Endoplasmic Reticulum Chaperones Are Potential Active Factors in Thyroid Tumorigenesis.

The study aimed to evaluate the proteomic changes in benign follicular adenoma versus malignant follicular variant of papillary thyroid carcinoma. Tumor and nontumor adjacent samples were analyzed by liquid nanochromatography mass spectrometry, and protein abundance was evaluated by label-free quantification. Western blotting and quantitative real-time polymerase chain reaction were used to validate and complement the mass spectrometry data. The results demonstrated deregulated expression of four endoplasmic reticulum chaperones (78 kDa glucose-regulated protein, endoplasmin, calnexin, protein disulfide-isomerase A4), glutathione peroxidase 3 and thyroglobulin, all of them involved in thyroid hormone synthesis pathway. The altered tissue abundance of endoplasmic reticulum chaperones in thyroid cancer was correlated with serum expression levels. The identified proteins significantly discriminate between adenoma and carcinoma in both thyroid tissue and corresponding sera. Data are available via ProteomeXchange with identifier PXD004322.

THE RELEVANCE OF CYTOLOGICAL DIAGNOSTIC IN THE MAMMARY GLAND CANCER.

UNLABELLED: Breast cancer is the second leading cause of cancer death in women, while in Eastern Europe the most common form of diagnosed cancer. Out of the multiple possibilities of early detection of mammary neoplasia that have been elaborated, only mammography has proved to be a simple, efficient method and of a high sensitivity, almost 90% However, the cytological confirmation of diagnosis allows us to perform the preoperative radiotherapy treatment or poly chemotherapy. MATERIAL AND METHODS: we analyzed the informative value of these diagnosis methods in stage I mammary gland cancer (MGC). In this way, in the present paper we demonstrated that collecting samples through fine-needle aspiration biopsy allows the cytological confirmation of the diagnosis of stage I MGC in 30.7% cases. RESULTS AND DISCUSSION: In stage I MGC young patients, under 35 years, the cytological confirmation rate is 22.2% and is lower as compared to the cytological confirmation rate in patients older than 35 years which is 37.9% Also, for a tumor diameter < 0.5 cm, the prevalence of cytological confirmation was only 10.3%, while for the diameter of 0.6-1.0 cm the cytological confirmation was around 40.0%. Therefore, in order to improve the cytological diagnosis confirmation rate the tumor biopsy through the USG of the mammary glands is required. Moreover, the cytological investigation of the smear obtained by the first and second puncture was instrumental in confirming the diagnosis in 41.3% and 17.4% cases; the subsequent repetition of the punctures was not useful as it helped to confirmation of the diagnosis only in 9.3% cases. The frequency of diagnosis cytological confirmation depends on the tumor histopathological form and type of growth. CONCLUSIONS: Thus, the lowest prevalence was in the mixed forms--12.5% cases, lobular cancer--24.4% cases, while regarding the type of growth, for the rare forms the cytological confirmation rate was 7.7% and 31.5% cases for the schiros growth type.

THE IMPORTANCE OF CLINICAL AND INSTRUMENTAL DIAGNOSTIC IN THE MAMMARY GLAND CANCER.

