The NATO project: nanoparticle-based countermeasures for microgravity-induced osteoporosis.

Recent advances in nanotechnology applied to medicine and regenerative medicine have an enormous and unexploited potential for future space and terrestrial medical applications. The Nanoparticles and Osteoporosis (NATO) project aimed to develop innovative countermeasures for secondary osteoporosis affecting astronauts after prolonged periods in space microgravity. Calcium- and Strontium-containing hydroxyapatite nanoparticles (nCa-HAP and nSr-HAP, respectively) were previously developed and chemically characterized. This study constitutes the first investigation of the effect of the exogenous addition of nCa-HAP and nSr-HAP on bone remodeling in gravity (1 g), Random Positioning Machine (RPM) and onboard International Space Station (ISS) using human bone marrow mesenchymal stem cells (hBMMSCs). In 1 g conditions, nSr-HAP accelerated and improved the commitment of cells to differentiate towards osteoblasts, as shown by the augmented alkaline phosphatase (ALP) activity and the up-regulation of the expression of bone marker genes, supporting the increased extracellular bone matrix deposition and mineralization. The nSr-HAP treatment exerted a protective effect on the microgravity-induced reduction of ALP activity in RPM samples, and a promoting effect on the deposition of hydroxyapatite crystals in either ISS or 1 g samples. The results indicate the exogenous addition of nSr-HAP could be potentially used to deliver Sr to bone tissue and promote its regeneration, as component of bone substitute synthetic materials and additive for pharmaceutical preparation or food supplementary for systemic distribution.

Colocutaneous fistula through ulcerative colitis and cancer to the pyoderma gangrenosum: a never-ending story for a single patient. Case report.

BACKGROUND: Inflammatory bowel diseases may be associated with many extraintestinal complications, that in some cases can represent the first onset of these disorders. In particular during the course of the disease, Ulcerative Colitis develops extraintestinal manifestations very frequently. One of the rarest is pyoderma gangrenosum, a noninfectious neuthrophilic dermatosis, that can involve most commonly legs but also other parts of the skin or mucosas. It can be idiopathic or associated with gammopathies, vasculitis, chronic arthritis or, like in our case, with inflammatory bowel disease and malignancies. CASE PRESENTATION: A 38-year-old man was referred to our Department with a colo-cutaneous fistula in the left quadrant of abdominal wall. In the anamnesis he reported a trauma during a soccer match three weeks before. Through a CT scan and endoscopy with biopsy an inflammatory bowel disease with a segmental colitis and stenosis was diagnosed. After medical therapy, an initial radiological drainage and a period of parenteral nutrition, he underwent a left hemicolectomy. Despite the previous endoscopic biopsy the histopathological examination put in evidence not only inflammatory disease (in particular Ulcerative Colitis) but also a colorectal tumor pT4pN0. After the full recovery before chemotherapy he has developed on the chest and on the abdomen some painful nodules, with central necrosis, one of those in contact with one of the ribs. Through TC and RM it was impossible to understand the precise nature of these skin lesions. With biopsy a pyoderma gangrenosum was diagnosed and treated until complete resolution. DISCUSSION AND CONCLUSION: Management of inflammatory bowel diseases can be a true challenge, not only for the intestinal manifestations, but also for all the other features not related to gut. In some cases the same patient can develop many complications, such as malignancies or rare cutaneous diseases. Despite the initial surprise for such a weird evolution in a same patient, from fistula to inflammatory disease to cancer and finally to pyoderma gangrenosum, to face every single complication following consolidated diagnostic and pathological paths has been the correct strategy for controlling the disease.

Heterogeneous and self-organizing mineralization of bone matrix promoted by hydroxyapatite nanoparticles.

