Long-term cadmium exposure influences the abundance of proteins that impact the cell wall structure in Medicago sativa stems.

Cadmium (Cd) is a non-essential, toxic heavy metal that poses serious threats to both ecosystems and human health. Plants employ various cellular and molecular mechanisms to minimise the impact of Cd toxicity and cell walls function as a defensive barrier during Cd exposure. In this study, we adopted a quantitative gel-based proteomic approach (two-dimensional difference gel electrophoresis) to investigate changes in the abundance of cell wall and soluble proteins in stems of Medicago sativa L. upon long-term exposure to Cd (10 mg·Cd·kg-1 soil as CdSO4 ). Obtained protein data were complemented with targeted gene expression analyses. Plants were affected by Cd exposure at an early growth stage but seemed to recover at a more mature stage as no difference in biomass was observed. The accumulation of Cd was highest in roots followed by stems and leaves. Quantitative proteomics revealed a changed abundance for 179 cell wall proteins and 30 proteins in the soluble fraction upon long-term Cd exposure. These proteins are involved in cell wall remodelling, defence response, carbohydrate metabolism and promotion of the lignification process. The data indicate that Cd exposure alters the cell wall proteome and underline the role of cell wall proteins in defence against Cd stress. The identified proteins are linked to alterations in cell wall structure and lignification process in stems of M. sativa, underpinning the function of the cell wall as an effective barrier against Cd stress.

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.

Treatment of prurigo nodularis with pregabalin.

WHAT IS KNOWN AND OBJECTIVE: Prurigo nodularis (PN) is a chronic skin condition that is difficult to treat. Pregabalin is one of the possible treatments for PN but its safety and efficacy are not well defined. We aimed to assess the efficacy of pregabalin in patients with PN. METHODS: Thirty patients (10 men, 20 women; mean age 51.6 ± 9.39 years) were treated with pregabalin (75 mg/day) for 3 months. Efficacy was classified as (i) successful (disappearance of the pruritus and reduction of nodules); (ii) slight improvement/reduction of the nodules, that is, number and/or flattening, no disappearance of itching; or (iii) unsuccessful. RESULTS: Twenty-three patients (76%) responded successfully after 3 months of treatment. There was a statistically significant difference between visual analogue scale scores before and after 1 month treatment period (8·15 ± 2·04 and 1·5 ± 1·12, respectively; P < 0·0001). Pregabalin was generally well tolerated with only six (20%) patients reporting side effects. No patient showed any renal insufficiency. WHAT IS NEW AND CONCLUSION: In our study, pregabalin was effective for the treatment of PN. However, given the open and non-controlled study design used, a properly powered randomized controlled validation study is called for.

COX-2 expression in lobular in situ neoplasia of the breast: correlation with histopathological grading system according to the Tavassoli classification.

AIMS: There is considerable evidence to link cyclooxygenase (COX)-2 to the development of cancer. The aim of this study was to assess COX-2 expression and its subcellular localization in lobular in situ neoplasia (LIN) of the breast and to verify differences in COX-2 expression between different grades of lesions according to the Tavassoli classification. METHODS AND RESULTS: We analysed the expression of COX-2 protein by immunohistochemistry in tissue samples of 51 LIN lesions classified into three grades according to the Tavassoli classification. COX-2 immunostaining was observed in 78.4% of LIN samples and showed a prevalent membranous rather than cytoplasmic pattern. COX-2 was expressed in 16/17 (94.1%) LIN1, 22/25 (88%) LIN2 and 2/9 (22.2%) LIN3. As regards COX-2 expression, a statistically significant difference was found between LIN1 and LIN3 (P = 0.001) and between LIN2 and LIN3 (P =0.001). No difference was found between LIN1 and LIN2. Moreover, a significant negative correlation was found between LIN grade and COX-2 expression (P < 0.0001). CONCLUSIONS: COX-2 is highly expressed in LIN, supporting a role for this protein in the early stage of breast carcinogenesis, representing the rationale for using COX-2 selective inhibitors in the earliest stages of breast cancer.

Neurodistribution of androgen receptor immunoreactivity in the male frog, Rana esculenta.

Sexual behavior in vertebrates depends on the cyclic release of steroids and their binding to the brain receptors. Previously, we demonstrated the presence of specific binding of (3)H-testosterone and staining with PG-21 in the brain of the adult male frog, Rana esculenta. Here, we report our further receptor characterization using an anti-androgen receptor antiserum, PG-21, and the androgen site of action in frog brain. Nuclei, which contained cells labeled for the androgen receptor (AR), were mainly identified in the olfactory bulbs, preoptic-septal region, infundibulum, amygdala, thalamus, tectum, torus semicircularis, and medulla. The neuroanatomical AR staining appears similar to that in other lower vertebrates.

