Suitability of miRNA assessment in postmortem interval estimation.

OBJECTIVE: The aim of this review was to explore recent pieces of evidence focused on the use of miRNAs for PMI estimation both in humans and animal experiments, with particular interest on the best miRNAs to use as reference/target markers in different tissues or biological fluids. MiRNAs are innovative biomarkers used in clinical and research field; they appear very attractive, being introduced in forensic research scenarios even for PMI estimation. MATERIALS AND METHODS: Data from PubMed and Scopus were analyzed from January 2013 to August 2020. Based on inclusion/exclusion criteria, high-quality articles have been selected to become the subject of this review. RESULTS: A total of 737 papers were found but, after titles/abstracts screening for inclusion criteria and a full-text careful selection, 33 papers were deeply studied. After the exclusion of 19 papers, 15 articles remained. Eight papers dealt with animals (mice/rats), two both with animals and humans (for method validation previously built), while 5 exclusively with humans. Myocardium (6/15) and brain (6/15) were the most studied tissues, respectively in mice/rats and humans. PMI considered was up to 7.5 days in mouse studies and less than 3 days in human models. CONCLUSIONS: Because of their significant stability in both early and long PMI, miRNAs are the cleverest reference markers to be used. Temperature and environmental conditions influence mostly mRNA, while miRNAs are less susceptible to them. The best miRNA to choose depends on its tissue specificity, i.e., miR-9 and miR-125 in brain or miR-1 and miR-133 in skeletal muscle/heart.

Use of cognitive enhancers: methylphenidate and analogs.

OBJECTIVE: In the last decades, several cognitive-enhancing drugs have been sold onto the drug market. Methylphenidate and analogs represent a sub-class of these new psychoactive substances (NPS). We aimed to review the use and misuse of methylphenidate and analogs, and the risk associated. Moreover, we exhaustively reviewed the scientific data on the most recent methylphenidate analogs (methylphenidate and ethylphenidate excluded). MATERIALS AND METHODS: Literature search was performed on methylphenidate and analogs, using specialized search engines accessing scientific databases. Additional reports were retrieved from international agencies, institutional websites, and drug user forums. RESULTS: Methylphenidate/Ritalin has been used for decades to treat attention deficit disorders and narcolepsy. More recently, it has been used as a cognitive enhancer and a recreational drug. Acute intoxications and fatalities involving methylphenidate were reported. Methylphenidate was scheduled as an illegal drug in many countries, but NPS circumventing the ban and mimicking the psychostimulant effects of methylphenidate started being available: ethylphenidate, 3,4-dichloromethylphenidate, 3,4-dichloroethylphenidate, 4-fluoromethylphenidate, 4-fluoroethylphenidate, methylnaphthidate, ethylnaphthidate, isopropylphenidate, propylphenidate, 4-methylmethylphenidate, and N-benzylethylphenidate have been available in the past few years. Only little data is currently available for these substances. Many intoxications involving methylphenidate analogs were reported. To date, ethylphenidate was involved in 28 fatalities, although it was reportedly directly related to the cause of death in only 7 cases; 3,4-dichloroethylphenidate was involved in 1 death. CONCLUSIONS: The rapid expansion of methylphenidate analogs onto the drug market in the past few years makes likely the occurrence of intoxications and fatalities in the next years. Careful monitoring and systematic control of methylphenidate analogs should be undertaken to reduce the uprising threat, and education efforts should be made among high-risk populations.

Synthetic cathinones related fatalities: an update.

