Immunomodulatory effects of metronomic vinorelbine (mVRL), with or without metronomic capecitabine (mCAPE), in hormone receptor positive (HR+)/HER2-negative metastatic breast cancer (MBC) patients: final results of the exploratory phase 2 Victor-5 study.

Tregs are able of suppressing tumor-specific effector cells, such as lymphocytes CD8+, CD4+ and Natural Killer cells. Different drugs, especially different schedules of administration, like metronomic chemotherapy (mCHT), seem to be able to increase anticancer immunity, by acting on downregulation of Tregs. Most of the data available regarding the immunomodulating effect of mCHT have been obtained with Cyclophosphamide (CTX). Aim of the present study was to explore the effects of mVRL and mCAPE administration, alone or in combination, on T cells. Observation of 13 metastatic breast cancer patients lasted controlling for 56 days, where Treg frequencies and function, spontaneous anti-tumor T-cell responses were monitored, as well as the clinical outcome. No depletion in Treg absolute numbers, or percentage of T lymphocytes, was observed. Only in 5 patients, a modest and transient depletion of Tregs was observed during the first 14 days of treatment. To better describe the effect on Tregs, we subsequently looked at the variations in Memory, Naïve and Activated Treg subpopulations: we observed a trend in reduction for memory Treg (Treg MEM) and an increase for Treg Naïve (Treg NAIVE) and Treg Activated (Treg ACT) components. We finally analyzed the average trend of Treg in the Treg depleted patients and non-depleted ones, without fiding any significant differences. The trend of the Treg MEM appeared different, showing a reduction during the first 14 days, followed by an increase at the levels before treatment at Day 56 in the group of depleted patients and a progressive substantial reduction in the group of non-depleted patients along the entire course of treatment. Opposed to the data known, treatment with mVRL w/o mCAPE did not show any effect on Tregs.

Error estimation on extracorporeal trajectory determination from body scans.

This study explores the magnitude of two sources of error that are introduced when extracorporeal bullet trajectories are based on post-mortem computed tomography (PMCT) and/or surface scanning of a body. The first source of error is caused by an altered gravitational pull on soft tissue, which is introduced when a body is scanned in another position than it had when hit. The second source of error is introduced when scanned images are translated into a virtual representation of the victim's body. To study the combined magnitude of these errors, virtual shooting trajectories with known vertical angles through five "victims" (live test persons) were simulated. The positions of the simulated wounds on the bodies were marked, with the victims in upright positions. Next, the victims were scanned in supine position, using 3D surface scanning, similar to a body's position when scanned during a PMCT. Seven experts, used to working with 3D data, were asked to determine the bullet trajectories based on the virtual representations of the bodies. The errors between the known and determined trajectories were analysed and discussed. The results of this study give a feel for the magnitude of the introduced errors and can be used to reconstruct actual shooting incidents using PMCT data.

Practical application of synthetic head models in real ballistic cases.

In shooting crimes, ballistics tests are often recommended in order to reproduce the wound characteristics of the involved persons. For this purpose, several "simulants" can be used. However, despite the efforts in the research of "surrogates" in the field of forensic ballistic, the development of synthetic models needs still to be improved through a validation process based on specific real caseworks. This study has been triggered by the findings observed during the autopsy performed on two victims killed in the same shooting incident, with similar wounding characteristics; namely two retained head shots with ricochet against the interior wall of the skull; both projectiles have been recovered during the autopsies after migration in the brain parenchyma. The thickness of the different tissues and structures along the bullets trajectories as well as the incident angles between the bullets paths and the skull walls have been measured and reproduced during the assemblage of the synthetic head models. Two different types of models ("open shape" and "spherical") have been assembled using leather, polyurethane and gelatine to simulate respectively skin, bone and soft tissues. Six shots have been performed in total. The results of the models have been compared to the findings of post-mortem computed tomography (PMCT) and the autopsy findings.Out of the six shots, two perforated the models and four were retained. When the projectile was retained, the use of both models allowed reproducing the wounds characteristics observed on both victims in terms of penetration and ricochet behaviour. However, the projectiles recovered from the models showed less deformation than the bullets collected during the autopsies. The "open shape" model allowed a better controlling on the shooting parameters than the "spherical" model. Finally, the difference in bullet deformation could be caused by the choice of the bone simulant, which might under-represent either the strength or the density of the human bone. In our opinion, it would be worth to develop a new, more representative material for ballistic which simulates the human bone.

