FMD empty capsids combined with the Immunostant Particle Adjuvant -ISPA or ISA206 induce protective immunity against foot and mouth disease virus.

Foot and Mouth Disease Virus (FMDV) causes economy losses and is controlled by vaccination in many countries. Vaccine formulations based on empty capsids or Virus-Like Particles (VLPs) have the advantage of avoiding the biological hazard of using infectious FMDV, albeit are poorly immunogenic. Recently, we have described that ISPA a new Immune Stimulating Complex adjuvant, is useful to improve the response against FMD of vaccines that use inactivated virus. Now, the adjuvant effects of ISPA and ISA 206 (water/oil/water) on a VLPs-based FMD vaccine were evaluated. VLPs (strain A/Argentina/2001) were obtained in mammalian cell cultures and their elicitation of an immune response against FMDV with and without ISPA or ISA 206 was evaluated in mice as a first approach. Notably, VLPs-ISPA and VLPs-ISA 206 vaccines induced protection against viral challenge in 100 % of mice, while protection induced by VLPs alone was of 40 %. Total and neutralizing FMDV antibodies were higher in the VLPs-ISPA and VLPs-ISA 206 groups compared to the VLPs group. VLPs-ISPA induced significantly higher (p < 0.001) IgG1, IgG2a, IgG2b and IgG3 titers than the VLPs vaccine. Moreover, in comparison with non-adjuvanted VLPs, VLPs-ISPA and VLPs-ISA 206 elicited an increased virus-specific T response, including higher IFNγ+/CD8 + lymphocyte production in mice. When these vaccines were tested in calves, antibody titers reached an Expected Percentage of Protection (EPP) above 90 % in the case of the VLPs-ISPA and VLPs-ISA 206 vaccines, while, in the VLPs group, EPP reached 25 %. IFNγ levels secreted by mononuclear cells of VLP-ISPA-vaccinated cattle were significantly higher than in the VLPs group. Overall, the results demonstrate that VLPs-ISPA or VLPs-ISA 206 are promising formulations for the development of a novel FMD vaccine.

Design and fabrication of novel polymeric biodegradable stents for small caliber blood vessels by computer-aided wet-spinning.

Biodegradable stents have emerged as one of the most promising approaches in obstructive cardiovascular disease treatment due to their potential in providing mechanical support while it is needed and then leaving behind only the healed natural vessel. The aim of this study was to develop polymeric biodegradable stents for application in small caliber blood vessels. Poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] (PHBHHx), a renewable microbial aliphatic polyester, and poly(ε-caprolactone), a synthetic polyester approved by the US Food and Drug Administration for different biomedical applications, were investigated as suitable polymers for stent development. A novel manufacturing approach based on computer-aided wet-spinning of a polymeric solution was developed to fabricate polymeric stents. By tuning the fabrication parameters, it was possible to develop stents with different morphological characteristics (e.g. pore size and wall thickness). Thermal analysis results suggested that material processing did not cause changes in the molecular structure of the polymers. PHBHHx stents demonstrated great radial elasticity while PCL stents showed higher axial and radial mechanical strength. The developed stents resulted able to sustain proliferation of human umbilical vein endothelial cells within two weeks of in vitro culture and they showed excellent results in terms of thromboresistivity when in contact with human blood.

Influence of FBG sensors length on temperature measures in laser-irradiated pancreas: theoretical and experimental evaluation.

