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Graphical abstract: From the fundamentals and the working principles of hybrid halide perovskite solar cells to their path to commercialization.

Cardiovascular (CV) diseases are burdened by high mortality and morbidity, being responsible for half of the deaths in Europe. Although hypertension is recognized as the most important CV risk factor, hypertension awareness, and blood pressure (BP) control are still unsatisfactory. In 2017 and 2018, respectively >10 000 and >5000 individuals took part in the May Measurement Month (MMM) campaign in Italy, of whom 30.6% and 26.3% were found to have high BP, respectively. To raise public awareness on the importance of hypertension and to collect BP data on a nation-wide scale in Italy. In the frame of the MMM campaign, an opportunistic cross-sectional survey of volunteers aged ≥18 years was carried out in May 2019. BP measurement, the definition of hypertension, and statistical analysis followed the standard MMM protocol. Screening was conducted in multiple sites by health personnel. Among the 10 182 people screened (females: 52.3%, mean age 58 ± 16years) mean BP was 127/78 mmHg, and 3171 (31.1%) participants had arterial hypertension, of whom 62.1% were aware of being hypertensive. Diabetes, body mass index >25 kg/m2were associated with higher BP and previous myocardial infarction with lower BP. For the third consecutive year we collected a nation-wide snapshot of BP control in a large sample of individuals. The high participation, with some yearly fluctuations likely due to the limitations of the sampling technique, confirms the power of this kind of health campaign in reaching a significant number of people to raise awareness on health topics.

From the fifteenth century onwards, devotional texts represented a prominent part of the output of the Italian printing press. Much of this production, which often represented a privileged way to access the biblical text, is still largely unexplored. My article will analyse a selection of devotional writings printed at the end of the fifteenth century and in the first three decades of the sixteenth century that were directed to a large audience of laymen and women of medium to low literacy. I will analyse how these texts entered the domestic devotional practices of Italian devotees, focussing especially on reading. I will take into account their suggestions about how, when, and by whom reading should be performed; what readings devotees were encouraged to pursue; how the ideal reader was shaped in the paratextual apparatuses; and, finally, what textual tools the readers were offered to perform their reading practices.

Aims Remodelling of the cardiovascular system (including heart and vasculature) is a dynamic process influenced by multiple physiological and pathological factors. We sought to understand whether remodelling in response to a stimulus, exercise training, altered with healthy ageing.Methods A total of 237 untrained healthy male and female subjects volunteering for their first time marathon were recruited. At baseline and after 6 months of unsupervised training, race completers underwent tests including 1.5T cardiac magnetic resonance, brachial and non-invasive central blood pressure assessment. For analysis, runners were divided by age into under or over 35 years (U35, O35).Results Injury and completion rates were similar among the groups; 138 runners (U35: n= 71, women 49%; O35: n= 67, women 51%) completed the race. On average, U35 were faster by 37 minutes (12%). Training induced a small increase in left ventricular mass in both groups (3 g/m2, P< 0.001), but U35 also increased ventricular cavity sizes (left ventricular end-diastolic volume (EDV)i +3%; left ventricular end-systolic volume (ESV)i +8%; right ventricular end-diastolic volume (EDV)i +4%; right ventricular end-systolic volume (ESV)i +5%; P< 0.01 for all). Systemic aortic compliance fell in the whole sample by 7% (P= 0.020) and, especially in O35, also systemic vascular resistance (–4% in the whole sample, P= 0.04) and blood pressure (systolic/diastolic, whole sample: brachial –4/–3 mmHg, central –4/–2 mmHg, all P< 0.001; O35: brachial –6/–3 mmHg, central –6/–4 mmHg, all P< 0.001).Conclusion Medium-term, unsupervised physical training in healthy sedentary individuals induces measurable remodelling of both heart and vasculature. This amount is age dependent, with predominant cardiac remodelling when younger and predominantly vascular remodelling when older.

