An Autoimmune Haemolytic Anaemia Secondary to Ipilimumab Treatment.

BACKGROUND: Melanoma is one of the fastest growing neoplasms worldwide. Treatment of metastatic disease has swiftly shifted in the last decade from generally ineffective chemotherapy regimens to highly effective targeted treatments or immunotherapy, with a range of side effects that differ completely from those of previous treatments for this disease. CASE: We present a case of a 71-year-old man with diagnosis metastatic melanoma. This patient was treated with anti-CTLA-4 antibody ipilimumab. Despite minor skin toxicity, the regimen was well tolerated until he developed hemolytic anemia, an autoimmune side effect of ipilimumab. The treatment was withdrawn and steroids were administered until the issue was resolved. CONCLUSION: Immunotherapy has become the standard of care for many tumors, and its side effects are completely different from those of chemotherapy, meaning that oncologists must be aware of this to avoid a potentially life-threatening situation and arrive at an early diagnosis and implement prompt treatment.Key words: anemia - autoimmune event - anti-CTLA-4.

Human periodontal ligament stem cells cultured onto cortico-cancellous scaffold drive bone regenerative process.

The purpose of this work was to test, in vitro and in vivo, a new tissue-engineered construct constituted by porcine cortico-cancellous scaffold (Osteobiol Dual Block) (DB) and xeno-free ex vivo culture of human Periodontal Ligament Stem Cells (hPDLSCs). hPDLSCs cultured in xeno-free media formulation preserved the stem cells' morphological features, the expression of stemness and pluripotency markers, and their ability to differentiate into mesenchymal lineage. Transmission electron microscopy analysis suggested that after one week of culture, both noninduced and osteogenic differentiation induced cells joined and grew on DB secreting extracellular matrix (ECM) that in osteogenic induced samples was hierarchically assembled in fibrils. Quantitative RT-PCR (qRT-PCR) showed the upregulation of key genes involved in the bone differentiation pathway in both differentiated and undifferentiated hPDLSCs cultured with DB (hPDLSCs/DB). Functional studies revealed a significant increased response of calcium transients in the presence of DB, both in undifferentiated and differentiated cells stimulated with calcitonin and parathormone, suggesting that the biomaterial could drive the osteogenic differentiation process of hPDLSCs. These data were confirmed by the increase of gene expression of L-type voltage-dependent Ca2+ (VDCCL), subunits α1C and α2D1 in undifferentiated cells in the presence of DB. In vivo implantation of the hPDLSCs/DB living construct in the mouse calvaria evidenced a precocious osteointegration and vascularisation process. Our results suggest consideration of DB as a biocompatible, osteoinductive and osteoconductive biomaterial, making it a promising tool to regulate cell activities in biological environments and for a potential use in the development of new custom-made tissue engineering.

Sentinel Lymph Node in Melanoma -  a Study Conducted in the South of Brazil.

BACKGROUND: The presence or absence of lymph node metastases is a very important prognostic factor for survival and recurrence in patients with cutaneous melanoma. Controversies remain among specialists about whether it is advisable or not to perform a sentinel lymph node biopsy in patients with melanoma, although it is currently supported by most standard guidelines. We performed this study to identify which patients are more prone to having a positive lymph node test result in a population with high melanoma rates in the south of Brazil. MATERIALS AND METHODS: We performed the study with 62 consecutive patients who underwent sentinel lymph node biopsies from 2003 to the early months of 2015 in the city of Blumenau - Santa Catarina, Brazil. RESULTS: Breslow thickness, ulceration, nodular subtype, and Clark level IV were associated with positive sentinel lymph node group status (p 0.05). DISCUSSION: Although there is still a controversy over whether or not this procedure should be performed, most guidelines still support its application. To lower the economical and physical impact, however, it is worthwhile to determine which patients are more prone to acquiring positive lymph node at presentation and, perhaps, in the future, indicate surgery for this particular group, who could benefit most from further treatment options. KEY WORDS: melanoma - sentinel lymph node biopsy - prognosis.

Forbidden to Drive -  a New Chemotherapy Side Effect.

BACKGROUND: Capecitabine is an oral antineoplastic agent classified as a pyrimidin analogue. It is widely used in different types of cancers. Unfortunately, it does not come without a cost, as the drug may have adverse effects - largely diarrhea, but also hand -foot syndrome and loss of fingerprints in extreme cases. CASE: We report a case of a 47-year-old male, which had been diagnosed with metastatic rectal cancer and received capecitabine, developing a severe hand foot syndrome which led him to lose his fingerprints in spite of complete resolution of the syndrome after stoppage of the drug. CONCLUSION: This case highlights a rare condition that, even though not precisely a disease per se, may hinder patients quality of life and must be recognised by the treating physician as an treatment related side effect.