Breast cancer is the most common oncology disease in women and is one of the major public health issues. Worldwide, is the second leading cause of cancer death in women and cancer research is a priority in all the laboratories of the world, in terms of uncovering the appearance causes of the malignant process, understanding the mechanisms of development, but most of all, the discovery of early diagnostic methods and effective treatment. Ignorance, fear of diagnosis, lack of health education and of efficient programmes for prevention and screening could cause diagnosis of the disease to be detected in the majority of cases in advanced stages, when treatment remains only palliative and very costly, in this cases the patient's suffering being immense. In this way, regarding the clinical diagnosis in stage I mammary gland cancer, in the 496 stage I MGC patients, during the primary clinical investigation the diagnosis of stage I MGC was established only in 165 (33.3%) patients, and in 232 (46,8%) patients the diagnosis of suspicion MGC was obtained. Also, in terms of instrumental diagnosis, such as mammography, ultrasonography in mammary gland cancer stage I, it seems that in accordance with literature data the pathological process features assessment in the mammary gland is problematic especially in young age. Thus, it seems that MGC represents a polymorphic and pathogenic disease and it cannot be admitted that all subgroups of patients will obtain identical results from one tactic of treatment determined for all the patients with MGC. In this way, the concept of MGC both clinical and patho morphological, combines different cell clones depending on its microstructure and biology. As a result, the evolution of the disease, the prognosis and the effectiveness of the treatment may vary in different patients at the same stage, depending on the degree of malignancy of the tumor, its histopathological structure, the degree of expression of molecular markers identification and also immune resistance.

[Clinico-biological issues of systemic lupus erythematosus patients]. / Contributii la cunoasterea clinico-biologica a lupusului eritematos sistemic si relatia cu indicatorii de activitate si distructie tisulara.

AIM: To investigate clinical and biological aspects and to evaluate potential relation with activity and damage indexes in patients with Systemic Lupus Erythematosus (SLE). MATERIAL AND METHODS: Retrospective observational study in 30 consecutive SLE patients (ACR 1997 diagnostic criteria) assessed according to a standard protocol including demographic, clinical and biological (hematology, inflammatory, immunology) data, disease activity (SLEDAI) and damage (SLICC/ACR). RESULTS: 90% SLE women with a mean age of 45.53+13.57 years and a mean age at onset of 31.10 +/- 9.20 years, presenting mainly with skin (43.33%) and small joint (56.66%) involvement, but also with renal (26.66%), cardio-vascular (26.66%) and neurological (6.66%); high anti-DNA double stranded antibodies (30% cases; mean 40.63 +/- 71.62IU/L) and low C3 levels (26.66%; mean 100.63 +/- 26.06 IU/L) have been reported, while more than 76% SLE patients displayed a low disease activity (mean SLEDAI 4 +/- 3.64) and limited damage (mean SLICC/ACR 1.4 +/- 0.85). Statistically significant correlation (p < 0.05) have been identified between SLEDAI and SLICC/ACR (r = 0.477), anti-DNA double stranded and SLEDAI (r = 0.515) respectively SLICC/ACR (r = 0.404), age at onset respectively current age and SLICC/ACR (r1 = 0.495, r2 = 0.468). CONCLUSIONS: The clinical and biological study based on data from consecutive SLE patients has offered a comprehensive approach of different disease subtypes in North-East Romania. The predominance of currently low disease activity and minimal damage disease profile could reflect either SLE particularities or the expected effects of early individualized therapy.

Knee osteoarthritis, dyslipidemia syndrome and exercise.

UNLABELLED: The aim of our study was to evaluate the influence of aerobic training on the dyslipedemia in patients with knee osteoarthritis (KOA). MATERIAL AND METHODS: Prospective observational six-month study performed on 40 patients with KOA, fulfilling the inclusion criteria, classified according to their participation in specific aerobic training program (30 minutes/day, 5 days/ week) in two subgroups. A standard evaluation protocol was followed assessing lipid parameters (total cholesterol, triglycerides, LDL-cholesterol, HDL-cholesterol levels) at baseline, three and six months. Statistical analysis was performed in SPSS 16.0, p < 0.05. RESULTS: Subgroup analysis has demonstrated a statistical significant improvement in plasma lipids levels in all patients performing regular aerobic training (cholesterol, triglycerides, HDL-cholesterol, LDL-cholesterol) (p < 0.05). Although the difference reported for total cholesterol, triglycerides and LDL-cholesterol after six months between subgroups was not significant (p > 0.05), the mean level of HDL-cholesterol was significantly higher in patients performing aerobic training, reaching the cardio-vascular protective levels. CONCLUSIONS: Regular aerobic exercise has a positive effect on plasma lipoprotein concentrations; further research is needed for the assessment of long-term effects of physical exercises for both KOA and lipid pattern.

