Association among 1267 A/G HSP70-2, -308 G/A TNF-α polymorphisms and pro-inflammatory plasma mediators in old ZincAge population.

Proinflammatory cytokines and heat shock proteins play relevant roles in the pathogenesis of inflammatory diseases. We investigated whether Hsp70 1267 A/G and TNF-α -308 G/A polymorphisms are associated with proinflammatory mediators, zinc status and laboratory parameters in 1,078 healthy elderly from ZincAge study. Hsp70 1267 A/G genotype and allele distribution were similar among various European countries, while a TNF-α genetic heterogeneity was observed between the Northern and the Southern European populations, with a major frequency of the -308 A variant in France, Germany and Poland. We used linear regression models to test additive, dominant or recessive associations of each SNP with proinflammatory mediators, laboratory parameters, metallothioneins and zinc status. Hsp70 1267 A/G SNP, but not TNF-α -308 G/A SNP, influences TNF-α and IL-6 plasma levels under additive, dominant and recessive models (for TNF-α only). An association between Hsp70 1267 A/G SNP and zinc plasma levels was observed in the dominant model. In particular, G allele carriers showed increased circulating pro-inflammatory cytokines and zinc. Moreover, both these SNPs affect creatinine levels suggesting a possible influence on renal function. In conclusion, Hsp70 1267 A/G SNP is associated with pro-inflammatory cytokine production in healthy elderly and might represent a possible determinant of individual susceptibility to inflammatory diseases.

Zinc, metallothioneins and immunosenescence.

Ageing is an inevitable biological process with gradual and spontaneous biochemical and physiological changes and increased susceptibility to diseases. The nutritional factor, zinc, may remodel these changes with subsequent healthy ageing, because zinc improves the inflammatory/immune response as shown by in vitro and in vivo studies. The intracellular zinc homeostasis is regulated by buffering metallothioneins (MT) and zinc transporters (ZnT and ZIP families) that mediate the intracellular zinc signalling assigning to zinc a role of 'second messenger'. In ageing, the intracellular zinc homeostasis is altered, because high MT are unable to release zinc and some zinc transporters deputed to zinc influx (ZIP family) are defective leading to low intracellular zinc content for the immune efficiency. Physiological zinc supplementation in the elderly improves these functions. However, the choice of old subjects for zinc supplementation has to be performed in relation to the specific genetic background of MT and IL-6, because the latter is involved both in MTmRNA and in intracellular zinc homeostasis. Old subjects carrying GG genotypes (C-carriers) in the IL-6-174G/C locus display high IL-6, low intracellular zinc content, impaired innate immunity and enhanced MT. Old subjects carrying GC and CC genotypes (C+carriers) display satisfactory intracellular zinc content, adequate innate immunity and are more prone to reach longevity. Zinc supplementation in old C-carriers restores natural killer cell cytotoxicity and zinc status. The genetic variations of the IL-6-174G/C locus when associated with those of the MT1A+647A/C locus are useful tools for the choice of old people for zinc supplementation.

Effects of interleukin-6 -174C/G and metallothionein 1A +647A/C single-nucleotide polymorphisms on zinc-regulated gene expression in ageing.

Decreased zinc ion availability in ageing is associated with altered immune response. One of the main regulators of zinc availability is metallothionein. Metallothionein induction is under the control of interleukin-6, a pro-inflammatory cytokine whose production is associated with poor ageing. The production of interleukin-6 is controlled, in part, by variability in the -174 nucleotide position. Under conditions of chronic inflammation, such as in ageing, zinc release by metallothionein is limited and may reduce zinc availability. Understanding the precise nature of the interactions between interleukin-6 and metallothioneins will aid in identifying individuals who are at risk of zinc deficiency. In the current study, we used gene arrays to investigate the effects of in vitro zinc supplementation on gene expression in elderly donors with described interleukin-6 and metallothionein 1a polymorphisms. Ingenuity Pathway Analysis identified several zinc-responsive genetic networks uniquely regulated only in elderly individuals with the pro-inflammatory interleukin-6 polymorphism. These include zinc-dependent decreased transcription of pro-inflammatory cytokines and alterations in metabolic regulatory pathways. The genomic effects of zinc increased in significance in the presence of the metallothionein 1a +647 C/A transition, suggesting that the interleukin-6 and metallothionein 1a genes act in a concerted manner to control zinc-regulated gene expression.

