Combinatorial immune checkpoint blockade increases myocardial expression of NLRP-3 and secretion of H-FABP, NT-Pro-BNP, interleukin-1ß and interleukin-6: biochemical implications in cardio-immuno-oncology.

Background: Immune checkpoint blockade in monotherapy or combinatorial regimens with chemotherapy or radiotherapy have become an integral part of oncology in recent years. Monoclonal antibodies against CTLA-4 or PD-1 or PDL-1 are the most studied ICIs in randomized clinical trials, however, more recently, an anti-LAG3 (Lymphocyte activation gene-3) antibody, Relatlimab, has been approved by FDA in combination with Nivolumab for metastatic melanoma therapy. Moreover, Atezolizumab is actually under study in association with Ipilimumab for therapy of metastatic lung cancer. Myocarditis, vasculitis and endothelitis are rarely observed in these patients on monotherapy, however new combination therapies could expose patients to more adverse cardiovascular events. Methods: Human cardiomyocytes co-cultured with human peripheral blood lymphocytes (hPBMCs) were exposed to monotherapy and combinatorial ICIs (PD-L1 and CTLA-4 or PD-1 and LAG-3 blocking agents, at 100 nM) for 48 h. After treatments, cardiac cell lysis and secretion of biomarkers of cardiotoxicity (H-FABP, troponin-T, BNP, NT-Pro-BNP), NLRP3-inflammasome and Interleukin 1 and 6 were determined through colorimetric and enzymatic assays. Mitochondrial functions were studied in cardiomyocyte cell lysates through quantification of intracellular Ca++, ATP content and NADH:ubiquinone oxidoreductase core subunit S1 (Ndufs1) levels. Histone deacetylases type 4 (HDAC-4) protein levels were also determined in cardiomyocyte cell lysates to study potential epigenetic changes induced by immunotherapy regimens. Results: Both combinations of immune checkpoint inhibitors exert more potent cardiotoxic side effects compared to monotherapies against human cardiac cells co-cultured with human lymphocytes. LDH release from cardiac cells was 43% higher in PD-L1/CTLA-4 blocking agents, and 35.7% higher in PD-1/LAG-3 blocking agents compared to monotherapies. HDAC4 and intracellular Ca++ levels were increased, instead ATP content and Ndufs1 were reduced in myocardial cell lysates (p < 0.001 vs. untreated cells). Troponin-T, BNP, NT-Pro-BNP and H-FABP, were also strongly increased in combination therapy compared to monotherapy regimen. NLRP3 expression, IL-6 and IL-1ß levels were also increased by PDL-1/CTLA-4 and PD-1/LAG-3 combined blocking agents compared to untreated cells and monotherapies. Conclusions: Data of the present study, although in vitro, indicate that combinatorial immune checkpoint blockade, induce a pro- inflammatory phenotype, thus indicating that these therapies should be closely monitored by the multidisciplinary team consisting of oncologists, cardiologists and immunologists.

Sodium-glucose cotransporter 2 inhibitor dapagliflozin prevents ejection fraction reduction, reduces myocardial and renal NF-κB expression and systemic pro-inflammatory biomarkers in models of short-term doxorubicin cardiotoxicity.

