ESO-ESMO fifth international consensus guidelines for breast cancer in young women (BCY5).

We dedicate this manuscript in memory of a dear friend and colleague Bella Kaufman. The fifth International Consensus Symposium for Breast Cancer in Young Women (BCY5) took place virtually in October 2020, organized by the European School of Oncology (ESO) and the European Society of Medical Oncology (ESMO). Consensus recommendations for the management of breast cancer in young women were updated from BCY4 with incorporation of new evidence to inform the guidelines. Areas of research priorities as well as specificities in different geographic and minority populations were identified. This manuscript summarizes the ESO-ESMO international consensus recommendations, which are also endorsed by the European Society of Breast Specialists (EUSOMA).

Prevalence of Sexually Transmitted Infections and Factors Associated with HIV Status Among Vulnerable Women in Northern Uganda: Baseline Results from Pe Atye Kena Cohort Study.

BACKGROUND AND OBJECTIVES: HIV infection among vulnerable women (VW) has been attributed to unfavourable power relations and limited access to sexual and reproductive health information and services. This work aims to report sexually-transmitted infections (STI) prevalence and assess the impact of HIV awareness, demographic and socio-behavioural factors on HIV status in a rural area of northern Uganda. METHODS: Pe Atye Kena is a longitudinal cohort intervention study enrolling young women aged 18-49 years old living in the municipality of Gulu, Uganda. HIV, HBV, syphilis serologic tests, and a comprehensive electronic questionnaire on sexual high-risk behaviours were administered before intervention. In this work, we report baseline characteristics of the population along with factors associated with HIV status. Statistical analysis was performed by uni- and multivariable regression models. RESULTS: 461 VW were enrolled (mean age: 29 (SD7.7)). 40 (8.7%) were found to be positive for HIV, 42 (9.1%) for syphilis and 29 (6.3%) for HBV. Older age (> 34 years vs. < 24 years; OR 4.95, 95% CI: 1.7 to 14); having done the last HIV test > 12m before the interview (OR 5.21, 95% CI: 2.3 to 11); suspecting the male sexual partner to be HIV+ (OR 2.2; 95% CI: 1.1 to 4.3); not having used condom at first sexual intercourse (OR 2.6; 95% CI 1.3 to 5.15) were all factors associated with an incident HIV diagnosis. CONCLUSIONS: In this cohort, HIV prevalence is high, and sexual high-risk behaviours are multifaced; future interventions will be aimed to reduce HIV/STIs misconceptions and to promote a sense of community, self-determination and female empowerment.

ESO-ESMO 4th International Consensus Guidelines for Breast Cancer in Young Women (BCY4).

The 4th International Consensus Conference for Breast Cancer in Young Women (BCY4) took place in October 2018, in Lugano, Switzerland, organized by the European School of Oncology (ESO) and the European Society of Medical Oncology (ESMO). Consensus recommendations for the management of breast cancer in young women were updated from BCY3 with incorporation of new evidence to inform the guidelines. Areas of research priorities were also identified. This article summarizes the ESO-ESMO international consensus recommendations, which are also endorsed by the European Society of Breast Specialists (EUSOMA).

Aptamer-based Field-Effect Biosensor for Tenofovir Detection.

During medical treatment it is critical to maintain the circulatory concentration of drugs within their therapeutic range. A novel biosensor is presented in this work to address the lack of a reliable point-of-care drug monitoring system in the market. The biosensor incorporates high selectivity and sensitivity by integrating aptamers as the recognition element and field-effect transistors as the signal transducer. The drug tenofovir was used as a model small molecule. The biointerface of the sensor is a binary self-assembled monolayer of specific thiolated aptamer and 6-mercapto-1-hexanol (MCH), whose ratio was optimized by electrochemical impedance spectroscopy measurements to enhance the sensitivity towards the specific target. Surface plasmon resonance, performed under different buffer conditions, shows optimum specific and little non-specific binding in phosphate buffered saline. The dose-response behavior of the field-effect biosensor presents a linear range between 1 nM and 100 nM of tenofovir and a limit of detection of 1.2 nM. Two non-specific drugs and one non-specific aptamer, tested as stringent control candidates, caused negligible responses. The applications were successfully extended to the detection of the drug in human serum. As demonstrated by impedance measurements, the aptamer-based sensors can be used for real-time drug monitoring.