+647 A/C and +1245 MT1A polymorphisms in the susceptibility of diabetes mellitus and cardiovascular complications.

Diabetes mellitus is a chronic disease characterized by an overproduction of reactive oxygen species, which perturbs zinc metabolism and promotes the onset of cardiovascular disease (CVD) in diabetic patients. Metallothioneins (MT) are cysteine-rich metal-binding proteins which, by means of their antioxidant and zinc-buffering properties, might prevent the development of diabetic cardiovascular complications. A recent investigation shows that a polymorphism (+647 A/C) in the human MT-1A gene, affects the intracellular zinc ion release (iZnR) from the proteins and is associated with longevity in Italian population. The aim of the present study is to assess the involvement of +647 A/C and +1245 A/G MT1A polymorphisms with the susceptibility to type 2 diabetes (DM2) and cardiovascular complications. The study included 694 old individuals: 242 old healthy controls, 217 DM2 patients without clinical evidence of CVD (DNC) and 235 diabetic patients with diagnosis of CVD (DCVD). +647 A/C MT1A polymorphism, but not the second SNP, was associated with DM2. C allele carriers were more prevalent in DNC and DCVD patients than in control group (OR=1.37, p=0.034; OR=1.54, p=0.002, respectively). C+ carriers was associated with higher glycemia and glycosylated hemoglobin in DCVD patients, but not in DNC or control subjects. No differences in plasma zinc, but a modulation of MT levels and iZnR in PBMCs were observed in DCVD cohort when related to +647 A/C MT1A polymorphism. In summary, this work provides novel evidence on the association of the +647 A/C MT1A polymorphism with DM2. Moreover, C+ carriers in DCVD patients presented a worse glycemic control, a reduced iZnR and a higher MT levels, suggesting a possible role of MT in diabetic cardiovascular complications.

A standardized sentinel lymph node enhanced pathology protocol (SEPP) in patients with breast cancer.

BACKGROUND AND OBJECTIVES: Sentinel lymph node (SLN) biopsy allows enhanced pathology with serial sections and immunohistochemical analysis of the retrieved node. We present our experience with a simple, practical, pathology protocol. METHODS: We analysed 416 consecutive breast cancer patient who underwent SLN biopsy. These were studied with six couples of sections at three different levels, each stained with hematoxylin-eosin (H/E) and immunohistochemistry (IHC) (MNF 116). RESULTS: With conventional analysis the SLN was positive in 106/416 cases (25%). The addition of serial sections, according to the protocol, allowed diagnoses of micrometastases (MICRO) (n = 22) or isolated tumor cells (ITC) (n = 38) or MICRO (n = 1) in 51/416 patients (14.6%). Specifically, the diagnosis was undertaken at level I (8.9%), level II (4%), or level III (1,6%). The incidence of MICRO or ITC was not different in T1 and T2 cases (13% vs. 15%, P = 0.7). The addition of the third level of analysis added very little both in T1 and T2 cancers (1.3% vs. 3.8%, P = 0.1). CONCLUSIONS: Serial sectioning of the SLN allows diagnosis of MICRO and ITC in a significant percentage of cases. Adoption of our protocol seems practical, as the incidence of level III positivity is extremely low, particularly in T1 cancers, and additional sections would be, therefore, unlikely useful.

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.

Haemothorax caused by isolated costal exostosis.

We report an unusual case of spontaneous haemothorax in a 13-year-old girl with isolated costal exostosis. Surgical excision of the exostosis was performed with complete resolution. Costal exostosis should be considered in the differential diagnosis of spontaneous haemothorax in children in order to avoid unnecessary investigation and to establish an adequate treatment plan.