Background: Anthracycline-mediated adverse cardiovascular events are among the leading causes of morbidity and mortality in patients with cancer. Sodium-glucose cotransporter 2 inhibitors (SGLT2i) exert multiple cardiometabolic benefits in patients with/without type 2 diabetes, chronic kidney disease, and heart failure with reduced and preserved ejection fraction. We hypothesized that the SGLT2i dapagliflozin administered before and during doxorubicin (DOXO) therapy could prevent cardiac dysfunction and reduce pro-inflammatory pathways in preclinical models. Methods: Cardiomyocytes were exposed to DOXO alone or combined with dapagliflozin (DAPA) at 10 and 100 nM for 24 h; cell viability, iATP, and Ca++ were quantified; lipid peroxidation products (malondialdehyde and 4-hydroxy 2-hexenal), NLRP3, MyD88, and cytokines were also analyzed through selective colorimetric and enzyme-linked immunosorbent assay (ELISA) methods. Female C57Bl/6 mice were treated for 10 days with a saline solution or DOXO (2.17 mg/kg), DAPA (10 mg/kg), or DOXO combined with DAPA. Systemic levels of ferroptosis-related biomarkers, galectin-3, high-sensitivity C-reactive protein (hs-CRP), and pro-inflammatory chemokines (IL-1α, IL-1ß, IL-2, IL-4, IL-6, IL-10, IL-12, IL17-α, IL-18, IFN-γ, TNF-α, G-CSF, and GM-CSF) were quantified. After treatments, immunohistochemical staining of myocardial and renal p65/NF-kB was performed. Results: DAPA exerts cytoprotective, antioxidant, and anti-inflammatory properties in human cardiomyocytes exposed to DOXO by reducing iATP and iCa++ levels, lipid peroxidation, NLRP-3, and MyD88 expression. Pro-inflammatory intracellular cytokines were also reduced. In preclinical models, DAPA prevented the reduction of radial and longitudinal strain and ejection fraction after 10 days of treatment with DOXO. A reduced myocardial expression of NLRP-3 and MyD-88 was seen in the DOXO-DAPA group compared to DOXO mice. Systemic levels of IL-1ß, IL-6, TNF-α, G-CSF, and GM-CSF were significantly reduced after treatment with DAPA. Serum levels of galectine-3 and hs-CRP were strongly enhanced in the DOXO group; on the other hand, their expression was reduced in the DAPA-DOXO group. Troponin-T, B-type natriuretic peptide (BNP), and N-Terminal Pro-BNP (NT-pro-BNP) were strongly reduced in the DOXO-DAPA group, revealing cardioprotective properties of SGLT2i. Mice treated with DOXO and DAPA exhibited reduced myocardial and renal NF-kB expression. Conclusion: The overall picture of the study encourages the use of DAPA in the primary prevention of cardiomyopathies induced by anthracyclines in patients with cancer.

'Candidatus Phytoplasma brasiliense' (16SrXV-A Subgroup) Associated with Cauliflower Displaying Stunt Symptoms in Brazil.

Cauliflower stunt, caused by a phytoplasma of the group 16SrIII-J, was reported in the beginning of 2012 and has occurred with high incidences of infected plants (up to 90%) in crops located in the state of São Paulo in the southeast region of Brazil (3). Diseased plants exhibit general stunting, malformation of inflorescence, reddening leaves, and vessel necrosis (3). Further investigations with plants displaying identical symptoms collected in Nova Bassano, state of Rio Grande do Sul, Brazilian south region, have revealed the presence of a phytoplasma distinct from 16SrIII-J subgroup. Four symptomatic plus four asymptomatic samples were assayed from a field, and the presence of phytoplasma was evidenced by nested PCR assays performed with primers P1/Tint followed by R16F2n/16R2 in three affected plants, which amplified genomic fragments of 1.2 kb from the 16S rRNA gene. No amplification occurred in non-affected samples. Nested PCR products analyzed by conventional RFLP (2) using the enzymes AluI, RsaI, KpnI, HpaII, MseI, HhaI, MboI, and BstUI pointed to the presence of a phytoplasma belonging to group 16SrXV-A in all three phytoplasma-positive samples. Virtual RFLP analysis based on restriction patterns, derived from in silico digestion with 17 endonucleases (4), confirmed the previous results obtained from those samples by conventional RFLP. The 16S rDNA sequences of this phytoplasma identified in cauliflower (GenBank Accession No. JN818845) shared 99% sequence similarity with the reference phytoplasma for subgroup 16SrXV-A (Hibiscus witches'-broom phytoplasma, AF147708), designated 'Candidatus Phytoplasma brasiliense.' Analysis of putative restriction sites showed excellent identity between the phytoplasma studied here and the reference phytoplasma. In addition, the arrangement of branches of a phylogenetic tree constructed with phytoplasmas representing diverse 16Sr groups and subgroups supported that the phytoplasma found in cauliflower is closed related to the representative of the subgroup 16SrXV-A. Association of distinct phytoplasmas with the same kind of disease is not rare and the present pathosystem constitutes a new example. Members of this subgroup have been described almost exclusively in Brazil and previously reported in Sida sp., periwinkle, and hibiscus (1). In some European countries, as well as in the United States and Canada, phytoplasmas belonging to group 16SrI has been associated with this type of disease, which has been reported for various species of the genus Brassica, as published in previous works (3). However, a representative of the group 16SrVI was described in infected plants in Iran (3). Although the 16SrIII-J phytoplasma is currently the most important agent of cauliflower stunt in Brazil, and members of 16SrI are prevalent in other countries, this study revealed that a 16Sr XV-A phytoplasma may be also associated with this important disease of brassicas. Besides, the findings here reported expand the natural host range, including cauliflower as new host for phytoplasmas affiliated with 16SrXV-A. References: (1) B. Eckstein et al. Plant Dis. 95:363, 2009. (2) I. M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (3) M. C. C. Rappussi et al. Eur. J. Plant. Pathol. 133:829, 2012. (4) Wei et al. Int. J. Syst. Evol. Microbiol. 57:1855, 2007.