Carotid intima-media thickness and plaque as surrogate biomarkers of atherosclerosis among consecutive women with systemic lupus erythematosus.

BACKGROUND: In recent years, there has been a growing interest in understanding the pathogenic pathways of premature accelerated atherosclerosis (AS) in systemic lupus erythematosus (SLE). However, the role of both traditional and non-traditional, SLE-specific risk factors is still under debate. AIM: To assess surrogate biomarkers of subclinical AS in SLE and to evaluate potential relations with cardiovascular risk factors. PATIENTS AND METHODS: Prospective observational study on 35 consecutive SLE women (ACR 1987 diagnostic criteria) evaluated according to a standard protocol including traditional cardiovascular risk factors (hypertension, obesity, diabetes mellitus, cigarette smoking, abnormal lipid metabolism), SLE-specific risk factors (renal disease, SLE activity and duration, corticosteroid therapy) and surrogate biomarkers of subclinical AS (carotid intima-media thickness, plaque) (B-mode color Doppler ultrasound, 7-10 MHz probe). Data were analyzed in SPSS 16 software, p<0.05. RESULTS: Significant differences (p<0.05) among subgroups (with and without plaque, thickened and normal intima) have been registered; moreover, statistical significant correlations between cIMT and age (r=0.476), age at onset (r=0.451), VLDL (r=0.382), hsCRP (r=0.436), Framingham score (r=0.421) have been reported. In addition, significant association between homocysteine and SLE-duration (r=0.460), SLEDAI (r=0.466), SLICC÷ACR (r=0.846) has been demonstrated, while hsCRP was associated with ESR (r=0.472), C3 (r=0.396), SLEDAI (r=0.569) and age (r=-0.681). Several predictors for increased cIMT have also been identified (ANOVA): hsCRP (p=0.016), VLDL (p=0.037), Framingham (p=0.012). CONCLUSIONS: Our data advocate for increased cardiovascular burden in SLE and support the value of cIMT and carotid plaque as surrogate AS biomarkers in women with SLE.

Evaluation of serum and gingival crevicular fluid C-reactive protein and IL-6 levels in patients with periodontitis and transient ischemic attacks.

BACKGROUND: Recent advances have suggested that periodontitis (PD), the paradigm of chronic infection in dental pathology, shares several pathogenic pathways with cardio- and cerebro-vascular disorders (CVD), based on inflammatory mediators including IL-1, IL-6, TNF-α. AIM: To assess pro-inflammatory biomarkers (C-reactive protein - CRP, IL-6) in serum and gingival crevicular fluid (GCF) in patients with PD and with transient ischemic attacks (TIAs). MATERIALS AND METHODS: Prospective observational study on 143 patients classified as follows: 40 healthy subjects (group A), 50 PD patients (group B) and 53 PD-TIAs patients (group C). The predefined assessment protocol has included: current medical data, risk factors for CRP changes, periodontal status (clinical, orthopantomography, Schei Ruler technique), inflammatory biomarkers (CRP, IL-6). RESULTS: High serum CRP and IL-6 have been reported in both TIAs and PD, while statistically significant increase in GCF CRP only in PD-TIAs (p<0.05). Moreover, both generalized and localized chronic PD may be at higher risk for CVD, since CRP level was higher in these subgroups. However, no significant differences were reported in serum IL-6 between generalized and localized PD. A score function was demonstrated, including bone loss degree, bleeding index, collection site depth, serum and GCF IL-6 and CRP, tooth loss, allowing the classification of PD based on risk for developing TIAs. CONCLUSIONS: CRP and IL-6 are commonly involved in the pathways of PD and TIAs. Interdisciplinary assessment should be promoted in order to implement the stratification of PD patients according to the risk for TIAs as suggested by the proposed algorithm.