Bone marrow and sentinel lymph node biopsy in patients with breast cancer: from staging to ultrastaging?

Bone marrow (BM) biopsy has been suggested as an independent prognostic tool to improve staging in patients with breast cancer. Two hundred and ten consecutive patients operated for breast cancer from June 2000 to June 2005 who signed an informed consent were enrolled in this protocol. Patients underwent SLN biopsy, and lymph nodes were analysed with serial sections and stained with hematossilin-eosin and immunohistochemistry. At the end of the procedure a BM aspirate from the iliac crest was obtained and 5-10 cc of blood collected. A CEA specific nested reverse transcriptase (RT) polymerase chain reaction (PCR) assay was used to examine BM samples. Results were blinded to both patients and clinicians. The median age of the patients was 56 years (range 34-80), and the median tumor diameter 1,5 cm (range 0.2-4.5). BM aspirates were unsuccessful in ten patients, and RT-PCR was not technically feasible in seventeen women, leaving 183 patients available for analysis of results and follow up. SLN biopsy allowed diagnoses of occult metastases (micrometastases and isolated tumor cells) in 16% of patients (29/183). 25% of T1N0 patients (23/92), 35% of T2N0 patients (6/17), and 44% of N1-2 patients (32/72) were BM+ (p = 0.03). At a median follow up of 35 months 5/122 in the BM- group and 6/61 in the BM+ group have relapsed (p = 0.2), while 1/122 and 4/61 have died of disease (p = 0.04) In conclusion, ultrastaging of breast cancer patients may identify a substantial subgroup of patients N-/BM- who may not require adjuvant chemotherapy, as well as a subgroup N-/BM+ with a decreased survival who may need more aggressive therapies. Further follow-up is needed to confirm this hypothesis, and several studies are under way.

[Hormone-dependence and C-ERBB2: relationship between estrogen receptor expression, estrogen-regulated proteins, and C-ERBB2 in breast carcinoma]. / Ormono-dipendenza e C-ERBB2: rapporto tra espressione dei recettori estrogenici, estrogen regulated proteins e C-ERBB2 nel carcinoma della mammella.

We reviewed 1491 consecutive cases of operated breast cancer between 1999-2004, and found that hormone-dependence, particularly if evaluated with functional markers of the estrogen receptor (estrogen regulated proteins, ERP), is inversely proportional to antigen neu expression. Our data confirms that ERP can give additional information on the probability of response to hormonal therapy.

[Towards the standardization of sentinel lymph node analysis in women with breast cancer]. / Verso una standardizzazione dell'analisi del linfonodo sentinella in donne con cancro della mammella.

Sentinel lymph node biopsy allows enhanced pathology through serial sections and immunohistochemical analysis of the retrieved node, with detection of micrometastases and isolated tumor cells not otherwise recognized. We present our experience with a simple, effective, pathology protocol requiring six couples of sections at three different sentinel lymph node levels. Additional micrometastases or ITC were diagnosed in 51/416 patients (14.6%).

[Does incidence of sentinel lymph node metastasis increase after diagnostic manipulation in women with breast cancer?]. / Aumenta l'incidenza di metastasi nel linfonodo sentinella dopo manipolazione diagnostica in donne con cancro della mammella?

503 patients were retrospectively evaluated to assess whether a previous needle or core biopsy, or surgical surgical excision of the primary tumor are associated with passive dislodgment of tumor cells in the sentinel lymph node, as reported in recent publications. We could not identify any increased incidence of sentinel lymph node micrometastases or isolated tumor cells after diagnostic manipulation of the primary tumor.

[Predictive variables of lymphatic metastasis in breast carcinoma with a diameter below 2 cm]. / Variabili predittive di metastasi linfonodali nei carcinomi mammari di diametro inferiore a cm 2.