Surface trap mediated electronic transport in biofunctionalized silicon nanowires.

Silicon nanowires (SiNWs), fabricated via a top-down approach and then functionalized with biological probes, are used for electrically-based sensing of breast tumor markers. The SiNWs, featuring memristive-like behavior in bare conditions, show, in the presence of biomarkers, modified hysteresis and, more importantly, a voltage memory component, namely a voltage gap. The voltage gap is demonstrated to be a novel and powerful parameter of detection thanks to its high-resolution dependence on charges in proximity of the wire. This unique approach of sensing has never been studied and adopted before. Here, we propose a physical model of the surface electronic transport in Schottky barrier SiNW biosensors, aiming at reproducing and understanding the voltage gap based behavior. The implemented model describes well the experimental I-V characteristics of the device. It also links the modification of the voltage gap to the changing concentration of antigens by showing the decrease of this parameter in response to increasing concentrations of the molecules that are detected with femtomolar resolution in real human samples. Both experiments and simulations highlight the predominant role of the dynamic recombination of the nanowire surface states, with the incoming external charges from bio-species, in the appearance and modification of the voltage gap. Finally, thanks to its compactness, and strict correlation with the physics of the nanodevice, this model can be used to describe and predict the I-V characteristics in other nanostructured devices, for different than antibody-based sensing as well as electronic applications.

Nano-fabricated memristive biosensors for biomedical applications with liquid and dried samples.

Nanowire based platforms are widely reported for sensing applications and for their potential in the bio-detection field. In the present work, memristive nanowire devices are implemented for label-free sensing in liquid samples as well as in dried samples via leveraging the modification of the hysteresis in the devices electrical response as a consequence of the surface modification. First, pH sensing in liquid conditions is demonstrated. In addition, the memristive devices are bio-functionalized using antibodies for Prostate Specific Antigen (PSA) as case of study. In this way, the nanowires are converted to memristive biosensors paving the way for future molecular diagnostics applications in general, and for detection of prostate cancer disease in particular.

Study on the bio-functionalization of memristive nanowires for optimum memristive biosensors.

Semiconductor nanowires are emerging as promising building blocks for biosensors enabling direct electrical detection of various biomolecules. In this framework, two-terminal Schottky-barrier silicon (Si) nanowire arrays that exhibit memristive electrical response, so-called memristive devices, are bio-functionalized and converted to memristive biosensors for bio-detection purposes. A comparative analysis of three bio-functionalization strategies is proposed here in order to design and develop optimum memristive biosensors to be implemented in label-free sensing applications. The surface of the device is modified with an anti-free-Prostate Specific Antigen (PSA) antibody as the case of study via: (a) direct adsorption on the device surface, (b) a bio-affinity approach using biotin-streptavidin combination and (c) covalent attachment using (3-glycidyloxypropyl)trimethoxysilane (GPTES). The optimum memristive biosensor is defined via the calibration and comparative study of the biosensors' electrical response under controlled environmental conditions (humidity and temperature) in order to maximize the performance of the biosensor. In addition, it is demonstrated that the direct passive adsorption strategy presents double the performance of the other two methods. The uptake of biological molecules on the nanostructure surface is verified by atomic force microscopy and confocal microscopy. Scanning electron microscopy reveals the details of the surface morphology of the nanofabricated structures before and after bio-functionalization for the three methods applied. The system shows potential for general application in molecular diagnostics, and, in particular, for the early detection of prostate cancer.

Continuous monitoring of Naproxen by a cytochrome P450-based electrochemical sensor.