The mineralization process is crucial to the load-bearing characteristics of the bone extracellular matrix. In this work, we have studied the spatiotemporal dynamics of mineral deposition by human bone marrow mesenchymal stem cells differentiating toward osteoblasts promoted by the presence of exogenous hydroxyapatite nanoparticles. At the molecular level, the added nanoparticles positively modulated the expression of bone-specific markers and enhanced calcified matrix deposition during osteogenic differentiation. The nucleation, growth and spatial arrangement of newly deposited hydroxyapatite nanocrystals have been evaluated using scanning micro X-ray diffraction and scanning micro X-ray fluorescence. As leading results, we have found the emergence of a complex scenario where the spatial organization and temporal evolution of the process exhibit heterogeneous and self-organizing dynamics. At the same time the possibility of controlling the differentiation kinetics, through the addition of synthetic nanoparticles, paves the way to empower the generation of more structured bone scaffolds in tissue engineering and to design new drugs in regenerative medicine.

Wool fibril sponges with perspective biomedical applications.

Sheep's wool was used as a natural source to prepare keratin microfibril sponges for scaffolding, by disruption of the histological structure of the fibres through mild alkali treatment, followed by ultrasonication, casting and salt-leaching. The wool sponges showed highly interconnected porosity (93%) and contain intrinsic sites of cellular recognition that mimic the extracellular matrix (ECM). They displayed good thermal and water stability due to the conversion of disulphide cystine bonds into shorter monosulphide lanthionine intermolecular bonds, but significantly swelled in water, because of the high hydrophilicity and porosity, with a volume increasing up to 38%. Nevertheless, sponges were stable in water without structural changes, with a neutral pH in aqueous media, and showed excellent resilience to repeated compression stresses. According to in vitro biocompatibility assays, wool fibril sponges showed a good cell adhesion and proliferation as proved by MTT, FDA assays and SEM observations. The unique structure of the cortical cell network made by wool keratin proteins with controlled-size macro-porosity suitable for cell guesting, and nutrient feeding, provides an excellent scaffold for future tissue engineering applications.

Zenker diverticulum in the right side of the neck resembling a thyroid mass at ultrasound.

Zenker’s diverticulum represents the most common form of pharyngo-oesophageal diverticula usually occurring on the left side of the neck. Due to its anatomical proximity to the thyroid, it can mimic a thyroid mass. Here we describe the case of an asymptomatic 49-year-old man referred to the Thyroid Clinic of the Policlinico Umberto I Hospital-“Sapienza” University of Rome for thyroid sonography due to a family history of autoimmune thyroid disease. The patient’s thyroid blood tests did not reveal any abnormalities. The sonographic examination showed a dishomogeneus and hypoechoic thyroid gland. In addition, in the third middle of the right lobe, a mass (with a diameter greater than 26 mm), with heterogeneous internal echogenicity, hypoechoic margins and internal hyperechoic spots was recorded, with no appreciable flow at the Doppler evaluation. The TI-RADS score was 4c. Hence, the patient underwent ultrasound-guided fine-needle aspiration cytology that revealed the presence of squamous cells without cytological atypia, erythrocytes, muscular and vegetable fibres, colonies of bacteria in the absence of inflammatory infiltrate. This was consistent with the diagnostic hypothesis of oesophagus diverticulum, which was confirmed by means of a barium-swallow oesophagography. This case report underlines the possibility that a suspicious thyroid mass may result from a Zenker’s diverticulum, even if located on the right side, especially if the lesion has a heterogeneous echo-texture, a hypoechoic rim and internal hyperechoic spots.

Lymph node metastases from differentiated thyroid carcinoma: does radioiodine still play a role?

OBJECTIVES: Ultrasonography and surgery have now become the elective diagnostic and therapeutic tools for neck lymph node (LN) metastases from differentiated thyroid carcinoma (DTC), reserving radioiodine therapy (RAI) for surgery failures. Aim of the present retrospective study was to evaluate results of RAI in cases of LN metastases displaying (131)I uptake over a long-term observation period and its possible role today. MATERIALS AND METHODS: From a series of 1276 patients who had undergone surgery for DTC, 130 cases were selected showing (131)I uptaking LN metastases, detected during follow-up scans and who were then submitted to surgery and/or RAI. Patients were divided into groups according both to extent of surgery, with/without lymphectomy, and to following treatment and outcome. RESULTS: The initial surgical approach does not seem to significantly influence the outcome. (131)I therapy alone, sometimes at low doses, can be very effective in the management of LN metastases detected at Whole Body Scan, but multiple doses are often needed. The age at diagnosis is confirmed as a negative prognostic factor. CONCLUSIONS: Considered radioprotection questions, RAI may solve (131)I uptaking LN metastases, above all if < 10 mm. For larger LN metastases and in the case of failure of RAI surgical excision is mandatory, while a subsequent therapeutic dose of (131)I could be useful to reveal incomplete excision.