Temperature distribution T(x,y,z,t) in tissue undergoing Laser-induced Interstitial Thermotherapy (LITT) plays a crucial role on treatment outcome. Theoretical and experimental assessment of temperature on ex vivo laser-irradiated pancreas is presented. The aim of this work is to assess the influence of thermometers dimensions on temperature measures during LITT. T(x,y,z,t) inside tissue is monitored by optical sensors, i.e., Fiber Bragg Gratings (FBGs): three FBGs with lengths of 10 mm and nine FBGs of 1 mm, at different distances (2 mm, 5 mm and 10 mm) and different quotes (0 mm, 2 mm and 4 mm) from the laser fiber tip are used. Theoretical punctual T(x,y,z,t) is averaged out on both 10 mm and 1 mm in order to compare numerical predictions with experimental data. Results demonstrate the influence of FBG length on T(x,y,z,t) measures. This phenomenon depends on the distance between sensor and applicator: it is particularly significant close to the applicator tip (2 mm) because of the high spatial T(x,y,z,t) gradient within the tissue. Both theoretical results and experimental ones show that just at a distance of 10 mm from the tip, differences between T(x,y,z,t) provided by FBGs of 10 mm and 1 mm are negligible.

Cancer in pregnancy: maternal and fetal implications on decision-making.

BACKGROUND: Cancer complicates one out of 1,000 pregnancies. No standardized therapeutic interventions have been reported for these patients. METHODS: Fifteen patients with cancer during pregnancy were diagnosed between 6.5 and 36 weeks of gestational age between January 1991 and December 2007. RESULTS: Among the 15 cases one patient with early diagnosis (11 weeks) asked for interruption of pregnancy, two patients rejected chemotherapy in order to avoid fetal effects, seven patients underwent surgery during the first or second trimester, and two patients agreed to start the treatment only after delivery. Standard platinum-based chemotherapy (cisDDP) was postponed in six patients to the second trimester (administered after surgery in 2 cases). Chemotherapy was started between 18.3 and 29.6 weeks (median 22.3 weeks). One patient had pPROM (22.3 weeks) after chemotherapy with cisDDP. Ten patients were delivered by elective cesarean section and three by vaginal delivery. Mean gestational age at delivery was 33.5 weeks (range 32.1-40.0); mean weight at birth was 2,550 g (range 1,250-3,450). None of the newborns showed congenital malformations, and all had normal Apgar scores. Anemia occurred in two newborns. At a median follow-up of 56 months (range 2-198 months) all children were well and healthy. Eleven out of 15 mothers are alive and well, and one is alive with disease. An advanced neoplasm was diagnosed in three patients who died. CONCLUSION: When platinum-based chemotherapy is administered during the 2nd-3rd trimester, adverse effects in newborns are comparable to those in the general population. Deliberate treatment delay to achieve fetal viability or to improve fetal outcome may be reasonable for patients with early-stage cancer.

Penetrating brain injuries from a dog bite in an infant.

Brain lesions due to dog bites are not frequent and mainly concern infants in the first years of their life because they are short in height, the size of the infant's head is relatively large compared to the body and the skull bones are thin. We report the case of an infant with bilateral skin, skull and brain injuries secondary to a dog bite, and review the relative reports in the literature. We stress the need to consider the possibility of penetration into the intracranial compartment occurring in cases of dog bites of the scalp, because these wounds on the scalp and the skull may appear relatively limited, small in size and slight, in spite of associated potentially dangerous deeper lesions involving the intracranial structures that may be missed on the first observation. Infection is the main possible complication, and may be the cause of permanent and serious neurological deficits. Prompt diagnosis and therapy are mandatory to avoid complications and to achieve good clinical results.

Angiogenesis in intracranial meningiomas: immunohistochemical and molecular study.

