Effects of cocaine and heroin, and their combination, on the development rate of Calliphora vomitoria (Diptera: Calliphoridae).

Insects present on or near decomposing bodies are collected by forensic entomologists and used to estimate the post-mortem interval. Drugs metabolized by a person before death may affect the rate of development of insects feeding on the corpse. This study aimed to determine the effects of cocaine and heroin main metabolites on the development rate of the Calliphora vomitoria (Diptera: Calliphoridae) and their implications on minimum post-mortem interval determination. Groups of 250 eggs each were placed into four separate pots of 150 g of minced pork meat being either un-spiked, or spiked with benzoylecgonine, morphine, or a combination of both. Larval length (mm) and weight (mg) measurements were taken twice daily and the rate of development of the insects' life cycle was monitored until eclosion. Results show that cocaine-fed larvae developed less in length and weight than the control group. Heroin-fed larvae showed a more fluctuating pattern, being smaller and lighter than the control group for most of their larval cycle, but overtaking them in both parameters towards pupation. Combination-fed larvae seemed to favour the effects of cocaine. The three conditions also had a significant impact on the length of the insects' life cycle. Cocaine and drug combination treatments increased the length of the second and third instar stages, but led to the shortening of pupation and accelerated eclosion. Conversely, heroin treatment led to lengthier pupation. Interestingly, the effects of the drug combination seemed to mirror more precisely those of cocaine. These findings indicate that both cocaine and heroin, singularly and in combination, have sizable effects on blowflies' development rates, potentially biasing post-mortem interval estimations.

Validation of presumptive tests for non-human blood using Kastle Meyer and Hemastix reagents.

Kastle Meyer and Hemastix reagents are presumptive tests commonly used in forensic casework for the detection of blood, and their suitability has been reviewed in numerous publications. However, studies to date have focused on the validation of these tests on human blood alone, and no published work has looked at the sensitivity, specificity and effect on DNA analysis when using these reagents to presumptively test for animal blood. The aim of this study was to validate the two reagents for use with animal blood, and compare their performance in order to choose the best test based on the circumstances in wildlife crime investigation. The sensitivity, specificity, stability and robustness of the methods were assessed by experiments with dilutions of animal blood (from 1:4 to 1:65536) using direct and indirect (rub) tests, potential interfering substances, blood sources from different species and aged blood. The effects of the two reagents on subsequent DNA analysis were also investigated. During the direct tests, Kastle Meyer showed a higher sensitivity, detecting blood down to a dilution of 1:16,384, one order of magnitude lower than Hemastix. However during the rub test, Hemastix showed a higher sensitivity, detecting blood down to a dilution of 1:64 on porous materials while Kastle Meyer was positive only down to a dilution of 1:16. Moreover, when using the same swab for presumptive testing and DNA extraction, Hemastix testing allowed amplification of a sufficient amount of DNA for species identification at its limit of sensitivity on porous materials (1:64) while Kastle Meyer inhibited most amplification of DNA at its less sensitive limit of 1:16 dilution. On the other hand, Hemastix showed a much lower specificity, producing false positive results when exposed to tomato, potato, rust, avian uric acid, bleach and sink rot, while Kastle Meyer only produced a faint positive reaction from potato. Both tests performed equally well detecting fresh blood of different animal species. The stability test gave comparable results among the tests except for aged fish blood stains, where the Kastle Meyer test performed poorly. Owing to its ease of use, higher sensitivity, and lack of interference with downstream DNA analysis, and despite its reduced specificity compared to Kastle Meyer, the Hemastix method is more appropriate for use in wildlife crime investigations. Positive results would always be confirmed with DNA analysis and the low interference of the reagent will allow the use of a single swab for presumptive testing and DNA sampling.

Preliminary results of a new approach for three-dimensional reconstruction of Dynamic AngioThermography (DATG) images based on the inversion of heat equation.

Dynamic AngioThermography (DATG) is a contact-plate technique capable of producing a digital representation of breast vascularity. The inception and growth of a tumor are associated with neoangenesis, which may result in a demonstrable alteration in the regional blood flow, while in normal health conditions the vascularity remains unchanged throughout life. DATG, if included in the clinical evaluation for breast cancer, could potentially improve the accuracy of the diagnosis of this disease. Conventional DATG is limited, however, in that it is a projection (i.e. two-dimensional) imaging technique that does not provide any information on the depth and its effect on the pattern of the perfusion revealed by this technique. In fact, the blood pattern is detected by projecting temperature signals on the plate, thus acquiring a digital two-dimensional image. In this article we propose a new approach for extracting information on depth through the inversion of the Fourier heat equation. The idea is to extract the information along the third axis while acquiring and analyzing the temporal sequence during the process of image formation. The method implemented has been tested on a dedicated "electric phantom" and in one in vivo experiment. In spite of the limits of these preliminary tests, the experimental results have shown that this method makes it possible to obtain a 3D representation of the vascularity. Although it appears to be promising, further validation and characterization of our technique are required.

Dental status and 3D reconstruction of the malocclusion of the famous singer Farinelli (1705-1782).

The famous castrato singer Carlo Broschi, better known as Farinelli (1705-1782), was exhumed by our research group in July 2006 to study his skeletal remains and reconstruct his osteobiography. He was castrated before puberty to preserve his high voice into adulthood. The osteological study has revealed several skeletal features probably related to the effects of castration (Hyperostosis frontalis interna, long limb bones, persistence of epiphyseal lines, osteoporosis) (Belcastro et al., 2011). Here we present the study of the teeth and maxilla-mandibular region using classic and tomographic morphological methods. Considering the subject's age and the period during which he lived, his oral health conditions were good. On the basis of the very pronounced anomalous vestibular buccal wear, a overbite visualized by 3D reconstruction, was hypothesized. This facial disharmony is of particular interest when considering Farinelli's extraordinary singing qualities and stage presence.