OBJECTIVE: Synthetic cathinones, more commonly known as "bath salts", are synthetic drugs chemically related to cathinone, a psychostimulant found in the khat plant. They are the first most consumed products among new psychoactive substances, which cause psychostimulant and hallucinogenic effects determining a number of fatalities worldwide.  In this paper, we have systematically reviewed cases of synthetic cathinones-related fatalities analytically confirmed, which have occurred in the last few years. MATERIALS AND METHODS: Relevant scientific articles were identified in Medline, Cochrane Central, Scopus, Web of Science and Institutional/government websites up to November 2017 using the following keywords: synthetic cathinones, mephedrone, methylenedioxypyrovalerone, MDPV, methylone, ethylone, buthylone, fatal intoxication, fatalities and death. RESULTS: In total, 20 citations met the criteria for inclusion, representing several fatal cases with analytically confirmed synthetic cathinones in biological sample/s of the deceased. The death was attributed to hyperthermia, hypertension, cardiac arrest and more in general to the classic serotonin syndrome. Only rarely did the concentration of the parent drug causing fatality overcome the value of 1 mg/L in post-mortem biological fluids. CONCLUSIONS: Abuse of synthetic cathinones still represents a serious public health issue. Systematic clinical studies on both the animal and human model are lacking; therefore, the only available data are from the users who experience the possible hazardous consequences. Analytical methodologies for the identification of parent compounds and eventual metabolites both in ante-mortem and post-mortem cases need to be developed and validated. Analytical data should be shared through different communication platforms with the aim of stopping this serious health threat for drug users.

Flutamide-induced hepatotoxicity: ethical and scientific issues.

OBJECTIVE: Flutamide (FLU) is a non-steroidal antiandrogen drug approved for the treatment of advanced prostate cancer. While this indication limits the use to male patients, FLU is widely prescribed to women, off-label, for the treatment of polycystic ovary syndrome (POCS) related hirsutism and acne. According to the literature, its assumption is associated with a higher incidence of adverse events in women than in male patients. MATERIALS AND METHODS: A literature search was conducted in main databases targeting unwilling FLU effects in hepatic and reproductive function. References in the selected paper were also considered as an additional source of data. Human- and animal-based studies were separately considered. RESULTS: Twenty-three human-based studies were evaluated: ten were case reports, six were retrospective studies, four were prospective, two were surveillance studies, while the last was an observational study. Nine animal-based studies were also evaluated. CONCLUSIONS: Scientific contributions highlight that FLU is responsible for specific hepatotoxic profiles in the female gender. From the ethical point of view, off-label prescribing of FLU in women is not only substantially unlawful, but also, without major safeguards being granted, a potential source of liability for prescribers.

"Vehicular homicide", a new Italian offence: law provisions and comparison with some other European countries.

According to the most recent World Health Organization report, road accidents represent a very serious public health issue, claiming each year more than 1.2 million lives worldwide and being the leading cause of death among young people aged between 15 and 29 years. Up to now, the policies to reduce this issue are different, unbalanced and often inadequate not only in Italy, but also in the other European Countries. Specifically, the Italian Parliament has recently introduced a new law (n. 41 of March 23rd, 2016), making vehicular homicide together with road traffic injuries a criminal offense, both to be punished as a result of negligence. The measure came into force on March 25th, 2016. In this paper, the provisions of the above-reported law have been assessed, taking into account the modifications introduced in the Penal Code by this law and the impact it is having and will have on drivers on a day-to-day basis. Similarities and differences with legislative framework of some other European countries were also examined. Finally, some open questions to be solved are named as an open eye for future considerations.

Effects of ketamine on psychomotor, sensory and cognitive functions relevant for driving ability.

Ketamine is a dissociative anesthetic. The misuse of ketamine as a recreational drug has increased over the last decade, especially in rave parties or clubs. Short-term ketamine pilot protocols have been undertaken for treatment-resistant depressive clients. In this study, we review and comment on the evidence relating to the potential of ketamine as a causative/contributory factor in traffic accidents. To determine the causal role of ketamine in traffic accidents, a literature search on the psychomotor, cognitive, visual and perceptual functions related to safe driving was conducted. Furthermore, to interpret related data better, an overview of ketamine and its congeners' clinical pharmacology issues, recreational psychoactive effects, and identification in biological specimens is also provided.

Variable phenotypic spectrum of diabetes mellitus in a family carrying a novel KCNJ11 gene mutation.