Interleukin-1 blocking agents as promising strategy for prevention of anticancer drug-induced cardiotoxicities: possible implications in cancer patients with COVID-19.

Cytokines in cardiac tissue plays a key role in progression of cardiometabolic diseases and cardiotoxicity induced by several anticancer drugs. Interleukin-1ß is one on the most studied regulator of cancer progression, survival and resistance to anticancer treatments. Recent findings indicate that interleukin1-ß exacerbates myocardial damages in cancer patients treated with chemotherapies and immune check-point inhibitors. Interleukin1-ß blocking agent canakinumab reduces major adverse cardiovascular events and cardiovascular death in recent cardiovascular trials. We focalized on the main biological functions of interleukin1-ß in cancer and cardiovascular diseases, summarizing the main clinical evidence available to date in literature. Especially in the era of SARS-CoV-2 infection, associated to coagulopathies, myocarditis and heart failure, cancer patients have an increased risk of cardiovascular complications compared to general population, therefore, the pharmacological inhibition of interleukin1-ß should be discussed and considered.

Oncology and Cardiac Rehabilitation: An Underrated Relationship.

Cancer and cardiovascular diseases are globally the leading causes of mortality and morbidity. These conditions are closely related, beyond that of sharing many risk factors. The term bidirectional relationship indicates that cardiovascular diseases increase the likelihood of getting cancer and vice versa. The biological and biochemical pathways underlying this close relationship will be analyzed. In this new overlapping scenario, physical activity and exercise are proven protective behaviors against both cardiovascular diseases and cancer. Many observational studies link an increase in physical activity to a reduction in either the development or progression of cancer, as well as to a reduction in risk in cardiovascular diseases, a non-negligible cause of death for long-term cancer survivors. Exercise is an effective tool for improving cardio-respiratory fitness, quality of life, psychological wellbeing, reducing fatigue, anxiety and depression. Finally, it can counteract the toxic effects of cancer therapy. The protection obtained from physical activity and exercise will be discussed in the various stages of the cancer continuum, from diagnosis, to adjuvant therapy, and from the metastatic phase to long-term effects. Particular attention will be paid to the shelter against chemotherapy, radiotherapy, cardiovascular risk factors or new onset cardiovascular diseases. Cardio-Oncology Rehabilitation is an exercise-based multi-component intervention, starting from the model of Cardiac Rehabilitation, with few modifications, to improve care and the prognosis of a patient's cancer. The network of professionals dedicated to Cardiac Rehabilitation is a ready-to-use resource, for implementing Cardio-Oncology Rehabilitation.

[Body profile and physical and cognitive function by age in ambulatory elderly women from the city of Córdoba]. / Perfil corporal y función física y cognitiva según edad en mujeres mayores ambulatorias de la ciudad de Córdoba.