Subfascial endoscopic perforator surgery (SEPS) in chronic venous insufficiency. A 14 years experience.

INTRODUCTION: Subfascial Endoscopic Perforator Surgery (SEPS) enables the direct visualization and section of perforating veins. Morbidity and duration of hospitalization are both less than with conventional open surgery (Linton's or Felder's techniques). PATIENTS AND METHODS: A total of 322 legs from 285 patients with a mean age of 56 years (range 23-90) were treated at our Department from May 1996 to January 2010. In 309 cases, an endoscope (ETM Endoskopische Technik GmbH, Berlin, Germany) was introduced through a transverse incision approximately 1.5 cm in length and 10 cm from the tibial tuberosity, as with Linton's technique. A spacemaker balloon dissector for SEPS, involving a second incision 6 cm from the first, was used in only 13 cases. RESULTS: The procedure used in each case was decided on the basis of preoperative evaluation. SEPS and stripping were performed in 238 limbs (73.91%), SEPS and short stripping in 7 limbs (2.17%), SEPS and crossectomy in 51 limbs (15.84%), and SEPS alone in 26 limbs (8.07%). 103 patients presented a total of 158 trophic ulcers; the healing time was between 1 and 3 months, with a healing rate of 82.91% after 1 month and 98.73% after 3 months. CONCLUSION: Subfascial ligature of perforating veins is superior to sclerotherapy and minimally invasive suprafascial treatment for the treatment of CVI. It is easy to execute, minimally invasive and has few complications.

Heterotaxia: radiological and surgical observations in a case of polysplenic syndrome.

We describe a case of asymptomatic polysplenic syndrome as suggested by ultrasonography, gastrointestinal series, computed tomography, magnetic resonance imaging scans and arteriography, and confirmed at surgery. Spleen malformation is frequently associated with heterotaxia and other malformations of the thoracic and abdominal organs, but no pathognomonic features can be detected in relation to this condition. In our patient, imaging studies revealed the presence of a polylobulated spleen in an atypical position below the liver, associated with malpositioning of the gastric fundus and gastro-oesophageal junction with interruption of the inferior vena cava, but no congenital heart disease. In conclusion, polysplenic syndrome is a rare clinical condition, occasionally found in asymptomatic adults. Radiological detection could be mistaken for mediastinal or abdominal pathological masses, and only a thorough study performed with several different radiological methods can determine the precise anatomy of the structures involved so as to be able to plan surgery where necessary.

Nitric oxide prevents inducible cyclooxygenase expression by inhibiting nuclear factor-kappa B and nuclear factor-interleukin-6 activation.

Stimulation of J774 macrophages with lipopolysaccharide (LPS) leads to the release of large amounts of prostaglandins (PGs) generated by the inducible isoform of cyclooxygenase (COX-2). Nitric oxide (NO), a pleiotropic free radical, has been demonstrated to modulate the release of a broad range of inflammatory mediators, amongst these PGs. In the present study we investigated the molecular mechanism by which NO affects cyclooxygenase pathway. Incubation of J774 cells with LPS caused an increase of prostaglandin E2 production and COX-2 protein expression which was prevented in a concentration-dependent fashion by pre-incubating cells with sodium nitroprusside (SNP) and S-nitroso-glutathione (GSNO), two NO-generating agents. Electrophoretic mobility shift assay indicated that both NO-generating agents blocked LPS-induced activation of nuclear factor-kappaB (NF-kappaB) by increasing IkappaB-alpha protein expression and blocking nuclear translocation of NF-kappaB subunits p50 and p65. SNP and GSNO also inhibited nuclear factor-interleukin-6 (NF-IL6) activation. These results show for the first time that SNP and GSNO down-regulate LPS-induced COX-2 expression by inhibiting NF-kappaB and NF-IL6 activation and suggest a negative feed-back mechanism that may be important for limiting excessive or prolonged PGs production in pathological events.