Circulating tumor DNA changes for early monitoring of anti-PD1 immunotherapy: a proof-of-concept study.

BACKGROUND: Recent clinical results support the use of new immune checkpoint blockers (ICB), such as anti-PD-1 (e.g. nivolumab and pembrolizumab) and anti-PD-L1 antibodies. Radiological evaluation of ICB efficacy during therapy is challenging due to tumor immune infiltration. Changes of circulating tumor DNA (ctDNA) levels during therapy could be a promising tool for very accurate monitoring of treatment efficacy, but data are lacking with ICB. PATIENTS AND METHODS: This prospective pilot study was conducted in patients with nonsmall cell lung cancer, uveal melanoma, or microsatellite-instable colorectal cancer treated by nivolumab or pembrolizumab monotherapy at Institut Curie. ctDNA levels were assessed at baseline and after 8 weeks (w8) by bidirectional pyrophosphorolysis-activated polymerization, droplet digital PCR or next-generation sequencing depending on the mutation type. Radiological evaluation of efficacy of treatment was carried out by using immune-related response criteria. RESULTS: ctDNA was detected at baseline in 10 out of 15 patients. At w8, a significant correlation (r = 0.86; P = 0.002) was observed between synchronous changes in ctDNA levels and tumor size. Patients in whom ctDNA levels became undetectable at w8 presented a marked and lasting response to therapy. ctDNA detection at w8 was also a significant prognostic factor in terms of progression-free survival (hazard ratio = 10.2; 95% confidence interval 2.5-41, P < 0.001) and overall survival (hazard ratio = 15; 95% confidence interval 2.5-94.9, P = 0.004). CONCLUSION: This proof-of-principle study is the first to demonstrate that quantitative ctDNA monitoring is a valuable tool to assess tumor response in patients treated with anti-PD-1 drugs.

Bullet fragmentation preceding a contour shot: case study and experimental simulation.

In medico-legal literature, only a small number of publications deal with lethal injuries caused by shots with modified guns. This might lead to the conclusion that such cases are extremely rare. However, there are cases again and yet again. During the investigation process, the modified gun is of particular importance since it can show an unusual ballistic behaviour. The present paper reports on a suicide of a 60-year-old man, committed with a modified revolver and a lead bullet. The man had a single gunshot wound with entrance at the right temporal bone. Autopsy revealed that the bullet had fragmented into two major parts. The smaller one stood outside the cranial cavity and pushed its way alongside between the cranial bone and scalp to its end position in the left temporal area. The bigger part entered the cranial cavity and ended in the left parietal lobe. In shots on ballistic soap and on a head-model, the ballistics of the weapon and lead bullet were characterized. The angle necessary for bullet fragmentation was determined by shots on ballistic soap and turned out to be 55°-60° at a velocity of around 200 m/s. This knowledge was transferred to contact shots on a head-model consisting of a layered polyurethane sphere filled with 10 % ballistic gelatine and covered with a skin-like cap almost all around. The resulting injury pattern corresponded to the one of the suicide person. The bigger bullet part entered the skull while the smaller part pushed its way alongside between skin and skull causing an outer contour shot. Furthermore, the revolver was documented firing off two bullets by one trigger pull-a phenomenon of importance for forensic casework the authors have not found reported in forensic literature.

Expression of TIR8 receptor in chicken tissues.

