Genetic variants identified in novel candidate genes for anorexia nervosa and analysis of molecular pathways for diagnostic applications.

OBJECTIVE: Anorexia nervosa (AN) is a severe psychiatric disorder characterized by an intense fear of gaining weight, a relentless pursuit of thinness, and a distorted body image. Recent research highlights the substantial contribution of genetics to AN's etiology, with genes like BDNF, SLC6A4, and DRD2 implicated. However, a comprehensive genetic test for AN diagnosis is lacking. This study aims to elucidate the biological foundations of AN, examining variants in genes associated with syndromic forms, rare variants in AN patients, and candidate genes from GWAS studies, murine models, or established molecular pathways. MATERIALS AND METHODS: The study involved 135 AN patients from Italy, diagnosed based on DSM-V criteria. A specialized Next-Generation Sequencing panel targeting 163 genes was designed. Sequencing was performed on an Illumina MiSeq System, and variants were analyzed using bioinformatics tools. Data on clinical parameters, exercise habits, and AN types were collected. RESULTS: The AN cohort, predominantly female, exhibited diverse clinical characteristics. Our analysis identified gene variants associated with syndromic forms of AN, such as STRA6, NF1, MAT1A, and ABCC6. Variants were also found in known AN-related genes (CD36, DRD4, GCKR, GHRL, GRIN3B, GPR55, LEPR) and in other 16 candidate genes (A2M, AEBP1, ABHD4, ACBD7, CNTNAP, GFRAL, GRIN2D, LIPE, LMNA, NMU, PDE3B, POMC, RYR1, TNXB, TYK2, VPS13B), highlighting the complexity of AN's genetic landscape. The endocannabinoid and dopamine pathways play crucial roles. Skeletal muscle-related genes and appetite-regulating hormones also revealed potential connections. Adipogenesis-related genes suggest AN's association with subcutaneous adipose tissue deficiency. CONCLUSIONS: This study provides comprehensive insights into the genetic underpinnings of AN, emphasizing the importance of multiple pathways. The identified variants contribute.

Proinflammatory circulating markers: new players for evaluating asymptomatic acute cardiovascular toxicity in breast cancer treatment.

PURPOSE: This study aimed to evaluate markers of cardiac damage (total CK, CKMB and CRP), inflammatory markers (free iron, homocysteine and TNF-α) as well as lipidogram in breast cancer patients undergoing acute cycles of doxorubicin (DOX), paclitaxel (PTX) or trastuzumab (TZ) and to verify if there is an association between these markers and the toxicity of the chemotherapeutic treatment. Methods: Included in the study were 120 breast cancer patients and 50 healthy controls. All analyzes were performed on automated systems. For the statistical analysis, each group was compared with the controls according to their normality by Student's t-test and Mann-Whitney test. Results: Our results showed that DOX treatment led to increased hsCRP (4.80 ± 1.23 mg/dL, p = 0.0005), triglycerides (187.6 ± 25.06, p = 0.0231), TNF-α (42.31 ± 17.96 pg/mL, p = 0.01) and Fe levels (138.8 ± 18.6 µg/dL, p = 0.0193). In the meantime, PTX induced changes in CK-MB (8.78 ± 4.2 U/L, p = 0.0361), hsCRP (7.12 ± 1.87 mg/dL, p = 0.0006), cholesterol (201.7 ± 19.54, p = 0.05), triglycerides (201.7 ± 19.54, p = 0.0277), TNF-α (38.27 ± 9.12 pg/mL, p = 0.023), homocysteine (10.95 ± 0, 86 µmol/L, p = 0.005), and free iron (113 ± 18 6 µg/dL, p = 0.045) while TZ augmented CK-MB (6.9 ± 1.97 U/L, p < 0.00), hsPCR (3.12 ± 0.68 mg/dL, p = 0.095), cholesterol (218.3 ± 16.79, p = 0.0317), triglycerides (218.3 ± 16.79, p = 0.0127), TNF-α (89.6 ± 12.11, p = 0.032), homocysteine (9.95 ± 1.15 µmol/L, p = 0.0396), free iron (120.5 ± 4.64 µg/dl, p = 0.0058) as well. Conclusions: Our data demonstrated the existence of a proinflammatory net triggered by breast cancer chemotherapy that could increase cardiomyocytes permeability and allow the leakage of circulating proteins as CK-MB and induce the production of hsCRP.