Much of the morbidity of intracranial meningiomas is related to the degree of tumour vascularity and the extent of peritumoural vasogenic oedema. Several studies have shown that vascular endothelial growth factor (VEGF) is up-regulated in meningiomas, although its relationship with tumour vasculature is still unclear. In order to better understand the angiogenic assessment of intracranial meningiomas, we analysed its vascular pattern, both as number and as morphologic configuration of microvessels. Moreover, we investigated the mRNA-VEGF expression, relating this expression to vascular pattern. A total of 40 intracranial meningiomas, classified as benign (31 cases), atypical (7 cases), and anaplastic (2 cases) were analysed. RT-PCR analyses of mRNA-VEGF and competitive-PCR were performed. VEGF expression and microvessel density (MVD) were also immunohistochemically investigated. Grade II-III meningiomas showed numerous small microvessels (mean: 34), while the majority of Grade I showed few larger vessels (mean: 13.09) (P = 0.000003). A microvessel pattern overlapping into atypical subtype was found in eignt of the 31 (25.8%) Grade I meningiomas. A significant association was found between grading and vascular pattern (P = 0.0002), as well as between the MVD and the immunohistochemical expression of VEGF (P = 0.0005). The expression of mRNA agreed with the immunohistochemical expression of the protein (P < 0.0001). A total of 39 cases expressed the 121 VEGF isoform and, among these, 28 cases also expressed the 165 isoform. Only 9 cases expressed both isoforms 165 and 189. Grade II and III meningiomas showed a preponderant expression of soluble isoforms (121 and 165). These results prompt us to speculate that the microvessel pattern could underlie a higher metabolic demand, probably due to a rapid growth with a consequent worse clinical behaviour of the tumour. In this sense, the vascular pattern may be used as a prognostic factor, in order to mostly focus attention on those Grade I meningiomas which have a higher likelihood of either recurrence or development of perilesional oedema. The pattern of vasculature itself seems to be dependent on the types of VEGF isoforms: the Grade II-III meningiomas (that presented numerous microvessels) expressed the soluble isoforms 121 and 165, while the isoform 189 was more frequently detected in Grade I meningiomas.

Telomerase in intracranial meningiomas.

Telomere length maintenance is essential for tumorigenesis; most human tumors stabilize their chromosome ends via the activity of a specialized reverse transcriptase, telomerase, that uses the template region of the RNA moiety complementary to the TTAGGG repeat to synthesize one strand of telomeric DNA. Meningiomas are estimated to constitute between 13% and 26% of primary intracranial tumors. The aim of this study was to evaluate telomerase activity and its messenger expression in meningiomas in relation to their different histologic pattern and grade of cytonuclear atypies, which are associated with relapse, and consequently represent the most important parameter for the evaluation of the clinical behavior of this tumor. Telomerase activity was examined by the telomeric repeat amplification protocol (TRAP) assay in 32 meningiomas (26 typical and 6 atypical/anaplastic). Telomerase messenger expression (hTERT mRNA) was evaluated by reverse transcription-PCR analysis in the same group of tumors. Telomerase activity ranged from undetectable to low levels in 19/26 (73%) of typical meningiomas, while all the atypical/anaplastic meningiomas showed medium-high levels of activity (>3 TPG units, median value), (chi(2) test; p=0.001). The levels of telomerase in terms of its messenger level expression overlapped the activity; a significant association between telomerase activity and hTERT mRNA expression was also found (chi(2) test; p=0.01). Moreover, 2 atypical/anaplastic meningiomas of our series relapsed; in these samples we found high levels of telomerase, both in terms of activity and mRNA expression. Telomerase activity and its hTERT mRNA expression tended to increase as the histologic grading of intracranial tumors increased, suggesting a role of telomerase reactivation in the progression of these tumors. Moreover, our results indicate RT-PCR assay as a rapid tool to identify and quantify telomerase RNA in intracranial meningiomas as in other human tumor models.

Non-pharmacological approaches to chronic headaches: transcutaneous electrical nerve stimulation, lasertherapy and acupuncture in transformed migraine treatment.