UNLABELLED: Aging produces body changes such as redistribution of fat and loss of muscle mass and strength, predisposing to fragility, functional impairment and disability. OBJECTIVE: To analyze the relationship between body profile and physical and cognitive function by age in in ambulatory elderly women from the city of Córdoba. METHODS: 178 healthy older women (OW) ≥60 years free living were evaluated attending centers of retirees and day homes in the city of Córdoba. We evaluated body profile from: skeletal muscle mass index (SMMI), relative body adiposity (RBA) -dual X-ray absorptiometry- and muscle strength (MS) -dynamometry-. Categories: normal/(N) sarcopenia/(SP), obesity/(OB), sarcopenic obesity/(SO); Physical function: with/without physical limitation (PL); cognitive function: with/without cognitive impairment (CI). INSTRUMENTS: Lunar Prodigy Densitometer and Smedley dynamometer, Lawton and Brody and Minimental Examination of Folstein scales. RESULTS: SO prevailed and increased with age, contrary to OB. Most of the OW did not PL or CI. Only 2.25% had low SMMI and 48.3% dynapenia. 76.97% had elevated RBA. The SP - obese or not - had greater PL and CI. CI frequency doubled to PL (15.17% versus 6.74%). We found negative correlations and significant associations between age and MS (r= -0.279; p=0.0001), physical function (r=-0.164; p=0.0283) and cognitive function (r=-0.028; p=0.0002). CONCLUSIONS: In this group of healthy OW the dynapenia was responsible for the observed SP, not low SMMI. The OW with SP had more PL and CI, and increased with age.

El envejecimiento produce cambios corporales como redistribución de la grasa y pérdida de la masa y fuerza muscular, predisponiendo a fragilidad, deterioro funcional y discapacidad. Objetivo: Analizar la relación entre perfil corporal y función física y cognitiva según edad en mujeres mayores de la ciudad de Córdoba. Material y Métodos: Se valoraron 178 Mujeres Mayores (MM) sanas ambulatorias ≥60 años que asisten a Centros de Jubilados y Hogares de Día de Córdoba capital. Se evaluó Perfil Corporal a partir de: índice de masa muscular esquelética (IMME), adiposidad corporal relativa (ACR) -absorciometría dual de rayos X- y fuerza muscular (FM) ­dinamometría-. Categorías: Normal/(N) Sarcopenia/(SP), Obesidad/(OB), Obesidad Sarcopénica/(OB/SP); Función Física: con/sin limitación física (LF); y Función Cognitiva: con/sin deterioro cognitivo (DC). Instrumentos: Densitómetro Lunar Prodigy y dinamómetro Smedley, Escalas de Lawton y Brody y Minimental Examination de Folstein. Resultados: La OB/SP predominó y se incrementó con la edad; contrario a la OB. La mayoría de las MM no presentó LF ni DC. Sólo 2,25% tuvo IMME bajo y 48,3% dinapemia. El 76,97% tenía ACR elevada. Las SP ­obesas o no- presentaron mayor LF y DC. La frecuencia de DC duplicó a la de LF (15,17% versus 6,74%). Se hallaron correlaciones negativas y asociaciones significativas entre edad y FM (r= -0,279; p=0,0001), FF (r= -0,164; p=0,0283) y FC (r= -0,028; p=0,0002). Conclusiones: En este grupo de MM sanas la dinapenia fue responsable de la SP observada, no el IMME bajo. Las ancianas SP tuvieron mayor LF y DC, y aumentaron con la edad.

Diagnostic specificity of serum chromogranin-A for pheochromocytoma in patients with renal dysfunction.