Vitamin E in early stages of sea bass (Dicentrarchus labrax) development.

This study reports titration of vitamin E levels in the sea bass (Dicentrarchus labrax) using high-pressure liquid chromatography. The first part of the work is devoted to vitamin E detection in: (1) plasma of maturing females and males characterized by different body sizes; (2) seminal fluid and eggs; and (3) developing embryos of sea bass fed with vitamin E. In the second part of the study, variations of vitamin E levels during larval development are analyzed. The results show a direct correlation between plasma vitamin E content and body size for both adult male and female sea bass. High vitamin E levels were found in seminal fluid, in eggs before and after fertilization, and in embryos during development and at hatching, whereas vitamin E level was low in dead embryos and in embryos with limited survival. During larval development, the vitamin E content decreased slowly but steadily during the first four days of larval growth; subsequently, it progressively increased from day 9 to day 40. In teratogenic larvae, vitamin E content was significantly higher than in normal larvae. This study provides evidence on how vitamin E exerts an antioxidant defense in sea bass reproduction.

['Incidentaloma' in inguinal hernia surgery]. / 'Incidentaloma' nella chirurgia dell'ernia inguinale.

Adult Granulosa-Cell Tumor (GCT) is an uncommon ovarian malignancy. The clinical course is characterized by slow growth with late recurrence. A variety of clinical and pathologic parameters have previously been evaluated for prognosis with inconclusive results. The aim of this study was to report a case of a granulosa-cell tumor of the ovary that recurred 25 years after first surgery. The clinical course and histopathology of the case were reviewed and a literature search for other reported cases was performed.

[Sigma's stenosis. Clinical, diagnostic and therapeutical problems]. / Le stenosi del sigma. Problematiche cliniche, di diagnostica e terapeutiche.

Sigma's stenosis represent a frequent pathology, but its very difficult to have a pre-operatory diagnosis about the nature of these stenosis, not withstanding the introduction of more sophisticated instruments of diagnosis. In this work we present 9 cases of sigma stenosis; some of these with a common etiology and other very interesting for their rare and unusual origins. The patients were underwent to a different timing of diagnosis and therapy based on clinical history and diagnostic hypotheses; our experiences demonstrate that the clinical intuition is really useful for a correct diagnosis in the patients with a sigma's stenosis and consequently, in order to plan the correct therapy.

[Multinodular goiter as an organ disease: study of continual nodulogenesis. Biological bases indicating total thyroidectomy]. / Il gozzo multinodulare come malattia d'organo: studio della nodulogenesi incessante. Basi biologiche per l'indicazione alla tiroidectomia totale.

Multinodular goitre is a very common pathology among people especially female adult and represents more than 90% of the thyroid surgery. We analyzed 37 patients, demonstrating that the whole gland is progressively affected by an incessant nodulogenesis and we confirm the necessity of total thyroidectomy in order to eliminate any persistent pathology.

CD44v6 and Nm23-H1 protein expression related to clinico pathological parameters in colorectal cancer.

OBJECTIVE: Nm23-H1 and CD44v6 expression has been shown to be correlated with the metastatic potential of colorectal cancer (CRC) in some studies but not in others. The present study was undertaken to evaluate immunohistochemically the expression of these markers and to correlate them with clinicopathological variables. MATERIALS AND METHODS: Archival tissues of 41 non metastatic colorectal cancers were histopathologically evaluated and stained with monoclonal antibodies versus Nm23-H1 and CD44v6. RESULTS: Expression of Nm23-H1 was detected in 73% (n = 30) of all CRC, and CD44v6 in 37% (n = 15) of all CRC. CD44v6 was found to be statistically associated with tumour grading differentiation (p < 0.03), but no correlation emerged between Nm23-H1 and CD44v6 and Dukes stage, site, peritumoral lymphocytic infiltration, venous infiltrating, perineural infiltrating, tumour budding, pushing and infiltrating tumour growth. CONCLUSION: Even if the results are not statistically significant, the authors noticed that the expression of Nm23-H1 was correlated with those histopathological parameters that indicate local disease progression and metastases.

[Hyperhomocysteinemia and thrombosis: clinical case and literature's review]. / Iperomocisteinemia e trombosi: caso clinico e revisione della letteratura.

Patients with homocystinuria have an increased risk for both venous and artherial thrombosis but it has been found that even moderate increase in homocysteine levels are associated with heightened risk of thromboembolism. The Authors report a case of juvenile vasculopathy in a patient with hyperomocysteinemia. In this case the patient presented both venous and artherial thromboembolism in the absence of the most important risk factors. The opportunity is stressed to perform clinical tests for the correct diagnosis of hyperhomocysteinemia because it is possible, with the use of folic acid and vitamin B12, to correct the elevated levels of homocysteine.