Protective role of nuclear factor kappa B against nitric oxide-induced apoptosis in J774 macrophages.

We investigated the role of constitutive transcription factor nuclear factor kappaB (NF-kappaB) in nitric oxide (NO)-mediated apoptosis in J774 macrophages. Our results show that NF-kappaB is present in untreated J774 cells in a form constitutively active. Incubation of cells with sodium nitroprusside (SNP) and S-nitroso-glutathione (GSNO), two NO-generating compounds, caused: (a) inhibition of constitutive NF-kappaB/DNA binding activity; (b) decrease of cell viability; (c) DNA fragmentation; (d) ApopTag positivity. Pyrrolidine dithiocarbamate (PDTC) and N-alpha-para-tosyl-L-lysine chloromethyl ketone (TLCK), two inhibitors of NF-kappaB activation, showed the same effects of both NO-generating compounds. Furthermore, SNP and GSNO as well as PDTC and TLCK significantly increased the cytoplasmic level of IkappaBalpha. All together these results demonstrate that constitutive NF-kappaB protects J774 macrophages from NO-induced apoptosis. Moreover, these findings show, for the first time, that NO-generating compounds may induce apoptosis in J774 macrophages by down-regulating constitutive NF-kappaB/DNA binding activity and suggest a novel mechanism by which NO induces apoptosis.

[Recurrences after thyroid lobectomy for benign thyroid diseases: analysis of a clinical-instrumental follow up]. / Le recidive dopo lobectomia per tireopatia benigna: analisi di un follow up clinico-strumentale.

The authors propose to verify the therapeutical role of lobectomy in confronting monolobar nodular thyroid disease. In a period of 4 years (1990-1993), forty-four patients (30 women, 14 men) underwent thyroid lobectomy for a benign disease and follow-up treatment for 3 years on a semestral basis. All patient were studied both before the operation and during the follow-up using US and measuring the levels of T3, T4 and TSH; 34 patients (77.2%) had undergone a scintigraphy before surgery. The indication towards surgical intervention was, in 16 patients, consequent to the diagnosis of monolobar uninodular goiter and in 28 patients to monolobar plurinodular goiter. The operation involves thyroidal extracapsular lobectomy with isthmectomy. There were no temporary and/or permanent recurrential lesions, nor there were any significant hypocalcaemic alterations. The US description of the follow-up has been classified in four groups: normal, non-homogeneity (presence of simple parenchymal non-homogeneity), recurrence/non-homogeneity (presence of both pseudonodular forms and parenchymal non-homogeneity), recurrence (presence of recurrence with nodule(s) whose diameter is greater than 5 mm.). From the US follow-up examination, it was shown that 14 out of 44 patients (31.8%) presented alterations in the residual lobe wich were not identifiable in the pre-operative US examination (3 recurrences = 6.81%; recurrence/non-homogeneity = 2.27%; 10 non-homogeneity = 22.7%). Thirteen out of 14 patients are treated with substitutive ormonal therapy Out of these 30 patients whose residual lobe shows no alterations using US, 23 (70.6%) are treated with substitutive ormonal therapy. The authors have underlined a substantial percentage difference between the incidence of normality of the residual lobe during follow-up to lobectomy due to adenoma (normality 88.8%) and to lobectomy due to goiter-related lesions (normality 60%). During US follow-up examinations, abnormality percentages equal or superior to 30%--as those noticed after lobectomy due to goiter-related lesions underline the doubts wich characterize any form of partial thyroidal asportation (4 recurrence in 35 cases of monolobar uni- or plurinodular goiter = 11.4%); also, in the authors' experience, substitutive ormonal therapy has seemingly no effect on the recurrence incidence. According to the authors, the lobectomy can occupy a therapeutical role in thyroid diseases with monolobar expression, but to avoid an elevated number of recurrences, a rigorous evaluation of the real extension of the nodular disease is indispensable.