This paper reports the characterization of an electrochemical biosensor for the continuous monitoring of Naproxen based on cytochrome P450. The electrochemical biosensor is based on the drop-casting of multi-walled carbon-nanotubes (MWCNTs) and microsomal cytochrome P4501A2 (msCYP1A2) on a graphite screen-printed electrode (SPE). The proposed biosensor was employed to monitor Naproxen (NAP), a well-known anti-inflammatory compound, through cyclic voltammetry. The dynamic linear range for the amperometric detection of NAP had an upper limit of 300 µM with a corresponding limit of detection (LOD) of 16 ± 1 µM (S/N=3), which is included in NAP physiological range (9-300 µM). The MWCNT/msCYP1A2-SPE sensor was also calibrated for NAP detection in mouse serum that was previously extracted from mice, showing a slightly higher LOD (33 ± 18 µM). The stability of the msCYP1A2-based biosensor was assessed by longtime continuous cyclic voltammetric measurements. The ability of the sensor to monitor drug delivery was investigated by using a commercial micro-osmotic pump. Results show that the MWCNT/msCYP1A2-SPE sensor is capable of precisely monitoring the real-time delivery of NAP for 16 h. This work proves that the proposed electrochemical sensor might represent an innovative point-of-care solution for the personalization of drug therapies, as well as for pharmacokinetic studies in both animals and humans.

Arthritis induced by adjuvant in spontaneously hypertensive and normotensive rats: endogenous glucocorticoid effects on inflammatory response.

The present study investigated arthritis induced by complete Freund adjuvant (AIA) in spontaneously hypertensive and normotensive rats (respectively, SHR and NTR rats). The inflammatory reaction was studied for 28 days by evaluating paw edema and secondary lesions found 10 days after complete Freund adjuvant (CFA) administration. The body weight of the animals and macroscopic alterations of several organs, including spleen, thymus, adrenal glands, and lymph nodes, were also analyzed. The results showed that the AIA manifestations were decreased in SHRs compared with NTRs. Moreover, this altered inflammatory response was not modified by surgical adrenalectomy.

The looped domain organization of the nucleoid in histone-like protein defective Escherichia coli strains.

We have investigated the major Escherichia coli histone-like proteins (H-NS, HU, FIS, and IHF) as putative factors involved in the maintenance of the overall DNA looped arrangement of the bacterial nucleoid. The long-range architecture of the chromosome has been studied by means of an assay based on in vivo genomic fragmentation mediated by endogenous DNA gyrase in the presence of oxolinic acid. The fragmentation products were analysed by CHEF electrophoresis. The results indicate that in vivo a large fraction of the bacterial chromatin constitutes an adequate substrate for the enzyme. DNA fragments released upon oxo-treatment span a size range from about 1000 kb to a limit-size of about 50 kb. The latter value is in excellent agreement with the average size reported for bacterial chromosomal domains. The DNA gyrase-mediated fragmentation does not appear to be significantly altered in strains depleted in histone-like proteins as compared to an E. coli wild type strain. This suggests that these proteins may not represent critical determinants for the maintenance of the supercoiled loop organisation of the E. coli chromosome.

[Is a totally non-invasive assessment of the hemodynamic profile possible in patients with chronic heart failure?]. / E possibile una valutazione completamente non invasiva del profilo emodinamico dei pazienti con scompenso cardiaco cronico?

Relevant hemodynamic information can be obtained by a comprehensive Doppler echocardiographic examination in patients with various cardiac diseases. The assessment of left heart hemodynamics by Doppler echocardiography has been addressed by several investigators. The feasibility and the accuracy of methods for the estimation of left ventricular filling pressure and cardiac output have been validated by comparative right heart catheterization. Studies have shown that Doppler echocardiography can allow the measurement of pulmonary artery pressures from the pressure gradients across the tricuspid and pulmonary valves. The possibility of completely characterizing cardiac hemodynamics noninvasively has recently been documented: in patients with acute myocardial infarction, automated cardiac output measurement along with the assessment of left ventricular filling by Doppler echocardiography may be used for the identification of hemodynamic subsets. Although Doppler echocardiography can provide noninvasive measures of hemodynamic indices, its value has been disputed since the technique is patient-dependent, time-consuming and requires meticulous acquisition and interpretation by skilled operators. The use of contrast agents may improve the accessibility of both right-sided and left-sided Doppler signals, potentially increasing the number of patients to whom the noninvasive hemodynamic assessment could be applied.