We assessed the clinical utility of serum chromogranin A (CgA) alone and in combination with plasma catecholamines in the diagnosis of pheochromocytoma in patients with mild to moderate renal impairment. The study population consisted of 44 normal subjects, 50 subjects with proven pheochromocytoma, and 82 subjects with hypertension (60 primary and 22 secondary) suspected but not proven to have the disease. In this highly selected group with high disease prevalence (38%), the overall sensitivity, specificity, accuracy, and positive and negative predictive values of serum CgA in the diagnosis of pheochromocytoma were 86%, 74%, 79%, 67%, and 94%, respectively. However, in patients with creatinine clearance less than 1.33 mL/s (80 mL/min), these values dropped to 85%, 50%, 59%, 38%, and 90%, respectively. The combination of plasma catecholamines and CgA provided the best overall specificity (95%), accuracy (88%), and positive predictive value (91%). In patients with a creatinine clearance of more than 1.33 mL/s, the combination gave a specificity of 98%, an accuracy of 89%, and a positive predictive value of 97%. These results show that serum CgA has poor diagnostic specificity in the diagnosis of pheochromocytoma when renal function is impaired. Combining CgA with plasma catecholamines provides a lower sensitivity, but excellent specificity, accuracy, and positive predictive value.

HLA-DQA1 allele and genotype frequencies in a northern Italian population.

HLA-DQA1 typing of 227 randomly selected Northern Italian people by the use of polymerase chain reaction are reported. The combined use of commercial Amplitype HLA-DQalpha system and four sequence-specific oligonucleotide probes allows the definition of 8 alleles and 36 genotypes, arranged according to World Health Organisation nomenclature. Seven of these genotypes are not observed among the analyzed samples. Allele frequencies range from 1.5 to 35.7% and genotype observations do not deviate significantly from Hardy-Weinberg equilibrium; observed heterozygosity is 0.8238 with an allelic diversity value of 0.79 and the power of discrimination is 0.925. Our Italian population sample shows differences from other Caucasian samples both for allele and genotype frequencies. This locus typing for the 8 defined alleles provides a rapid and sensitive method in individual identification and paternity investigation.

HLA-DQA1 and amelogenin coamplification: a handy tool for identification.

A protocol for HLA-DQA1 and gender identification by single amplification is described. The use of the commercial HLA-DQA1 amplification kit (Perkin Elmer) permits a positive response for sex determination by adding primers for a short sequence on the first intron of Amelogenin gene. The suggested amplification protocol results in PCR products easily and clearly detectable on ethidium bromide stained agarose gel or silver stained polyacrylamide gel. In both gels the HLA-DQA1 observations at 242-239 bp are accomplished with a single band at 106 bp in females and a doublet 112-106 bp in males. HLA-DQA1 reverse dot-blot hybridization is unaffected by the presence of X and Y amplified fragments.

[Carotid endarterectomy by eversion and reimplantation. Techniques and results]. / L'endartérectomie carotidienne par éversion et réimplantation. Techniques et résultats.

Endarterectomy of the carotid bifurcation through a longitudinal arteriotomy of the common and internal carotid arteries followed by a direct closure is still accepted as the main surgical technique for carotid stenoses. However, this technique is complicated in about 10% of the cases by a restenosis. Mechanisms of myointimal hyperplasia leading to restenosis are not completely explained. The technique of eversion endarterectomy of the internal carotid artery has been said to be an alternate technique which could decrease the incidence of restenosis. We described the three main techniques of eversion endarterectomy and their results. The technique of eversion endarterectomy after proximal section of the internal carotid artery is the most frequently used. It allows to treat easily length excess of the internal carotid artery. Its results are good in terms of neurologic morbidity and mortality. However, randomized comparative studies are still required in order to determine its role on the incidence of restenosis despite the first encouraging results of first non comparative studies. The technique of eversion endarterectomy after distal section of the internal carotid artery has been proposed by Chevalier who reported also good short term results and no delayed restenosis.

[Extra-popliteal localizations of adventitial cysts. Review of the literature]. / Les localisations extra-poplitées des kystes adventitiels. Revue de la littérature.