AIMS: Heterozygous activating mutations in KCNJ11, which encodes the Kir6.2 subunit of the pancreatic ATP-sensitive potassium (K(ATP)) channel, cause both permanent and transient neonatal diabetes. Identification of KCNJ11 mutations has important therapeutic implications, as many patients can replace insulin injections with sulphonylurea tablets. The aim was to determine if a KCNJ11 mutation was responsible for a dominantly inherited form of diabetes mellitus, showing variability in age at diagnosis, in an Italian family. METHODS: We sequenced KCNJ11 in members of a three-generation family with variable phenotypes of dominantly inherited diabetes mellitus. One had transient early-onset diabetes, one had impaired glucose tolerance during the second pregnancy, and two had young-onset diabetes. None of the subjects showed permanent neonatal diabetes or neurological symptoms. RESULTS: A novel heterozygous mutation (c. 679C-->G and c. 680A-->T) was identified, resulting in a GAG-->CTG (E227L) substitution in KCNJ11. Functional studies of recombinant heterozygous K(ATP) channels revealed a small reduction in channel inhibition by ATP (IC(50) of 15 micromol/l and 38 micromol/l for wild-type and heterozygous channels, respectively) and an increase in the resting K(ATP) current. This would be expected to impair insulin secretion. The results are in agreement with the mild phenotype of the patients. CONCLUSIONS: Our results broaden the spectrum of diabetes phenotypes resulting from KCNJ11 mutations. They indicate testing for KCNJ11 mutations should be considered not only for neonatal diabetes but also for other forms of dominantly inherited diabetes with later onset, especially where these are associated with a low body mass index and low birth weight.

Distal movement of maxillary canines and premolars with sectional mechanics following Distal Jet application to molars.

OBJECTIVES: to assess the effectiveness of Distal Jet without simultaneous fixed appliance treatment. SUBJECTS AND METHODS: twenty subjects with Class II malocclusion who received the Distal Jet appliance to move maxillary molars distally. Subsequently, the canine and premolar distal movements were achieved by the use of.017 x.025 Ni-Ti sectional wires and memory power chain. RESULTS: revealed that Distal Jet is more effective with less anchorage loss when this appliance is used independent of multibracket appliances. CONCLUSION: this study showed that Distal Jet is an effective and predictable appliance. The distal movements of upper premolars, canines and incisors with sectional wires fabricated from.017 x.025 Ni-Ti were biomechanically very effective, simple in clinical application, and did not require any patient compliance.

Evaluation of biocompatibility of metallic dental materials in cell culture model.

Biocompatibility of metals for dental use was tested using a three-dimensional model consisting of oral keratinocytes cultured on de-epidermised sub-mucosa. The toxicity of orthodontic metallic wire and soldering material was assessed through parameters such as the morphology and growth rate of the keratinocytes, as well as by classical histology, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The sharp composition of metallic wires and soldering materials was assessed by Auger Electron Spectroscopy (AES). The results of our experiment showed that the new model revealed inhibition of keratinocyte growth and stratification near soldering material, whereas mucosal cells were able to grow and layer out on dental wire. It is concluded that this experimental model, which simulates the oral environment, is useful for studying the effects of materials for dental use for its sensibility and reproducibility. Moreover it can provide morpho-functional information which cannot be achieved by traditional methods.

A functional approach to the TMJ disorders.

This manuscript describes our conservative approach to treatment of TMJ disorders. The method we use had been suggested by Rocabado - its aims are: joint distraction by the elimination of compression, restoration of physiologic articular rest, mobilization of the soft tissues, and whenever possible, to improve the condyle-disk-glenoid fossa relationship. To support these claims two clinical cases are presented where the non-invasive therapy was successful. The results obtained confirm the validity of this functional approach.

[Stickler syndrome: maxillo-facial abnormalities]. / La sindrome di Stickler: anomalie maxillo-facciali.