Critical steps in fluoroquinolones and carbapenems prescriptions: results from a prospective clinical audit.

Antibiotic misuse is associated with emergence of resistance and high expenditures. Fluoroquinolones (FQ) and carbapenems (CP) are drugs with considerable potential of resistance development and its disseminated use is a concern. We undertook a prospective clinical audit to evaluate prescriptions of FQ and CP in a multistep process. Each prescription was unfolded in the following steps: indication for antimicrobial therapy; adequacy of initial prescription, dosage and route; previous cultures; and parenteral-oral transition. There was no antibiotics indication in 8.9% of FQ and 1.5% of CP group (p = 0.07). In CP 25.8% of initial schemes were inappropriate (21% in FQ). Lack of switch to oral therapy comprised 25% of monthly costs of FQ. Inadequacy in initial choice accounted for 13.6% of CP expenses. We concluded that, in spite of infection control restrictive policies, inappropriateness of antibiotic usage is worrisome. Clinical audit in a multistep approach may identify possible flaws in this process.

High-resolution 3D scaffold model for engineered tissue fabrication using a rapid prototyping technique.

Rapid prototyping, automatic image processing (computer-aided design (CAD)) and computer-aided manufacturing techniques are opening new and interesting prospects for medical devices and tissue engineering, especially for hard tissues such as bone. The development of a bone high-resolution scaffold prototype using these techniques is described. The results testify to the fidelity existing between microtomographic reconstruction and CAD. Furthermore, stereolithographic manufacturing of this scaffold, which possesses a high degree of similarity to the starting model as monitored by morphological evaluations (mean diameter 569 +/- 147 microm), represents a promising result for regenerative medicine applications.

Digital radiography using an EBCCD-based imaging device

In the last few years, large efforts were devoted to the development of new intensified imaging systems to be applied mainly in medical radiographic devices for the reduction of the dose irradiated to the patient but also in high-energy physics for particle tracking or in astronomy for low-light imaging. Among a number of different image-intensifier systems, electron-bombarded charge coupled devices (EBCCD) present interesting characteristics providing the opportunity to be used also for special application such as X-ray imaging in industrial field. For this purpose, a prototype EBCCD image-intensifier tube suitable for X-ray digital radiography (DR) and computed tomography (CT) has been set up by the Physics Department of the University of Bologna (Italy) and Geosphaera Research Center of Moscow (Russia). Results concerning tests of EBCCD device and DR analysis of industrial components are presented and discussed.

Seroprevalence and anti-HEV persistence in the general population of the Republic of San Marino.

The prevalence of anti-HEV was assessed in 2,233 subjects aged 20-79 years in the Republic of San Marino in the years 1990-1991. The sera were tested by ELISA and further confirmed by Western blot (WB) analysis. The overall anti-HEV prevalence was 1.5%. A significant trend by age was observed. Anti-HEV prevalence was 0.6% in subjects <30 years and 3.3% in those older than 70 years of age. Family size larger than four persons (OR = 3.8; 95% CI = 1.8-13.2) was the sole independent predictor of anti-HEV positivity in the multivariate analysis. Anti-HAV and anti-HEV prevalences did not show a parallel trend by age. No association was found either between hepatitis E virus (HEV) or hepatitis C virus (HCV) infections. Follow-up samples 5 years apart were available for 38 out of 54 (70%) anti-HEV ELISA-positive subjects. Eight out of 22 (37%) WB-confirmed anti-HEV-positive subjects were still anti-HEV-positive after 5 years. However, anti-HEV remained positive in all but two (75%) of the subjects with WB-confirmed ELISA positivity value of S/CO > or = 2 (cutoff 1.2), but in only 2 out of the 14 subjects (14%) with a WB-confirmed ELISA positivity value of S/CO < 2 (P < 0.005). None of the 16 subjects ELISA-positive but not WB-confirmed was anti-HEV-positive 5 years apart. Therefore, only a relative proportion of subjects once infected with HEV maintain for at least 5 years anti-HEV antibodies.

Fulminant hepatitis associated with hepatitis A virus superinfection in patients with chronic hepatitis C.

BACKGROUND: Hepatitis A virus (HAV) infection rarely causes fulminant hepatic failure in people with no underlying liver disease. There are limited data on the course of this infection in patients with chronic hepatitis B and chronic hepatitis C. METHODS: We prospectively followed, from June 1990 to July 1997, 595 adults with biochemical and histologic evidence of chronic hepatitis B (163 patients) or chronic hepatitis C (432 patients) who were seronegative for HAV antibodies. All were tested every four months for serum IgM and IgG antibodies to HAV. RESULTS: Twenty-seven patients acquired HAV superinfection, 10 of whom had chronic hepatitis B and 17 of whom had chronic hepatitis C. One of the patients with chronic hepatitis B, who also had cirrhosis, had marked cholestasis (peak serum bilirubin level, 28 mg per deciliter [479 micromol per liter]); the other nine had uncomplicated courses of hepatitis A. Fulminant hepatic failure developed in seven of the patients with chronic hepatitis C, all but one of whom died. The other 10 patients with chronic hepatitis C had uncomplicated courses of hepatitis A. CONCLUSIONS: Although most patients with chronic hepatitis B who acquired HAV infection had an uncomplicated course, patients with chronic hepatitis C had a substantial risk of fulminant hepatitis and death associated with HAV superinfection. Our data suggest that patients with chronic hepatitis C should be vaccinated against hepatitis A.