Prognostic value of pulmonary venous flow Doppler signal in left ventricular dysfunction: contribution of the difference in duration of pulmonary venous and mitral flow at atrial contraction.

OBJECTIVES: We assessed the contribution of difference in duration of pulmonary venous and mitral flow at atrial contraction (ARd-Ad) for prognostic stratification of patients with left ventricular (LV) systolic dysfunction. BACKGROUND: Although pulmonary venous flow (PVF) variables may supplement mitral flow patterns in evaluating left ventricular (LV) diastolic function, their value to the prognostic stratification of patients has not been investigated. METHODS: Pulsed wave Doppler mitral and PVF velocity curves were recorded in 145 patients (mean age: 70 years) with LV systolic dysfunction secondary to ischemic or nonischemic cardiomyopathy who were followed for 15 +/- 8 months. In 38% of patients, PVF signal was enhanced by the intravenous (IV) administration of a galactose-based echo-contrast agent. Based on E-wave deceleration time < or = or >130 ms and ARd-Ad, patients were grouped into restrictive (group 1, n = 40), nonrestrictive with ARd-Ad > or =30 ms (group 2, n = 55) and nonrestrictive with ARd-Ad <30 ms (group 3, n = 50). RESULTS: During follow-up, 29 patients died from cardiac causes and 28 were hospitalized for worsening heart failure (HF). On multivariate Cox model, ARd-Ad > or =30 ms provided important prognostic information with regard to cardiac mortality and emerged as the single best predictor of cardiac events (cardiac mortality, hospitalization). The 24-month cardiac event-free survival was best (86.3%) for group 3; it was intermediate (37.9%) for group 2; and it was worst (22.9%) for group 1 (p < 0.0002 group 1 vs. 3; p < 0.0005 group 2 vs. 3; p < 0.0003 group 1 vs. group 2). CONCLUSIONS: Assessment of ARd-Ad exhibited an independent value in the prognostic evaluation of patients with LV systolic dysfunction. Moreover, it contributed to identify patients at low, intermediate and high risk of cardiac events.

Left ventricular size and function in patients with noninsulin-dependent diabetes and postinfarction total or subtotal coronary occlusions.

The aim of this study was to establish the effects of postinfarction total or subtotal coronary occlusion on left ventricular remodeling in patients with noninsulin-dependent diabetes (NIDD) compared with the effects in postinfarct nondiabetic patients. The authors selected 100 patients submitted to coronary angiography between 1 and 5 weeks after acute myocardial infarction (T0: 20.5+/-15.4 days) and classified into three groups: G1: NIDD with coronary occlusion/subocclusion (n=24), G2: controls with coronary occlusion/subocclusion (n=43), G3: controls without coronary occlusion/subocclusion (n=33). At time zero (T0) the following parameters were evaluated: end-systolic and end-diastolic volume indexes (ESVi, EDVi), ejection fraction (EF), echocardiographic wall motion score index (WMI), presence of left ventricular aneurysm, and triple-vessel coronary disease. The frequencies of major cardiovascular events were recorded during follow-up. Significantly greater ESVi and EDVi were noted in G2 compared with G3 (P<0.0001), while no significant differences were observed between NIDD patients and controls. Although left ventricular global and segmental dysfunctions were increased in diabetics, controls with coronary occlusion/subocclusion presented more pronounced EF reduction (P<0.0001 G2 vs G3) and higher elevation in WMI (P<0.005 G2 vs G3). Cardiac events during follow-up were elevated in G1 and G2, particularly as regards the occurrence of congestive heart failure. The authors conclude that NIDD seems to influence in a positive way left ventricular remodeling associated with postinfarct total or subtotal coronary occlusion.