A case of Stickler syndrome (hereditary arthro-ophthalmo-dystrophy) with maxillo-facial abnormalities and colon atresia is reported. The authors emphasize that in Stickler syndrome, (a chondrodystrophy with congenital alteration of type II collagen) the maxillofacial abnormalities are consecutive to prosencephalic neural crests dysneurulation, which caused mesethmoidal dysneurulation (and consequently of frontonasal-premaxillary structures); eye and colon abnormalities are consecutive to rhombo-mesencephalic neural crests dysneurulation. The Authors hypothesize that genic mutation, responsible of enzymatic deficiency of the neural crest multipotent cells, caused a morphogenetic alteration of the fronto-naso-philtral structures of the midface and of low face structures.

Cranioectodermal dysplasia: a new patient with an inapparent, subtle phenotype.

Cranioectodermal dysplasia is a rare syndrome characterized by craniofacial and skeletal anomalies and ectodermal dysplasia. Life-threatening associated conditions (i.e., kidney failure and abnormal regulation of the parathyroid-bone axis) can also develop. We report a patient whose features are suggestive of an inapparent, subtle phenotype of the syndrome. The patient is a 4-year-old girl with only dolichocephaly and clinodactyly; microdontia, hypodontia, and taurodontia (i.e., cone-shaped teeth); anteverted nares, full cheeks, and everted lower lip; epicanthal folds, hypertelorism and hyperopia; and corpus callosum hypoplasia. She has no rhizomelic limb shortening or hair abnormalities. In view of the rarity of the cranioectodermal dysplasias, the variability of the phenotype, and the uncertain outcome of some previously described patients, we believe this inapparent, subtle case should reported to enable better understanding and treatment of this rare syndrome.

[Alcohol, drugs, pharmacologic agents, and street safety]. / Alcol, droga, farmaci e sicurezza stradale.

Deaths due to road accidents during weekends have become a worrying phenomenon in Italy. With the aim of highlighting the role of psychotropic substances (alcohol, drugs of abuse) in causing road accidents, a survey based on clinical and chemico-toxicological analyses has been carried out on car drivers in the Veneto region during night weekends since 1994. Rapid clinical screening was carried out on 7952 drivers. 1399 of these, suspected to be under the influence of psychotropic substances, were subjected to complete clinical and toxicological ascertainment involving the following procedures: a) anamnesis, aiming at evidence of possible current or past use of psychotropic substances; b) objective clinical examination, aiming at finding evidence of recent (signs of acute or chronic intoxication) or past use (signs of withdrawal or associated organic pathologies) of psychotropic substances; c) double sampling of blood and urine and chemico-toxicological analysis using immunochemical, GC-HS and GC/MS-SIM techniques. As well as many data of social and behavioural interest, processing of results demonstrated that: a) 56.7% of the drivers examined had consumed alcoholic beverages; b) 30.4% had BACs higher than the threshold permitted in Italy (80 mg/100 ml); c) 15.7% of drivers were found to be under the influence of drugs of abuse or psychoactive drugs; d) the most frequently found substances were (in order): cannabinoids, stimulants (cocaine, amphetamines), opiates.

Mucogingival interceptive surgery of buccally-erupted premolars in patients scheduled for orthodontic treatment. II. Surgically treated versus nonsurgically treated cases.

BACKGROUND: The aim of this 2-year longitudinal study was to compare the width of keratinized gingiva after orthodontic therapy for buccally erupting premolars that had been pretreated by extraction of deciduous teeth alone versus interceptive mucogingival surgery. METHODS: In 8 patients (aged 9 to 12 years) who presented with bilateral buccal eruption of homologous teeth (premolars), one side was randomly treated with extraction of the deciduous molar and mucogingival surgery (test site), while the other side was treated only by extraction of the deciduous molar (control site). All of the subjects underwent orthodontic treatment with fixed appliances. RESULTS: At the baseline visit prior to any treatment, there was no significant difference between the mean amount of keratinized gingiva at test sites (3.06 mm) and control sites (2.93 mm). Two years later, upon completion of orthodontic treatment, there was a significant difference between test (2.93 mm) and control (1.37 mm) sites in the mean width of keratinized tissue. In the control (untreated) group, 2 sites exhibited 1 mm of gingival recession after orthodontic treatment. CONCLUSIONS: Mucogingival interceptive surgery is an effective technique to maintain keratinized tissue in correspondence with buccally-erupted teeth.

