at –80°C, could affect the quality and quantity of extracted RNA. After spectrophotometric quantification, the quality and integrity of RNA were assessed by agarose gel electrophoresis, RNA integrity number and real time-PCR (RT-PCR). Across the different time points we did not observe significant differences within the first 24 hours of blood storage at room temperature, while a significant loss in RNA yield and integrity was detected between 24 and 30 hours. We conclude that time delays before PBMCs isolation prior to RNA extraction may have a significant impact on downstream molecular biological applications.

Total Automation for the Core Laboratory Improving the Turnaround Time Helps to Reduce the Volume of Ordered STAT Tests

The transition to total automation represents the greatest leap for a clinical laboratory, characterized by a totally new philosophy of process management. We have investigated the impact of total automation on core laboratory efficiency and its effects on the clinical services related to STAT tests. For this purpose, a 47-month retrospective study based on the analysis of 44,212 records of STAT cardiac troponin I (CTNI) tests was performed. The core laboratory reached a new efficiency level 3 months after the implementation of total automation. Median turnaround time (TAT) was reduced by 14.9±1.5 min for the emergency department (p < 0.01), reaching 41.6±1.2 min. In non–emergency departments, median TAT was reduced by 19.8±2.2 min (p < 0.01), reaching 52±1.3 min. There was no change in the volume of ordered STAT CTNI tests by the emergency department (p = 0.811), whereas for non–emergency departments there was a reduction of 115.7±50 monthly requests on average (p = 0.026). The volume of ordered tests decreased only in time frames of the regular shift following the morning round. Thus, total automation significantly improves the core laboratory efficiency in terms of TAT. As a consequence, the volume of STAT tests ordered by hospital departments (except for the emergency department) decreased due to reduced duplicated requests.

Is SOD2 Ala16Val Polymorphism Associated with Migraine with Aura Phenotype

Several studies suggest a role of oxidative stress in the physiopathology of migraine, particularly in the form with aura. In a case-control study, we investigated the association between migraine and superoxide dismutase 1 (SOD1) and superoxide dismutase 2 (SOD2) genes in a cohort of 490 consecutive unrelated Caucasian migraineurs (migraine with aura [MwA], n=107; migraine without aura [MwoA], n=246; chronic migraine [CM], n=137) and 246 healthy controls recruited at our Headache and Pain Unit and stored in the Interinstitutional Multidisciplinary BioBank (BioBIM). Migraine phenotype was carefully detailed using face-to-face interviews. We examined polymorphisms of SOD1 gene (A/C substitution-rs2234694) and SOD2 gene (C/T transition-rs4880-Ala16Val). The rs4880 TT (Val/Val) genotype was associated (p=0.042) with the presence of unilateral cranial autonomic symptoms (UAs) in MwA patients. We also found a mild correlation between SOD2 rs4880 genotype and the type of acute migraine treatment (p=0.048) in MwA patients. Our findings suggest that SOD2 is a disease-modifier gene influencing oxidative mechanisms in MwA. These observations lead to the hypothesis that SOD2 polymorphism may cause a defective control of the oxidative phenomena linked to cortical spreading depression, the neurophysiological hallmark of migraine aura, causing an overstimulation of trigeminal neurons and UAs triggering.