AI-aided holographic flow cytometry for label-free identification of ovarian cancer cells in the presence of unbalanced datasets.

Liquid biopsy is a valuable emerging alternative to tissue biopsy with great potential in the noninvasive early diagnostics of cancer. Liquid biopsy based on single cell analysis can be a powerful approach to identify circulating tumor cells (CTCs) in the bloodstream and could provide new opportunities to be implemented in routine screening programs. Since CTCs are very rare, the accurate classification based on high-throughput and highly informative microscopy methods should minimize the false negative rates. Here, we show that holographic flow cytometry is a valuable instrument to obtain quantitative phase-contrast maps as input data for artificial intelligence (AI)-based classifiers. We tackle the problem of discriminating between A2780 ovarian cancer cells and THP1 monocyte cells based on the phase-contrast images obtained in flow cytometry mode. We compare conventional machine learning analysis and deep learning architectures in the non-ideal case of having a dataset with unbalanced populations for the AI training step. The results show the capacity of AI-aided holographic flow cytometry to discriminate between the two cell lines and highlight the important role played by the phase-contrast signature of the cells to guarantee accurate classification.

A multicentre, open-label, single-arm phase II trial of the efficacy and safety of sclerotherapy using 3% polidocanol foam to treat second-degree haemorrhoids (SCLEROFOAM).

BACKGROUND: The aim of the present study was to evaluate the efficacy and safety of 3% polidocanol foam for treating 2nd-degree haemorrhoids. METHODS: A multicentre, open-label, single-arm, phase 2 trial involving 10 tertiary referral centres for haemorrhodal disease (HD) was performed. Between January and June 2019, patients with 2nd-degree haemorrhoids were prospectively included in this study. The primary outcome was to establish the success rate after one sclerotherapy session in terms of complete resolution of bleeding episodes one week after the injection. The Hemorrhoidal Disease Symptom Score (HDSS), the Short Health Scale for HD (SHS-HD) score and the Vaizey incontinence score were used to assess symptoms and their impact on quality of life and continence. Pain after the procedure, subjective symptoms and the amount and type of painkillers used were recorded. Patients were followed up for 1 year. RESULTS: There were 183 patients [111 males; 60.7%, mean age 51.3 ± 13.5 (18-75) years]. Complete resolution of bleeding was reached in 125/183 patients (68.3%) at 1 week and the recurrence rate was 12% (15/125). Thirteen patients (7.4%) underwent a second sclerotherapy session, while only 1 patient (1.8%) had to undergo a third session. The overall 1-year success rate was 95.6% (175/183). The HDSS and the SHS score significantly improved from a median preoperative value of 11 and 18 to 0 and 0, respectively (p < 0.001). There were 3 episodes of external thrombosis. No serious adverse events occurred. CONCLUSIONS: Sclerotherapy with 3% polidocanol foam is a safe, effective, painless, repeatable and low-cost procedure in patients with bleeding haemorrhoids.

Phase diagram of the NaCl-water system from computer simulations.

NaCl aqueous solutions are ubiquitous. They can crystallize into ice, NaCl, or NaCl · 2H2O depending on the temperature-concentration conditions. These crystallization transitions have important implications in geology, cryopreservation, or atmospheric science. Computer simulations can help understand the crystallization of these solids, which requires a detailed knowledge of the equilibrium phase diagram. We use molecular simulations in which we put at contact the solution with the solid of interest to determine points of the solid-solution coexistence lines. We follow two different approaches, one in which we narrow down the melting temperature for a given concentration and the other in which we equilibrate the concentration for a given temperature, obtaining consistent results. The phase diagram thus calculated for the selected model (TIP4P/2005 for water molecules and Joung-Cheatham for the ions) correctly predicts coexistence between the solution and ice. We were only able to determine NaCl · 2H2O-solution coexistence points at higher temperatures and concentrations than in the experiment, so we could not establish a direct comparison in this case. On the other hand, the model underestimates the concentration of the solution in equilibrium with the NaCl solid. Our results, alongside other literature evidence, seem to indicate that ion-ion interactions are too strong in the model. Our work is a good starting point for the improvement of the potential model and for the study of the nucleation kinetics of the solid phases involved in the phase diagram.