The orphan receptor TIR8, also known as SIGIRR, belongs to the TLR/IL-1R (TIR) superfamily and plays an important role in the immune response. The signalling pathways of the receptors belonging to the TIR family are tightly regulated at multiple levels and through different mechanisms. TIR8 negatively modulates innate immunity and inflammatory responses in the areas where it is primarily expressed (gastrointestinal tract, kidney and lung). TIR8 has been well characterized in mouse, humans and in other Mammalian species, but it is still poorly known in chicken. Given the importance of gastrointestinal diseases in chicken, the aim of our study was to investigate the distribution of TIR8 in a wide panel of non-pathologic tissues and organs. TIR8 expression was analyzed in chicken samples at both levels of transcript mRNA and translated protein. The pattern of expression of TIR8 (ubiquitous) was similar to Mammals for some tissues (high levels in kidney and gastrointestinal tract), but it resulted unique for other tissues. High expression was detected in liver, pancreas and female reproductive tract. Interestingly, the receptor was highly expressed also in heterophils, the most common granulocytes of birds. Few isoforms of chicken TIR8 were detected by Western blot, suggesting the occurrence of different post-translational processing in different organs. Immunohistochemistry revealed TIR8 immunolabelling in chicken intestine and thymus. These results demonstrate that the receptor, although evolutionarily conserved, show species-specific peculiarities.

The innate immunity receptor TIR8/SIGIRR is expressed in the early developmental stages of chicken embryos.

The orphan receptor TIR8, also known as SIGIRR (Single Immunoglobulin IL-1R-Related molecule), belongs to the IL-1R/TLR (TIR) superfamily and plays an important role in the inflammatory responses. The signaling pathways of the receptors belonging to the TIR family are tightly regulated by both extracellular and intracellular mechanisms. TIR8 does not activate the transcription factors NFkB (nuclear factor kB) and IRF3 (interferon regulatory factor 3), although it negatively modulates the inflammatory responses. It acts as an antagonist for the IL-1 receptor family and triggers a negative pathway of the Toll-like/IL-1 receptor system, crucial for dampening inflammation stimuli in the gastrointestinal (GI) tract and in other organs (e.g. lung and kidney). The recent findings of TLRs expression in ovary and embryos of different species (mammals and chickens) are very important for an understanding of reproductive physiology and transovarian pathogen transmission. TIR8 was well characterized in mouse, humans and in other mammalian species, but it is still poorly characterized in the chicken. When TIR8 expression was measured in selected organs of chicken embryos of both broiler and layer types at different time points a unique pattern of expression was observed. Interestingly, TIR8 was detected during the first stages of chicken development (day 1 of incubation), and reached a remarkable level of expression by day 10. We observed this receptor to be ubiquitously expressed in the kidney, GI tract, Bursa of Fabricius, with the highest expression levels in liver and kidney. This pattern was comparable to those observed in post-hatching chickens and in mammals examined to date. No expression differences were observed between the two different chicken breeds (layer- and broiler-type) in the first incubation period (8 days). Whereas in some organs starting from day 10, higher TIR8 expression was observed in broiler-type compared to layer-type. These are the first findings concerning TIR8 expression in developmental stages and therefore they are of comparative value.

Extracorporeal bullet trajectory determination from scanned phantoms with bullet defects.

Shots with two different calibres (0.32 Auto and 9 mm Luger) were fired through phantoms that simulated human torsos, mounted on undercarriages with witness panels. The perforated phantoms were scanned with computed tomography (Siemens) using 80 kV and 140 kV and a slice thickness of 1 mm. The intracorporeal trajectories in the phantoms were compared to the known extracorporeal trajectories, derived from the perforations in witness panels. The discrepancy between the intracorporeal and extracorporeal trajectories, denoted as the absolute angle, was calculated for the trajectories before (front) and after (rear) the phantoms. Mean absolute angles at the front were lower than at the rear (2.27° vs. 4.54°) and the difference was statistically significant (p < 0.001). The results of the study imply that the line between the entrance and the exit wound in a scanned victim can be extended to the extracorporeal bullet trajectory leading towards the entrance wound. The absolute angles presented in this study give an impression of the expected errors with the two calibres. This can be helpful in shooting investigations to assess the position of the shooter from entrance and exit wounds in a scanned victim.

The influence of impact velocity on bullet trajectory deflection through ballistic gelatine.