Cystic adventitial disease is an uncommon vascular anomaly first described in 1947 in the external iliac artery. The preferential localization is the popliteal artery although other arteries and veins may also be involved though not always recognized. There have been 45 extra-popliteal localizations of adventitial cysts reported in the literature. Thirty cases involved an artery an 15 a vein. The iliofemoral axis, with 33 reported cases, is the preferential localization of these extra-popliteal adventitial cysts (including 22 arteries and 11 veins). All the other cases also involved a vessel near an articulation (knee, ankle, elbow, wrist). Despite a preferential popliteal arterial localization only one case involved the popliteal vein. The diagnosis is rarely made before surgery, probably because of the nonspecific clinical presentation. Ultrasonography should allow better recognition of these adventitial cysts, eliminating an aneurysm or a synovial cyst, and evidencing the localization of the cyst within the vessel wall.

[Primary carcinoma of the gallbladder: ultrasonic features (remarks on 8 cases)]. / Il carcinoma primitivo della colecisti: aspetti ecotomografici (considerazioni su otto casi).

The authors illustrate the ultrasound features of gallbladder carcinoma after reviewing literature and basing upon their own experience. The diagnosis of this neoplasia is formulated more frequently today by introduction of scanning means, but it remains very difficult on initial phase because of its precocious lymphatic and hilar infiltration.

[Epidemiology and monitoring of closed abdominal injuries]. / Epidemiologia e monitoraggio dei traumi addominali chiusi.

The authors review the epidemiological and aetiopathogenetic characteristics of abdominal blunt injuries (118 cases) monitored in a homogeneous manner. The relatively young age of the patients, the diagnostic difficulties encountered and the frequent presence of associated lesions call for the rigorous application of a predetermined clinico-diagnostic protocol.

[The treatment of bacterial infections of the lower respiratory tract in childhood. An open comparative study of sulbactam/ampicillin vs. ceftazidime]. / Trattamento delle infezioni batteriche delle basse vie respiratorie in età pediatrica. Studio aperto comparativo sulbactam/ampicillin vs ceftazidime.

Sixty children suffering from lower respiratory tract infections have been included into this study, respectively 30 (18 M + 12 F) in the sulbactam/ampicillin (S/A) group and 30 (20 M + 10 F) in the ceftazidime (CFT) group. Average age was 42.9 months +/- 34.4 in S/A group (range 6-120) and 48.7 +/- 42.1 (range 6-144) in CFT group. Both groups were similar as to sex, age, type and duration of the infection. Posology was 150 mg/kg/die for S/A and 50 mg/kg/die for CFT. The duration of treatment was 7.2 days +/- 2.2 (range 5-12) for S/A group and 6.4 days +/- 1.6 (range 5-12) for CFT group. At the end of the therapy clinical recovery has been obtained in all cases. A rapid defervescence and remission of symptoms at an identical rate has been recorded in both treatment groups. General and local tolerability was excellent in both treatment groups.

[Taste disorders in forensic medicine]. / Alterazioni del gusto in medicina legale.

Taste disorders can a rise from lesions of peripheral receptors, taste control pathways or cortical area involvement. Among peripheral lesions, trauma of the tongue and oropharynx are the most common. Iatrogenic lesions of facial and glossopharingeal nerves are very important in Forensic Medicine, while there are different opinions about taste alterations due to head injuries; hypogeusia associated to smell disorders are found in 0.4-0.5% of patient after head trauma with good prognosis (90% healing) while qualitative disorders are more common (30%). The Authors describe clinical methodologies for taste evaluation and their application in Forensic Medicine. Forensic estimation of taste disorders con be classified by two main groups: study of cause relation between the occurrence and damage and quantitative valuation of the damage in three different juridical ambits: Penal, Civil, Insurance and foresight. In Penal Right taste damages could be classified among personal lesion crimes and can be classified as serious (permanent injury of taste) ore very serious (complete lost of taste function). Italian Legislation equipare the 5 sense organs. In Civil Right evaluation the so-called "biologic damage" and working ability are considered; this means very different evaluations. In the most recent baremes, generic damage is estimated by different Authors from 0 to 10% while with regard to specific working capacity, common evaluation criteria does not exist. In Insurance taste disorders evaluation is based only on working ability and not on biologic damage. In the previdenzial ambit, taste disorders are not even included in the most recent tables of permanent invalidity estimation. The Authors propose new and more efficacious valutation criteria for taste disorders in all ambits, hoping for more interest in the Forensic aspects of taste, a too often forgotten sensory function.

