Role of thyroid stimulating hormone in the maintenance and functioning of the human corpus luteum.

PURPOSE: To evaluate the impact of high thyroid stimulating hormone (TSH) levels on human granulosa-luteal (hGL) cells. METHODS: hGL cells were isolated from follicular aspirates derived from patients undergoing IVF treatment without any thyroid disorder (serum TSH 0.5-2 mU/L). Cells were cultured at 37 °C in DMEM, supplemented with 5% FBS. The cells were treated with 1 nM LH and increasing concentrations of TSH. At the end of culture, conditioned medium and cells were collected to analyze progesterone production, cell viability, and mRNA levels of genes involved in the steroidogenesis process. Human ovarian tissues were analyzed for TSH receptor (TSHR) expression by IHC. RESULTS: The expression of TSHR was detected in human corpus luteum by IHC and in hGL by RT-PCR. In hGL cells, TSH treatment did not modulate progesterone production nor the expression of steroidogenic genes, such as p450scc and HSD3b 1/2. However, TSH induced a dose-dependent increase in cell death. Finally, TSH did not affect LH-induced p450scc and HSD3b1/2 expression while LH partially reverted TSH negative effect on cell death in hGL. CONCLUSIONS: Elevated TSH levels in hypothyroid women may be associated with impaired CL functioning and maintenance. These findings open a new line of research for the importance of the treatment of women with thyroid dysfunction that could contribute to the onset of infertility.

Nanocomposite hyaluronic acid-based hydrogel for the treatment of esophageal fistulas.

Fistulas are abnormal connections between two body parts that can impair the quality of life. The use of biological glues represents the least invasive procedure to fill the fistula; however, it is limited by the need of multiple injections, the persistence of infection and the failure in the treatment of high-output fistulas. We describe herein the use of an injectable nanocomposite hydrogel that is able to form in situ a tissue-mimicking matrix as an innovative material for the treatment of esophageal fistulas. Injectable hydrogels that have the dual advantage of being implantable with a minimally invasive approach and of adapting their shape to the target cavity, while the introduction of mesoporous silica nanoparticles opens the possibility of drug/biomolecules delivery. The hydrogel is based on hyaluronic acid (HA), the crosslinking process occurs at physiological conditions leading to a hydrogel made of >96% by water and with a large-pore micro-architecture. The kinetic profile of the hydrogel formation is studied as a function of HA molecular weight and concentration with the aim of designing a material that is easily injectable with an endoscopic needle, is formed in a time compatible with the surgical procedure and has final mechanical properties suitable for cell proliferation. The in vivo experiments (porcine model) on esophageal-cutaneous fistulas, showed improved healing in the animals treated with the hydrogel compared with the control group.

"Hormone of darkness" and human reproductive process: direct regulatory role of melatonin in human corpus luteum.

PURPOSE: To investigate the possible role of melatonin on human luteal cell function. METHODS: Corpora lutea were obtained from normally menstruating women (25-38 years old) in the midluteal phase (days 5-6 from ovulation) at the time of surgery for non-endocrine gynecologic diseases. The protocol was approved by the institutional review board of Università Cattolica del Sacro Cuore in Rome and all patients provided written informed consent. The corpora lutea were dated on the basis of the presumptive day of ovulation (day 0) , determined by urinary luteinizing hormone (LH) peak, ultrasound detection of corpus luteum or disappearance of the dominant follicle, and a rise in the plasma P concentration. ELISA or EIA kit and immunohistochemistry were performed. RESULTS: Melatonin was able to increase progesterone release and to influence the balance between luteotrophic and luteolityc factors. In addition, melatonin expression and MT2 receptor were detected, confirming the direct action of this indoleamine on CL. CONCLUSIONS: Melatonin may play an intriguing role in direct regulation of CL function and in establishing and maintaining of initial pregnancy. In conclusion, melatonin could become a relevant medication for improving ovarian and luteal function and in the early stages of pregnancy, opening new opportunities for the management of several ovarian-luteal and pregnancy diseases.

The role of high-mobility group box protein 1 in collagen antibody-induced arthritis is dependent on vascular endothelial growth factor.