Cathodoluminescent screen imaging system for seeded blob detection in toroidal plasma experiment.

We designed and built a diagnostic based on a cathodoluminescent screen for the detection of turbulent plasma structures with high spatial resolution. The screen is coated with a low threshold energy cathodoluminescent powder that emits light when exposed to a plasma. The emitted light is imaged with a fast frame camera combined with an image intensifier and an optical bandpass filter. The diagnostic is used to study turbulent structures and seeded blobs. The results are analyzed with pattern recognition algorithms to track the turbulent structures and study their evolution in time.

Optically isolated millimeter-wave detector for the Toroidal Plasma Experiment.

We have designed and built an optically isolated millimeter-wave detection system to prevent interference from a nearby, powerful, 2.45 GHz microwave source in millimeter-wave propagation experiments in the TORoroidal Plasma EXperiment (TORPEX). A series of tests demonstrates excellent system noise immunity and the ability to observe effects that cannot be resolved in a setup using a bare Schottky diode detector.

The students' reasons to choose a nursing degree program: an Italian exploratory study.

BACKGROUND: From the international literature very interesting cues emerge about students' motivations to choose a Nursing Degree Program. Unfortunately, this phenomenon is largely unexplored in Italy. An observational study was conducted at the University of Pavia, with the aim to investigate the reasons underpinning the students' choice to enter a Nursing Degree Program. METHODS: A semi-structured, self-administrated questionnaire was used for this single-center cross-sectional study. The questionnaire was aimed to investigate the reasons underpinning the students' choice to enter a Nursing Degree Program, using both open-ended and close-ended multiple choice questions. Descriptive statistics have been used to describe collected data. Open-ended response analysis was conducted through an exploratory and qualitative analysis of language. RESULTS: Response rate was 71% (196/275). Our study results highlighted a variety of reasons that encourage students to begin a Nursing Degree Program: the feeling of usefulness (80%), the desire to help and to care people (78%), the interest in the sciences (71%), the compassion to the suffering people (66%). We also identified 4 categories that describe which characteristics a nurse should have according to the students' point of view: expertise; personal characteristics; to experience the professional life as a social function and to have interest in the health field. CONCLUSIONS: Students' answers indicate that often the choice to enter a Nursing Degree Program is not supported by clear ideas and strong motivations. We consider it crucial to provide a realistic image of the nursing role and opportunities for career development, so that students can have the right elements to make a conscious choice. There is a need for more qualitative research to explore the reasons why students choose the Nursing Degree Program; moreover, to identify, from the beginning of the course, those students who are in crisis of motivation, in order to adopt support strategies that could enable them to successfully achieve academic career.

Acute exposure to essential amino acids (EAA) activates MTOR/p70 signaling in soleus muscle of chronically EAA-treated aged rats.