A rare neoplastic growth on the ear lobe.

We report a case of an 83-year-old previously healthy female patient presenting with a swiftly evolving erythematous violaceous, infiltrative, ulcerated onion like mass with hyperkeratotic surface on the left ear lobe. The lesion was excised and resulted as an atypical fibroxanthoma, an extremely rare neoplastic growth, being a superficial variant of pleomorphic malignant fibrous histiocytoma. A brief review of dia-gnosis, treatment and prognosis is discussed.

Analysis of disease-free survival and overall survival in patients with luminal a breast cancer stratified according to TNM.

BACKGROUND AND AIM: The TNM classification of malignant tumours is the most commonly used system to assess the stage as well as the prognosis of cancer. However, one of the biggest challenges in treatment of breast cancer is the understanding of tumour heterogeneity typical of these carcinomas. The aim of this study was to analyse the disease-free survival and overall survival in patients with luminal A subtype of breast cancer, stratified by TNM staging system. METHODS AND STUDY DESIGN: A total of 363 medical records from January 2001 to May 2006 were evaluated for data collection. There were 136 patients with luminal A breast cancer, selected for the cohort. The main objective was the analysis of disease-free survival (DFS) and overall survival (OS) in patients with luminal A breast cancer, stratified according to the TNM classification. RESULTS: The group of 136 patients with immunohistochemically defined luminal A subtype represented 53% of the 253 patients with breast cancer. There was no significant difference in the number and type of patients as for TNM stage and histological grading among patients treated with chemotherapy and HT in comparison to patients treated with HT alone. CONCLUSION: Our analysis did not prove any significant difference in survival of patients treated with chemotherapy followed by hormone therapy in comparison to patients treated with HT alone. We suggest that the IHC luminal A subtype of breast cancer generally warrants a good prognosis independently on other prognostic factors such as TNM stage. We conclude that patients might not benefit of adding a chemotherapy to hormonal therapy in adjuvant settings.

Bevacizumab as secondline treatment of glioblastoma -  worth the effort?

OBJECTIVE: To evaluate the role of bevacizumab and irinotecan as secondline treatment of glioblastoma in patients with progression after radiotherapy and temozolomide. METHODS: A retrospective analysis of 16 subjects was performed with overall survival and toxicity evaluation as the primary endpoint. RESULTS: The analysis revealed serious toxicity of this highly expensive regimen without proving an improvement in overall survival of patients in comparison to a control group. CONCLUSION: Unless there are robust data from phase III clinical trials, including quality of life assessments or evaluation of predictive bio-markers to guide therapy, bevacizumab and irinotecan regimen should be spared for cautiously selected patients, especially in countries with limited budget for oncological treatment.

Wage -specific assessment of mammography screening in Brazilian women.

OBJECTIVE: Mammography is an inexpensive examination that has become a standard screening method thanks to its cost-effectiveness. Due to an enormous cost escalation of cancer treatment over the last years, the assessment of this method, especially in the settings of an emerging country, is a matter of serious concern. METHODS: A search within the national health database was conducted registering the data with nation- wide coverage. Data collection and analysis was commenced in 2010 with referrence to the year 2008 as the most recently updated period. The obtained information on mammography screening among Brazilian women was evaluated by subdividing the data in four groups according to socioeconomic status of the subjects. The study population was estimated to comprise about 2 million individuals. RESULTS: More than 45% of Brazilian women older than 40 years of age have never undergone a mammography. The figures correlated with the income level, reaching an astonishing number of 62.2% in a group of patients with the lowest income level. CONCLUSION: Mammography has proved effective in early detection of breast cancer in women. This study suggests that the lack of compliance in breast cancer screening in Brazil, pronounced especially in low- income families, may contribute to the increasing mortality rates associated with the disease.

Adjuvant! Online: mind the gap!

The objective of this study was to compare the actual outcome and prognosis estimated by the program Adjuvant! Online for breast cancer in Itajaí (Brazil). It is a retrospective cohort study, post-hoc analysis, in which 214 patients of three institutions were compared in real overall survival (OS) and disease-free survival (DFS) during a 3-year follow-up, and estimated OS and DFS with the program for 10 years by Adjuvant! Online, using the following variables: age, clinical stage, histological grade, lymph node involvement, immunohistochemical subtype and type of adjuvant therapy. The DFS and OS rates in the sample observed and estimated by the program demonstrated correspondence, with the curves trailing beneath the same pattern, due to shorter "real" follow-up (3 versus 10 years). The only group that demonstrated a marked decline in DFS in relation to the real prognosis was that of patients younger than 40 years.