In an open, randomized trial, we evaluated transcutaneous electrical nerve stimulation (TENS), infrared lasertherapy and acupuncture in the treatment of transformed migraine, over a 4-month period free of prophylactic drugs. Sixty women suffering from transformed migraine were assigned, after a one month run-in period, to three different treatments: TENS (Group T; n=20), infrared lasertherapy (Group L; n=20) or acupuncture (Group A; n=20). In each group the patients underwent ten sessions of treatment and monthly control visits. In Group T patients were treated for two weeks (5 days/week) simultaneously with three TENS units with different stimulation parameters (I: pulse rate = 80 Hz, pulse width = 120 micros; II: 120 Hz, 90 micros; III: 4 Hz, 200 micros). In Group L an infrared diode laser (27 mW, 904 nm) was applied every other day on tender scalp spots. In Group A acupuncture was carried out twice a week in the first two weeks and weekly in the next 6 weeks. A basic formula (LR3, SP6, LI4, GB20, GV20 and Ex-HN5) was always employed; additional points were selected according to each patient's symptomatology. The number of days with headache per month significantly decreased during treatment in all groups. The response in the groups differed over time, probably due to the different timing of applications of the three methods. TENS, lasertherapy and acupuncture proved to be effective in reducing the frequency of headache attacks. Acupuncture showed the best effectiveness over time.

Meningioma-associated brain oedema: the role of angiogenic factors and pial blood supply.

BACKGROUND: Approximately 60% of meningiomas are associated with perilesional brain oedema. Several aspects have been evaluated in order to understand the pathophysiological mechanisms of oedema (age, sex of the patient, size and location of the tumour, histotype, grading), although at present they have yet to be completely clarified. We focused on pial blood supply, microvascular density (MVD) and angiogenic growth factors (i.e. vascular endothelial growth factor--VEGF) in order to evaluate their putative role in the development of brain oedema. METHODS: We retrospectively studied 55 patients with intracranial meningiomas. Computerized tomography (CT) and angiographic studies were obtained in all cases. The angiograms provided an accurate differentiation between pial and dural blood supply, concomitantly with its semi-quantitative evaluation. The location and the volume of oedema, in relation to the meningioma surface, was evaluated using CT scans, as an oedema index (E/I). We also determined the expression of VEGF and MVD using standard immunohistochemical methods. RESULTS: Thirty-two out of 55 meningiomas presented peritumoural oedema, with an angiographic blush ranging from 2 to 4; VEGF protein was expressed in 27 out of 32 cases, independent of grade or histotype of tumours. In all patients, MVD ranged from 4 to 33.3 vessels (median value: 10.6). A significant relationship was found between the expression of VEGF and MVD (p = 0.0003) and between VEGF and E/I (p = 0.0023). Moreover, the E/I ratio was related to the blush (p = 0.0005). A significant association was also present between VEGF expression and pial blush (p = 0.0001). CONCLUSION: Our data confirm the central role of VEGF and pial blood supply in the pathogenesis of peritumoural oedema and support the hypothesis that the development of oedema in meningioma is vasogenic in type.

Effects of left ventricular pacing on cardiac performance and on quality of life in patients with drug refractory heart failure.

We assessed the effects of left ventricular pacing on echocardiographic and clinical parameters in 13 consecutive patients with heart failure and bundle branch block by means of a controlled acute and medium-term evaluation. Left ventricular pacing induced a significant improvement in left ventricular ejection fraction, Minnesota Living with Heart Failure Questionnaire score, New York Heart Association class, and 6-minute walking test compared with sinus rhythm or right ventricular pacing.

Long-term prospective assessment of left ventricular thrombus in anterior wall acute myocardial infarction and implications for a rational approach to embolic risk.