Using an in vitro assay we assessed whether the acute exposure of soleus muscle of adult and aged rats to essential amino acid enriched mixture (EAAem) activates mTOR signaling pathway (mTOR and p70S6K) even after prolonged supplementation with the same mixture. A total of 20 adult (9 months of age at the end of treatment) and 20 aged (18 months of age at the end of treatment) male Wistar rats were used. Ten of each group were treated with EAAem (1.5 gr/kg/day in tap water) for 6 months. At the end of treatment the rats were grouped (n = 5 each group) as follows: adult (AD) and aged (AG) untreated controls; adult (AD_EAAem) and aged (AG_EAAem) chronically supplemented with EAAem; adult (AD+EAAem) and aged (AG+EAAem) acutely incubated with EAAem (soleus in 1 percent EAAem for 30 min); AD_EAAem+ and AG_EAAem+ acutely incubated with EAAem. Following treatment the activation level of mTOR and p70S6K was measured by Western blot. The basal level of mTOR and p70S6K activation appeared to be higher in AD compared with AG. In AG+EAAem a significant change in the level of p70S6K activation, unlike mTOR, was observed whereas no change was observed in AD+EAAem. In AD_EAAem muscles the basal level of p70S6K activation, unlike mTOR, was significantly lower than in AD and the acute exposure to EAAem produced a significant reduction of mTOR activation. Contrarily to AG, in AG_EAAem+ the acute exposure to EAAem produced a significant activation of mTOR, unlike p70S6K. Results in the adults indicated a higher basal level of activation and a lower responsiveness of the pathway to acute and chronic exposure to EAA-enriched mixture. On the contrary, in the aged, a lower basal level of activation was associated with a higher responsiveness to EAAem. In particular, although with a different timing, acute exposure to EAAem activated mTOR signaling even following prolonged supplementation.

Origin of a polyploid accession of Brachiaria humidicola (Poaceae: Panicoideae: Paniceae).

Brachiaria humidicola, a species adapted to poorly drained and infertile acid soils, is widely used throughout the tropics. Cytological characterization of 54 accessions of B. humidicola for breeding purposes revealed 2n = 36, 42, and 54 chromosomes. One accession (H030), with 2n = 42 chromosomes, showed a different meiotic behavior. In most accessions from the genus Brachiaria previously studied, the basic chromosome number is x = 9, but the putative basic number in H030 appears to be x = 6. Since six univalent chromosomes were found in diakinesis and metaphase I, and these behaved as laggards in anaphase I, it was hypothesized that both genitors were derived from x = 6, and that this accession is a heptaploid 2n = 7x = 42. The parental genomes did not have the same meiotic behavior, particularly during anaphase, when one genome consisting of six univalents remained as laggards and underwent sister-chromatid segregation. At telophase, 18 segregated chromosomes were found at each pole. The laggard genome did not reach the poles at telophase I or II in time to be included in the nucleus and was eliminated as micronuclei.

Condylar-mandibular asymmetry, a reality.

The aim of this study is to evaluate the possibility to recognize a condylar-mandibular asymmetry through a panoramic radiograph. Results from a previous work, in which 100 skulls from the Museum of the Institute of Anatomy of the University of Pavia were studied and measured, showed the presence of asymmetry. Using the same skulls we examined the possible correlation between morphological and radiological data. We did not find out correlation's between the condylar asymmetry evaluated at the anatomical level and the radiological asymmetry which was indeed found. This is probably due to the different positioning of the jaws during the two different measuring processes. Nevertheless our results confirm the daily experience of dentistry: asymmetry of mandibular condyle can be one of the mayor causes for the asymmetry of the stomatognatic apparatus.

[Prevalence of obesity amount elementary school children in Grenoble]. / Prévalence de l'obésité chez les enfants scolarisés en école primaire à Grenoble.

Facing the growing increase of childhood obesity in France, the Grenoble Municipal School Health Service wanted to evaluate the importance of this phenomenon present among elementary school children in the area. A descriptive, cross-sectional and retrospective survey was conducted during the 2000/2001 academic year among a group of 1361 children born in 1990. The Quetelet Index was utilized to measure the level of being overweight, and the breaking point levels for being overweight and obese were defined according to new international standards. The prevalence of obesity is approximately 3% (2.7% in girls and 2.9% in boys) and remains constant from nursery school to elementary school. Nearly 35% of obese children in nursery school remain obese at the end of elementary school. Overweight prevalence is at its highest and increases significantly from nursery school (8.3%) through elementary school (14%). The study further revealed that girls aged 10-12 who did not pass and were held back from the next grade are five times more at risk of obesity than those who pass. The Municipal School Health Service has a critical role to play in the prevention of childhood obesity and its early screening and detection.