IP3R2 levels dictate the apoptotic sensitivity of diffuse large B-cell lymphoma cells to an IP3R-derived peptide targeting the BH4 domain of Bcl-2.

Disrupting inositol 1,4,5-trisphosphate (IP3) receptor (IP3R)/B-cell lymphoma 2 (Bcl-2) complexes using a cell-permeable peptide (stabilized TAT-fused IP3R-derived peptide (TAT-IDP(S))) that selectively targets the BH4 domain of Bcl-2 but not that of B-cell lymphoma 2-extra large (Bcl-Xl) potentiated pro-apoptotic Ca(2+) signaling in chronic lymphocytic leukemia cells. However, the molecular mechanisms rendering cancer cells but not normal cells particularly sensitive to disrupting IP3R/Bcl-2 complexes are poorly understood. Therefore, we studied the effect of TAT-IDP(S) in a more heterogeneous Bcl-2-dependent cancer model using a set of 'primed to death' diffuse large B-cell lymphoma (DL-BCL) cell lines containing elevated Bcl-2 levels. We discovered a large heterogeneity in the apoptotic responses of these cells to TAT-IDP(S) with SU-DHL-4 being most sensitive and OCI-LY-1 being most resistant. This sensitivity strongly correlated with the ability of TAT-IDP(S) to promote IP3R-mediated Ca(2+) release. Although total IP3R-expression levels were very similar among SU-DHL-4 and OCI-LY-1, we discovered that the IP3R2-protein level was the highest for SU-DHL-4 and the lowest for OCI-LY-1. Strikingly, TAT-IDP(S)-induced Ca(2+) rise and apoptosis in the different DL-BCL cell lines strongly correlated with their IP3R2-protein level, but not with IP3R1-, IP3R3- or total IP3R-expression levels. Inhibiting or knocking down IP3R2 activity in SU-DHL-4-reduced TAT-IDP(S)-induced apoptosis, which is compatible with its ability to dissociate Bcl-2 from IP3R2 and to promote IP3-induced pro-apoptotic Ca(2+) signaling. Thus, certain chronically activated B-cell lymphoma cells are addicted to high Bcl-2 levels for their survival not only to neutralize pro-apoptotic Bcl-2-family members but also to suppress IP3R hyperactivity. In particular, cancer cells expressing high levels of IP3R2 are addicted to IP3R/Bcl-2 complex formation and disruption of these complexes using peptide tools results in pro-apoptotic Ca(2+) signaling and cell death.

Genome-wide gene expression profiling to predict resistance to anthracyclines in breast cancer patients.

Validated biomarkers predictive of response/resistance to anthracyclines in breast cancer are currently lacking. The neoadjuvant Trial of Principle (TOP) study, in which patients with estrogen receptor (ER)-negative tumors were treated with anthracycline (epirubicin) monotherapy, was specifically designed to evaluate the predictive value of topoisomerase II-alpha (TOP2A) and develop a gene expression signature to identify those patients who do not benefit from anthracyclines. Here we describe in details the contents and quality controls for the gene expression and clinical data associated with the study published by Desmedt and colleagues in the Journal of Clinical Oncology in 2011 (Desmedt et al., 2011). We also provide R code to easily access the data and perform the quality controls and basic analyses relevant to this dataset.

Profile of cancer patients treated at the emergency room of a tertiary cancer care centre in southern Brazil.

The number of cancer care visits in the emergency department for evaluation and treatment has been steadily increasing. Cancer patients represent a challenge for the emergency team because they may have acute symptoms of still undiagnosed malignancy, vague symptoms related to the disease or complications of cancer treatment, all of which can be either mild or potentially life-threatening. The Santo Antonio Hospital is a public institution located in the city of Blumenau (SC, southern Brazil), which serves as a reference centre for cancer care patients for a population of over a million and a half. The objective of this research was to describe the epidemiological profile of the oncologic population who were treated at the emergency department of this hospital, during a period of over six months, starting on April 1st , 2011 to October 31st of the same year. There were 1,051 oncologic visits during this period, with the age ranging from 19 to 89 years. The greater demand for care was sought by patients with urological, breast, upper and lower gastrointestinal tract and lung cancer. The three major complaints of the consultations were pain, respiratory and gastrointestinal symptoms. The visits occurred mainly during daytime mostly on Mondays and Tuesdays. The improvement of primary care services and efficiency of oncology clinics in meeting the high demand for appointments is extremely important, in order to prevent unnecessary visits to the emergency department.