[Breast carcinoma with neuroendocrine differentiation. Clinical case and review of the literature]. / Il carcinoma mammario con differenziazione neuroendocrina. Osservazione clinica e revisione della letteratura.

Female breast carcinoma with neuroendocrine differentiation is a lesion which can enhance of conventional therapy added with tamoxifene-octreotide association. The Authors report a case completely responsive after two years therapy, which grading was: pT4, N1biii, Mx, G2 staging III.

Progesterone receptor: some viewpoints on hypothalamic seasonal fluctuations in a lower vertebrate.

Steroids secreted by the ovary, specifically estrogen and progesterone, influence the expression of behaviors associated with reproduction by interacting with a specific binding protein, or receptor, located in target cells in certain hypothalamic nuclei. The present paper reviews the progesterone receptor studies in the vertebrates brain, the progesterone receptor fluctuations throughout the reproductive cycle and suggests a role for progesterone receptors in the regulation of hypothalamic functions in amphibians. Furthermore, we report here a combined biochemical and immunohistochemical analysis of the hypothalamic progesterone receptor during the reproductive cycle of a lower vertebrate, the female amphibian anura Rana esculenta. 3H-Progesterone binding activity was found in both cytosol and nuclear extract samples. The progesterone binding moiety showed typical characteristics of a true receptor, such as high affinity, low capacity and specificity for progesterone. Further characterization was performed by using monoclonal antiserum raised against both the subunits A and B of the chicken progesterone receptor. Immunostained neurons were located mainly in two specific regions of the hypothalamus: the preoptic area and the infundibular hypothalamus. An immunoreactive band of about 67 kDa was observed using Western blotting, both in the cytosol and in the nuclear extract. Progesterone receptor levels fluctuated throughout the cycle along with plasma steroids and vitellogenin synthesis.

Estrogen receptors and aromatase activity in the hypothalamus of the female frog, Rana esculenta. Fluctuations throughout the reproductive cycle.

It is well known that certain actions of androgen are mediated through in situ aromatization to estrogen in neural target tissues. This study was undertaken to investigate androgen utilization in the hypothalamus of the female frog, Rana esculenta, through a quantification of estrogen receptors and aromatase activity during the reproductive cycle. 3H-estradiol-binding molecules were present in both the cytosol and the nuclear extract of the hypothalamus. These molecules bound specifically 3H-estradiol with high affinity (Kd 10(-10) M) and low capacity (cytosol: 1.2+/-0.4 fmol/mg protein; nuclear extract: 7.9+/-0.6 fmol/mg protein). Aromatase activity was detected in the microsomal fraction of the hypothalamus using a sensitive in vitro radiometric assay. Both aromatase activity and nuclear estrogen receptor binding fluctuated in synchrony throughout the reproductive cycle. Western blot analysis of aromatase protein revealed one immunoreactive band with a molecular weight of approximately 56 kDa. In contrast to aromatase enzyme activity, the relative levels of aromatase protein changed little during the reproductive cycle suggesting that post-translational mechanisms may be involved in regulating estrogen synthesis in the frog brain. A possible role for estrogens in the modulation of the reproductive behavior in this species is suggested.

Cisplatin, vindesine, mitomycin-C and 13-cis retinoic acid in the treatment of advanced non small cell lung cancer. A phase II pilot study.

BACKGROUND: Thirteen-cis retinoic acid (RA) has been shown to have growth-inhibitory and differentiative activity on non-small cell lung cancer (NSCLC) cells in vitro. This promoted the rationale for combining RA with three active drugs, cisplatin (CDDP) vindesine (VDS) and mitomycin-c (MMC) in the treatment of advanced NSCLC. PATIENTS AND METHODS: Patients with a histologically confirmed non-resectable NSCLC, measurable lesion, performance status < or = 3, and informed consent were enrolled. The chemotherapy schedule included cisplatin 60 mg/m2 and mitomycin-c 10 mg/m2 day 1 and vindesine 3 mg/m2 on days 1 and 5, every 4 weeks. RA was administered orally, at a dose of 0.5 mg/kg, 5 days per week, during chemotherapeutic intervals and to responding patients until disease progression was observed. RESULTS: Thirty patients, receiving a total of 163 chemotherapy courses, were evaluated for response and toxicity. Objective responses included complete response in 2 patients (7%), partial response in 10 patients (33%), stable disease in 9 patients (30%) and progressive disease in 9 patients (30%), (response rate 40%: Confidence interval 95% 22.7% to 59.4%). Median time to progression was 8.6 months (range 3.9-45+). Median overall survival was 11.3 months (range 1-45+). The 1-year survival rate was 47%. Toxicity (WHO) included nausea and vomiting grade 2 in 6 patients, transient ileus in 3 patients and grade 3-4 leukopenia in 5 patients. Two patients underwent surgical resection of residual disease and remain in CR. CONCLUSIONS: The addition of RA to cisplatin, vindesine and mitomycin-c is feasible and shows some activity in the treatment of NSCLC, with manageable toxicity.

