Manousos E. Kambouris

Rapid extraction of fungal DNA from clinical samples for PCR amplification

A hexadecyltrimethylammonium bromide (CTAB) method for isolating fungal DNA from clinical samples, suitable for PCR amplification is described. Yeast and filamentous fungi DNA from clinical samples was amplified with primers complementary to the genes coding for rRNA, amplifying a 105 bp fragment and internal transcribed spacer primers amplifying fragments between 242 and 622 bp. The level of sensitivity was 10 +/- 5 yeast and 28 Aspergillus fumigatus CFU ml-1 of biological fluid.

Polyphasic Identification and Susceptibility to Seven Antifungals of 102 Aspergillus Isolates Recovered from Immunocompromised Hosts in Greece

ABSTRACT In this study, the first such study in Greece, we used polyphasic identification combined with antifungal susceptibility study to analyze Aspergillus clinical isolates comprising 102 common and rare members of sections Fumigati, Flavi, Terrei, Nidulantes, Nigri, Circumdati, Versicolores, and Usti. High amphotericin B MICs (>2 μg/ml) were found for 17.6% of strains. Itraconazole, posaconazole, and voriconazole MICs of >4 μg/ml were shown in 1%, 5%, and 0% of the isolates, respectively. Anidulafungin, micafungin, and caspofungin minimum effective concentrations (MECs) of ≥2 μg/ml were correspondingly recorded for 4%, 9%, and 33%, respectively, of the strains.

Recent Advances in Genetic Predisposition of Myasthenia Gravis

Myasthenia gravis (MG) is an autoimmune disease mediated by the presence of autoantibodies that bind to components of the neuromuscular junction, causing the symptoms of muscular weakness and fatigability. Like most autoimmune disorders, MG is a multifactorial, noninherited disease, though with an established genetic constituent. The heterogeneity observed in MG perplexes genetic analysis even more, as it occurs in various levels, including diverse autoantigens, thymus histopathology, and age at onset. In this context of distinct subgroups, a plethora of association studies, discussed in this review, have assessed the involvement of various HLA and non-HLA related loci in MG susceptibility, over the past five years. As expected, certain HLA alleles were strongly associated with MG. Many of the non-HLA genes, such as PTPN22 and CTLA-4, have been previously studied in MG and other autoimmune diseases and their association with MG has been reevaluated in more cohesive groups of patients. Moreover, novel risk or protective loci have been revealed, as in the case of TNIP1 and FOXP3. Although the majority of these results have been derived from candidate gene studies, the focal point of all recent genetic studies is the first genome-wide association study (GWAS) conducted on early-onset MG patients.

Test Pricing and Reimbursement in Genomic Medicine: Towards a General Strategy.

This paper aims to provide an overview of the rationale and basic principles guiding the governance of genomic testing services, to clarify their objectives, and allocate and define responsibilities among stakeholders in a health-care system, with a special focus on the EU countries. Particular attention is paid to issues pertaining to pricing and reimbursement policies, the availability of essential genomic tests which differs between various countries owing to differences in disease prevalence and public health relevance, the prescribing and use of genomic testing services according to existing or new guidelines, budgetary and fiscal control, the balance between price and access to innovative testing, monitoring and evaluation for cost-effectiveness and safety, and the development of research capacity. We conclude that addressing the specific items put forward in this article will help to create a robust policy in relation to pricing and reimbursement in genomic medicine. This will contribute to an effective and sustainable health-care system and will prove beneficial to the economy at large.

Mobile Stand-off and Stand-in Surveillance Against Biowarfare and Bioterrorism Agents

Engineered microorganisms, microorganisms traveling through massive human transportation systems to intercontinental distances and the natural processes for fast-track microorganism evolution, especially to counter antibiotics, secure the continuous presence of infectious diseases within the foreseeable future. In consequence, bioweaponeers, especially bioterrorists, will find excellent grounds for nefarious improvisations by exploiting novel agents with enhanced virulence characteristics, deliverable by various means but especially by spraying aerosolized forms. To counter the spread of such agents, along with the just as hazardous prospect of unintentional harmful agent release due to biotechnological accidents, more stringent biosurveillance schemes must be enacted, possibly 24/7, preferably integrating networking principles, state-of-the-art assets for both sensing and sampling applications and the use of inexpensive unmanned platforms, preferably mobile and even better moving in three dimensions (UAVs) so as to increase the reach, depth and persistence within surveyed space. The new tendency in post-sampling sensors will be prepackaged point-of-care assays with limited or no need of energy source and consumables, detecting 3-D structures or the nucleic acid signal for identifying agents so as to implement reactive, targeted countermeasures (decontamination, treatment). Proactive, general countermeasures, such as alert, sampling procedures and protective measures will depend on pre-sampling sensors, operating on spectroscopic principles and usually using UV-Laser Induced Fluorescence principle and carried onto manned and unmanned aerial and ground platforms designed for reconnaissance and surveillance with exchangeable payloads.

Wireless electrostimulation: a new approach in combating infection?

Electrostimulation (ES), hitherto successfully employed in wound treatment, has shown potential in antimicrobial applications, suggesting its use as synergistic to or replacement of antibiotics. The differential susceptibility of pathogens and host tissue and organs to various ES modalities might allow selective use against specific infections. The use of ES is cheaper in terms of development/testing, routine application and environmental footprint. If extensive substitution of chemical compounds is achieved, the development of resistance might be reversed through negative selection. A promising setup of ES seems to be the noncontact current transfer, due to low amperage similar to innate bioelectricity, painlessness, simple logistics and low risk for treatment-caused infection.

Wireless Micro Current Stimulation technology improves firework burn healing: Clinical applications of WMCS technology

Fireworks are used worldwide during national and cultural celebrations, but they often cause moderate to severe injuries. A young male was injured by fireworks during festivities, as a unit hit him at the right arm causing severe and extended (second degree) burn. The patient volunteered for treatment with the Wetling-W200 Wireless Micro Current Stimulation (WMCS) device, an innovative, noninvasive technology to transfer current wirelessly to wound site. After 10 sessions of 60 minutes, with the spraying intensity set at 1.5 microamperes, the patient had completely recovered in just 10 days post initial treatment; pain had been greatly reduced after the first two sessions and continued to decline after every session. The treatment sessions had been unobtrusive and painless and there was no infection or other complication, despite the fact that no painkiller or antibiotic drugs had been administered. WMCS technology seems an effective therapeutic option for firework skin burns.