[Endoscopic polypectomy and adenoma-carcinoma sequence of the large intestine. Considerations on personal case series]. / Polipectomia endoscopica e sequenza adenoma-carcinoma nel grosso intestino. Considerazioni su casistica originale.

Since the introduction of colonoscopic polypectomy, the management of colonic polyps has dramatically changed. What once was a major transabdominal operation now is a routine therapeutic endoscopic procedure. To test the hypothesis of the adenoma-carcinoma sequence histopathologic features of polypoid lesions are retrospectively evaluated. In our experience with 566 polypectomies 430 adenomatous polyps, 71 hyperplastic, 58 inflammatory polyps, and 4 juvenile lesions are reported. Only adenomatous polyps showed dysplasia. Overall, severe and moderate dysplasia was respectively observed in 6.7% and 23% of the adenomas. Infiltrating carcinoma was present in 2.2% of the cases. Villous adenomas showed high rates of severe dysplasia (37%) and invasive carcinoma (18%). Location and sex did not seem to have a role in the malignant potential of adenomas.

Aluminium, carbonyls and cytokines in human nipple aspirate fluids: Possible relationship between inflammation, oxidative stress and breast cancer microenvironment.

The human breast is likely exposed to Al (aluminium) from many sources including diet and personal care products. Underarm applications of aluminium salt-based antiperspirant provide a possible long-term source of exposure, especially after underarm applications to shaved and abraded skin. Al research in breast fluids likely reflects the intraductal microenvironment. We found increased levels of aluminium in noninvasively collected nipple aspirate fluids (NAF) from 19 breast cancer patients compared with 16 healthy control subjects (268 vs 131 µg/l, respectively; p < 0.0001). In the same NAF samples we found significantly increased levels of protein oxidative carbonyls in cancer patients compared to healthy women (2.35 vs 0.41 nmol/mg prot, respectively; p < 0.0001). Aluminium content and carbonyl levels showed a significant positive linear correlation (r(2) 0.6628, p < 0.0001). In cancer NAF samples (containing higher amounts of aluminium salts) we also found a significantly increased levels of pro-inflammatory cytokines (IL-1ß, IL-6, IL-12 p70, and TNF-α) and chemoattractant CC and CXC chemokines (IL-8, MIP-1α and MCP-1). In 12 invasive cancer NAF samples we found a significant positive linear correlation among aluminium, carbonyls and pro-inflammatory IL-6 cytokine (Y = 64.79x-39.63, r(2) 0.8192, p < 0.0005), as well as pro-inflammatory monocyte chemoattractant MCP-1 cytokine (Y = 2026x-866, r(2) 0.9495, p < 0.0001). In addition to emerging evidence, our results support the possible involvement of aluminium ions in oxidative and inflammatory status perturbations of breast cancer microenvironment, suggesting aluminium accumulation in breast microenvironment as a possible risk factor for oxidative/inflammatory phenotype of breast cells.

Chromatography by silica-gel chromatobars. Prospects of its application in the field of forensic toxicology.

A new type of chromatographic column of silica gel 20 mm in diameter and 180 mm long has been investigated in the field of forensic analytical toxicology. In order to evaluate the usefulness of chromatobars in isolating organic compounds from biological extracts, sodium phenyl-ethyl-barbiturate, atropine, morphine and strychnine were added to the aqueous extract obtained with Stas-Otto's isolation method. Afterwards the residues of the acidic ether extract, alkaline ether extract and alkaline chloroform extract, were chromatographed on chromatobar. This chromatographic device seems able to avoid the drawbacks and limitations of paper and thin-layer chromatography, mainly from the preparative aspect. The method is particularly advantageous in analysing the whole extract obtained with the Stas-Otto process, without bringing about its partition, especially when slight traces of toxic substances are present.