Effect of task complexity on motor and cognitive preparatory brain activities.

In the present study, we investigated scalp-recorded activities of motor and cognitive preparation preceding stimulus presentation in relatively simple and complex visual motor discriminative response tasks (DRTs). Targets and non-targets were presented (with equal probability) in both tasks, and the complexity of the task depended on the discrimination and categorization processing load, which was based on the number of stimuli used (two stimuli in the simple- and four in the complex-DRT, respectively). We recorded event-related potentials (ERPs) in 16 participants in simple-DRT and 16 participants in complex-DRT. At the behavioral level, the performance was faster and more accurate in simple-DRT. Two pre-stimulus ERPs were considered: the central Bereitschaftspotential (BP) and the prefrontal negativity (pN). Both components showed earlier onset and larger amplitude in the complex-DRT. Overall, the simple-DRT required less motor and cognitive preparation in premotor and prefrontal areas compared to the complex-DRT. Present findings also suggest that the pN component was not reported in previous studies, likely because most ERP literature focusing on pre-stimulus ERP used simple-DRTs, and with such a task the pN amplitude is small and can easily go undetected.

Females are more proactive, males are more reactive: neural basis of the gender-related speed/accuracy trade-off in visuo-motor tasks.

In the present study, we investigated neural correlates associated with gender differences in a simple response task (SRT) and in a discriminative response task (DRT) by means of event-related potential (ERP) technique. 120 adults participated in the study, and, based on their sex, were divided into two groups matched for age and education level. Behavioral performance was assessed with computing response speed, accuracy rates and response consistency. Pre- and post-stimulus ERPs were analyzed and compared between groups. Results indicated that males were faster than females in all tasks, while females were more accurate and consistent than males in the more complex tasks. This different behavioral performance was associated with distinctive ERP features. In the preparation phase, males showed smaller prefrontal negativity (pN) and visual negativity (vN), interpreted as reduced cognitive preparation to stimulus occurrence and reduced reliance on sensory proactive readiness, respectively. In the post-stimulus phase, gender differences were present over occipital (P1, N1, P2 components) and prefrontal (pN1, pP1, pP2 components) areas, suggesting allocation of attentional resources at distinct stages of information processing in the two groups. Overall, the present data provide evidence in favor of a more proactive and cautious cognitive processing in females and a more reactive and fast cognitive processing in males. In addition, we confirm that (1) gender is an important variable to be considered in ERP studies on perceptual processing and decision making, and (2) the pre-stimulus component analysis can provide useful information concerning neural correlates of upcoming performance.

Modality-specific sensory readiness for upcoming events revealed by slow cortical potentials.

Human brain activity allows to anticipate future events and to prepare the next action accordingly; consistently, event-related potential (ERP) studies found action preparatory brain activities in the premotor and prefrontal cortex. In the present study, we investigated the preparatory activity in the sensory cortical regions. Slow cortical potentials were recorded during passive tasks, i.e., subjects expected for a sensory stimulus and no motor or cognitive response were required. In particular, we tested the hypothesis that perceptual anticipatory cortical mechanisms were modality specific. Three groups of 21 young adults underwent passive perceptual tasks in different sensory modalities (visual, auditory, or somatosensory). We confirmed the presence of a visual negativity (vN) component for the visual modality starting about 800 ms before stimulus with source in extrastriate areas and we found novel modality-specific sensory readiness components for the auditory and somatosensory modalities. The auditory positivity (aP) started about 800 ms before stimulus with source in bilateral auditory cortices and the somatosensory negativity (sN) started about 500 ms before stimulus with source in the somatosensory secondary cortex, contralateral to the stimulated hand. The scalp topography and intracranial sources of these three slow preparatory activities were mirrored with inverted polarity at early post-stimulus stage evoking the well-known visual P1, auditory N1, and somatosensory P100 components. Present findings contribute to widening the family of slow wave preparatory components, providing evidence about the relationship between top-down and bottom-up processing in sensory perception.

