Prescribing Patterns of Psychotropic Drugs and Risk of Violent Behavior: A Prospective, Multicenter Study in Italy.

BACKGROUND: This prospective cohort study aimed at evaluating patterns of polypharmacy and aggressive and violent behavior during a 1-year follow-up in patients with severe mental disorders. METHODS: A total of 340 patients (125 inpatients from residential facilities and 215 outpatients) were evaluated at baseline with the Structured Clinical Interview for DSM-IV Axis I and II, Brief Psychiatric Rating Scale, Specific Levels of Functioning scale, Brown-Goodwin Lifetime History of Aggression, Buss-Durkee Hostility Inventory, Barratt Impulsiveness Scale, and State-Trait Anger Expression Inventory-2. Aggressive behavior was rated every 15 days with the Modified Overt Aggression Scale and treatment compliance with the Medication Adherence Rating Scale. RESULTS: The whole sample was prescribed mainly antipsychotics with high levels of polypharmacy. Clozapine prescription and higher compliance were associated with lower levels of aggressive and violent behavior. Patients with a history of violence who took clozapine were prescribed the highest number of drugs. The patterns of cumulative Modified Overt Aggression Scale mean scores of patients taking clozapine (n = 46), other antipsychotics (n = 257), and no antipsychotics (n = 37) were significantly different (P = .001). Patients taking clozapine showed a time trend at 1-year follow-up (24 evaluations) indicating a significantly lower level of aggressive behavior. Patient higher compliance was also associated with lower Modified Overt Aggression Scale ratings during the 1-year follow-up. CONCLUSION: Both inpatients and outpatients showed high levels of polypharmacy. Clozapine prescription was associated with lower Modified Overt Aggression Scale ratings compared with any other antipsychotics or other psychotropic drugs. Higher compliance was associated with lower levels of aggressive and violent behavior.

High resolution melting analysis for the identification of novel mutations in DKC1 and TERT genes in patients with dyskeratosis congenita.

Dyskeratosis congenita (DC) is a rare inherited bone-marrow failure syndrome with high clinical heterogeneity. Cells derived from DC patients present short telomeres at early ages, as a result of mutations in genes encoding components of the telomerase complex (DKC1, TERC, TERT, NHP2 and NOP10), or the shelterin complex (TINF2). However, mutations have been identified only in around 50% of the cases, indicating that other genes could be involved in the development of this disease. Indeed, mutations in TCBA1 or chromosome segment C16orf57 have been described recently. We have used HRM technology to perform genetic analysis in the above mentioned genes, in Spanish patients showing both, some clinical features of DC and short telomeres. The mutations have been identified by PCR amplification of DC genes followed by high resolution melting (HRM) and direct DNA sequencing analysis. We have identified seven new families with DC, three with X-linked DC and four with autosomal dominant DC, in which we have found two novel mutations in DKC1 (p.His68Arg and p.Lys390del) and four novel mutations in TERT gene (p.Pro530Leu, p.Arg698Trp, p.Arg971His and p.Arg698Gln). The results show that the use of HRM analysis enables a rapid and inexpensive identification of mutations in dyskeratosis congenita associated genes.

Effects of nutrient restriction on the metabolic profile of Bos indicus-influenced and B. taurus suckled beef cows.

