Fast liquid chromatography-tandem mass spectrometry method for the simultaneous determination of phytocannabinoids in oily based preparations.

The reported method aims to be a powerful aid for the simultaneous determination of tetrahydrocannabinol (THC), cannabidiol (CBD), cannabinol (CBN), cannabigerol (CBG), tetrahydrocannabinolic acid (THCA), cannabidiolic acid (CBDA), and tetrahydrocannabivarin (THCV) in oily based preparations. The chromatographic separation was carried out using an Hypersil Gold PFP (50 × 2.1 mm, 1.9 µm) column, using H2O + 2 mM ammonium formate + 0.2 % formic acid (M1) and Methanol + 2 mM ammonium formate + 0.2 % formic acid (M2) as mobile phases. The flow rate was set 0.4 mL/min. Specifically, this method was validated in terms of linearity, limit of detections and quantifications (LODs and LOQs), accuracy (precision and trueness, both intra and interday), selectivity, and matrix effects. This procedure allowed quantifying seven phytocannabinoids in less than 10 min. The validated method shows a good linearity within the range 0.25-1000 ng/mL, while precision and trueness (intra- and inter-day) were below <13.25 % and 7.59 %, respectively. Regarding the matrix effect, the method satisfies all the requirements, except for the THC and THCV, where it reaches about 120 %. This element does not affect the method performances as it has been observed that this value is constant and reproducible and therefore does not involve errors in the quantitative analysis. The method was tested and applied on more 70 different oily based preparations. Furthermore, starting from four different cannabis cultivar (FM2, Bedrolite, Bedrocan, and Bediol), it allowed to evaluate the reproducibility of the magistrali preparations. The real samples, in fact, derive from different local pharmacies, and were analyzed by the accredited UNI CEI EN ISO/IEC 17025:2018, Pharmatoxicology Laboratory (ACCREDIA, lab n. 2274 ASLPE, accreditation number 1822 L), accordingly to the current regulations.

Body mass index, lifestyles, physical performance and cognitive decline: the "Treviso Longeva (TRELONG)" study.

OBJECTIVES: The relative contributions of risk factors, as body mass index (BMI), depression, chronic diseases, smoking, and lifestyles (as physical and performance activity, social contacts and reading habit) to cognitive decline in the elderly are unclear. We explored these variables in relation to 7-year cognitive decline in long-lived Italian elderly. DESIGN: Secondary data analysis of a longitudinal study of a representative, age-stratified, population sample. SETTING: The TREVISO LONGEVA (TRELONG) Study, in Treviso, Italy. PARTICIPANTS: 120 men and 189 women, age 77 years and older (mean age 80.2 ± 6.9 years) survivors after seven years of follow up. MEASUREMENTS: Cognitive decline measured as difference between Mini-Mental State Examination (MMSE) score in 2003 and in 2010; Body mass index (BMI), handgrip, Short Physical Performance Battery (SPPB) score, social contacts, reading habit, sight, hearing, schooling, mediterranean diet and multiple clinical and survey data recorded at baseline in 2003. RESULTS: In separate univariate analyses, age, SPPB score < 5, depressive symptoms (GDS) and more comorbidities (CCI) were associated with greater cognitive decline. Otherwise higher BMI, higher handgrip, reading habit, non-deteriorated sight and hearing, and schooling were protective. In a final multivariate model, age and higher BMI were associated with greater cognitive decline while reading habits was protective. SPPB score < 5 tends, though weakly, to be associated with greater cognitive decline. These associations remained with multivariate adjustment for gender, schooling, Charlson co-morbidity index (CCI) and baseline MMSE. CONCLUSION: Age and higher baseline BMI, independent of gender, and other confounding factors, are risk factors for cognitive decline. Reading habit plays a protective role seven years later among northern Italian adults aged 70 years or older. Low physical performance tends, though weakly, to be associated with greater cognitive decline.

The Treviso Dementia (TREDEM) Study: A Biomedical, Neuroradiological, Neuropsychological and Social Investigation of Dementia in North-Eastern Italy.

BACKGROUND: The incidence of dementia increases exponentially with age but knowledge of real disease-modifying interventions is still limited. OBJECTIVES: To describe the study design and methods of a large prospective cohort study aimed at exploring the complex underlying relationships existing among cognition, frailty, and health-related events in older persons with cognitive impairment. DESIGN: Prospective cohort study of a representative population of outpatients attending the Treviso Cognitive Impairment Center between 2000 and 2010. SETTING: The TREVISO DEMENTIA (TREDEM) Study conducted in Treviso, Italy. PARTICIPANTS: 490 men and 874 women, mean age 79.1 ± 7.8 years (range 40.2-100 years). MEASUREMENTS: Physiological data, biochemical parameters, clinical conditions, neuroradiological parameters (e.g., brain atrophy and cerebral vascular lesions identified by computerized tomography scans), neuropsychological assessment, and physical function markers were measured at baseline. Patients were followed-up to 10 years. RESULTS: The final sample included in the study was predominantly composed of women and characterized by an initial physical function impairment and increased vascular risk profile. Cognitive function of the sample population showed moderate cognitive impairment (Mini Mental State Examination 20.2 ± 6.3; Clinical Dementia Rating 1.2 ± 0.7), and a prevalence of vascular dementia of 26.9%. Cortical, subcortical and hippocampus atrophy were all significantly correlated with age and cognitive function. CONCLUSION: Results obtained from the preliminary analyses conducted in the TREDEM study suggest that the database will support the accomplishment of important goals in understanding the nature of cognitive frailty and neurodegenerative diseases.