Longitudinal measurement of amniotic fluid index in term pregnancies and its association with intrapartum fetal distress.

OBJECTIVE: The aim of the present study was to evaluate the dynamic changes in serially obtained amniotic fluid index values and to determine any association with intrapartum fetal distress in a term population. MATERIALS AND METHODS: All patients, > or = 40 weeks of gestational age, evaluated at the Institute of Obstetrics and Gynecology, 'G. Salesi' Hospital, University of Ancona, between January 1, 1994, and December 31, 1995, participated in this longitudinal study. Women with an amniotic fluid index of > 50 mm, who also demonstrated a reactive nonstress test, underwent semiweekly amniotic fluid assessment until spontaneous labor. After 42 gestational weeks, the patients underwent an elective induction of labor. All patients were managed with continuous electronic fetal heart rate monitoring throughout labor. The incidence of intrapartum fetal distress, and meconium staining of amniotic fluid were evaluated with respect to the amniotic fluid index. RESULTS: Of the 117 patients that were evaluated by ultrasound, 83 women had multiple amniotic fluid index measurements and were enrolled in the study. A serial decrease in amniotic fluid index was documented in 54 women; the mean decrease per week was 20.7 +/- 15.4%. An increase in amniotic fluid index was noted in 17, while 11 women showed no change in amniotic fluid index over time. The 14 patients who underwent cesarean section for fetal distress had a significantly lower amniotic fluid index (p < 0.001) at the last sonographic examination than the normal outcome group. Significant differences were also observed for a serial decrease in the amniotic fluid index within a week (p < 0.001). The sensitivity and specificity of the 30% serial decrease in the amniotic fluid index cutoff point, with respect to intrapartum fetal distress were 86 and 93%, respectively. CONCLUSION: Longitudinal measurement of the amniotic fluid index seems to be an effective method in predicting intrapartum fetal distress in a term population.

Prophylactic transabdominal amnioinfusion in oligohydramnios for preterm premature rupture of membranes: increase of amniotic fluid index during latency period.

OBJECTIVE: This study was designed to: (i) evaluate the effect of amnioinfusion on the latency period in patients with oligohydramnios for preterm premature rupture of membranes, and (ii) to investigate the relationship between changes in the amniotic fluid index and fetal heart rate short-term variability by computerized Hewlett-Packard cardiotocography, longitudinally estimated before and after prophylactic amnioinfusion. MATERIALS AND METHODS: All singleton pregnancies with prolonged premature rupture of membranes after 25 weeks of gestation and seen at the Institute of Obstetrics and Gynecology, University of Ancona (Italy), between January 1994 and June 1995 were included in the study. Transabdominal amnioinfusion with 150-350 ml warmed normal saline (25-50 ml/min) was performed at weekly intervals. Amniotic fluid volume was assessed ultrasonographically by means of the four-quadrant technique on a weekly basis before and after each amnioinfusion, as well as the short-term variability by a Hewlett-Packard computerized cardiotocographic system. RESULTS: 18 women were enrolled and underwent prophylactic transabdominal amnioinfusion at weekly intervals until delivery. Eighteen controls, who did not undergo prophylactic amnioinfusion, were recruited from our 1992-1993 series and included in the study. The median interval between premature rupture of membranes and delivery was 3.0 weeks (range 1-8 weeks), with an average delivery age of 33.0 weeks (range 27-36 weeks). The latency period was significantly longer in patients who underwent prophylactic amnioinfusion (mean +/- SD, 4.1 +/- 1.7 weeks) than in controls(1.7 +/- 1.0 weeks; p < 0.001). An increase in both the weekly amniotic fluid index (linear regression analysis r = 0.8, p = 0.03) and the weekly short-term variability (linear regression analysis r = 0.82, p = 0.02) was observed among patients who underwent prophylactic amnioinfusion. A direct relationship was observed between the amniotic fluid index and short-term variability (linear regression analysis r = 0.54, p = 0.04). The mean values of fetal movements recorded by computerized tomography during the 20 min of observation significantly increased after amnioinfusion in comparison with those before it (2.6 +/- 0.9 and 0.9 +/- 0.7 respectively; p = 0.001). CONCLUSION: The present study has shown a positive effect of prophylactic transabdominal amnioinfusion on the latency period in patients with preterm premature rupture of membranes and oligohydramnios. Among the patients who underwent amnioinfusion, an interesting improvement in fetal heart rate short-term variability was associated with the progressive increase in amniotic fluid volume, as an expression of fetal well-being.