High-mobility group box 1 (HMGB1) has been implicated in angiogenesis and rheumatoid arthritis (RA). The aim of this study was to define more clearly the role of HMGB1 in the synovial angiogenesis and pathogenesis of an immune model of arthritis. BALB/c mice were injected with monoclonal anti-collagen antibody cocktail followed by lipopolysaccharide to induce arthritis. HMGB1 and vascular endothelial growth factor (VEGF) were over-expressed in the areas of the synovium where more inflammation and neoangiogenesis were present. The selective blockade of HMGB1 or VEGF resulted alternatively in a lower severity of arthritis evaluated by the arthritis index. Furthermore, exogenous HMGB1 administration caused a worsening of arthritis, associated with VEGF up-regulation and increased synovial angiogenesis. The selective inhibition of VEGF also resulted in no induction of arthritis in mice receiving exogenous HMGB1. Cytokine enzyme-linked immunosorbent assay (ELISA) analyses performed on peripheral blood and synovial fluid demonstrated a significant reduction of interleukin (IL)-1ß, IL-6 and tumour necrosis factor (TNF)-α in mice where HMGB1 and VEGF pathways were blocked. Interestingly, the selective blockade of HMGB1 and VEGF resulted in an increase of the peripheral IL-17A concentration. The development of arthritis mediated by HMGB1 and the synovial angiogenesis can be blocked by inhibiting the VEGF activity. The proinflammatory and proangiogenic cytokine IL-17A was increased when HMGB1 is inhibited, but the synovial angiogenesis was nevertheless reduced in this model of arthritis. Taken together, these findings shed new light on the role of this nuclear protein in the pathogenesis of arthritis in an RA-like model.

18F-FDG PET/CT in the post-operative monitoring of patients with adrenocortical carcinoma.

CONTEXT: The role of (18)F-labeled 2-fluoro-2-deoxy-d-glucose (FDG) positron emission tomography (PET)/computed tomography (CT) in the post-operative monitoring of patients with adrenocortical carcinoma (ACC) is still unclear. OBJECTIVE: To assess the accuracy of FDG PET/CT to diagnose ACC recurrence in a real world setting. DESIGN AND METHODS: Retrospective evaluation of data of 57 patients with presumed ACC recurrence at CT scan who underwent FDG PET/CT within a median time of 20 days. We compared the results of either FDG PET/CT or CT with a gold standard confirmation of recurrence (positive histopathology report of removed/biopsied lesions or radiological progression of target lesions at follow-up) to assess their diagnostic performance at different body sites to correctly categorize target lesions. We also assessed whether FDG PET/CT findings may be useful to inform the management strategy. RESULTS: In 48 patients with confirmed ACC recurrence, we found that FDG PET/CT had lower sensitivity than CT in diagnosing liver and lung recurrences of ACC. FDG PET/CT had higher specificity than CT in categorizing liver lesions. FDG PET/CT had a greater positive likelihood ratio than CT to identify liver and abdominal ACC recurrences. The management strategy was changed based on FDG PET/CT findings in 12 patients (21.1%). CONCLUSIONS: The greater sensitivity of CT may be partly expected due the specific inclusion criteria of the study; however, the greater specificity of FDG PET/CT was particularly useful in ruling out suspected ACC recurrences found by CT. Thus, use of FDG PET/CT as a second-line test in the post-operative surveillance of ACC patients following CT finding of a potential recurrence may have a significant impact on patient management.

The heme oxygenase/biliverdin reductase system in skin cancers.

The heme oxygenase/biliverdin reductase (HO/BVR) pathway enhances cell stress response by degrading excess heme or producing antioxidant and cytoprotective molecules. Recently, members of the HO/BVR system have been proposed as biomarkers for the early diagnosis of free radical-related diseases. In this study, the presence of both the inducible and constitutive HO isoforms (HO-1 and HO-2, respectively) and BVR was evaluated by immunohistochemistry in human skin cancer samples. Moderate/strong immunoreactivities against HO-1, HO-2 and BVR were detected in 100% of the nodular malignant melanoma samples, whereas in basal cell carcinoma specimens these figures were 62%, 88% and 60%, respectively, with a faint/moderate degree of expression. Faint/moderate HO-1, HO-2 and BVR immunoreactivities were detected in 33%, 66% and 100% of melanocytic nevi samples, respectively. In conclusion, HO-1 and HO-2 and BVR were expressed in the cytosols of skin cancer cells, whereas perilesional normal epidermis showed only faint staining, thus leading to the hypothesis that the HO/BVR system is activated in skin cancers.

Haemophagocytic syndrome associated with mucormycosis infection.

Several clinical forms of mucormycosis are recognized. The tendency of mucoraceous zygomycetes to invade the blood vessels often produces a disseminated infection. A case of disseminate mucormycosis complicated by a haemophagocytic syndrome (HS) in a 32-year-old Caucasian male is reported in this article. Few cases of infection-associated HS (IAHS), involving infections caused by fungi, have been reported. In all the recorded cases, the fungal infection coexists with malignant lymphoma, immunodeficiency and a long-term steroid therapy for renal transplant or Crohn's disease. This is the second described case of the HS due to mucormycosis.

Role of integrated PET/CT with [¹8F]-FDG in the management of patients with fever of unknown origin: a single-centre experience.