Factors related to disability: evidence from the "Treviso Longeva (TRELONG) study".

Prolongation of life is an important public health goal as long as there is an emphasis on the quality of life (QoL) and independent living. Diminishing abilities to ambulate and participate in activities of daily living point to a serious decline in functional health, increasing the risk of institutionalization and death. In our work we found a pattern of factors associated with disability, especially cognitive impairment, as well as stroke, physical activity and performance, reading, and the nutritional biomarkers, blood albumin and high-density lipoprotein cholesterol (HDL-C). The attention to this cluster of markers, suggesting multidimensional prevention, may have unexpected good effects against disability.

The Treviso Longeva (Trelong) study: a biomedical, demographic, economic and social investigation on people 70 years and over in a typical town of North-East of Italy.

Longevity is a complex process resulting from genetic and environmental factors, as well as their interaction. These factors are poorly understood, and the comparison among health status, socio-economics, demographic and other characteristics of the elderly people can help in understanding these complex interactions. Such an interdisciplinary approach is necessary to allow an appropriate evaluation of longevity. Here we report the methodology and the first results of a representative study performed in 2003-2004 on people of 70 years and over, living in a typical town of North-East of Italy. In the research we collected biomedical, demographic, socio-economic and quality of life (QoL) data.

Associations of the plasma interleukin 6 (IL-6) levels with disability and mortality in the elderly in the Treviso Longeva (Trelong) study.

IL-6 expression is regulated by the interplay of several transcriptional and hormonal factors, including sex steroids and glucocorticoids. In late life IL-6 expression increases as a result from loss of the normally inhibiting sex steroids. IL-6 is one of several proinflammatory cytokines. It has been proposed that many chronic inflammatory diseases are the result of a dysregulation of IL-6 expression. In this work we demonstrate that increased IL-6 levels in elderly are associated with higher disability and mortality, also independently of age and comorbidity.

Homocysteine in Alzheimer disease and vascular dementia.

Hyperhomocysteinemia is an important risk factor for atherosclerosis and it has recently been suggested as a diagnostic marker for Alzheimer disease (AD). The present studies compared homocysteinemia in patients with AD and with vascular dementia (VD),as well as in controls. Homocysteinemia was measured in 137 probable AD patients,diagnosed by the NINCDS-ADRDA criteria, in 40 probably VD patients diagnosed by the NINDS-AIREN criteria, and in 42 control subjects. Homocysteine levels were significantly higher in AD and VD groups, than in controls, however, VD patients were significantly older than the controls. The proportion of females was higher in the AD group, while serum folate and B12 vitamin levels tended to be lower in both the AD and VD groups, compared to the controls. In order to adjust the results for these potentially interfering factors, a multivariate ANCOVA calculation was performed, where homocysteine levels were analyzed considering 2 grouping factors (gender and neurological type) and 4 covariates (age, albumin,serum folate and vitamin B12 ). This analysis confirmed that even after adjusting for the covariates, the difference between AD and VD groups and controls remain highly significant,while neither the gender itself, nor the interaction of gender and the neurological type had any significant influence on the homocysteine levels. The main finding, therefore, is a significant increase of homocysteine levels in the 2 disease groups, compared to controls.

Linear accelerator radiosurgery of three-dimensional irregular targets.

Current radiosurgical techniques allow concentration of radiation from external sources into spherical targets. Nonspherical target volumes require a modification of the irradiation technique. Moreover, isodose shaping is sometimes necessary to spare nearby important radiosensitive structures. To meet these prerequisites we have introduced some technical refinements: (1) computer-controlled rectilinear translations of the target in combination with different angular positions of the source and (2) computer-controlled rotations of the target around a vertical axis in combination with different angular positions of the source. The first solution has proven efficient for treating irregular targets, while the second one was employed for focusing the radiation dose inside volumes obtained by rotation of ellipses.

New technique for three-dimensional linear accelerator radiosurgery.

