Targeted and Untargeted Mass Spectrometry-Based Metabolomics for Chemical Profiling of Three Coffee Species.

While coffee beans have been studied for many years, researchers are showing a growing interest in coffee leaves and by-products, but little information is currently available on coffee species other than Coffea arabica and Coffea canephora. The aim of this work was to perform a targeted and untargeted metabolomics study on Coffea arabica, Coffea canephora and Coffea anthonyi. The application of the recent high-resolution mass spectrometry-based metabolomics tools allowed us to gain a clear overview of the main differences among the coffee species. The results showed that the leaves and fruits of Coffea anthonyi had a different metabolite profile when compared to the two other species. In Coffea anthonyi, caffeine levels were found in lower concentrations while caffeoylquinic acid and mangiferin-related compounds were found in higher concentrations. A large number of specialized metabolites can be found in Coffea anthonyi tissues, making this species a valid candidate for innovative healthcare products made with coffee extracts.

Coffee Leaves: An Upcoming Novel Food?

Unlike those of coffee beans, the healthy properties of coffee leaves have been overlooked for a long time, even if they are consumed as a beverage by local communities of several African countries. Due to the presence of xanthines, diterpenes, xanthones, and several other polyphenol derivatives as main secondary metabolites, coffee leaves might be useful to prevent many daily disorders. At the same time, as for all bioactive molecules, careless use of coffee leaf infusions may be unsafe due to their adverse effects, such as the excessive stimulant effects on the central nervous system or their interactions with other concomitantly administered drugs. Moreover, the presence of some toxic diterpene derivatives requires careful analytical controls on manufactured products made with coffee leaves. Accordingly, knowledge about the properties of coffee leaves needs to be increased to know if they might be considered a good source for producing new supplements. The purpose of the present review is to highlight the biosynthesis, metabolism, and distribution of the 4 main classes of secondary metabolites present in coffee leaves, their main pharmacological and toxicological aspects, and their main roles in planta. Differences in coffee leaf chemical composition depending on the coffee species will also be carefully considered.

Implementation of multispecies ecological networks at the regional scale: analysis and multi-temporal assessment.

Today, major landscape changes affect ecological connectivity exerting adverse effects on ecosystems. Connectivity is a critical element of landscape structure and supports ecosystem functionality. Landscape connectivity can be efficiently increased in landscape ecology by building ecological networks (EN) through models mimicking the interaction between animal and vegetal species and their environment. ENs are important in sustainable landscape planning, where they need to be studied both by applying landscape metrics and by performing multi-temporal analyses. This paper presents theoretical and practical evidence of an analysis of a multispecies ecological network in Calabria (Italy) and its changes over three decades. Landscape connectivity was modeled basing on 66 focal faunal species' requirements. Human disturbance (HD) was defined and assessed according to distance from different disturbance sources. This allowed for the definition of overall habitat quality (oHQ). Landscape permeability to the animal movement was focused as the main concept to measure landscape fragmentation. Landscape graph theory was applied to perform a spatial comparison of the ENs robustness. Many binary and probabilistic indices and landscape morphological spatial pattern analysis (MSPA) were used in this perspective. We obtained a set of ecological networks, including nodes, patches (i.e., habitat patches), linkages, and corridors, all intertwined in one giant component. The multi-temporal analysis showed many indices' stationary values, while MSPA yielded an increase of habitat quality and habitat patches in core areas. This methodological approach allowed for assessing the regional EN's robustness in the time-span considered, thus providing a reliable tool for landscape planners and communities.

Bilateral M1 anodal transcranial direct current stimulation in post traumatic chronic minimally conscious state: a pilot EEG-tDCS study.

OBJECTIVE: We tested the preliminary effects of bilateral anodal transcranial direct current stimulation (tDCS) in patients with disorders of consciousness. DESIGN: Open label pilot study. SUBJECTS: Ten chronic (greater than 12 months) patients in a minimally conscious state (MCS) following severe traumatic brain injury. METHODS: The patients received 10 sessions of bilateral M1 anodal tDCS. Behavioural changes were assessed with the Coma Recovery Scale-Revised (CRS-R) before stimulation (T-1, T0), after five sessions (T1), at the end of the stimulation (T2), after two weeks (T3) and after three months (T4). Moreover, an EEG assessment was conducted. RESULTS: Eight out of 10 patients showed new clinical signs of consciousness; specifically, a 2-point CRS-R improvement was detected in the last follow-up (p = 0.004). EEG upper α bandwidth was greater in the parietal site at T1 (p < 0.034). In addition, we found a significant correlation between behavioral and EEG indices at T1 (r =  0.89; p =  0.001). CONCLUSION: This preliminary study presents several limitations (small sample size and no control group). However, it provides important initial data that can be used to design randomized clinical trials testing this novel approach in MCS and to further explore EEG as a neural marker for the effects of tDCS.