The axillary lymph node status of patients with newly diagnosed breast cancer remains the most important prognostic information available at the moment. However, only a minority of patients presents with such node metastases at diagnoses. We reviewed our database and studied 500 consecutive patients with early breast cancer, and found that age inferior to 50 years, high grade, diameter superior to 1 cm, elevated Ki-67, and expression of oncogene p-53 are all factors associated with lymph node metastases.

[Extemporaneous examination of the sentinel lymph node in breast cancer: is the glass half full or half empty?]. / L'esame estemporaneo del linfonodo sentinella nel cancro della mammella: il bicchiere è mezzo pieno o mezzo vuoto?

Intra-operative examination of sentinel LN is controversial. Concordance with definitive exam of SLN in this series was 81%, though only 54% of positive cases were diagnosed. Micrometastases and ITC were usually lost intraoperatively, accounting for 14% of cases. Frozen section and touch prep of the SLN were approximately equivalent. The latter has the advantage of preserving tissue for step-analysis of SLN. The ultimate method of intraoperative analysis of SLN which can combine cost-effectiveness and accuracy needs to be determined.

Clinical applications of 2D and 3D CT imaging of the airways--a review.

Hardware and software evolution has broadened the possibilities of 2D and 3D reformatting of spiral CT and MR data set. In the study of the thorax, intrinsic benefits of volumetric CT scanning and better quality of reconstructed images offer us the possibility to apply additional rendering techniques to everyday clinical practice. Considering the large number and redundancy of possible post-processing imaging techniques that we can apply to raw CT sections data, it is necessary to precisely set a well-defined number of clinical applications of each of them, by careful evaluation of their benefits and possible pitfalls in each clinical setting. In diagnostic evaluation of pathological processes affecting the airways, a huge number of thin sections is necessary for detailed appraisal and has to be evaluated, and information must then be transferred to referring clinicians. By additional rendering it is possible to make image evaluation and data transfer easier, faster, and more effective. In the study of central airways, additional rendering can be of interest for precise evaluation of the length, morphology, and degree of stenoses. It may help in depicting exactly the locoregional extent of central tumours by better display of relations with bronchovascular interfaces and can increase CT/bronchoscopy sinergy. It may allow closer radiotherapy planning and better depiction of air collections, and, finally, it could ease panoramic evaluation of the results of dynamic or functional studies, that are made possible by increased speed of spiral scanning. When applied to the evaluation of peripheral airways, as a completion to conventional HRCT scans, High-Resolution Volumetric CT, by projection slabs applied to target areas of interest, can better depict the profusion and extension of affected bronchial segments in bronchiectasis, influence the choice of different approaches for tissue sampling by better evaluation of the relations of lung nodules with the airways, or help to detect otherwise overlooked slight pathological findings. In the exploration of the air-spaces of the head and neck, targeted multiplanar study can now be performed without additional scanning by retro-reconstructed sections from original transverse CT slices. Additional rendering can help in surgical planning, by simulation of surgical approaches, and allows better integration with functional paranasal sinuses endoscopic surgery, by endoscopic perspective rendering. Whichever application we perform, the clinical value of 2D and 3D rendering techniques lies in the possibility of overcoming perceptual difficulties and 'slice pollution', by easing more efficient data transfer without loss of information. 3D imaging should not be considered, in the large majority of cases, as a diagnostic tool: looking at reformatted images may increase diagnostic accuracy in only very few cases, but an increase in diagnostic confidence could be not negligible. The purpose of the radiologist skilled in post-processing techniques should be that of modifying patient management, by more confident diagnostic evaluation, in a small number of patients, and, in a larger number of cases, by simplifying communication with referring physicians and surgeons. We will display in detail possible clinical applications of the different 2D and 3D imaging techniques, in the study of the tracheobronchial tree, larynx, nasal cavities and paranasal sinuses by Helical CT, review relating bibliography, and briefly discuss pitfalls and perspectives of CT rendering techniques for each field.

Contralateral solitary testis metastasis antedating renal cell carcinoma: a case-report and review.