Transcriptional profile of denervated vastus lateralis muscle derived from a patient 8 months after spinal cord injury: a case-report.

A lack of motor neurons abolishes both neurotrophic factor secretion and contractile activity in muscle, which impairs mass, contractile properties, and fibre-type characteristics of the muscle. However, the molecular pathways that can be stimulated or repressed in the scenario of spinal cord injury remain unknown. We investigated for the first time the transcriptional profile of a young male patient 8 months after spinal cord injury. Adaptive metabolic changes of complete denervated skeletal muscle were revealed. In particular, the main molecular pathways involved include metabolic and proteolitic pathways, mitochondrial and synaptic function, calcium homeostasis, sarcomere and anchorage structures. Our data depict the molecular signalling still present in complete denervated skeletal muscle fibres a few months after spinal cord injury. These data could be of interest also to design a specific therapeutic approach aimed at the electrical-stimulation of severe atrophied skeletal muscle.

Effect of phytochemical concentrations on biological activities of cranberry extracts.

Plants of cranberry (Vaccinium macrocarpon) furnish edible fruits and derivates that have been used for the prevention and treatment of urinary tract infections. In the present work we compare two commercial extracts that contain proanthocyanins (PACs) at 4 percent and 20 percent for antimicrobial, antiproliferative, antiradical and protective properties against oxidative stress on cell lines. Both extracts showed antimicrobial activity (MIC values range 3-100 microg/ml). Extract at 20 percent PACs showed higher antiproliferative activity against HepG2 and MCF7 cells, but not against C2C12 cells. Both extracts showed a dose-dependent free-radical scavenging capacity, and a protective effect on the cell damage was also revealed by reduction of intracellular active oxygen species release. Cranberry extracts confirmed antioxidative properties and efficacy in reduction of cell viability that resulted stronger against tumor cells. The pretreatment with cranberry extracts, furthermore, reveal an increase of cell resistance against oxidative stress, suggesting a potential role as a dietary supplement in preventing free-radical damage. The proanthocyanidin content is critical to determine the extract efficacy. In cellular experiments the extracts resulted clearly differentiated in their activity, and the activity was strongly influenced by PACs content. Only in DPPH test the free radical scavenging activity seemed to be directly related to proanthocyanidins content.

Cellular and molecular responses of human skeletal muscle exposed to hypoxic environment.

The effects of a hypobaric, hypoxic environment and exercise performed under extreme conditions, such as at high altitudes, are intriguing physiological aspects that need to be investigated directly on human climbers. Their skeletal muscle is one of the main tissues that can suffer from hypoxia and physical challenges, which will both define the muscle adaptation and the molecular signature of regenerative capacity. We investigated the muscle regenerative capacity characterizing satellite cells. Our study shows that satellite cells are altered by hypobaric, hypoxic environments and exercise performed at high altitudes. Of note, in human skeletal muscle after this 5,000 m a.s.l. expedition, SCs showed a significantly lower ability to regenerate skeletal muscle, in respect to before this high-altitude expedition. This impairment appears to be due to reduced satellite cell activity, consistent with their decreased myogenicity and fusion ability. Furthermore, at the transcriptional level several pathways, such as cell cycle, myogenesis, oxidative metabolism, proteolysis and sarcomeric protein synthesis, were found dysregulated.

Functional assay, expression of growth factors and proteins modulating bone-arrangement in human osteoblasts seeded on an anorganic bovine bone biomaterial.

The basic aspects of bone tissue engineering include chemical composition and geometry of the scaffold design, because it is very important to improve not only cell attachment and growth but especially osteodifferentiation, bone tissue formation, and vascularization. Geistlich Bio-Oss (GBO) is a xenograft consisting of deproteinized, sterilized bovine bone, chemically and physically identical to the mineral phase of human bone. In this study, we investigated the growth behaviour and the ability to form focal adhesions on the substrate, using vinculin, a cytoskeletal protein, as a marker. Moreover, the expression of bone specific proteins and growth factors such as type I collagen, osteopontin, bone sialoprotein, bone morphogenetic protein-2 (BMP-2), BMP-7 and de novo synthesis of osteocalcin in normal human osteoblasts (NHOst) seeded on xenogenic GBO were evaluated. Our observations suggest that after four weeks of culture in differentiation medium, the NHOst showed a high affinity for the three dimensional biomaterial; in fact, cellular proliferation, migration and colonization were clearly evident. The osteogenic differentiation process, as demonstrated by morphological, histochemical, energy dispersive X-ray microanalysis and biochemical analysis was mostly obvious in the NHOst grown on three-dimensional inorganic bovine bone biomaterial. Functional studies displayed a clear and significant response to calcitonin when the cells were differentiated. In addition, the presence of the biomaterial improved the response, suggesting that it could drive the differentiation of these cells towards a more differentiated osteogenic phenotype. These results encourage us to consider GBO an adequate biocompatible three-dimensional biomaterial, indicating its potential use for the development of tissue-engineering techniques.