Effect of testosterone and 17beta-estradiol treatment on sex steroid binding proteins in the male of the green frog Rana esculenta.

In this paper we report the effect of gonadectomy and/or long-term sex steroid (testosterone and estradiol-17beta) treatment and prolonged captivity (two months) on testosterone and estradiol-17beta binding proteins (TBP and EBP, respectively) in the plasma of the male of the green frog Rana esculenta. Experiments were carried out during different periods of the reproductive cycle. Gonadectomy and prolonged captivity were carried out in winter, when the spermatogenic activity slowed down and the concentration of circulating androgens was high. Both gonadectomy and prolonged captivity resulted in a significant decrease in TBP binding activity, which could not be restored by the hormonal treatment. On the contrary, when the hormonal treatment was carried out in the early summer, when the spermatogenesis was active but the concentration of circulating androgens was low, a significant increase in TBP binding activity was observed. Neither gonadectomy, nor the prolonged captivity, nor the hormonal treatment affected EBP levels. Our data indicate that TBP apparent changes in response to testosterone and estradiol-17beta treatment varied according to the period of the reproductive cycle, an indication that studies on sex steroid binding proteins regulation should take into consideration the internal endocrine condition before drawing any final conclusion especially in species with a seasonal mode of reproduction.

Evidence of a progesterone receptor in the liver of the green frog Rana esculenta and its down-regulation by 17 beta estradiol and progesterone.

Progesterone is a versatile hormone showing an ample variety of effects. One of the numerous functions attributed to progesterone is the modulation of vitellogenesis in oviparous vertebrates. As a prerequisite for the possible involvement of progesterone in vitellogenesis modulation, we investigated the presence of a progesterone receptor (PR) in the liver of the female green frog Rana esculenta. 3H-Progesterone (3H-P) binding activity was found in both cytosol and nuclear extract of the liver of Rana esculenta. The progesterone-binding moiety showed the typical characteristics of a true receptor, such as high affinity, low capacity, and specificity for progesterone. It also bound to DNA-cellulose and was eluted with a linear salt gradient at a concentration of 0.05 M of NaCl. The progesterone-binding moiety was down regulated by steroid hormones, in that ovariectomy resulted in a significant increase, in both cytosol and nuclear extract, of 3H-P binding activity with respect to intact females. On the contrary, 3H-P binding activity was almost undetectable after estradiol and/or progesterone treatment. The progesterone binding moiety of Rana esculenta was analyzed by Western blotting with the aid of a monoclonal antibody raised against the subunits A and B of the chicken PR. An immunoreactive band of about 67 kDa was observed in the liver of both intact and treated females. The 67 kDa band showed an increased intensity in ovariectomized animals, while it was faint following treatment with estradiol and/or progesterone. This is the first report on the presence of a progesterone receptor (PR) in the liver of an amphibian. PR of Rana esculenta is down regulated by estradiol and/or progesterone and shows peculiar immunological and biochemical characteristics, which make it rather different from the PR of other vertebrates.

Ifosfamide, vinorelbine and gemcitabine in advanced non-small cell lung cancer. A phase I study.

The objective of this phase I study was to identify the maximum tolerated dose (MTD) and toxicity of a three drug, platinum-free regimen, including gemcitabine, ifosfamide and vinorelbine, in the treatment of patients with advanced non-small cell lung cancer (NSCLC). 33 chemotherapy-naïve patients with histologically confirmed, unresectable NSCLC, received fixed doses of ifosfamide (1500 mg/m2 days 1-3 with mesna) and vinorelbine (25 mg/m2 days 3 and 8). The gemcitabine dose was escalated from 500 to 1200 mg/m2 on days 3 and 8 every third week. The escalation was stopped at dose level 4 (gemcitabine 1200 mg/m2) since all 3 patients of this cohort showed dose-limiting thrombocytopenia and/or neutropenia at treatment cycle 1. The dose recommended for phase II trials is: gemcitabine 1000 mg/m2 and vinorelbine 25 mg/m2 given on days 3 and 8 plus ifosfamide 1500 mg/m2 on days 1-3. An encouraging response rate of 50% (95% confidence interval (CI): 32-68%) was observed in 32 patients evaluated. Our results show that ifosfamide, vinorelbine and gemcitabine can be safely administered as outpatient chemotherapy for NSCLC. Myelosuppression is the dose-limiting toxicity (DLT) of this regimen with no major subjective side-effects observed.