Effect of growth factors and steroid hormones on heme oxygenase and cyclin D1 expression in primary astroglial cell cultures.

Astrocyte activity may be modulated by steroid hormones and GFs. This study investigates the interaction between glucocorticoids or estrogens and GFs on the expression of heme oxygenase-1 (HO-1) and cyclin D1 in astrocyte cultures at 14 days treated for 48 or 60 hr with dexamethasone (DEX) or 48 hr with 17ß-estradiol (E2) alone or with GFs added only in the last 12 or 24 hr. Twelve- or twenty-four-hour epidermal growth factor (EGF) treatment significantly enhanced HO-1 expression in astrocyte cultures pretreated for 48 hr with DEX. A highly significant increase in HO-1 expression was obtained after the last-12-hr EGF treatment in 48-hr E2-pretreated astrocyte cultures; this enhancement was particularly significant in 48-hr E2-pretreated cultures as well as in the last-12-hr insulin-treated ones pretreated for 48 hr with E2. Sixty-hour DEX-alone pretreatment as well as the last-12-hr EGF treatment in 60-hr DEX-pretreated astrocyte cultures showed a significant increase of cyclin D1 expression. A significant decrease of cyclin D1 expression in the last-12-hr insulin-like growth factor-I (IGF-1)-treated cultures pretreated for 60 hr with DEX was observed. A highly significant enhancement in cyclin D1 expression in 14 days in vitro astrocyte cultures pretreated with E2 alone for 48 hr and treated in the last 12 hr with IGF-1 in 48-hr E2-pretreated cultures was found. Finally, the data highlight an interactive dialogue between the growth factors and glucocorticoids or estrogens during the maturation of astroglial cells in culture that may control the HO-1 and cyclin D1 expression as well as proliferating astroglial cells during the cell cycle.

Effect of lipoic acid and α-glyceryl-phosphoryl-choline on astroglial cell proliferation and differentiation in primary culture.

Lipoic acid plays a crucial role as antioxidant and metabolic component of enzymes involved in glucose metabolism of different cell types. Choline alphoscerate (α-glyceryl-phosphoryl-choline [αGPC]) is a semisynthetic derivative of phosphatidylcholines representing, among acetilcholine precursors, a cholinergic drug. In the present study, we evaluated the expression of some proliferation and differentiation markers in 15 or 21 DIV astrocyte cultures treated with 50 µM (+)lipoic acid or (+/-)lipoic acid and/or 10 mM αGPC for 24 hr. In addition, we evaluated the possible genoprotective effect by analysis of DNA status detected by alkaline comet assay. The addition of single drugs [(+)lipoic acid, (+/-)lipoic acid, or αGPC] induced an "upward modulation" of the expression of biomarkers used in our study. On the contrary, the cotreatment with either (+)lipoic acid + αGPC or (+/-)lipoic + αGPC surprisingly showed no significant modification or even a downregulation of the above-mentioned biomarkers. This latter finding demonstrated no additional effect after the cotreatment with both drugs with respect to the single treatments alone. Further studies are necessary to clarify the specific mechanism evoked by the processing of these neuroprotective agents in our in vitro models. Finally, these preliminary findings may represent a good tool with which to clarify the antioxidant and metabolic roles played by lipoic acid in proliferating and differentiating astroglial cell cultures, during an interactive cross-talk between glial and neuronal cells, after brain lesions or damage correlated with oxidative stress that may occur in some degenerative diseases.

Is there a role for video-assisted parathyroidectomy in regions with high prevalence of goitre?