Recent research from our group demonstrated that Bos indicus-influenced suckled beef cows had greater resilience to withstand nutrient restriction and establish pregnancy compared with B. taurus cows exposed to the same conditions. To further understand these findings, differences in metabolic profile between these same B. indicus-influenced and B. taurus females were explored. Suckled beef cows (n = 134) were enrolled in a completely randomized design with a 2 × 2 factorial arrangement of treatments. On day -21, Angus (AN; Bos taurus) and Brangus (BN; B. indicus-influenced) cows were randomly assigned to 1) a diet that met daily energy maintenance requirements (MAINT), or 2) a diet that restricted intake to 70% of the daily energy maintenance requirements (RESTR). Cows were exposed to an estrus synchronization protocol and received an embryo 7 d after ovulation was pharmacologically induced on day 0. Blood samples were collected on days -21 and 19 to determine circulating concentrations of non-esterified fatty acids (NEFA), ß-hydroxybutyrate (BHB), insulin, glucose, and IGF-1. Pregnancy status after embryo transfer was determined on day 28. As a consequence of the proposed diets, cows in the RESTR diet had less body condition score (BCS) on day 19 (P = 0.008) across breed types. Moreover, BCS change from day -21 to 19 was included as independent covariate into subsequent analyses, allowing for the comparison of breed types under an equivalent level of body reserve mobilization. A breed × diet interaction was observed for plasma insulin (P = 0.03) and IGF-1 (P = 0.04) on day 19, where AN-RESTR cows had less plasma concentrations on day 19 compared with AN-MAINT cows. Diets did not impact (P > 0.10) plasma insulin and IGF-1 concentrations in BN cows. No diet or breed effects were observed in circulating concentrations of NEFA, BHB, and glucose (P > 0.10). Across breed types and nutritional treatment, there was positive linear effect (P ≤ 0.04) of plasma concentrations of insulin and IGF-1 on the probability of pregnancy to fixed-time embryo transfer. In summary, the negative impacts of nutrient restriction on the somatotropic axis, independently of body tissue mobilization, were heightened in Bos taurus females compared with B. indicus-influenced cohorts, which corroborate with the differences observed in fertility between these subspecies.

Estrus synchronization and fixed-time artificial insemination alter calving distribution in Bos indicus influenced beef heifers.

To determine the effects of estrus synchronization (ES) and fixed-time artificial insemination (TAI) on calving distribution in Bos indicus influenced heifers, 751 Bos taurus × Bos indicus beef heifers were enrolled in a completely randomized design at 2 locations from January to May of 2016. Within location, all heifers were randomly assigned to one of two treatments: 1) SYNCH (n = 371); heifers were exposed to the 5-day CO-Synch + CIDR protocol where they were treated with 100 µg of GnRH, 25 mg of PGF2α, and a controlled internal drug releasing insert (CIDR) on d 0; heifers received 50 mg of PGF2α at CIDR removal on d 5, and were treated with 100 µg of GnRH and TAI 66 ± 2 h later on d 8; or 2) CONTROL (n = 380); heifers were exposed to natural service without ES or TAI. On d 9, all heifers were exposed to bulls for the remainder of the breeding season at each location. Blood samples were collected on d -9 and on d 0 to determine pretreatment estrous cyclicity (progesterone ≥ 1.0 ng/mL). Pregnancy was diagnosed via transrectal ultrasonography 54 d after TAI by determining the presence of a viable fetus. Fetal age was estimated based on fetal size and structural features at the time of pregnancy diagnosis. Pregnancy rates on d 54 differed (P < 0.001) between locations, but did not differ (P = 0.777) between CONTROL and SYNCH treatments. Pregnancy rates on d 54 were greater (P < 0.001) in cycling compared with non-cycling heifers (63.9 vs 42.4%). A greater (P < 0.05) proportion of SYNCH heifers became pregnant in the first 19 d of the breeding season compared with CONTROL heifers (52.2 vs 46.4%). Overall breeding season pregnancy rates did not differ (P = 0.982) between treatments. In summary, ES and TAI increased the percentage of heifers that conceived in the first 19 d of the breeding season, and therefore, potentially altered the calving distribution by ensuring that more heifers calve early during the subsequent calving season.

Expression of a lipid transfer protein in Escherichia coli and its phosphorylation by a membrane-bound calcium-dependent protein kinase.

Ha-AP10 is a basic antifungal peptide from sunflower seeds (Helianthus annuus antifungal peptide of 10 kDa) belonging to the family of plant lipid transfer proteins. We report here its expression in E. coli [Glutathione S-transferase (GST) system] and its phosphorylation by endogenous membrane-bound calcium-dependent protein kinases.

On the mechanical characteristics of a self-setting calcium phosphate cement.