Targeted metabolomics and transcript profiling of methyltransferases in three coffee species.

Coffee plants contain well-known xanthines as caffeine. Three Coffea species grown in a controlled greenhouse environment were the focus of this research. Coffea arabica and C. canephora are two first principal commercial species and commonly known as arabica and robusta, respectively. Originating in Central Africa, C. anthonyi is a novel species with small leaves. The xanthine metabolites in flower, fruit and leaf extracts were compared using both targeted and untargeted metabolomics approaches. We evaluated how the xanthine derivatives and FQA isomers relate to the expression of biosynthetic genes encoding N- and O-methyltransferases. Theobromine built up in leaves of C. anthonyi because caffeine biosynthesis was hindered in the absence of synthase gene expression. Despite this, green fruits expressed these genes and they produced caffeine. Given that C. anthonyi evolved successfully over time, these findings put into question the defensive role of caffeine in leaves. An overview of the histolocalisation of xanthines in the different flower parts of Coffea arabica was also provided. The gynoecium contained more theobromine than the flower buds or petals. This could be attributed to increased caffeine biosynthesis before fructification. The presence of theophylline and the absence of theobromine in the petals indicate that caffeine is catabolized more in the petals than in the gynoecium.

The Minimal assessment Protocol for Cerebral Stroke 2020 (PMIC2020): a multicenter feasibility study in post-stroke inpatient rehabilitation.

BACKGROUND: In 2008, a Working Group of the Italian Society of Physical and Rehabilitation Medicine (SIMFER) published the first minimum protocol for assessing stroke patients (PMIC) to define functional needs and outcomes. The recent PMIC revision (PMIC2020) introduces a document for all rehabilitation settings, incorporating updated measurement tools. AIM: The aim of this study was to investigate the PMIC2020 feasibility and administration time (AT) in post-stroke inpatients and to examine the influence of demographic and clinical variables on AT. DESIGN: Multicenter prospective observational study. SETTING: Eight Italian rehabilitation centers for post-acute inpatients. POPULATION: Adult patients consecutively admitted to rehabilitation after ischemic/hemorrhagic stroke, reporting the first event or recurrence, with a modified Barthel Index (mBI)<75 points, without cognitive impairment and clinical instability. METHODS: PMIC2020 was administered at admission (T0) and discharge (T1), recording AT of each section/ tool. A feasibility questionnaire was administered to assessors. Univariate and multivariate analyses were conducted to investigate the effect of demographics and clinical variables on AT. RESULTS: One hundred fifty-one subjects were enrolled at T0 and 139 at T1; the mean±SD AT (seconds) was 1634±401 at T0 and 1087±360 at T1 (P<0.001). National Institute of Health-Stroke Scale and Mini-Mental State Examination required the highest AT. All but two scales had significantly lower AT at T1 (P<0.05). Severe disability (as measured by mBI) was associated with higher AT than either complete or minimal/absent disability. The feasibility questionnaire showed good PMIC2020 appraisal by assessors without relevant critical issues. CONCLUSIONS: PMIC2020 was feasible in post-acute inpatient rehabilitation settings. No relevant critical issue was raised by users. Even though more comprehensive than PMIC, PMIC2020 required only slightly more AT (27 minutes at T0 and 18 minutes at T1, on average); more AT was needed to assess patients with severe disability. CLINICAL REHABILITATION IMPACT: The study has immediate transferability for the National Health Service, as PMIC2020 can be routinely implemented in clinical practice and research to assess stroke patients' needs and outcomes. The updated measures allow more immediate comparisons with international data on stroke rehabilitation. Future research should investigate the PMIC2020 feasibility in other rehabilitation settings and its relevance in predicting stroke rehabilitation needs and outcomes.

Redefining a minimal rehabilitation assessment protocol for severe acquired brain injuries.