Minimally-invasive procedures for parathyroidectomy have revolutionized the surgical treatment of primary hyperparathyroidism (pHPT). Coexistence of goitre is considered a major contraindication for these approaches, especially if unilateral. A specific advantage of video-assisted parathyroidectomy (VAP) compared to other endoscopic techniques is the possibility to combine it with thyroidectomy when necessary and when the selection criteria for video-assisted thyroidectomy (VAT) are met. We evaluated the role of VAP in a region with a high prevalence of goitre. The medical records of all patients who underwent parathyroidectomy and concomitant thyroid resection in our Division, between May 1998 and June 2012, were reviewed. Patients who underwent VAP and concomitant VAT were included in this study. Overall, in this period, 615 patients were treated in our Division for pHPT and 227 patients (36.9%) underwent concomitant thyroid resection. Among these, 384 patients were selected for VAP and 124 (32.3%) underwent concomitant VAT (lobectomy in 26 cases, total thyroidectomy in 98). No conversion to conventional surgery was registered. Mean operative time was 66.6 ± 43.6 min. Transient hypocalcaemia was observed in 42 cases. A transient recurrent nerve lesion was registered in one case. No other complications occurred. Final histology showed parathyroid adenoma in all but two cases of parathyroid carcinoma, benign goitre in 119 cases and papillary thyroid carcinoma in the remaining 5 patients. After a mean follow-up of 33.2 months, no persistent or recurrent disease was observed. In our experience, a video-assisted approach for the treatment of synchronous thyroid and parathyroid diseases is feasible, effective and safe at least considering short-term follow-up.

Are brown tumours a forgotten disease in developed countries?

Brown tumours (BT), an expression of osteitis fibrosa cystic due to primary hyperparathyroidism (pHPT), can occasionally be mistaken for malignancy. Among 615 patients who underwent parathyroidectomy for pHPT in our institution, the medical records of three patients affected by BT were reviewed. The first patient underwent surgical removal of the orbital mass for a suspected lachrymal gland neoplasm. The remaining two patients underwent, respectively, leg amputation and femur resection for a suspected bone malignancy. Final histology showed a BT in three cases. All three patients were admitted to our Division and underwent successful parathyroidectomy for parathyroid adenoma in two cases and for parathyroid carcinoma in the remaining case. When faced with an osteolithic bone lesion, complete evaluation of medical history, biochemical and radiographic findings can help to reach a correct diagnosis and avoid unnecessary bone resections.

Minimally-invasive parathyroid surgery.

During the last two decades, several techniques for minimally-invasive parathyroidectomy have been developed, including open approaches (open minimally-invasive parathyroidectomy - OMI P), minimally-invasive radio-guided parathyroidectomy (MI-RP), video-assisted parathyroidectomy (VAP), video-assisted parathyroidectomy through a lateral approach (VAP-LA) and purely endoscopic parathyroidectomy (EP). We have reviewed the pertinent literature, analyzing the indications, outcomes, advantages and disadvantages of the different techniques. Even if the field of minimally-invasive parathyroidectomy is heterogeneous, there is some evidence that minimally-invasive video-assisted parathyroidectomy (MIVAP) should be preferred over OMIP for better cosmetic outcomes, improved visualization of neck structures and control of pain. There is also low-level evidence that MIVAP has some advantages over other purely endoscopic procedures for parathyroidectomy and VAP-LA, in terms of technical difficulties, in addition to the possibility to perform bilateral exploration and associated procedures on the thyroid gland. While the data on medium-term results are encouraging, longer follow-up times are still needed to confirm its safety and rate of cure with respect to conventional surgery. It has been demonstrated that MIVAP is also feasible in secondary and familial hyperparathyroidism, although no conclusive data are available.

[Complications in thyroid surgery]. / Complicanze in chirurgia tiroidea.

Thyroidectomy is one of the most frequently performed surgical procedure worldwide, even if the risks of lethal postoperative complications prevented its evolution and diffusion until the beginning of the XX century. At that time, T. Kocher described his meticulous technique, reporting excellent results in terms of mortality and morbidity. At present, mortality for this procedure approaches 0% and overall complication rate is less than 3%. Nonetheless, major complications of thyroidectomy (i.e. compressive hematoma, recurrent laryngeal nerve palsy and hypoparathyroidism) are still fearful complications and account for a significant percentage of medico-legal claims. Patients volume and surgical skill play an important role in reducing the risk of complications. Accurate knowledge of anatomy and pathophysiology, complications incidence and pathogenesis and a careful surgical performance are essential. In this review, post-thyroidectomy complications basing on literature analysis and personal experience are described. The main anatomical, technical and pathophysiological factors that help preventing post-thyroidectomy complications are analyzed, taking into proper account new technologies and the minimally invasive surgical procedures that influenced thyroid surgery during the last decade.