A new method for external stereotactic focal irradiation of three-dimensional irregular target volumes is proposed. In this method the target is irradiated by a linear accelerator set in various angular positions around the isocenter. During the irradiation the target is translated in a direction perpendicular to the beam. By controlling the velocity of the translation it is possible to modify the configuration of therapeutic isodoses so as to make them follow the borders of the target.

Axiomatization of genetics. 1. Biological meaning.

Axiomatization is a trend in science to settle and reduce fundamental assertions, from which all the others are deducible. Classical genetics has been extensively axiomatized and formalized by Woodger (1952), who resorted to 53 axioms and 12 theorems. The molecular settlement of modern genetics provides the basis for a different axiomatic theory. In this paper 3 axioms and 14 theorems are informally expressed. They cover a few elementary laws of "chromosome genetics" in Eukaryotes. The biological problems connected with the further development of this first attempt are discussed.

Axiomatization of genetics 2. Formal development.

When certain assumptions are claimed to "found" a scientific theory, two kinds of controls are needed: it must be proved that every assertion of the theory is derived from the assumptions and that no assumption depends on (that is, derives from) the other ones. The formal approach (when possible) is the best method of achieving these results. In this paper the axiomatization of the elementary laws of genetics considered in Rizzotti & Zanardo (1986) is translated into a formal language and an axiomatic theory is defined in which the (translated) laws are deducible. Independence results are also considered.

External stereotactic irradiation by linear accelerator.

Stereotactic radiotherapy has two advantages: (a) the possibility of giving high radiation doses to small but spatially well-defined target volumes and (b) the presence of a stepped dose gradient between the target volume and the surrounding healthy tissues. To utilize these advantages, the authors built a new stereotactic head frame by which the intracranial target is fixed to the rotational isocenter of a 4-MV linear accelerator. The collimator openings are selected according to the volume and the three-dimensional configuration of the target, and the radiation dose is based on the radiosensitivity of the lesion. After the patient is fixed to the frame, the radiation source and the patient are rotated so that the target is irradiated through infinite portals distributed over the convexity of the skull. It is thereby possible to obtain very high radiation doses centered into the target with a stepped dose gradient. The preliminary radiodosimetric tests and the operative technique are described. The advantages of this technique compared to interstitial radiotherapy and Leksell's radiosurgery are emphasized. This noninvasive procedure has been used to treat a series of intracranial tumors.

Stereotactic radiosurgery utilizing a linear accelerator.

The authors have developed a radiosurgical technique based on multiple arc irradiations. The target is fixed to the rotational isocenter of a Varian 4 MV linear accelerator. The first irradiation is carried out while the radiating source is rotating on a 100-140 degrees arc. The patient is then rotated around a vertical axis passing through the target, and arc irradiations are repeated in different angular positions. By this technique it is possible to obtain very steep dose gradients at the borders of the target volume. High doses are usually delivered in two shots. 47 patients have been treated so far in a clinical trial that started in November 1982. The paper deals with the preliminary results (more than 6 months' follow-up) obtained in patients affected by nonresectable brain tumors and AVMs.

[Stereotaxic irradiation with a linear accelerator]. / L'irradiazione stereotassica con acceleratore lineare.

At least two fundamental requirements must subsist for every stereotaxic radiotherapy treatment: 1) a capacity to administer extremely high doses of radiation to small areas which have been precisely defined in three dimensions; 2) the presence of a steep gradient between adjacent isodose curves so that damage to the tissues close to the target may be reduced to the minimum. The authors have built a special stereotaxic device for the fixing of the head of the patient (together with the chosen intracranial target), once the indispensable neuroradiological evaluations and bioptic controls have been effected, at the isocenter of a linear accelerator (Varian Clinac 4). Once the dimensions of the collimator have been chosen in accordance with the volume and the three dimensional conformation of the target and the dose to be administered has been decided upon in accordance with the histological nature of the lesion, both the linear accelerator and the patient are rotated about the isocenter of the therapy unit. It this way it is possible to reach extremely high dosages with very steep isodose gradients, centered exclusively upon the target ("gamma knife"). The procedure employed is described in full. The dosimetric evaluations which preceded the use of the set-up in clinical practice are illustrated as well. The advantages that this technique offers when compared with interstitial brachytherapy and/or Leksell's radiosurgical procedures are stressed. The described procedure, which is entirely bloodless and thus applicable also to high vascularized lesions, has so far been used in a limited series of cases.

A new method for utilizing CT data in stereotactic surgery: measurement and transformation technique.

The proposed system is essentially based on the definition of a coordinate system, rigidly connected with the skull, by the introduction of three reference X-ray opaque points, secured into the bone of the skull. Target and reference points CT coordinates are given by the image display console. The three reference points are also identified on stereoradiographs, and their actual stereotactic coordinates are calculated by a geometrical approach which takes into account radiological distortion. A programmed computer links the target, identified on tomograms, to the three points system and calculates its stereotactic coordinates.