Purpose: Inherited kidney diseases are among the leading causes of kidney failure in children, resulting in increased mortality, high healthcare costs and need for organ transplantation. Next-generation sequencing technologies can help in the diagnosis of rare monogenic conditions, allowing for optimized medical management and therapeutic choices. Methods: Clinical exome sequencing (CES) was performed on a cohort of 191 pediatric patients from a single institution, followed by Sanger sequencing to confirm identified variants and for family segregation studies. Results: All patients had a clinical diagnosis of kidney disease: the main disease categories were glomerular diseases (32.5%), ciliopathies (20.4%), CAKUT (17.8%), nephrolithiasis (11.5%) and tubular disease (10.5%). 7.3% of patients presented with other conditions. A conclusive genetic test, based on CES and Sanger validation, was obtained in 37.1% of patients. The highest detection rate was obtained for ciliopathies (74.4%), followed by nephrolithiasis (45.5%), tubular diseases (45%), while most glomerular diseases and CAKUT remained undiagnosed. Conclusions: Results indicate that genetic testing consistently used in the diagnostic workflow of children with chronic kidney disease can (i) confirm clinical diagnosis, (ii) provide early diagnosis in the case of inherited conditions, (iii) find the genetic cause of previously unrecognized diseases and (iv) tailor transplantation programs.

Several features of amyotrophic lateral sclerosis (ALS) impact on sexuality and intimate relationship; however, the issue has received poor attention so far. We performed a systematic literature review in order to provide an up-to-date account of sexuality in ALS. References were identified by searches of PubMed, Web of Science, Scopus and PsycINFO (1970–2017, English literature). The following were the key terms: ‘sexual’ OR ‘sexuality’ OR ‘intimacy’ OR ‘marital’ AND ‘ALS’ OR ‘Amyotrophic Lateral Sclerosis’ OR ‘Motor Neuron Disease’ OR ‘MND’. Titles and abstracts were screened for relevance and a full-text analysis was performed on the selected articles. Studies were included if they referred to sexual well-being/activities/functions or intimate relationship between patients and their partners and management of such topic by clinicians. Eligibility assessment was performed independently by two reviewers. A thematic and level of evidence classification of studies was performed. Studies’ design, objectives, measurements and outcomes were summarised. Thirty articles were included and four topics were identified: intimacy in the dyads; sexual activities in patients and with their partners; sexual function disturbances; and sexuality and cognitive-behavioural alterations. The quality of the studies varies, with globally poor level of evidence. Some sexuality issues have been only sparsely addressed, such as gender-related differences, same-sex relationships and sexual activities other than intercourse. Sexuality in ALS is still not adequately considered by clinicians and researchers. We present preliminary recommendations for improving sexuality and intimacy at any ALS multidisciplinary clinics.

Mammalian mRNA biogenesis requires specific recognition of a hexanucleotide AAUAAA motif in the polyadenylation signals (PAS) of precursor mRNA (pre-mRNA) transcripts by the cleavage and polyadenylation specificity factor (CPSF) complex. Here we present a 3.1-Å-resolution cryo-EM structure of a core CPSF module bound to the PAS hexamer motif. The structure reveals the molecular interactions responsible for base-specific recognition, providing a rationale for mechanistic differences between mammalian and yeast 3′ polyadenylation. The cryo-EM structure of the human core CPSF complex, containing CPSF160, WDR33, CPSF30 and Fip1 subunits, bound to its RNA target reveals the mechanism of PAS recognition.

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Background It has been previously shown in patients with heart failure that exercise-based rehabilitation programmes may improve functional capacity and autonomic response. The aim of this study was to investigate this issue further by evaluating whether an association exists between autonomic adaptations and improvements of aerobic capacity in a general population of coronary artery disease patients undergoing cardiac rehabilitation.Methods Ninety consecutive patients (age 60 ± 11 years) attended a rehabilitation programme of moderate continuous training (25 ± 8 sessions, 2–3 sessions/week). Functional capacity expressed as oxygen uptake (peak VO2) and autonomic function expressed as chronotropic response and heart rate recovery were evaluated by cardiopulmonary exercise tests before and after the rehabilitation programme. According to the expected mean increase in functional capacity, coronary artery disease patients were divided into two groups: those who improved peak VO2by more than 2.6 ml/kg/min (R group) and those who did not (NR group). Effects of the rehabilitation programme were compared in R and NR groups.Results The number and intensity of exercise sessions did not differ between R (N= 39) and NR (N= 51) groups. However, only R patients improved chronotropic response (R: from 45.1 ± 16.9% to 72.7 ± 34.1%, P< 0.01; NR: from 49.3 ± 18.6% to 48.2 ± 36.5%, P= NS) and heart rate recovery (R: from 16.9 ± 7.0 bpm to 21.0 ± 8.7 bpm, P< 0.01; NR: from 15.2 ± 9.9 bpm to 15.8 ± 8.5 bpm, P= NS). After training both chronotropic response and heart rate recovery were significantly higher in R than NR patients.Conclusions The improvement in aerobic capacity of coronary artery disease patients following exercise-based cardiac rehabilitation programmes is associated with positive adaptations of autonomic function.