Repeated epidemics caused by extended-spectrum beta-lactamase-producing Serratia marcescens strains.

An outbreak of Serratia marcescens involving 42 patients admitted to the general intensive care unit of the Hospital of Varese, Italy, occurred from March 1994 to August 1995. The causative strains were resistant to oxyimino-cephalosporins and monobactams due to their production of an extended-spectrum beta-lactamase. Another outbreak caused by Serratia marcescens strains had occurred in the same unit a few months earlier, from February to October 1993, with the strains involved producing a novel TEM-derived extended-spectrum beta-lactamase. In order to verify whether there were any relationships between isolates from the two epidemics, the strains and their enzymes were characterized. Biochemical data and gene amplification experiments showed that the isolates of the second outbreak harbored a non-conjugative plasmid of approximately 48 kb, codifying for the production of an SHV-derived extended-spectrum beta-lactamase with pI 8.2. Restriction fragment length polymorphism analysis of total genomic DNA by pulsed-field gel electrophoresis of Serratia marcescens isolates unambiguously identified two different bacterial clones responsible for the two epidemics. Epidemiological and microbiological investigations demonstrated the long persistence of Serratia marcescens strains and their circulation in other hospital wards, thus suggesting their possible role as a long-term reservoir for further epidemic spread.

Comparative activity of piperacillin/tazobactam against clinical isolates of extended-spectrum beta-lactamase-producing Enterobacteriaceae.

beta-Lactam resistance on the part of the Enterobacteriaceae causes serious therapeutic problems in our institutions due to their production of extended-spectrum beta-lactamases (ESbetaLs). We studied the in vitro activity of beta-lactam/beta-lactamase inhibitor combinations and third-generation cephalosporins and monobactams against 71 clinically relevant Enterobacteriaceae which produced TEM- and SHV-derivative ESbetaLs. Of the single drugs and combinations tested, piperacillin/tazobactam proved to be the most effective. Piperacillin/tazobactam was highly active against Proteus mirabilis, with minimum inhibitory concentrations (MICs) ranging from 0.125 to 16 microg/ml; Escherichia coli (MICs from 2 to 16 microg/ml) and Serratia marcescens (MICs from 4 to 8 microg/ml), while its activity against Klebsiella pneumoniae ESbetaL producers turned out to be closely related to the type and the amount of enzyme produced, the MIC ranging from 1 to 128 microg/ml. The antibacterial activity of piperacillin/tazobactam was stronger than that of ticarcillin/clavulanate, ceftriaxone, cefotaxime, ceftazidime and aztreonam, and the combination shared favorable in vitro activity properties against the ESbetaL producers with imipenem which, however, should be kept as reserve product.

Outbreak of extended-spectrum beta-lactamase producing Serratia marcescens in an intensive care unit.

Serratia marcescens has recently been identified as an important etiological agent in nosocomial infections, and is considered to be an opportunistic pathogen agent in immunosuppressed patients undergoing long periods of intensive care. Research carried out in 1991 and 1992 showed that it was of epidemiological relevance in only 1-2% of clinical isolates at the Ospedale di Circolo, Varese, Italy. However, between 7 February and 11 October 1993, the incidence of cases attributable to S. marcescens had increased to 5%; 157 strains of Serratia marcescens were isolated from clinical specimens of 43 patients admitted to an intensive care unit; these strains, characterized by epidemic spread, showed the same pattern of multiresistance to antibiotics including monobactams and oxyimino-cephalosporins. During the same period 23 isolates were also recovered from 18 patients admitted to wards other than the intensive care unit; these strains, characterized by a wide range of antibiotic susceptibility, were also sensitive to beta-lactam antibiotics with the exception of first generation cephalosporins. The production of extended-spectrum beta-lactamases (ES beta Ls) and their genetic determinism were studied. All the epidemic strains of S. marcescens resistant to ceftazidime, cefotaxime, ceftriaxone and aztreonam produced three different beta-lactamases with pI 5.4, 5.5 and 8.4 respectively. In contrast, non-epidemic strains produced only a beta-lactamase with pI 8.4. The beta-lactamase with pI 5.5 was plasmid-mediated, hydrolizing ceftazidime and aztreonam, showing it to be an ES beta L; while the beta-lactamase with pI 5.4, although plasmid-mediated, did not hydrolize monobactams or oxyimino-cephalosporins.(ABSTRACT TRUNCATED AT 250 WORDS)