Weak proactive cognitive/motor brain control accounts for poor children's behavioral performance in speeded discrimination tasks.

BACKGROUND: Motor and inhibitory control rely on frontal cortex activity, which is known to reach full maturation only in late adolescence. The development of inhibitory control has been studied using event-related potentials (ERP), focusing on reactive processing (i.e. the N2 and the P3 components). Scarce information exists concerning pre-stimulus activity as that represented by the Bereinshafstpotential (BP) and by the prefrontal negativity (pN). Further, no literature exists concerning the post-stimulus components originating within the anterior insula (pN1, pP1, pP2). This study aims at associating children performance with these motor-cognitive processing in frontal brain areas. METHODS: High-resolution EEG recordings were employed to measure ERPs from 18 children (12 years old) and 18 adults (28 years old) during a visuo-motor discriminative response task. Response time (RT), commission (CE) and omission errors, and RT variability were compared between groups. At brain level, two pre-stimulus (BP and pN) and seven post-stimulus (P1; pN1; N1; pP1; N2; pP2; P3) ERP components were compared between groups. RESULTS: Children showed slower and more variable RTs and poorer inhibition (higher CEs) than adults. At electrophysiological level, children presented smaller BP and pN. After stimulus onset, children showed lower amplitude of N1, pP1, P3, and pP2 components. The P1, pP1, N2 and P3 were delayed compared to adults. CONCLUSIONS: Our results demonstrate that children are characterized by less intense task-related proactive activities in frontal cortex, which may account for subsequent poor and delayed reactive processing and, thus, for inaccurate and slow performance.

Resolution gain in space-time digital holography by self-assembling of the object frequencies.

Space-time digital holography (STDH) exploits the object motion to record the hologram in a hybrid ST domain. This representation adds new capabilities to conventional DH, e.g., unlimited field of view and variable phase shifting. This is the best candidate for imaging biological samples flowing in microfluidic channels. Here, we show that STDH is able to improve the spatial resolution as well. Different from other super-resolution approaches, stitching between holograms or their spectra is no longer required. Moreover, we introduce a new oblique STDH modality to record and process hybrid ST representations. This allows improving resolution in 2D with one single object scan, paving the way to the use of STDH for super-resolution imaging onboard microfluidic devices.

Exercise-related cognitive effects on sensory-motor control in athletes and drummers compared to non-athletes and other musicians.

Both playing a musical instrument and playing sport produce brain adaptations that might affect sensory-motor functions. While the benefits of sport practice have traditionally been attributed to aerobic fitness, it is still unknown whether playing an instrument might induce similar brain adaptations, or if a specific musical instrument like drums might be associated to specific benefits because of its high energy expenditure. Since the aerobic costs of playing drums was estimated to be comparable to those of average sport activities, we hypothesized that these two groups might show both behavioral and neurocognitive similarities. To test this hypothesis, we recruited 48 young adults and divided them into four age-matched groups: 12 drummers, 12 athletes, 12 no-drummer musicians and 12 non-athletes. Participants performed a visuo-motor discriminative response task, namely the Go/No-go, and their cortical activity was recorded by means of a 64-channel electroencephalography (EEG). Behavioral performance showed that athletes and drummers were faster than the other groups. Electrophysiological results showed that the pre-stimulus motor preparation (i.e. the Bereitschaftspotential or BP) and attentional control (i.e., the prefrontal negativity or pN), and specific post-stimulus components like the P3 and the pP2 (reflecting the stimulus categorization process) were enhanced in the athletes and drummers' groups. Overall, these results suggest that playing sport and drums led to similar benefits at behavioral and cognitive level as detectable in a cognitive task. Explanations of these findings, such as on the difference between drummers and other musicians, are provided in terms of long-term neural adaptation mechanisms and increased visuo-spatial abilities.

The proactive self-control of actions: Time-course of underlying brain activities.