The objective of this study is to simulate tool wear in drilling of nickel-based alloys, in particular Inconel 718. When machining these kind of materials, the impact of the thermal and mechanical phenomena generated by tool wear on the surface integrity is of prime concern. For this reason, it important to study the influence of tool wear on tool life, on final part quality and on cutting force and power consumption. Tool wear is caused by several phenomena (adhesion, abrasion, erosion, diffusion, corrosion, fracture etc.) depending on selected cutting parameters (cutting velocity, feed rate, etc.). In some cases these wear mechanisms can be described by analytical models which are function of physical quantities involved in process (temperature, pressure and sliding velocity along the cutting surface). Usually, commercial FEM software allows to implement these tool wear models but without tool geometry update. To overcoming this limitation, a suitable subroutine considering tool geometry update was developed and implemented in SFTC DEFORM-3D FEA software to simulate tool wear in drilling of Inconel 718. A good agreement was obtained between the predicted and measured tool wear data.

The management of articular fractures is always a matter of concern. Each articular fracture is different from the other, whatever the classification system used and the surgical or non-surgical indications employed by the surgeon. The main goals remain anatomical reduction, stable fixation, loose body removal and minimal invasiveness.Open procedures are a compromise. Unfortunately, it is not always possible to meet every treatment goal perfectly, since associated lesions can pass unnoticed or delay treatment, and even in a ‘best-case’ scenario there can be complications in the long term.In the last few decades, arthroscopic joint surgery has undergone an exponential evolution, expanding its application in the trauma field with the development of arthroscopic and arthroscopically-assisted reduction and internal fixation (ARIF) techniques. The main advantages are an accurate diagnosis of the fracture and associated soft-tissue involvement, the potential for concomitant treatments, anatomical reduction and minimal invasiveness. ARIF techniques have been applied to treat fractures affecting several joints: shoulder, elbow, wrist, hip, knee and ankle.The purpose of this paper is to provide a review of the most recent literature concerning arthroscopic and arthroscopically-assisted reduction and internal fixation for articular and peri-articular fractures of the upper limb, to analyse the results and suggest the best clinical applications.ARIF is an approach with excellent results in treating upper-limb articular and peri-articular fractures; it can be used in every joint and allows treatment of both the bony structure and soft-tissues.Post-operative outcomes are generally good or excellent. While under some circumstances ARIF is better than a conventional approach, the results are still beneficial due to the consistent range of movement recovery and shorter rehabilitation time.The main limitation of this technique is the steep learning curve, but investing in ARIF reduces intra-operative morbidity, surgical errors, operative times and costs.Cite this article: Dei Giudici L, Faini A, Garro L, Tucciarone A, Gigante A. Arthroscopic management of articular and peri-articular fractures of the upper limb. EFORT Open Rev2016;1:325-331. DOI: 10.1302/2058-5241.1.160016.

Nocturnal periodic breathing occurs more frequently in men than in women with various clinical and pathophysiologic conditions. The mechanisms accounting for this sex-related difference are not completely understood. Acetazolamide effectively counteracts nocturnal periodic breathing, but it has been investigated almost exclusively in men. Our aim was to explore possible determinants of nocturnal periodic breathing in a high-altitude setting both in men and in women. We hypothesized that increased hypoxic chemosensitivity in men could be associated with the development of nocturnal periodic breathing at altitude more frequently than in women, and that acetazolamide, by leftward shifting the CO2ventilatory response, could improve nocturnal periodic breathing at altitude in a sex-independent manner.

Locomotion exists in diverse forms in nature; however, little is known about how closely related species with similar neuronal circuitry can evolve different navigational strategies to explore their environments. Here, we investigate this question by comparing divergent swimming pattern in larval Danionella cerebrum(DC) and zebrafish (ZF). We show that DC displays long continuous swimming events when compared with the short burst-and-glide swimming in ZF. We reveal that mesencephalic locomotion maintenance neurons in the midbrain are sufficient to cause this increased swimming. Moreover, we propose that the availability of dissolved oxygen and timing of swim bladder inflation drive the observed differences in the swim pattern. Our findings uncover the neural substrate underlying the evolutionary divergence of locomotion and its adaptation to their environmental constraints.