BACKGROUND: The assessment of patients with severe Acquired Brain Injury (sABI) is mandatory in every phase and setting of care, and requires a multidimensional and interdisciplinary approach, to develop the individual rehabilitation project, and monitor long-term functional outcomes. In 2001 the Italian Society of Physical and Rehabilitation Medicine (SIMFER) published the minimal assessment protocol for traumatic sABI, providing a comprehensive, standardized functional assessment based on the International Classification of Functioning, Disability and Health (ICF), 2001. In 2007, a new protocol was published, extended to all sABI patients (PMGCA). In 2019, the SIMFER appointed a working group to provide a revised, updated version: the PMGCA2020. AIM: The purpose of this study was to describe the minimal assessment protocol to be applied at every stage and setting of the care process of patients with sABI. METHODS: The working group, including one neurologist and 11 physiatrists experts in sABI rehabilitation, performed a review of the international recommendations for sABI assessment focusing on the following key words: "sABI assessment," "sABI rehabilitation," "sABI prognostic factors," "sABI rehabilitation assessment," "sABI outcome," in MEDLINE. Revision and integration proposals by each member were written and motivated, discussed and voted. RESULTS: The PMGCA2020 is addressed to sABI adult patients. It investigates the main clinical problems of sABI at any time of the rehabilitation pathway. It includes a demographic/anamnestic section, a clinical/functional assessment section and an outcome measures section following the ICF model of functioning and the model of the construction of the rehabilitation project. CONCLUSIONS: The PMGCA2020 provides an updated tool for the multidimensional rehabilitation assessment of sABI patients, at any stage of the rehabilitation pathway. Further studies will allow the validation of this minimum set of variables paving the way to an assessment standardization of patients with sABI in the rehabilitation settings. CLINICAL REHABILITATION IMPACT: This minimum set of variables, defining patient's functioning and clinical status and outcomes, at every stage and setting of the care process to provide a framework for the standardization of the clinical evaluation of patients with sABI in rehabilitation settings.

Redefining a minimal assessment protocol for stroke rehabilitation: the new "Protocollo di Minima per l'ICtus" (PMIC2020).

The use of standardized assessment protocols is strongly recommended to identify patient's needs, outcomes, and predictors of response to specific interventions in Stroke rehabilitation. In 2008, the Italian Society of Physical and Rehabilitation Medicine (SIMFER) published the minimal protocol for the person with stroke ("Protocollo di Minima per l'ICtus" [PMIC]), in reference to the International classification of Functioning, Disability and Health. In 2019, the SIMFER appointed a working group to provide a revised, updated version in line with the most recent literature and suitable for all rehabilitation settings: the PMIC2020. Descriptive study based on the consensus of a panel of experts. The study setting includes all the rehabilitation settings. The population included is represented by stroke survivors with disability. The coordinator of the SIMFER National Stroke Section appointed the working group, including the 8 Stroke Section board members, and 4 more experts (3 physiatrists; 1 neurologist). An extensive revision of the international literature on stroke assessment recommendations was performed; each proposed change from PMIC was written and motivated, discussed and voted. The PMIC2020 is a single form, to be administered at any time of the rehabilitation pathway, including a minimum set of variables, consisting of a demographic/anamnestic section, and a clinical/functional assessment section. Newly introduced tools included measures of malnutrition (BMI<18.5); pain in verbal and non-verbal patients (Numeric Rating Scale-pain, Pain Assessment in Advanced Dementia Scale); neurological impairment (National Institute of Health-Stroke Scale); activity (Modified Barthel Index, Short Physical Performance Battery); and participation (Frenchay Activity Index). The PMIC2020 provides an updated tool for the multidimensional rehabilitation assessment of the stroke patient, at any stage of the rehabilitation pathway, aiming at a shared minimum set of variables defining patient's needs and at defined outcomes across different rehabilitation facilities and settings. The PMIC2020 identifies patient's needs, outcomes, and predictors of response to specific interventions in Stroke rehabilitation and provide ground for a highly needed Stroke Registry.

Hospitals with and without neurosurgery: a comparative study evaluating the outcome of patients with traumatic brain injury.