Gene expression profiling of adrenal cortical tumors by cDNA macroarray analysis. Results of a preliminary study.

Adrenocortical carcinoma (ACC) are highly malignant tumors with poor prognosis. To verify if it is possible to assess their differential gene expression by a cDNA macroarray analysis using RNA extracted from paraffin sections, we analyzed two different cohorts of adrenal cortical adenoma (ACA) and ACC. Paraffin sections of seven ACC and seven ACA were analyzed. Transcriptional profiles were generated by commercially available c-DNA arrays testing 82 genes. Hybridization signals were quantified by densitometry and the intensity signal was compared for each gene between ACA and ACC cohorts. RNA was successfully extracted in only four out of 14 cases. Four genes displayed a significantly different expression (ACC/ACA ratio>1.5 or<0.6). Heat shock protein 60 (HSP-60) (ratio>2), Ciclin D1 and topoisomerase I (ratio>1.5) were overexpressed in the ACC cohort, while jun proto-oncogene was down-regulated. cDNA macroarray analysis from paraffin sections of adrenal tumors is feasible, despite with a low success rate. The different expression of HSP-60, Ciclin D1, jun proto-oncogene and topoisomerase I indicates that these genes may play a role in ACC pathogenesis and could represent potential diagnostic/prognostic/therapeutic target markers. Larger series of patients are necessary to confirm the biologic, diagnostic, prognostic and therapeutic implications of these findings.

Dopamine plasma membrane transporter (DAT) in rat thymus and spleen: an immunochemical and immunohistochemical study.

The expression of the dopamine plasma membrane transporter (DAT) was investigated in rat thymus and spleen by immunochemical and immunohistochemical techniques. Antibodies raised against a peptide mapping near the amino terminus of DAT were bound to a single band of approximately 76 kDa in thymus and spleen membranes as well as in striatal and kidney membranes which were used as dopaminergic reference tissues. Reverse transcription-polymerase chain reaction analysis revealed that both thymus and spleen expressed DAT mRNA. Immunohistochemistry revealed in rat thymus a DAT immune reaction in the wall of arteries located in septa of connective tissue as well as in the medulla, with a reticular localization and an apparent negative reaction of thymocytes. In the spleen, DAT immunoreactivity was located primarily in the red-white pulp marginal zone, within small cells, likely corresponding to lymphocytes and in the wall of white pulp arteries. The presence of a dopamine transporter suggests that dopamine released in the lymphoid microenvironment may contribute to neuroimmune modulation. It cannot be excluded a different activity of dopamine in primary and secondary immune organs, such as maturation and selection of lymphocytes and activation of immune responses in the spleen.

[Pheochromocytoma: role of preoperative diagnosis in the assessment of malignancy risk and in the choice of surgical approach]. / Feocromocitoma: ruolo della diagnostica preoperatoria nella valutazione del rischio di malignita e nella scelta dell'approccio chirurgico.

Pheochromocytomas are malignant in 5-26% of the cases. Differential diagnosis with benign lesions can be difficult even on a pathological basis. Local invasion and distant metastasis are the only well established indicators of malignancy. It has been reported that the risk of malignancy increases with the lesion size. Despite safe laparoscopic adrenalectomy (LA) has been reported for lesions up to 10 cm, it is considered hazardous for pheochromocytoma larger than 6 cm, because of the risk of malignancy and iatrogenic pheochromocytomatosis. We evaluated the possibility to pre-operatively recognize pheochromocytomas at risk of being malignant that should not be selected for LA. The medical records of all the patients who underwent adrenalectomy for pheochromocytoma were reviewed. All the preoperatively available data (demographic, clinical, biochemical and radiological) were recorded as well as final pathological diagnosis. Comparative analysis of patients with benign and malignant pheochromocytomas was performed. Sixty-three adrenalectomies for pheochromocytoma were performed in 60 patients. Fifty-seven benign and 6 malignant pheochromocytomas were identified. No significant difference was found between patients with malignant and benign lesions concerning age, gender, family history, symptoms, laboratory and radiological findings. In particular, no significant difference was found for lesion size between the benign (63.3 +/- 30.6 mm, range, 20-150) and the malignant group (48.6 +/- 16.5 mm; range, 30-70). The largest diameter recorded for a malignant lesion was 70 mm. No preoperatively available data can reliably differentiate between benign and malignant pheochromocytomas. All malignant lesions in this series were smaller than 7 cm. Thus, pheochromocytoma size does not seem a reliable predictor of malignancy. In absence of the evidence of gross local invasion or metastatic disease, LA can be safely proposed also for large lesions. Conversion is mandatory in presence of local invasion or difficult dissection that could involve inadequate resection.