Evaluation of C-reactive protein and total serum cholesterol in adult patients with bipolar disorder.

The aim of the present study is to evaluate the role of CRP and Total Cholesterol (TC) in patients suffering from type I Bipolar Disorder (BD-I). Moreover, the goal is to elucidate possible CRP and TC differences in different phases of BD-I: acute mania, euthymia and bipolar depression. Medical records of 90 BD-I patients (30 patients with acute mania, 30 in euthymic state, full remission, and 30 in depressive phase) were reviewed to evaluate serum CRP and TC levels. Laboratory data of 30 healthy controls were also obtained. The scores of Young Mania Rating Scale (YMRS), Bech-Rafaelsen Manic Rating Scale (BRMRS) and Hamilton Rating Scale for Depression (HAM-D) were evaluated. CRP levels were higher in acute mania and depressive phase subgroups when compared to healthy controls. CRP was positively associated with BRMRS and YMRS scores in acute mania and with HAM-D in depressive phase subgroups. TC levels were lower in all clinical groups compared to controls. TC levels were negatively correlated to BRMRS, YMRS and HAM-D. In conclusion, the results of the present study support the notion that CRP and TC may be altered in patients with BP-I.

Chemical characteristics and mutagenic activity of PM10 in Torino, a northern Italian city.

Epidemiological studies of particulate matter (PM) have associated PM mass, as well as certain individual components of PM such as secondary particulate with adverse human health effects. For example genotoxic effects attributed to PM may relate to the content of organic compounds but also to the oxidative DNA damage generated by transition metals like iron. However the exact physiochemical mechanism by which PM produces adverse effects is not clear. The aims of this study were to evaluate (1) concentrations of PM10, (2) amounts of bioavailable iron associated with PM10, (3) amounts of secondary particulate expressed as SO4(=) and NO3(-) and (4) the mutagenic activities of PM10 organic extracts. Sampling was carried out in a meteochemical station located in Torino, a northern Italian city with high levels of PM10. The mean PM10 concentration in the considered period was 46.1+/-28.8 microg/m3, the iron mean concentration was 0.83 microg/m3 (+/-0.65 microg/m3) and the bioavailable Fe was 5.7% (+/-4.4%). The data showed that secondary particulate matter (as sum of sulfates and nitrates) constituted about 47% of PM10 total mass. Both iron and secondary species concentrations were positively associated with PM10 levels. Seasonal variations of PM10 concentration, iron level and secondary species amount were significant. Samples were tested for mutagenicity with Salmonella typhimurium strains TA98 and TA100, with and without metabolic activation and a positive response was observed especially for TA98. There were positive statistical associations between mutagenicity and PM10, bioavailable iron, sulfates and nitrates concentrations. Therefore, these results showed the usefulness of this biological approach for monitoring PM10.

Airborne particulate matter: Ionic species role in different Italian sites.

Epidemiological studies have provided evidences for an association between exposure to elevated levels of ambient particulate matter (PM) and increased mortality and morbidity. However, the exact physiochemical nature of the responsible component is not clear. Secondary airborne PM formed from gas-phase pollutants contributes significantly to the most severe particulate air quality events. Although chemical formation for ionic species of aerosol have been observed, they have not been well reported for local variation. This work evaluates the amount of secondary particulate ionic species: sulfates (SO(4)(2-)) and nitrates (NO(3)(-)), chlorides (Cl(-)) and the mutagenic activities of PM10 extracts in different Italian sites (one Southern, one Central and three Northern; in one of the latter also PM2.5 has been evaluated). In general, mean secondary species concentration constitutes about 35-45% of PM10 mass in the North sites, 15% in the center site and 20% in the South site and it is positively associated with PM10 levels. There are significant local differences in the mean levels of PM10 ionic constituents: NO(3)(-) are predominant in northern cities, SO(4)(2-) are more equally distributed and coastal southern city is abundant in Cl(-). Samples were also tested for mutagenicity with Salmonella typhimurium strains TA98 and TA100, with and without metabolic activation; mutagenicity did not correlate with PM10 concentrations. The results showed the important roles and the geographical variability of PM secondary species in the total mass PM10 concentrations and the usefulness of this biological approach for monitoring PM to understand hazards from PM.