This paper presents the results of a study on bullet trajectory deflection, for 9 mm Luger Full Metal Jacket Round Nose (FMJ-RN) bullets fired through 23-24 cm of ballistic gelatine. The bullets were fired at different velocities. Impact velocity, energy transfer and bullet trajectory deflection after gelatine perforation were measured and calculated. As was expected, energy transfer to the gelatine blocks generally increased with increasing impact velocity, indicating an altering bullet/gelatine interaction with altering velocity. This alteration did not result in a discernible alteration of bullet trajectory deflection. Deflection angles fell between 5.7° and 7.4° for 136 of the 140 fired shots, with four outliers below 5.7°.

Gene expression of pregnancy-associated glycoproteins-1 (PAG-1), interferon-tau (IFNt) and interferon stimulated genes (ISGs) as diagnostic and prognostic markers of maternal-fetal cellular interaction in buffalo cows.

The aim of this study was to determine the presence of Pregnancy-associated glycoprotein -1 (PAG-1) mRNA expression in the maternal circulation of pregnant buffaloes during the early stage of pregnancy. Contemporaneously, the mRNA expression levels of Interferon-tau (IFNt) and some Interferon stimulated genes (ISGs) (interferon stimulated gene 15 ubiquitin-like modifier interferon, ISG15; Mixoviruses resistance 1 and 2, MX1 and MX2; 2',5'-oligoadenylate synthase 1,OAS1) were evaluated in order to expand our knowledge of the molecular processes involved in the early stages of pregnancy and to identify potential biomarkers of maternal-fetal cellular interaction in buffalo. The study was conducted on 38 synchronized and artificially inseminated buffalo cows (d 0), divided ex post into 3 groups: Pregnant (n = 17), Non-pregnant (n = 15) and Embryo mortality (n = 6). Blood samples were collected on d 14, 19, 28 and 40 after artificial insemination (AI) for peripheral blood mononuclear cells (PBMCs) isolation. Expression levels of mRNA of PAG-1, IFNt, ISG15. MX1, MX2 and OAS1 were measured using RT-qPCR. No significant changes were observed in IFNt and PAG gene expressions between groups, while significant differences (p < 0.001) were found for ISG15, MX1, MX2, and OAS1. Pairwise comparisons revealed that the differences between groups occurred on days 19 and 28 post-AI. ISG15 proved to have the best diagnostic performance for distinguishing between pregnant animals and animals that experienced embryo mortality with the ROC analysis. According to the results of the univariate analyses, day 19 was identified as the most indicative to discriminate between groups while the most reliable genes for this differentiation were ISG15, MX1 and MX2. MX2 proved to be the best gene for discriminating pregnant buffaloes using the discriminant analysis, while MX1 was the gene that best predicted embryo mortality. Our results showed that among PAG-1, IFNt and ISGs expression as diagnostic and prognostic markers of maternal-fetal cellular interaction in buffalo cows, ISGs proved to be the best peripheral biomarkers for predicting pregnancy and embryonic mortality during the peri-implantation period. These insights into the mechanisms behind maternal-fetal interaction and the development of a method for the early detection of embryo distress may enable us to implement effective strategies to support embryo survival.

Translational and transdisciplinary approach to the human papilloma virus - Preliminary evidence from the Italian "HPV board: a future without papilloma virus" project.

Human papillomavirus (HPV) is considered to be one of the viral infections associated with cancers and other diseases. HPV is detected asymptomatically in the oral mucosa. The presence of human papillomavirus in the oral mucosa appears to be closely associated with a series of benign and malign oral lesions. The aim of this paper is to report the Italian experience in applying translational protocols, using new technologies and multidisciplinary strategies in Human Papilloma virus detection and treatment. The "HPV board: a future without papilloma virus" project was born, promoted by CNEL (Italian Council of Economics and Labor) with the collaboration of numerous scientific societies to commonly approach to public knowledge of HPV-related oral lesions and their clinical management. The preliminary results are related to the assessment of the proof-of-concept of this new project. More in details, "HPV Board" is a project that plans the presence of a working group, made up of otolaryngologists, dentists, oral and maxillofacial surgeons, in close contact with gynecologists, oncologists and pediatricians; this working group manages to combine very transversal skills, in order to promote primary prevention projects, early diagnosis and adequate therapies. The "HPV BOARD" project will give the opportunity to increase the attention of patients and doctors on the early diagnosis of oncological diseases dependent on infection by the infectious agent HPV. In this panorama, dentists will have the role of "first sentinel" of public health because oral health is an indicator, too often overlooked, for the prevention of numerous diseases.