Proactive brain control optimizes upcoming actions and inhibits unwanted responses. In the present event-related potential (ERP) study, participants freely decided in advance whether to respond or not to an upcoming stimulus, then prepared or not the action according to their decision; finally, a stimulus was delivered, and subjects had to respond (or not). During the decision-making stage, a prefrontal negativity raised bilaterally in case no-response was decided, reflecting the first brain signal of proactive inhibition. Simultaneously, slow activity raised over premotor cortices independently from the decision taken, and then raised during the preparation phase only in the case of response decision (as a sort of accelerator). When the decision was not to respond, the prefrontal activity remained sustained (as a sort of brake) and showed a right-lateralized distribution during the preparation phase. Overall, we described the time-course of a proactive accelerating-braking system regulating self-control of actions.

Meningococcal carriage in children and young adults in the Philippines: a single group, cross-sectional study.

This cross-sectional prevalence study investigates meningococcal carriage for the first time in a Southeast Asian population. Posterior pharyngeal swabs were collected between August 2013 and March 2014 from 937 healthy Filipinos aged 5-24 years attending school or university in Manila. Of these, 35 were found to be carriers giving an overall carriage prevalence of 3·7% [95% confidence interval (CI) 2·6-5·2]. Carriage was associated with age (P < 0·001) and was highest (9·0%, 95% CI 5·5-13·8) in subjects aged 10-14 years, but was comparatively low (<3%) in all other age groups considered. This suggests that an immunization programme in the Philippines designed to reduce carriage acquisition and induce herd immunity may require a vaccine dose before the age of 10 years. Serogroup B was most commonly carried (65·7%, 95% CI 47·8-80·9), with a small number of carriers for serogroups C, Y and W also present. Two individuals (5·7%, 95% CI 0·7-19·2) who were simultaneously carrying multiple serogroups were identified. This exploratory study provides valuable insight into the asymptomatic carriage of Neisseria meningitidis in a healthy subset of the Filipino population and illustrates the importance of generating local carriage data.

Forward electrohydrodynamic inkjet printing of optical microlenses on microfluidic devices.

We report a novel method for direct printing of viscous polymers based on a pyro-electrohydrodynamic repulsion system capable of overcoming limitations on the material type, geometry and thickness of the receiving substrate. In fact, the results demonstrate that high viscosity polymers can be easily manipulated for optical functionalizing of lab-on-a-chip devices through demonstration of direct printing of polymer microlenses onto microfluidic chips and optical fibre terminations. The present system has great potential for applications from biomolecules to nano-electronics. Moreover, in order to prove the effectiveness of the system, the optical performance of such microlenses has been characterized by testing their imaging capabilities when the fibroblast cells were allowed to flow inside the microfluidic channel, showing one of their possible applications on-board a LoC platform.

Self-propelling bacteria mimic coherent light decorrelation.

We show here that live e-coli bacterial culture, thanks to the self-propelling feature, can significantly reduce the coherent noise. In fact, the typical self-propelled drive of such microorganisms provides enough time diversity in speckle patterns. Optical properties of a bacteria suspension have been investigated and analyzed thus showing that it behaves as a quite good optical speckle decorrelation device. Samples with different bacteria densities have been studied. The decorrelation effect has been demonstrated by probing the imaging performance in through transmission in coherent microscope configuration.

Optofluidic holographic microscopy with custom field of view (FoV) using a linear array detector.

Simple and effective imaging strategies are of utmost interest for applications on a lab-on-chip scale. In fact, the majority of diagnostic tools for medical as well as biotechnological studies still employ image-based approaches. Having onboard the chip a compact but powerful imaging apparatus with multiple imaging capabilities, such as 3D dynamic focusing along the optical axis, unlimited field of view (FoV) and double outputs, namely, intensity and quantitative phase-contrast maps of biological objects, is of extreme importance for the next generation of Lab-on-a-Chip (LoC) devices. Here we present a coherent 3D microscopy approach with a holographic modality that is specifically suitable for studying biological samples while they simply flow along microfluidic paths. The LoC device is equipped with a compact linear array detector to capture and generate a new conceptual type of a digital hologram in the space-time domain, named here as Space-Time Digital Hologram (STDH). The reported results show that the method is a promising diagnostic tool for optofluidic investigations of biological specimens.