OBJECTIVE: To perform a mechanical characterization of a self-setting calcium phosphate cement in function of the immersion time in Ringer solution. MATERIALS AND METHODS: Specimens of self-setting calcium phosphate cement were prepared from pure α-TCP powder. The residual strains developed during hardening stage were monitored using an embedded fiber Bragg grating sensor. Additionally, the evolution of the elastic modulus was obtained for the same time period by conducting low-load indentation tests. Micro-computed tomography as well as microscope-assisted inspections were employed to evaluate the porosity in the specimens. Moreover, diametral compression tests were conducted in wet and dried specimens to characterize the material strength. RESULTS: The volume of the estimated porosity and absorbed fluid mass, during the first few minutes of the material's exposure in a wet environment, coincide. The immersion in Ringer solution lead to a noticeable increase in the moduli values. The critical value of stresses obtained from the diametral compression tests were combined with the data from uniaxial compression tests, to suggest a Mohr-Coulomb failure criterion. CONCLUSIONS: This study presents different techniques to characterize a self-setting calcium phosphate cement and provides experimental data on porosity, mechanical properties and failure. The investigated material possessed an open porosity at its dried state with negligible residual strains and its Young's modulus, obtained from micro-indentation tests, increased with hardening time. The failure loci may be described by a Mohr-Coulomb criterion, characteristic of soil and rock materials.

Characterization of the mechanism of protein glycosylation and the structure of glycoconjugates in tissue culture trypomastigotes and intracellular amastigotes of Trypanosoma cruzi.

Trypomastigote cells of Trypanosoma cruzi incubated with [U-14C]glucose accumulated dolichol-P-P-linked Man7GlcNAc2 and Man9GlcNAc2. Evidence is presented indicating that both oligosaccharides were transferred to asparagine residues in proteins. On the other hand, intracellular amastigotes behaved as epimastigotes, i.e., only Man9GlcNAc2 accumulated and was transferred to proteins under similar incubation conditions. Intracellular amastigotes differed, therefore, from amastigotes obtained from an axenic culture, which behaved as trypomastigotes. A similar processing of protein-linked Man9GlcNAc2 and Man8GlcNAc2 occurred in epimastigotes and trypomastigotes but the structure of the main Man7GlcNAc2 isomer produced by demannosylation of the above mentioned oligosaccharides differed from that of the Man7GlcNAc2 transferred in trypomastigotes and amastigotes from axenic cultures. The infective trypomastigote stage of the parasite showed, therefore, an alteration in the mechanism of protein N-glycosylation when compared to the other stages, namely epimastigote (insect vector stage) and amastigote (mammalian intracellular stage). Complex-type, asparagine-bound oligosaccharides were found to be synthesized in both epimastigotes and trypomastigotes but the amounts of those compounds were extremely low when compared to those of high mannose-type oligosaccharides.

Non-occupational lead exposure and hypertension in northern Italy.

BACKGROUND: Previous research has addressed the issue that low-level blood lead concentration could be associated with an increased risk of hypertension. METHODS: This paper examines the cross-sectional association between blood lead and hypertension in 630 adult males not employed in activities characterized by specific exposure to lead and living in two Northern Italian cities (Verona and Brescia). The participants, aged 26-69 years, constituted a random sample of the patients in the practice of a general practitioner (Verona) and of regular blood donors (Brescia). Logistic regression analysis was used to evaluate the effects of selected variables (blood lead, blood cadmium, zinc erythroprotoporphyrin, haemoglobin, whole blood viscosity, age, body mass index, smoking habits and alcohol consumption) on the probability of being hypertensive. RESULTS: The blood lead concentration (range: 4.3-46.9 micrograms/dl; median: 14.8 micrograms/dl) was very similar in the two samples, whereas the prevalence of hypertension was significantly higher in the Verona sample than in the Brescia sample (20.4% versus 8.3%). Hypertensive subjects showed significantly higher blood lead levels than normotensive ones. In a preliminary (univariate) analysis blood lead levels, body mass index (BMI) and age were each significantly related to the prevalence of hypertension. After adjusting for age and/or BMI, statistical significance of the relationship between blood lead and hypertension was lost.CONCLUSIONS. The present study showed that blood lead is weakly related to hypertension in non-occupationally exposed men. The statistical significance of this association disappeared when age and BMI were used to make adjustments.

Phosphatidylinositol 4-phosphate is associated to extracellular lipoproteic fractions and is detected in tomato apoplastic fluids.