The UL8 component of the herpes simplex virus helicase-primase complex stimulates primer synthesis by a subassembly of the UL5 and UL52 components.

The herpes simplex virus type 1 (HSV) UL5, UL8, and UL52 proteins form a helicase-primase complex in infected cells. Several laboratories have demonstrated that helicase and nucleoside triphosphatase activities of the heterotrimer (UL5/8/52) are indistinguishable from that of a subassembly of UL5 and UL52 (UL5/52). Although the UL5/52 subassembly functions in coupled primase-polymerase assays on homopolymeric templates, its activity on natural DNA templates has been reported to require UL8. To determine the role of UL8 in primase assays, the activity of the UL5/52 subassembly was compared to that of the heterotrimer reconstituted by adding UL8 to UL5/52. We detected significant activity of the UL5/52 subassembly in coupled primase-polymerase and oligoribonucleotide primer synthesis assays on phi X174 and M13 virion DNAs. However the addition of UL8 to UL5/52 stimulated this activity in a dose-dependent manner. We demonstrate that stimulation occurred at the level of primer synthesis. UL8 did not affect the amount or size of primers annealed to template, their utilization by DNA polymerase, or the use of specific initiation sites within the template. In kinetic studies, the rate of primer synthesis was increased by UL8 but the Km for phi X174 DNA template was unchanged. These results suggest that a function of the UL8 component of the HSV helicase-primase complex is to increase the efficiency of primer synthesis by UL5/52.

Deletions of the carboxy terminus of herpes simplex virus type 1 UL42 define a conserved amino-terminal functional domain.

The herpes simplex virus type 1 UL42 protein was synthesized in reticulocyte lysates and assayed for activity in vitro. Three functional assays were used to examine the properties of in vitro-synthesized UL42: (i) coimmunoprecipitation to detect stable complex formation with purified herpes simplex virus type 1 DNA polymerase (Pol), (ii) a simple gel-based assay for DNA binding, and (iii) a sensitive assay for the stimulation of Pol activity. UL42 synthesized in reticulocyte lysates formed a stable coimmunoprecipitable complex with Pol, bound to double-stranded DNA, and stimulated the activity of Pol in vitro. Carboxy-terminal truncations of the UL42 protein were synthesized from restriction enzyme-digested UL42 gene templates and gene templates made by polymerase chain reaction and assayed for in vitro activity. Truncations of the 488-amino-acid (aa) UL42 protein to aa 315 did not abolish its ability to bind to Pol and DNA or to stimulate Pol activity. Proteins terminating at aas 314 and 313 showed reduced levels of binding to Pol, but these and shorter proteins were unable to bind to DNA or to stimulate Pol activity. These results suggest that all three of the biochemical functions of UL42 colocalize entirely within the N-terminal 315 aas of the UL42 protein. Amino acid sequence alignment of alpha herpesvirus UL42 homologs revealed that the N-terminal functional domain corresponds to the most highly conserved region of the protein, while the dispensable C terminus is not conserved. Conservative aa changes at the C terminus of the 315-aa truncated protein were used to show that conserved residues were important for activity. These results suggest that 173 aa of UL42 can be deleted without a loss of activity and that DNA-binding and Pol-binding activities are correlated with the ability of UL42 to stimulate Pol activity.