Sequential treatment with epirubicin, oxaliplatin and 5FU (EOF) followed by docetaxel, oxaliplatin and 5FU (DOF) in patients with advanced gastric or gastroesophageal cancer: a single-institution experience.

PURPOSE: The aim of this study was to evaluate the activity and safety of epirubicin (EPI), oxaliplatin (l-OHP) and 5fluorouracil (5FU) (EOF) followed by docetaxel (D), l-OHP and 5FU (DOF) in patients with advanced gastric or gastroesophageal junction (GEJ) cancer. METHODS: Forty-five patients were enrolled: 26 gastric and 19 GEJ cancer. Median age was 69 years (range 34-83); ECOG performance status was 0-1 in 37 patients. Treatment consisted of EPI 50 mg/m(2) combined with l-OHP 130 mg/m(2) on day 1 and continuous infusion 5FU 750 mg/m(2) days 1-5 (EOF), every 3 weeks for a maximum of 4 cycles. After EOF completion, patients received D 70 mg/m(2) combined with l-OHP 130 mg/m(2) on day 1 and continuous infusion 5FU 750 mg/m(2) days 1-5 (DOF), every 3 weeks for a maximum of 4 cycles. RESULTS: After sequential EOF/DOF, the overall response rate was 51.1 % (95 % CI 35.7-66.2 %) and 93.3 % of patients were progression free 6 months after the onset of chemotherapy. The median progression-free survival was 9.5 months (95 % CI 8.0-11.9 months), and the median overall survival was 15.8 months (95 % CI 13.6-18.9 months). Grade 3 neutropenia was observed in 15 patients (33.3 %) after sequential EOF/DOF. CONCLUSIONS: The sequential treatment EOF/DOF is feasible in well-selected patients with advanced gastric or GEJ cancer and shows encouraging survival results.

Rechallenge of docetaxel combined with epirubicin given on a weekly schedule in advanced castration-resistant prostate cancer patients previously exposed to docetaxel and abiraterone acetate: a single-institution experience.

The aim of this paper was to evaluate the activity and tolerability of weekly docetaxel (D) combined with weekly epirubicin (EPI) in patients with advanced castrate-resistant prostate cancer (CRPC) previously exposed to D and abiraterone acetate (AA). Locally advanced or metastatic CRPC patients with 0-2 performance status, who had progressed after D and AA therapy, were included in the study. Previous treatment with chemotherapy agent cabazitaxel was also admitted. Treatment consisted of D 30 mg/m(2) intravenously (i.v.) and EPI 30 mg/m(2) i.v., every week (D/EPI). Chemotherapy was administered until disease progression or unacceptable toxicity. In our institution, twenty-six patients received D/EPI: their median age was 72 years (range 59-83 years). Twenty-three (88.5%) patients had bone metastases. A decrease in PSA levels ≥50% was observed in seven patients (26.9%, 95% CI: 0.11-0.47); of these, five had achieved a ≥50% PSA response during prior first-line D and six had achieved a PSA response during prior AA Among the subjects who were symptomatic at baseline, pain was reduced in nine patients (38.1%) with a significant decrease in analgesic use. Median progression-free survival was 4.4 months (95% CI, 3-5.2), and median overall survival was 10.7 months (95% CI, 8.9-18.4). Treatment was well tolerated and no grade 4 toxicities were observed. Our findings suggest that weekly D/EPI is feasible and active in heavily pretreated advanced CRPC patients and seem to support the hypothesis that the addition of EPI to D may lead to overcome the resistance to D in a subgroup of patients.

Spatio-temporal scanning modality for synthesizing interferograms and digital holograms.

We investigate the spatio-temporal scanning of a single-pixel row for building up synthetic interferograms or digital holograms, shifted each other of a desired phase step. This unusual recording modality exploits the object movement to synthesize interferograms with extended Field of View and improved noise contrast. We report the theoretical formulation of the synthetizing recording process and experimental evidence of various cases demonstrating quantitative phase retrieval by adopting this intrinsic phase-shifting procedure. The proposed method could be particularly suited in all cases where the object shift is an intrinsic feature of the investigated system, as e.g. in microfluidics imaging.