Aerosol-generating procedures in head and neck surgery - can we improve practice after COVID-19?

The COVID-19 pandemic has had a dramatic impact on international medicine practice. The propensity for head and neck surgery to generate aerosols needs special consideration over and above simply adopting personal protective equipment. This study sought to interrogate the literature and evaluate whether which additional measures might provide benefit if routinely adopted in minimising viral transmission.

Videoconferencing for multidisciplinary team meetings in the coronavirus disease era - human factors awareness and recognition.

BACKGROUND: The coronavirus disease 2019 pandemic has led to the birth of videoconference multidisciplinary teams, which are now commonplace. This remote way of deciding care demands a new set of rules to ensure the quality of the complex decisions that are made for the patient group needing multidisciplinary care. Videoconference multidisciplinary teams bring with them novel forms of distraction that are under-appreciated and can impair decision-making. METHOD: A practical checklist was generated as applied to videoconference multidisciplinary teams using the principles of human factors awareness and recognition. RESULTS: Some of the strategies that should be adopted to minimise errors arising from human factors are: information technology support, a suitable environment to dial in, a global checklist employed prior to the videoconference, visible participants, avoiding distractions from other sources (e.g. e-mail, mobile phone), a videoconference sign-out and rapid dissemination of the outcomes sheet. CONCLUSION: This article presents a framework that uses human factors principles applied in this setting, which will contribute to enhanced patient safety, team working and a reduction in medical errors.

Influence of bullet type and muzzle-to-target distance on trajectory deflection through a soft tissue simulant.

The influence of two different variables on deflection was studied with 9mm Luger calibre bullets, fired through ballistic gelatine. To test whether bullet type affects trajectory deflection, bullets of four different types were fired (20 shots per type) through 25cm of 10% gelatine at a 5m muzzle-to target-distance. A Kruskal-Wallis test showed that the deflection of at least one bullet type was significantly different (p<.001, significance level of 0.05) from the deflection of the other bullet types. To test whether muzzle-to target-distance affects trajectory deflection, additional shots were fired with one of the four bullet types (7.5g FMJ-RN) at 1 and 15m muzzle-to target distances (20 shots per distance), again through 25 cm of 10% gelatine. A Kruskal-Wallis test showed that there was no significant difference (p=.72, significance level of 0.05) in deflection between the three muzzle-to target distances of 1, 5 and 15m.

Staphylococcus aureus intra-mammary infection affects the expression pattern of IL-R8 in goat.

IL-1R8 is a member of Interleukin-1 receptor family acting as a negative regulator of inflammation reliant on ILRs and TLRs activation. IL-1R8 role has never been evaluated in acute bacterial mastitis. We first investigated IL-1R8 sequence conservation among different species and its pattern of expression in a wide panel of organs from healthy goats. Then, modulation of IL-1R8 during natural and experimental mammary infection was evaluated and compared in blood, milk and mammary tissues from healthy and Staphylococcus aureus infected goats. IL-1R8 has a highly conserved sequence among vertebrates. Goat IL-1R8 was ubiquitously expressed in epithelial and lymphoid tissues with highest levels in pancreas. IL-1R8 was down-regulated in epithelial mammary cells following S. aureus infection. Interestingly it was up-regulated in leukocytes infiltrating the infected mammary tissues suggesting that it could represent a target of S. aureus immune evasion.

Rifle bullet deflection through a soft tissue simulant.