PURPOSE: Fever of unknown origin (FUO) is a diagnostic challenge for the wide range of possible causes involved. The aim of our work was to evaluate the role of [(18)F]-fluorodeoxyglucose positron emission tomography computed tomography ([¹8F]-FDG-PET/CT) in managing patients with classical FUO. MATERIALS AND METHODS: Twenty-four consecutive patients (16 women, eight men; mean age, 56.5 years) with a diagnosis of FUO based on routine investigations were retrospectively studied. All underwent [¹8F]-FDG-PET/CT, which was considered true positive when the result was in agreement with the final diagnosis. RESULTS: A final diagnosis was reached in 17 of 24 patients (vasculitis, n=5; autoimmune disorder, n=2; neoplasm, n=3; infectious disease, n=6; biliary microlithiasis, n=1). In the remaining seven cases, no final diagnosis was established. PET-CT was useful in identifying aetiology in 11 patients, showing a diagnostic yield of 46% (11/24). Among the 11 cases with a negative PET scan, 10 did not show a worsening of the clinical condition. CONCLUSIONS: This study underlines the crucial role of [¹8F]-FDG-PET/CT in managing patients with FUO. If prospective trials on this topic confirm the present findings, PET/CT should be incorporated in the routine diagnostic work-up of patients with classical FUO.

Prognostic role of the PET parameter maximum standardized uptake value in non small cell lung cancer: analysis in tumour of diameter ≥ and <25 mm.

AIM: The aim of this study was to evaluate whether the primary tumour maximum standardized uptake value (SUV(max)) plays an independent prognostic role in patients with non small cell lung cancer (NSCLC) and whether this role is limited by partial volume effect (PVE) and motion artefacts. METHODS: One hundred and fifty-three consecutive patients underwent PET exam, surgery (R0 resection) and follow-up (mean 20.3; range 6-44.8 months). Correlation with Disease Free and Overall Survival (DFS, OS) was evaluated in the entire population for: SUV(max), clinical and histopathological features and pathological stage. To evaluate the PVE and motion artefacts' interferences on SUV calculation, the correlation between SUV(max) and DFS/OS was also calculated in the groups of patients with tumour diameter ≥ and < than 25 mm (group A and B, respectively). RESULTS: In the entire population only TNM and SUV(max) resulted correlated with DFS/OS. However, SUV(max) was significantly correlated with DFS/OS in group A but not in group B. Furthermore, only in the group of patients with primary tumour diameter ≥ 25 mm (group A), tumour diameter, tumour histotype, and tumour necrosis resulted significantly related with SUV(max) at both uni and multivariate analysis. CONCLUSION: TNM together with SUV(max) could be useful in giving a better prognostic stratification of patients with NSCLC; however technical limitations in the SUV calculation must be taken into account in patients with tumour diameter <25 mm.

Bone marrow disease detection with FDG-PET/CT and bone marrow biopsy during the staging of malignant lymphoma: results from a large multicentre study.

AIM: To compare the accuracy of bone marrow biopsy (BMb) and positron emission tomography (PET) in bone marrow disease (BMD) detection, in a large multicentre population of patients with new diagnosis of malignant lymphoma. METHODS: PET and BMb were performed to complete disease staging in 337 consecutive patients: 130 Hodgkin's disease (HD), 207 aggressive non Hodgkin's lymphoma (NHL). Sensitivity, specificity and accuracy of both techniques in BMD detection were evaluated and compared. RESULTS: 87 patients with BMD (25 positives at both exams, 27 only at the BMb and 35 only at the PET study). PET vs. BMb were reordered: sensitivity: 69% vs. 59.8%; specificity: 99.2% vs. 100%; accuracy: 91.4% vs. 89.6%; positive predictive value: 96.8% vs. 100%; negative predictive value: 90.2% vs. 87.7%. CONCLUSION: The sensitivity of PET and BMb is similar (69% and 60%, respectively), PET and BMb are complementary: in fact out of 87 patients with confirmed BMD only 25 are positive at both exams, while 27 only at the BMb and 35 only at the PET exam; the integration of PET findings with BMb ones increases the diagnostic accuracy. Consequentially PET is essential during the staging of malignant lymphomas.

Aggressive large B-cell lymphoma in a systemic lupus erythematosus patient with chronic active Epstein-Barr virus infection: a case report.

A link between Epstein-Barr Virus (EBV) infection, systemic lupus erythematosus (SLE) and non-Hodgkin's lymphoma (NHL) has been recently reported in literature. Here we report a case of diffuse large B-cell lymphoma (DLBCL) with a particularly aggressive clinical course in an SLE patient with EBV infection. A 49-year-old woman with a long history of SLE was admitted to the Department of Experimental and Clinical Medicine and dramatically died a few hours later. The autopsy described no evidence of active lymphoproliferative disorder. Instead, histological examination demonstrated an atypical lymphocitic proliferation in lymph node, kidneys, pericardium and uterus. Immunoistochemically, the lymphomatous cells were positive with CD19, CD20, CD22 and CD79a, which was consistent with a DLBCL. The cells were also reactive to EBV markers, indicating the possible role of previous EBV infection in DLBCL pathogenesis.