To prospectively assess the predictive value of left ventricular (LV) thrombus anatomy for defining the embolic risk after acute myocardial infarction (AMI), 2 comparable groups of patients with a first anterior AMI (group A, 97 thrombolysed patients; group B, 125 patients untreated with antithrombotic drugs [total 222]) underwent prospective serial echocardiography (follow-up 39 +/- 13 months) at different time periods. LV thrombi were detected in 26 patients in group A (27%) and in 71 in group B (57%; p <0.005). Embolism occurred in 12 patients (5.4%; 1 in group A [1%] vs 11% in group B [9%], p < 0.04). At multivariate analysis, thrombus morphologic changes were the most powerful predictor of embolism (p <0.001), followed by protruding shape (p <0.01) and mobility (p <0.02). In patients untreated with thrombolysis, a higher occurrence of thrombus morphologic changes (48% vs 8%, p <0.002) and protruding shape (69% vs 31%, p <0.002) were observed, whereas thrombus mobility was similar in the 2 groups (18% vs 8%, p = NS). Thrombus resolution occurred more frequently in thrombolysed patients (85% vs 56%, p <0.002). Thus, after anterior AMI, changes in LV thrombus anatomy frequently occur and appear the most powerful predictor of embolization. A minor prevalence of thrombus, a more favorable thrombus anatomy, and a higher resolution rate may contribute to reduce embolic risk after thrombolysis.

Left atrial appendage function assessed by transesophageal echocardiography before and on the day after elective cardioversion for nonvalvular atrial fibrillation.

We investigated left atrial appendage function by transesophageal echocardiography, on the day after external electrical cardioversion to sinus rhythm, in 41 patients with nonvalvular atrial fibrillation. After cardioversion, appendage contraction synchronized with the electrical and mechanical activity of the atrium, which was restored in about 70% of the patients.

Creative synthesis of visual images is not associated with individual differences.

50 undergraduates were tested on a mental synthesis task aimed at producing creative visual patterns and were administered three questionnaires measuring imagery vividness and control. Analysis did not support a relationship between scores on visual synthesis and imagery and showed that neither kind of score was influenced by sex and studies attended.

Hearing threshold assessment with auditory brainstem response (ABR) and ElectroCochleoGraphy (ECochG) in uncooperative children.

Two-hundred-and-sixty uncooperative children (442 ears) performed auditory brainstem response (ABR) and Electrocochleography (ECochG) in the same diagnostic session under general anaesthesia, and the results obtained with the two different methods were compared. A difference > or = 20 dB between the two methods was found in 134 ears (30.3%). The presence of middle ear effusion and symptoms of a possible central nervous system pathology were considered in order to verify the evidence of a correlation between the difference in ABR-ECochG results and these clinical parameters. The presence of middle ear effusion was not significantly correlated with differences > or = 20 dB (p = 0.1347). On the contrary, the presence of symptoms indicative of a possible central nervous system (CNS) involvement was significantly correlated with differences > or = 20 dB (p = 0.0000). ABR has to be considered the first choice in hearing assessment strategy, either for screening or diagnosis. However, the diagnosis of hearing loss only on the basis of the presence or absence of wave V requires some care in case of suspected central auditory pathway lesions. In these cases, ECochG may be the only reliable diagnostic tool for hearing assessment in uncooperative subjects.

[From the anatomical desk to virtual anatomy]. / Dal tavolo anatomico all'anatomia per immagini.

Teaching of gross Anatomy, the oldest between medical sciences, today suffers the lack of cadavers for notomization, therefore this subject is more theoretical than practical in medical school. The computer techniques could be very useful in this field. Is it possible nowadays to get a software of Virtual Anatomy? The answer is yes. We present in this work a review of the state-of-art of these techniques mainly based on data acquired by computer tomography (CT) or magnetic resonance imaging (MRI) from human. Serial slices obtained from imaging (CT or MRI) can be reconstructed using computers in order to generate a realistic view of the surface of an anatomical object.

Chronic aneurysmatic dilatation: a possible source of lethal embolization in patients with acute myocardial infarction undergoing thrombolysis.