We have recently detected phosphatidylinositol-4-phosphate (PI4P) in the extracellular medium of tomato cell suspensions. Extracellular PI4P was shown to trigger the activation of defence responses induced by the fungal elicitor xylanase. In this study, by applying a differential centrifugation technique, we found that extracellular PI4P is associated with fractions composed of diverse phospholipids and proteins, which were pelleted from the extracellular medium of tomato cell suspensions grown under basal conditions. Using mass spectrometry, we identified the proteins present in these pelleted fractions. Most of these proteins have previously been characterised as having a role in defence responses. Next, we evaluated whether PI4P could also be detected in an entire plant system. For this, apoplastic fluids of tomato plants grown under basal conditions were analysed using a lipid overlay assay. Interestingly, PI4P could be detected in intercellular fluids obtained from tomato leaflets and xylem sap of tomato plants. By employing electrospray ionisation tandem mass spectrometry (ESI-MS/MS), other phospholipids were also found in intercellular fluids of tomato plants. These had a markedly different profile from the phospholipid pattern identified in entire leaflets. Based on these results, the potential role of extracellular phospholipids in plant intercellular communication is discussed.

[Vitamin D intoxication in infants born from Latin-American immigrants. Series of 3 cases]. / Intoxicación por vitamina D en hijos de inmigrantes latinoamericanos. Serie de 3 casos.

Vitamin D intoxication is a well-known cause of hypercalcemia in children and can have serious consequences (renal, cardiac and neurologic mainly). The use of the so-called over-the-counter (OTC) supplements involves a high risk in this taking place. The clinical expression of hypercalcaemia is unspecific, and, together with the fact that the administration of such supplements is frequently denied, the diagnosis of vitamin D intoxication is often delayed and the number of complementary tests performed is high. We here-for expose a series of 3 cases all of which are infants born from Latin-American immigrants who were receiving supplements that came from their parents originary countries. All 3 cases were admitted in our hospital within a period of 5 months. After the first preceding case, the diagnosis of the 2 latter ones was performed promptly and so was the instauration of the treatment for hypercalcemia. The initial levels of serum calcium and of 25-hydroxy vitamin D where, respectively for each case: 17.9 mg/dl and 504 ng/ml; 14.46 mg/dl and 505 ng/ml; 14.2mg/dl and 530 ng/ml. All 3 patients received intravenous treatment with serum, furosemide and corticoids and in one case with subcutaneous calcitonine as well. The clinical outcome was optimal for them all, with normalization of the calcium levels and of the renal function.

Multiscale modeling of composite materials: a roadmap towards virtual testing.

A bottom-up, multiscale modeling approach is presented to carry out high-fidelity virtual mechanical tests of composite materials and structures. The strategy begins with the in situ measurement of the matrix and interface mechanical properties at the nanometer-micrometer range to build up a ladder of the numerical simulations, which take into account the relevant deformation and failure mechanisms at different length scales relevant to individual plies, laminates and components. The main features of each simulation step and the information transferred between length scales are described in detail as well as the current limitations and the areas for further development. Finally, the roadmap for the extension of the current strategy to include functional properties and processing into the simulation scheme is delineated.

The cytotoxic properties of a plant lipid transfer protein involve membrane permeabilization of target cells.

AIMS: To determine whether Ha-AP10, a member of the plant lipid transfer proteins (LTPs) family produces a direct cytotoxic effect on fungal cells mediated by membrane permeabilization. LTPs can inhibit fungal growth and are considered members of the ubiquitous class of antimicrobial peptides. However, the way they exert their effects on target cells is not yet understood. METHODS AND RESULTS: Viability assays demonstrate that Ha-AP10 acts as a fungicidal compound but no harmful effect is observed on plant cells. Liposome leakage assays show that the protein induces a moderate release of fluorescent probes encapsulated in model membranes, indicating its ability to interact with phospholipids. Using a fluorescent indicator of damage at the membrane level, we demonstrate that Ha-AP10 is able to induce the permeabilization of intact fungal spores in a dose-dependent manner. CONCLUSION: The results presented here demonstrate the permeabilization of fungal spores caused by Ha-AP10. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first demonstration of fungal membrane damage by an LTP, giving a clue to elucidate the basis of its antimicrobial properties.

[Probabilistic models to analyze psychiatric contacts]. / Modelli probabilistici per l'analisi dei contatti psichiatrici.