The five subunits of transcription factor NF-κB have distinct biological functions. NF-κB signaling is important for skin homeostasis and aging, but the contribution of individual subunits to normal skin biology and disease is unclear. Immunohistochemical analysis of the p50 and c-Rel subunits within lesional psoriatic and systemic sclerosis skin revealed abnormal epidermal expression patterns, compared with healthy skin, but RelA distribution was unaltered. The skin of Nfkb1−/−and c-Rel−/−mice is structurally normal, but epidermal thickness and proliferation are significantly reduced, compared with wild-type mice. We show that the primary defect in both Nfkb1−/−and c-Rel−/−mice is within keratinocytes that display reduced proliferation both in vitroand in vivo. However, both genotypes can respond to proliferative stress, with 12-O-tetradecanoylphorbol-13-acetate–induced epidermal hyperproliferation and closure rates of full-thickness skin wounds being equivalent to those of wild-type controls. In a model of bleomycin-induced skin fibrosis, Nfkb1−/−and c-Rel−/−mice displayed opposite phenotypes, with c-Rel−/−mice being protected and Nfkb1−/−developing more fibrosis than wild-type mice. Taken together, our data reveal a role for p50 and c-Rel in regulating epidermal proliferation and homeostasis and a profibrogenic role for c-Rel in the skin, and identify a link between epidermal c-Rel expression and systemic sclerosis. Modulating the actions of these subunits could be beneficial for treating hyperproliferative or fibrogenic diseases of the skin.

We determine in high anisotropy Pt/Co- and Ni/Co-based multilayer materials, which are candidates for high density storage applications, the contributions to the extraordinary Hall effect. In order to gain more insight into the physics underlying these processes, the dependence of the extraordinary Hall resistivity on the longitudinal resistivity is used to identify the skew scattering and the combined intrinsic and side jump effects when analyzing their variation with temperature. Furthermore, the influence of an additional amorphous CoFeB layer is shown, revealing a possibility to enhance the extraordinary Hall effect using this material.

Iron is tightly connected to oxygen homeostasis and erythropoiesis. Our aim was to better understand how hypoxia regulates iron acquisition for erythropoiesis in humans, a topic relevant to common hypoxia-related disorders. Forty-seven healthy volunteers participated in the HIGHCARE project. Blood samples were collected at sea level and after acute and chronic exposure to high altitude (3400-5400 m above sea level). We investigated the modifications in hematocrit, serum iron indices, erythropoietin, markers of erythropoietic activity, interleukin-6, and serum hepcidin. Hepcidin decreased within 40 hours after acute hypoxia exposure (P < .05) at 3400 m, reaching the lowest level at 5400 m (80% reduction). Erythropoietin significantly increased (P < .001) within 16 hours after hypoxia exposure followed by a marked erythropoietic response supported by the increased iron supply. Growth differentiation factor-15 progressively increased during the study period. Serum ferritin showed a very rapid decrease, suggesting the existence of hypoxia-dependent mechanism(s) regulating storage iron mobilization. The strong correlation between serum ferritin and hepcidin at each point during the study indicates that iron itself or the kinetics of iron use in response to hypoxia may signal hepcidin down-regulation. The combined and significant changes in other variables probably contribute to the suppression of hepcidin in this setting.

Little is known about the effects of cardiovascular drugs at high altitude.

Background:Immunophenotypic modulation is induced by steroids in pediatric Bcell precursor acute lymphoblastic leukemia BCPALL patients during remission induction therapy.Methods:We cultured BCPALL blasts from diagnostic bone marrow BM samples n 20 in the presence of prednisone on stroma layer obtained from BMderived mesenchymal cells to maintain viability. Antigen expression was assessed by multiparametric flow cytometry.Results:Leukemia samples that sustained the treatment in vitro with prednisone, showed significative reduction of CD10 and CD34 expression compared with control, and it was comparable with that observed in residual leukemic cells of the same patients in BM at day 15 of treatment. Modulated cells were viable as determined by Annexin V staining and preserved light scattering properties. Of note, the extent of antigen modulation in vitro correlated with response to prednisone in vivo.Conclusions:The prednisoneinduced immunophenotypic modulation can be reproduced in vitro and this phenomenon may reflect sensitivity to chemotherapy. © 2008 Clinical Cytometry Society