Neoangiogenesis in cervical cancer: focus on CD34 assessment.

UNLABELLED: Despite recent advances in understanding the immune mechanisms of cervical cancer (CC), relapse remains still an actual issue and recognition of new predictive biomarkers is essential. AIM: The purpose of this retrospective study was to investigate neo-angiogenesis in CC and its possible utility as prognostic biomarker. MATERIAL AND METHODS: Paraffin-embedded tissue samples from 61 consecutive women with CC were immunostained for CD34 and E-cadherin. Statistical analysis was performed in SPSS-12 software, p<0.05. RESULTS: Statistically significant differences between CD34 distribution among three interest tumor regions: micro-vessels density increase from central to peripheral area (chi(2), p<0.05); statistically significant correlation between CD34 expression, particularly in stromal and peripheral sites, E-cadherin (Spearman r1=-0.321) and lymphatic invasion (Spearman r2=0.455) (p<0.05) were reported. Overall five-year survival is clearly dependent on level and distribution of tumor angiogenesis among defined area of interest as suggested by Kaplan-Meier analysis. CONCLUSIONS: Angiogenesis is essential for guiding CC evolution and prognosis, particularly in squamous invasive types.

Predictive value of cellular immune response in cervical cancer.

UNLABELLED: Despite recent advances in the immune mechanisms of cervical cancer (CC), the relapse still remains an actual issue and recognition of new predictive biomarkers is essential. AIM: The purpose of this retrospective study was to investigate possible differences in the primary, in situ, cellular immune response between cervical carcinoma with and without relapse. MATERIAL AND METHODS: Paraffin-embedded tissue samples from 61 consecutive women with CC (34 with and 27 without relapse) were immunostained for CD3, CD20 and CD45 cells. Immune cell profile densities were further assessed, assigning scores between 0 and 3: "0" meaning the absence of inflammatory infiltrate, "1+" low, "2+" intense and "3+" intense infiltrate with lymphoid follicles. Statistical analysis was performed in SPSS-13 software, p<0.05. RESULTS: Statistically significant intra- and peri-tumoral low numbers of several immune cell subtypes are strongly associated with relapse of disease within three and five years in patients with CC (p<0.05); moreover, statistical significant correlations between immune cells and both free survival (CD3: r=0.382; CD20: r=0.404; CD45: r=0.376) and relapse (CD3: r=-0.408; CD20: r=-0.355; CD45: r=-0.354) have been demonstrated. Only CD3 was reported as predictive biomarker of relapse in CC (ANOVA, t-Student, p<0.05). CONCLUSIONS: Major differences in the cellular immune response among patients with cervical cancer with and without relapse within three and five years have been demonstrated. CD3 may be used as potential prognostic biomarkers, whereas the results are promising for adjuvant immunotherapy.

Tumor biomarkers in cervical cancer: focus on Ki-67 proliferation factor and E-cadherin expression.

UNLABELLED: Despite recent advances in the immune mechanisms of cervical cancer (CC) and complex management opportunities, relapse remains still an actual issue. While predictive factors are required, current research is directed towards proliferation and tumor aggressiveness biomarkers as potential negative factors in CC. The main objectives were to assess tumor proliferation and invasiveness biomarkers (Ki-67, E-cadherin) and to identify potential correlation between biomarkers and classic prognostic factors in CC. Radical hysterectomy specimens from 61 consecutive CC were immunohistochemically investigated for Ki-67 and E-cadherin. Nuclear immunostaining for Ki-67 proliferation index was assigned scores 1 to 3, "+" meaning low (10-30%), "++" moderate (30-50%), "+++" high-proliferation rate (>50%); cell membrane E-cadherin staining was either negative or positive. Statistical analysis was performed in SPSS-13 software, p<0.05. RESULTS: no significant correlation between Ki-67 and classical prognostic factors (p>0.05) was reported; however, in relapsed CC, Ki-67 correlates with tumor grading (r=0.386, p<0.05). Significant correlation between E-cadherin and tumor size (r=-0.280, p=0.029), relapse (r=-0.386, p=0.002) and disease free survival (r=0.374, p=0.003) were demonstrated. Indirect statistically significant moderate correlation between Ki-67 and E-cadherin (r=-0.461, p<0.00001) was shown, mainly in invasive squamous CC (r=-0.549, p=0.0001), stage IB (r=-0.578, p=0.009), IIB (r=-0.585, p=0.003), relapsed CC (r=-0.525, p<0.01), HPV-infection (r=-0.504, p=0.033). CONCLUSIONS: CC aggressiveness, particularly in invasive squamous carcinoma, either 16 or 18 HPV-positive cases, FIGO stage IB and IIB, and cases with relapse, depends on two pivotal factors, tumor proliferation rate (Ki-67) and tumor invasiveness (E-cadherin).