Effects of single dose of gamma-hydroxybutyric acid and lorazepam on psychomotor performance and subjective feelings in healthy volunteers.

OBJECTIVE: This study investigated the possible effects of gamma-hydroxybutyric acid (GHB) on human psychomotor performance and subjective feelings important for the safety of skilled performance. METHODS: Twelve healthy volunteers, six males and six females, aged 22-36 years, participated as subjects. Drugs and placebo were administered according to a single-dose, double-blind, balanced, four-way, crossover design. Treatments were separated by a wash-out period of 1 week and consisted of placebo, lorazepam 0.03 mg x kg(-1), GHB 12.5 mg x kg(-1) and GHB 25 mg x kg(-1). Subjects' psychomotor performance was assessed at baseline and at 15, 60, 120 and 180 min after treatment. Mood was assessed using 16 visual analogue scales, before treatment and 120 min later. Psychomotor performance was measured using the following tests: Critical Flicker Fusion. Response Competition Test, Critical Tracking Task, Choice Reaction Time and Visual Vigilance Task. RESULTS: GHB at both doses had no effects on attention, vigilance, alertness, short-term memory or psychomotor co-ordination (delta-placebo, P > 0.05); calmness increased with the lower dose and contentedness decreased significantly at both doses (delta-baseline, P < 0.05); adverse effects were limited to slight subjective feelings of dizziness and dullness, which disappeared 30-60 min after administration of the dose. Lorazepam caused impairment of psychometric functions. CONCLUSION: After single therapeutic doses, GHB does not induce changes in psychomotor performance and therefore the drug does not influence the ability to drive or work. However, repeated reports of the abuse potential of GHB and its usefulness in treating ethyl alcohol addiction indicate that it may play an "agonist-like" role, which means that it should only used under close medical supervision.

Prolongation of the QT interval and the sudden infant death syndrome.

BACKGROUND: The sudden infant death syndrome (SIDS) is multifactorial in origin, but its causes remain unknown. We previously proposed that prolongation of the QT interval on the electrocardiogram, possibly resulting from a developmental abnormality in cardiac sympathetic innervation, may increase the risk of life-threatening ventricular arrhythmias and contribute to this devastating disorder. We prospectively tested this hypothesis. METHODS: Between 1976 and 1994, we recorded electrocardiograms on the third or fourth day of life in 34,442 newborns and followed them prospectively for one year. The QT interval was analyzed with and without correction for the heart rate. RESULTS: One-year follow-up data were available for 33,034 of the infants. There were 34 deaths, of which 24 were due to SIDS. The infants who died of SIDS had a longer corrected QT interval (QTc) than did the survivors (mean [+/-SD], 435+/-45 vs. 400+/-20 msec, P<0.01) and the infants who died from causes other than SIDS (393+/-24 msec, P<0.05). Moreover, 12 of the 24 SIDS victims but none of the other infants had a prolonged QTc (defined as a QTc greater than 440 msec). When the absolute QT interval was determined for similar cardiac-cycle lengths, it was found that 12 of the 24 infants who died of SIDS had a QT value exceeding the 97.5th percentile for the study group as a whole. The odds ratio for SIDS in infants with a prolonged QTc was 41.3 (95 percent confidence interval, 17.3 to 98.4). CONCLUSIONS: Prolongation of the QT interval in the first week of life is strongly associated with SIDS. Neonatal electrocardiographic screening may permit the early identification of a substantial percentage of infants at risk for SIDS, and the institution of preventive measures may therefore be possible.