A 56-year-old Caucasian smoker male patient underwent right orchiectomy and spermatic cord resection for intrascrotal transilluminating mass. Histology findings suggested diagnosis of metastasis from Renal Cell Carcinoma (RCC). After positive CT-scan for primary neoplasm, total left nephrectomy was accomplished. Patient referred to our institution and started interferon immunotherapy; nowadays, after 21 months is disease free and in good health. We have reviewed and summarized the international literature regarding testicular metastasis from renal cell carcinoma; this is the first case of histology-proved-RCC ("clear cell" type)-solitary-contralateral-testicular-recurrence antedating clinical evidence of the primary neoplasm, which underwent orchiectomy before nephrectomy.

Neuroendocrine carcinoma of the skin: a case report of "multiple neuroendocrine carcinoma" onset and long-term remission after chemotherapy.

We herein report a case of "multiple neuroendocrine carcinoma" onset, with a torpid course, metastasis in the left breast, and an initial considerable sensitivity to the chemotherapy. We conclude that a re-evaluation of the studies on this neoplasia carried out to date is needed, in order to identify new regimens, which increase the complete response rates and their duration.

Focal ultrasound lesions in cirrhotic liver diagnosed as regenerative nodules by biopsy. A morphometric analysis.

Ten cirrhotic patients with ultrasonically discernible focal liver masses underwent fine cutting needle biopsy. Specimens were obtained from the focal lesions under ultrasound guidance and histologically diagnosed as regenerative nodules. An image analyzer was then used to determine the cytoplasmic area, nuclear area, and nuclear/cytoplasmic ratio for 100 randomly selected cells from each specimen. Data were then compared with data for specimens of normal liver tissue and data from patients with alcoholic or posthepatic cirrhosis or well-differentiated hepatocellular carcinoma (HCC). The morphometric parameters for the group of regenerative nodule specimens fell within an intermediate range between those for HCC and the nondysplastic samples, strongly suggesting a preneoplastic nature. Nine of the 10 regenerative lesions showed liver cell dysplasia, and 3 of these patients developed HCC during follow-up. Ultrasonically discernible focal masses in a cirrhotic liver should be considered preneoplastic, if not neoplastic lesions and treated aggressively to prevent their progression to outright malignancy.

[Merkel cell carcinoma of the skin. Two clinical cases]. / Carcinoma a cellule di Merkel della cute. Considerazioni su due casi clinici.

In this study are described two clinical cases of Merkel cell tumor treated in the first half of 1991. The MCC is a rare primary skin tumor with an aggressive biologic nature. The elective therapy of primary sites is surgical management with lymphadenectomy although this kind of treatment is debated. Local recurrence high incidence requires a frequent and careful follow-up with possibility of second surgical treatment in the cases without metastasis and a radiochemotherapy in the patients affected by systemic recurrences.

Hypoechoic lesions in fatty liver. Quantitative study by histomorphometry.

Ten patients with fatty liver changes were subjected to liver biopsies. In seven, ultrasonography showed focal hypoechogenicity within a "bright" liver, generally interpreted as focal sparing. Three patients had hyperechoic areas surrounded by "normal" parenchyma usually felt to represent local fat accumulation without diffuse involvement. Morphometric analysis was used to determine the total area occupied by fat (area density) and the number of lipid droplets (numerical density) per microscopic field in lesional and perilesional specimens. Compared with respective perilesional values, hypoechoic lesional numerical densities were significantly lower in 6 of 7 patients; the three hyperechoic samples had significantly higher values. In most cases, lesional and perilesional area densities were not significantly different. These hypoechoic focal lesions are believed to be merely areas in which a similar quantity of fat is contained in fewer droplets, and focal hyperechogenicity is believed to result from a larger number of fat-filled vacuoles with respect to that of the surrounding parenchyma. Variation in the number of solid-liquid interfaces causes the ultrasonic contrast between these lesions and the similarity fatty liver parenchyma surrounding them.