Background:Hemodialysis (HD) is associated with cardiovascular structural modifications; moreover, during HD, rapid electrolytic changes occur. Both factors may favor the onset of atrial fibrillation. Methods:To define the prevalence of atrial fibrillation and identify associated factors, 488 patients on long-term HD therapy (age, 66.6 ± 13.4 years; men, 58.0%; duration of HD, 76.5 ± 84.3 months) were studied. Results:Atrial fibrillation was reported in 27.0% of patients; paroxysmal in 3.5%, persistent in 9.6%, and permanent in 13.9%. Clinical and echocardiographic variables were considered: patients with atrial fibrillation were older (71.8 ± 9.3 versus 64.7 ± 14.2 years; P< 0.01), and its prevalence increased with age. Patients with arrhythmia had a longer duration of dialysis therapy (93.2 ± 100.5 versus 70.2 ± 76.7 months; P= 0.02). Atrial fibrillation was associated significantly with ischemic heart disease (P< 0.01), dilated cardiomyopathy (P< 0.01), acute pulmonary edema (P< 0.05), valvular disease (P< 0.05), cerebrovascular accidents (P< 0.05), and predialytic hyperkalemia (P< 0.05). Patients with atrial fibrillation more frequently showed left atrial dilatation (59.8% versus 34.5%; P< 0.0001), and in these subjects, left ventricular ejection fraction was significantly lower (53.9% versus 57.4%; P= 0.029). No association was found between arrhythmia and hypertension or diabetes. Multivariate analysis confirmed that patient age (P< 0.001), duration of HD therapy (P= 0.001), and left atrial dilatation (P< 0.001) were associated with atrial fibrillation. Conclusion:Atrial fibrillation is much more frequent in HD patients than in the general population; age, duration of HD history, presence of some heart diseases, and left atrial dilatation are associated with the arrhythmia.

Spin injection is involved in many experiments in the field of spin electronics. Examples of spin injection effects can be found in several types of transport experiments in metallic structures, in the problem of spin injection from a ferromagnetic metal into a semiconductor or also in the reversal of magnetization by transverse spin injection, as reviewed in the oral presentation by one of us (AF) at ICMFS. Here we will limit our presentation to a general description of the physics of spin injection and to an illustration of this physics by experiments in which the spin accumulation generated by spin injection in metallic structures is directly detected by voltage measurements.

He⁺ ion irradiation is an excellent tool to modify the magnitude and direction of the exchange bias field on the sub-micrometer scale without affecting the sample topography. This effect has been utilized to magnetically pattern NiFe/FeMn exchange bias double layers using resist masks patterned by electron beam lithography. Ion irradiation through the masks leads to a local modification of the magnetization reversal behavior and allows to study the magnetization reversal as a function of the exchange bias field strength and the pattern dimensions on a single sample. Results are presented on the macroscopic and microscopic magnetization reversal using the magneto-optic Kerr effect and Lorentz microscopy.

Carnosol 1 , rosmanol 2 , carnosic acid 3 and 20-deoxocarnosol 4 , the main phenolic abietanes present in aerial parts of the Chilean medicinal plant Sphacele salviae were tested for antioxidant activity of measuring the decay of the radical cation diphenyl-picrylhydrazyl (DPPH). All compounds displayed higher antioxidant levels than BHT under the same conditions and carnosic acid was more efficient than vitamin E.

A chemical study of aerial parts of Haplopappus illinitus afforded five labdane diterpenoids whose structures were elucidated by high field NMR spectroscopy and 2D techniques. Four of them are dicarboxylic acids previously reported as the corresponding dimethyl ester derivatives and the fifth is a new compound.