Premenstrual syndrome and beyond: lifestyle, nutrition, and personal facts.

AIM: The aim was to study prevalence of menstrual symptoms and their presenting patterns, and to assess nutrition and behaviours correlates. METHODS: The study enrolled 108 women, who were prospectively assessed for menstrual symptoms; a 212 items questionnaire about lifestyle and nutrition was administered. Diagnosis and intensity of symptoms were separately compared for dietary and other factors. The χ2 test or Fisher exact test as required and a multivariate analysis were performed. RESULTS: Seventeen percent of the sample reported no symptoms; core PMD was diagnosed in 14%, and related to symptoms of moderate intensity, to late menarche, irregular, long cycles, dysmenorrhoea, and to lower intake of complex carbohydrates, fibers, calcium; non-cyclical pattern in 27%, and related to symptoms of severe intensity, to young age, unemployed, to poor health, stress, traumatic events, abnormal BMI, to long, heavy menses, to lower intake of fibers, magnesium, vitamin B6, and to higher intake of fat, proteins, sodium, caffeine; variant PMD-premenstrual exacerbation in 42%, and related to symptoms of severe - moderate intensity, to young age, students, unemployed, sedentary work, abnormal BMI, stress, to cycle irregularities, heavy menses, to higher intake of simple sugars and sodium, and to lower intake of calcium, magnesium, and vitamin B6. Regardless of diagnosis, severe intensity of symptoms was associated with young age /students (P<0.001), lower education, unemployed, smoke, sedentary work, poor health, stress, heavy menses (P<0.01), dysmenorrhea, more unfavorable and less favorable nutrients. CONCLUSION: Lifestyle, nutrition, and general health considerations seem to be important issues in the management of menstrual symptoms, prospective symptoms chart being mandatory for diagnosis.

Spontaneous regression of low-grade cervical intraepithelial lesions is positively improved by topical bovine colostrum preparations (GINEDIE®). A multicentre, observational, italian pilot study.

OBJECTIVES: Human papillomavirus (HPV) is the causal agent of cervical cancer. The great majority of abnormal Pap test results - almost 90% - is referrable to either atypical squamous intraepithelial lesion or CIN1. For these lesions, worldwide agreement exists concerning the high rate - ranging from 40% to 70% - of spontaneous regression over a period of 1-5 years. Host's immune response is a key point influencing the natural history of these conditions. Bovine colostrum is a natural agent positively promoting several immune activities against bacterial and viral agents. The aim of this report was to evaluate the potential positive effect of bovine colostrum-containing vaginal tablets administered to CIN1 diagnosed patients in a prospective trial in regards to spontaneous regression rate. PATIENTS AND METHODS: A series of 256 consecutive patients with histologically proven CIN1 recruited in a multicentre, observational, Italian study. Patients have been enrolled in a 24-weeks protocol of treatment and re-tested at the end of the study. Rates of regression have been recorded. RESULTS: Overall regression rate to a negative histology at the end of the 6 month follow up was 75.5%. CONCLUSIONS: Regression to normal histology was observed in a very high rate of cases in a very short period compared to the natural history of these lesions. CIN1 patients could benefit from bovine colostrum topical administration in terms of significantly shortening the regression time.

Imaging through scattering microfluidic channels by digital holography for information recovery in lab on chip.

We tackle the problem of information recovery and imaging through scattering microfluidic chips by means of digital holography (DH). In many cases the chip can become opalescent due to residual deposits settling down the inner channel faces, biofilm formation, scattering particle uptake by the channel cladding or its damaging by corrosive substances, or even by condensing effect on the exterior channels walls. In these cases white-light imaging is severely degraded and no information is obtainable at all about the flowing samples. Here we investigate the problem of counting and estimating velocity of cells flowing inside a scattering chip. Moreover we propose and test a method based on the recording of multiple digital holograms to retrieve improved phase-contrast images despite the strong scattering effect. This method helps, thanks to DH, to recover information which, otherwise, would be completely lost.