Dual-phase FDG-PET: delayed acquisition improves hepatic detectability of pathological uptake.

PURPOSE: The aim of this study was to evaluate whether the acquisition of delayed images could improve the detectability of liver pathological uptakes. MATERIALS AND METHODS: Ninety-five consecutive patients with suspected liver metastases underwent a dual-phase positron emission tomography (PET) scan. All patients underwent a whole-body PET/computed tomography (CT) scan (PET-1) acquired 1 h post [18F]fluorodeoxyglucose (FDG) injection, and a liver PET/CT scan [that is, one or two fields of view (FOV) of the upper abdomen; PET-2] acquired 2 h postinjection. In all cases, image reconstruction was performed as 3D reconstruction algorithm Fourier rebinning (FORE) iterative, FOV 50 cm, image matrix size 128 x 128. Both studies were evaluated qualitatively and semiquantitatively [background standard uptake values (SUV)mean of the liver, lesion SUVmax and SUVmean and ratio SUVmean lesion/background). RESULTS: Thirty-seven of 95 patients (38.9%) presented liver lesions at both PET-1 and PET-2 exams, whereas there were two (2.2%) only at PET-2. Eighty-one liver lesions were identified at both PET studies, whereas there were nine (11.1%) only at PET-2. Furthermore, at PET-2, we had a statistically significant reduction of SUVmean background values (p < 0.001, Wilcoxon test) and a concomitant increase of SUVmean lesion values (p < 0.001, Wilcoxon test), ratio lesion to background (p < 0.001, Wilcoxon test). CONCLUSIONS: Acquisition of delayed images improved the hepatic detection of pathological FDG uptake.

Role of whole-body 18F-choline PET/CT in disease detection in patients with biochemical relapse after radical treatment for prostate cancer.

PURPOSE: The aim of this study was to evaluate the role of whole body 18F-choline (FCH) positron emission tomography-computed tomography (PET-CT) in detecting and localising disease recurrence in patients presenting biochemical relapse after radical treatment for prostate cancer. MATERIALS AND METHODS: Fifty-six consecutive patients with increased serum prostate-specific antigen (PSA) levels after radical prostatectomy were included in the study. None of them was receiving hormone treatment at the time of the examination or had been treated during the previous 6 months. All patients underwent whole-body 18F-choline PET imaging, and the pathological findings were compared with those of further imaging exams, biopsy and follow-up. On the basis of the PSA levels, we divided our patient population into three subgroups: PSA < or = 1, 1 < PSA < or = 5, and PSA > 5 ng/ml. RESULTS: Overall, the PET scan detected disease relapse in 42.9% of cases (24/56). PET sensitivity was closely related to serum PSA levels, showing values of 20%, 44% and 81.8% in the PSA < or = 1, 1 < PSA < or = 5 and PSA > 5 ng/ml subgroups, respectively. CONCLUSIONS: In patients with biochemical relapse after radical treatment for prostate cancer, 18F-choline PET-CT represents a single step, whole-body, noninvasive study that allows disease detection and localisation. The disease detection rate is related to serum PSA levels.

Borderline HER-2 breast cancer cases: histochemical versus real-time PCR analysis and impact of different cut-off values.

Seventy-one cases that had resulted borderline for HER-2 protein expression at conventional immunohistochemical assay (2+) were assessed for HER-2 gene amplification by real-time PCR and by FISH in accordance with the manufacturer's recommendations (gene amplification with ratio >or=2 in both methods). Thirty-three out of 71 cases (47%) resulted amplified at real-time PCR analysis, whereas 15 cases resulted positive at FISH (21%). Apparently, PCR was more sensitive than FISH in HER-2 determination, only 10 cases resulting amplified in both tests. When the mean ratio value obtained in all PCR experiments was adopted as threshold in determining HER-2 gene amplification, the apparent sensitivity of PCR was reduced but correlation between PCR and FISH results was dramatically increased. Furthermore, when the mean PCR ratio value observed in the FISH-positive group was chosen as threshold, the best agreement between PCR and FISH results was achieved. Therefore, we found that the proposed threshold ratio value of >or=2 is not accurate in separating HER-2 amplified and non-amplified cases. We suggest that the threshold ratio value in PCR tests should be determined in each laboratory using FISH controlled cases. Finally, above certain in-lab generated threshold values, PCR might be proposed as a highly predictive positive test in HER-2 assessment.