The phagodepression activity of five furanocoumarins (FC), bergapten, xanthotoxin, psoralen, imperatorin, and angelicin, has been studied against larvae of Egyptian cotton leafworm Spodoptera littoralis(Boisduval) (Lepidoptera: Noctuidae) using a leaf-disk choice bioassay. The dose range used was 0–10 μg/cm2for linear furocoumarins and 0–30 μg/cm2for angelicin, the angular furanocoumarin. Dose–feeding deterrency activity correlations were governed by various sigmoidal functions, except in the case of imperatorin and bergapten, which had dose–response curves showing irregular traces with two maxima. All five FCs induced various degrees of phagodepression in S. littoralis; at 3 μg/cm2, the relative feeding deterrence was bergapten = xanthotoxin > psoralen = imperatorin = angelicin. Structure–activity correlations indicated that a methoxy group on C-5 or C-8 enhanced the activity. Comparison of experimental feeding deterrence of binary mixtures of imperatorin with xanthotoxin, bergapten, or psoralen at various relative concentrations with the calculated additive activity of each combination indicated that a proportion of 40% or more of imperatorin may exert a greater antifeedant effect on S. littoralisthan the sum of individual compounds. The effect of angelicin also was examined in combination with psoralen, both of which are present in Psoraleaplants. Their mixtures yielded a clear synergistic effect in the 20–80% angelicin range (coactivity coefficient = 124–133). By contrast, the effect of angelicin on xanthotoxin was only additive. In the dose–response curves of imperatorin/bergapten combinations, synergism was found at >60% imperatorin relative concentration and 1 μg/cm2, whereas lower proportions led to antagonism. The threshold between the two opposing effects was found to depend on the total FC concentration employed. Some natural systems contain FCs in the range of synergistic proportions recorded here and, thus, may have been produced by the host to increase its defensive effect at a lower metabolic cost.

We study the magneto-transport in vertical quantum dots boththeoretically and experimentally. We focus our attention onthe non-linear transport regime. We demonstrate that the peakamplitudes in the I–V characteristics show a dramaticallydifferent behaviour as a function of the magnetic field,depending on the value of the angular momentum of the dotstate through which tunnelling occurs. The investigationallows us to probe details of the quantum dot wavefunctionsand to distinguish tunnelling through localised states relatedto impurities or to quantum dots.

We have studied the effects of magnetic field on the fluctuations of the electron density of states in a disordered conductor, a contact in the resonant tunnelling structure GaAlAs/GaAs/GaAlAs. The local density of states (LDOS) in the contact is detected by an impurity which acts as a ‘spectrometer’. In the regime of weak magnetic field, ωcτ ≤ 1, a linear increase of the fluctuation magnitude with B2 is seen, due to the decrease of the volume where the LDOS is formed and a decrease in the statistical averaging. In the regime ωcτ ≥ 1, large oscillations of the fluctuation magnitude have been observed. We suggest an explanation of this new effect, de Haas-van Alphen type oscillations in the fluctuation magnitude. The behaviour of impurity spectrometers in a magnetic field and images given by a complex spectrometer are also discussed.

Highly anisotropic sub-100nm InP patterns presenting low roughness have been obtained using SiCl"4 reactive ion etching. Aspect ratios above 20 are reported with low sidewall damage characterized by transmission electron microscopy and conductance measurements. The influence of the material used as an etch mask on the anisotropy and the roughness of the etched structures is investigated.

This contribution presents an application of scanning near-field optical microscopy to the characterization of semi-conductors. It is based on the photocurrent mapping of a patterned Au/GaAs structure (Schottky barrier) under local illumination by the nanosource. The results obtained with different wavelengths, metallized or dielectric probes and different bias voltages exhibit photocurrent variations independent ofthe topography and induced by interface defects. Finally, from this study of a patterned planar structure, we propose a method to determine the mean free pathof the charge carriers in the volume.

Workers' remittances represent a sizeable component of international trade flows in goods and services. The paper tries to assess to what extent workers remittances are responsive to key macroeconomic variables. We first develop a simple model of altruistic transfers and show that the real exchange rate may play a crucial role in affecting the remittance behaviour of migrants. Econometric estimation of a remittance equation for a sample of five Mediterranean countries indicates that the real exchange rate is indeed a significant determinant of remittances. Further support in this respect comes from an analysis of remittance behaviour by foreign workers in Germany. We also find strong evidence to support the claim that remittances are altruistically motivated, as indicated by the systematically negative coefficient associated with recipients' income.

In order to measure differences in the pollen growth rate of numerous lines of maize and to investigate the main features of their genetic control, gametophyte growth was studied in vitro and in vivo.