[Laparoscopic adrenalectomy in the treatment of malignant adrenal lesions]. / La surrenectomia laparoscopica nel trattamento delle lesioni surrenaliche maligne.

The role of laparoscopic approach for the treatment of malignant adrenal diseases is still controversial. The aim of this study was to verify the results of laparoscopic adrenalectomy in the management of adrenal malignancies. The medical records of all the patients who underwent laparoscopic procedures for adrenal diseases and in whom malignancy was demonstrated at final histology were reviewed. Nine patients were included (3 malignant pheochromocytomas, 4 adrenocortical carcinomas and 2 adrenal metastases). At a mean follow-up of 17.0 +/- 12.8 months (range, 2-36), all but two patients were alive and disease free. One patient died for unrelated causes. No patient developed local or port site recurrence. The results of this study demonstrate that laparoscopic adrenalectomy can be safe and effective also in case of adrenal malignancies. Conversion to open surgery in mandatory in case of local invasion and when the dissection cannot be as accurate as in conventional operations. A preliminary laparoscopic exploration can be planned in case of suspected malignant lesions to confirm the diagnosis and to evaluate their operability.

Video-assisted thyroidectomy for papillary thyroid carcinoma.

BACKGROUND: In patients with small papillary thyroid carcinomas (PTC), we evaluated the operative feasibility and safety of video-assisted thyroidectomy (VAT) and the completeness of the surgical resection. METHODS: Video-assisted thyroidectomy was attempted in 24 patients with thyroid malignancy. Total thyroid resection for PTC was achieved completely by VAT in 20 of them, who were included in this study. RESULTS: In this study, 12 total thyroidectomies and 8 lobectomies followed by completion thyroidectomies were performed. Eight patients also underwent central neck lymph node dissection. Mean postoperative serum thyroglobulin was 0.2 ng/ml for patients receiving LT4 suppressive treatment and 4.2 ng/ml for patients after LT4 withdrawal. Postoperative ultrasonography showed no residual thyroid tissue. The mean radioiodine uptake at postoperative scintiscan was 2.2%. CONCLUSIONS: In the case of PTC, VAT is feasible and safe. The completeness of the surgical resection seems comparable with that reported for conventional surgery. Nevertheless, larger series and longer follow-up evaluation are necessary for definitive conclusions to be drawn about its oncologic validity.

Sulfatides and arylsulfatase A activity in major salivary glands of hamster (Mesocricetus auratus) after adenocarcinoma induction in oral cavity.

A biochemical study of sulfatides and arylsulfatase A (ASA) was carried out in the submandibular and sublingual glands of the male and female hamster Mesocricetus auratus after experimental induction of oral adenocarcinoma by 7,12-dimethylbenzanthracene (DMBA). Hamster experimental groups included control animals, animals treated with beta-carotene, animals treated with DMBA, and animals treated with DMBA plus beta-carotene. Oral cavity treatment with DMBA induced carcinogenesis in the buccal mucosa, but not in the major salivary glands, where nevertheless, the morphology and expression of both parameters examined changed. In fact, sulfatide concentrations and enzyme activity increased significantly, while in control and beta-carotene-treated hamsters they were similar in both glands and sexes. After administration of DMBA plus beta-carotene, sulfatide concentration decreased, as did ASA activity, slightly in the submandibular gland and remarkably so in the sublingual one of female hamsters. Thin-layer chromatography (TLC) analysis of lipid patterns, after DMBA treatment, revealed considerable differences, not only in sulfatides, but also in other lipid fractions, as well as between the two glands and two sexes. These findings show that oral cavity treatment with DMBA is not able to induce carcinogenesis in the major salivary glands examined; however, it does cause considerable metabolic changes.