Thermoregulation of Shigella and Escherichia coli EIEC pathogenicity. A temperature-dependent structural transition of DNA modulates accessibility of virF promoter to transcriptional repressor H-NS.

The expression of plasmid-borne virF of Shigella encoding a transcriptional regulator of the AraC family, is required to initiate a cascade of events resulting in activation of several operons encoding invasion functions. H-NS, one of the main nucleoid-associated proteins, controls the temperature-dependent expression of the virulence genes by repressing the in vivo transcription of virF only below a critical temperature (approximately 32 degrees C). This temperature-dependent transcriptional regulation has been reproduced in vitro and the targets of H-NS on the virF promoter were identified as two sites centred around -250 and -1 separated by an intrinsic DNA curvature. H-NS bound cooperatively to these two sites below 32 degrees C, but not at 37 degrees C. DNA supercoiling within the virF promoter region did not influence H-NS binding but was necessary for the H-NS-mediated transcriptional repression. Electrophoretic analysis between 4 and 60 degrees C showed that the virF promoter fragment, comprising the two H-NS sites, undergoes a specific and temperature-dependent conformational transition at approximately 32 degrees C. Our results suggest that this modification of the DNA target may modulate a cooperative interaction between H-NS molecules bound at two distant sites in the virF promoter region and thus represents the physical basis for the H-NS-dependent thermoregulation of virulence gene expression.

A role for H-NS in the regulation of the virF gene of Shigella and enteroinvasive Escherichia coli.

We have investigated the role of H-NS, one of the major components of the bacterial nucleoid, in the expression of the virF gene present on the large virulence plasmid of Shigella and enteroinvasive Escherichia coli in response to different environmental conditions. VirF is an AraC-like protein which activates at least two promoters, virB and virG, both repressed by H-NS. Band shift experiments reveal that the affinity of H-NS for the virF and virB promoters is comparable, while the affinity for the virG promoter is higher. Polyacrylamide gel electrophoresis of three DNA fragments containing the virF, the virB and the VirG promoters demonstrates, in agreement with computer predictions, that they have an intrinsically curved structure, confirming the preference of H-NS for bent DNA. In vivo transcriptional analysis of virF mRNA shows that H-NS negatively controls the expression of virF at 30 degrees C. The expression of a virF-lacZ translational fusion in E.coli wild type and in an hns-defective derivative grown at 30 degrees or 37 degrees C and at pH 6.0 or 7.0 indicates that, in the absence of H-NS, virF expression becomes insensitive to temperature and to limited pH changes. Our results strongly suggest that H-NS controls virF expression by binding to the virF promoter and by repressing its expression at low temperature and at low pH.

[Dipyridamole-echocardiography test in the diagnosis of vasomotor angina]. / Il test ecodipiridamolo nella diagnosi di angina vaso-motoria.

Dipyridamole-atropine echocardiography testing is used extensively for the diagnosis of coronary artery disease and it is highly effective in diagnosing "organic" coronary artery disease by inducing myocardial ischemia via three different mechanisms: maximal coronary artery vasodilatation with phoenomena of flow-maldistribution caused by dipyridamole, increase in myocardial oxygen consumption and reduction of the oxygen supply to the myocardium caused by atropine. Moreover, the abrupt withdrawal of the coronary artery vasodilatation caused by aminophylline, which is routinely infused at the end of the test, may trigger coronary artery spasms in patients with variant angina, thus enhancing the diagnostic power of the test. We report two clinical cases of patients with rest angina and angiographically normal coronary arteries, in whom coronary artery spasm was induced by administering aminophylline during the stress test.

New patterns of family formation in Italy. Which tools for which interpretations?