OBJECTIVE: To present three probabilistic models (Poisson, negative binomial, Waring) to analyze the distribution of the number of contacts of patients followed using a psychiatric case register. DESIGN: Longitudinal to obtain the distribution of the number of contacts (during 91 days following that of the first contact) observed on patients followed using the South-Verona Psychiatric Case Register during the period 1/1/79-31/12/91. RESULTS: There were a total number of 6913 contacts on 3454 subjects. The chi-square test for the goodness of fit yields a significant result both for the Poisson distribution (3580 with 6 degrees of freedom, p < 0.001) and for the negative binomial distribution (65.47 with 18 degrees of freedom, p < 0.001); on the other hand a non significant result was obtained for the Waring distribution (25.31 with 19 degrees of freedom, p = 0.151). CONCLUSIONS: The Poisson distribution gave a very poor fit for the distribution of contacts. The negative binomial distribution could be employed to analyze the pattern of contacts when the right tail of the distribution is not important. The Waring distribution is the best of the three presented. Moreover, the variance of the Waring distribution can be decomposed in three components: a random component, a component which accounts for endogenous factors and another component which accounts for esogenous factors. Therefore the Waring distribution is useful when one wants to make comparisons between psychiatric case registers of the same country or of different countries.

Synthesis of dolichol derivatives in trypanosomatids. Characterization of enzymatic patterns.

We have previously described that in certain parasitic protozoa, namely the trypanosomatids, the dolichol-P-P-linked oligosaccharides synthesized in vivo and transferred to protein are devoid of glucose residues and contain 6, 7, or 9 mannose units depending on the species. We have now conducted a cell-free characterization of the enzymatic patterns responsible for these phenotypes. Microsomes from Trypanosoma cruzi, Crithidia fasciculata, Leishmania enriettii, and Blastocrithidia culicis were found to synthesize dolichol-P-[14C]Man but not dolichol-P-[14C]Glc when incubated with rat liver dolichol-P and GDP-[14C]Man or UDP-[14C]Glc, thus providing for an explanation to the absence of glucosylated dolichol-P-P derivatives. Formation of dolichol-P-P-oligosaccharides was assayed in incubation mixtures containing rat liver dolichol-P, GDP-[14C]Man, microsomes, and unlabeled Man5-8GlcNAc2-P-P-dolichol from bovine liver. Membranes from species synthesizing dolichol-P-P-linked Man6GlcNAc2 or Man7GlcNAc2 in vivo were found to synthesize the same compounds but not the higher homologues in the cell-free assay. Species forming Man9GlcNAc2-P-P-dolichol in vivo were found to synthesize lipid-linked Man7GlcNAc2, Man8GlcNAc2, and Man9GlcNAc2 in vitro. It is concluded that there are at least three and probably four different dolichol-P-Man-dependent enzymatic activities involved in the synthesis of dolichol-P-P-linked Man9GlcNAc2 and that microorganisms not forming that compound are devoid of all mannosyltransferases responsible for the addition of the missing residues and not only of the enzyme involved in the synthesis of the homologue higher than the oligosaccharide occurring in vivo by a single mannose unit.

Cytoplasmic male sterility in sunflower: comparison of molecular biology and genetic studies.

The genetics of male fertility restoration and the RFLP of mitochondrial DNA were studied for 16 sunflower cytoplasms (15 male-sterile and a male-fertile). Male fertility restoration/male sterility maintenance patterns distinguished 12 cytotypes. Four cytoplasms were completely unrestored so they were not distinguished genetically. The sunflower lines, tested for their restorer/maintenance reaction, showed that there was a continuous range between 0% and 100% of restorer genotypes according to the CMS considered. Restoration/maintenance patterns indicated that at least some restorer genes are specific to certain CMS. RFLP of mitochondrial DNA revealed specific differences between the cytotypes studied. Three restriction enzymes and 12 probes permitted distinction of 13 cytotypes. No relationship exists between CMS cytotypes and the species from which they originated. For genetical and mitochondrial RFLP studies, phenograms were constructed according to the similarity indexes between cytotypes. Most of the CMS defined by restoration patterns correspond with a restriction fragment pattern of mitochondrial DNA.