Immunohistochemical study of skeletal muscle in rheumatoid myositis.

INTRODUCTION: Rheumatoid myositis (RM) represents a poorly characterized entity, immune mechanism, assessment and management remaining still unclear. The aim of this study was to investigate endothelial and inflammatory cells activation in RM muscle biopsy. MATERIAL AND METHODS: Prospective study on 23 consecutive rheumatoid arthritis (RA) with muscle involvement as defined by clinical, biological and imagistic parameters. CD4, CD8, CD20, CD3, CD45RO and CD68 markers, HLA-DR, cytokines receptors (IL-2, TNFalpha, TGF alpha), pro-apoptotic (CD95) and adhesion molecules (CD54) were assessed by immunohistochemistry in deltoid muscle samples. RESULTS: (1) endomysial, perivascular and perimysial inflammatory infiltrates and moderate muscle fibers involvement; predominance of activated (HLA-DR+), memory (CD45RO+) CD3+TCD8+ cells and macrophages surrounding and invading non-necrotic muscle fibers (34.78%) and TCD4+ activated cells in perivascular and perifascicular areas (65.22%); (2) up-regulation of HLA-DR, CD54 and IL-2R on both endothelial cells and lymphocytes (85%); (3) aberrant increased CD95 in endothelial cells without any other apoptotic sign (83%) have been described. CONCLUSION: Increased expression of activation markers, adhesion molecules and cytokine receptors may indicate early endothelial activation in RM pathogenesis, while endomysial TCD8+ activation may account for further development and perpetuation of myositis.

Adalimumab therapy in patients with active rheumatoid arthritis.

AIM: To evaluate efficacy and safety of adalimumab (ADA), a monoclonal anti-TNFalpha antibody, in rheumatoid arthritis (RA). MATERIAL AND METHOD: 5 years retrospective observational study on 70 active RA (ARA 1987 modified criteria; 48 women; mean age 52.6 +/- 11.7 years; mean disease duration 6.7 +/- 3.2 years, mean DAS28 6.5 +/- 1.3) treated with ADA (classic regimen). All patients have been assessed according to a standard protocol: (i) clinical (tender and swollen joints; pain; global disease evaluation), (ii) inflammatory and (iii) immune parameters (total antinuclear and anti-double stranded DNA antibodies), (iv) activity and functional scores, (v) response to therapy (EULAR), (vi) adverse events. Evaluation was performed at baseline and every 3 months. Statistical analysis was done in SPSS-13, p < 0.05. RESULTS: Statistical significant improve in RA activity (mean final DAS28 3.6 +/- 0.8, p < 0.05), functional scores (mean HAQ 1.3 +/- 0.3, p < 0.05) and decreased X-ray progression (Sharp score) have been reported; 60% RA were responders (mean EULAR 2.7 +/- 1.2), 35.7% in remission, while switching to another biological agent (14.28% ADA failure) was done in 20% cases, clinical, biological and radiological efficacy and favorable safety profile of ADA have been demonstrated in real life long-term administration in active RA.