Is the identification of the external branch of the superior laryngeal nerve mandatory in thyroid operation? Results of a prospective randomized study.

BACKGROUND: The aim of this study was to investigate the incidence of injury to the external branch of the superior laryngeal nerve (EBSLN) with 2 different surgical approaches. METHODS: From 1998 to 2000, 289 consecutive patients undergoing thyroidectomy were randomly divided into 2 groups. In group A (137 patients [215 upper pole ligations]), the superior thyroid artery was ligated after identification of the EBSLN. In group B (152 patients [244 upper pole ligations]), the superior thyroid artery's branches were ligated separately close to the gland. In all patients, a phoniatric evaluation with videostrobolaryngoscopy and spectrographic examination was performed. RESULTS: The 2 groups were well matched regarding age, sex, thyroid pathological findings, and type of operation. In group A, the EBSLN was not clearly identified in 11.6% of cases. Alterations of EBSLN function were absent in both groups of patients, either postoperatively or 1 and 6 months after operation. Group B showed statistically significant shorter operative time compared with that for group A. CONCLUSIONS: Even if the EBSLN often crosses the superior thyroid pedicle, especially in large goiters, this study demonstrated that accurate distal ligation of the branches of the superior thyroid artery is a safe technique to prevent EBSLN injury.

[Assessment of the adequacy of health services in day hospital care: experience of an endocrine surgery day care unit at a university polyclinic]. / Valutazione di appropriatezza delle prestazioni sanitarie effettuate in regime di Day Hospital: l'esperienza di una unità di DH di chirurgia endocrina in un Policlinico Universitario.

The aim of the study was to assess the appropriateness of the provision of surgical treatment in the day hospital setting and at the same time to evaluate the efficacy of the service provided. The study was conducted on admissions to a day care unit operating in conjunction with the surgery division of a university hospital. The evaluation of appropriateness was carried out using explicit criteria, based on regional regulations and on guidelines drawn up by the Agency for Regional Health Services. The criteria related to the hospital bed equivalent Rotation Index and to the types of DRGs treated and services provided. Efficacy was assessed using the following para-meters: number of US-guided fine needle aspiration biopsies; techniques used; age, sex and nodule size; patient distribution by results of diagnostic examination; cytological classification; percentage of patients with complications. We also evaluated the possibility of transferring short-term (2-3 days) surgical admissions from ordinary regimens to the day care setting. The results of this study yield useful synthetic indicators for assessing the appropriateness both of the day care function as a whole and of individual operating units, providing both administrative and medical staff with a useful frame of reference for the planning of health-care management.

Central nervous system involvement during infection by Phlebovirus toscana of residents in natural foci in central Italy (1977-1988).

Central nervous system (CNS) involvement was detected during infection caused by the sand fly-transmitted Phlebovirus Toscana. One hundred fifty-five cases of Toscana virus-associated meningitis or meningoencephalitis were identified in a survey that lasted ten years, conducted in two regions of central Italy. Diagnosis was performed by different serologic tests. A combination of hemagglutination-inhibition and plaque-reduction neutralization or indirect immunofluorescence for IgM, and enzyme-linked immunosorbent assays for IgM were considered the most suitable tests for the diagnosis of Toscana virus infection. A few strains of Toscana virus were isolated from the cerebrospinal fluid of seropositive patients. Toscana virus-associated CNS disease occurred during the summer, reaching a peak value in August, when the maximum activity of the sand fly vector occurs and virus isolates are obtained in their natural foci. The results suggest that Toscana virus should be considered as a possible cause of CNS disease in Mediterranean countries where sand flies of the genus Phlebotomus are known to be present.