[Evaluation of the efficacy of image-guided drainage in the treatment of abdominal fluid collections]. / Valutazione dell'efficacia del drenaggio "imaging guidato" nel trattamento delle raccolte addominali.

In this paper the Authors report their experience in the diagnosis and management of abdominal fluid collections either primary or secondary to surgery. Sixty-eight patients with abdominal fluid collections were considered: in 28 cases an imaging guided percutaneous drainage was performed, while in 40 cases patients were treated with medical or surgical therapy. The Authors describe the different techniques, the approaches and the types of catheter used on the basis of the localization of the collections. The results show the efficacy of drainage procedures in 89% of the patients treated, without any major complication. Some considerations comparing patients treated with percutaneous drainage and patients who underwent different therapy as well as a review of the international literature are also reported. In conclusion the Authors affirm that percutaneous imaging guided drainage is the treatment of choice for abdominal fluid collections anatomically accessible, for the high effectiveness, good tolerability, low cost and minimal incidence of major complications.

[Preoperative diagnosis of thyroid nodule. Ultrasonography-guided fine-needle aspiration]. / In tema di diagnosi di natura preoperatoria del nodulo tiroideo. L'FNA ecoguidata.

Nodular thyroid pathology weighs heavily for 4-5% of the general population, being the most common among the endocrinopathies. Because of this these diagnostic methods have gained more and more importance as FNA able to select already in the pre-operation phase the malignant nodules from the benign ones and to decide on the most suitable surgical intervention. In the period of time between January 1989 and June 1992 in the Institute of the III Surgical Clinic in Rome there have been performed 403 FNA. From the whole total we have extrapolated 221 patients. In this study we stressed how the FNA method has a sensibility of 80%, a specificity of 93% and afterwards an accurate diagnosis of 92%. Besides we have verified that sonography and radionuclide scanning have a truth worthiness respectively of 64% and 52%. The FNA in our opinion allows us to have in the pre-operation phase a much more accurate diagnosis as regards that obtained with only sonography and radionuclide scanning therefore the latter methods must be considered complementary to FNA.

[Role of diagnostic imaging in primary and secondary hyperparathyroidism]. / Il ruolo della diagnostica per immagini nell'iperparatiroidismo primario e secondario.

The authors take in consideration the sensitivity of imaging methods in localization of adenomas and hyperplastic glands in primary and secondary hyperparathyroidism. Advantages and limits of Ultrasound, Computed Tomography, Magnetic Resonance Imaging and Dual Isotope Subtraction Scintigraphy are considered, specifying the correct technique of execution and the indications for each examination. The sensitivity of these imaging methods is fairly good, but inferior in comparison to surgical exploration of the neck; it has been seen that this value improves sensibly when a two-study combination is considered as a single test. After a wide review of the literature and considering their experience, the authors stress the opportunity of a routinary preoperative use of diagnostic imaging, both in primary intervention and in case of reintervention. According to the authors US and MRI are the best imaging modalities: US are of simple execution, non-invasive and have a high sensitivity in the localization of adenomas in the typical location; MRI is a multiplanar technique, has a high contrast resolution and a very good capability of exploring the neck and mediastinum, allowing the localization of ectopic adenomas.

[Ultrasonography and the kidney: a screening method?]. / Ecografia e rene: metodica di screening?

The Authors retrospectively analyzed 733 hepatic U.S. involving non symptomatic urologic patients. The study detected 23 renal cysts, 3 lithiasis (with associated hydronephrosis in two) and 3 kidney neoplasms. It is concluded that U.S. represents a reliable and useful screening method for renal carcinoma, therefore it should be routinely adopted.