Trajectory deflections of 5.56 NATO and 7.62×39mm rifle bullets, fired through 7.5, 15 and 22.5cm of gelatine, were studied. The magnitude of the deflections from the bullets' original trajectories after perforation are related to the length and the profile of the (wound) channels through gelatine. After 7.5cm of penetration depth, deflection was less than 1°. With the longer channel lengths, bullet instability set in and subsequently, deflection was much larger. Deflection was highest with fragmented 5.56 NATO bullets after perforating 22.5cm of gelatine. The data from this study can be used to assess the degree of bullet deflection in trajectory reconstructions after incidents where human bodies were perforated with rifle bullets of the respective calibres and cartridge types.

A novel dressing for the combined delivery of platelet lysate and vancomycin hydrochloride to chronic skin ulcers: Hyaluronic acid particles in alginate matrices.

The aim of the present work was to develop a medication allowing for the combined delivery of platelet lysate (PL) and an anti-infective model drug, vancomycin hydrochloride (VCM), to chronic skin ulcers. A simple method was set up for the preparation of hyaluronic acid (HA) core-shell particles, loaded with PL and coated with calcium alginate, embedded in a VCM containing alginate matrix. Two different CaCl2 concentrations were investigated to allow for HA/PL core-shell particle formation. The resulting dressings were characterized for mechanical and hydration properties and tested in vitro (on fibroblasts) and ex-vivo (on skin biopsies) for biological activity. They were found of sufficient mechanical strength to withstand packaging and handling stress and able to absorb a high amount of wound exudate and to form a protective gel on the lesion area. The CaCl2 concentration used for shell formation did not affect VCM release from the alginate matrix, but strongly modified the release of PGFAB (chosen as representative of growth factors present in PL) from HA particles. In vitro and ex vivo tests provided sufficient proof of concept of the ability of dressings to improve skin ulcers healing.

Pistol bullet deflection through soft tissue simulants.

Trajectory deflections of pistol bullets from four different firearms, fired through soft tissue simulants under two different incidence and exit angles were studied. The data from this study can be used in reconstructions of shooting incidents where human soft tissues (not bones) were perforated with pistol bullets and assumptions must be made about bullet deflection in order to correctly reconstruct trajectories. The results demonstrate that deflection was influenced by the length of the "wound channel" through the simulants. In short, deflection was small to virtually absent with 5 and 10cm channel lengths. With channel lengths of 15, 20 and 25cm, there was a clear increase in deflection and/or a more erratic deflection behaviour with most shots. The data also suggest an influence of the angle of incidence and/or exit on both the direction and the magnitude of the deflection.

Illicit drugs and pharmaceuticals in swimming pool waters.

The occurrence of illicit drugs (cocaine, opioids, amphetamines and cannabis derivatives), some of their metabolites and 48 pharmaceuticals, was investigated in pool and source waters in ten Italian indoor swimming pools. The samples were analyzed by highperformance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), after solid phase extraction (SPE). Cocaine and its metabolites were found in nine swimming pools, at concentrations from 0.3 to 4.2 ng/L for cocaine, 1.1 to 48.7 ng/L for norcocaine, 0.7 to 21.4 ng/L for benzoylecgonine and 0.1 to 7.3 ng/L for norbenzoylecgonine. Opioids, amphetamines and cannabis derivatives were never detected. The most frequent pharmaceuticals were anti-inflammatory drugs: ibuprofen was found in all the pool waters, with a maximum 197 ng/L and ketoprofen was detected in 9/10 samples (maximum 127 ng/L). Among anticonvulsants, carbamazepine and its metabolite, 10,11-dihydro-10,11dihydroxycarbamazepine, were frequent in swimming pool water (8/10 samples) at concentrations up to 62 ng/L. The cardiovascular drug valsartan was also found frequently (8/10 samples), but at lower concentrations (up to 9 ng/L). Other pharmaceuticals were detected occasionally and at lower concentrations (atenolol, enalapril, paracetamol, hydroclorothiazide, irbesartan and dehydro-erythromycin). Carbamazepine, irbesartan and dehydroerythromycin were detected at very low levels (up to 5 ng/L) in only one of the four source water samples. A quantitative risk assessment showed that the health risk for humans to these substance in swimming pool waters was generally negligible, even for vulnerable subpopulations such as children and adolescents.