Left ventricular thrombosis is relatively common after acute myocardial infarction, especially in the anterior site, and represents a possible cause of potentially lethal peripheral embolization 1. Therefore, several studies have been performed in order to assess the efficacy of different antithrombotic drugs in resolving the detected thrombi or reducing their embolic potential. Fibrinolytic agents appear effective in this regard: in the majority of cases, they produce complete lysis and resolution of the thrombi. However, this treatment may itself cause embolic complications by producing a rapid fragmentation of thrombus and the subsequent emission of disrupted portions of the intracardiac mass into the systemic vascular bed [2]. This dramatic effect of thrombolysis has suggested the possibility that even the standard treatment of acute myocardial infarction with fibrinolysis implies a danger of embolization in those patients in whom a left ventricular thrombus may be present either from a previous myocardial infarction or from a very early thrombus development. However, this hypothesis has not yet been confirmed by direct observation. We report the case of a patient with a first acute anterior myocardial infarction, in whom the thrombolytic treatment induced lysis and embolization from a left ventricular thrombus present in an aneurysmatic dilatation of the infero-posterior wall due to a previous inferior myocardial infarction.

Combined preoperative chemoradiotherapy followed by radical surgery in locally advanced vulvar carcinoma. A pilot study.

BACKGROUND: Although for decades exenterative surgery has represented the standard treatment for patients with locally advanced vulvar cancer, combined approaches, including preoperative radiation with or without chemotherapy, are now considered the treatment of choice. We report the results of a pilot study on concurrent chemoradiotheraphy followed by radical surgery for patients with locally advanced squamous cell carcinomas of the vulva. METHODS: Thirty-one patients with squamous cell carcinoma of the vulva were treated with two courses of combination chemotherapy mitomycin C, 15 mg/m2 intravenously (i.v.) on Day 1, and 5-fluorouracil, 750 mg/m2 i.v., in continuous 24-hour infusion on Days 1 to 5. Inguinal and pelvic lymph node chains and the vulva were irradiated (starting on the same day as the chemotherapy) up to a total dose of 36 Gy. After a 2-week interval, a second course of chemoradiotherapy was given (18 Gy on the vulvar region only). After 2 weeks, patients underwent radical surgery. RESULTS: An objective response was observed in 22 of 24 primary cases (91.6%) and in 7 of 7 recurrent cases. All but two unresponsive patients underwent radical surgery. The postoperative morbidity rate was 65% (19 of 29 patients), and the mortality rate was 13.8% (4 of 29 patients). Five of nine patients (55%) with biopsy-proven inguinal lymph node metastases showed no residual lymph node disease in the surgical specimen. The recurrence rate was 31.8% and the medial follow-up time was 34 months. CONCLUSIONS: Chemoradiotherapy seems to be effective for squamous cell carcinoma of the vulva. If treatment-related morbidity could be decreased, such a combined approach might offer a new perspectives for a conservative treatment of locally advanced vulvar cancer.

Second surgical treatment of retroperitoneal persistent disease in epithelial ovarian cancer.

AIMS AND BACKGROUND: This report retrospectively analyzes 9 cases of epithelial ovarian cancer with persistent retroperitoneal metastasis after intraperitoneal surgery (without systematic lymphadenectomy) and chemotherapy. METHODS: All 9 patients were diagnosed as FIGO stage I to IV at the time of primary surgery. They received combined postoperative chemotherapy (8 cases with a cisplatin-based regimen and 1 with adriamycin and endoxan). They were submitted to pelvic and paraaortic lymphadenectomy at the National Cancer Institute of Milan during the period 1990-1994. RESULTS: All patients presented no evidence of disease in the abdominal cavity but retroperitoneal metastasis, which was the unique metastatic site. Chemotherapy was administered as adjuvant therapy after lymphadenectomy. Six patients were free of disease for 14 to 61 months. One patient with vaginal recurrence at the 18th month was treated with radiotherapy and chemotherapy, but died of widespread disease 25 months after lymphadenectomy. Two patients with massive positive lymph nodes dies of brain and lung metastasis 20 and 6 months later, respectively. CONCLUSIONS: We conclude that retroperitoneal metastasis may be the only site of persistent disease and that systematic lymphadenectomy technically feasible in this situation to increase the opportunity for local disease control and to obtain a good result.