BACKGROUND: We leveraged the data of the international CREACTIVE consortium to investigate whether the outcome of traumatic brain injury (TBI) patients admitted to intensive care units (ICU) in hospitals without on-site neurosurgical capabilities (no-NSH) would differ had the same patients been admitted to ICUs in hospitals with neurosurgical capabilities (NSH). METHODS: The CREACTIVE observational study enrolled more than 8000 patients from 83 ICUs. Adult TBI patients admitted to no-NSH ICUs within 48 h of trauma were propensity-score matched 1:3 with patients admitted to NSH ICUs. The primary outcome was the 6-month extended Glasgow Outcome Scale (GOS-E), while secondary outcomes were ICU and hospital mortality. RESULTS: A total of 232 patients, less than 5% of the eligible cohort, were admitted to no-NSH ICUs. Each of them was matched to 3 NSH patients, leading to a study sample of 928 TBI patients where the no-NSH and NSH groups were well-balanced with respect to all of the variables included into the propensity score. Patients admitted to no-NSH ICUs experienced significantly higher ICU and in-hospital mortality. Compared to the matched NSH ICU admissions, their 6-month GOS-E scores showed a significantly higher prevalence of upper good recovery for cases with mild TBI and low expected mortality risk at admission, along with a progressively higher incidence of poor outcomes with increased TBI severity and mortality risk. CONCLUSIONS: In our study, centralization of TBI patients significantly impacted short- and long-term outcomes. For TBI patients admitted to no-NSH centers, our results suggest that the least critically ill can effectively be managed in centers without neurosurgical capabilities. Conversely, the most complex patients would benefit from being treated in high-volume, neuro-oriented ICUs.

First impact of COVID-19 on services and their preparation. "Instant paper from the field" on rehabilitation answers to the COVID-19 emergency.

This paper reports the immediate impact of the epidemic on rehabilitation services in Italy, the first country in Europe hit by COVID-19. In a country with almost 5000 Physical and Rehabilitation Medicine physicians, the webinar had 230 live viewers (4.5%), and more than 8900 individual visualizations of the recorded version. The overall inadequate preparation of the rehabilitation system to face a sudden epidemic was clear, and similar to that of the acute services. The original idea of confining the COVID-19 cases to some areas of rehabilitation wards and/or hospitals, preserving others, proved not to be feasible. Continuous reorganization and adaptation were required due to the rapid changes. Overall, rehabilitation needs had to surrender to the more acute emergency, with total conversion of beds, wards and even hospitals. The quarantine needs heavily involved also outpatient services that were mostly closed. Rehabilitation professionals needed support, but also acted properly, again similarly to what happened in the acute wards. The typical needs of rehabilitation, such as human and physical contacts, but also social interactions including patient, team, family and caregivers, appeared clearly in the current unavoidable need of being suppressed. These notes could serve the preparation of other services worldwide.

Early rehabilitation for severe acquired brain injury in intensive care unit: multicenter observational study.

BACKGROUND: The increased survival after a severe acquired brain injury (sABI) raise the problem of making most effective the treatments in Intensive Care Unit (ICU)/Neurointensive Care Unit (NICU), also integrating rehabilitation care. Despite previous studies reported that early mobilization in ICU was effective in preventing complications and reducing hospital stay, few studies addressed the rehabilitative management of sABI patients in ICU/NICU. AIM: To collect clinical and functional data about the early rehabilitative management of sABI patients during ICU/NICU stay. DESIGN: Prospective, observational, multicenter study. SETTING: Fourteen facilities supplied by intensive neurorehabilitation units and ICU/NICUs. POPULATION: Consecutive sABI patients admitted to ICU/NICU. METHODS: Patients were evaluated at admission and then every 3-5 days. Clinical, functional and rehabilitative data, including Glasgow Coma Scale (GCS), Disability Rating Scale (DRS), The Rancho Los Amigos Levels of Cognitive Functioning Scale (LCF), Early Rehabilitation Barthel Index (ERBI), Glasgow Outcome scale (GOS) and Functional Independence Measure (FIM) were collected. RESULTS: One hundred and two patients (F/M 44/58) were enrolled. The mean duration of ICU stay was 24.7±13.9 days and the first rehabilitative evaluation occurred after 8.7±8.8 days. Regular postural changes and multijoint mobilization were prescribed in 63.7% and 64.7% cases, respectively. The mean session duration was 38±11.5 minutes. Swallowing evaluation was performed in 14.7% patients, psychological support was provided to 12.7% of patients' caregivers, while 17.6% received a psycho-educational intervention, and 28.4% were involved in interdisciplinary team meetings. The main discharge destinations were Severe Acquired Brain Injury rehabilitation units for 43.7%, intensive neurorehabilitation units for 20.7%. CONCLUSIONS: Data showed that early rehabilitation was not diffusely performed in sABI subjects in ICU/NICU and rehabilitative interventions were variable; one-third of subjects were not referred to dedicated rehabilitation unit at discharge. CLINICAL REHABILITATION IMPACT: The study stresses the need to spread and implement a rehabilitative culture also for critical ill patients due to neurological diseases.