Mutations in the C terminus of herpes simplex virus type 1 DNA polymerase can affect binding and stimulation by its accessory protein UL42 without affecting basal polymerase activity.

We have analyzed the effects of mutations in the herpes simplex virus type 1 DNA polymerase (Pol) C-terminal UL42 binding domain on the activity of Pol and its ability to form complexes with and be stimulated by UL42 in vitro. Wild-type Pol expressed in Saccharomyces cerevisiae was both bound and stimulated by UL42 in vitro. C-terminal truncations of 19 and 40 amino acids (aa) did not affect the ability of Pol to be stimulated by UL42 in vitro. This stimulation as well as basal Pol activity in the presence of UL42 was inhibited by polyclonal anti-UL42 antiserum, thus indicating a physical interaction between Pol and UL42. Removal of the C-terminal 59 aa of Pol and internal deletions of 72 aa within the Pol C terminus eliminated stimulation by UL42. None of the truncations or deletions within Pol affected basal polymerase activity. In contrast with their ability to be stimulated by UL42, only wild-type Pol and Pol lacking the C-terminal 19 aa bound UL42 in a coimmunoprecipitation assay. These results demonstrate that a functional UL42 binding domain of Pol is separable from sequences necessary for basal polymerase activity and that the C-terminal 40 aa of Pol appear to contain a region which modulates the stability of the Pol-UL42 interaction.

Isolation of a coenzyme M-auxotrophic mutant and transformation by electroporation in Methanococcus voltae.

An auxotrophic mutant of Methanococcus voltae was isolated that required coenzyme M (CoM) for growth. With the mutant as a recipient, conditions were developed that allowed the introduction of wild-type CoM+ DNA into the mutant methanogen via electroporation. This method also allowed the rescue of both a histidine and purine auxotroph as well as the introduction of DNA determining resistance to the CoM analog 2-bromoethanesulfonic acid. Electroporation of the CoM(+)-determining DNA was 50- to 80-fold more efficient than natural transformation.

[Cardiac effects of exhausting isometric muscular contraction in trained and endurance athletes]. / Effetti cardiaci della contrazione muscolare isometrica ad esaurimento in atleti di potenza e di resistenza.

The effect of exhausting isometric contraction (60% of the maximal voluntary contraction) on left ventricular function has been investigated using echocardiography (M and B mode) in 4 groups, each of 8 subjects (27 +/- 5 years; mean +/- SD): weight lifters, 2 years of training; amateur cyclists, 4 years of training; sedentary controls, and hypertensive subjects, 1st WHO class. Heart rate, arterial pressure, ventricular diameters (diastolic and systolic), wall thickness, wall stress and the double product (heart rate x arterial pressure) were determined at rest, at exhaustion and after 30, 90 and 180 s of recovery. Maximal voluntary contraction was 48 +/- 9 kg in weight lifters; 32 +/- 5 kg in amateur cyclists; 32 +/- 4 kg in sedentary control and 36 +/- 7 kg in 1st WHO class. Exhaustion time ranged from 80 s to 120 s. Our results showed that: 1) in all subjects at exhaustion, arterial pressure and heart rate increased significantly and the index of ventricular function was decreased but not significantly; 2) after 180 s of recovery both arterial pressure and heart rate returned to the pre-exercise value; 3) wall stress was significantly lower in weight lifters than in sedentary controls; 4) the double product (index of myocardiac oxygen consumption) resulted lower in amateur cyclists that in sedentary controls. The hypertrophy of the heart could justify the slight increase in wall stress observed in weight lifters during isometric exercise. However, the low index of myocardiac oxygen consumption in amateur cyclists probably reflects the changes in cardiovascular function due to the endurance training. In hypertensive subjects, the isometric contraction causes significant and substantial increase of the wall stress and the double product.