PIP: This paper presents a detailed discussion of the impact of World War II on subsequent fertility behavior a generation later in Italy. The changes in fertility were delayed and induced by a transgenerational process precipitated by war. War anomie and anomie in the rules of family formation are "symptoms" and not effects of the broad structural changes and historical upheavals during the 30 years of war. Fertility behavior is not construed to be an inevitable outcome of causal processes, but as alternative responses to life situations. The author bases this explanatory model of macrodemographic processes on a variant of Brown and Harris' etiological model that explains the occurrence of depression. It is argued that demographic models of European fertility are needed that acknowledge "a radical form of war that acts on reciprocity systems susceptible to change" and "intervenes to modify the subsequent transgenerational (family) relations." A research design is not now available that relies on the tools of history and anthropology. Thus, the demography of fertility may be reduced to a "mere bookkeeping of vital statistical data or simple economics of resources." The author's new pattern of family formation in Italy considers the "regional family culture and structure to be symptom-formation factors relating to the war and transformations in systems of family relations and exchange." These family changes are linked to war anomie and "changes in the logic of transition strategies from suppression to disconfirmation." War anomie is linked with a generational impact that intervenes in the relationship between changes in the logic of transition strategies and the decline in births. The decline in births is the response to background factors, precipitating events, and symptom-formation or structural factors. The author states that the second demographic transition does not represent a break with prior urban lifestyles and does not modify general trends continuing from the first demographic transition.^ieng

Alternative approaches to fertility transition.

"Using anomalies of the Italian case as the basis, the aim of this article is to verify how the theoretical framework [of demographic transition] put forward by Lesthaeghe can be interpreted.... While the changes in mores in the Italy of the economic boom were a component of the large-scale processes of secularization, rationalization and modernization specific to the [first demographic transition], the practices, values and models of the collective imagination characterizing the second phase of Lesthaeghe's [second demographic transition] are marked by a state of mind that cannot be put down to the 'spirit of the age' during the years of large-scale modernization, but to the emancipatory and 'rational' falling away of many of the barriers to the unfolding of individual life destiny over and above that based on class, gender and age."

Antígenos solúveis liberados por tripomastigotas de Trypanosoma cruzi utilizados no teste de Elisa para detectar cura em pacientes chagásicos após tratamento específico / Soluble antigens released by Trypanosoma cruzi trypomastigotes used Elisa to detect cure in chagasic patients following specific treatment

Dois antígenos solúveis de tripomastigotas do Trypanosoma cruzi, um obtido de sobrenadantes de culturas celulares (AgSb) e o outro excretado/secretado por essas formas em meio de cultura (AgES), foram avaliados em um teste de ELISA para o diagnóstico da infecçao chagásica e controle de cura de pacientes tratados. Os pacientes tratados apresentavam testes de lise mediada pelo complemento e hemoculturas repetidamente negativos, apesar de permanecerem com a sorologia convencional positiva (pacientes dissociados). O teste de lise negativo indica que estes pacientes eliminaram a infecçao. Entre os controles com infecçao ativa, os AgSb e os AgES detectaram respectivamente 93 e 100 por cento dos casos. No entanto, entre os pacientes dissociados, o teste de ELISA, utilizando os AgSb e AgES, foi positivo com 28 por cento e 5 por cento dos soros, respectivamente. Portanto, este teste com os AgES é indicado para o controle de cura da doença de Chagas, podendo vir a substituir a reaçao de lise mediada pelo complemento no acompanhamento sorológico individual de pacientes tratados.

Monoclonal antibodies that recognise filamentous phage: tools for phage display technology.

We generated six hybridoma cell lines that secrete monoclonal antibodies (mAb) which specifically bind filamentous phage coat proteins. Two of these mAb recognise epitopes that include the N terminus of the coat protein III (pIII), while two others are specific for the N terminus of the major coat protein VIII (pVIII). These mAb are valuable tools to study phage assembly and structure. Furthermore, we describe two examples of how these mAb can be exploited in the construction and screening of peptide libraries displayed by the filamentous phase major coat protein. We have used one of these mAb to develop a sensitive ELISA with crude phage supernatants. This assay allows rapid screening of large numbers of clones from random peptide phage libraries. Some of the anti-phage mAb described here can interfere with wild-type phage propagation, while phage carrying modifications in their coat proteins are insensitive to growth inhibition. We have exploited this observation as a tool to favour the growth of phage displaying peptides fused to pVIII, with respect to vector phage.